TY - BOOK T1 - Future Strategies for Tissue and Organ Replacement A1 - Polak JM A1 - Hench LL A1 - Kemp P Y1 - 2002/// SN - 1-8609-4310-1 N2 - - ER - TY - CHAP T1 - Application of MRI to Cell Tracking A1 - Bhakoo, K A1 - Chapon, C A1 - Jackson, J A1 - Jones, W ED - G.A. Webb T2 - Handbook of Modern Magnetic Resonance: Biological Sciences Y1 - 2006/// PB - Springer SP - (in press) N2 - - ER - TY - CHAP T1 - Application of MRI to Stem Cell Imaging A1 - Bhakoo, K A1 - Jones, W A1 - Jackson, J A1 - Chapon, C ED - N.A. Habib, M.Y. Gordon, N. Levicar et al T2 - Stem Cell Repair and Regeneration Y1 - 2005/// PB - Imperial College Press CY - London N2 - - ER - TY - CHAP T1 - Nuclear magnetic resonance spectroscopy of in the study of human liver. A1 - Lim AK A1 - Khan SA A1 - Cox IJ A1 - Taylor-Robinson SD ED - Tosi R, Tugnoli V T2 - Nuclear Magnetic Resonance Spectroscopy in the Study of Neoplastic Tissue. Y1 - 2005/// VL - 1 PB - Nova Science CY - Hauppauge, New York SN - 1-59454-258-9 SP - 295 EP - 311 N2 - - ER - TY - CHAP T1 - Pharmacological characterisation of embryonic stem cell-derived cardiomyocyte cultures A1 - Ali, N N A1 - Brito-Martins, M A1 - Gorelik, J A1 - Xu, X A1 - Korchev, Y A1 - Zhu, H A1 - Poole-Wilson, P A A1 - Fuller, S J A1 - Harding, S E ED - Habib, N., Gordon, M.Y., Levicar, N. and Jiao, L. T2 - Stem Cells:Repair and Regeneration Y1 - 2005/// PB - Imperial College Press CY - London SP - 139 EP - 147 N2 - - ER - TY - CHAP T1 - Gene Targeting in sheep. A1 - Clark AJ A1 - Cui W ED - J McWhir and A Thomson T2 - Gene targeting & embryonic stem cells (Advanced methods~) Y1 - 2004/07/01/ PB - Taylor & Francis SN - 1859963609 SP - 45 EP - 69 N2 - - ER - TY - CHAP T1 - The Application of Magnetic Resonance Imaging and Spectroscopy to Gene Therapy A1 - Bhakoo, K K A1 - Bell, J D A1 - Cox, I J A1 - Taylor-Robinson, S D T2 - Methods in Enzymology Y1 - 2004/// VL - Imaging in Biological Research M2 - 386 PB - Elsevier Inc. SP - 303 EP - 313 N2 - - UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&dopt=Abstract&list_uids=15120258&query_hl=7&itool=pubmed_DocSum ER - TY - CHAP T1 - Hormones and the gastrointestinal tract. A1 - Hammond PJ A1 - Bloom SR A1 - Bishop AE A1 - Polak JM ED - D.A. Warrell, T.M. Cox, J.D. Firth, E.J. Benz T2 - Oxford Textbook of Medicine Y1 - 2003/// M2 - 4th PB - Oxford University Press CY - Oxford, U.K. SN - 0-19-262922-0 SP - 569 EP - 576 N2 - - ER - TY - CHAP T1 - Pathology of pulmonary hypertension A1 - Bishop AE A1 - Polak JM ED - Banner N R, Polak J M, Yacoub M T2 - Lung Transplantation Y1 - 2003/// PB - Cambridge University Press CY - Cambridge SP - 19 EP - 28 N2 - - ER - TY - CHAP T1 - Pathology pf Pulmonary Hypertension A1 - Bishop AE A1 - Polak JM ED - N. Banner, M.H. Yacoub T2 - Lung Transplantation Y1 - 2003/// PB - Cambridge University Press CY - Cambridge, U.K. SP - 20 EP - 28 N2 - - ER - TY - CHAP T1 - The anatomy, organization and ultrastructure of the Islets of Langerhans A1 - Bishop AE A1 - Polak JM ED - J. Pickup and G. Williams T2 - Textbook of Diabetes Y1 - 2002/// PB - Blackwell Scientific Ltd CY - Oxford, U.K. SN - 0-6320-5915-X SP - 10.1 EP - 10.6 N2 - - ER - TY - CHAP T1 - The anatomy, organization and ultrastructure of the Islets of Langerhans A1 - Bishop AE A1 - Polak JM T2 - Textbook of Diabetes Y1 - 2002/// SN - 0-6320-5915-X SP - 10 EP - 10 N2 - - ER - TY - CHAP T1 - NCX overexpression in cardiac myocytes A1 - Terracciano CMN ED - J. Lytton; P. Schnetkamp, L. Hryshko, M. Blaunstein T2 - Cellular and molecular physiology of Sodium - Calcium exchange Y1 - 2002/// PB - New York Academy of Science CY - New York SP - 520 EP - 527 N2 - - ER - TY - CHAP T1 - Stem Cells: sources and applications A1 - Buttery LDK A1 - Polak JM T2 - Future Strategies for Tissue and Organ Replacement Y1 - 2002/// SN - 1-8609-4310-1 SP - 351 EP - 368 N2 - - ER - TY - CHAP T1 - Nitric oxide and other vasoactive agents A1 - Buttery LDK A1 - Mancini L A1 - Moradi-Bidhendi N A1 - O'Shaughnessy MC A1 - Polak JM A1 - MacIntyre I T2 - Principles of Bone Biology Y1 - 2002/// SN - 0-1209-8652-3 SP - 995 EP - 1013 N2 - - ER - TY - CHAP T1 - The gut and the autonomic nervous system A1 - Bishop AE A1 - Polak JM ED - Sir Roger Bannister, C.J. Mathias T2 - Autonomic Failure Y1 - 2001/// PB - Oxford University Press CY - Oxford, U.K. SN - 0-1926-2850-X SP - 117 EP - 125 N2 - - ER - TY - CONF T1 - Mouse embryonic stem cell engrafment in healthy and injured mouse lung A1 - Lane, S A1 - Rippon, H A1 - Takata, M A1 - Mahadeva, R A1 - Bishop, AE U1 - Conference of the Tissue-Engineering-and-Regenerative-Medicine-International-Society (TERMIS-EU) Y1 - 2007/07// Y2 - // VL - 13 SP - 1731 EP - 1731 N2 - - ER - TY - CONF T1 - Embryonic stem cells for the engineering and regeneration of mineralized tissues A1 - Buttery, LDK A1 - Polak, JM U1 - Conference of the NATO-Advanced-Study-Institute on Learning from Nature How to Design New Implantable Biomaterials Y1 - 2004/// Y2 - // VL - 171 SP - 199 EP - 204 N2 - - ER - TY - CONF T1 - Stem cells and bioactive materials A1 - Bielby, RC A1 - Polak, JM U1 - Conference of the NATO-Advanced-Study-Institute on Learning from Nature How to Design New Implantable Biomaterials Y1 - 2004/// Y2 - // VL - 171 SP - 181 EP - 198 N2 - - ER - TY - CONF T1 - Endothelial nitric oxide synthase is an important regulator of osteoblast growth and differentiation. A1 - Afzal, F A1 - Nohadani, M A1 - Huang, PL A1 - Polak, JM A1 - Buttery, LDK U1 - 24th Annual Meeting of the American-Society-for-Bone-and-Mineral-Research Y1 - 2002/09// Y2 - // VL - 17 SP - S149 EP - S149 N2 - - ER - TY - CONF T1 - A genetic basis for biomedical materials A1 - Hench, LL A1 - Xynos, ID A1 - Edgar, AJ A1 - Buttery, LDK A1 - Polak, JM U1 - Conference on Materials Science and Engineering Y1 - 2002/// Y2 - // SP - 283 EP - 296 N2 - - ER - TY - CONF T1 - Materials in medicine A1 - Bielby, RC A1 - Buttery, LDK A1 - Polak, JM U1 - Conference on Materials Science and Engineering Y1 - 2002/// Y2 - // SP - 135 EP - 151 N2 - - ER - TY - CONF T1 - Study of osteoblast differentiation and proliferation on the surface of binary bioactive gel-glasses A1 - Bielby, RC A1 - Saravanapavan, P A1 - Polak, JM A1 - Hench, LL U1 - 14th International Symposium on Ceramics in Medicine (BIOCERAMICS-14) Y1 - 2002/// Y2 - // VL - 218-2 SP - 269 EP - 272 N2 - - ER - TY - CONF T1 - Pelvic nerve plexus trauma at radical hysterectomy and simple hysterectomy - The nerve content of the uterine supporting ligaments A1 - Butler-Manuel, SA A1 - Buttery, LDK A1 - A'Hern, RP A1 - Polak, JM A1 - Barton, DPJ U1 - 46th Annual Meeting of the Society-for-Gynecologic-Investigation Y1 - 2000/08/15/ Y2 - // VL - 89 SP - 834 EP - 841 N2 - - ER - TY - JFULL T1 - The Transcriptional Corepressor RIP140 Regulates Oxidative Metabolism in Skeletal Muscle. A1 - Seth, A A1 - Steel, JH A1 - Nichol, D A1 - Pocock, V A1 - Kumaran, MK A1 - Fritah, A A1 - Mobberley, M A1 - Ryder, TA A1 - Rowlerson, A A1 - Scott, J A1 - Poutanen, M A1 - White, R A1 - Parker, M J1 - Cell Metabolism Y1 - 2007/09// VL - 6 SP - 236 EP - 245 ER - TY - JFULL T1 - The impact of chromatin modifiers on the timing of locus replication in mouse ES cells. A1 - Jorgensen, HF A1 - Azuara, V A1 - Amoils, S A1 - Spivakov, M A1 - Terry, A A1 - Nesterova, T A1 - Cobb, BS A1 - Ramsahoye, B A1 - Merkenschlager, M A1 - Fisher, AG J1 - Genome Biol Y1 - 2007/08/17/ VL - 8 SN - 1465-6914 SP - R169 EP - R169 N2 - ABSTRACT: BACKGROUND: The time of locus replication during S-phase is tightly regulated and correlates with chromatin state. Embryonic stem (ES) cells have an unusual chromatin profile where many developmental regulator genes, that are not yet expressed, are marked by both active and repressive histone modifications. This poised or bivalent state is also characterized by locus replication in early S-phase in ES cells, while replication timing is delayed in cells with restricted developmental options. RESULTS: Here we used a panel of mutant mouse ES cell lines lacking important chromatin modifiers to dissect the relationship between chromatin structure and replication timing. We show that temporal control of satellite DNA replication is sensitive to loss of a variety of chromatin modifiers including Mll, Eed, Dnmt1, Suv39h1/h2 and Dicer. The replication times of many single copy loci, including a 5 Mb contiguous region surrounding the Rex1 gene were retained in chromatin modifier mutant ES cells, although a subset of loci were affected. Conclusions; This analysis demonstrates the importance of chromatin modifiers for maintaining correct replication of satellite sequences in pluripotent ES cells and highlights the sensitivity of some single copy loci to the influence of chromatin modifiers. Abundant histone acetylation is shown to correlate well with early replication. Surprisingly, loss of DNA methylation or histone methylation was tolerated by many loci suggesting that these modifications may be less influential for the timing of euchromatin replication. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17705870&query_hl=1 ER - TY - JFULL T1 - The Nuclear Receptor Cofactor RIP140 is Required for the Regulation of Hepatic Lipid and Glucose Metabolism by LXR. A1 - Herzog, B A1 - Hallberg, M A1 - Seth, A A1 - Woods, A A1 - White, R A1 - Parker, MG J1 - Mol Endocrinol Y1 - 2007/08/07/ SN - 0888-8809 N2 - The liver X receptors (LXRs) are nuclear receptors which play important roles in the regulation of lipid metabolism. In this study, we demonstrate that RIP140 is a cofactor for LXR in liver. Analysis of RIP140 null mice and hepatocytes depleted of RIP140 indicate that the cofactor is essential for the ability of LXR to activate the expression of a set of genes required for lipogenesis. Furthermore we demonstrate that RIP140 is required for the ability of LXR to repress the expression of the PEPCK gene in Fao cells and mice. Thus, we conclude that the function of RIP140 as a cofactor for LXR in liver varies according to the target genes and metabolic process, serving as a coactivator in lipogenesis but as a corepressor in gluconeogenesis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17684114&query_hl=1 ER - TY - JFULL T1 - Functional variants of the central bile acid sensor FXR identified in intrahepatic cholestasis of pregnancy. A1 - Van Mil, SW A1 - Milona, A A1 - Dixon, PH A1 - Mullenbach, R A1 - Geenes, VL A1 - Chambers, J A1 - Shevchuk, V A1 - Moore, GE A1 - Lammert, F A1 - Glantz, AG A1 - Mattsson, LA A1 - Whittaker, J A1 - Parker, MG A1 - White, R A1 - Williamson, C J1 - Gastroenterology Y1 - 2007/08// VL - 133 SN - 0016-5085 SP - 507 EP - 516 N2 - BACKGROUND AND AIMS: Intrahepatic cholestasis of pregnancy (ICP) is characterized by liver impairment, pruritus, and elevated maternal serum bile acids. It can cause premature delivery and intrauterine death. Bile acid synthesis, metabolism, and transport are regulated by the bile acid sensor FXR, and we hypothesized that genetic variation in FXR confers susceptibility to ICP. METHODS: The coding regions and intron/exon boundaries of FXR were sequenced in 92 British ICP cases of mixed ethnicity. Subsequently, a case-control study of allele frequencies of these variants in 2 independent cohorts of Caucasian ICP patients and controls was performed. Variants were cloned into an FXR expression plasmid and tested in functional assays. RESULTS: We identified 4 novel heterozygous FXR variants (-1g>t, M1V, W80R, M173T) in ICP. W80R was not present in Caucasians and M1V was detected uniquely in 1 British case. M173T and -1g>t occur both in Caucasian cases and controls, and we found a significant association of M173T with ICP (OR, 3.2; 95% confidence interval, 1.1-11.2; P = .02) when the allele frequencies of both Caucasian cohorts were analyzed together. We demonstrate functional defects in either translation efficiency or activity for 3 of the 4 variants (-1g>t, M1V, M173T). CONCLUSIONS: This is the first report of functional variants in FXR. We propose that these variants may predispose to ICP, and because FXR has a central role in regulating bile and lipid homeostasis they may be associated with other cholestatic and dyslipidemic disorders. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17681172&query_hl=1 ER - TY - JFULL T1 - Pseudoepitheliomatous hyperplasia - an unusual reaction following tattoo: report of a case and review of the literature. A1 - Cui, W A1 - McGregor, DH A1 - Stark, SP A1 - Ulusarac, O A1 - Mathur, SC J1 - Int J Dermatol Y1 - 2007/07// VL - 46 SN - 0011-9059 SP - 743 EP - 745 N2 - A 59-year-old woman presented with an itchy and uncomfortable raised lesion at a tattoo site (Fig. 1) on the lateral aspect of the left leg, just above the ankle. The tattoo had been placed 2 years before her presentation and the tattoo site was sun exposed. Immediately after she had the tattoo, she noticed redness of the skin. After a week, a pruritic and red scaly nodule developed that continued to gradually enlarge until her presentation. The patient had tried topical vitamin A and D ointment with no relief. The patient also had tattoos on the arms without any noticeable skin changes. The patient reported that the tattoo procedure on her leg was more painful than that on her arms, and was performed by a different (and perhaps inexperienced) tattoo artist. The original tattoo contained red, green, and yellow pigments. A diagnosis of tattoo granuloma was considered; squamous cell carcinoma and fungal infection were included in the differential diagnosis. A punch biopsy was performed, followed by complete surgical excision of the lesion with a split-thickness skin graft from the right thigh. The skin excision specimen showed a 3 x 2.5-cm granular and pitted pink lesion with well-demarcated, somewhat irregular borders. The lesion was raised 0.5 cm above the skin surface. The lesion was present in the center of the original tattoo. Portions of the original tattoo with green and blue-green pigmentation were visible on either side of the lesion. No satellite lesions were identified. Microscopically, the raised lesion demonstrated striking pseudoepitheliomatous hyperplasia, with irregular acanthosis of the epidermis and follicular infundibula, hyperkeratosis, and parakeratosis (Fig. 2). Follicular plugging was present with keratin-filled cystic spaces. There was a brisk mononuclear inflammatory infiltrate in the dermis, composed primarily of lymphocytes, with admixed plasma cells and histiocytes. Giant cells were occasionally identified. Dermal pigment deposition was noted both within the lesion and in the surrounding skin, corresponding to the original tattoo. Variable dermal fibrosis was noted, with thick collagen bundles in some areas. There was no evidence of epidermal keratinocytic atypia, dyskeratosis, or increased suprabasal mitotic activity. Special stains (periodic acid-Schiff and acid-fast) for microorganisms were negative. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17614808&query_hl=1 ER - TY - JFULL T1 - Heart tissue engineering using a novel elastomer and ES-derived cardiac cells A1 - Chena, QZ A1 - Wright, JS A1 - Harding, SE A1 - Junaid, S A1 - Hansen, U A1 - Jawad, H A1 - Boccaccini, AR A1 - Ali, NN J1 - TISSUE ENG Y1 - 2007/07// VL - 13 SN - 1076-3279 SP - 1703 EP - 1703 ER - TY - JFULL T1 - Therapeutics of vein graft intimal hyperplasia: 100 years on. A1 - Wallitt, EJ A1 - Jevon, M A1 - Hornick, PI J1 - Ann Thorac Surg Y1 - 2007/07// VL - 84 SN - 1552-6259 SP - 317 EP - 323 N2 - Intimal hyperplasia is central to the pathology of vein graft re-stenosis, and despite considerable advances in our understanding of vascular biology since it was first described 100 years ago, it is still a significant clinical problem. Recent decades have seen the development of many new therapeutic agents aimed at treating this condition, but the successes of laboratory studies have not been replicated in the clinic yet. This review discusses these therapeutic agents, how their modes of action relate to the pathogenesis of vein graft intimal hyperplasia, and considerations of ways in which such therapy may be improved in the future. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17588453&query_hl=1 ER - TY - JFULL T1 - Mouse embryonic stem cell engrafment in healthy and injured mouse lung A1 - Lane, S A1 - Rippon, H A1 - Takata, M A1 - Mahadeva, R A1 - Bishop, AE J1 - TISSUE ENG Y1 - 2007/07// VL - 13 SN - 1076-3279 SP - 1731 EP - 1731 ER - TY - JFULL T1 - Stem cells in lung repair and regeneration. A1 - Lane, S A1 - Rippon, HJ A1 - Bishop, AE J1 - Regen Med Y1 - 2007/07// VL - 2 SN - 1746-076X SP - 407 EP - 415 N2 - Repair or regeneration of defective lung tissue would be of great clinical use. Potential cellular sources for the regeneration of lung tissue in vivo or lung tissue engineering in vitro include endogenous pulmonary stem cells, extrapulmonary circulating stem cells and embryonic stem cells. This review summarizes the recent research on each of these stem cell types and their potential for use in the treatment of lung injury and disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17635048&query_hl=1 ER - TY - JFULL T1 - Visfatin expression is hormonally regulated by metabolic and sex hormones in 3T3-L1 pre-adipocytes and adipocytes. A1 - MacLaren, R A1 - Cui, W A1 - Cianflone, K J1 - Diabetes Obes Metab Y1 - 2007/07// VL - 9 SN - 1462-8902 SP - 490 EP - 497 N2 - AIM: The novel adipokine visfatin has 'insulin-mimicking' effects and is increased in models of diet-induced obesity, but factors that regulate visfatin have not been fully elucidated. METHODS: In order to determine visfatin regulation in adipocyte development and metabolism, as well as in pathophysiological conditions related to metabolic syndrome, endogenous visfatin expression was measured in 3T3-L1 pre-adipocytes and adipocytes using real-time reverse transcriptase polymerase chain reaction (real-time RT-PCR). RESULTS: A marked increase in visfatin expression was observed during differentiation, with a 2.2-fold increase between preconfluent and 2-day confluent cells even before differentiation was initiated. A further 4.1-fold increase was induced from day 0 to day 9 of differentiation (overall ninefold). Overnight incubation with dexamethasone (10(-8) to 10(-2) M) increased visfatin expression in both pre-adipocytes (1.5- to 3.3-fold, p < 0.05) and adipocytes (1.9-fold, p < 0.01). All other treatments decreased visfatin expression. In pre-adipocytes, visfatin expression decreased by 23% at a concentration of 1 microM insulin, 15% at 1-15 nM T3, 15% at 10 nM-1 microM progesterone, 33-44% at 10 nM-1 microM testosterone, 50% with palmitate and 30% with oleate (p < 0.05 for all). In adipocytes, insulin had a much greater effect, decreasing visfatin by 77% at 100 nM (p < 0.01), whereas oleate and sex hormones did not affect visfatin expression. However, tumor necrosis factor alpha, which had no effect on pre-adipocytes, significantly decreased visfatin in adipocytes by 26% at 10 ng/ml (p < 0.05). Interestingly, the thiazolidinedione (TZD) rosiglitizone also decreased visfatin by 28% at a concentration of 1 microM (p < 0.01). CONCLUSION: In summary, while the mechanism of visfatin action remains to be elucidated, the clear effects of multiple hormones on visfatin expression support a physiological role. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17587391&query_hl=1 ER - TY - JFULL T1 - Receptor interacting protein 140 regulates expression of uncoupling protein 1 in adipocytes through specific peroxisome proliferator activated receptor isoforms and estrogen-related receptor alpha A1 - Debevec , D A1 - Christian , M A1 - Morganstein , D A1 - Seth , A A1 - Herzog , B A1 - Parker, M A1 - White, R J1 - Molecular Endocrinology Y1 - 2007/07// VL - 21 SP - 1581 EP - 1592 ER - TY - JFULL T1 - Direct effects of apelin on cardiomyocyte contractility and electrophysiology. A1 - Farkasfalvi, K A1 - Stagg, MA A1 - Coppen, SR A1 - Siedlecka, U A1 - Lee, J A1 - Soppa, GK A1 - Marczin, N A1 - Szokodi, I A1 - Yacoub, MH A1 - Terracciano, CM J1 - Biochem Biophys Res Commun Y1 - 2007/06/15/ VL - 357 SN - 0006-291X SP - 889 EP - 895 N2 - Apelin, the ligand for the angiotensin receptor like-1, has been implicated in the pathogenesis of atrial fibrillation and heart failure. However, it is unknown if apelin has direct effects on cardiomyocyte contractility and electrophysiology. APJ-like immunoreactivity was localized to T-tubules and intercalated disc area in isolated adult rat ventricular myocytes. Apelin (1 nM) significantly increased sarcomere shortening in normal as well as failing cardiomyocytes. The transient increase in shortening was not accompanied by increased [Ca(2+)] transient amplitude. Apelin significantly activated the sarcolemmal Na(+)/H(+) exchanger (NHE) and increased intracellular pH. Moreover, apelin (10 nM) increased conduction velocity in monolayers of cultured neonatal rat cardiac myocytes. Our results demonstrate for the first time that apelin has direct effects on the propagation of action potential and contractility in cardiomyocytes. One of the mechanisms involved in the inotropic effect may be an increased myofilament sensitivity to Ca(2+) as apelin enhanced the activity of NHE with consequent intracellular alkalinization. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17466269&query_hl=1 ER - TY - JFULL T1 - IDENTIFICATION AND CHARACTERIZATION OF A DYSFUNCTIONAL CARDIAC MYOCYTE PHENOTYPE: ROLE OF BACTERIA, TOLL-LIKE RECEPTORS, AND ENDOTHELIN. A1 - Patel, TA A1 - Harding, SE A1 - Belcher, E A1 - Warner, TD A1 - Mitchell, JA J1 - Shock Y1 - 2007/06/07/ SN - 1073-2322 N2 - Cardiac myocyte dysfunction is clearly identified as underlying the acute heart failure associated with bacterial infection, as well as the chronic syndrome following cardiac damage, but the mechanisms leading to dysfunction in each case are not fully established. It is thought that local hormones such as endothelin 1 (ET-1) can increase the risk of heart failure in acute or chronic conditions. In the current study, we characterize myocytes as populations and identify a novel phenotype of the ventricular cardiac myocyte that does not contract appropriately on electrical stimulation. The noncontractile cardiac myocytes were viable and had normal calcium transients. The proportion of noncontractile cardiac myocytes was increased by bacteria (gram-positive Staphylococcus aureus or gram-negative Escherichia coli). Using selective ligands or myocytes from genetically modified mice, we established that the effects of S. aureus were mediated by Toll-like receptor 2/6 and of E. coli by Toll-like receptor 4. The transition to the noncontractile phenotype was strongly inhibited by ETA antagonism but unaffected by inhibition of NOS, suggesting that ET-1 and not NO mediates this phenomenon. These results are the first to describe the characteristics of this noncontractile phenotype and the mechanisms of its induction by bacteria. Description of the myocyte population, instead of effects only on individual cells, will be more relevant to the prediction of the depression of cardiac function. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17558348&query_hl=1 ER - TY - JFULL T1 - Reticulocyte hemoglobin content in the diagnosis of iron deficiency in Chinese pre-menopausal women. A1 - Luo, D A1 - Chen, Y A1 - Wu, W A1 - Zhang, F A1 - Xu, J A1 - Cui, W A1 - Li, SL A1 - Li, RS J1 - Chin Med J (Engl) Y1 - 2007/06/05/ VL - 120 SN - 0366-6999 SP - 1010 EP - 1012 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17624271&query_hl=1 ER - TY - JFULL T1 - Optimizing leaf width for multileaf collimator A1 - Cui, W A1 - Dai, J J1 - MED PHYS Y1 - 2007/06// VL - 34 SN - 0094-2405 SP - 2478 EP - 2478 ER - TY - JFULL T1 - Clenbuterol affects rat ventricular myocyie contractility via an inhibitory G protein-mediated pathway A1 - Siedlecka, U A1 - Arora, M A1 - Soppa, GKR A1 - Lee, J A1 - Stagg, MA A1 - Harding, SE A1 - Yacoub, MH A1 - Terracciano, CMN J1 - J MOL CELL CARDIOL Y1 - 2007/06// VL - 42 SN - 0022-2828 SP - S49 EP - S49 ER - TY - JFULL T1 - Endogenous Flt3-ligand levels are not altered in mice after a severe burn and infection A1 - Bohannon, J A1 - Cui, W A1 - Toliver-Kinsky, T J1 - SHOCK Y1 - 2007/06// VL - 27 SN - 1073-2322 SP - 48 EP - 48 ER - TY - JFULL T1 - Neutralization of plasmacytoid dendritic cells decreases the Flt3 ligand-induced enhancement of immune cell activation in response to a burn wound infection A1 - Toliver-Kinsky, T A1 - Cui, W A1 - Sherwood, E J1 - SHOCK Y1 - 2007/06// VL - 27 SN - 1073-2322 SP - 48 EP - 48 ER - TY - JFULL T1 - Phosphodiesterase activity in isolated cardiomyocytes from rat, guinea pig and human heart A1 - Johnson, WB A1 - Katugampola, S A1 - Napier, C A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2007/06// VL - 42 SN - 0022-2828 SP - S116 EP - S116 ER - TY - JFULL T1 - Which gene, Reg2 or Reg3beta, was targeted that affected liver regeneration? A1 - Liu, JL A1 - Cui, W J1 - Hepatology Y1 - 2007/06// VL - 45 SN - 0270-9139 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17538937&query_hl=1 ER - TY - JFULL T1 - A novel Z-groove index characterizing myocardial surface structure and its modification in failing human heart A1 - Gorelik, J A1 - Yang, L A1 - Zhang, Y A1 - Lab, M A1 - Korcbev, YE A1 - del Monte, F A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2007/06// VL - 42 SN - 0022-2828 SP - S153 EP - S153 ER - TY - JFULL T1 - Differential effects of SERCA2a transfection on ventricular arrhythmia susceptibility in the normal rat heart A1 - Lyon, AR A1 - Poole-Wilson, PA A1 - Peters, NS A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2007/06// VL - 42 SN - 0022-2828 SP - S20 EP - S20 ER - TY - JFULL T1 - MRI tracking of systemically administered bone marrow stem cells A1 - Carr, CA A1 - Stuckey, DJ A1 - Tatton, L A1 - Tyler, DJ A1 - Harding, SE A1 - Clarke, K J1 - J MOL CELL CARDIOL Y1 - 2007/06// VL - 42 SN - 0022-2828 SP - S88 EP - S89 ER - TY - JFULL T1 - Responses to endothelin in human embryonic stem cell-derived cardiomyocytes A1 - Wright, J A1 - Ahmet, S A1 - Harding, SE A1 - Ali, NN J1 - J MOL CELL CARDIOL Y1 - 2007/06// VL - 42 SN - 0022-2828 SP - S88 EP - S88 ER - TY - JFULL T1 - Direct intramyocardial but not intracoronary injection of bone marrow cells induces ventricular arrhythmias in a rat chronic ischemic heart failure model. A1 - Fukushima, S A1 - Varela-Carver, A A1 - Coppen, SR A1 - Yamahara, K A1 - Felkin, LE A1 - Lee, J A1 - Barton, PJ A1 - Terracciano, CM A1 - Yacoub, MH A1 - Suzuki, K J1 - Circulation Y1 - 2007/05/01/ VL - 115 SN - 1524-4539 SP - 2254 EP - 2261 N2 - BACKGROUND: Therapeutic efficacy of bone marrow (BM) cell injection for treating ischemic chronic heart failure has not been established. In addition, experimental data are lacking on arrhythmia occurrence after BM cell injection. We hypothesized that therapeutic efficacy and arrhythmia occurrence induced by BM cell injection may be affected by the cell delivery route. METHODS AND RESULTS: Three weeks after left coronary artery ligation, wild-type female rats were injected with 1x10(7) mononuclear BM cells derived from green fluorescent protein-transgenic male rats through either a direct intramyocardial or a retrograde intracoronary route. Both intramyocardial and intracoronary injection of BM cells demonstrated similar improvement in left ventricular ejection fraction measured by echocardiography and a similar graft size analyzed by real-time polymerase chain reaction for the Y chromosome-specific Sry gene. Noticeably, intramyocardial injection of BM cells induced frequent ventricular premature contractions (108+/-73 per hour at 7 days after BM cell injection), including multiform, consecutive ventricular premature contractions and ventricular tachycardia for the initial 14 days; intracoronary injection of BM cells and intramyocardial injection of phosphate-buffered saline rarely induced arrhythmias. Immunohistochemistry demonstrated that intramyocardial BM cell injection formed distinct cell clusters containing donor-derived cells and accumulated host-derived inflammatory cells in the infarct border zone, whereas intracoronary BM cell injection provided more homogeneous donor cell dissemination with less inflammation and without disrupting the native myocardial structure. CONCLUSIONS: BM cell injection is able to improve cardiac function in ischemic chronic heart failure but has a risk of arrhythmia occurrence when the intramyocardial route is used. Such arrhythmias may be prevented by using the intracoronary route. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17438152&query_hl=1 ER - TY - JFULL T1 - Role of DNA methylation in stable gene repression. A1 - Lande-Diner, L A1 - Zhang, J A1 - Ben-Porath, I A1 - Amariglio, N A1 - Keshet, I A1 - Hecht, M A1 - Azuara, V A1 - Fisher, AG A1 - Rechavi, G A1 - Cedar, H J1 - J Biol Chem Y1 - 2007/04/20/ VL - 282 SN - 0021-9258 SP - 12194 EP - 12200 N2 - A large fraction of the animal genome is maintained in a transcriptionally repressed state throughout development. By generating viable Dnmt1(-)(/)(-) mouse cells we have been able to study the effect of DNA methylation on both gene expression and chromatin structure. Our results confirm that the underlying methylation pattern has a profound effect on histone acetylation and is the major effector of me-H3(K4) in the animal genome. We demonstrate that many methylated genes are subject to additional repression mechanisms that also impact on histone acetylation, and the data suggest that late replication timing may play an important role in this process. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17311920&query_hl=1 ER - TY - JFULL T1 - Targeted therapy for inherited GPI deficiency. A1 - Almeida, AM A1 - Murakami, Y A1 - Baker, A A1 - Maeda, Y A1 - Roberts, IA A1 - Kinoshita, T A1 - Layton, DM A1 - Karadimitris, A J1 - N Engl J Med Y1 - 2007/04/19/ VL - 356 SN - 1533-4406 SP - 1641 EP - 1647 N2 - Disrupted binding of the transcription factor Sp1 to the mutated promoter region of the mannosyl transferase-encoding gene PIGM causes inherited glycosylphosphatidylinositol (GPI) deficiency characterized by splanchnic vein thrombosis and epilepsy. We show that this results in histone hypoacetylation at the promoter of PIGM. The histone deacetylase inhibitor butyrate increases PIGM transcription and surface GPI expression in vitro as well as in vivo through enhanced histone acetylation in an Sp1-dependent manner. More important, the drug caused complete cessation of intractable seizures in a child with inherited GPI deficiency. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17442906&query_hl=1 ER - TY - JFULL T1 - Potential of CD34 in the regulation of symmetrical and asymmetrical divisions by hematopoietic progenitor cells. A1 - Bullock, TE A1 - Wen, B A1 - Marley, SB A1 - Gordon, MY J1 - Stem Cells Y1 - 2007/04// VL - 25 SN - 1066-5099 SP - 844 EP - 851 N2 - The control of symmetric and asymmetric division in the hematopoietic stem/progenitor cell population is critically important for the regulation of blood cell production. Asymmetric divisions depend on cell polarization, which may be conferred by location and/or interaction with neighboring cells. In this study, we sought evidence for polarization in CD34+ cells, which interact by binding to one another. In these cells, surface molecules became redistributed by mechanisms that included transport by lipid rafts, and the interacting cells were able to communicate via gap junctions. These changes were accompanied by modulation of cell cycle regulating proteins (p16(Ink4a), p27(kip1), cyclins D, and the retinoblastoma pathway proteins) and a reduction in progenitor cell proliferation in vitro. These results are consistent with an increase in asymmetric cell division kinetics. Accordingly, we found that interaction between CD34+ cells influenced the plane of cell division in a way that suggests unequal sharing of Notch-1 between daughter cell progeny. We conclude that interaction between CD34+ cells may coordinate cell function and participate in the control of hematopoietic stem/progenitor cell division kinetics. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17185613&query_hl=1 ER - TY - JFULL T1 - Enhanced differentiation and mineralization of human fetal osteoblasts on PDLLA containing Bioglass composite films in the absence of osteogenic supplements. A1 - Tsigkou, O A1 - Hench, LL A1 - Boccaccini, AR A1 - Polak, JM A1 - Stevens, MM J1 - J Biomed Mater Res A Y1 - 2007/03/15/ VL - 80 SN - 1549-3296 SP - 837 EP - 851 N2 - This study investigates the cellular response of fetal osteoblasts to bioactive resorbable composite films consisting of a poly-D,L-lactide (PDLLA) matrix and bioactive glass 45S5 Bioglass (BG) particles at three different concentrations (0% (PDLLA), 5% (P/BG5), and 40% (P/BG40)). Using scanning electron microscopy (SEM) we observed that cells were less spread and elongated on PDLLA and P/BG5, whereas cells on P/BG40 were elongated but with multiple protrusions spreading over the BG particles. Vinculin immunostaining revealed similar distribution of focal adhesion contacts on all cells independent of substratum, indicating that all materials permitted cell adhesion. However, when differentiation and maturation of fetal osteoblasts was examined, incorporation of 45S5 BG within the PDLLA matrix was found to significantly (p < 0.05) enhance alkaline phosphatase enzymatic activity and osteocalcin protein synthesis compared to tissue culture polystyrene controls and PDLLA alone. Alizarin red staining indicated extracellular matrix mineralization on both P/BG5 and P/BG40, with significantly more bone nodules formed than on PDLLA. Real time RT-PCR revealed that expression of bone sialoprotein was also affected by the BG containing films compared to controls, whereas expression of Collagen Type I was not influenced. By performing these investigations in the absence of osteogenic factors it appears that the incorporation of BG stimulates osteoblast differentiation and mineralization of the extracellular matrix, demonstrating the osteoinductive capacity of the composite. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17072851&query_hl=1 ER - TY - JFULL T1 - Re: Impact of classification of hilar cholangiocarcinomas (Klatskin tumors) on incidence of intra- and extrahepatic cholangiocarcinorna in the United States A1 - Matull, WR A1 - Khan, SA A1 - Pereira, SP J1 - J NATL CANCER I Y1 - 2007/03/07/ VL - 99 SN - 0027-8874 SP - 407 EP - 407 ER - TY - JFULL T1 - Extracellular matrix formation and mineralization on a phosphate-free porous bioactive glass scaffold using primary human osteoblast (HOB) cells. A1 - Jones, JR A1 - Tsigkou, O A1 - Coates, EE A1 - Stevens, MM A1 - Polak, JM A1 - Hench, LL J1 - Biomaterials Y1 - 2007/03// VL - 28 SN - 0142-9612 SP - 1653 EP - 1663 N2 - Sol-gel derived bioactive glasses of the 70S30C (70mol% SiO2, 30mol% CaO) composition have been foamed to produce 3D bioactive scaffolds with hierarchical interconnected pore morphologies similar to trabecular bone. The aim of this study was to investigate primary human osteoblast response to porous bioactive glass scaffolds. The scaffolds supported osteoblast growth and induced differentiation, within the 3-week culture period, as depicted by enhanced ALPase enzymatic activity, without the addition of supplementary factors such as ascorbic acid, beta-glycerophosphate and dexamethasone. This is the first time this has been observed on a bioactive glass that does not contain phosphate. Deposition of extracellular matrix was also confirmed by enhanced production of the extracellular matrix protein collagen type I. SEM showed indications of mineralized bone nodule formation without the addition of growth factors. The 70S30C bioactive glass scaffolds therefore fulfil many of the criteria for an ideal scaffold for bone tissue engineering applications. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17175022&query_hl=1 ER - TY - JFULL T1 - The role of the cardiac Na+/Ca2+ exchanger in reverse remodeling: relevance for LVAD-recovery. A1 - Terracciano, CM A1 - Koban, MU A1 - Soppa, GK A1 - Siedlecka, U A1 - Lee, J A1 - Stagg, MA A1 - Yacoub, MH J1 - Ann N Y Acad Sci Y1 - 2007/03// VL - 1099 SN - 0077-8923 SP - 349 EP - 360 N2 - Different strategies can, at least in certain conditions, prevent or reverse myocardial remodeling due to heart failure and induce myocardial functional improvement. Na+/Ca2+ exchanger (NCX) is considered a major player in the pathophysiology of heart failure but its role in reverse remodeling is unknown. A combination of mechanical unloading by left ventricular assist devices (LVADs) and pharmacological therapy has been shown to induce clinical recovery in a limited number of patients with end-stage heart failure. In myocytes isolated from these patients we found that, after LVAD treatment, NCX1/SERCA2a mRNA was 38% higher than at device implant. We studied the ability of NCX to extrude Ca2+ during caffeine-induced SR Ca2+ release in isolated ventricular myocytes from these patients. The time constant of decline was slower in heart failure. In myocytes from patients with clinical recovery following mechanical and pharmacological treatment, NCX1-mediated Ca2+ extrusion was faster compared with myocytes from patient who, despite identical treatment, did not recover. We propose that increased NCX function may be associated with reverse remodeling in patients and that factors that regulate NCX function (i.e., phosphorylation or intracellular [Na+]) other than NCX expression levels alone, may have detrimental consequences on cardiac function. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17446475&query_hl=1 ER - TY - JFULL T1 - Ribosome-lamella complex precursors in acute monocytic leukemia: a study of 6 cases. A1 - Ru, YX A1 - Mi, YC A1 - Liu, JH A1 - Cui, W A1 - Wang, HJ A1 - Zhao, SX A1 - Jian-Xiang, W J1 - Ultrastruct Pathol Y1 - 2007/03// VL - 31 SN - 1521-0758 SP - 135 EP - 140 N2 - The ribosome-lamella complex (RLC) is a cylindrical structure composed of annular lamella associated particles, regarded as ribosomes, around a central core, which is best known in hairy cell leukemia. RLC has been presumed to originate from aggregating rER and ribosomes. Incomplete and maturing RLC structures have been called RLC precursors (pre-RLC). The present paper investigates the various architectural aspects of pre-RLC and the ultrastructural characteristics of the blasts in 6 cases of acute monocytic leukemia (M5) in which these structures occur. Blasts bearing pre-RLC contained irregular nuclei with less heterochromatin and a prominent nucleolus, and many cytoplasmic organelles in an abundant cytoplasm. The findings indicate that pre-RLC might result from an asymmetrical differentiation of organelles in blasts associated with expression of CD117 and CD56 but default of CD14 in M5. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17613993&query_hl=1 ER - TY - JFULL T1 - Conventional Western blotting techniques will not reliably quantify p210(BCR-ABL1) levels in CML mononuclear cells A1 - Patel, H A1 - Marley, SB A1 - Gordon, MY J1 - BLOOD Y1 - 2007/02/01/ VL - 109 SN - 0006-4971 SP - 1335 EP - 1335 ER - TY - JFULL T1 - The human embryonic stem cell-derived cardiomyocyte as a pharmacological model. A1 - Harding, SE A1 - Ali, NN A1 - Brito-Martins, M A1 - Gorelik, J J1 - Pharmacol Ther Y1 - 2007/02// VL - 113 SN - 0163-7258 SP - 341 EP - 353 N2 - Embryonic stem (ES) cells are specialised cells derived from the early embryo, which are capable of both sustained propagation in the undifferentiated state as well as subsequent differentiation into the majority of cell lineages. Human ES cells are being developed for clinical tissue repair, but a number of problems must be addressed before this becomes a reality. However, they also have potential for translational benefit through its use as a test system for screening pharmaceutical compounds. In the cardiac field, present model systems are not ideal for either screening or basic pharmacological/physiological studies. Cardiomyocytes produced from human ES differentiation have advantages for these purposes over the primary isolated cells or the small number of cell lines available. This review describes the methodology for obtaining cardiomyocytes from human embryonic stem cell-derived cardiomyocyte (hESCM), for increasing the proportion of cardiomyocytes in the preparation and for isolating single embryonic stem cell-derived cardiomyocyte (ESCM) from clusters. Their morphological, contractile and electrophysiological characteristics are compared to mature and immature primary cardiomyocytes. The advantages and disadvantages of the hESCM preparation for long term culture and genetic manipulation are described. Basic pharmacological studies on adrenoceptors and muscarinic receptors in hESCM have been performed, and have given stable and reproducible responses. Prolongation of repolarisation can be detected using hESCM cultured on multielectrode arrays (MEA). Human ESCM have a clear potential to improve model systems available for both basic scientific studies and pharmaceutical screening of cardiac target compounds. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17081613&query_hl=1 ER - TY - JFULL T1 - Stem cells as a treatment for chronic liver disease and diabetes. A1 - Levicar, N A1 - Dimarakis, I A1 - Flores, C A1 - Tracey, J A1 - Gordon, MY A1 - Habib, NA J1 - Handb Exp Pharmacol Y1 - 2007/// SN - 0171-2004 SP - 243 EP - 262 N2 - Advances in stem cell biology and the discovery of pluripotent stem cells have made the prospect of cell therapy and tissue regeneration a clinical reality. Cell therapies hold great promise to repair, restore, replace or regenerate affected organs and may perform better than any pharmacological or mechanical device. There is an accumulating body of evidence supporting the contribution of adult stem cells, in particular those of bone marrow origin, to liver and pancreatic islet cell regeneration. In this review, we will focus on the cell therapy for the diseased liver and pancreas by adult haematopoietic stem cells, as well as their possible contribution and application to tissue regeneration. Furthermore, recent progress in the generation, culture and targeted differentiation of human haematopoietic stem cells to hepatic and pancreatic lineages will be discussed. We will also explore the possibility that stem cell technology may lead to the development of clinical modalities for human liver disease and diabetes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17554512&query_hl=1 ER - TY - JFULL T1 - Cost-utility analysis in china: differences and difficulties compared with developed countries. A1 - Bao Peng, L A1 - Qing Tan, C A1 - Min Wan, X A1 - Cui, W J1 - Pharmacoeconomics Y1 - 2007/// VL - 25 SN - 1170-7690 SP - 619 EP - 619 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17610341&query_hl=1 ER - TY - JFULL T1 - Direct differentiation of human embryonic stem cells to hepatocyte-like cells exhibiting functional activities. A1 - Hay, DC A1 - Zhao, D A1 - Ross, A A1 - Mandalam, R A1 - Lebkowski, J A1 - Cui, W J1 - Cloning Stem Cells Y1 - 2007/// VL - 9 SN - 1536-2302 SP - 51 EP - 62 N2 - The utilization of human hepatocytes for biomedical research, drug discovery, and treatment of liver diseases is hindered by the limited availability of donated livers and the variability of their derived hepatocytes. Human embryonic stem cells (hESCs) are pluripotent and provide a unique, unlimited resource for human hepatocytes. However, differentiation of hESCs to hepatocytes remains a challenge. We have developed a multistage procedure by which hESCs can be directly differentiated to hepatocyte-like cells without embryoid body formation and the requirement of sodium butyrate. The hESC-derived hepatocyte-like cells (HLCs) exhibited characteristic hepatocyte morphology, expressed hepatocyte markers, including alpha-fetoprotein, albumin, and hepatocyte nuclear factor 4alpha, and possessed hepatocyte-specific activities, such as p450 metabolism, albumin production, glycogen storage, and uptake and excretion of indocyanine green. Hepatocyte growth factor was found to play a positive role in promoting hepatocyte differentiation. Our differentiation system has shown that hESCs can be differentiated to hepatocyte-like cells capable of executing a range of hepatocyte functions. Therefore, it presents a proof-of-principle of potential applications of using the hESC-derived hepatocytes. Additionally, the hESC-derived HLCs provide a unique model to study the mechanisms involved in human hepatocyte differentiation and liver function. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17386014&query_hl=1 ER - TY - JFULL T1 - A point mutation in an Sp1 binding motif in the promoter of the mannosyltransferase-encoding PIG-M gene causes inherited glycosylphosphatidylinositol deficiency A1 - Murakarni, Y A1 - Almeida, A A1 - Layton, M A1 - Hillmen, P A1 - Maeda, Y A1 - Karadimitris, A A1 - Kinoshita, T J1 - MOL IMMUNOL Y1 - 2007/01// VL - 44 SN - 0161-5890 SP - 218 EP - 218 ER - TY - JFULL T1 - The nuclear receptor co-repressor RIP140 controls the expression of metabolic gene networks. A1 - Parker, MG A1 - Christian, M A1 - White, R J1 - Biochem Soc Trans Y1 - 2006/12// VL - 34 SN - 0300-5127 SP - 1103 EP - 1106 N2 - NRs (nuclear receptors) regulate the expression of specific gene networks in target cells by recruiting cofactor complexes involved in chromatin remodelling and in the assembly of transcription complexes. The importance of activating gene expression, in metabolic tissues, is well established, but the contribution of transcriptional inhibition is less well defined. In this review, we highlight a crucial role for RIP140 (receptor-interacting protein 140), a transcriptional co-repressor for NR, in the regulation of metabolic gene expression. Many genes involved in lipid and carbohydrate metabolism are repressed by RIP140 in adipose and muscle. The repressive function of RIP140 results from its ability to bridge NRs to repressive enzyme complexes that modify DNA and histones. In the absence of RIP140, expression from many metabolic genes is increased so that mice exhibit a lean phenotype and resistance to high-fat-diet-induced obesity and display increased glucose tolerance and insulin sensitivity. We propose that a functional interplay between transcriptional activators and the co-repressor RIP140 is an essential process in metabolic regulation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17073760&query_hl=1 ER - TY - JFULL T1 - A novel Z-groove index characterizing myocardial surface structure. A1 - Gorelik, J A1 - Yang, LQ A1 - Zhang, Y A1 - Lab, M A1 - Korchev, Y A1 - Harding, SE J1 - Cardiovasc Res Y1 - 2006/12/01/ VL - 72 SN - 0008-6363 SP - 422 EP - 429 N2 - OBJECTIVE: The role of t-tubule structures in excitation-contraction coupling of ventricular myocytes has been investigated by disruption using prolonged culture, or osmotic shock with formamide. We have used a new method, the Scanning Ion Conductance Microscope (SICM), to investigate in more detail the changes in surface structure of live myocytes during these interventions and to relate them to contractile effects. METHODS: Freshly isolated ventricular myocytes from adult rat hearts were either incubated with formamide, then washed to produce osmotic shock, or put into culture for 2, 4 and 7 days. Contractile characteristics of single myocytes were then measured using the IonOptix system, and in parallel imaged using the SICM which produces a 3-dimensional topographical representation of the cell surface. Loss of t-tubules was quantitated with confocal microscopy after staining with the membrane dye di-8-ANNEPS, and sarcomere structure revealed by immunocytochemical detection of alpha-actinin. RESULTS: Detubulation was produced by either method, with formamide equivalent to 4 day culture in quantitative measures of ANNEPS t-tubule/membrane ratio. SICM images confirmed the loss of t-tubule indentations. Disruption of the Z-groove structure and flattening of the surface were also noted with formamide and, to a lesser extent, culture. A novel Z-groove index was introduced to describe this effect more quantitatively. Contraction and relaxation were impaired by the detubulation methods, but formamide had a markedly greater depressant effect on contraction amplitude than equivalent detubulation by culture. CONCLUSION: Changes in contraction amplitude after detubulation with formamide were more closely related to the alteration in Z-groove structure than loss of t-tubules alone. As well as disrupting t-tubule-induced excitation and calcium movements, formamide may alter the transmission of contraction in the myocyte by interference with sarcomere attachment at the Z-line. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17054929&query_hl=1 ER - TY - JFULL T1 - International Union of Pharmacology. LXIV. Estrogen receptors A1 - Dahlman-Wright, K A1 - Cavailles, V A1 - Fuqua, SA A1 - Jordan, VC A1 - Katzenellenbogen, JA A1 - Korach, KS A1 - Maggi, A A1 - Muramatsu, M A1 - Parker, MG A1 - Gustafsson, JA J1 - PHARMACOL REV Y1 - 2006/12// VL - 58 SN - 0031-6997 SP - 773 EP - 781 ER - TY - JFULL T1 - Detection in primary chronic myeloid leukaemia cells of p210BCR-ABL1 in complexes with adaptor proteins CBL, CRKL, and GRB2. A1 - Patel, H A1 - Marley, SB A1 - Gordon, MY J1 - Genes Chromosomes Cancer Y1 - 2006/12// VL - 45 SN - 1045-2257 SP - 1121 EP - 1129 N2 - Chronic myeloid leukemia (CML) arises as a consequence of the expression of a chimeric fusion protein, p210BCR-ABL1, which is localized to the cytoplasm and has constitutive protein tyrosine kinase activity. Extensive publications report that p210BCR-ABL1 complexed with multiple cytoplasmic proteins can modulate several cell signaling pathways. However, while altered signaling states can be demonstrated in primary CML material, most of the reported analytical work on complexed proteins has been done in cell lines expressing p210BCR-ABL1. This has been necessary because primary hemopoietic cell lysates contain a degradative activity which rapidly and permanently destroys p210BCR-ABL1, precluding accurate p210BCR-ABL1 quantification by Western blotting or investigation of coimmunoprecipitating proteins in primary cells. This degradative activity has proven intractable to inhibition by conventional protease inhibitors. We show here that the degradative activity in primary cells is associated with cell lysosomes and is most likely to be an acid-dependent hydrolase. By lysing primary hemopoietic cells at high pH, we have demonstrated substantial inhibition of the p210BCR-ABL1-degradative activity and now report, to the best of our knowledge, the first published demonstration by coimmunoprecipitation of the association between p210BCR-ABL1 and cytoplasmic effector proteins in primary CML material. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16955467&query_hl=1 ER - TY - JFULL T1 - Telomerase with mutated catalytic motifs has dominant negative effects on telomerase activity and inhibits cell growth. A1 - Rahman, R A1 - Mo, L A1 - Cui, W J1 - Biochem Biophys Res Commun Y1 - 2006/11/24/ VL - 350 SN - 0006-291X SP - 796 EP - 802 N2 - Telomerase catalytic subunit (TERT) seems a key factor controlling telomerase activity, telomere length, and cell growth. To further address this issue, we forced expression of a catalytically inactive mutant human TERT (hTERT) in hTERT-immortalised sheep fibroblasts to examine its effects. Expression of mutant hTERT compromised telomerase activity reconstituted by wild-type hTERT in a manner directly attributable to mutant hTERT expression level. High levels of mutant hTERT expression inhibited cell growth with a subset of cells entering replicative senescence. Furthermore, significant telomere attrition was evident in two of three clones with high levels of mutant hTERT expression. Our findings are consistent with the notion that hTERT homodimers are necessarily required to form a functional telomerase complex at the telomere substrate. We also highlight the requirement of a more thorough understanding of telomerase- and telomere-associated factors to understand fully the interplay that governs telomere homeostasis in vitro and in vivo. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17026956&query_hl=1 ER - TY - JFULL T1 - Constitutive expression of SALL4B in mice confers properties of leukemic stem cells to committed hematopoietic progenitors. A1 - Chai, L A1 - Cui, W A1 - Ma, Y A1 - Yang, JC A1 - Kong, N A1 - Kao, G A1 - Silberstein, L J1 - BLOOD Y1 - 2006/11/16/ VL - 108 SN - 0006-4971 SP - 75A EP - 75A ER - TY - JFULL T1 - Targeted molecular therapy for inherited glycosylphosphatidylinositol deficiency. A1 - Almeida, AM A1 - Murakami, Y A1 - Baker, A A1 - Maeda, Y A1 - Roberts, I A1 - Kinoshita, T A1 - Layton, DM A1 - Karadimitris, A J1 - BLOOD Y1 - 2006/11/16/ VL - 108 SN - 0006-4971 SP - 148A EP - 148A ER - TY - JFULL T1 - A novel HSV-1 virus, JS1/34.5-/47-, purges contaminating breast cancer cells from bone marrow. A1 - Hu, JC A1 - Booth, MJ A1 - Tripuraneni, G A1 - Davies, D A1 - Zaidi, SA A1 - Tamburo de Bella, M A1 - Slade, MJ A1 - Marley, SB A1 - Gordon, MY A1 - Coffin, RS A1 - Coombes, RC A1 - Kamalati, T J1 - Clin Cancer Res Y1 - 2006/11/15/ VL - 12 SN - 1078-0432 SP - 6853 EP - 6862 N2 - PURPOSE: Oncolytic herpes simplex virus type 1 (HSV-1) vectors show considerable promise as agents for cancer therapy. We have developed a novel recombinant HSV-1 virus (JS1/34.5-/47-) for purging of occult breast cancer cells from bone marrow of patients. Here, we evaluate the therapeutic efficacy of this oncolytic virus. EXPERIMENTAL DESIGN: Electron microscopy was used to determine whether human breast cancer and bone marrow cells are permissive for JS1/34.5-/47- infection. Subsequently, the biological effects of JS1/34.5-/47- infection on human breast cancer cells and bone marrow were established using cell proliferation and colony formation assays, and the efficiency of cell kill was evaluated. Finally, the efficiency of JS1/34.5-/47- purging of breast cancer cells was examined in cocultures of breast cancer cells with bone marrow as well as bone marrow samples from high-risk breast cancer patients. RESULTS: We show effective killing of human breast cancer cell lines with the JS1/34.5-/47- virus. Furthermore, we show that treatment with JS1/34.5-/47- can significantly inhibit the growth of breast cancer cell lines without affecting cocultured mononuclear hematopoietic cells. Finally, we have found that the virus is effective in destroying disseminated tumors cells in bone marrow taken from breast cancer patients, without affecting the hematopoietic contents in these samples. CONCLUSION: Collectively, our data show that the JS1/34.5-/47- virus can selectively target breast cancer cells while sparing hematopoietic cells, suggesting that JS1/34.5-/47- can be used to purge contaminating breast cancer cells from human bone marrow in the setting of autologous hematopoietic cell transplantation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17121907&query_hl=1 ER - TY - JFULL T1 - Suppression of receptor interacting protein 140 repressive activity by protein arginine methylation A1 - Huq, MDM A1 - Gupta, P A1 - Tsai, NP A1 - White, R A1 - Parker, MG A1 - Wei, LN J1 - EMBO J Y1 - 2006/11/01/ VL - 25 SN - 0261-4189 SP - 5094 EP - 5104 N2 - Receptor interacting protein 140 (RIP140), a ligand-dependent corepressor for nuclear receptors, can be modified by arginine methylation. Three methylated arginine residues, at Arg-240, Arg-650, and Arg-948, were identified by mass spectrometric analysis. Site-directed mutagenesis studies demonstrated the functionality of these arginine residues. The biological activity of RIP140 was suppressed by protein arginine methyltransferase 1 (PRMT1) due to RIP140 methylation, which reduced the recruitment of histone deacetylases to RIP140 and facilitated its nuclear export by enhancing interaction with exportin 1. A constitutive negative (Arg/Ala) mutant of RIP140 was resistant to the effect of PRMT1, and a constitutive positive (Arg/Phe) mutation mimicked the effect of arginine methylation. The biological activities of the wild type and the mutant proteins were examined in RIP140-null MEF cells. This study uncovered a novel means to inactivate, or suppress, RIP140, and demonstrated protein arginine methylation as a critical type of modification for corepressor. ER - TY - JFULL T1 - Historic and current strategies in bone tissue engineering: do we have a hope in Hench? A1 - Gentleman, E A1 - Polak, JM J1 - J Mater Sci Mater Med Y1 - 2006/11// VL - 17 SN - 0957-4530 SP - 1029 EP - 1035 N2 - Professors Larry Hench and Julia Polak formed the Tissue Engineering and Regenerative Medicine Centre (TERM) at Imperial College London to foster collaborations between biologists and materials scientists. Early work at the center elucidated the biomolecular interactions between primary human osteoblasts and 45S5 Bioglass . As research efforts expanded, the team discovered that the dissolution products of both 45S5 Bioglass and 58S sol-gel bioactive glasses had osteoblastic stimulatory properties. To address the shortage of appropriate cells for bone tissue engineering applications, TERM scientists also demonstrated the differentiation of embryonic stem (ES) cells to osteoblasts when treated with the dissolution products of bioactive glasses. They also found that the soluble factors ascorbic acid, beta -glycerophosphate, and dexamethasone preferentially differentiated ES cells to osteoblasts, and their combination with the dissolution products of bioactive glasses stimulated differentiation even further. Taken together, these results demonstrate the suitability of bioactive glasses as scaffolds for bone tissue engineering as they not only provide an osteoconductive and osteoproductive substrate, but also actively stimulate cells to express appropriate osteoblastic phenotypes. Professor Hench's vision to pioneer regenerative medicine research continues with the aim of developing novel therapeutics to treat musculoskeletal disability. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17122915&query_hl=1 ER - TY - JFULL T1 - The effect of Ischemia/Reperfusion on the pacing threshold by transcoronary pacing A1 - Cui, W A1 - Liu, F A1 - Xie, RQ A1 - Lu, J A1 - Li, BH A1 - Wu, JF A1 - Yang, XC A1 - Hao, YM A1 - Du, J J1 - CIRCULATION Y1 - 2006/10/31/ VL - 114 SN - 0009-7322 SP - 691 EP - 691 ER - TY - JFULL T1 - Apelin, the ligand for the angiotensin receptor-like 1, directly affects cardiomyocyte contractility and electrophysiology A1 - Farkasfalvi, K A1 - Stagg, MA A1 - Siedlecka, U A1 - Lee, J A1 - Soppa, GK A1 - Marczin, N A1 - Szokodi, I A1 - Yacoub, MH A1 - Terracciano, CM J1 - CIRCULATION Y1 - 2006/10/31/ VL - 114 SN - 0009-7322 SP - 300 EP - 300 ER - TY - JFULL T1 - The apelin receptor mRNA levels and myocardial recovery in end-stage heart failure patients treated with left ventricular assist devices (LVADs) A1 - Farkasfalvi, K A1 - Felkin, L A1 - Latif, N A1 - Soppa, GK A1 - George, R A1 - Birks, E A1 - Barton, P A1 - Marczin, N A1 - Yacoub, MH A1 - Terracciano, CM J1 - CIRCULATION Y1 - 2006/10/31/ VL - 114 SN - 0009-7322 SP - 484 EP - 484 ER - TY - JFULL T1 - Chronic clenbuterol treatment improves heart and myocyte function in heart failure, with and without mechanical unloading. A1 - Soppa, GK A1 - Lee, J A1 - Stagg, MA A1 - Youssef, S A1 - Siedlecka, U A1 - Yacoub, MH A1 - Terracciano, CM J1 - CIRCULATION Y1 - 2006/10/31/ VL - 114 SN - 0009-7322 SP - 483 EP - 483 ER - TY - JFULL T1 - Skeletal myoblasts and bone marrow-derived cells enhance the contractile function of isolated failing cardiomyocytes in co-culture A1 - Lee, J A1 - Stagg, MA A1 - Soppa, GK A1 - Siedlecka, U A1 - Yacoub, MH A1 - Terracciano, CM J1 - CIRCULATION Y1 - 2006/10/31/ VL - 114 SN - 0009-7322 SP - 93 EP - 93 ER - TY - JFULL T1 - RIP140 expression is stimulated by estrogen-related receptor alpha during adipogenesis. A1 - Nichol, D A1 - Christian, M A1 - Steel, JH A1 - White, R A1 - Parker, MG J1 - J Biol Chem Y1 - 2006/10/27/ VL - 281 SN - 0021-9258 SP - 32140 EP - 32147 N2 - RIP140 is a corepressor for nuclear receptors that regulates energy expenditure in adipose tissue by suppressing the expression of clusters of metabolic genes involved in glucose and lipid metabolism. The gene encoding RIP140/Nrip1 contains only one coding exon but has multiple promoters and 5' non-coding exons that are subject to alternative splicing. In adipocytes we have defined a promoter, referred to as P2, that is preferentially utilized and activated during adipogenesis. Expression studies and chromatin immunoprecipitation experiments indicate that estrogen-related receptor alpha (ERRalpha), the level of which increases during adipogenesis in parallel with RIP140, stimulates transcription from the P2 promoter. Further analysis indicates that ERRalpha is capable of activating RIP140 gene transcription by two mechanisms, directly by binding to an estrogen receptor element/ERR element at -650/-633 and indirectly through Sp1 binding sites in the proximal promoter. Thus, the up-regulation of RIP140 by ERRalpha during adipogenesis may provide an inhibitory feedback mechanism to control the expression of many nuclear receptor target genes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16923809&query_hl=1 ER - TY - JFULL T1 - The effects of unfractionated heparin and low-molecular-weight heparin on platelet activation A1 - Cui, W A1 - Lu, JC A1 - Guo, N A1 - Liu, F A1 - Xie, RQ A1 - Gu, GQ A1 - Du, J J1 - AM J CARDIOL Y1 - 2006/10/22/ VL - 98 SN - 0002-9149 SP - 195M EP - 195M ER - TY - JFULL T1 - Biocompatibility of poly-DL-lactic acid (PDLLA) for lung tissue engineering. A1 - Lin, YM A1 - Boccaccini, AR A1 - Polak, JM A1 - Bishop, AE A1 - Maquet, V J1 - J Biomater Appl Y1 - 2006/10// VL - 21 SN - 0885-3282 SP - 109 EP - 118 N2 - This study explores the possibility of growing lung cells on poly-DL-lactic acid (PDLLA) scaffolds, with a view to in future engineer pulmonary tissue for human implantation. As a first step in this process, the ability of PDLLA to maintain the growth of lung epithelium is tested using a robust cell line. Poly-DL-lactic acid has been investigated in two forms, as planar discs and as 3-D foams, and it has been demonstrated that PDLLA is not only nontoxic to pneumocytes but it also actively supports their growth. The initial findings suggest that the material is an appropriate matrix for engineering of distal lung tissue. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16443629&query_hl=1 ER - TY - JFULL T1 - Effects and mechanisms of pentoxitylline on a HEPG2 steatosis model A1 - Chen, S A1 - Cui, W A1 - Hu, KQ J1 - HEPATOLOGY Y1 - 2006/10// VL - 44 SN - 0270-9139 SP - 667A EP - 668A ER - TY - JFULL T1 - In vivo effects and mechanisms of silibinin on human hepatoma (HuH7) xenografts in nude mice A1 - Cui, W A1 - Hu, KQ J1 - HEPATOLOGY Y1 - 2006/10// VL - 44 SN - 0270-9139 SP - 526A EP - 527A ER - TY - JFULL T1 - The SWI/SNF chromatin remodeling subunit BAF57 is a critical regulator of estrogen receptor function in breast cancer cells. A1 - García-Pedrero, JM A1 - Kiskinis, E A1 - Parker, MG A1 - Belandia, B J1 - J Biol Chem Y1 - 2006/08/11/ VL - 281 SN - 0021-9258 SP - 22656 EP - 22664 N2 - Estrogen receptors (ERs) play critical roles in both normal mammary gland development and in the formation and progression of breast tumors, constituting a major therapeutic target for breast cancer treatment. We have previously described that ER transcriptional activity is potentiated by BAF57, a core subunit of the mammalian SWI/SNF chromatin remodeling complex. Here we provide evidence demonstrating an important role for BAF57 as regulator of ER functions in breast cancer cells. Different experimental manipulations leading to the abrogation of BAF57 expression and/or function severely reduced the expression of various endogenous ER target genes and blocked estrogen-stimulated proliferation in ZR-75-1 breast cancer cells. Moreover, using a structure-function analysis, we have defined the protein domains required for the functional interaction between ERalpha and BAF57, including a key region within the hinge of ERalpha that is essential for BAF57 recruitment and its function on ER-mediated transcription. Interestingly, we found that BAF57 is an ER subtype-selective modulator that specifically regulates ERalpha-mediated transcription. Taken together, our results suggest that targeting BAF57 could represent a new way to effectively inhibit the action of ERalpha. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16769725&query_hl=1 ER - TY - JFULL T1 - Anaemia of unknown origin in chronic heart failure is a consequence of immune-mediated suppression of erythroid colony formation and erythropoietin resistance A1 - Okonko, DO A1 - Marley, SB A1 - Crosato, M A1 - Anker, SD A1 - Gordon, MY A1 - Poole-Wilson, PA J1 - EUR HEART J Y1 - 2006/08// VL - 27 SN - 0195-668X SP - 482 EP - 483 ER - TY - JFULL T1 - Natural killer T cells and haemopoiesis. A1 - Karadimitris, A A1 - Patterson, S A1 - Spanoudakis, E J1 - Br J Haematol Y1 - 2006/08// VL - 134 SN - 0007-1048 SP - 263 EP - 272 N2 - Invariant natural killer T (iNKT) cells are a small but powerful subset of regulatory T cells involved in the modulation of a variety of normal and pathological immune responses. In contrast to conventional or other types of regulatory T cells, they are activated by glycolipid and phospholipid ligands that are presented to them by the non-polymorphic, major histocompatibility complex class I-like molecule CD1d. The in-depth understanding of their function has resulted in successful, iNKT cell-centred experimental therapeutic interventions including prevention of graft-versus-host disease and anti-leukaemia effects. Extending these successes into the clinical arena will require better understanding of their contribution to the pathogenesis of human, including haematological, diseases. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16848769&query_hl=1 ER - TY - JFULL T1 - Stem cells--potential for repairing damaged lungs and growing human lungs for transplant. A1 - Bishop, AE A1 - Rippon, HJ J1 - Expert Opin Biol Ther Y1 - 2006/08// VL - 6 SN - 1744-7682 SP - 751 EP - 758 N2 - Repair or regeneration of defective lung epithelium would be of great therapeutic potential. It is estimated by the British Lung Foundation that 1 in 7 people in the UK is affected by a lung disease and that 1 in 4 admissions to children's wards are as a result of respiratory problems. Potential cellular sources for the regeneration of lung tissue in vivo or lung tissue engineering in vitro include endogenous pulmonary epithelial stem cells, extrapulmonary circulating stem cells and embryonic stem cells. This article discusses the potential role of each of these stem cell types in future approaches to the treatment of lung injury and disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16856797&query_hl=1 ER - TY - JFULL T1 - Metabolic regulation by the nuclear receptor corepressor RIP140. A1 - Christian, M A1 - White, R A1 - Parker, MG J1 - Trends Endocrinol Metab Y1 - 2006/08// VL - 17 SN - 1043-2760 SP - 243 EP - 250 N2 - Whereas the importance of activating gene expression in metabolic pathways to control energy homeostasis is well established, the contribution of transcriptional inhibition is less well defined. In this review we highlight a crucial role of RIP140, a transcriptional corepressor for nuclear receptors, in the regulation of energy expenditure. Mice devoid of the RIP140 gene are lean, exhibit resistance to high-fat-diet-induced obesity, and have increased glucose tolerance and insulin sensitivity. Consistent with these observations, RIP140 suppresses the expression of gene clusters that are involved in lipid and carbohydrate metabolism, including fatty acid oxidation, oxidative phosphorylation and mitochondrial uncoupling. Therefore, the functional interplay between transcriptional activators and the corepressor RIP140 is an essential process in metabolic regulation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16815031&query_hl=1 ER - TY - JFULL T1 - Iron particles for noninvasive monitoring of bone marrow stromal cell engraftment into, and isolation of viable engrafted donor cells from, the heart A1 - Stuckey, DJ A1 - Carr, CA A1 - Martin-Rendon, E A1 - Tylera, DJ A1 - Willmott, C A1 - Cassidy, PJ A1 - Hale, SJM A1 - Schneider, JE A1 - Tatton, L A1 - Harding, SE A1 - Radda, GK A1 - Watt, S A1 - Clarke, K J1 - STEM CELLS Y1 - 2006/08// VL - 24 SN - 1066-5099 SP - 1968 EP - 1975 N2 - Stem cells offer a promising approach to the treatment of myocardial infarction and prevention of heart failure. We have used iron labeling of bone marrow stromal cells (BMSCs) to noninvasively track cell location in the infarcted rat heart over 16 weeks using cine-magnetic resonance imaging (cine-MRI) and to isolate the BMSCs from the grafted hearts using the magnetic properties of the donor cells. BMSCs were isolated from rat bone marrow, characterized by flow cytometry, transduced with lentiviral vectors expressing green fluorescent protein (GFP), and labeled with iron particles. BMSCs were injected into the infarct periphery immediately following coronary artery ligation, and rat hearts were imaged at 1, 4, 10, and 16 weeks postinfarction. Signal voids caused by the iron particles in the BMSCs were detected in all rats at all time points. In mildly infarcted hearts, the volume of the signal void decreased over the 16 weeks, whereas the signal void volume did not decrease significantly in severely infarcted hearts. High-resolution three-dimensional magnetic resonance (MR) microscopy identified hypointense regions at the same position as in vivo. Donor cells containing iron particles and expressing GFP were identified in MR-targeted heart sections after magnetic cell separation from digested hearts. In conclusion, MRI can be used to track cells labeled with iron particles in damaged tissue for at least 16 weeks after injection and to guide tissue sectioning by accurately identifying regions of cell engraftment. The magnetic properties of the iron-labeled donor cells can be used for their isolation from host tissue to enable further characterization. ER - TY - JFULL T1 - Structural analogues of AMD3100 mobilise haematopoietic progenitor cells from bone marrow in vivo according to their ability to inhibit CXCL12 binding to CXCR4 in vitro. A1 - Martin, C A1 - Bridger, GJ A1 - Rankin, SM J1 - Br J Haematol Y1 - 2006/08// VL - 134 SN - 0007-1048 SP - 326 EP - 329 N2 - The CXCR4 antagonist, AMD3100, stimulates a rapid increase in circulating numbers of haematopoeitic progenitor cells (HPCs) in both mice and human healthy volunteers. An in situ perfusion system of the mouse femoral bone marrow was used to provide the first direct evidence that AMD3100 mobilises HPCs from the bone marrow. Structural analogues of AMD3100 demonstrated that the ability of these compounds to mobilise HPCs in vivo correlated with their capacity to antagonise CXCR4 in vitro. This model system was also used to demonstrate additive effects of AMD3100 administered acutely, with granulocyte colony-stimulating factor administered chronically, with respect to HPC mobilisation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16787495&query_hl=1 ER - TY - JFULL T1 - Antibiotics reduce the growth rate and differentiation of embryonic stem cell cultures. A1 - Cohen, S A1 - Samadikuchaksaraei, A A1 - Polak, JM A1 - Bishop, AE J1 - Tissue Eng Y1 - 2006/07// VL - 12 SN - 1076-3279 SP - 2025 EP - 2030 N2 - Embryonic stem cells (ESCs) are being investigated increasingly for their potential as a cell source for tissue engineering. Antibiotics are regularly used in ESC culture media to control contamination, although they can be cytotoxic and interfere with protein synthesis. Our aim was to examine the effects of the frequently used antibiotics gentamicin and combined penicillin and streptomycin on ESC culture using differentiation of murine ESC into type II pneumocytes as a model. Antibiotics reduced the expression of the specific marker for type II pneumocytes, SPC mRNA, by up to 60%. We also identified an adverse effect on the growth rate of differentiating embryoid bodies, causing a significant ( p < 0.05) reduction of up to 40%, and an increase in population doubling time of up to 48%. No contamination was seen in any of the cultures. Our findings suggest that the routine use of antibiotics in ESC culture should be avoided as it may reduce the efficiency of the culture system. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16889530&query_hl=1 ER - TY - JFULL T1 - Identification of BAF57 mutations in human breast cancer cell lines. A1 - Kiskinis, E A1 - García-Pedrero, JM A1 - Villaronga, MA A1 - Parker, MG A1 - Belandia, B J1 - Breast Cancer Res Treat Y1 - 2006/07// VL - 98 SN - 0167-6806 SP - 191 EP - 198 N2 - Accumulating genetic and biochemical evidences support a role for the SWI/SNF chromatin-remodeling complex in cancer development and multiple core subunits of these complexes have been found to function as tumor suppressor genes. The core SWI/SNF subunit BAF57 mediates direct interactions with estrogen and androgen receptors (ER and AR) regulating their transcriptional activity. BAF57 gene maps to chromosome band 17 q21 in close proximity to the BRCA1 gene. This locus has been associated with frequent loss of heterozygosity (LOH) and allelic imbalance in breast cancers; however, BRCA1 mutations are rare events in sporadic breast cancer with LOH in the region, suggesting that another tumor suppressor gene resides in this area. All these reasons prompted us to screen for mutations in the BAF57 gene using a panel of the most commonly used human breast cancer cell lines. All cell lines analysed contain wild-type copies of BAF57 gene with the only exception of the breast ductal carcinoma cell line BT549. Sequencing of genomic DNA and cDNA generated from BT549 mRNA demonstrated the presence of a CA dinucleotide insertion in exon 5 of BAF57. The absence of wild-type BAF57 alleles indicates that this is a biallelic inactivating mutation that causes a frameshift and as a consequence a premature stop codon leading to a truncated BAF57 protein. A functional characterisation of the truncated BAF57 showed that it has lost the ability to bind to ER but still binds to the nuclear receptor coactivator SRC1e. Furthermore, we observed that the expression of the truncated BAF57 increased the ability of SRC1e to potentiate transcriptional activation by ERalpha, suggesting that mutations in BAF57 could contribute to the oncogenic transformation in breast cancer cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16538531&query_hl=1 ER - TY - JFULL T1 - Characterization and clinical application of human CD34+ stem/progenitor cell populations mobilized into the blood by granulocyte colony-stimulating factor. A1 - Gordon, MY A1 - Levicar, N A1 - Pai, M A1 - Bachellier, P A1 - Dimarakis, I A1 - Al-Allaf, F A1 - M'Hamdi, H A1 - Thalji, T A1 - Welsh, JP A1 - Marley, SB A1 - Davies, J A1 - Dazzi, F A1 - Marelli-Berg, F A1 - Tait, P A1 - Playford, R A1 - Jiao, L A1 - Jensen, S A1 - Nicholls, JP A1 - Ayav, A A1 - Nohandani, M A1 - Farzaneh, F A1 - Gaken, J A1 - Dodge, R A1 - Alison, M A1 - Apperley, JF A1 - Lechler, R A1 - Habib, NA J1 - Stem Cells Y1 - 2006/07// VL - 24 SN - 1066-5099 SP - 1822 EP - 1830 N2 - A phase I study was performed to determine the safety and tolerability of injecting autologous CD34(+) cells into five patients with liver insufficiency. The study was based on the hypothesis that the CD34(+) cell population in granulocyte colony-stimulating factor (G-CSF)-mobilized blood contains a subpopulation of cells with the potential for regenerating damaged tissue. We separated a candidate CD34(+) stem cell population from the majority of the CD34(+) cells (99%) by adherence to tissue culture plastic. The adherent and nonadherent CD34(+) cells were distinct in morphology, immunophenotype, and gene expression profile. Reverse transcription-polymerase chain reaction-based gene expression analysis indicated that the adherent CD34(+) cells had the potential to express determinants consistent with liver, pancreas, heart, muscle, and nerve cell differentiation as well as hematopoiesis. Overall, the characteristics of the adherent CD34(+) cells identify them as a separate putative stem/progenitor cell population. In culture, they produced a population of cells exhibiting diverse morphologies and expressing genes corresponding to multiple tissue types. Encouraged by this evidence that the CD34(+) cell population contains cells with the potential to form hepatocyte-like cells, we gave G-CSF to five patients with liver insufficiency to mobilize their stem cells for collection by leukapheresis. Between 1 x 10(6) and 2 x 10(8) CD34(+) cells were injected into the portal vein (three patients) or hepatic artery (two patients). No complications or specific side effects related to the procedure were observed. Three of the five patients showed improvement in serum bilirubin and four of five in serum albumin. These observations warrant further clinical trials. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16556705&query_hl=1 ER - TY - JFULL T1 - Hypomorphic promoter mutation in PIGM causes inherited glycosylphosphatidylinositol deficiency. A1 - Almeida, AM A1 - Murakami, Y A1 - Layton, DM A1 - Hillmen, P A1 - Sellick, GS A1 - Maeda, Y A1 - Richards, S A1 - Patterson, S A1 - Kotsianidis, I A1 - Mollica, L A1 - Crawford, DH A1 - Baker, A A1 - Ferguson, M A1 - Roberts, I A1 - Houlston, R A1 - Kinoshita, T A1 - Karadimitris, A J1 - Nat Med Y1 - 2006/07// VL - 12 SN - 1078-8956 SP - 846 EP - 851 N2 - Attachment to the plasma membrane by linkage to a glycosylphosphatidylinositol (GPI) anchor is a mode of protein expression highly conserved from protozoa to mammals. As a clinical entity, deficiency of GPI has been recognized as paroxysmal nocturnal hemoglobinuria, an acquired clonal disorder associated with somatic mutations of the X-linked PIGA gene in hematopoietic cells. We have identified a novel disease characterized by a propensity to venous thrombosis and seizures in which deficiency of GPI is inherited in an autosomal recessive manner. In two unrelated kindreds, a point mutation (c --> g) at position -270 from the start codon of PIGM, a mannosyltransferase-encoding gene, disrupts binding of the transcription factor Sp1 to its cognate promoter motif. This mutation substantially reduces transcription of PIGM and blocks mannosylation of GPI, leading to partial but severe deficiency of GPI. These findings indicate that biosynthesis of GPI is essential to maintain homeostasis of blood coagulation and neurological function. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16767100&query_hl=1 ER - TY - JFULL T1 - The double lysine motif of tapasin is a retrieval signal for retention of unstable MHC class I molecules in the endoplasmic reticulum. A1 - Paulsson, KM A1 - Jevon, M A1 - Wang, JW A1 - Li, S A1 - Wang, P J1 - J Immunol Y1 - 2006/06/15/ VL - 176 SN - 0022-1767 SP - 7482 EP - 7488 N2 - Tapasin (tpn), an essential component of the MHC class I (MHC I) loading complex, has a canonical double lysine motif acting as a retrieval signal, which mediates retrograde transport of escaped endoplasmic reticulum (ER) proteins from the Golgi back to the ER. In this study, we mutated tpn with a substitution of the double lysine motif to double alanine (GFP-tpn-aa). This mutation abolished interaction with the coatomer protein complex I coatomer and resulted in accumulation of GFP-tpn-aa in the Golgi compartment, suggesting that the double lysine is important for the retrograde transport of tpn from late secretory compartments to the ER. In association with the increased Golgi distribution, the amount of MHC I exported from the ER to the surface was increased in 721.220 cells transfected with GFP-tpn-aa. However, the expressed MHC I were less stable and had increased turnover rate. Our results suggest that tpn with intact double lysine retrieval signal regulates retrograde transport of unstable MHC I molecules from the Golgi back to the ER to control the quality of MHC I Ag presentation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16751394&query_hl=1 ER - TY - JFULL T1 - Chondrogenic Differentiation of Human Embryonic Stem Cells: The Effect of the Micro-Environment. A1 - Vats, A A1 - Bielby, RC A1 - Tolley, N A1 - Dickinson, SC A1 - Boccaccini, AR A1 - Hollander, AP A1 - Bishop, AE A1 - Polak, JM J1 - Tissue Eng Y1 - 2006/06/01/ SN - 1076-3279 N2 - We have previously induced differentiation of embryonic stem cells (ESC) to specific phenotypes by manipulating the culture conditions, including the use of indirect co-culture. In this study, we hypothesized that co-culture with primary chondrocytes can induce human embryonic stem cells (hESC) to differentiate towards the chondrocyte lineage. Co-cultures of hESC and chondrocytes were established using well inserts, with control comprising hESC grown alone or with fibroblasts. After 28 days, after removal of the chondrocyte inserts, hESC differentiation was assessed, by morphology, immunocytochemistry, and reverse transcription polymerase chain reaction. hESC, co-cultured or grown alone, were also implanted into SCID mice on a poly-D, L-lactide scaffold, harvested 35 days later and assessed in the same way. hESC co-cultured with chondrocytes formed colonies and secreted extracellular matrix containing glycosaminoglycans (GAG). Quantitative assay showed increased synthesis of sulfated GAG in co-culture as compared with control hESC grown alone for the same period (p < 0.0001). In addition, co-cultured hESC expressed Sox 9 and collagen type II, unlike control hESC. Co-culture with fibroblasts did not induce chondrogenic differentiation. The implanted constructs with co-cultured hESC contained significantly more type II collagen (p < 0.01), type I collagen (p < 0.05), total collagen (p < 0.01), and GAG (p < 0.01) than those with hESC grown alone. Thus, we show for the first time differentiation of hESC to chondrocytes. Our results confirm the potential of the culture micro-environment to influence ESC differentiation and could provide the basis for future generation of chondrogenic cells for use in tissue repair and increase our understanding of the mechanisms that direct differentiation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16796472&query_hl=1 ER - TY - JFULL T1 - Antibiotics Reduce the Growth Rate and Differentiation of Embryonic Stem Cell Cultures. A1 - Cohen, S A1 - Samadikuchaksaraei, A A1 - Polak, JM A1 - Bishop, AE J1 - Tissue Eng Y1 - 2006/06/01/ SN - 1076-3279 N2 - Embryonic stem cells (ESCs) are being investigated increasingly for their potential as a cell source for tissue engineering. Antibiotics are regularly used in ESC culture media to control contamination, although they can be cytotoxic and interfere with protein synthesis. Our aim was to examine the effects of the frequently used antibiotics gentamicin and combined penicillin and streptomycin on ESC culture using differentiation of murine ESC into type II pneumocytes as a model. Antibiotics reduced the expression of the specific marker for type II pneumocytes, SPC mRNA, by up to 60%. We also identified an adverse effect on the growth rate of differentiating embryoid bodies, causing a significant ( p < 0.05) reduction of up to 40%, and an increase in population doubling time of up to 48%. No contamination was seen in any of the cultures. Our findings suggest that the routine use of antibiotics in ESC culture should be avoided as it may reduce the efficiency of the culture system. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16761912&query_hl=1 ER - TY - JFULL T1 - Enhanced derivation of osteogenic cells from murine embryonic stem cells after treatment with HepG2-conditioned medium and modulation of the embryoid body formation period: application to skeletal tissue engineering. A1 - Hwang, YS A1 - Randle, WL A1 - Bielby, RC A1 - Polak, JM A1 - Mantalaris, A J1 - Tissue Eng Y1 - 2006/06// VL - 12 SN - 1076-3279 SP - 1381 EP - 1392 N2 - Despite the considerable progress made in directing embryonic stem cell (ESC) differentiation to therapeutically useful lineages, several issues remain to be resolved before ESCs can be used for cell therapy: 1) increasing the efficiency of specific lineage generation, and 2) developing time- and cost-effective culture systems for controlling ESC differentiation. Our study aimed to develop efficient methods to enhance mesodermal differentiation and thereby upregulate osteogenic differentiation of ESCs. Specifically, murine ESCs (mESCs) were cultured in the presence of 50% conditioned medium (CM) from the human hepatocarcinoma cell line HepG2, which resulted in enhanced mesoderm formation during embryoid body (EB) formation in the CM-treated mESCs (CM-mESCs). By varying the length of EB culture time, we achieved the selective control and stimulation of osteogenic differentiation and suppression of cardiogenic differentiation. Hence, reducing the EB culture of the CM-mESCs to 1 day resulted in 5-10-fold enhancement of osteogenic differentiation, as determined by bone nodule formation, higher alkaline phosphatase activity, the presence of well-organized osteoblast-cadherin in the bone nodules, and increased cbfa-1/runx2 gene expression. In contrast, increasing the EB culture of the CM-mESCs to 5 days resulted in three- to four-fold enhanced cardiogenic differentiation. These findings for development of highly efficient culture systems and protocols for mESC differentiation into osteogenic lineage that are time- and cost-effective can be used in skeletal tissue engineering applications. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16846337&query_hl=1 ER - TY - JFULL T1 - Increase in beta(1)AR-Gi coupling after detubulation in rat ventricular myocytes A1 - Yang, LQ A1 - Gorelik, J A1 - Korchev, Y A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2006/06// VL - 40 SN - 0022-2828 SP - 923 EP - 924 ER - TY - JFULL T1 - Different effects of carvedilol, propranolol and ICI 118 551 on activation of p38 MAP kinase through the beta(2)-adrenoceptor in human myocardium A1 - Zheng, ZL A1 - O'Gara, P A1 - Petrou, M A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2006/06// VL - 40 SN - 0022-2828 SP - 952 EP - 953 ER - TY - JFULL T1 - Chondrogenic differentiation of human embryonic stem cells: the effect of the micro-environment. A1 - Vats, A A1 - Bielby, RC A1 - Tolley, N A1 - Dickinson, SC A1 - Boccaccini, AR A1 - Hollander, AP A1 - Bishop, AE A1 - Polak, JM J1 - Tissue Eng Y1 - 2006/06// VL - 12 SN - 1076-3279 SP - 1687 EP - 1697 N2 - We have previously induced differentiation of embryonic stem cells (ESC) to specific phenotypes by manipulating the culture conditions, including the use of indirect co-culture. In this study, we hypothesized that co-culture with primary chondrocytes can induce human embryonic stem cells (hESC) to differentiate towards the chondrocyte lineage. Co-cultures of hESC and chondrocytes were established using well inserts, with control comprising hESC grown alone or with fibroblasts. After 28 days, after removal of the chondrocyte inserts, hESC differentiation was assessed, by morphology, immunocytochemistry, and reverse transcription polymerase chain reaction. hESC, co-cultured or grown alone, were also implanted into SCID mice on a poly-D, L-lactide scaffold, harvested 35 days later and assessed in the same way. hESC co-cultured with chondrocytes formed colonies and secreted extracellular matrix containing glycosaminoglycans (GAG). Quantitative assay showed increased synthesis of sulfated GAG in co-culture as compared with control hESC grown alone for the same period (p < 0.0001). In addition, co-cultured hESC expressed Sox 9 and collagen type II, unlike control hESC. Co-culture with fibroblasts did not induce chondrogenic differentiation. The implanted constructs with co-cultured hESC contained significantly more type II collagen (p < 0.01), type I collagen (p < 0.05), total collagen (p < 0.01), and GAG (p < 0.01) than those with hESC grown alone. Thus, we show for the first time differentiation of hESC to chondrocytes. Our results confirm the potential of the culture micro-environment to influence ESC differentiation and could provide the basis for future generation of chondrogenic cells for use in tissue repair and increase our understanding of the mechanisms that direct differentiation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16846363&query_hl=1 ER - TY - JFULL T1 - Gram positive bacteria reduce viability and contraction of isolated cardiomyocytes: Role of endothelin-1 A1 - Patel, TA A1 - Mitchell, JA A1 - Warner, TD A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2006/06// VL - 40 SN - 0022-2828 SP - 1007 EP - 1007 ER - TY - JFULL T1 - Effects of phosphodiesterase inhibitors on the contraction of myocytes from rat, guinea pig and human ventricle A1 - Johnson, WB A1 - Katugampola, S A1 - Napier, C A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2006/06// VL - 40 SN - 0022-2828 SP - 954 EP - 954 ER - TY - JFULL T1 - Derivation of distal lung epithelial progenitors from murine embryonic stem cells using a novel three-step differentiation protocol. A1 - Rippon, HJ A1 - Polak, JM A1 - Qin, M A1 - Bishop, AE J1 - Stem Cells Y1 - 2006/05// VL - 24 SN - 1066-5099 SP - 1389 EP - 1398 N2 - Embryonic stem cells (ESCs) are a potential source for the cell-based therapy of a wide variety of lung diseases for which the only current treatment is transplantation. However, distal lung epithelium, like many other endodermally derived somatic cell lineages, is proving difficult to obtain from both murine and human ESCs. We have previously obtained alveolar epithelium from ESCs, although final cell yield remained extremely low. Here, we present an optimized three-step protocol for the derivation of distal lung epithelial cells from murine ESCs. This protocol incorporates (a) treatment of early differentiating embryoid bodies with activin A to enhance the specification of the endodermal germ layer, followed by (b) adherent culture in serum-free medium and (c) the final application of a commercial, lung-specific medium. As well as enhancing the specification of distal lung epithelium, this protocol was found to yield cells with a phenotype most closely resembling that of lung-committed progenitor cells present in the foregut endoderm and the early lung buds during embryonic development. This is in contrast to our previous differentiation method, which drives differentiation through to mature type II alveolar epithelial cells. The derivation of a committed lung progenitor cell type from ESCs is particularly significant for regenerative medicine because the therapeutic implantation of progenitor cells has several clear advantages over the transplantation of mature, terminally differentiated somatic cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16456134&query_hl=1 ER - TY - JFULL T1 - Chromatin signatures of pluripotent cell lines. A1 - Azuara, V A1 - Perry, P A1 - Sauer, S A1 - Spivakov, M A1 - Jørgensen, HF A1 - John, RM A1 - Gouti, M A1 - Casanova, M A1 - Warnes, G A1 - Merkenschlager, M A1 - Fisher, AG J1 - Nat Cell Biol Y1 - 2006/05// VL - 8 SN - 1465-7392 SP - 532 EP - 538 N2 - Epigenetic genome modifications are thought to be important for specifying the lineage and developmental stage of cells within a multicellular organism. Here, we show that the epigenetic profile of pluripotent embryonic stem cells (ES) is distinct from that of embryonic carcinoma cells, haematopoietic stem cells (HSC) and their differentiated progeny. Silent, lineage-specific genes replicated earlier in pluripotent cells than in tissue-specific stem cells or differentiated cells and had unexpectedly high levels of acetylated H3K9 and methylated H3K4. Unusually, in ES cells these markers of open chromatin were also combined with H3K27 trimethylation at some non-expressed genes. Thus, pluripotency of ES cells is characterized by a specific epigenetic profile where lineage-specific genes may be accessible but, if so, carry repressive H3K27 trimethylation modifications. H3K27 methylation is functionally important for preventing expression of these genes in ES cells as premature expression occurs in embryonic ectoderm development (Eed)-deficient ES cells. Our data suggest that lineage-specific genes are primed for expression in ES cells but are held in check by opposing chromatin modifications. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16570078&query_hl=1 ER - TY - JFULL T1 - Clinical applications of stem cell therapy - the pros and cons of stem cell sources A1 - Habib, NA A1 - Gordon, MY J1 - REGEN MED Y1 - 2006/05// VL - 1 SP - 301 EP - 302 ER - TY - JFULL T1 - Acute liver failure: a review. A1 - Khan, SA A1 - Shah, N A1 - Williams, R A1 - Jalan, R J1 - Clin Liver Dis Y1 - 2006/05// VL - 10 SN - 1089-3261 N2 - Since publication of the first descriptions of acute liver failure (ALF) as a distinct clinical entity in the 1950's, the understanding of the pathophysiologic mechanisms involved and the management options have increased substantially. ALF still represents a major challenge for todays hepatologists, because it can rapidly lead to multiorgan failure and death that may be preventable with appropriate intervention. This article summarizes the basic patho-physiology underlying ALF, compares epidemiologic trends in the United States, the United Kingdom, and the Far East, and reviews prognostic markers and treatment options for ALF. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16971260&query_hl=1 ER - TY - JFULL T1 - Analysis of p53 mutations for a mutational signature in human intrahepatic cholangiocarcinoma. A1 - Khan, SA A1 - Taylor-Robinson, SD A1 - Carmichael, PL A1 - Habib, N A1 - Lemoine, NR A1 - Thomas, HC J1 - Int J Oncol Y1 - 2006/05// VL - 28 SN - 1019-6439 SP - 1269 EP - 1277 N2 - Cholangiocarcinoma development may be related to cholangiocyte DNA damage from genotoxic compounds in bile. We have previously shown that human biliary tissue is exposed to genotoxic agents, as evidenced by the presence of DNA adducts. Establishing the presence of a 'mutational signature' in tumour suppressor genes from tumour tissue provides a means of linking cause and effect in human cancer. Inactivation of p53, known to have 'hot-spots' for particular chemical carcinogens, has previously been linked to human cholangiocarcinoma. However, previous p53 studies have focused on exons 5-8, potentially missing gene alterations at other sites. This study examined the putative link between environmental carcinogens and intrahepatic cholangiocarcinoma by analysing DNA from 31 patients for complete p53 mutational signatures, using single strand conformational polymorphism and polymerase chain reaction. All mutations found were compared to known p53 mutations in cholangiocarcinoma and to mutations induced by environmental mutagens, as described in p53 databases. Five non-silent p53 mutations were found, including three new frameshift mutations and two new intron mutations which have not previously been reported in cholangiocarcinoma. Two frameshifts were due to deletions and the third due to an insertion in exon 5. There was no predominant mutational spectrum amongst the set of cholangiocarcinoma samples studied, or on combining these mutations with the dataset of known p53 mutations in cholangiocarcinoma. Several reasons may explain this, including lack of data outside exons 5-8, bias in mutation reporting, the involvement of mutations in non-coding regions or genes other than p53, or the possibility that there is no carcinogenic specific agent and therefore no signature. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16596244&query_hl=1 ER - TY - JFULL T1 - Statins modulate the growth of human pulmonary smooth muscle cells through the mevalonate pathway A1 - Ali, O A1 - Growcott, E A1 - Wharton, J A1 - Wilkins, M J1 - HEART Y1 - 2006/05// VL - 92 SN - 1355-6037 SP - A14 EP - A15 ER - TY - JFULL T1 - Characterization of human fetal osteoblasts by microarray analysis following stimulation with 58S bioactive gel-glass ionic dissolution products. A1 - Christodoulou, I A1 - Buttery, LD A1 - Tai, G A1 - Hench, LL A1 - Polak, JM J1 - J Biomed Mater Res B Appl Biomater Y1 - 2006/05// VL - 77 SN - 1552-4973 SP - 431 EP - 446 N2 - Bioactive glasses dissolve upon immersion in culture medium, releasing their constitutive ions in solution. There is evidence suggesting that these ionic dissolution products influence osteoblast-specific processes. Here, we investigated the effect of 58S sol-gel-derived bioactive glass (60 mol % SiO2, 36 mol % CaO, 4 mol % P2O5) dissolution products on primary osteoblasts derived from human fetal long bone explant cultures (hFOBs). We used U133A human genome GeneChip oligonucleotide arrays to examine 22,283 transcripts and variants, which represent over 18,000 well-substantiated human genes. Hybridization of samples (biotinylated cRNA) derived from monolayer cultures of hFOBs on the arrays revealed that 10,571 transcripts were expressed by these cells, with high confidence. These included transcripts representing osteoblast-related genes coding for growth factors and their associated molecules or receptors, protein components of the extracellular matrix (ECM), enzymes involved in degradation of the ECM, transcription factors, and other important osteoblast-associated markers. A 24-h treatment with a single dosage of ionic products of sol-gel 58S dissolution induced the differential expression of a number of genes, including IL-6 signal transducer/gp130, ISGF-3/STAT1, HIF-1 responsive RTP801, ERK1 p44 MAPK (MAPK3), MAPKAPK2, IGF-I and IGFBP-5. The over 2-fold up-regulation of gp130 and MAPK3 and down-regulation of IGF-I were confirmed by real-time RT-PCR analysis. These data suggest that 58S ionic dissolution products possibly mediate the bioactive effect of 58S through components of the IGF system and MAPK signaling pathways. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16333845&query_hl=1 ER - TY - JFULL T1 - The effects of atorvastatin on long-term prognosis in patients with chronic heart failure. A1 - Xie, RQ A1 - Cui, W A1 - Liu, F A1 - Hao, YM A1 - Zu, XG A1 - Du, J J1 - AM J CARDIOL Y1 - 2006/04/26/ VL - 97 SN - 0002-9149 SP - 60D EP - 61D ER - TY - JFULL T1 - Remote preconditioning induced by skeletal muscle ischemia reducing matrix metalloproteinase expression in porcine myocardium. A1 - Xie, RQ A1 - Cui, W A1 - Liu, F A1 - Hao, YM A1 - Du, J A1 - Li, BH J1 - AM J CARDIOL Y1 - 2006/04/26/ VL - 97 SN - 0002-9149 SP - 55D EP - 55D ER - TY - JFULL T1 - The performance of single Judkins left catheter for transradial coronary angiography: Randomized comparison with brachial type catheter. A1 - Cui, W A1 - Liu, F A1 - Xie, RQ A1 - Yang, XH A1 - Gu, GQ A1 - Lu, JC A1 - Yang, XC A1 - Du, J J1 - AM J CARDIOL Y1 - 2006/04/26/ VL - 97 SN - 0002-9149 SP - 16D EP - 16D ER - TY - JFULL T1 - The role of bradykinin and opioid receptors in remote preconditioning induced by skeletal muscle ischemia. A1 - Xie, RQ A1 - Cui, W A1 - Hao, YM A1 - Liu, F A1 - Du, J A1 - Li, BH J1 - AM J CARDIOL Y1 - 2006/04/26/ VL - 97 SN - 0002-9149 SP - 89D EP - 89D ER - TY - JFULL T1 - Regulation of hematopoiesis in vitro and in vivo by invariant NKT cells. A1 - Kotsianidis, I A1 - Silk, JD A1 - Spanoudakis, E A1 - Patterson, S A1 - Almeida, A A1 - Schmidt, RR A1 - Tsatalas, C A1 - Bourikas, G A1 - Cerundolo, V A1 - Roberts, IA A1 - Karadimitris, A J1 - Blood Y1 - 2006/04/15/ VL - 107 SN - 0006-4971 SP - 3138 EP - 3144 N2 - Invariant natural killer T cells (iNKT cells) are a small subset of immunoregulatory T cells highly conserved in humans and mice. On activation by glycolipids presented by the MHC-like molecule CD1d, iNKT cells promptly secrete T helper 1 and 2 (Th1/2) cytokines but also cytokines with hematopoietic potential such as GM-CSF. Here, we show that the myeloid clonogenic potential of human hematopoietic progenitors is increased in the presence of glycolipid-activated, GM-CSF-secreting NKT cells; conversely, short- and long-term progenitor activity is decreased in the absence of NKT cells, implying regulation of hematopoiesis in both the presence and the absence of immune activation. In accordance with these findings, iNKT-cell-deficient mice display impaired hematopoiesis characterized by peripheral-blood cytopenias, reduced marrow cellularity, lower frequency of hematopoietic stem cells (HSCs), and reduced early and late hematopoietic progenitors. We also show that CD1d is expressed on human HSCs. CD1d-expressing HSCs display short- and long-term clonogenic potential and can present the glycolipid alpha-galactosylceramide to iNKT cells. Thus, iNKT cells emerge as the first subset of regulatory T cells that are required for effective hematopoiesis in both steady-state conditions and under conditions of immune activation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16373666&query_hl=1 ER - TY - JFULL T1 - Derivation of distal airway epithelium from human embryonic stem cells. A1 - Samadikuchaksaraei, A A1 - Cohen, S A1 - Isaac, K A1 - Rippon, HJ A1 - Polak, JM A1 - Bielby, RC A1 - Bishop, AE J1 - Tissue Eng Y1 - 2006/04// VL - 12 SN - 1076-3279 SP - 867 EP - 875 N2 - The pluripotency of embryonic stem cells (ESC) is offering new opportunities in tissue engineering and cell therapy. We have shown previously that alveolar epithelial cells, specifically type II pneumocytes, can be derived from murine ESC and hypothesized that a similar protocol could be used successfully on human ESC. Undifferentiated human ESC were induced to form embryoid bodies that were transferred into adherent culture conditions and grown in a medium designed for the maintenance of mature small airway epithelium. On inverted microscopy, the generated cells showed the cobblestone-like morphology of epithelium. The presence of surfactant protein C, a specific marker of type II pneumocytes, and its corresponding RNA were demonstrated by immunostaining and reverse transcription polymerase chain reaction, respectively. Electron microscopy revealed frequent cells with the typical ultrastructure of type II pneumocytes. This study provides evidence for in vitro induction of the differentiation from human ESC of alveolar type II cells, which have the potential for therapeutic use or construction of an in vitro model of human lung. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16674299&query_hl=1 ER - TY - JFULL T1 - Cell extract-based re-programming of embryonic stem cells to promote lineage-specific differentiation A1 - Qin, MD A1 - Tai, GP A1 - Collas, P A1 - Polak, JM A1 - Bishop, AE J1 - TISSUE ENG Y1 - 2006/04// VL - 12 SN - 1076-3279 SP - 1050 EP - 1051 ER - TY - JFULL T1 - Stem cells and tissue engineering: past, present, and future. A1 - Polak, JM A1 - Bishop, AE J1 - Ann N Y Acad Sci Y1 - 2006/04// VL - 1068 SN - 0077-8923 SP - 352 EP - 366 N2 - Tissue engineering is an interdisciplinary field that brings together the principles of the life sciences and medicine with those of engineering. The increase in its development over the past decade has resulted from a variety of factors; advances in genomics and proteomics, the advent of new biomaterials as potential templates for tissue growth, improvements in bioreactor design, and increased understanding of healing processes. Possibly the greatest contribution has come from our increased knowledge and understanding of stem cell biology, which is paving the way for the generation of unlimited cells of specific phenotypes for incorporation into engineered tissue constructs. Thus, tissue engineering approaches for expanding and engrafting the differentiated progeny of embryonic, fetal, or adult stem cells have major potential for tissue repair and will make a major contribution to medicine in the 21st century. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16831937&query_hl=1 ER - TY - JFULL T1 - Evaluation of frequency, type, and function of gap junctions between skeletal myoblasts overexpressing connexin43 and cardiomyocytes: relevance to cell transplantation. A1 - Stagg, MA A1 - Coppen, SR A1 - Suzuki, K A1 - Varela-Carver, A A1 - Lee, J A1 - Brand, NJ A1 - Fukushima, S A1 - Yacoub, MH A1 - Terracciano, CM J1 - FASEB J Y1 - 2006/04// VL - 20 SN - 1530-6860 SP - 744 EP - 746 N2 - Cell transplantation of skeletal myoblasts (SMs) is one possible treatment for repairing cardiac tissue after myocardial injury. However, inappropriate electrical coupling between grafted SMs and host cardiomyocytes may be responsible for the arrhythmias observed in clinical trials of SM transplantation. Whether functional gap junctions occur between the two cell types remains controversial. We have studied the ability of SMs to electrically couple with isolated adult rat cardiomyocytes (CMs) and assessed whether connexin43 (Cx43) overexpression enhanced gap junctional conductance (Gj). C2C12 myoblast lines overexpressing Cx43 were generated by gene transfection and clonal selection. CMs were cocultured with either SMs overexpressing Cx43 (CM-SM(Cx43)) or control SMs (CM-SM(WT)) in vitro. Gj between pairs of SMs and CMs was quantified with dual whole cell patch clamping. Formation of Gj occurred between 22% of CM-SM(WT) pairs (n=73) and 48% of CM-SM(Cx43) pairs (n=71, P<0.001). The Gj of CM-SM(Cx43) pairs (29.7+/-4.3 nS, n=21) was greater than that of CM-SM(WT) pairs (14.8+/-2.0 nS, n=12, P<0.05). The overexpression of Cx43 in SMs increased the formation of electrical communication and the steady-state conductance between SMs and CMs. Enhanced gap junctional conductance may be useful to promote the integration of transplanted SMs into the myocardium. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16443678&query_hl=1 ER - TY - JFULL T1 - Functional characterization of embryonic stem cell-derived cardiomyocytes using scanning ion conductance microscopy. A1 - Gorelik, J A1 - Ali, NN A1 - Shevchuk, AI A1 - Lab, M A1 - Williamson, C A1 - Harding, SE A1 - Korchev, YE J1 - Tissue Eng Y1 - 2006/04// VL - 12 SN - 1076-3279 SP - 657 EP - 664 N2 - We report here the novel use of scanning ion conductance microscopy (SICM) to produce surface images of embryonic stem cell-derived cardiomyocytes (ESCM) to identify individual contracting cardiomyocytes among different cell types. By measuring amplitude and rhythm we can quantitate contraction of ESCM. This method gives, within the same experiment, an assessment of the number and position of ESCM within the layer of mixed cell types, as well as an accurate measure of the response of individual ESCM. Using different modulators of contraction as examples we showed how SICM could be used for recording their responses. We subsequently demonstrated that this model can be used to investigate the protective effect of antiarrhythmogenic drugs. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16674281&query_hl=1 ER - TY - JFULL T1 - Characterisation of the biochemical and genetic defect in congenital glycosylphosphatidylinositol (GPI) deficiency A1 - Almeida, A A1 - Murakami, Y A1 - Hillmen, P A1 - Richards, SJ A1 - Maeda, Y A1 - Sellick, GS A1 - Crawford, DH A1 - Baker, A A1 - Ferguson, MA A1 - Houslton, R A1 - Roberts, IA A1 - Kinoshita, T A1 - Layton, DM A1 - Karadimitris, A J1 - BRIT J HAEMATOL Y1 - 2006/04// VL - 133 SN - 0007-1048 SP - 23 EP - 23 ER - TY - JFULL T1 - Cloning and expression of rat liver S-adenosylmethionine synthetase A1 - Cui, W A1 - Huang, L A1 - Li, W A1 - Wang, LP J1 - CHEM RES CHINESE U Y1 - 2006/03// VL - 22 SN - 1005-9040 SP - 236 EP - 238 N2 - The S-adenosylmethionine synthetase (SAM synthetase) is responsible for in vivo synthesis of S-adenosylmethionine(SAM), which is a kind of biologically active molecules distributed in all body tissues and fluids and involved in a number of biochemical reactions. In this study, a cDNA containing the coding sequence for rat liver,SAM synthetase was cloned into the prokaryotic expression vector pQE30 and expressed in E. coli M15. A major band corresponding to a protein of 48 kDa was detected on SDS-PAGE. The protein was distributed in both the soluble fraction and the insoluble fraction. In soluble fractions the protein was fully active. ER - TY - JFULL T1 - Inflammation and sex steroid receptors: a motif for change. A1 - Brosens, JJ A1 - Lam, EW A1 - Parker, MG J1 - Cell Y1 - 2006/02/10/ VL - 124 SN - 0092-8674 SP - 466 EP - 468 N2 - Homeostasis in reproductive tissues requires integration of hormonal and inflammatory signals. In this issue of Cell, Zhu et al. (2006) discover that proinflammatory signals switch repressed steroid hormone receptors into transcriptional activators by targeting TAB2, an adaptor protein that tethers corepressors. These findings have implications for the treatment of endocrine-resistant cancers. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16469693&query_hl=1 ER - TY - JFULL T1 - Matrix metalloproteinase (MMP) and tissue inhibitor of MMPs (TIMP) gene expression in myocardial recovery from heart failure A1 - Felkin, LE A1 - Birks, EJ A1 - Soppa, GK A1 - Terracciano, CM A1 - Yacoub, MH A1 - Barton, PJR J1 - HEART Y1 - 2006/02// VL - 92 SN - 1355-6037 ER - TY - JFULL T1 - No evidence of mutations of the PSMB5 (beta-5 subunit of proteasome) in a case of myeloma with clinical resistance to Bortezomib. A1 - Politou, M A1 - Karadimitris, A A1 - Terpos, E A1 - Kotsianidis, I A1 - Apperley, JF A1 - Rahemtulla, A J1 - Leuk Res Y1 - 2006/02// VL - 30 SN - 0145-2126 SP - 240 EP - 241 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16081156&query_hl=1 ER - TY - JFULL T1 - Upregulated genes in sporadic, idiopathic pulmonary arterial hyptension. A1 - Edgar A A1 - Chacon MR A1 - Bishop AE A1 - Yacoub MH J1 - Respiratory Research Y1 - 2006/01/03/ VL - 7 SP - 1 EP - 14 ER - TY - JFULL T1 - N-acetylaspartate metabolism in neural cells. A1 - Bhakoo, KK A1 - Craig, T A1 - Pearce, D J1 - Adv Exp Med Biol Y1 - 2006/// VL - 576 SN - 0065-2598 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16802703&query_hl=1 ER - TY - JFULL T1 - Neural induction promotes large-scale chromatin reorganisation of the Mash1 locus. A1 - Williams, RR A1 - Azuara, V A1 - Perry, P A1 - Sauer, S A1 - Dvorkina, M A1 - Jørgensen, H A1 - Roix, J A1 - McQueen, P A1 - Misteli, T A1 - Merkenschlager, M A1 - Fisher, AG J1 - J Cell Sci Y1 - 2006/01/01/ VL - 119 SN - 0021-9533 SP - 132 EP - 140 N2 - Determining how genes are epigenetically regulated to ensure their correct spatial and temporal expression during development is key to our understanding of cell lineage commitment. Here we examined epigenetic changes at an important proneural regulator gene Mash1 (Ascl1), as embryonic stem (ES) cells commit to the neural lineage. In ES cells where the Mash1 gene is transcriptionally repressed, the locus replicated late in S phase and was preferentially positioned at the nuclear periphery with other late-replicating genes (Neurod, Sprr2a). This peripheral location was coupled with low levels of histone H3K9 acetylation at the Mash1 promoter and enhanced H3K27 methylation but surprisingly location was not affected by removal of the Ezh2/Eed HMTase complex or several other chromatin-silencing candidates (G9a, SuV39h-1, Dnmt-1, Dnmt-3a and Dnmt-3b). Upon neural induction however, Mash1 transcription was upregulated (>100-fold), switched its time of replication from late to early in S phase and relocated towards the interior of the nucleus. This spatial repositioning was selective for neural commitment because Mash1 was peripheral in ES-derived mesoderm and other non-neural cell types. A bidirectional analysis of replication timing across a 2 Mb region flanking the Mash1 locus showed that chromatin changes were focused at Mash1. These results suggest that Mash1 is regulated by changes in chromatin structure and location and implicate the nuclear periphery as an important environment for maintaining the undifferentiated state of ES cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16371653&query_hl=1 ER - TY - JFULL T1 - Derivation and characterization of alveolar epithelial cells from murine embryonic stem cells in vitro. A1 - Samadikuchaksaraei, A A1 - Bishop, AE J1 - Methods Mol Biol Y1 - 2006/// VL - 330 SN - 1064-3745 SP - 233 EP - 248 N2 - We present a protocol that has been developed for induction of the differentiation of murine embryonic stem (ES) cells to alveolar type II cells. With this protocol, undifferentiated murine ES cells are induced to form embryoid bodies (EBs). The 10-d-old EBs are transferred to adherent culture conditions and are fed with high-glucose Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% v/v fetal bovine serum, 2 mM L-glutamine, and 0.1 mM 2-mercaptoethanol for 20 d without splitting. Then, the cells are fed with a medium designed for the maintenance and growth of mature distal airway epithelial cells (small airway growth medium, SAGM) for 3 d. Characterization of the alveolar type II cells was done using real-time reverse transcriptase polymerase chain reaction detection of surfactant protein C mRNA and immunocytochemical detection of prosurfactant protein C. Real-time reverse transcriptase polymerase chain reaction revealed that SAGM increases the mRNA expression level of SPC by a factor of 8 when compared to that of cells grown in supplemented high-glucose DMEM (p < 0.05, Student t-test). Immunocytochemistry revealed that proSPC-expressing cells comprised 2.8 +/- 0.23% of the total cell population in SAGM-treated samples and 0.5 +/- 0.1% in samples treated with supplemented high-glucose DMEM (p < 0.05, chi2 test). L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16846028&query_hl=1 ER - TY - JFULL T1 - Nuclear receptor regulation gears up another Notch. A1 - Belandia, B A1 - Parker, MG J1 - Nucl Recept Signal Y1 - 2006/// VL - 4 SN - 1550-7629 SP - e001 EP - e001 N2 - In this perspective we describe examples of crosstalk between nuclear receptors (NRs) and Notch signaling by means of direct functional interactions between components of both pathways. This crosstalk may provide eukaryotic organisms with molecular mechanisms for the coordination of llong-distance endocrine signals with cell-to-cell juxtacrine communication. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16604164&query_hl=1 ER - TY - JFULL T1 - Phosphodiesterase type 4 expression and anti-proliferative effects in human pulmonary artery smooth muscle cells. A1 - Growcott, EJ A1 - Spink, KG A1 - Ren, X A1 - Afzal, S A1 - Banner, KH A1 - Wharton, J J1 - Respir Res Y1 - 2006/// VL - 7 SN - 1465-993X SP - 9 EP - 9 N2 - BACKGROUND: Pulmonary arterial hypertension is a proliferative vascular disease, characterized by aberrant regulation of smooth muscle cell proliferation and apoptosis in distal pulmonary arteries. Prostacyclin (PGI2) analogues have anti-proliferative effects on distal human pulmonary artery smooth muscle cells (PASMCs), which are dependent on intracellular cAMP stimulation. We therefore sought to investigate the involvement of the main cAMP-specific enzymes, phosphodiesterase type 4 (PDE4), responsible for cAMP hydrolysis. METHODS: Distal human PASMCs were derived from pulmonary arteries by explant culture (n = 14, passage 3-12). Responses to platelet-derived growth factor-BB (5-10 ng/ml), serum, PGI2 analogues (cicaprost, iloprost) and PDE4 inhibitors (roflumilast, rolipram, cilomilast) were determined by measuring cAMP phosphodiesterase activity, intracellular cAMP levels, DNA synthesis, apoptosis (as measured by DNA fragmentation and nuclear condensation) and matrix metalloproteinase-2 and -9 (MMP-2, MMP-9) production. RESULTS: Expression of all four PDE4A-D genes was detected in PASMC isolates. PDE4 contributed to the main proportion (35.9 +/- 2.3%, n = 5) of cAMP-specific hydrolytic activity demonstrated in PASMCs, compared to PDE3 (21.5 +/- 2.5%), PDE2 (15.8 +/- 3.4%) or PDE1 activity (14.5 +/- 4.2%). Intracellular cAMP levels were increased by PGI2 analogues and further elevated in cells co-treated with roflumilast, rolipram and cilomilast. DNA synthesis was attenuated by 1 microM roflumilast (49 +/- 6% inhibition), rolipram (37 +/- 6%) and cilomilast (30 +/- 4%) and, in the presence of 5 nM cicaprost, these compounds exhibited EC50 values of 4.4 (2.6-6.1) nM (Mean and 95% confidence interval), 59 (36-83) nM and 97 (66-130) nM respectively. Roflumilast attenuated cell proliferation and gelatinase (MMP-2 and MMP-9) production and promoted the anti-proliferative effects of PGI2 analogues. The cAMP activators iloprost and forskolin also induced apoptosis, whereas roflumilast had no significant effect. CONCLUSION: PDE4 enzymes are expressed in distal human PASMCs and the effects of cAMP-stimulating agents on DNA synthesis, proliferation and MMP production is dependent, at least in part, on PDE4 activity. PDE4 inhibition may provide greater control of cAMP-mediated anti-proliferative effects in human PASMCs and therefore could prove useful as an additional therapy for pulmonary arterial hypertension. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16423283&query_hl=1 ER - TY - JFULL T1 - Contribution of three-dimensional trabecular bone microstructure of the proximal femur to its mechanical properties as assessed by micro-finite element analysis A1 - Cui, W A1 - Won, YY A1 - Baek, MH A1 - Kim, K J1 - KEY ENG MAT Y1 - 2006/// VL - 321-323 SP - 278 EP - 281 N2 - The purpose of this study was to investigate the contribution of the microstructural properties of trabecular bone in predicting its elastic modulus in the intertrochanteric region. A total of 15 trabecular bone core specimens were obtained from the proximal femurs of patients undergoing total hip arthroplasty. The micro-computed tomography (micro-CT) was used to scan each specimen to obtain micro-morphology. Microstructural parameters were directly calculated using software. Micro-CT images were converted to micro-finite element model using meshing technique, and then micro-finite element analysis (FEA) was performed to assess the mechanical property (Young's modulus) of trabecular bone. The results showed that the ability to explain this variance of Young's modulus is improved by combining the structural indices with each other. It suggested that assessment of bone microarchitecture should be added as regards detection of osteoporosis and evaluation of the efficacy of drug treatments for osteoporosis. ER - TY - JFULL T1 - Profiling of DNA replication timing in unsynchronized cell populations. A1 - Azuara, V J1 - Nat Protoc Y1 - 2006/// VL - 1 SN - 1750-2799 SP - 2171 EP - 2177 N2 - Profiling chromatin in a particular cell type provides a valuable 'signature' for cell identity and developmental stage. One approach has been to assay and use the timing of DNA replication across a panel of loci as an indicator of chromatin structure. This epigenetic profiling used on pluripotent embryonic stem (ES) cells has reliably distinguished them from cells that have a more restricted lineage potential. Thus, such an approach may become increasingly useful for understanding the molecular basis of pluripotency and lineage induction, especially in the context of stem-cell therapy. Here I describe in detail the DNA replication timing method, whereby unsynchronized cell populations are pulse-labeled with 5-bromo-2'-deoxyuridine (BrdU), fractionated according to cell-cycle stage and the abundance of candidate sequences within newly replicated DNA is determined by PCR. This robust protocol has been used consistently by several laboratories and might offer some advantages over conventional transcription-based profiling for characterizing cell populations. The procedure requires 3-4 d to complete. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17487209&query_hl=1 ER - TY - JFULL T1 - Pulmonary epithelium. A1 - Bishop, AE A1 - Polak, JM J1 - Methods Enzymol Y1 - 2006/// VL - 418 SN - 0076-6879 SP - 333 EP - 349 N2 - Repair or regeneration of defective lung epithelium would be of great therapeutic potential. Cellular sources for such repair have long been searched for within the lung, but the identification and characterization of stem or progenitor cells have been hampered by the complexity and cellular heterogeneity of the organ. In recent years, various pulmonary cells have been identified that meet the criteria for stem cells but it remains to be seen how far manipulation of these tissue-specific cell pools can upregulate epithelial repair. The initial excitement that greeted the results of animal experiments showing cells of bone marrow origin in murine lung has been tempered by more recent data suggesting that the cells do not repair pulmonary epithelium. However, there are reports of engraftment of bone marrow-derived cells in human lung, albeit at a low level, so the administration of cell therapy via the circulation, for repair and/or gene delivery, needs further investigation. The potential of human embryonic stem cells to generate any cell, tissue, or organ on demand for tissue repair or replacement is promising to revolutionize the treatment of human disease. Although some headway has been made into making pulmonary epithelium from these stem cells, human embryonic stem cell technology is still in its infancy and many technical, safety, and ethical hurdles must be cleared before clinical trials can begin. This chapter focuses on the potential role of stem cells in future approaches to lung repair and regeneration. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17141045&query_hl=1 ER - TY - JFULL T1 - Preservation of rat aortic tissue transplant with green tea polyphenols. A1 - Hyon, SH A1 - Kim, DH A1 - Cui, W A1 - Matsumura, K A1 - Kim, JY A1 - Tsutsumi, S J1 - Cell Transplant Y1 - 2006/// VL - 15 SN - 0963-6897 SP - 881 EP - 883 N2 - Green tea polyphenols have recently attracted medical attention as bioactive agents with anticancer, antimicrobial, and antiviral effects. We discovered their new usage as preservative agents for tissue transplants. We preserved rat aortas in a DMEM solution containing polyphenols extracted from green tea leaves. The preserved aortas retained original structures and mechanical strength, and were devoid of any undesirable cell secretions for over a month under physiological conditions. In addition, aortas from Lewis rats preserved for a month and transplanted to allogenic ACI rats completely avoided rejection by the host, suggesting that the polyphenols have immunosuppressive actions on the aortic tissues. From these results, we conclude that polyphenol treatment of aortic tissue transplant can maintain its viability for extended periods of time either before or after transplantation, and the method can be applicable to other transplantation situations. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17299992&query_hl=1 ER - TY - JFULL T1 - ZNF366 is an estrogen receptor corepressor that acts through CtBP and histone deacetylases. A1 - Lopez-Garcia, J A1 - Periyasamy, M A1 - Thomas, RS A1 - Christian, M A1 - Leao, M A1 - Jat, P A1 - Kindle, KB A1 - Heery, DM A1 - Parker, MG A1 - Buluwela, L A1 - Kamalati, T A1 - Ali, S J1 - Nucleic Acids Res Y1 - 2006/// VL - 34 SN - 1362-4962 SP - 6126 EP - 6136 N2 - The regulation of gene expression by estrogen receptor-alpha (ERalpha) requires the coordinated and temporal recruitment of diverse sets of transcriptional co-regulator complexes, which mediate nucleosome remodelling and histone modification. Using ERalpha as bait in a yeast two-hybrid screen, we have identified a novel ERalpha-interacting protein, ZNF366, which is a potent corepressor of ERalpha activity. The interaction between ZNF366 and ERalpha has been confirmed in vitro and in vivo, and is mediated by the zinc finger domains of the two proteins. Further, we show that ZNF366 acts as a corepressor by interacting with other known ERalpha corepressors, namely RIP140 and CtBP, to inhibit expression of estrogen-responsive genes in vivo. Together, our results indicate that ZNF366 may play an important role in regulating the expression of genes in response to estrogen. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17085477&query_hl=1 ER - TY - JFULL T1 - Upregulated genes in sporadic, idiopathic pulmonary arterial hypertension. A1 - Edgar, AJ A1 - Chacón, MR A1 - Bishop, AE A1 - Yacoub, MH A1 - Polak, JM J1 - Respir Res Y1 - 2006/// VL - 7 SN - 1465-993X SP - 1 EP - 1 N2 - BACKGROUND: To elucidate further the pathogenesis of sporadic, idiopathic pulmonary arterial hypertension (IPAH) and identify potential therapeutic avenues, differential gene expression in IPAH was examined by suppression subtractive hybridisation (SSH). METHODS: Peripheral lung samples were obtained immediately after removal from patients undergoing lung transplant for IPAH without familial disease, and control tissues consisted of similarly sampled pieces of donor lungs not utilised during transplantation. Pools of lung mRNA from IPAH cases containing plexiform lesions and normal donor lungs were used to generate the tester and driver cDNA libraries, respectively. A subtracted IPAH cDNA library was made by SSH. Clones isolated from this subtracted library were examined for up regulated expression in IPAH using dot blot arrays of positive colony PCR products using both pooled cDNA libraries as probes. Clones verified as being upregulated were sequenced. For two genes the increase in expression was verified by northern blotting and data analysed using Student's unpaired two-tailed t-test. RESULTS: We present preliminary findings concerning candidate genes upregulated in IPAH. Twenty-seven upregulated genes were identified out of 192 clones examined. Upregulation in individual cases of IPAH was shown by northern blot for tissue inhibitor of metalloproteinase-3 and decorin (P < 0.01) compared with the housekeeping gene glyceraldehydes-3-phosphate dehydrogenase. CONCLUSION: Four of the up regulated genes, magic roundabout, hevin, thrombomodulin and sucrose non-fermenting protein-related kinase-1 are expressed specifically by endothelial cells and one, muscleblind-1, by muscle cells, suggesting that they may be associated with plexiform lesions and hypertrophic arterial wall remodelling, respectively. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16390543&query_hl=1 ER - TY - JFULL T1 - Suppression of oxidative metabolism and mitochondrial biogenesis by the transcriptional corepressor RIP140 in mouse adipocytes. A1 - Powelka, AM A1 - Seth, A A1 - Virbasius, JV A1 - Kiskinis, E A1 - Nicoloro, SM A1 - Guilherme, A A1 - Tang, X A1 - Straubhaar, J A1 - Cherniack, AD A1 - Parker, MG A1 - Czech, MP J1 - J Clin Invest Y1 - 2006/01// VL - 116 SN - 0021-9738 SP - 125 EP - 136 N2 - Using an siRNA-based screen, we identified the transcriptional corepressor RIP140 as a negative regulator of insulin-responsive hexose uptake and oxidative metabolism in 3T3-L1 adipocytes. Affymetrix GeneChip profiling revealed that RIP140 depletion upregulates the expression of clusters of genes in the pathways of glucose uptake, glycolysis, TCA cycle, fatty acid oxidation, mitochondrial biogenesis, and oxidative phosphorylation in these cells. Conversely, we show that reexpression of RIP140 in mouse embryonic fibroblasts derived from RIP140-null mice downregulates expression of many of these same genes. Consistent with these microarray data, RIP140 gene silencing in cultured adipocytes increased both conversion of [14C]glucose to CO2 and mitochondrial oxygen consumption. RIP140-null mice, previously reported to resist weight gain on a high-fat diet, are shown here to display enhanced glucose tolerance and enhanced responsiveness to insulin compared with matched wild-type mice upon high-fat feeding. Mechanistically, RIP140 was found to require the nuclear receptor ERRalpha to regulate hexose uptake and mitochondrial proteins SDHB and CoxVb, although it likely acts through other nuclear receptors as well. We conclude that RIP140 is a major suppressor of adipocyte oxidative metabolism and mitochondrial biogenesis, as well as a negative regulator of whole-body glucose tolerance and energy expenditure in mice. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16374519&query_hl=1 ER - TY - JFULL T1 - Hypoxia amplifies the proliferative capacity of distal human pulmonary artery smooth-muscle cells. A1 - Growcott, EJ A1 - Banner, KH A1 - Wharton, J J1 - Chest Y1 - 2005/12// VL - 128 SN - 0012-3692 SP - 600S EP - 601S L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16373856&query_hl=1 ER - TY - JFULL T1 - Defective modification of mannose residues by terminal phosphoethanolamine underlies inherited GPI deficiency. A1 - Almeida, AM A1 - Hillmen, P A1 - Richards, SJ A1 - Crawford, DH A1 - Baker, A A1 - Ferguson, MA A1 - Roberts, IA A1 - Layton, DM A1 - Karadimitris, A J1 - BLOOD Y1 - 2005/11/16/ VL - 106 SN - 0006-4971 SP - 41A EP - 41A ER - TY - JFULL T1 - Human CD34(+) haematopoietic stem cells (HSC) from umbilical cord blood display an endodermal phenotype when exposed to Activin A in vitro. A1 - Albera, C A1 - Bishop, AE A1 - Polak, JM A1 - Panoskaltsis, N J1 - BLOOD Y1 - 2005/11/16/ VL - 106 SN - 0006-4971 SP - 484A EP - 484A ER - TY - JFULL T1 - SALL4, a novel oncogene induces myelodysplastic syndrome and acute myeloid leukemia via Wnt/beta-catenin pathway. A1 - Chai, L A1 - Cui, W A1 - Chang, JC A1 - Di, CH A1 - Amin, H A1 - Ma, YP J1 - BLOOD Y1 - 2005/11/16/ VL - 106 SN - 0006-4971 SP - 397A EP - 397A ER - TY - JFULL T1 - Regulation of hematopoiesis in vitro and in vivo by invariant NKT cells. A1 - Kotsianidis, I A1 - Silk, JD A1 - Patterson, S A1 - Almeida, A A1 - Tsatalas, C A1 - Bourikas, G A1 - Cerundolo, V A1 - Roberts, IAG A1 - Karadimitris, A J1 - BLOOD Y1 - 2005/11/16/ VL - 106 SN - 0006-4971 SP - 641A EP - 642A ER - TY - JFULL T1 - High pH lysis allows immunoprecipitation of p210(BCR-ABL) in primary chronic myeloid leukaemia. A1 - Patel, H A1 - Marley, SB A1 - Gordon, MY J1 - BLOOD Y1 - 2005/11/16/ VL - 106 SN - 0006-4971 SP - 297B EP - 297B ER - TY - JFULL T1 - RIP140-targeted repression of gene expression in adipocytes. A1 - Christian, M A1 - Kiskinis, E A1 - Debevec, D A1 - Leonardsson, G A1 - White, R A1 - Parker, MG J1 - Mol Cell Biol Y1 - 2005/11// VL - 25 SN - 0270-7306 SP - 9383 EP - 9391 N2 - Ligand-dependent repression of nuclear receptor activity forms a novel mechanism for regulating gene expression. To investigate the intrinsic role of the corepressor RIP140, we have monitored gene expression profiles in cells that express or lack the RIP140 gene and that can be induced to undergo adipogenesis in vitro. In contrast to normal white adipose tissue and in vitro-differentiated wild-type adipocytes, RIP140-null cells show elevated energy expenditure and express high levels of the uncoupling protein 1 gene (Ucp1), carnitine palmitoyltransferase 1b, and the cell-death-inducing DFF45-like effector A. Conversely, all these changes are abrogated by the reexpression of RIP140. Analysis of the Ucp1 promoter showed RIP140 recruitment to a key enhancer element, demonstrating a direct role in repressing gene expression. Therefore, reduction in the levels of RIP140 or prevention of its recruitment to nuclear receptors may provide novel mechanisms for the control of energy expenditure in adipose cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16227589&query_hl=1 ER - TY - JFULL T1 - Adult bone marrow-derived stem cells and the injured heart: just the beginning? A1 - Dimarakis, I A1 - Habib, NA A1 - Gordon, MY J1 - Eur J Cardiothorac Surg Y1 - 2005/11// VL - 28 SN - 1010-7940 SP - 665 EP - 676 N2 - Coronary artery disease leading to ischaemia of the human myocardium is one of the chief causes of morbidity and mortality in the western world. Cellular transplantation has recently been proposed as a novel alternative treatment modality. Adult bone marrow-derived autologous cells are one of the key cell types under investigation in both the experimental and clinical setting. A range of theories has been proposed with regard to the observed myocardial function improvement in both human and animal studies. A concerted interest in scientific questions needs to be constructed on the regenerative information made available throughout the last years. It is only now that we begin to fully understand this fast growing body of research and how it may have wide ranging implications for the treatment of ischemic heart disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16194607&query_hl=1 ER - TY - JFULL T1 - The effects of great cardiac vein occlusion on cardiac pacing threshold and connexin 43 expression in myocardium in a pig model A1 - Cui, W A1 - Liu, F A1 - Hao, Y A1 - Xie, R A1 - Du, J J1 - CIRCULATION Y1 - 2005/10/25/ VL - 112 SN - 0009-7322 SP - U785 EP - U785 ER - TY - JFULL T1 - Direct intramyocardial injection, but not intracoronary infusion of bone marrow mononuclear cells induces ventricular arrhythmias in chronic ischemic heart failure A1 - Fukushima, S A1 - Brindley, G A1 - Yamahara, K A1 - Coppen, SR A1 - Lee, J A1 - Varela-Carver, A A1 - Yacoub, MH A1 - Terracciano, CM A1 - Suzuki, K J1 - CIRCULATION Y1 - 2005/10/25/ VL - 112 SN - 0009-7322 SP - U695 EP - U695 ER - TY - JFULL T1 - Abnormal erythroid cell development and erythropoietin hypo-responsiveness in chronic heart failure: A novel analysis of underlying cellular and molecular mechanisms A1 - Okonko, DO A1 - Marley, SB A1 - Crosato, M A1 - Anker, SD A1 - Gordon, MY A1 - Poole-Wilson, PA J1 - CIRCULATION Y1 - 2005/10/25/ VL - 112 SN - 0009-7322 SP - U822 EP - U822 ER - TY - JFULL T1 - Human invariant NKT cells are required for effective in vitro alloresponses. A1 - Patterson, S A1 - Kotsianidis, I A1 - Almeida, A A1 - Politou, M A1 - Rahemtulla, A A1 - Matthew, B A1 - Schmidt, RR A1 - Cerundolo, V A1 - Roberts, IA A1 - Karadimitris, A J1 - J Immunol Y1 - 2005/10/15/ VL - 175 SN - 0022-1767 SP - 5087 EP - 5094 N2 - NKT cells are a small subset of regulatory T cells conserved in humans and mice. In humans they express the Valpha24Jalpha18 invariant chain (hence invariant NKT (iNKT) cells) and are restricted by the glycolipid-presenting molecule CD1d. In mice, iNKT cells may enhance or inhibit anti-infectious and antitumor T cell responses but suppress autoimmune and alloreactive responses. We postulated that iNKT cells might also modulate human alloreactive responses. Using MLR assays we demonstrate that in the presence of the CD1d-presented glycolipid alpha-galactosylceramide (alphaGC) alloreactivity is enhanced (37 +/- 12%; p < 0.001) in an iNKT cell-dependent manner. iNKT cells are activated early during the course of the MLR, presumably by natural ligands. In MLR performed without exogenous ligands, depletion of iNKT cells significantly diminished the alloresponse in terms of proliferation (58.8 +/- 24%; p < 0.001) and IFN-gamma secretion (43.2 +/- 15.2%; p < 0.001). Importantly, adding back fresh iNKT cells restored the reactivity of iNKT cell-depleted MLR to near baseline levels. CD1d-blocking mAbs equally reduced the reactivity of the iNKT cell-replete and -depleted MLR compared with IgG control, indicating that the effect of iNKT cells in the in vitro alloresponse is CD1d-dependent. These findings suggest that human iNKT cells, although not essential for its development, can enhance the alloreactive response. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16210612&query_hl=1 ER - TY - JFULL T1 - Cholangiocarcinoma. A1 - Khan, SA A1 - Thomas, HC A1 - Davidson, BR A1 - Taylor-Robinson, SD J1 - Lancet Y1 - 2005/10/08/ VL - 366 SN - 1474-547X SP - 1303 EP - 1314 N2 - Cholangiocarcinoma is a devastating malignancy that presents late, is notoriously difficult to diagnose, and is associated with a high mortality. The incidence of intrahepatic cholangiocarcinoma is increasing worldwide. The cause for this rise is unclear, although it could be related to an interplay between predisposing genetic factors and environmental triggers. MRI and CT with endoscopic ultrasound and PET provide useful diagnostic information in certain patients. Surgical resection is the only chance for cure, with results depending on careful technique and patient selection. Data suggest that liver transplantation could offer long-term survival in selected patients when combined with neoadjuvant chemoradiotherapy. Chemotherapy and radiotherapy have been ineffective for patients with inoperable tumours. For most of these patients biliary drainage is the mainstay of palliation. However, controversy exists over the type and positioning of biliary stents. Photodynamic treatment is a new palliative technique that might improve quality of life. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16214602&query_hl=1 ER - TY - JFULL T1 - Differentiation of human embryonic stem cells to neural lineages in adherent culture by blocking bone morphogenetic protein signaling. A1 - Gerrard, L A1 - Rodgers, L A1 - Cui, W J1 - Stem Cells Y1 - 2005/10// VL - 23 SN - 1066-5099 SP - 1234 EP - 1241 N2 - Human embryonic stem cells (hESCs) have extensive self-renewal capacity and are competent to differentiate into any cell type of the body. They are valuable not only for the study of early human development but also for regenerative medicine. However, how to direct differentiation of hESCs along a particular lineage pathway to a specific cell type remains a challenge. Although hESCs have been shown to differentiate in vitro into neural progenitors, the factors controlling their differentiation are poorly understood. In this study, we report the development of an in vitro adherent culture system to efficiently generate neural progenitors in which neither multicellular aggregates nor stromal cells are required. We show that inhibition of bone morphogenetic protein signaling by its antagonist noggin is sufficient to block extraembryonic cell fate, transiently sustain Oct4 gene expression, and result in robust production of neural progenitors. Our findings will provide a platform for studying the molecular mechanism controlling neural differentiation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16002783&query_hl=1 ER - TY - JFULL T1 - Vivo and in vitro effect and mechanisms of cyclooxygenase-2 (COX-2)-independent growth inhibition of human hepatocellular carcinoma (HCC) cells by celecoxib A1 - Cui, W A1 - Yu, CH A1 - Tang, ZY A1 - Hu, KQ J1 - HEPATOLOGY Y1 - 2005/10// VL - 42 SN - 0270-9139 SP - 302A EP - 302A ER - TY - JFULL T1 - Inhibitory effects and mechanisms of silibinin on growth of human hepatoma cell lines A1 - Lah, J A1 - Cui, W A1 - Hu, KQ J1 - HEPATOLOGY Y1 - 2005/10// VL - 42 SN - 0270-9139 SP - 309A EP - 309A ER - TY - JFULL T1 - Distinctive nuclear organisation of centromeres and regions involved in pluripotency in human embryonic stem cells. A1 - Wiblin, AE A1 - Cui, W A1 - Clark, AJ A1 - Bickmore, WA J1 - J Cell Sci Y1 - 2005/09/01/ VL - 118 SN - 0021-9533 SP - 3861 EP - 3868 N2 - Nuclear organisation is thought to be important in regulating gene expression. Here we investigate whether human embryonic stem cells (hES) have a particular nuclear organisation, which could be important for maintaining their pluripotent state. We found that whereas the nuclei of hES cells have a general gene-density-related radial organisation of chromosomes, as is seen in differentiated cells, there are also distinctive localisations for chromosome regions and gene loci with a role in pluripotency. Chromosome 12p, a region of the human genome that contains clustered pluripotency genes including NANOG, has a more central nuclear localisation in ES cells than in differentiated cells. On chromosome 6p we find no overall change in nuclear chromosome position, but instead we detect a relocalisation of the OCT4 locus, to a position outside its chromosome territory. There is also a smaller proportion of centromeres located close to the nuclear periphery in hES cells compared to differentiated cells. We conclude that hES cell nuclei have a distinct nuclear architecture, especially at loci involved in maintaining pluripotency. Understanding this level of hES cell biology provides a framework within which other large-scale chromatin changes that may accompany differentiation can be considered. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16105879&query_hl=1 ER - TY - JFULL T1 - Multiple signaling defects in the absence of RIP140 impair both cumulus expansion and follicle rupture. A1 - Tullet, JM A1 - Pocock, V A1 - Steel, JH A1 - White, R A1 - Milligan, S A1 - Parker, MG J1 - Endocrinology Y1 - 2005/09// VL - 146 SN - 0013-7227 SP - 4127 EP - 4137 N2 - The nuclear receptor corepressor RIP140 is essential in the ovary for ovulation, but is not required for follicle growth and luteinization. To identify genes that may be subject to regulation by RIP140 or play a role in ovulation, we compared ovarian gene expression profiles in untreated immature wild-type and RIP140 null mice and after treatment with pregnant mare serum gonadotropin and human chorionic gonadotropin. Many genes involved in signaling, extracellular matrix formation, cell-cell attachment, and adhesion were aberrantly regulated in the absence of RIP140, varying according to the hormone status of the mice. Notable among these was the reduced expression of a number of genes that encode components of signaling pathways and matrix proteins required for cumulus expansion, a key remodeling process necessary for ovulation. Histological analysis confirmed that cumulus expansion in RIP140 null mice is reduced, oocyte detachment from the mural cell wall is impaired, and follicles fail to rupture in response to LH. Although the expression of many genes involved in cumulus cell expansion was reduced, there was a subset of genes involved in extracellular matrix formation and cell-cell interactions that was up-regulated and may interfere with ovarian tissue remodeling. We propose that widespread gene dysregulation in ovarian tissues in the absence of RIP140 leads to the anovulatory phenotype. This helps to define an important role for RIP140 in the regulation of multiple processes leading to ovulation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15919748&query_hl=1 ER - TY - JFULL T1 - Structure: Function characterization of ASP : C5L2 receptor interaction and relevance to obesity A1 - Cui, W A1 - Cianflone, K J1 - OBES RES Y1 - 2005/09// VL - 13 SN - 1071-7323 SP - A41 EP - A41 ER - TY - JFULL T1 - Modelling GPCR effects in the cardiomyocyte: A bridge from reconstituted systems to human heart A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2005/09// VL - 39 SN - 0022-2828 SP - 411 EP - 413 ER - TY - JFULL T1 - Stem cells. A1 - Vats, A A1 - Bielby, RC A1 - Tolley, NS A1 - Nerem, R A1 - Polak, JM J1 - Lancet Y1 - 2005/08/13/ VL - 366 SN - 1474-547X SP - 592 EP - 602 N2 - Stem cells derived from adult and embryonic sources have great therapeutic potential, but much research is still needed before their clinical use becomes commonplace. There is debate about whether adult stem cells can be used instead of those derived from embryos. Rationalisation is needed but can be exercised only once the various cells have been carefully compared and contrasted under appropriate experimental conditions. Some characteristics that might help resolve the issue of cell source can already be applied to the debate. Accessibility is important; some adult cells, such as neural stem cells, are difficult to obtain, at least from living donors. Other factors include the frequency and abundance of adult stem cells and their numbers and potency, which might decline with age or be affected by disease. For embryonic stem cells, ethical concerns have been raised, and the proposed practice of therapeutic cloning tends to be misrepresented in the lay media. For both adult and embryonic stem cells, stability, potential to transmit harmful pathogens or genetic mutations, and risk of forming unwanted tissues or even teratocarcinomas have yet to be fully assessed. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16099296&query_hl=1 ER - TY - JFULL T1 - Use of green fluorescent fusion protein to track activation of the transcription factor osterix during early osteoblast differentiation. A1 - Tai, G A1 - Christodoulou, I A1 - Bishop, AE A1 - Polak, JM J1 - Biochem Biophys Res Commun Y1 - 2005/08/12/ VL - 333 SN - 0006-291X SP - 1116 EP - 1122 N2 - Osterix (Osx) is a transcription factor required for the differentiation of preosteoblasts into fully functioning osteoblasts. However, the pattern of Osx activation during preosteoblast differentiation and maturation has not been clearly defined. Our aim was to study Osx activation during these processes in osteoblasts differentiating from murine and human embryonic stem cells (ESC). To do this, we constructed an Osx-GFP fusion protein reporter system to track Osx translocation within the cells. The distribution of Osx-GFP at representative stages of differentiation was also investigated by screening primary osteoblasts, mesenchymal stem cells, synoviocytes, and pre-adipocytes. Our experiments revealed that Osx-GFP protein was detectable in the cytoplasm of cultured, differentiated ESC 4 days after plating of enzymatically dispersed embryoid bodies. Osterix-GFP protein became translocated into the nucleus on day 7 following transfer of differentiated ESC to osteogenic medium. After 14 days of differentiation, cells showing nuclear translocation of Osx-GFP formed rudimentary bone nodules that continued to increase in number over the following weeks (through day 21). We also found that Osx translocated into the nuclei of mesenchymal stem cells (C3H10T1/2) and pre-osteoblasts (MC3T3-E1) and showed partial activation in pre-adipocytes (MC3T3-L1). These data suggest that Osx activation occurs at a very early point in the differentiation of the mesenchymal-osteoblastic lineage. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15979565&query_hl=1 ER - TY - JFULL T1 - p53 Mutations in human cholangiocarcinoma: a review. A1 - Khan, SA A1 - Thomas, HC A1 - Toledano, MB A1 - Cox, IJ A1 - Taylor-Robinson, SD J1 - Liver Int Y1 - 2005/08// VL - 25 SN - 1478-3223 SP - 704 EP - 716 N2 - The reported mortality from intrahepatic bile duct tumours is increasing markedly in industrialised countries, for reasons that remain unknown. Inactivation of the tumour suppressor gene p53, is the commonest genetic abnormality in human cancer and has been implicated in the genesis of cholangiocarcinoma in various immunohistochemical and molecular epidemiological investigations, including gene sequencing studies. The structure and function of p53 and its role in linking cancer to specific carcinogens by way of mutational signatures is reviewed. The findings of previous p53 studies and their relevance in human cholangiocarcinoma are summarised. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15998419&query_hl=1 ER - TY - JFULL T1 - Phosphodiesterase type 5 and high altitude pulmonary hypertension. A1 - Aldashev, AA A1 - Kojonazarov, BK A1 - Amatov, TA A1 - Sooronbaev, TM A1 - Mirrakhimov, MM A1 - Morrell, NW A1 - Wharton, J A1 - Wilkins, MR J1 - Thorax Y1 - 2005/08// VL - 60 SN - 0040-6376 SP - 683 EP - 687 N2 - BACKGROUND: This study explored phosphodiesterase type 5 (PDE5) inhibition as a strategy for treating high altitude pulmonary arterial hypertension (HAPH). METHODS: 689 subjects (313 men) of mean (SD) age 44 (0.6) years living above 2500 m were screened for HAPH by medical examination and electrocardiography, and 188 (27%) met the criteria for right ventricular hypertrophy. 44 underwent cardiac catheterisation and 29 (66%) had a resting mean pulmonary artery pressure (PAP) above 25 mmHg. 22 patients with a raised mean PAP were randomised to receive sildenafil (25 or 100 mg) or matching placebo taken 8 hourly for 12 weeks. RESULTS: At 3 months, patients on sildenafil 25 mg 8 hourly (n = 9) had a significantly (p = 0.018) lower mean PAP (-6.9 mmHg) at the end of the dosing interval than those on placebo (n = 8) (95% CI -12.4 to -1.3). The treatment effect for sildenafil 100 mg 8 hourly (n = 5) compared with placebo was -6.4 mm Hg (95% CI -12.9 to 0.1). Both doses improved 6 minute walk distance, the lower dose by 45.4 m (95% CI 11.5 to 79.4; p = 0.011) and the higher dose by 40.0 m (95% CI 0.2 to 79.8; p = 0.049). Sildenafil was well tolerated. Necroscopic lung specimens from three subjects with HAPH showed abundant PDE5 in the muscular coat of remodelled pulmonary arterioles. CONCLUSIONS: PDE5 is an attractive drug target for the treatment of HAPH and a larger study of the long term effects of PDE5 inhibition in HAPH is warranted. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16061711&query_hl=1 ER - TY - JFULL T1 - Hypothermia and amiloride preserve energetics in a neonatal brain slice model. A1 - Robertson, NJ A1 - Bhakoo, K A1 - Puri, BK A1 - Edwards, AD A1 - Cox, IJ J1 - Pediatr Res Y1 - 2005/08// VL - 58 SN - 0031-3998 SP - 288 EP - 296 N2 - A period of secondary energy failure consisting of a decline in phosphocreatine/inorganic phosphate (PCr/Pi), a rise in brain lactate, and alkaline intracellular pH (pH(i)) has been described in infants with neonatal encephalopathy. Strategies that ameliorate this energy failure may be neuroprotective. We hypothesized that a neonatal rat brain slice model undergoes a progressive decline in energetics, which can be ameliorated with hypothermia or amiloride. Interleaved phosphorus ((31)P) and proton ((1)H) magnetic resonance (MR) spectra were obtained from 350 microm neonatal rat brain slices over 8 h in a bicarbonate buffer at 37 degrees C and at 32 degrees C in 7- and 14-d models. (31)P MR spectra were obtained with amiloride in a bicarbonate-free buffer at 37 degrees C in the 14-d model. Findings were similar in 7- and 14-d models. In the 14-d model, there was a Pi doublet structure corresponding to alkaline pH(i) values of 7.50 +/- 0.02 and 7.21 +/- 0.04. Compared with the stabilized baseline of 100, at 5 h PCr/Pi was 65 +/- 6.3 and lactate/NAA was 187 +/- 3 at 37 degrees C, but PCr/Pi and lactate/NAA were not significantly different from baseline at 32 degrees C. Nucleotide triphosphate (NTP)/phosphomonoester (PME) was 0.93 +/- 0.23 at 37 degrees C and 1.81 +/- 0.21 at 32 degrees C at 5 h. With amiloride exposure in the 14-d model, baseline pH(i) values were 7.25 +/- 0.09 and 6.98 +/- 0.02 and NTP/PME was 1.81 +/- 0.05; these parameters were not significantly different at 5 h. Our interpretation of these findings is that the brain slice model underwent secondary energy failure, which was delayed with hypothermia or amiloride. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16006423&query_hl=1 ER - TY - JFULL T1 - Embryonic stem cells and tissue engineering: delivering stem cells to the clinic. A1 - Vats, A A1 - Tolley, NS A1 - Bishop, AE A1 - Polak, JM J1 - J R Soc Med Y1 - 2005/08// VL - 98 SN - 0141-0768 SP - 346 EP - 350 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16055897&query_hl=1 ER - TY - JFULL T1 - Coculture of embryonic stem cells with pulmonary mesenchyme: a microenvironment that promotes differentiation of pulmonary epithelium. A1 - Van Vranken, BE A1 - Romanska, HM A1 - Polak, JM A1 - Rippon, HJ A1 - Shannon, JM A1 - Bishop, AE J1 - Tissue Eng Y1 - 2005/07// VL - 11 SN - 1076-3279 SP - 1177 EP - 1187 N2 - Coculture of stem/progenitor cells with mature cells or tissues can drive their differentiation toward required lineages. Thus, we hypothesized that coculture of murine embryonic stem (ES) cells with embryonic mesenchyme from distal lung promotes the differentiation of pneumocytes. Murine ES cells were differentiated to embryoid bodies (EBs) and cultured for 5 or 12 days with pulmonary mesenchyme from embryonic day 11.5 or 13.5 murine embryos, in direct contact or separated by a membrane. Controls included EBs cultured alone or with embryonic gut mesenchyme. Histology revealed epithelium-lined channels in directly cocultured EBs, whereas EBs grown alone showed little structural organization. The lining cells expressed cytokeratin and thyroid transcription factor 1, an early developmental marker in pulmonary epithelium. Differentiation of type II pneumocytes specifically was demonstrated by the presence of surfactant protein C (SP-C) in some of the epithelial cells. None of these markers was seen in EBs cultured alone or with embryonic gut mesenchyme. Indirect coculture of EBs with lung mesenchyme resulted in a 14-fold increase in SP-C gene expression. Thus, provision of an appropriate microenvironment, in the form of pulmonary mesenchyme, appears to promote the differentiation of ES cells toward lung epithelium. Our findings may have applications in regenerative medicine strategies and the engineering of lung tissue. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16144454&query_hl=1 ER - TY - JFULL T1 - Chronic myeloid leukaemia: stem cell derived but progenitor cell driven. A1 - Marley, SB A1 - Gordon, MY J1 - Clin Sci (Lond) Y1 - 2005/07// VL - 109 SN - 0143-5221 SP - 13 EP - 25 N2 - The biology of CML (chronic myeloid leukaemia) has been extensively investigated as the disease is a paradigm of neoplasms induced when a translocation results in expression of a novel fusion protein, in this instance p210(BCR-ABL). Although CML manifests itself principally as unregulated expansion of the myeloid lineage, the lesion is present in the stem cell population and it has long been assumed that disregulated stem cell kinetics must underlie the basic pathology of the disease. In this review, we present evidence that, in normal haemopoiesis, less primitive precursor cells retain considerable flexibility in their capacity to undergo self-renewal, allowing them to maintain lineage-specific homoeostasis without inflicting proliferative stress upon the stem cell population. This mechanism is dysregulated in CML and we have developed a self-renewal assay for CFU-GM (colony-forming unit-granulocyte/macrophage) which demonstrates that, in CML, the PI (proliferative index) of the myeloid progenitor cell population is increased. The ability to measure the PI as an endpoint of p210(BCR-ABL) expression gives considerable versatility to the in vitro investigation of putative therapeutic regimes in CML. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15966868&query_hl=1 ER - TY - JFULL T1 - Repopulation of human pulmonary epithelium by bone marrow cells: a potential means to promote repair. A1 - Albera, C A1 - Polak, JM A1 - Janes, S A1 - Griffiths, MJ A1 - Alison, MR A1 - Wright, NA A1 - Navaratnarasah, S A1 - Poulsom, R A1 - Jeffery, R A1 - Fisher, C A1 - Burke, M A1 - Bishop, AE J1 - Tissue Eng Y1 - 2005/07// VL - 11 SN - 1076-3279 SP - 1115 EP - 1121 N2 - After lung injury and damage to the alveolar epithelium, the underlying basement membranes become exposed. Proliferation of type II pneumocytes and their differentiation to the type I phenotype have been considered to be the mechanism by which repopulation of the alveolar epithelium occurs. A growing body of evidence has shown that tissues can be repaired by cells acquired via the circulation. For the lung, bone marrow stem cells have been shown in mice to regenerate epithelium as well as give rise to the expected mesodermal derivatives. We hypothesized that extrapulmonary cells, including those from the bone marrow, can contribute to the reepithelialization of human alveoli. To investigate this, we examined samples of peripheral lung from patients who had undergone cross-gender transplantation of lung or bone marrow. Thus, archival blocks of peripheral lung were analyzed from male patients (surgical samples, n = 8) who had received a lung transplant from a female donor and female patients (postmortem samples, n = 3) who had male bone marrow transplants. In both cases, male cells were identified in the female lungs by Y chromosome in situ hybridization. Male cells could be identified in the alveolar epithelium where, in the better preserved, transplanted lungs, it was possible to show that some had differentiated to type II pneumocytes. In addition, Y chromosomes were found to be widespread in cells of mesenchymal lineage, including macrophages and endothelial cells. Concomitant visualization of Y and X chromosomes, using fluorescence immunolabeling, yielded no evidence of cellular fusion, although the poor quality of the autopsy samples studied meant that the possibility could not be excluded. These observations suggest that, as occurs in rodents, the epithelium of the adult human lung has the capacity to renew itself, using cells recruited from extrapulmonary sources, including the bone marrow. This finding could provide new therapeutic opportunities for a range of pulmonary diseases by providing means to repair the lung and a novel route for gene therapy. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16144447&query_hl=1 ER - TY - JFULL T1 - Dose- and time-dependent effect of bioactive gel-glass ionic-dissolution products on human fetal osteoblast-specific gene expression. A1 - Christodoulou, I A1 - Buttery, LD A1 - Saravanapavan, P A1 - Tai, G A1 - Hench, LL A1 - Polak, JM J1 - J Biomed Mater Res B Appl Biomater Y1 - 2005/07// VL - 74 SN - 1552-4973 SP - 529 EP - 537 N2 - Bioactive glasses dissolve upon immersion in culture medium, and release their constitutive ions into solution. There has been some evidence suggesting that these ionic-dissolution products influence osteoblast-specific processes. Here, the effect of 58S sol-gel-derived bioactive glass (60% SiO(2), 36% CaO, 4% P(2)O(5), in molar percentage) on primary osteoblasts derived from human fetal long bone explant cultures is investigated, and it is hypothesized that critical concentrations of sol-gel-dissolution products (consisting of a combination of simple inorganic ions) can enhance osteoblast phenotype in vitro by affecting the expression of a number of genes associated with the differentiation and extracellular matrix deposition processes. Cells were exposed to a range of 58S dosages continuously for a period of 4-14 days in monolayer cultures. Quantitative real-time RT-PCR analysis of a panel of osteoblast-specific markers showed a varied gene expression pattern in response to the material. The highest concentration of Ca and Si tested (96 and 50 ppm, respectively) promoted upregulation of gene expression for most markers (including alkaline phosphatase, osteocalcin, and osteopontin) at the latest time point, compared to non-58S-treated control, although this observation was not statistically significant. The same 58S concentration produced higher ALP activity levels and increased proliferation throughout the culture period, compared to lower dosages tested; however, the results generated were again not statistically significant. The data overall suggest that no significant effect can be ascribed to the ionic products of 58S bioactive gel-glass dissolution tested here and their ability to stimulate osteoblastic marker gene expression. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15889438&query_hl=1 ER - TY - JFULL T1 - Proton and phosphorus-31 nuclear magnetic resonance spectroscopy of human bile in hepatopancreaticobiliary cancer. A1 - Khan, SA A1 - Cox, IJ A1 - Thillainayagam, AV A1 - Bansi, DS A1 - Thomas, HC A1 - Taylor-Robinson, SD J1 - Eur J Gastroenterol Hepatol Y1 - 2005/07// VL - 17 SN - 0954-691X SP - 733 EP - 738 N2 - OBJECTIVE: Hepatopancreaticobiliary cancers can be difficult to diagnose. Nuclear magnetic resonance (NMR) spectroscopy provides non-invasive information on phospholipid metabolism, and previous studies of liver tissue have highlighted changes in phospholipids in malignancy. We hypothesised that in-vitro NMR spectroscopy of human bile may provide independent diagnostic indices in cancer management through an assessment of the phospholipid content. DESIGN AND METHODS: Bile samples from 24 patients were collected at endoscopic retrograde cholangiopancreatography and from one subject at cholecystectomy. Thirteen patients had cancer: pancreatic carcinoma (eight), cholangiocarcinoma (three) and metastatic liver disease (two). The remaining 12 patients had non-malignant pathology. In-vitro proton (H) and phosphorus-31 (P) NMR spectra were obtained from all samples using an 11.7 Tesla NMR spectroscopy system. RESULTS: Complementary information was obtained from the H and P NMR spectra. Signals were assigned to phosphatidylcholine in both H and P NMR spectra. Phosphatidylcholine levels were significantly reduced in the bile from cancer patients when compared with bile from non-cancer patients (P=0.007). CONCLUSION: These preliminary studies suggest that H and P NMR spectroscopy of bile may be used to detect differences in phospholipid content between cancer and non-cancer patients. This may have implications for the development of novel diagnostic strategies in hepatopancreaticobiliary cancers. Further larger-scale studies are warranted. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15947550&query_hl=1 ER - TY - JFULL T1 - Lipid rafts play a role in the formation of an adhesive junction between homotypically aggregated CD34-positive cells A1 - Bullock, TE A1 - Marley, SB A1 - Gordon, MY J1 - EXP HEMATOL Y1 - 2005/07// VL - 33 SN - 0301-472X SP - 94 EP - 94 ER - TY - JFULL T1 - Antiproliferative effects of phosphodiesterase type 5 inhibition in human pulmonary artery cells. A1 - Wharton, J A1 - Strange, JW A1 - Møller, GM A1 - Growcott, EJ A1 - Ren, X A1 - Franklyn, AP A1 - Phillips, SC A1 - Wilkins, MR J1 - Am J Respir Crit Care Med Y1 - 2005/07/01/ VL - 172 SN - 1073-449X SP - 105 EP - 113 N2 - RATIONALE: Phosphodiesterase Type 5 (PDE5) inhibition represents a novel strategy for the treatment of pulmonary hypertension. OBJECTIVES: Our aim was to establish the distribution of PDE5 in the pulmonary vasculature and effects of PDE5 inhibition on pulmonary artery smooth muscle cells (PASMCs). METHODS AND MEASUREMENTS: PDE5 expression was examined by immunohistochemistry and Western blotting, in both normal and hypertensive lung tissues. DNA synthesis, proliferation, PDE activity, and apoptosis were measured in distal human PASMCs treated with soluble guanylyl cyclase activators (nitric oxide donors and BAY41-2272) and sildenafil. MAIN RESULTS: Cells containing PDE5 and alpha-smooth muscle actin occurred throughout the pulmonary vasculature, including obstructive intimal lesions. Three molecular forms of PDE5 were identified and protein expression was greater in hypertensive than control lung tissue. Most cyclic guanosine monophosphate hydrolysis (about 80%) in cultured cells was attributed to PDE5. Sildenafil induced a greater elevation of intracellular cyclic guanosine monophosphate levels compared with nitric oxide donors and BAY41-2272 (about 10-fold versus about 2-fold) and cotreatment had a synergistic effect, increasing cyclic nucleotide levels up to 50-fold. Dual stimulation of soluble guanylyl cyclase and inhibition of PDE5 activities also had significant downstream effects, increasing phosphorylation of vasodilator-stimulated phosphoprotein, reducing DNA synthesis and cell proliferation, and stimulating apoptosis, and these effects were mimicked by cyclic guanosine monophosphate analogs. CONCLUSIONS: Phosphodiesterase Type 5 is the main factor regulating cyclic guanosine monophosphate hydrolysis and downstream signaling in human PASMCs. The antiproliferative effects of this signaling pathway may be significant in the chronic treatment of pulmonary hypertension with PDE5 inhibitors such as sildenafil. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15817798&query_hl=1 ER - TY - JFULL T1 - A subpopulation of CD34+stem cells with potential to repopulate tissue A1 - Gordon, MY A1 - Levicar, N A1 - Al-Allaf, F A1 - Dimarakis, I A1 - Thalji, T A1 - Welsh, J A1 - Alexandre, E A1 - Habib, NA J1 - EXP HEMATOL Y1 - 2005/07// VL - 33 SN - 0301-472X SP - 109 EP - 110 ER - TY - JFULL T1 - The cytokine environment influences clonal dominance and response to imatinib by CML 254 progenitor cells in vitro A1 - Ojo, M A1 - Gordon, MY A1 - Marley, SB J1 - EXP HEMATOL Y1 - 2005/07// VL - 33 SN - 0301-472X SP - 104 EP - 104 ER - TY - JFULL T1 - The failing cardiomyocyte. A1 - Harding, SE J1 - Heart Fail Clin Y1 - 2005/07// VL - 1 SN - 1551-7136 SP - 171 EP - 181 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17386844&query_hl=1 ER - TY - JFULL T1 - Targeting genes and cells in the progression to heart failure. A1 - Sato, M A1 - Fuller, SJ A1 - Hajjar, RJ A1 - Harding, SE J1 - Heart Fail Clin Y1 - 2005/07// VL - 1 SN - 1551-7136 SP - 287 EP - 301 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17386853&query_hl=1 ER - TY - JFULL T1 - Interaction between in vivo gene transfer of phospholamban antisense and catecholamine infusion in rat heart A1 - Sato, M A1 - O'Gara, P A1 - Fuller, SJ A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2005/06// VL - 38 SN - 0022-2828 SP - 1064 EP - 1064 ER - TY - JFULL T1 - Enhancement of dendritic cell production and function by Fms-like tyrosine kinase-3 ligand decreases susceptibility of mice to burn wound infection. A1 - Toliver-Kinsky, T A1 - Cui, W A1 - Murphey, E A1 - Sherwood, E J1 - SHOCK Y1 - 2005/06// VL - 23 SN - 1073-2322 SP - 77 EP - 77 ER - TY - JFULL T1 - Enhancement of adenoviral gene transfer to adult rat cardiomyocytes in vivo by immobilization and ultrasound treatment of the heart. A1 - Sato, M A1 - O'Gara, P A1 - Harding, SE A1 - Fuller, SJ J1 - Gene Ther Y1 - 2005/06// VL - 12 SN - 0969-7128 SP - 936 EP - 941 N2 - Direct injection of adenoviral vectors into ventricular myocardium in vivo produces local transfection of cells including cardiomyocytes. The use of vectors coexpressing GFP with the gene of interest allows subsequent identification of transfected myocytes isolated from the heart some days later, and examination of their function in cell bath experiments. We have injected vectors for antisense to phospholamban, or a control virus for expression of GFP only, into adult rat heart in vivo and then removed the heart and isolated ventricular myocytes 7 days later. Brief immobilization of the ventricle during and after injection using a haemoclip increased the number of transfected rod-shaped, viable myocytes from 1.7 +/- 0.8% (n = 8) to 5.6 +/- 0.8% (n = 9). This was further increased to 13.2 +/- 1.1% (n = 8) by the application of ultrasound pulses to the site before and after injection. Phospholamban antisense increased contraction amplitude and accelerated myocyte relengthening or decline of the Ca(2+) transient in transfected myocytes, while GFP control did not. Qualitative and quantitative effects of phospholamban downregulation were comparable between in vivo and in vitro transfections. This technique will have a number of uses, including production of transfected myocytes without the problem of culture-induced changes in contractility. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15759019&query_hl=1 ER - TY - JFULL T1 - Interaction of Na+/Ca2+ exchanger with B2Ar/Gi pathway in rat ventricular myocytes A1 - Zheng, Z A1 - O'Gara, P A1 - Lee, J A1 - Terracciano, CMN A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2005/06// VL - 38 SN - 0022-2828 SP - 1084 EP - 1085 ER - TY - JFULL T1 - Induction of distal lung epithelial markers by stepwise differentiation of mouse embryonic stem cells A1 - Rippon, HJ A1 - Polak, JM A1 - Bishop, AE J1 - INT J EXP PATHOL Y1 - 2005/06// VL - 86 SN - 0959-9673 SP - A21 EP - A21 ER - TY - JFULL T1 - Myocardial depression by IL-6 is reversed by P38 MAP kinase inhibition A1 - Franklin, JL A1 - Pathan, N A1 - Levin, M A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2005/06// VL - 38 SN - 0022-2828 SP - 1021 EP - 1021 ER - TY - JFULL T1 - Cell extract-derived differentiation of embryonic stem cells. A1 - Qin, M A1 - Tai, G A1 - Collas, P A1 - Polak, JM A1 - Bishop, AE J1 - Stem Cells Y1 - 2005/06// VL - 23 SN - 1066-5099 SP - 712 EP - 718 N2 - Various means have been used to encourage the differentiation of embryonic stem cells (ESCs) toward specific lineages, including growth factor administration, genetic modification, and coculture with relevant cells/tissues. Cell extract-based reprogramming has recently been used to derive mature cells from nonrelated phenotypes. In this communication, we tested whether this in vitro reprogramming approach can be used to direct ESC differentiation. Permeabilized murine ESCs exposed to extracts of murine type II pneumocytes showed increased expression of surfactant protein C and its corresponding mRNA, reflecting enhanced differentiation of pneumocytes. Subsequent differentiation to a type I phenotype was demonstrated by expression of aquaporin 5. Pneumocyte formation occurred quicker than with growth factor-induced differentiation. Our findings establish that ESCs can be differentiated in vitro using cellular extracts. This model provides a tool for analysis of the key factors involved in the differentiation of ESCs to type II pneumocytes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15917467&query_hl=1 ER - TY - JFULL T1 - Effects of endotoxic shock on neuronal NOS and calcium transients in rat cardiac myocytes. A1 - Comini, L A1 - Boraso, A A1 - Bachetti, T A1 - Bernocchi, P A1 - Pasini, E A1 - Bastianon, D A1 - Curello, S A1 - Terracciano, CM A1 - Ceconi, C A1 - Ferrari, R J1 - Pharmacol Res Y1 - 2005/05// VL - 51 SN - 1043-6618 SP - 409 EP - 417 N2 - OBJECTIVE: The effects of endotoxic shock on transcriptional and translational pattern of nitric oxide synthase isoforms (NOSs) and cytoplasmic calcium were investigated. METHODS: Male SD rats injected with lipopolysaccharides or saline were sacrificed after 6 and 20 h. Cardiac myocytes were enzimatically isolated from the excised hearts and evaluated for: (1) expression of constitutive (e and n) and inducible (i) NOSs by RT-PCR; (2) NOSs protein levels by Western blot, enzymatic activities by a radioimmunometric assay and nitric oxide metabolites by spectrophotometry; (3) calcium transients by Indo-1 fluorescence. RESULTS: Increase in iNOS mRNA, and decrease in e and nNOS mRNAs were observed in cardiac myocytes isolated 6h after LPS injection with recovery to basal levels at 20 h. Significant down-regulation of e and nNOS protein levels (p < 0.01) and calcium-dependent activity (p < 0.05) were detected at 20 h. Serum TNF-alpha increased after 6 and 20 h (p < 0.05), whereas NO metabolites rose only after 20 h (p < 0.0001). The diastolic calcium increased 6 h from LPS injection (p < 0.0001) and remained significantly higher after 20 h. Calcium transients amplitude was not affected by LPS injection. CONCLUSIONS: Endotoxic shock stimulates iNOS and down-regulates expression of nNOS in purified cardiac myocytes, but endogenous NO production does not likely affect calcium transients. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15749455&query_hl=1 ER - TY - JFULL T1 - Targeting primary human leukaemia cells with RNA interference: Bcr-Abl targeting inhibits myeloid progenitor self-renewal in chronic myeloid leukaemia cells A1 - Withey, JME A1 - Marley, SB A1 - Kaeda, J A1 - Harvey, AJ A1 - Crompton, MR A1 - Gordon, MY J1 - BRIT J HAEMATOL Y1 - 2005/05// VL - 129 SN - 0007-1048 SP - 377 EP - 380 N2 - We have investigated functional outcome of challenging primary chronic myeloid leukaemia (CML) cells with Bcr-Abl fusion sequence- directed RNA interference (RNAi). We targeted the Bcr-Abl b3a2 variant, by RNAi, in primary chronic phase CML cells, and detected strikingly reduced proliferation of myeloid precursor cells expressing this variant. Lack of an effect in cells expressing a distinct Bcr-Abl variant confirmed the specificity of the response. Through the functional targeting of an oncogene in primary human tumour cells, we have demonstrated that Bcr-Abl enhances CML progenitor cell amplification, and that RNAi may be suitable for development as a specific anti-leukaemia treatment. ER - TY - JFULL T1 - Influence of patency of the infarct-related artery before coronary intervention on myocardial salvage in patients with acute myocardial infarction. A1 - Cui, W A1 - Sarai, M A1 - Kondo, T A1 - Sato, T A1 - Oshima, K A1 - Naruse, H A1 - Ishii, J A1 - Mori, Y A1 - Hishida, H J1 - AM J CARDIOL Y1 - 2005/04/28/ VL - 95 SN - 0002-9149 SP - 6A EP - 6A ER - TY - JFULL T1 - D-dimer, but not the parameters of platelet activation, predicts myocardial injury after elective percutaneous coronary intervention. A1 - Cui, W A1 - Lu, JC A1 - Du, J A1 - Hao, YM A1 - Liu, F A1 - Li, YJ A1 - Xie, RQ J1 - AM J CARDIOL Y1 - 2005/04/28/ VL - 95 SN - 0002-9149 SP - 20A EP - 20A ER - TY - JFULL T1 - Effects of remote preconditioning induced by skeletal muscle ischemia on myocardial necrosis and apoptosis: Different roles of bradykinin and opioid receptors A1 - Cui, W A1 - Xie, RQ A1 - Hao, YM A1 - Du, GY A1 - Zhang, T A1 - Du, J A1 - Liu, F A1 - Li, BH A1 - Wu, JF J1 - AM J CARDIOL Y1 - 2005/04/28/ VL - 95 SN - 0002-9149 SP - 51A EP - 51A ER - TY - JFULL T1 - In vivo and in vitro nuclear magnetic resonance spectroscopy as a tool for investigating hepatobiliary disease: a review of H and P MRS applications. A1 - Khan, SA A1 - Cox, IJ A1 - Hamilton, G A1 - Thomas, HC A1 - Taylor-Robinson, SD J1 - Liver Int Y1 - 2005/04// VL - 25 SN - 1478-3223 SP - 273 EP - 281 N2 - Nuclear magnetic resonance (NMR) spectroscopy is a non-invasive technique, which allows the study of cellular biochemistry and metabolism. It is a diverse research tool, widely used by biochemists to investigate pathophysiological processes in vitro and, more recently, by physicians to determine disease abnormalities in vivo. This article reviews the basics of the NMR phenomenon and summarises previous research on the hepatobiliary system using both laboratory-based and clinical methodologies. The role of proton and phosphorus-31 ((31)P) NMR spectroscopy in the study of malignant and non-malignant liver disease and studies of bile composition are discussed. In vivo techniques (magnetic resonance spectroscopy, MRS) can be performed as an adjunct to standard MR examination of the liver. Although still primarily a research tool, the in vivo technique provides non-invasive biochemical information on disease severity and holds promise in its use to gauge response to treatment regimens. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15780050&query_hl=1 ER - TY - JFULL T1 - In vivo measurements of T1 relaxation times in mouse brain associated with different modes of systemic administration of manganese chloride. A1 - Kuo, YT A1 - Herlihy, AH A1 - So, PW A1 - Bhakoo, KK A1 - Bell, JD J1 - J Magn Reson Imaging Y1 - 2005/04// VL - 21 SN - 1053-1807 SP - 334 EP - 339 N2 - PURPOSE: To measure regional T1 and T2 values for normal C57Bl/6 mouse brain and changes in T1 after systemic administration of manganese chloride (MnCl2) at 9.4 T. MATERIALS AND METHODS: C57Bl/6 mice were anesthetized and baseline T1 and T2 measurements obtained prior to measurement of T1 after administration of MnCl2 at 9.4 T. MnCl2 was administered systemically either by the intravenous (IV), intraperitoneal (IP), or subcutaneous (SC) routes. T1 and T2 maps for each MRI transverse slice were generated using commercial software, and T1 and T2 values of white matter (WM), gray matter (GM), pituitary gland, and lateral ventricle were obtained. RESULTS: When compared with baseline values at low-field, significant lengthening of the T1 values was shown at 9.4 T, while no significant change was seen for T2 values. Significant T1 shortening of the normal mouse brain was observed following IV, IP, and SC administration of MnCl2, with IV and IP showing similar acute effects. Significant decreases in T1 values were seen for the pituitary gland and the ventricles 15 minutes after either IV or IP injection. GM showed greater uptake of the contrast agent than WM at 15 and 45 minutes after either IV or IP injections. Although both structures are within the blood-brain barrier (BBB), GM and WM revealed a steady decrease in T1 values at 24 and 72 hours after MnCl2 injection regardless of the route of administration. CONCLUSION: Systemic administration of MnCl2 by IV and IP routes induced similar time-course of T1 changes in different regions of the mouse brain. Acute effects of MnCl2 administration were mainly influenced by either the presence or absence of BBB. SC injection also provided significant T1 change at subacute stage after MnCl2 administration. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15779025&query_hl=1 ER - TY - JFULL T1 - Update of mortality and incidence rates for intrahepatic cholangiocarcinoma and other hepatobiliary tumours in England and Wales A1 - Khan, SA A1 - Mandeville, K A1 - Toledano, MB A1 - Thomas, HC A1 - Taylor-Robinson, SD J1 - J HEPATOL Y1 - 2005/04// VL - 42 SN - 0168-8278 SP - 95 EP - 95 ER - TY - JFULL T1 - Role of adhesion molecules in regulating neutrophil mobilisation from the bone marrow A1 - Rankin, SM A1 - Burdon, PCE A1 - Martin, C J1 - BRIT J HAEMATOL Y1 - 2005/04// VL - 129 SN - 0007-1048 SP - 43 EP - 43 ER - TY - JFULL T1 - Role of the RIP140 corepressor in ovulation and adipose biology. A1 - Steel, JH A1 - White, R A1 - Parker, MG J1 - J Endocrinol Y1 - 2005/04// VL - 185 SN - 0022-0795 SP - 1 EP - 9 N2 - RIP140 is a ligand-dependent corepressor for most, if not all, nuclear receptors. It is expressed widely in many different tissues, but the phenotype of mice devoid of RIP140 indicates that it plays a crucial role in the ovary and in adipose biology. Ovarian expression of RIP140 is cell-type-specific during follicular development and it is essential for oocyte release during ovulation, but not for luteinization of mature ovarian follicles. In adipose tissue, RIP140 is essential for normal fat accumulation and RIP140-null mice show decreased lipid storage even on a high-fat diet, with upregulation of mitochondrial uncoupling protein (UCP1) in some fat depots. Thus RIP140 plays a crucial role in female fertility and in energy homeostasis, and could be a target for infertility treatment, new contraceptive strategies or prevention of obesity. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15817822&query_hl=1 ER - TY - JFULL T1 - The CXC chemokine MIP-2 stimulates neutrophil mobilization from the rat bone marrow in a CD49d-dependent manner. A1 - Burdon, PC A1 - Martin, C A1 - Rankin, SM J1 - Blood Y1 - 2005/03/15/ VL - 105 SN - 0006-4971 SP - 2543 EP - 2548 N2 - The acute release of neutrophils from the bone marrow is a critical step in their trafficking to sites of inflammation. This process is stimulated by systemically acting inflammatory mediators, such as the CXC chemokines. In this study we have used a novel in situ perfusion system of the rat femoral bone marrow to directly investigate the role of specific adhesion molecules in chemokine-stimulated neutrophil mobilization. We show here that neutrophils mobilized in response to rat macrophage inflammatory protein-2 (MIP-2) shed L-selectin and expressed significantly higher levels of CD11b and CD49d. However, inhibition of L-selectin sheddase activity with KD-IX-73-4 had no effect on the number of neutrophils mobilized in response to rat MIP-2. Blockade of CD18, using a neutralizing monoclonal antibody (mAb), did not inhibit neutrophil mobilization but unexpectedly increased the rate and number of neutrophils released from the bone marrow in response to chemokine, suggesting that CD18 could play a role in neutrophil retention within the bone marrow. Blockade of CD49d using either a selective mAb or a specific antagonist resulted in a dramatic inhibition (> 75%) of the chemokine-stimulated neutrophil mobilization from the bone marrow. These data reveal contrasting roles for CD18 and CD49d in the retention and release of neutrophils from the bone marrow. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15542579&query_hl=1 ER - TY - JFULL T1 - Enhanced derivation of osteogenic cells from murine embryonic stem cells after treatment with ionic dissolution products of 58S bioactive sol-gel glass. A1 - Bielby, RC A1 - Pryce, RS A1 - Hench, LL A1 - Polak, JM J1 - Tissue Eng Y1 - 2005/03// VL - 11 SN - 1076-3279 SP - 479 EP - 488 N2 - Embryonic stem (ES) cells represent a potentially useful cell source for tissue regeneration. Previously, using factors known to enhance differentiation and mineralization of primary osteoblasts, we were able to generate cell populations enriched with osteoblasts from a murine ES cell source. Dexamethasone was a potent inducer of osteoblast differentiation and the timing of stimulation markedly increased the proportion of osteoblast lineage cells. This study examined whether inorganic stimuli derived from bioactive glasses could affect the differentiation of osteoblasts in an ES-cell based system. Previous work has demonstrated the ability of soluble ions released from bioactive glasses undergoing dissolution in vitro to stimulate gene expression characteristic of a mature phenotype in primary osteoblasts. We report here on the potential of soluble extracts prepared from 58S sol-gel bioactive glass to further enhance lineage-specific differentiation in murine ES cells. Differentiation of ES cells into osteogenic cells was characterized by the formation of multilayered, mineralized nodules. These nodules contained cells expressing the transcription factor runx2/cbfa-1, and deposition of osteocalcin in the extracellular matrix was detected by immunostaining. When differentiating cells were placed in an osteoblast maintenance medium supplemented with soluble extracts prepared from bioactive glass powders, we observed increased formation of mineralized nodules (98 +/- 6%, mean +/- SEM) and alkaline phosphatase activity (56 +/- 14%, mean +/- SEM) in a pattern characteristic of osteoblast differentiation. This effect of the glass extracts exhibited dose dependency, with alkaline phosphatase activity and nodule formation increasing with extract concentrations. Compared with medium supplemented with dexamethasone, which had previously been used to enhance osteoblast lineage derivation, the glass extracts were as effective at inducing formation of mineralized nodules by murine ES cells. When glass extracts were used in combination with dexamethasone, a further increase in the number of nodules was observed (110 +/- 16%; cf. 83 +/- 7% for dexamethasone alone). This study demonstrates the capacity of an entirely inorganic material to stimulate differentiation of ES cells toward a lineage with therapeutic potential in tissue-engineering applications. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15869426&query_hl=1 ER - TY - JFULL T1 - Effects of chronic administration of clenbuterol on function and metabolism of adult rat cardiac muscle. A1 - Soppa, GK A1 - Smolenski, RT A1 - Latif, N A1 - Yuen, AH A1 - Malik, A A1 - Karbowska, J A1 - Kochan, Z A1 - Terracciano, CM A1 - Yacoub, MH J1 - Am J Physiol Heart Circ Physiol Y1 - 2005/03// VL - 288 SN - 0363-6135 SP - H1468 EP - H1476 N2 - Clenbuterol (Clen), a beta(2)-agonist, is known to produce skeletal and myocardial hypertrophy. This compound has recently been used in combination with left ventricular assist devices for the treatment of end-stage heart failure to reverse or prevent the adverse effects of unloading-induced myocardial atrophy. However, the mechanisms of action of Clen on myocardial cells have not been fully elucidated. In an attempt to clarify this issue, we examined the effects of chronic administration of Clen on Ca(2+) handling and substrate preference in cardiac muscle. Rats were treated with either 2 mg x kg(-1) x day(-1) Clen or saline (Sal) for 4 wk with the use of osmotic minipumps. Ventricular myocytes were enzymatically dissociated. Cells were field stimulated at 0.5, 1, and 2 Hz, and cytoplasmic Ca(2+) transients were monitored with the use of the fluorescent indicator indo-1 acetoxymethyl ester. Two-dimensional surface area and action potentials in current clamp were also measured. We found that in the Clen group there was significant hypertrophy at the organ and cellular levels compared with Sal. In Clen myocytes, the amplitude of the indo-1 ratio transients was significantly increased. Sarcoplasmic reticulum Ca(2+) content, estimated by rapid application of 20 mM caffeine, was significantly increased in the Clen group. The action potential was prolonged in the Clen group compared with Sal. Carbohydrate contribution to the tricarboxylic cycle (Krebs cycle) flux was increased several times in the Clen group. This increase was associated with decreased expression of peroxisome proliferator-activated receptor-alpha. This study shows that chronic administration of Clen induces cellular hypertrophy and increases oxidative carbohydrate utilization together with an increase in sarcoplasmic reticulum Ca(2+) content, which results in increased amplitude of the Ca(2+) transients. These effects could be important when Clen is used in conjunction with left ventricular assist devices treatment. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15528231&query_hl=1 ER - TY - JFULL T1 - Effects of Lovastatin on the proliferation and cell cycle of rat cardiac fibroblasts induced by aldosterone in vitro A1 - Cui, W A1 - Fang, HP A1 - Shan, B A1 - Hao, YM A1 - Du, J J1 - J AM COLL CARDIOL Y1 - 2005/02/01/ VL - 45 SN - 0735-1097 SP - 394A EP - 394A ER - TY - JFULL T1 - Hey1, a mediator of notch signaling, is an androgen receptor corepressor. A1 - Belandia, B A1 - Powell, SM A1 - García-Pedrero, JM A1 - Walker, MM A1 - Bevan, CL A1 - Parker, MG J1 - Mol Cell Biol Y1 - 2005/02// VL - 25 SN - 0270-7306 SP - 1425 EP - 1436 N2 - Hey1 is a member of the basic helix-loop-helix-Orange family of transcriptional repressors that mediate Notch signaling. Here we show that transcription from androgen-dependent target genes is inhibited by Hey1 and that expression of a constitutively active form of Notch is capable of repressing transactivation by the endogenous androgen receptor (AR). Our results indicate that Hey1 functions as a corepressor for AF1 in the AR, providing a mechanism for cross talk between Notch and androgen-signaling pathways. Hey1 colocalizes with AR in the epithelia of patients with benign prostatic hyperplasia, where it is found in both the cytoplasm and the nucleus. In marked contrast, we demonstrate that Hey1 is excluded from the nucleus in most human prostate cancers, raising the possibility that an abnormal Hey1 subcellular distribution may have a role in the aberrant hormonal responses observed in prostate cancer. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15684393&query_hl=1 ER - TY - JFULL T1 - T:G mismatch-specific thymine-DNA glycosylase (TDG) as a coregulator of transcription interacts with SRC1 family members through a novel tyrosine repeat motif. A1 - Lucey, MJ A1 - Chen, D A1 - Lopez-Garcia, J A1 - Hart, SM A1 - Phoenix, F A1 - Al-Jehani, R A1 - Alao, JP A1 - White, R A1 - Kindle, KB A1 - Losson, R A1 - Chambon, P A1 - Parker, MG A1 - Schär, P A1 - Heery, DM A1 - Buluwela, L A1 - Ali, S J1 - Nucleic Acids Res Y1 - 2005/// VL - 33 SN - 1362-4962 SP - 6393 EP - 6404 N2 - Gene activation involves protein complexes with diverse enzymatic activities, some of which are involved in chromatin modification. We have shown previously that the base excision repair enzyme thymine DNA glycosylase (TDG) acts as a potent coactivator for estrogen receptor-alpha. To further understand how TDG acts in this context, we studied its interaction with known coactivators of nuclear receptors. We find that TDG interacts in vitro and in vivo with the p160 coactivator SRC1, with the interaction being mediated by a previously undescribed motif encoding four equally spaced tyrosine residues in TDG, each tyrosine being separated by three amino acids. This is found to interact with two motifs in SRC1 also containing tyrosine residues separated by three amino acids. Site-directed mutagenesis shows that the tyrosines encoded in these motifs are critical for the interaction. The related p160 protein TIF2 does not interact with TDG and has the altered sequence, F-X-X-X-Y, at the equivalent positions relative to SRC1. Substitution of the phenylalanines to tyrosines is sufficient to bring about interaction of TIF2 with TDG. These findings highlight a new protein-protein interaction motif based on Y-X-X-X-Y and provide new insight into the interaction of diverse proteins in coactivator complexes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16282588&query_hl=1 ER - TY - JFULL T1 - Sarcolemmal L-type calcium channel : Intracellular calcium release channel (ryanodine receptor) stoichiometry is reduced in pressure overload heart failure but not in chronic catecholaminergic exposure. A1 - Scoote, M A1 - MacLeod, KT A1 - Harding, SE A1 - Kingsbury, MP A1 - Turner, MA A1 - Sheridan, DJ A1 - Williams, AJ J1 - BIOPHYS J Y1 - 2005/01// VL - 88 SN - 0006-3495 SP - 487A EP - 488A ER - TY - JFULL T1 - Distribution of sodium channels in ventricular heart cells A1 - Duclohier, HPM A1 - Gorelik, J A1 - Yang, LQ A1 - Harding, SE A1 - Korchev, YE J1 - BIOPHYS J Y1 - 2005/01// VL - 88 SN - 0006-3495 SP - 378A EP - 378A ER - TY - JFULL T1 - In situ monitoring of chondrocyte response to bioactive scaffolds using Raman spectroscopy A1 - Jones, JR A1 - Vats, A A1 - Notingher, L A1 - Gough, JE A1 - Tolley, NS A1 - Polak, JM A1 - Hench, LL J1 - KEY ENG MAT Y1 - 2005/// VL - 284-286 SP - 623 EP - 626 N2 - Septal cartilage is widely used for the repair of soft tissue defects in the head, neck and nose. Tissue Engineering techniques are being investigated to create cartilage in vitro by seeding appropriate cells on resorbable scaffolds. In this study, human chondrocytes were cultured on macroporous bioactive glass foam scaffolds. The aim was to investigate how Raman spectroscopy could be used as a non-invasive technique to monitor the response of chondrocytes to a 3D scaffold in real time. The spectra were compared to scanning electron microscope (SEM) micrographs and immunohistochemistry results. ER - TY - JFULL T1 - Antisense strategies for treatment of heart failure. A1 - Harding, SE A1 - del Monte, F A1 - Hajjar, RJ J1 - Methods Mol Med Y1 - 2005/// VL - 106 SN - 1543-1894 SP - 69 EP - 82 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15375313&query_hl=1 ER - TY - JFULL T1 - Raman spectroscopy: Potential tool for in situ characterization of bone cell differentiation A1 - Notingher I A1 - Jell G A1 - Notingher PL A1 - Bisson I A1 - Polak JM A1 - Hench LL J1 - Key Engineering Materials Y1 - 2005/// SP - 545 EP - 548 ER - TY - JFULL T1 - Overexpression of Na/K ATPase alpha-1 and 2 subunits differentially affects calcium regulation in rat ventricular myocytes A1 - Malik, AH A1 - Khan, SG A1 - Harding, SE A1 - Terracciano, CMN J1 - BIOPHYS J Y1 - 2005/01// VL - 88 SN - 0006-3495 SP - 297A EP - 297A ER - TY - JFULL T1 - Multipath source routing in sensor networks based on route ranking A1 - Huang, C A1 - Chatterjee, M A1 - Cui, W A1 - Guha, R J1 - LECT NOTES COMPUT SC Y1 - 2005/// VL - 3741 SN - 0302-9743 SP - 99 EP - 104 N2 - Multipath source routing is an effective way to exploit the redundant routes that are usually common in dense sensor networks. In this paper, we present a multipath source routing algorithm that uses a ranking technique to distinguish between the quality of different routes for the same source-destination pair. A ranking coefficient is calculated for each route based on three different metrics- energy, delay and reliability. The number of parallel routes that is considered is governed by the minimum reliability requirements. Simulation experiments are conducted that show that multipath routing can increase the reliability, and dissipate energy more evenly among the nodes. ER - TY - JFULL T1 - Pantoprazole in severe acid-peptic disease: the effectiveness and safety of 5 years' continuous treatment A1 - Bardhan, KD A1 - Bishop, AE A1 - Polak, JM A1 - Romanska, HM A1 - Rowland, A A1 - Thompson, M A1 - Morris, P A1 - Schaefer-Preuss, S A1 - Luehmann, R A1 - McCaldin, B J1 - DIGEST LIVER DIS Y1 - 2005/01// VL - 37 SN - 1590-8658 SP - 10 EP - 22 N2 - Objective. This is our final report on the clinical effectiveness and safety of long-term pantoprazole in patients with severe peptic ulcer or reflux disease during continuous treatment for up to 5 years.Methods. Patients (n = 150) with peptic ulcer or reflux erosive oesophagitis running an aggressive course or with complications, and refractory to H2-receptor antagonists, were entered into this 5-year programme. Assessment was by serial endoscopy, clinical examination, serum gastrin estimation, gastric mucosal histology and mucosal endocrine cell quantification.Results. Healing results were presented earlier. The estimated rates of remission on maintenance treatment with pantoprazole (n=115) were 82% at 1 year, 75% at 2 years, 72% at 3 years, 70% at 4 years and 68% at 5 years. Helicobacter pylori infection appeared not to influence the outcome in reflux patients, with roughly two-thirds continuing in remission irrespective of infection. Only four patients had adverse events considered to be definitely related to pantoprazole. Median gastrin levels rose by 1.5-2-fold and were higher in those with H. pylori infection; 13 patients had levels >500 ng/L on at least one occasion, but these high levels were not sustained. Histological changes were more marked in patients infected with H. pylori: chronic gastritis decreased in the antrum and increased in the corpus, which also showed atrophic changes. The total number of endocrine cells in the antrum showed little variation over 60 months but fell by around one-third in the corpus.Conclusion. Long-term treatment with pantoprazole is effective and safe. (C) 2004 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved. ER - TY - JFULL T1 - Stably transfected human embryonic stem cell clones express OCT4-specific green fluorescent protein and maintain self-renewal and pluripotency. A1 - Gerrard, L A1 - Zhao, D A1 - Clark, AJ A1 - Cui, W J1 - Stem Cells Y1 - 2005/// VL - 23 SN - 1066-5099 SP - 124 EP - 133 N2 - Human embryonic stem cells (hESCs) are derived from the inner cell mass of preimplantation embryos; they can be cultured indefinitely and differentiated into many cell types in vitro. These cells therefore have the ability to provide insights into human disease and provide a potential unlimited supply of cells for cell-based therapy. Little is known about the factors that are important for maintaining undifferentiated hESCs in vitro, however. As a tool to investigate these factors, transfected hES clonal cell lines were generated; these lines are able to express the enhanced green fluorescent protein (EGFP) reporter gene under control of the OCT4 promoter. OCT4 is an important marker of the undifferentiated state and a central regulator of pluripotency in ES cells. These OCT4-EGFP clonal cell lines exhibit features similar to parental hESCs, are pluripotent, and are able to produce all three embryonic germ layer cells. Expression of OCT4-EGFP is colocalized with endogenous OCT4, as well as the hESC surface antigens SSEA4 and Tra-1-60. In addition, the expression is retained in culture for an extensive period of time. Differentiation of these cells toward the neural lineage and targeted knockdown of endogenous OCT4 expression by RNA interference downregulated the EGFP expression in these cell lines, and this correlates closely with the reduction of endogenous OCT4 expression. Therefore, these cell lines provide an easy and noninvasive method to monitor expression of OCT4 in hESCs, and they will be invaluable for studying not only OCT4 function in hESC self-renewal and differentiation but also the factors required for maintenance of undifferentiated hESCs in culture. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15625129&query_hl=1 ER - TY - JFULL T1 - Less Na+/H+-exchanger to treat heart failure: a simple solution for a complex problem? A1 - Stagg, MA A1 - Terracciano, CM J1 - Cardiovasc Res Y1 - 2005/01/01/ VL - 65 SN - 0008-6363 SP - 10 EP - 12 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15621028&query_hl=1 ER - TY - JFULL T1 - Gene therapy progress and prospects: In tissue engineering A1 - Polak J A1 - Hench L J1 - Gene Therapy Y1 - 2005/// SP - 1 EP - 9 ER - TY - JFULL T1 - A dynamic switch in the replication timing of key regulator genes in embryonic stem cells upon neural induction. A1 - Perry, P A1 - Sauer, S A1 - Billon, N A1 - Richardson, WD A1 - Spivakov, M A1 - Warnes, G A1 - Livesey, FJ A1 - Merkenschlager, M A1 - Fisher, AG A1 - Azuara, V J1 - Cell Cycle Y1 - 2004/12// VL - 3 SN - 1551-4005 SP - 1645 EP - 1650 N2 - Mammalian embryonic stem (ES) cells can either self-renew or generate progenitor cells that have a more restricted developmental potential. This provides an important model system to ask how pluripotency, cell commitment and differentiation are regulated at the level of chromatin-based changes that distinguish stem cells from their differentiated progeny. Here we show that the differentiation of ES cells to neural progenitors results in dynamic changes in the epigenetic status of multiple genes that encode transcription factors critical for early embryonic development or lineage specification. In particular, we demonstrate that DNA replication at a subset of neural-associated genes including Pax3, Pax6, Irx3, Nkx2.9 and Mash1 is advanced upon neural induction, consistent with increased locus accessibility. Conversely, many ES-associated genes including Oct4, Nanog, Utf1, Foxd3, Cripto and Rex1 that replicate early in ES cells switch their replication timing to later in S-phase in response to differentiation. Detailed analysis of the Rex1 locus reveals that delayed replication extends to a 2.8 Mb region surrounding the gene and is associated with substantial reductions in the level of histone H3K9 and H4 acetylation at the promoter. These results show that loss of pluripotency (and lineage choice) is associated with extensive and predictable changes in the replication timing of key regulator genes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15611653&query_hl=1 ER - TY - JFULL T1 - Expression of beta-catenin gene in human hematopoietic cell lines and leukemia patients. A1 - Qiu, LG A1 - Mai, YJ A1 - Yu, Z A1 - Wang, YF A1 - Zhou, Y A1 - Cui, W J1 - BLOOD Y1 - 2004/11/16/ VL - 104 SN - 0006-4971 SP - 168B EP - 168B ER - TY - JFULL T1 - Depletion of the CD1d-restricted NKT cells suppresses in vitro alloreactivity: A possible means to prevent aGVHD. A1 - Pattersons, S A1 - Kotsianidis, I A1 - Almeida, A A1 - Politou, M A1 - David, R A1 - Rahemtulla, A A1 - Cerundolo, V A1 - Roberts, I A1 - Karadimitris, A J1 - BLOOD Y1 - 2004/11/16/ VL - 104 SN - 0006-4971 SP - 838A EP - 839A ER - TY - JFULL T1 - Human CD4 negative NKT cells potently suppress alloreactive responses in a TGF-beta and IL-10-Independent manner A1 - Patterson, S A1 - Kotsiandis, I A1 - Almeida, A A1 - Politou, M A1 - Rahemtulla, A A1 - Cerundolo, V A1 - Roberts, I A1 - Karadimitris, A J1 - BLOOD Y1 - 2004/11/16/ VL - 104 SN - 0006-4971 SP - 131A EP - 131A ER - TY - JFULL T1 - Evidence that human NKT cells enhance haemopoiesis through recognition of CD1d expressed in haemopoietic stem cells with long term clonogenic capacity A1 - Kotsianidis, L A1 - Patterson, S A1 - Politou, M A1 - Almeida, A A1 - Pantelidou, D A1 - Tsatalas, C A1 - Bourikas, G A1 - Roberts, I A1 - Karadimitris, A J1 - BLOOD Y1 - 2004/11/16/ VL - 104 SN - 0006-4971 SP - 119B EP - 119B ER - TY - JFULL T1 - Targeted retroviral transduction of c-kit+ hematopoietic cells using novel ligand display technology. A1 - Chandrashekran, A A1 - Gordon, MY A1 - Casimir, C J1 - Blood Y1 - 2004/11/01/ VL - 104 SN - 0006-4971 SP - 2697 EP - 2703 N2 - Gene therapy for a wide variety of disorders would be greatly enhanced by the development of vectors that could be targeted for gene delivery to specific populations of cells. We describe here high-efficiency targeted transduction based on a novel targeting strategy that exploits the ability of retroviruses to incorporate host cell proteins into the surface of the viral particle as they bud through the plasma membrane. Ecotropic retroviral particles produced in cells engineered to express the membrane-bound form of stem cell factor (mbSCF) transduce both human cell lines and primary cells with high efficiency in a strictly c-kit (SCF receptor)-dependent fashion. The availability of efficient targeted vectors provides a platform for the development of a new generation of therapies using in vivo gene delivery. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15256424&query_hl=1 ER - TY - JFULL T1 - P13-kinase signalling pathway is involved in adipogenic differentiation of fetal liver derived mesenchymal stem cells A1 - Taribagil, SS A1 - Chandrashekran, A A1 - Stamp, G A1 - Gordon, MY A1 - Lalani, E J1 - MOL BIOL CELL Y1 - 2004/11// VL - 15 SN - 1059-1524 SP - 338A EP - 338A ER - TY - JFULL T1 - Apoptosis--a significant cause of bone cell death in osteonecrosis of the femoral head. A1 - Calder, JD A1 - Buttery, L A1 - Revell, PA A1 - Pearse, M A1 - Polak, JM J1 - J Bone Joint Surg Br Y1 - 2004/11// VL - 86 SN - 0301-620X SP - 1209 EP - 1213 N2 - Osteonecrosis of the femoral head usually affects young individuals and is responsible for up to 12% of total hip arthroplasties. The underlying pathophysiology of the death of the bone cells remains uncertain. We have investigated nitric oxide mediated apoptosis as a potential mechanism and found that steroid- and alcohol-induced osteonecrosis is accompanied by widespread apoptosis of osteoblasts and osteocytes. Certain drugs or their metabolites may have a direct cytotoxic effect on cancellous bone of the femoral head leading to apoptosis rather than purely necrosis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15568539&query_hl=1 ER - TY - JFULL T1 - An HSP90-mimic peptide revealed by fingerprinting the pool of antibodies from ovarian cancer patients A1 - Vidal, CI A1 - Mintz, PJ A1 - Lu, K A1 - Ellis, LM A1 - Manenti, L A1 - Giavazzi, R A1 - Gershenson, DM A1 - Broaddus, R A1 - Liu, J A1 - Arap, W A1 - Pasqualini, R J1 - Oncogene Y1 - 2004/11// IS - 55 VL - 23 SN - 0950-9232 SP - 8859 EP - 8867 ER - TY - JFULL T1 - Growth factor displayed on the surface of retroviral particles without manipulation of envelope proteins is biologically active and can enhance transduction. A1 - Chandrashekran, A A1 - Gordon, MY A1 - Darling, D A1 - Farzaneh, F A1 - Casimir, C J1 - J Gene Med Y1 - 2004/11// VL - 6 SN - 1099-498X SP - 1189 EP - 1196 N2 - BACKGROUND: The therapeutic potential of retroviruses can be significantly enhanced by display of specific molecules on the retroviral surface. This has been conventionally achieved by the manipulation of retroviral envelope proteins. In this report we have tested whether the natural budding mechanism of the retrovirus could be exploited to incorporate a specific molecule into the retroviral surface. METHODS: Retroviral packaging cells were engineered to express the membrane-bound form of human stem cell factor (mbSCF). Surface expression of mbSCF on retroviral packaging cells was confirmed by immunofluorescence and flow cytometry. Incorporation of mbSCF into retroviral particles was demonstrated by virus-binding assay and immunomagnetic capture of virus using antibody to SCF. Retroviral supernatants were tested for activity of the incorporated cytokine by proliferation assays on factor-dependent cells. Amphotropic retrovirus displaying surface mbSCF was used to transduce SCF receptor-positive haematopoietic cells. RESULTS: Retroviruses incorporating surface SCF showed increased levels of binding to cells (MO7e) expressing the SCF receptor, c-kit. mbSCF displayed on the viral surface retained levels of biological activity comparable with those of soluble recombinant growth factor. Transduction of c-kit-positive target cells with viruses displaying mbSCF showed enhanced levels of transduction in comparison with unmodified viruses. CONCLUSIONS: Expression of the membrane-bound form of human stem cell factor (mbSCF) on the surface of retroviral packaging cells allows its efficient incorporation into retrovirus particles in a biologically active form, opening up the possibility for the use of retroviral display in many therapeutic areas, such as in gene therapy, drug delivery and in the development of novel vaccines. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15468194&query_hl=1 ER - TY - JFULL T1 - Beta-adrenoceptor subtype dependence of chronotropy in mouse embryonic stem cell-derived cardiomyocytes. A1 - Ali, NN A1 - Xu, X A1 - Brito-Martins, M A1 - Poole-Wilson, PA A1 - Harding, SE A1 - Fuller, SJ J1 - Basic Res Cardiol Y1 - 2004/11// VL - 99 SN - 0300-8428 SP - 382 EP - 391 N2 - Cardiomyocytes derived from embryonic stem cells (ESCM) have potential both as an experimental model for investigating cardiac physiology and as a source for tissue repair. For both reasons it is important to characterise the responses of these cells, and one of the key modulators of contraction is the beta-adrenergic system. We therefore undertook a detailed study of the response of the spontaneous beating rate of ESCM to beta-adrenoceptor (betaAR) stimulation. Embryoid bodies (EBs) were generated from murine ES line E14Tg2a by the hanging drop method, followed by plating. Spontaneously beating areas were seen starting from 9-14 days after differentiation: the experiments described here were performed on EBs between developmental day 19 and 48. Beating cell layers were seeded with charcoal to allow tracking of movement by a video-edge detection system. Experiments were performed in physiological medium containing 1 mM Ca2+ at 37 degrees C. Isoprenaline (Iso) increased beating rate with an EC50 value of 52 nM. Iso (0.3 microM) increased basal rate from 67 +/- 7 beats per minute (bpm) to 138 +/- 18 bpm, P < 0.001, n = 22. At earlier developmental time points the response to Iso was not maintained through 5 min exposure; this spontaneous desensitisation only being observed before day 36. A repeat application of Iso after a wash period of 20 min produced reproducible effects on beating rate. Subtype dependence of the betaAR response was determined by comparing an initial response with a second in the presence of selective beta1- or beta2AR antagonists. In the presence of the specific beta1AR-blocker CGP 20712A (300 nM) the increase in rate with Iso was reduced from 207 +/- 42% of basal to 128 +/- 13%, P < 0.01. With the beta2AR-blocker ICI 118,551 (50 nM) there was no significant change in Iso response. Exposure to the muscarinic agonist, carbachol (10 microM), inhibited the increase in frequency mediated by isoprenaline, but had mixed stimulatory and inhibitory effects on basal rate. This study extends the characterisation of ESCM as a preparation for studying receptor pharmacology, and indicates that the beta1AR is the predominant subtype mediating increases in contraction rate in murine ESCM. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15365729&query_hl=1 ER - TY - JFULL T1 - p38-MAPK signaling pathway plays an important role in the acute negative inotropic effect of beta-blockers in human ventricular myocytes A1 - Zheng, ZL A1 - Gong, HB A1 - Yang, LQ A1 - Petrou, M A1 - Harding, SE J1 - CIRCULATION Y1 - 2004/10/26/ VL - 110 SN - 0009-7322 SP - 91 EP - 92 ER - TY - JFULL T1 - Connexin43 overexpression increases gap junctional conductance between skeletal myoblasts and adult rat ventricular myocytes in coculture A1 - Stagg, MA A1 - Coppen, SR A1 - Lee, A A1 - Varela-Carver, A A1 - Brand, NJ A1 - Fukushima, S A1 - Suzuki, K A1 - Terracciano, CM J1 - CIRCULATION Y1 - 2004/10/26/ VL - 110 SN - 0009-7322 SP - 280 EP - 280 ER - TY - JFULL T1 - An investigation of gap junction formation between skeletal myoblasts and adult cardiac myocytes using myoblasts overexpressing Connexin43 A1 - Coppen, SR A1 - Varela-Carver, A A1 - Stagg, MA A1 - Lee, J A1 - Fukushima, S A1 - Brand, NJ A1 - Terracciano, CM A1 - Suzuki, K J1 - CIRCULATION Y1 - 2004/10/26/ VL - 110 SN - 0009-7322 SP - 396 EP - 396 ER - TY - JFULL T1 - Chondrogenic differentiation of murine embryonic stem cells: effects of culture conditions and dexamethasone. A1 - Tanaka, H A1 - Murphy, CL A1 - Murphy, C A1 - Kimura, M A1 - Kawai, S A1 - Polak, JM J1 - J Cell Biochem Y1 - 2004/10/15/ VL - 93 SN - 0730-2312 SP - 454 EP - 462 N2 - Pluripotent embryonic stem (ES) cells have the capability to differentiate to various cell types and may represent an alternative cell source for the treatment of cartilage defects. Here, we show that differentiation of ES cells toward the chondrogenic lineage can be enhanced by altering the culture conditions. Chondrogenesis was observed in intact embryoid body (EB) cultures, as detected by an increase in mRNA levels for aggrecan and Sox9 genes. Collagen IIB mRNA, the mature chondrocyte-specific splice variant, was absent at day 5, but appeared at later time points. Dexamethasone treatment of alginate-encapsulated EB cultures did not have a strong chondrogenic effect. Nor was chondrogenesis enhanced by alginate encapsulation compared to simple plating of EBs. However, disruption of day 5 EBs and culture as a micromass or pelleted mass, significantly enhanced the expression of the cartilage marker gene collagen type II and the transcription factor Sox9 compared to all other treatments. Histological and immunohistochemical analysis of pellet cultures revealed cartilage-like tissue characterized by metachromatically stained extracellular matrix and type II collagen immunoreactivity, indicative of chondrogenesis. These findings have potentially important implications for cartilage tissue engineering, since they may enable the increase in differentiated cell numbers needed for the in vitro development of functional cartilaginous tissue suitable for implantation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15372628&query_hl=1 ER - TY - JFULL T1 - CD44 intracellular domain stimulates osteoclastogenesis in a presenilin-dependent manner. A1 - Cui, W A1 - Chalouni, C A1 - Zhang, J A1 - Ke, H A1 - Vignery, A J1 - J BONE MINER RES Y1 - 2004/10// VL - 19 SN - 0884-0431 SP - S82 EP - S82 ER - TY - JFULL T1 - Effect of overexpressed adenylyl cyclase VI on beta(1)- and beta(2)-adrenoceptor responses in adult rat ventricular myocytes A1 - Stark, JCC A1 - Haydock, SF A1 - Foo, R A1 - Brown, MJ A1 - Harding, SE J1 - BRIT J PHARMACOL Y1 - 2004/10// VL - 143 SN - 0007-1188 SP - 465 EP - 476 N2 - 1 Adenylyl cyclase VI (ACVI) is one of the most abundantly expressed beta adrenergic receptor (betaAR)coupled cyclases responsible for cyclic AMP ( cAMP) production within the mammalian myocardium. We investigated the role of ACVI in the regulation of cardiomyocyte contractility and whether it is functionally coupled with beta(1) adrenergic receptor (beta(1)AR).2 Recombinant adenoviruses were generated for ACVI and for antisense to ACVI ( AS). Adult rat ventricular myocytes were transfected with ACVI virus, AS or both ( SAS). Adenovirus for green fluorescent protein (GFP) served as control. Myocyte contraction amplitudes (% shortening) and relaxation times (R50) were analysed. ACVI function was determined using cAMP assays.3 ACVI-transfected cells demonstrated a strong 139 kDa ACVI protein band compared to controls. ACVI myocytes had higher steady-state intracellular cAMP levels than GFP myocytes when unstimulated ( GFP vs ACVI = 6.60 +/- 0.98 vs 14.2 +/- 2.1 fmol cAMP/viable cell, n = 4, P<0.05) and in the presence of 1 μM isoprenaline or 10 μM forskolin.4 ACVI myocytes had increased basal contraction (% shortening: GFP vs ACVI: 1.90 +/- 1.36 vs 3.91 +/- 2.29, P<0.0001) and decreased basal R50 (GFP vs ACVI: 62.6 +/- 24.2 ms ( n = 50) vs 45.0 +/- 17.2 ms ( n = 248), P<0.0001). ACVI myocyte responses were increased for forskolin (E-max: GFP = 6.70 +/- 1.59 ( n = 6); ACVI = 9.06 +/- 0.69 ( n = 14), P<0.01) but not isoprenaline.5 ACVI myocyte responses were increased (E-max: GFP vs ACVI = 3.16 +/- 0.77 vs 5.10 +/- 0.60, P<0.0001) to xamoterol ( a partial β(1)AR-selective agonist) under β(2)AR blockade (+50 nM ICI 118, 551). AS decreased both control and ACVI-stimulated xamoterol responses (E-max: AS = 2.59 +/- 1.42, SAS = 1.38 +/- 0.5). ACVI response was not mimicked by IBMX. Conversely, response through β(2) adrenergic receptor (β(2)AR) was decreased in ACVI myocytes.6 In conclusion, ACVI overexpression constitutively increases myocyte contraction amplitudes by raising cAMP levels. Native ACVI did not contribute to basal cAMP production or contraction amplitude and only to a minor extent to the forskolin response. β(1)AR but not β(2)AR coupling was dependent on ACVI. ER - TY - JFULL T1 - Cell surface expression of the stress response chaperone GRP78 enables tumor targeting by circulating ligands A1 - Arap, MA A1 - Lahdenranta, J A1 - Mintz, PJ A1 - Hajitou, A A1 - Sarkis, AS A1 - Arap, W A1 - Pasqualini, R J1 - Cancer Cell Y1 - 2004/09// IS - 3 VL - 6 SN - 1535-6108 SP - 275 EP - 284 ER - TY - JFULL T1 - In vitro differentiation and in vivo mineralization of osteogenic cells derived from human embryonic stem cells. A1 - Bielby, RC A1 - Boccaccini, AR A1 - Polak, JM A1 - Buttery, LD J1 - Tissue Eng Y1 - 2004/09// VL - 10 SN - 1076-3279 SP - 1518 EP - 1525 N2 - The first report of the derivation of embryonic stem (ES) cell lines from human blastocysts had major implications for research into developmental biology and regenerative medicine. Finding efficient and reproducible methods to derive therapeutically useful cells from an ES cell source is a key feature of many regenerative medicine strategies. We have previously demonstrated that it is possible to induce osteogenic differentiation of murine ES cells by supplementing the culture medium with ascorbic acid, beta-glycerophosphate, and dexamethasone. This study investigated whether methods for driving osteogenic differentiation developed with murine ES cells could be applied successfully to human ES cells. The H1 line was propagated in vitro on murine feeder layers and shown to be pluripotent by expression of the markers Oct-4 and SSEA-4. Subsequently, differentiation was initiated via embryoid body (EB) formation and, after 5 days in suspension culture, cells harvested from EBs were replated in a medium containing osteogenic supplements. We found that the treatment regimen previously identified as optimal for murine ES cells, and in particular the addition of dexamethasone at specific time points, also induced the greatest osteogenic response from human ES cells. We identified mineralizing cells in vitro that immunostained positively for osteocalcin and found an increase in expression of an essential bone transcription factor, Runx2. When implanted into SCID mice on a poly-D, L-lactide (PDLLA) scaffold, the cells had the capacity to give rise to mineralized tissue in vivo. After 35 days of implantation, regions of mineralized tissue could be identified within the scaffold by von Kossa staining and immunoexpression of the human form of osteocalcin. We did not see any evidence of teratoma formation. These data therefore demonstrate the derivation of osteoblasts from pluripotent human ES cells with the capacity to form mineralized tissue both in vitro and in vivo. We have also shown that a culture methodology established for differentiation of murine ES cells was entirely transferable to human ES cells. Further development of this technology will result in the capacity to generate sufficient yields of osteogenic cells for use in skeletal tissue repair. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15588411&query_hl=1 ER - TY - JFULL T1 - Differentiation of osteoblasts from murine embryonic stem cells by overexpression of the transcriptional factor osterix. A1 - Tai, G A1 - Polak, JM A1 - Bishop, AE A1 - Christodoulou, I A1 - Buttery, LD J1 - Tissue Eng Y1 - 2004/09// VL - 10 SN - 1076-3279 SP - 1456 EP - 1466 N2 - Osterix is a transcription factor crucial for the normal development of the osteoblast. Here we have investigated whether the osteogenic differentiation of murine embryonic stem (ES) cells can be induced by overexpression of osterix. Differentiation was initiated by formation of embryoid bodies (EB) which were then dispersed and cultured in alpha-minimum essential medium supplemented with L-ascorbate phosphate and alpha-glycerophosphate for up to 21 days. osterix was found to induce expression of several osteoblast-specific markers, as confirmed by immunostaining and real-time RT-PCR. The expression of genes encoding osteocalcin and Cbfa1 was upregulated and the formation of mineralized bone nodules was significantly increased by osterix transfection. In combination with dexamethasone, bone nodule formation was further increased in osterix-transfected cells. Expression of both Sox-9 and PPAR-gamma, genes that are associated with chondrocyte and adipocyte differentiation, was initially increased in the osterix-transfected cells but was downregulated after day 7. This suggests that the process of osterix-induced differentiation of ES cells involves transition through an intermediate bi- or tripotential progenitor cell population. In conclusion, this cell differentiation strategy is useful not only for generating osteoblastic cells from ES cells, but also for investigating factors that influence this process and potentially delineating the ontogeny of the osteoblast. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15588405&query_hl=1 ER - TY - JFULL T1 - The effects of overexpression of the Na+/Ca2+ exchanger on calcium regulation in hypertrophied mouse cardiac myocytes. A1 - Stagg, MA A1 - Malik, AH A1 - MacLeod, KT A1 - Terracciano, CM J1 - Cell Calcium Y1 - 2004/08// VL - 36 SN - 0143-4160 SP - 111 EP - 118 N2 - In cardiac hypertrophy and failure it has been shown that the amount of Na/Ca exchanger protein can increase. Several studies have investigated this modification in overt heart failure. However, the role of Na/Ca exchanger overexpression during the development of hypertrophy is unknown. To address this question we investigated Ca2+ regulation in an early stage of cardiac hypertrophy before signs of heart failure occurred and evaluated the role of Na/Ca exchanger overexpression. Cardiac hypertrophy was induced by a constant infusion of angiotensin II (Ang, 1 microg/min/kg) via an osmotic pump for 14 days. Thereafter, ventricular myocytes from either wild type (NON) or transgenic mice overexpressing the Na/Ca exchanger (TR) were isolated. Myocytes were loaded with indo-1 AM or fluo-4 AM to monitor cytoplasmic [Ca2+] with all experiments performed at 37 degrees C. In myocytes exposed to Ang there was an increase in cell capacitance of more than 20% indicating cellular hypertrophy. Ca2+ transients were prolonged in hypertrophied NON myocytes but not in TR myocytes. Action potentials had a less negative plateau in TR myocytes. Sarcoplasmic reticulum (SR) Ca2+ content, measured using rapid caffeine application, was greater in TR myocytes but unaffected by hypertrophy. Ca2+ spark frequency was significantly greater in TR. Na/Ca exchanger overexpression prevented the prolongation of the Ca2+ transient observed in hypertrophy and maintained a similar SR Ca2+ leak suggesting a compensatory role in Ca2+ regulation in hypertrophied cardiac myocytes from transgenic mice. We suggest this compensatory effect is mediated by increased SR Ca2+ content and faster Ca2+ removal via the Na/Ca exchanger. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15193859&query_hl=1 ER - TY - JFULL T1 - Acute negative inotropic effect of beta(2)AR-blockers through p38-MAPK signaling pathway in human ventricular myocytes A1 - Zheng, Z A1 - Gong, H A1 - Petrou, M A1 - Yang, LQ A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2004/07// VL - 37 SN - 0022-2828 SP - 297 EP - 298 ER - TY - JFULL T1 - Time- and concentration-dependent effects of dissolution products of 58S sol-gel bioactive glass on proliferation and differentiation of murine and human osteoblasts. A1 - Bielby, RC A1 - Christodoulou, IS A1 - Pryce, RS A1 - Radford, WJ A1 - Hench, LL A1 - Polak, JM J1 - Tissue Eng Y1 - 2004/07// VL - 10 SN - 1076-3279 SP - 1018 EP - 1026 N2 - Bone loss is a significant clinical problem, and treatments utilizing donated graft material are limited. To meet future demands in the healthcare industry, there has been a shift of outlook toward the use of bioactive materials for tissue regeneration. A number of in vivo and in vitro studies have highlighted the potential of the bioactive glass ceramic 45S5 Bioglass as a synthetic regenerative scaffold. The application of sol-gel processing techniques has led to the synthesis of mesoporous bioactive glasses with greater textural and compositional variety. In this study, we evaluated the effects of supplemented tissue culture medium containing up to 203 ppm silica prepared by static soaking of particles of 58S sol-gel bioactive glass (58% SiO(2), 33% CaO, 9% P(2)O(5)) on the in vitro proliferation and differentiation of murine and human primary osteoblasts. These extracts had a higher silica content than those used previously in studies of 45S5 Bioglass, because of the faster rates of ion exchange permitted by the higher surface area-to-volume ratio of mesoporous glass. We found that osteoblasts from both species increased their proliferation in response to the glass-conditioned medium. In addition, the extent to which supplemented medium could alter cell differentiation varied with time in culture. Proliferation induced by supplemented medium paralleled effects induced by treatment with basic fibroblast growth factor, a known mitogenic growth factor for osteoblasts. Bone nodule formation was also increased by exposure to the glass-conditioned medium and this effect was positively correlated with the dose of glass used to prepare the medium. Apoptosis was stimulated by glass-conditioned medium in murine osteoblasts, but inhibited in human osteoblasts. These data demonstrate the bioactive effects of dissolution products derived from sol-gel materials on primary osteoblasts and complements in vivo studies that indicate the suitability of this material as a bone graft substitute. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15363159&query_hl=1 ER - TY - JFULL T1 - Does beta-blocker-mediated stimulation of gi contribute to recovery mechanisms? A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2004/07// VL - 37 SN - 0022-2828 SP - 324 EP - 325 ER - TY - JFULL T1 - beta-adrenoceptor blockers as agonists: coupling of beta2-adrenoceptors to multiple G-proteins in the failing human heart. A1 - Harding, SE A1 - Gong, H J1 - Congest Heart Fail Y1 - 2004/07// VL - 10 SN - 1527-5299 N2 - Beta blockers have been shown in clinical trials to improve cardiac function and reduce mortality of heart failure patients. However, these agents require careful titration since they can produce an initial decrease in cardiac output. The authors have recently shown that beta blockers, including some used clinically, can directly depress contraction of myocardium from the failing (but not nonfailing) human heart. This occurs on single ventricular myocytes and is therefore completely independent of any inhibition of endogenous catecholamines. The effect appears to be mediated primarily by the beta2-adrenoceptor (AR) and is dependent on the inhibitory guanine nucleotide binding protein, Gi. Using a transgenic mouse model, as well as adenoviral vectors to overexpress Gi or the human beta2AR in adult myocytes of various species, the authors demonstrate that agents that are blockers for bAR/Gs coupling can be agonists at a beta2AR/Gi-coupled form of the receptor. The negative effect of beta blockers could contribute to the initial cardiodepression that is observed when introducing these agents in heart failure patients. However, in the long term, beta2AR/Gi coupling may enhance the ability of beta blockers to protect and improve the function of the failing heart. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15314476&query_hl=1 ER - TY - JFULL T1 - Nuclear receptor corepressor RIP140 regulates fat accumulation. A1 - Leonardsson, G A1 - Steel, JH A1 - Christian, M A1 - Pocock, V A1 - Milligan, S A1 - Bell, J A1 - So, PW A1 - Medina-Gomez, G A1 - Vidal-Puig, A A1 - White, R A1 - Parker, MG J1 - Proc Natl Acad Sci U S A Y1 - 2004/06/01/ VL - 101 SN - 0027-8424 SP - 8437 EP - 8442 N2 - Nuclear receptors and their coactivators have been shown to function as key regulators of adipose tissue biology. Here we show that a ligand-dependent transcriptional repressor for nuclear receptors plays a crucial role in regulating the balance between energy storage and energy expenditure. Mice devoid of the corepressor protein RIP140 are lean, show resistance to high-fat diet-induced obesity and hepatic steatosis, and have increased oxygen consumption. Although the process of adipogenesis is unaffected, expression of certain lipogenic enzymes is reduced. In contrast, genes involved in energy dissipation and mitochondrial uncoupling, including uncoupling protein 1, are markedly increased. Therefore, the maintenance of energy homeostasis requires the action of a transcriptional repressor in white adipose tissue, and ligand-dependent recruitment of RIP140 to nuclear receptors may provide a therapeutic target in the treatment of obesity and related disorders. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15155905&query_hl=1 ER - TY - JFULL T1 - Control of human articular chondrocyte differentiation by reduced oxygen tension. A1 - Murphy, CL A1 - Polak, JM J1 - J Cell Physiol Y1 - 2004/06// VL - 199 SN - 0021-9541 SP - 451 EP - 459 N2 - Cell number is often a limiting factor in studies of chondrocyte physiology, particularly for human investigations. Chondrocytes can be readily proliferated in monolayer culture, however, differentiated phenotype is soon lost. We therefore endeavored to restore normal phenotype to human chondrocytes after serial passage in monolayer culture by manipulating cell morphology and oxygen tension towards the in vivo state. Third passage cells were encapsulated in alginate and exposed to either 20% or more physiologic 5% oxygen tensions. To assess cell phenotype, gene expression was measured using TaqMan real-time PCR. Encapsulated, primary chondrocytes cultured in 20% oxygen were used as a positive reference. Passaged human chondrocytes were fibroblastic in appearance and had lost normal phenotype as evidenced by a decrease in expression of collagen II, aggrecan, and sox9 genes of 66, 6, and 14 fold, respectively; with concomitant high expression of type I collagen (22 fold increase). A partial regaining of the differentiated phenotype was observed by encapsulation in 20% oxygen; however, even after 4 weeks, collagen II gene expression was not fully restored. Collagen II and aggrecan expression were increased, on average, 3 fold, in 5% oxygen tension compared to 20% cultures. Furthermore, matrix glycosaminoglycan (GAG) levels were significantly increased in reduced oxygen. In fact, after 4 weeks in 5% oxygen, encapsulated third passage cells had collagen II expression fully regained and aggrecan and sox9 levels actually exceeding primary cell levels in 20% oxygen. Our results show that the phenotype of serially passaged human articular chondrocytes is more fully restored by combining encapsulation with culture in more physiological levels of oxygen. Sox9, an essential transcription factor for chondrocyte differentiation is strongly implicated in this process since its expression was upregulated almost 27 fold. These findings have implications for the optimal conditions for the in vitro culture of chondrocytes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15095292&query_hl=1 ER - TY - JFULL T1 - In situ spectral monitoring of mRNA translation in embryonic stem cells during differentiation in vitro. A1 - Notingher, I A1 - Bisson, I A1 - Bishop, AE A1 - Randle, WL A1 - Polak, JM A1 - Hench, LL J1 - Anal Chem Y1 - 2004/06/01/ VL - 76 SN - 0003-2700 SP - 3185 EP - 3193 N2 - Raman microspectroscopy was used to determine biochemical markers during the differentiation of embryonic murine stem cells (mES) in vitro. Such markers are useful to determine the differentiation status of ES cells cultured on biomaterials. Raman spectra of mES cells as undifferentiated, spontaneously differentiated (4 days), and differentiated cells via formation of embryoid bodies (16, 20 days) were analyzed. Unsupervised hierarchical cluster analysis and principal component analysis were used to determine biochemical differences between mES cells in various states of differentiation. The undifferentiated cells were characterized by high scores of the first principal component (PC1, 49% variance). Similarity between the PC1 loading and the Raman spectrum of RNA indicated a high concentration of RNA in mES cells compared to differentiated cells. The ratio between the peak areas of RNA and proteins was used as a measure of mRNA translation. Using the same peak area ratio, it was possible to differentiate even between mES as undifferentiated and in early stages of differentiation (4 days). These findings were correlated with biological studies reporting high levels of nontranslated mRNA during early embryonic development. Therefore, the RNA translation obtained from the Raman spectra can be used as marker of differentiation state of mES cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15167800&query_hl=1 ER - TY - JFULL T1 - A histological study of the hph-1 mouse mutant: an animal model of phenylketonuria and infantile hypertrophic pyloric stenosis A1 - Abel, RM A1 - Dore, CJ A1 - Bishop, AE A1 - Facer, P A1 - Polak, JM A1 - Spitz, L J1 - ANAT HISTOL EMBRYOL Y1 - 2004/06// VL - 33 SN - 0340-2096 SP - 125 EP - 130 N2 - Aim: To quantify the chronological sequence of changes in the morphology and immunoreactivity for neurotransmitters in the pylorus of an animal model of infantile hypertrophic pyloric stenosis and phenylketonuria. Method: Thirty specimens of pylorus from hph-1 mice and age/sex matched controls (age range: 10-180 days) were examined using conventional histology and immunohistochemistry for a variety of antigens: protein gene product 9.5, a pan neuronal marker; vasoactive intestinal polypeptide; nitric oxide synthase two antigens coalesced to the same inhibitory neurons in humans; substance P, a potent excitatory neurotransmitter; and calcitonin gene related peptide, a neurotransmitter implicated in the somatic afferent innervation of the stomach. The changes in the morphology of the muscle layers were quantified and statistically analysed for each age group (10, 20, 40, 90 and 180 days). Results: Between 10 and 90 days of age, all muscle layers of the hph-1 mice were hypertrophied, for example, 10 days, hph-1 longitudinal muscle mean diameter = 3.4, control = 1.8; hph-1 circular muscle width = 11.5, control = 4.7. The hph-1 mice were significantly smaller during this period (40 days, hph-1 weight = 10 g, control = 25 g). There was no change in the pattern of expression of the antigens examined within the hph-1 mice compared with the controls. Conclusion: Hph-1 mice develop a transient smooth muscle hypertrophy of the pylorus attended by gastric distension and failure to gain weight. These changes resolve as the pyloric muscle hypertrophy resolves. ER - TY - JFULL T1 - Clinical recovery from end-stage heart failure using left-ventricular assist device and pharmacological therapy correlates with increased sarcoplasmic reticulum calcium content but not with regression of cellular hypertrophy. A1 - Terracciano, CM A1 - Hardy, J A1 - Birks, EJ A1 - Khaghani, A A1 - Banner, NR A1 - Yacoub, MH J1 - Circulation Y1 - 2004/05/18/ VL - 109 SN - 1524-4539 SP - 2263 EP - 2265 N2 - BACKGROUND: Left ventricular assist device (LVAD) treatment is known to lead to structural and functional cellular modifications in the heart. The relevance of these changes for clinical recovery is unknown. METHODS AND RESULTS: We compared properties of cardiomyocytes obtained from tissue taken at explantation of the LVAD in patients with clinical recovery with those obtained from hearts of patients who did not show clinical recovery, thus requiring transplantation. Compared with myocytes taken at implantation, both the recovery and nonrecovery groups showed approximately 50% reduction in cell capacitance, an index of cell size. However, action potential duration shortened, L-type Ca2+ current fast inactivation was more rapid, and sarcoplasmic reticulum Ca2+ content was increased in the recovery compared with the nonrecovery group. CONCLUSIONS: These results show that specific changes in excitation-contraction coupling, and not regression of cellular hypertrophy, are specifically associated with clinical recovery after LVAD and further identify sarcoplasmic reticulum Ca2+ handling as a key functional determinant in patients with heart failure. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15136495&query_hl=1 ER - TY - JFULL T1 - Functional characteristics of the cardiac ryanodine receptor in an experimental model of heart failure A1 - Scoote, M A1 - Harding, SE A1 - MacLeod, KT A1 - Kingsbury, MP A1 - Sheridan, DJ A1 - Williams, AJ J1 - J MOL CELL CARDIOL Y1 - 2004/05// VL - 36 SN - 0022-2828 SP - 757 EP - 758 ER - TY - JFULL T1 - Modulation of contractility by gene therapy A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2004/05// VL - 36 SN - 0022-2828 SP - 731 EP - 731 ER - TY - JFULL T1 - Characterisation of beta-adrenoceptor responses in mouse embryonic stem cell-derived cardiomyocytes A1 - Ali, NN A1 - Xu, X A1 - Martins, M A1 - Poole-Wilson, PA A1 - Harding, SE A1 - Fuller, SJ J1 - J MOL CELL CARDIOL Y1 - 2004/05// VL - 36 SN - 0022-2828 SP - 731 EP - 732 ER - TY - JFULL T1 - Osteogenic differentiation of mouse embryonic stem cells: differential gene expression analysis by cDNA microarray and purification of osteoblasts by cadherin-11 magnetically activated cell sorting. A1 - Bourne, S A1 - Polak, JM A1 - Hughes, SP A1 - Buttery, LD J1 - Tissue Eng Y1 - 2004/05// VL - 10 SN - 1076-3279 SP - 796 EP - 806 N2 - We have previously shown osteogenic differentiation of mouse embryonic stem (ES) cells and temporal enrichment with osteoblastic cells, by stimulation with serum-containing culture medium supplemented with beta-glycerophosphate, ascorbate, and dexamethasone. In our present study we have used similar culture conditions to further investigate osteogenic differentiation of mouse ES cells. Using reverse transcription-polymerase chain reaction (RT-PCR) we demonstrated the expression of genes associated with osteoblast differentiation including the bone matrix protein osteocalcin and the transcription factor Cbfa-1/runx2. Furthermore, results of cDNA microarray analysis, and subsequent RT-PCR analysis of differentiating ES cells after exposure to osteogenic stimuli, revealed a combination of upregulation of genes involved in osteoblast differentiation including osteopontin, HSP-47, and IGF-II coupled with downregulation of genes involved in differentiation of other phenotypes such as the neuroectoderm factor Stra-13. Finally, we have applied magnetically activated cell-sorting methods to ES cell cultures treated with osteogenic stimuli and, using an antibody to cadherin-11, have purified a subpopulation of cells with osteoblastic characteristics. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15265297&query_hl=1 ER - TY - JFULL T1 - Altered cardiac ryanodine receptor: dihydropyridine receptor stoichiometry in an experimental model of heart failure A1 - Scoote, M A1 - Harding, SE A1 - MacLeod, KT A1 - Kingsbury, MP A1 - Sheridan, DJ A1 - Williams, AJ J1 - J MOL CELL CARDIOL Y1 - 2004/05// VL - 36 SN - 0022-2828 SP - 732 EP - 732 ER - TY - JFULL T1 - Phosphatidylinositol-3 kinase inhibitors reproduce the selective antiproliferative effects of imatinib on chronic myeloid leukaemia progenitor cells. A1 - Marley, SB A1 - Lewis, JL A1 - Schneider, H A1 - Rudd, CE A1 - Gordon, MY J1 - Br J Haematol Y1 - 2004/05// VL - 125 SN - 0007-1048 SP - 500 EP - 511 N2 - We investigated the role of the phosphatidylinositol-3 kinase (PI-3K) pathway in regulating the proliferation of primary chronic myeloid leukaemia (CML) progenitor cells by using imatinib to inhibit the activity of p210(Bcr-Abl). The effect of imatinib on the expression of PI-3K pathway proteins was investigated by kinase assays and Western blotting; PI-3K was inhibited by wortmannin or LY294002, Jak2 by AG490 and farnesylation by FTI II; progenitor cell proliferation (self-renewal) was measured by growing myeloid colonies in vitro, then replating them to observe secondary colony formation. Suppression of p210(Bcr-Abl) with imatinib indirectly suppressed the activity of PI-3K and its downstream targets (Erk, Akt and p70S6 kinase), thereby implicating the PI-3K pathway in p210(Bcr-Abl)-mediated signalling in primary CML progenitor cells. The PI-3K inhibitors, wortmannin and LY294002 reproduced the differential effects of imatinib on normal and CML progenitor cell proliferation in vitro by increasing normal cell (P = 0.001) and reducing CML cell proliferation (P = 0.0003). This differential effect was attributable to dysregulated signalling by granulocyte colony-stimulating factor in CML. The responses of individual patient's cells to wortmannin correlated with their responses to imatinib (P = 0.004) but not their responses to AG490 (Jak2 kinase inhibitor) or FTI II (farnesyltransferase inhibitor). Individual responses to wortmannin also correlated with responses to interferon alpha (IFNalpha) (P = 0.016). Imatinib-resistant K562 cells were sensitive to LY294002. Inhibition of the PI-3K pathway may be common to imatinib and IFNalpha and reflect dysregulated cytokine signalling. As imatinib-resistant cells remained sensitive to wortmannin and LY294002, targeting the PI-3K pathway may provide an alternative therapy for imatinib-resistant patients. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15142121&query_hl=1 ER - TY - JFULL T1 - A membrane-mimetic barrier for islet encapsulation. A1 - Cui, W A1 - Barr, G A1 - Faucher, KM A1 - Sun, XL A1 - Safley, SA A1 - Weber, CJ A1 - Chaikof, EL J1 - Transplant Proc Y1 - 2004/05// VL - 36 SN - 0041-1345 SP - 1206 EP - 1208 N2 - BACKGROUND: Enhanced control of both transport properties and surface physiochemical characteristics will be important steps in the development of an effective immunoisolation barrier critical to the success of pancreatic islet cell transplantation. We hypothesize that the cell membrane establishes an important paradigm for the design of a biomimetic immunoisolation barrier with improved performance characteristics because of its capacity to control interfacial mass transport, as well as its ability to act as a template for more complex structures with other immunoregulatory macromolecules. METHODS: Islets were isolated from Wistar rats using collagenase digestion and a discontinuous Ficoll-Histopaque gradient and subsequently encapsulated in 2% alginate. After coating with a polyelectrolyte multilayer of polylysine and alginate, a polymeric membrane-mimetic coating was applied to the capsule surface. Individual islet viability was evaluated at each stage of the encapsulation procedure by use of a two-color live/dead cell assay. Preservation of islet function was determined by transplanting 1000 encapsulated islets into the peritoneal cavity of streptozotocin-induced diabetic nonobese diabetic NOD/Scid mice. RESULTS: At the end of the coating procedure, the proportion of viable cells within each islet was >50% in 88% of encapsulated rat islets and >75% in over half of the encapsulated cohort. Nonfasting blood glucose levels normalized within 24 hours after transplantation (n = 8). Normoglycemia has been maintained in all mice with the longest time course being 73 days thus far. CONCLUSIONS: We have demonstrated that microencapsulated islets coated with a membrane-mimetic thin film can be generated with high viability in vitro and persistent function in vivo. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15194418&query_hl=1 ER - TY - JFULL T1 - Photodynamic therapy significantly improves survival outcomes in people with non-resectable cholangiocarcinoma. A1 - Khan, SA A1 - Sharif, AW A1 - Taylor-Robinson, SD J1 - Cancer Treat Rev Y1 - 2004/05// VL - 30 SN - 0305-7372 SP - 315 EP - 318 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15059655&query_hl=1 ER - TY - JFULL T1 - Characterization of four autonomous repression domains in the corepressor receptor interacting protein 140. A1 - Christian, M A1 - Tullet, JM A1 - Parker, MG J1 - J Biol Chem Y1 - 2004/04/09/ VL - 279 SN - 0021-9258 SP - 15645 EP - 15651 N2 - Receptor interacting protein (RIP) 140 is a corepressor that can be recruited to nuclear receptors by means of LXXLL motifs. We have characterized four distinct autonomous repression domains in RIP140, termed RD1-4, that are highly conserved in mammals and birds. RD1 at the N terminus represses transcription in the presence of trichostatin A, suggesting that it functions by a histone deacetylase (HDAC)-independent mechanism. The repressive activity of RD2 is dependent upon carboxyl-terminal binding protein recruitment to two specific binding sites. Use of specific inhibitors indicates that RD2, RD3, and RD4 are capable of functioning by HDAC-dependent and HDAC-independent mechanisms, depending upon cell type. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14736873&query_hl=1 ER - TY - JFULL T1 - Beta-blockers, myocardial ischaemia and collateral circulation. A1 - Sato, M A1 - Harding, SE A1 - Poole-Wilson, PA J1 - Eur Heart J Y1 - 2004/04// VL - 25 SN - 0195-668X SP - 537 EP - 539 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15120048&query_hl=1 ER - TY - JFULL T1 - VERITAS: the Very Energetic Radiation Imaging Telescope Array System A1 - Krennrich, F A1 - Bond, IH A1 - Boyle, PJ A1 - Bradbury, SM A1 - Buckley, JH A1 - Carter-Lewis, D A1 - Celik, O A1 - Cui, W A1 - Daniel, M A1 - D'Vali, M A1 - Perez, IDC A1 - Duke, C A1 - Falcone, A A1 - Fegan, DJ A1 - Fegan, SJ A1 - Finley, JP A1 - Fortson, LF A1 - Gaidos, J A1 - Gammell, S A1 - Gibbs, K A1 - Gillanders, GH A1 - Grube, J A1 - Hall, J A1 - Hall, TA A1 - Hanna, D A1 - Hillas, AM A1 - Holder, J A1 - Horan, D A1 - Jarvis, A A1 - Kenny, GE A1 - Kertzman, M A1 - Kieda, D A1 - Kildea, J A1 - Knapp, J A1 - Kosack, K A1 - Krawczynski, H A1 - Lang, MJ A1 - LeBohec, S A1 - Linton, E A1 - Lloyd-Evans, J A1 - Milovanovic, A A1 - Moriarty, P A1 - Muller, D A1 - Nagai, T A1 - Nolan, S A1 - Ong, RA A1 - Pallassini, R A1 - Petry, D A1 - Power-Mooney, B A1 - Quinn, J A1 - Quinn, M A1 - Ragan, K A1 - Rebillot, P A1 - Reynolds, PT A1 - Rose, HJ A1 - Schroedter, M A1 - Sembroski, G A1 - Swordy, SP A1 - Syson, A A1 - Vassiliev, VV A1 - Walker, G A1 - Wakely, SP A1 - Weekes, TC A1 - Zweerink, J J1 - NEW ASTRON REV Y1 - 2004/04// VL - 48 SN - 1387-6473 SP - 345 EP - 349 N2 - The Very Energetic Radiation Imaging Telescope Array System (VERITAS) is the major next generation imaging atmospheric Cherenkov gamma-ray telescope in the western hemisphere and will be located in southern Arizona nearby Kitt Peak National Observatory. The VERITAS observatory will provide unprecedented sensitivity to photon energies between 50 GeV and 50 TeV. The first stage is an array of four telescopes to be fully operational in early 2006, with an expansion to seven telescopes envisioned for 2008. The construction of a prototype telescope is underway, for which first light is expected in Fall 2003. The technical concept is outlined and a progress report is given. (C) 2004 Published by Elsevier B.V. ER - TY - JFULL T1 - Overexpression of beta(1)-adrenoceptors in adult rat ventricular myocytes enhances CGP 12177A cardiostimulation: implications for 'putative' beta(4)-adrenoceptor pharmacology A1 - Lewis, CJ A1 - Gong, HB A1 - Brown, MJ A1 - Harding, SE J1 - BRIT J PHARMACOL Y1 - 2004/03// VL - 141 SN - 0007-1188 SP - 813 EP - 824 N2 - 1 CGP 12177A mediates cardiostimulation by activation of the 'putative' beta(4)-adrenoceptor; however, it has recently been reported that disruption of the beta(1)-adrenoceptor gene abolishes this effect. We have adenovirally overexpressed beta(1)-adrenoceptors in isolated, cultured adult rat ventricular cardiomyocytes and observed the inotropic potency of isoprenaline and CGP 12177A ( in the presence of 1 muM propranolol).2 Isoprenaline was a full inotropic agonist at rat ventricular myocytes (pD(2) 7.69 +/- 0.12). CGP 12177A was a nonconventional partial agonist (pD(2) 6.34 +/- 0.09), increasing inotropy and lusitropy, with an intrinsic activity of 0.34 and antagonised by bupranolol.3 beta(1)-adrenoceptor overexpression enhanced the inotropic potency of isoprenaline by 11.7-fold (pD(2) 8.76 +/- 0.14) and CGP 12177A by 5.9-fold (7.11 +/- 0.10), respectively. Green fluorescent protein (GFP) overexpression did not alter the potency of isoprenaline or CGP 12177A (pD(2) 7.41 +/- 0.24 and pD(2) 6.60 +/- 0.50, respectively).4 The cardiostimulant effects of CGP 12177A were enhanced by IBMX ( phosphodiesterase inhibitor) and decreased by Rp-cAMPS ( cAMP antagonist). CGP 12177A also increased cAMP levels. CGP 12177A but not isoprenaline initiated arrhythmias at lower concentrations following beta(1)-adrenoceptor overexpression.5 I-125-Cyanopindolol saturation binding in Adv.beta(1) myocytes demonstrated similar to18-fold increase in beta(1)-adrenoceptors. H-3-CGP 12177A saturation binding, in the presence of propranolol, increased similar to5-fold following overexpression of beta(1)-adrenoceptors.6 This study demonstrates enhanced cardiostimulation by CGP 12177A ( in the presence of propranolol) in rat ventricular myocytes overexpressing beta(1)-adrenoceptors, mediated by a Gs/cAMP signalling pathway. 'Putative' beta(4)-adrenoceptor pharmacology appears to be mediated by activation of a novel affinity state of the beta(1)-adrenoceptor. ER - TY - JFULL T1 - The stem cell in orthopaedic surgery. A1 - Vats, A A1 - Tolley, NS A1 - Buttery, LD A1 - Polak, JM J1 - J Bone Joint Surg Br Y1 - 2004/03// VL - 86 SN - 0301-620X SP - 159 EP - 164 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15046426&query_hl=1 ER - TY - JFULL T1 - Embryonic stem cells. A1 - Rippon, HJ A1 - Bishop, AE J1 - Cell Prolif Y1 - 2004/02// VL - 37 SN - 0960-7722 SP - 23 EP - 34 N2 - Embryonic stem cells have huge potential in the field of tissue engineering and regenerative medicine as they hold the capacity to produce every type of cell and tissue in the body. In theory, the treatment of human disease could be revolutionized by the ability to generate any cell, tissue, or even organ, 'on demand' in the laboratory. This work reviews the history of murine and human ES cell lines, including practical and ethical aspects of ES cell isolation from pre-implantation embryos, maintenance of undifferentiated ES cell lines in the cell culture environment, and differentiation of ES cells in vitro and in vivo into mature somatic cell types. Finally, we discuss advances towards the clinical application of ES cell technology, and some of the obstacles which must be overcome before large scale clinical trials can be considered. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14871235&query_hl=1 ER - TY - JFULL T1 - Gene transfer in cardiac myocytes. A1 - Chaudhri, BB A1 - del Monte, F A1 - Harding, SE A1 - Hajjar, RJ J1 - Surg Clin North Am Y1 - 2004/02// VL - 84 SN - 0039-6109 N2 - Congestive heart failure (CHF) represents an enormous clinical problem and remains a leading cause of death despite advances in treatment. New treatments significantly impact mortality and disease course; they do not cure the underlying pathology. Gene transfer, the ability to genetically reprogram the heart in relevant cardiovascular disease models, allows testing the role of specific molecular pathways in disease pathogenesis. Potential therapeutic intervention targets can be then identified and approached with the full spectrum of therapeutic options, including traditional pharmacology, targeted synthesis of small molecule agonists or antagonists, biological agents (cells, antibodies, genetic material), or gene-based therapy. Lessons gleaned from gene transfer experiments on local modulation of cardiac genetic programs will guide attempts to transform early investigations into established therapy. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15053187&query_hl=1 ER - TY - JFULL T1 - Pulmonary epithelial stem cells. A1 - Bishop, AE J1 - Cell Prolif Y1 - 2004/02// VL - 37 SN - 0960-7722 SP - 89 EP - 96 N2 - Classically, the stem/progenitor cells of the pulmonary epithelium are considered to be the basal and mucous cells of the proximal airways, Clara cells in the bronchioles and type II pneumocytes in the alveoli. Recent data suggest that there is a variant of Clara cells, lying in pulmonary neuroendocrine bodies, that meets several stem cell criteria and that type II pneumocytes exist in at least two populations, one of which is more resistant to injury. However, a complete revision of our understanding of pulmonary stem cell biology is underway as a result of the discovery of pulmonary epithelium derived from blood-borne cells. In addition, the existence in the lung of a 'universal' pluripotent cell has long been speculated upon and now some initial evidence has emerged with the identification of a spore-like cell that can differentiate in vitro to bronchiolar tissue. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14871239&query_hl=1 ER - TY - JFULL T1 - Role of interleukin 6 in myocardial dysfunction of meningococcal septic shock. A1 - Pathan, N A1 - Hemingway, CA A1 - Alizadeh, AA A1 - Stephens, AC A1 - Boldrick, JC A1 - Oragui, EE A1 - McCabe, C A1 - Welch, SB A1 - Whitney, A A1 - O'Gara, P A1 - Nadel, S A1 - Relman, DA A1 - Harding, SE A1 - Levin, M J1 - Lancet Y1 - 2004/01/17/ VL - 363 SN - 1474-547X SP - 203 EP - 209 N2 - BACKGROUND: Myocardial failure has a central role in the complex pathophysiology of septic shock and contributes to organ failure and death. During the sepsis-induced inflammatory process, specific factors are released that depress myocardial contractile function. We aimed to identify these mediators of myocardial depression in meningococcal septic shock. METHODS: We combined gene-expression profiling with protein and cellular methods to identify a serum factor causing cardiac dysfunction in meningococcal septic shock. We identified genes that were significantly upregulated in blood after exposure to meningococci. We then selected for further analysis those genes whose protein products had properties of a myocardial depressant factor--specifically a 12-25 kDa heat-stable protein that is released into serum shortly after onset of meningococcal infection. FINDINGS: We identified 174 significantly upregulated genes in meningococcus-infected blood: six encoded proteins that were of the predicted size and had characteristics of a myocardial depressant factor. Of these, interleukin 6 caused significant myocardial depression in vitro. Removal of interleukin 6 from serum samples of patients with meningococcaemia and from supernatants of inflammatory cells stimulated by meningococci in vitro abolished the negative inotropic activity. Furthermore, concentrations in serum of interleukin 6 strongly predicted degree of myocardial dysfunction and severity of disease in children with meningococcal septic shock. INTERPRETATION: Interleukin 6 is a mediator of myocardial depression in meningococcal disease. This cytokine and its downstream mediators could be a target for future treatment strategies. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14738793&query_hl=1 ER - TY - JFULL T1 - The hemopoietic cytokine Flt3 ligand increases the resistance of mice to a lethal burn wound infection A1 - Toliver-Kinsky, TE A1 - Cui, W A1 - Herndon, DN A1 - Sherwood, ER J1 - SHOCK Y1 - 2004/01// VL - 21 SN - 1073-2322 SP - 18 EP - 18 ER - TY - JFULL T1 - In-situ monitoring of cell death using Raman microspectroscopy A1 - Verrier S A1 - Notingher I A1 - Polak JM A1 - Hench LL J1 - Biopolymers Y1 - 2004/// VL - 74 SP - 157 EP - 162 ER - TY - JFULL T1 - Loss of beta-adrenoceptor response in myocytes overexpressing the Na+/Ca(2+)-exchanger. A1 - Sato, M A1 - Gong, H A1 - Terracciano, CM A1 - Ranu, H A1 - Harding, SE J1 - J Mol Cell Cardiol Y1 - 2004/01// VL - 36 SN - 0022-2828 SP - 43 EP - 48 N2 - Increased Na+/Ca(2+)-exchanger (NCX) and altered beta-adrenoceptor (betaAR) responses are observed in failing human heart. To determine the possible interaction between these changes, we investigated the effect of NCX overexpression on responses to isoproterenol in adult rat ventricular myocytes. Responses to isoproterenol were largely mediated through the beta1AR in control myocytes. Adenovirally-mediated overexpression of NCX, at levels, which did not alter basal contraction of myocytes, markedly depressed the isoproterenol concentration-response curve. Responses to isoproterenol could be restored to normal by beta2AR blockade, suggesting a beta2AR-mediated inhibition of beta1AR signalling. Pertussis toxin normalised isoproterenol responses in NCX cells, indicating that beta2AR effects were mediated by Gi. Negative-inotropic effects of high concentrations of ICI 118,551, previously shown to be due to beta2AR-Gi coupling, were increased in NCX cells. We conclude that NCX upregulation can markedly alter the consequences of betaAR stimulation and that this may contribute to the alterations in betaAR response seen in failing human heart. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14734046&query_hl=1 ER - TY - JFULL T1 - Bioactive glasses for in situ tissue regeneration. A1 - Hench, LL A1 - Xynos, ID A1 - Polak, JM J1 - J Biomater Sci Polym Ed Y1 - 2004/// VL - 15 SN - 0920-5063 SP - 543 EP - 562 N2 - Historically the function of biomaterials has been to replace diseased or damaged tissues. Recent findings show that controlled release of the ionic dissolution products of bioactive glasses results in regeneration of tissues. The mechanism for in situ tissue regeneration involves upregulation of seven families of genes that control the osteoblast cell cycle, mitosis and differentiation. In the presence of critical concentrations of Si and Ca ions, within 48 h osteoblasts that are capable of differentiating into a mature osteocyte phenotype begin to proliferate and regenerate new bone. Osteoblasts that are not in the correct phase of the cell cycle and unable to proceed towards differentiation are switched into apoptosis by the ionic dissolution products. A controlled release of soluble Ca and Si from bioactive glass--resorbable polymer composites leads to vascularised soft tissue regeneration. Gene activation by controlled ion release provides the conceptual basis for molecular design of a third generation of biomaterials optimised for in situ tissue regeneration. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15212333&query_hl=1 ER - TY - JFULL T1 - Initial observations on the effect of medium composition on the differentiation of murine embryonic stem cells to alveolar type II cells. A1 - Rippon, HJ A1 - Ali, NN A1 - Polak, JM A1 - Bishop, AE J1 - Cloning Stem Cells Y1 - 2004/// VL - 6 SN - 1536-2302 SP - 49 EP - 56 N2 - The pluripotency and high proliferative index of embryonic stem (ES) cells make them a good potential source of cells for tissue engineering purposes. We have shown that ES cells can be induced to differentiate in vitro into pulmonary epithelial cells (type II pneumocytes) using a serum-free medium designed for the maintenance of mature distal lung epithelial cells in culture (SAGM). However, the resulting cell cultures were heterogeneous. Our aim in this study was to attempt to increase pneumocyte yield and differentiation state by determining which medium components enhance the differentiation of pneumocytes and modifying the medium accordingly. Quantitative RT-PCR was used to measure changes in the expression of a type II pneumocyte-specific gene, surfactant protein C (SPC), in response to alterations in the cell culture medium. Results suggested that most individual SAGM growth factors were inhibitory for type II pneumocyte differentiation, with the largest increases in SPC expression (approximately threefold) being observed upon removal of retinoic acid and triiodothryonine. However, large standard deviations occurred between replicates, illustrating the highly variable nature of ES cell differentiation. Nevertheless, these observations represent an initial step towards achieving directed differentiation of pneumocytes from stem cells that could lead to their purification for tissue engineering purposes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15268776&query_hl=1 ER - TY - JFULL T1 - Profibrotic effects of endothelin-1 via the ETA receptor in cultured human cardiac fibroblasts. A1 - Hafizi, S A1 - Wharton, J A1 - Chester, AH A1 - Yacoub, MH J1 - Cell Physiol Biochem Y1 - 2004/// VL - 14 SN - 1015-8987 SP - 285 EP - 292 N2 - BACKGROUND/AIMS: Endothelin-1 (ET-1) has been implicated in pathologic remodelling and tissue repair processes in the heart. We investigated the effects of ET-1 on growth and collagen synthesis responses in cardiac fibroblasts isolated from human hearts. We also studied the receptor subtype(s) mediating such responses and the factors regulating their expression. METHODS: Fibroblasts were isolated from cardiac transplant recipient hearts and characterised by immunocytochemistry. Serum-starved cells were exposed to ET-1 and incorporation of [3H]proline and thymidine were measured as indexes of collagen and DNA synthesis respectively. Blocking experiments utilised the selective ETA receptor antagonist BQ123 and the ETB antagonist BQ788. RESULTS: ET-1 elicited a potent collagen synthesis response in cardiac fibroblasts, with a maximum 29+/-5% increase that was abolished by BQ123. Cardiac fibroblasts responded to ET-1 with a concentration-dependent decrease in DNA synthesis rate. The effects of ET-1 were similar to those of TGF-beta. Radioligand binding studies revealed the presence of high-affinity ET-1 binding sites on these cells, which were upregulated by treatment with the growth factors PDGF and EGF but downregulated by TGF-beta. CONCLUSIONS: These results therefore implicate ET-1 as a trophic agent in the human heart with the ability to influence the development of cardiac fibrosis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15319532&query_hl=1 ER - TY - JFULL T1 - Grid gateway: Message-passing between separated cluster interconnects A1 - Cui, W A1 - Ma, J A1 - Huo, ZG J1 - LECT NOTES COMPUT SC Y1 - 2004/// VL - 3032 SN - 0302-9743 SP - 724 EP - 731 N2 - Geographically distributed computing requires high-performance clusters to be integrated to solve problems in computational Grid. Because cluster interconnect is isolated, its low-level communication protocol doesn't exchange messages with others directly. This paper presents a plug-in, Grid Gateway, which enables separated low-level communication protocols to communicate with each other. Grid Gateway can be used in many topologies of inter-cluster network. It has some dynamic features, such as support for multi-gateway mechanism to enhance communication performance. Grid Gateway allows low-level communication protocol to involve in the high-performance Grid computing. Thus it is expected to support the implementation of Grid-enabled tools over it, such as Grid-enabled MPI. This paper describes its architecture and implementation, and presents some design issues. ER - TY - JFULL T1 - Opposing effects of PI3 kinase pathway activation on human myeloid and erythroid progenitor cell proliferation and differentiation in vitro. A1 - Lewis, JL A1 - Marley, SB A1 - Ojo, M A1 - Gordon, MY J1 - Exp Hematol Y1 - 2004/01// VL - 32 SN - 0301-472X SP - 36 EP - 44 N2 - OBJECTIVE: To investigate 1) the effects of lineage-specific cytokines (G-CSF and EPO) combined with ligands for different classes of cytokine receptors (common beta chain, gp130, and tyrosine kinase) on proliferation by human myeloid and erythroid progenitor cells; and 2) the signal transduction pathways associated with combinatorial cytokine actions. PATIENTS AND METHODS: CFU-GM and BFU-E were cloned in vitro. Secondary colony formation by replated CFU-GM and subcolony formation by BFU-E provided measures of progenitor cell proliferation. Studies were performed in the presence of cytokine combinations with and without signal transduction inhibitors. RESULTS: Proliferation by CFU-GM and BFU-E was enhanced synergistically when common beta chain receptor cytokines (IL-3 or GM-CSF) were combined with G-CSF or EPO, but not with gp130 receptor cytokines (LIF or IL-6) or tyrosine kinase receptor cytokines (SCF, HGF, Flt-3 ligand, or PDGF). Delayed addition studies with G-CSF+IL-3 and EPO+IL-3 demonstrated that synergy required the presence of both cytokines from the initiation of the culture. The Jak2-specific inhibitor, AG490, abrogated the effect of combining IL-3 with EPO but had no effect on the enhanced CFU-GM proliferation stimulated by IL-3+G-CSF. The PI3 kinase inhibitors LY294002 and wortmannin substituted for G-CSF in combination with IL-3 since proliferation in the presence of LY294002/wortmannin+IL-3 was enhanced to the same extent as in the presence of G-CSF+IL-3. In contrast, LY294002 and wortmannin inhibited proliferation in the presence of EPO and in the presence of EPO+IL-3. CONCLUSION: 1) IL-3 may activate different signal transduction pathways when combined with G-CSF and when combined with EPO; 2) different signal transducing intermediates regulate erythroid and myeloid progenitor cell proliferation; and 3) inhibition of the PI3 kinase pathway suppresses myeloid progenitor cell differentiation and thereby increases proliferation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14725899&query_hl=1 ER - TY - JFULL T1 - Binary CaO-SiO(2) gel-glasses for biomedical applications. A1 - Saravanapavan, P A1 - Jones, JR A1 - Verrier, S A1 - Beilby, R A1 - Shirtliff, VJ A1 - Hench, LL A1 - Polak, JM J1 - Biomed Mater Eng Y1 - 2004/// VL - 14 SN - 0959-2989 SP - 467 EP - 486 N2 - Bioactive materials are routinely used in dental and orthopaedic applications. The concept was first introduced in 1971, with the discovery of 45S5 Bioglass, which is known to develop an interfacial bond between the implant and the host tissue. This glass is composed of SiO(2), CaO, P(2)O(5) and Na(2)O. Since then numerous glasses and glass ceramics with similar compositions have been extensively studied for clinical applications. Until 1990 it was accepted that P(2)O(5) and Na(2)O were necessary components for the glass composition to be bioactive. However, calcium silicate glasses with high SiO(2) content are impossible to produce using the traditional melt-quench method. This is due to the liquid-liquid immiscibility region that is present between 0.02 and 0.3 mole fraction of CaO and in terms of bioactivity, high CaO compositions were inferior to those quaternary bioactive glass compositions already in existence. In the last few years several studies have been reported regarding the production of CaO-SiO(2) glasses via the sol-gel processing technique. This report summarises the findings of the past and the present and also outlines potential of these calcium silicate gel-glasses in the field of biomaterials. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15472395&query_hl=1 ER - TY - JFULL T1 - In-situ spectroscopic study of nucleic acids in differentiating embryonic stem cells. A1 - Notingher I A1 - Bisson I A1 - Polak JM A1 - Hench LL J1 - Vib.Spectrosc. Y1 - 2004/// VL - 35 SP - 199 EP - 203 ER - TY - JFULL T1 - The application of magnetic resonance imaging and spectroscopy to gene therapy. A1 - Bhakoo, KK A1 - Bell, JD A1 - Cox, IJ A1 - Taylor-Robinson, SD J1 - Methods Enzymol Y1 - 2004/// VL - 386 SN - 0076-6879 SP - 303 EP - 313 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15120258&query_hl=1 ER - TY - JFULL T1 - The thyroid hormone receptor-associated protein TRAP220 is required at distinct embryonic stages in placental, cardiac, and hepatic development. A1 - Landles, C A1 - Chalk, S A1 - Steel, JH A1 - Rosewell, I A1 - Spencer-Dene, B A1 - Lalani, el-N A1 - Parker, MG J1 - Mol Endocrinol Y1 - 2003/12// VL - 17 SN - 0888-8809 SP - 2418 EP - 2435 N2 - Recent work indicates that thyroid hormone receptor-associated protein 220 (TRAP220), a subunit of the multiprotein TRAP coactivator complex, is essential for embryonic survival. We have generated TRAP220 conditional null mice that are hypomorphic and express the gene at reduced levels. In contrast to TRAP220 null mice, which die at embryonic d 11.5 (E11.5), hypomorphic mice survive until E13.5. The reduced expression in hypomorphs results in hepatic necrosis, defects in hematopoiesis, and hypoplasia of the ventricular myocardium, similar to that observed in TRAP220 null embryos at an earlier stage. The embryonic lethality of null embryos at E11.5 is due to placental insufficiency. Tetraploid aggregation assays partially rescues embryonic development until E13.5, when embryonic loss occurs due to hepatic necrosis coupled with poor myocardial development as observed in hypomorphs. These findings demonstrate that, for normal placental function, there is an absolute requirement for TRAP220 in extraembryonic tissues at E11.5, with an additional requirement in embryonic tissues for hepatic and cardiovascular development thereafter. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14500757&query_hl=1 ER - TY - JFULL T1 - Interaction between NCX, SERCA2a and beta AR changes in heart failure investigated by adenoviral gene transfer A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2003/11// VL - 35 SN - 0022-2828 SP - A13 EP - A13 ER - TY - JFULL T1 - beta-blockers as activators of Gi-coupled pathways in the failing human heart A1 - Harding, SE J1 - J MOL CELL CARDIOL Y1 - 2003/11// VL - 35 SN - 0022-2828 SP - A43 EP - A43 ER - TY - JFULL T1 - Maintaining transcriptional states through DNA replication. A1 - Azuara, V A1 - Fisher, AG J1 - Cell Cycle Y1 - 2003/11// VL - 2 SN - 1538-4101 SP - 521 EP - 524 N2 - The timing of DNA replication has been implicated in gene regulation based on observations that actively transcribed genes generally replicate earlier in S-phase than their inactive counterparts. However, we recently showed that differentiation-induced gene silencing in lymphocytes does not generally result in silenced genes switching from early to late replication, but instead alters the onset of separation of newly synthesised sister-chromatids. Our findings may provide novel insights into the mechanisms that allow inactive chromatin structure to be propagated through DNA replication. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14504465&query_hl=1 ER - TY - JFULL T1 - Preparation and characterization of molecular weight standards of low polydispersity from oat and barley (1 -> 3)(1 -> 4)-beta-D-glucan A1 - Wang, Q A1 - Wood, PJ A1 - Huang, X A1 - Cui, W J1 - FOOD HYDROCOLLOID Y1 - 2003/11// VL - 17 SN - 0268-005X SP - 845 EP - 853 N2 - Purified (1 --> 3)(1 --> 4)-beta-D-glucans (beta-glucans) from oat and barley with broad molecular weight (MW) distribution were separated into seven fractions using gradient precipitation with ammonium sulfate (NH4)(2)SO4. The MW of each fraction decreased consecutively with the concentration of (NH4)(2)SO4 at which it was precipitated. The MW distribution of each fraction was much narrower compared to the parent sample and is comparable to commercially available pullulan MW standards. To determine whether the fractionation process was separating sub-fractions of different structure, the original beta-glucan sample and each fraction were hydrolyzed by a (1 --> 3)(1 --> 4)-D-beta-glucan-4-glucanohydrolase (lichenase, E.C.3.2.1.73) and the liberated oligosaccharides were analyzed by high performance anion exchange chromatography. The analysis revealed no differences in oligosaccharide pattern (DP 2-9) derived from each fraction and the parent sample. In particular, the tri/tetra. oligosaccharide ratio remained constant for all fractions, indicating no fractionation based on structural features had taken place. The effect of starting beta-glucan concentration on the fractionation process was studied. The results showed that it was possible to achieve good separation at overlapping parameter c[eta] lower than similar to3.5. Further increase in starting beta-glucan concentration hindered clear separation of the fractions. Temperature also affected the fractionation efficiency. The higher the temperature, the lower the amount of (NH4)(2)SO4 that was necessary to precipitate the samples of same MW. A Mark Houwink relationship was derived from the measured MW and intrinsic viscosity for fractions from oat and barley, respectively. Crown Copyright (C) 2003 Published by Elsevier Ltd. All rights reserved. ER - TY - JFULL T1 - Beta2-adrenoceptor coupling to the Na/Ca exchanger depresses contraction in adult rat ventricular myocytes A1 - Sato, M A1 - Terracciano, CM A1 - Ranu, HK A1 - Harding, SE J1 - CIRCULATION Y1 - 2003/10/28/ VL - 108 SN - 0009-7322 SP - 179 EP - 180 ER - TY - JFULL T1 - Clinical recovery with successful LVAD explantation correlates with increased sarcoplasmic reticulum (SR) Ca content but not with regression of cellular hypertrophy A1 - Terracciano, CM A1 - Birks, E A1 - Hardy, J A1 - Harding, SE A1 - Tansley, P A1 - Barton, PJ A1 - Yacoub, MH J1 - CIRCULATION Y1 - 2003/10/28/ VL - 108 SN - 0009-7322 SP - 430 EP - 430 ER - TY - JFULL T1 - Chronic administration of clenbuterol affects ventricular myocyte size and calcium handling in rats A1 - Soppa, GK A1 - Malik, AH A1 - Terracciano, CM A1 - Yacoub, MH J1 - CIRCULATION Y1 - 2003/10/28/ VL - 108 SN - 0009-7322 SP - 331 EP - 331 ER - TY - JFULL T1 - N-acetyl aspartate estimation: a potential method for determining neuronal loss in the transmissible spongiform encephalopathies. A1 - Chung, YL A1 - Barr, J A1 - Bhakoo, K A1 - Williams, SC A1 - Bell, JD A1 - Fraser, JR J1 - Neuropathol Appl Neurobiol Y1 - 2003/10// VL - 29 SN - 0305-1846 SP - 445 EP - 450 N2 - Neurodegenerative pathology is typical of the transmissible spongiform encephalopathies (TSEs), and is thought to underlie clinical disease. Some morphometric studies have shown early focal neurone loss, but the full extent of TSE induced neuronal loss in the central nervous system is not known, and can only be accurately estimated using intensive morphometric techniques. We have used a murine scrapie model in which we determined the levels of N-acetyl aspartate (NAA), a putative neuronal marker, by both high-performance liquid chromatography and high resolution, proton magnetic resonance spectroscopy in samples taken sequentially from the hippocampus. This scrapie model develops severe neuronal loss in the hippocampus, and the NAA levels showed a significant positive correlation with our previous morphometric estimates of neurone number. NAA measurement may therefore provide a practical alternative to intensive morphometric techniques in the investigation of neurodegeneration in the TSEs. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14507336&query_hl=1 ER - TY - JFULL T1 - Chemokines acting via CXCR2 and CXCR4 control the release of neutrophils from the bone marrow and their return following senescence. A1 - Martin, C A1 - Burdon, PC A1 - Bridger, G A1 - Gutierrez-Ramos, JC A1 - Williams, TJ A1 - Rankin, SM J1 - Immunity Y1 - 2003/10// VL - 19 SN - 1074-7613 SP - 583 EP - 593 N2 - In this study we provide evidence that the SDF-1alpha/CXCR4 chemokine axis is involved in both the retention of neutrophils within the bone marrow and the homing of senescent neutrophils back to the bone marrow. We show that the functional responses of freshly isolated human and murine neutrophils to CXCR2 chemokines are significantly attenuated by SDF-1alpha, acting via CXCR4. As a consequence, the mobilization of neutrophils from the bone marrow in vivo by the CXCR2-chemokine, KC, was dramatically enhanced by blocking the effects of endogenous SDF-1alpha using a specific CXCR4 antagonist. As neutrophils age, they upregulate expression of CXCR4 and acquire the ability to migrate toward SDF-1alpha. We show here that these senescent CXCR4(high) neutrophils preferentially home to the bone marrow in vivo in a CXCR4-dependent manner, suggesting a previously undefined mechanism for the clearance of senescent neutrophils from the circulation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14563322&query_hl=1 ER - TY - JFULL T1 - Contractile effects of adenovirally-mediated increases in SERCA2a activity: a comparison between adult rat and rabbit ventricular myocytes. A1 - Chaudhri, B A1 - del Monte, F A1 - Hajjar, RJ A1 - Harding, SE J1 - Mol Cell Biochem Y1 - 2003/09// VL - 251 SN - 0300-8177 SP - 103 EP - 109 N2 - Adenoviral vectors have been successfully used to increase the activity of the sarcoplasmic reticulum Ca(2+)-ATPase in adult ventricular myocytes and to produce functional improvements in contractility in vivo and in vitro. While in vivo experiments are often performed in rat, in vitro manipulation of myocytes has been confined to rabbit and human cells. In the present study we make quantitative comparisons between cultured adult rat and rabbit myocytes in their responses to SERCA2a overexpression using adenoviral vectors. We also compare the strategy of SERCA2a overexpression with that of phospholamban down-regulation, using adenovirus carrying antisense message, as a means to increase SERCA2a activity and enhance contraction and relaxation. Adult myocytes were cultured for 48 h with either vector, and contraction assessed in 2 mM Ca2+, 37 degrees C, at a range of stimulation frequencies. Contraction amplitude was enhanced to a similar degree in either rat or rabbit myocytes at most stimulation frequencies, with SERCA2a overexpression and phospholamban down-regulation approximately equally effective. The maximum effect of either vector was less than that of beta-adrenoceptor agonists. Relaxation was accelerated in rabbit myocytes more strongly than in rat. Phospholamban antisense was slightly less effective than SERCA2a overexpression on relaxation times in rabbit. Increasing stimulation frequency also accelerated relaxation in rat myocytes: this effect was greater than, and additive with, that of SERCA2a overexpression. We conclude that, despite some species-dependent modification, the effects of increased SERCA2a activity are broadly similar in rat and rabbit. Both SERCA2a overexpression and phospholamban down-regulation are effective strategies, and neither appears to produce supraphysiological stimulatory effects on contraction or relaxation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14575311&query_hl=1 ER - TY - JFULL T1 - A novel approach for islet encapsulation with bio-membrane mimetic system A1 - Cui, W A1 - Barr, G A1 - Faucher, K A1 - Chaikof, E J1 - TRANSPLANTATION Y1 - 2003/08/27/ VL - 76 SN - 0041-1337 SP - S65 EP - S65 ER - TY - JFULL T1 - Nuclear receptors: a rendezvous for chromatin remodeling factors. A1 - Belandia, B A1 - Parker, MG J1 - Cell Y1 - 2003/08/08/ VL - 114 SN - 0092-8674 SP - 277 EP - 280 N2 - Nuclear receptors (NRs) are a large family of ligand-induced transcription factors that include the vitamin D receptor. The recent discovery of WINAC, a novel ATP-dependent chromatin remodeling complex, has shed new light on the molecular mechanisms by which the vitamin D receptor controls gene expression with unexpected clinical implications. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12914692&query_hl=1 ER - TY - JFULL T1 - Clinical heterogeneity in chronic myeloid leukaemia reflecting biological diversity in normal persons. A1 - Gordon, MY A1 - Marley, SB A1 - Apperley, JF A1 - Marin, D A1 - Kaeda, J A1 - Szydlo, R A1 - Goldman, JM J1 - Br J Haematol Y1 - 2003/08// VL - 122 SN - 0007-1048 SP - 424 EP - 429 N2 - The molecular basis of chronic myeloid leukaemia (CML) is well defined and highly consistent, yet prognosis varies considerably. This could reflect the biological diversity occurring in normal populations. We used a colony replating assay to measure the proliferative capacity of progenitor cells from 211 CML patients and 86 normal persons. Results were expressed as the frequency distributions of the proliferation index (PI) for individual cases. Normal PI values varied among individuals but were reproducible in individuals. The PIs for CML patients were moderately but significantly greater (P = 0.004) than normal values, consistent with increased progenitor cell proliferation in CML. Exposure of CML progenitor cells to the Abl-kinase inhibitor imatinib shifted their PI towards the normal range, implicating p210BCR-ABL. as a cause of the increased PI. The PIs of CML patients were higher than those of their human leucocyte antigen (HLA)-matched siblings PI (P = 0.003) and patient PI increased exponentially with sibling PI (r = 0.77; P = 0.001), but not with the PI values of HLA-matched unrelated individuals (P = 0.66). Finally, patients with high-risk prognostic scores (according to the Sokal or Hasford systems) had a significantly higher PI than those with low risk scores (P = 0.01 and 0.03 respectively). We conclude that heterogeneity in the CML patient population is analogous to the constitutional diversity in normal subjects. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12877669&query_hl=1 ER - TY - JFULL T1 - Chemokines in allergic airway disease. A1 - Lloyd, CM A1 - Rankin, SM J1 - Curr Opin Pharmacol Y1 - 2003/08// VL - 3 SN - 1471-4892 SP - 443 EP - 448 N2 - Expression of chemokine receptors on T helper 2 cells and eosinophils has been postulated to be the mechanism by which these cells are selectively recruited to the lung during allergic inflammatory reactions. Mouse models have provided evidence to show that blocking the ligands for these receptors is successful in abrogating the pathophysiological effects of allergen challenge. However, recent studies describing the effect of genetic deletions of these chemokine receptors have not confirmed the results obtained with ligand knockouts or neutralising antibodies. Coupled with the realisation that, because of a lack of species cross-reactivity, it is not possible to test small molecule antagonists against human receptors in the original in vivo animal models, the future of chemokine receptor therapeutics is in question. However, recent advances have been made regarding the therapeutic potential of blocking the chemokine receptors CCR3, CCR4 and CCR8 in allergic airway disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12901955&query_hl=1 ER - TY - JFULL T1 - Severe telomere shortening in patients with paroxysmal nocturnal hemoglobinuria affects both GPI- and GPI+ hematopoiesis. A1 - Karadimitris, A A1 - Araten, DJ A1 - Luzzatto, L A1 - Notaro, R J1 - Blood Y1 - 2003/07/15/ VL - 102 SN - 0006-4971 SP - 514 EP - 516 N2 - A most distinctive feature of paroxysmal nocturnal hemoglobinuria (PNH) is that in each patient glycosylphosphatidylinositol-negative (GPI-) and GPI+ hematopoietic stem cells (HSCs) coexist, and both contribute to hematopoiesis. Telomere size correlates inversely with the cell division history of HSCs. In 10 patients with hemolytic PNH the telomeres in sorted GPI- granulocytes were shorter than in sorted GPI+ granulocytes in 4 cases, comparable in 2 cases, and longer in the remaining 4 cases. Furthermore, the telomeres of both GPI- and GPI+ hematopoietic cells were markedly shortened compared with age-matched controls. The short telomeres in the GPI- cells probably reflect the large number of cell divisions required for the progeny of a single cell to contribute a large proportion of hematopoiesis. The short telomeres of the GPI+ cells indicate that the residual hematopoiesis contributed by these cells is not normal. This epigenetic change is an additional feature shared by PNH and aplastic anemia. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12623835&query_hl=1 ER - TY - JFULL T1 - Identification of RIP140 as a nuclear receptor cofactor with a role in female reproduction. A1 - Parker, M A1 - Leonardsson, G A1 - White, R A1 - Steel, J A1 - Milligan, S J1 - FEBS Lett Y1 - 2003/07/03/ VL - 546 SN - 0014-5793 SP - 149 EP - 153 N2 - Nuclear receptors function as ligand-dependent transcription factors by recruiting cofactors that remodel chromatin and recruit the transcription machinery. RIP140 (receptor interacting protein with a molecular weight of 140 kDa) is a widely expressed corepressor that has the potential to inhibit the transcriptional activity of most, if not all nuclear receptors. Mice devoid of RIP140 indicate that it plays a crucial role in female fertility and in adipose biology. It is essential in the ovary for ovulation, specifically oocyte release but not luteinisation of mature follicles. Our goal is to identify the key nuclear receptor(s) and their target genes responsible for ovulation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12829251&query_hl=1 ER - TY - JFULL T1 - Heritable gene silencing in lymphocytes delays chromatid resolution without affecting the timing of DNA replication. A1 - Azuara, V A1 - Brown, KE A1 - Williams, RR A1 - Webb, N A1 - Dillon, N A1 - Festenstein, R A1 - Buckle, V A1 - Merkenschlager, M A1 - Fisher, AG J1 - Nat Cell Biol Y1 - 2003/07// VL - 5 SN - 1465-7392 SP - 668 EP - 674 N2 - Temporal control of DNA replication has been implicated in epigenetic regulation of gene expression on the basis of observations that certain tissue-specific genes replicate earlier in expressing than non-expressing cells. Here, we show evidence that several leukocyte-specific genes replicate early in lymphocytes regardless of their transcription and also in fibroblasts, where these genes are never normally expressed. Instead, the heritable silencing of some genes (Rag-1, TdT, CD8alpha and lambda5) and their spatial recruitment to heterochromatin domains within the nucleus of lymphocytes resulted in a markedly delayed resolution of sister chromatids into doublet signals discernable by 3D fluorescence in situ hybridization (FISH). Integration of transgenes within heterochromatin (in cis) did, however, confer late replication and this was reversed after variegated transgene expression. These findings emphasise that chromosomal location is important for defining the replication timing of genes and show that retarded sister-chromatid resolution is a novel feature of inactive chromatin. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12833066&query_hl=1 ER - TY - JFULL T1 - Influence of culture and environmental conditions on the composition of exopolysaccharide produced by Agrobacterium radiobacter A1 - McKellar, RC A1 - van Geest, J A1 - Cui, W J1 - FOOD HYDROCOLLOID Y1 - 2003/07// VL - 17 SN - 0268-005X SP - 429 EP - 437 N2 - Exopolysaccharides (EPS) are produced by many genera of bacteria. These EPS have properties such as high viscosity, stabilizing and emusifying capabilites which make them attractive to the food industry. A strain of Agrobacterium radiobacter was isolated from lettuce which expressed a water-soluble acidic EPS, and a study of its properties was undertaken. EPS production was monitored using a dye binding assay with alcian blue, and concentration was expressed in xanthan equivalents (XE). Maximum production of EPS in mineral salts medium (MSM) occured after 3 days of growth at 28 degreesC. Both the lettuce isolate (FRP 695) and A. radiobacter ATTCC 6466 (designated FRP 718) produced EPS composed mainly of glucose and galactose. When grown with yeast extract (YE), however, both strains produced EPS containing mannose. The mannose content of the EPS was proportional to YE concentration up to 0.35%, but the yield of EPS was not significantly (P > 0.05) influenced by YE. In seed medium containing both YE and peptone, neither the inoculum level (0.25-5.0%) nor pH (5.5-8.0) nor sucrose concentration (0.25-10%) had any significant (P > 0.05) effect on the EPS yield. The mannose content was influenced by decreasing sucrose in seed medium, with a maximum of 78% and a minimum of 28% at 0.25 and 7.5% sucrose, respectively. Over all experiment conditions (n = 50), mannose content only exceeded 25% (% dry weight) in seed medium; mannose content was less than or equal to23% when the basal medium was MSM. The results suggest that useful changes in composition of EPS can be made through manipulation of the growth environment. Crown Copyright (C) 2003 Published by Elsevier Science Ltd. All rights reserved. ER - TY - JFULL T1 - Changes in sarcolemmal Ca entry and sarcoplasmic reticulum Ca content in ventricular myocytes from patients with end-stage heart failure following myocardial recovery after combined pharmacological and ventricular assist device therapy. A1 - Terracciano, CM A1 - Harding, SE A1 - Adamson, D A1 - Koban, M A1 - Tansley, P A1 - Birks, EJ A1 - Barton, PJ A1 - Yacoub, MH J1 - Eur Heart J Y1 - 2003/07// VL - 24 SN - 0195-668X SP - 1329 EP - 1339 N2 - AIMS: Support with left ventricular assist devices (LVAD) improves cardiac performance in patients with end-stage heart failure. In some cases this strategy, combined with pharmacological treatment, has led to a clinical improvement which remained after LVAD explant. This study defines changes in Ca handling at the cellular level in failing left ventricular tissue taken at LVAD implant (LVAD core) and LVAD removal (post-LVAD). METHODS AND RESULTS: We studied cell size and Ca regulation in enzymatically dissociated cardiac myocytes. We used confocal microscopy and electrophysiological techniques to investigate the SR Ca content and major Ca movements across the sarcolemma during the action potential. We firstly recorded a significant reduction in cell capacitance and cell volume consistent with regression of cellular hypertrophy in post-LVAD myocytes compared with LVAD core myocytes. Ca entry via sarcolemmal Ca channels during the action potential using action potential voltage-clamping was significantly increased in post-LVAD myocytes compared with LVAD cores myocytes. Finally, SR Ca content (assessed by integrating the caffeine-induced Na/Ca exchanger transient inward current) in post-LVAD myocytes was also significantly increased compared with LVAD cores myocytes. CONCLUSIONS: These results show that in myocytes from patients after LVAD support there is more Ca entry to trigger Ca release and more SR Ca content, leading to improved contractile function. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12871690&query_hl=1 ER - TY - JFULL T1 - Phosphodiesterase type 5 as a target for the treatment of hypoxia-induced pulmonary hypertension. A1 - Sebkhi, A A1 - Strange, JW A1 - Phillips, SC A1 - Wharton, J A1 - Wilkins, MR J1 - Circulation Y1 - 2003/07/01/ VL - 107 SN - 1524-4539 SP - 3230 EP - 3235 N2 - BACKGROUND: Phosphodiesterase type 5 (PDE5) is a novel therapeutic target for the treatment of pulmonary hypertension. This study examined the distribution of PDE5 in normal and hypoxic lung and the effect of chronic PDE5 inhibition with sildenafil, initiated before and during exposure to hypoxia, on pulmonary artery pressure (PAP) and structure. METHODS AND RESULTS: Sprague-Dawley rats were exposed to hypoxia (10% O2) for up to 42 days. PAP, measured continuously by telemetry, increased gradually by 20 to 40 mm Hg, reaching a plateau between 10 and 14 days, and declined to normal levels on return to normoxia. PDE5 immunoreactivity was localized to smooth muscle cells in the medial layer of pulmonary arteries and veins in the normal lung and in distal muscularized arteries (<25 microm diameter) after hypoxia-induced pulmonary hypertension. Sildenafil (25 or 75 mg x kg(-1) x d(-1)) given before hypoxia produced marked dose-dependent inhibition in the rise of PAP (60% to 90% reduction; P<0.0001) and vascular muscularization (28.4+/-5.0% reduction; P<0.001). When begun after 14 days of hypoxia, sildenafil significantly reduced PAP (30% reduction; P<0.0001) and partially reversed pulmonary artery muscularization (39.9+/-4.9% reduction; P<0.001). CONCLUSIONS: PDE5 is found throughout the muscularized pulmonary vascular tree, including in newly muscularized distal pulmonary arteries exposed to hypoxia. PDE5 inhibition attenuates the rise in PAP and vascular remodeling when given before chronic exposure to hypoxia and when administered as a treatment during ongoing hypoxia-induced pulmonary hypertension. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12796132&query_hl=1 ER - TY - JFULL T1 - Telomerase-immortalized sheep fibroblasts can be reprogrammed by nuclear transfer to undergo early development. A1 - Cui, W A1 - Wylie, D A1 - Aslam, S A1 - Dinnyes, A A1 - King, T A1 - Wilmut, I A1 - Clark, AJ J1 - Biol Reprod Y1 - 2003/07// VL - 69 SN - 0006-3363 SP - 15 EP - 21 N2 - Telomere shortening and lack of telomerase activity have been implicated in cellular senescence in human fibroblasts. Expression of the human telomerase catalytic reverse transcriptase subunit (hTERT) in these cells reconstitutes telomerase activity and immortalizes the cells without tumor transformation. In this report, we show that sheep fibroblasts are similar to human cells. They do not have detectable telomerase activity and undergo only a finite numbers of cell divisions before replicative senescence. Telomere lengths in sheep fibroblasts are similar to those reported for human cells and shorten at a rate of 50-200 base pairs (bp) each cell division. Expression of the human telomerase catalytic subunit restored the telomerase activity in the sheep cells and extended their proliferative life span. None of the telomerase positive sheep fibroblasts exhibited a transformed phenotype after 200 days of continuous culture, and the higher hTERT expressing cells maintained their telomere lengths and normal cell characteristics for more than 500 days in culture. In cloning experiments using one of these cell lines as a nuclear donor, the reconstructed karyoplasts were reprogrammed and developed to the blastocyst stage at a similar frequency to that observed with the parental, telomerase negative cell line. After embryo transfer the blastocysts exhibited a relatively high frequency of implantation, early fetal development, and organogenesis. No fetuses survived beyond 40 days of development, however, showing that although these cells could be substantially reprogrammed, they were not fully competent for nuclear transfer. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12606403&query_hl=1 ER - TY - JFULL T1 - Differential regulation of human eosinophil IL-3, IL-5, and GM-CSF receptor alpha-chain expression by cytokines: IL-3, IL-5, and GM-CSF down-regulate IL-5 receptor alpha expression with loss of IL-5 responsiveness, but up-regulate IL-3 receptor alpha expression. A1 - Gregory, B A1 - Kirchem, A A1 - Phipps, S A1 - Gevaert, P A1 - Pridgeon, C A1 - Rankin, SM A1 - Robinson, DS J1 - J Immunol Y1 - 2003/06/01/ VL - 170 SN - 0022-1767 SP - 5359 EP - 5366 N2 - Our recent data suggested that tissue eosinophils may be relatively insensitive to anti-IL-5 treatment. We examined cross-regulation and functional consequences of modulation of eosinophil cytokine receptor expression by IL-3, IL-5 GM-CSF, and eotaxin. Incubation of eosinophils with IL-3, IL-5, or GM-CSF led to reduced expression of IL-5R alpha, which was sustained for up to 5 days. Eosinophils incubated with IL-5 or IL-3 showed diminished respiratory burst and mitogen-activated protein kinase kinase phosphorylation in response to further IL-5 stimulation. In contrast to these findings, eosinophil expression of IL-3R alpha was increased by IL-3, IL-5, and GM-CSF, whereas GM-CSF receptor alpha was down-regulated by GM-CSF, but was not affected by IL-3 or IL-5. CCR3 expression was down-regulated by IL-3 and was transiently reduced by IL-5 and GM-CSF, but rapidly returned toward baseline. Eotaxin had no effect on receptor expression for IL-3, IL-5, or GM-CSF. Up-regulation of IL-3R alpha by cytokines was prevented by a phosphoinositol 3-kinase inhibitor, whereas this and other signaling inhibitors had no effect on IL-5R alpha down-regulation. These data suggest dynamic and differential regulation of eosinophil receptors for IL-3, IL-5, and GM-CSF by the cytokine ligands. Since these cytokines are thought to be involved in eosinophil development and mobilization from the bone marrow and are present at sites of allergic inflammation, tissue eosinophils may have reduced IL-5R expression and responsiveness, and this may explain the disappointing effect of anti-IL-5 therapy in reducing airway eosinophilia in asthma. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12759409&query_hl=1 ER - TY - JFULL T1 - Scaffolds and biomaterials for tissue engineering: a review of clinical applications. A1 - Vats, A A1 - Tolley, NS A1 - Polak, JM A1 - Gough, JE J1 - Clin Otolaryngol Allied Sci Y1 - 2003/06// VL - 28 SN - 0307-7772 SP - 165 EP - 172 N2 - Tissue engineering is a multidisciplinary area of research aimed at regeneration of tissues and restoration of organ function. This is achieved through implantation of cells/tissues grown outside the body or by stimulating cells to grow into an implanted matrix. In this short review, we discuss the use of biomaterials, in the form of scaffolds, for tissue engineering and review clinical applications to otorhinolaryngology-head and neck surgery. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12755749&query_hl=1 ER - TY - JFULL T1 - Proliferative lifespan is conserved after nuclear transfer. A1 - Clark, AJ A1 - Ferrier, P A1 - Aslam, S A1 - Burl, S A1 - Denning, C A1 - Wylie, D A1 - Ross, A A1 - de Sousa, P A1 - Wilmut, I A1 - Cui, W J1 - Nat Cell Biol Y1 - 2003/06// VL - 5 SN - 1465-7392 SP - 535 EP - 538 N2 - Cultured primary cells exhibit a finite proliferative lifespan, termed the Hayflick limit. Cloning by nuclear transfer can reverse this cellular ageing process and can be accomplished with cultured cells nearing senescence. Here we describe nuclear transfer experiments in which donor cell lines at different ages and with different proliferative capacities were used to clone foetuses and animals from which new primary cell lines were generated. The rederived lines had the same proliferative capacity and rate of telomere shortening as the donor cell lines, suggesting that these are innate, genetically determined, properties that are conserved by nuclear transfer. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12738962&query_hl=1 ER - TY - JFULL T1 - Gene expression: Oestrogen receptor hijacked. A1 - Brosens, JJ A1 - Parker, MG J1 - Nature Y1 - 2003/05/29/ VL - 423 SN - 0028-0836 SP - 487 EP - 488 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12774104&query_hl=1 ER - TY - JFULL T1 - Generation of CD1 tetramers as a tool to monitor glycolipid-specific T cells. A1 - Gadola, SD A1 - Karadimitris, A A1 - Zaccai, NR A1 - Salio, M A1 - Dulphy, N A1 - Shepherd, D A1 - Jones, EY A1 - Cerundolo, V J1 - Philos Trans R Soc Lond B Biol Sci Y1 - 2003/05/29/ VL - 358 SN - 0962-8436 SP - 875 EP - 877 N2 - CD1 molecules are beta(2)m-associated HLA class-I-like glycoproteins which have the unique ability to present glycolipid and phospholipid antigens to specific T lymphocytes. To study the biology of CD1 and its role in human disease we developed novel techniques for generation of recombinant CD1/lipid complexes by in vitro refolding. Fluorescent tetrameric complexes made from soluble recombinant CD1d/alpha-galactosylceramide complexes allowed highly sensitive and specific ex vivo and in vitro detection and functional characterization of novel human T-lymphocyte populations. Furthermore, protein crystals were obtained from soluble recombinant CD1b/beta(2)m-proteins loaded either with phosphatidylinositol or ganglioside GM2, which led to the first atomic structure determination of a CD1/lipid complex. The analysis of these crystal structures clarified how CD1b molecules can bind lipid ligands of different size, and revealed a broader spectrum of potential CD1b ligands than previously predicted. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12803920&query_hl=1 ER - TY - JFULL T1 - Prospective evaluation of iNOS and oxidant stress in the first three months after cardiac transplantation; Relationship to acute rejection and myocardial function A1 - Birks, EJ A1 - Yacoub, MH A1 - Bishop, AE A1 - Burke, M A1 - Khaghani, A A1 - Polak, JM A1 - Banner, NR J1 - HEART Y1 - 2003/05// VL - 89 SN - 1355-6037 SP - A24 EP - A24 ER - TY - JFULL T1 - Progenitor cells divide symmetrically to generate new colony-forming cells and clonal heterogeneity. A1 - Marley, SB A1 - Lewis, JL A1 - Gordon, MY J1 - Br J Haematol Y1 - 2003/05// VL - 121 SN - 0007-1048 SP - 643 EP - 648 N2 - Self-renewal is the most fundamental property of haemopoietic stem and progenitor cells. However, because of the need to produce differentiated cells, not all cell divisions involve self-renewal. We have used a colony replating assay to follow the fates of individual haemopoietic progenitor cell clones. For this, human myeloid colony-forming cells (CFCs) were cultured by standard methodology. Onset of proliferation and growth rates were established by a video recording method. Individual colonies were replated several times to document the rate of clonal extinction, and the numbers of secondary, tertiary and quaternary CFCs. The clonogenic population exhibited similar kinetics in terms of onset of proliferation and growth rate. Clonal extinction was progressive so that only 30 +/- 7% (mean +/- standard error of the mean; n = 4) of the original primary colonies formed quaternary colonies after the third replating step. However, individual primary CFCs that produced colonies throughout the experiment generated, on average, 40 +/- 8 secondary and tertiary CFCs overall. The values obtained in standard culture conditions were modified when granulocyte colony-stimulating factor (G-CSF) or G-CSF plus interleukin 3 were used to stimulate colony growth, showing that the kinetics of colony formation respond to extrinsic regulation. Examination of the replating potential of individual secondary colonies in the clones demonstrated that they generated different numbers of tertiary colonies. The data best fit a stochastic model of haemopoietic cell development where event probabilities can be modified by extracellular factors. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12752107&query_hl=1 ER - TY - JFULL T1 - Mafs, Pax6 and Prox1 cooperate to regulate bB1 crystallin gene expression in developing lens A1 - Cui, W A1 - Tomarev, SI A1 - Chepelinsky, AB A1 - Duncan, MK J1 - INVEST OPHTH VIS SCI Y1 - 2003/05// VL - 44 SN - 0146-0404 SP - U290 EP - U290 ER - TY - JFULL T1 - Changes in gene expression in response to mechanical strain in human scleral fibroblast A1 - Cui, W A1 - Bryant, M A1 - McDonnell, PJ J1 - INVEST OPHTH VIS SCI Y1 - 2003/05// VL - 44 SN - 0146-0404 SP - U55 EP - U55 ER - TY - JFULL T1 - Adenovirally-overexpressed beta 1- and beta 2-adrenoceptors enhances the contractile response to CGP 12177A in adult rat cardiomyocytes A1 - Lewis, CJ A1 - Koch, WJ A1 - Brown, MJ A1 - Harding, SE J1 - BRIT J PHARMACOL Y1 - 2003/04// VL - 138 SN - 0007-1188 SP - U9 EP - U9 ER - TY - JFULL T1 - Analysis of bile in patients with and without pancreaticobiliary malignancy by in vitro 31-phosphorus NMR spectroscopy A1 - Khan, SA A1 - Cox, IJ A1 - Bansi, D A1 - Thillainayagam, A A1 - Thomas, HC A1 - Taylor-Robinson, SD J1 - GUT Y1 - 2003/04// VL - 52 SN - 0017-5749 SP - A92 EP - A92 ER - TY - JFULL T1 - Birth medical risk status, infant mortality and preschool morbidity A1 - Roth, J A1 - Cui, W A1 - Wu, S A1 - Ariet, M A1 - Ma, CX A1 - Resnick, MB A1 - Morse, SB J1 - PEDIATR RES Y1 - 2003/04// VL - 53 SN - 0031-3998 SP - 458A EP - 458A ER - TY - JFULL T1 - Novel p53 mutations but lack of a mutational fingerprint in human intrahepatic cholangiocarcinoma A1 - Khan, SA A1 - Taylor-Robinson, SD A1 - Carmichael, PL A1 - Habib, N A1 - Lemoine, N A1 - Thomas, HC J1 - GUT Y1 - 2003/04// VL - 52 SN - 0017-5749 SP - A32 EP - A32 ER - TY - JFULL T1 - DNA adducts, detected by 32P postlabelling, in human cholangiocarcinoma. A1 - Khan, SA A1 - Carmichael, PL A1 - Taylor-Robinson, SD A1 - Habib, N A1 - Thomas, HC J1 - Gut Y1 - 2003/04// VL - 52 SN - 0017-5749 SP - 586 EP - 591 N2 - BACKGROUND: Reported mortality from intrahepatic cholangiocarcinoma (CCa) has risen steeply in the UK and other industrialised countries over the past 30 years, the cause of which has not been explained. DNA adduct formation is promutagenic and demonstrates exposure to a DNA damaging agent. It is a key step in chemically induced carcinogenesis. We hypothesise that the increase in CCa mortality is caused by a rise in a genotoxic environmental agent(s), causing cholangiocyte DNA damage. AIMS: To investigate and compare tumour and tumour adjacent CCa tissue, and non-cancer control bile duct tissue, for DNA adducts as a biomarker of genotoxin exposure. METHODS: DNA from 32 intrahepatic CCa patients (and in 28 cases DNA from adjacent non-tumour tissue) and from biliary ducts of seven non-cancer patients were investigated for the presence of DNA adducts using the nuclease P1 method of (32)P postlabelling. DNA adduct levels (number of adducts/10(8) nucleotides) were quantified. RESULTS: There was no significant difference in relative adduct labellings (RALs) between tumour adjacent DNA (median 8.6, range 1.2-51.6) and CCa DNA (7.2, 1.8-48.4). However, RALs were significantly higher in DNA from cancer patients (tumour adjacent and CCa DNA) compared with non-cancer patient DNA (2.9, 0.6-11.5; p=0.032, two tailed Mann-Whitney U test). Different adduct patterns were also seen in CCa compared with non-cancer patients. CONCLUSION: Quantitative and qualitative differences in adducts between cancer and non-cancer patients support the hypothesis that genotoxins may play a role in the development of intrahepatic CCa. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12631674&query_hl=1 ER - TY - JFULL T1 - Targeting urothelium: ex vivo assay standardization and selection of internalizing ligands A1 - Ardelt, PU A1 - Wood, CG A1 - Chen, L A1 - Mintz, PJ A1 - Moya, C A1 - Arap, MA A1 - Wright, KC A1 - Pasqualini, R A1 - Arap, W J1 - J Urol. Y1 - 2003/04// IS - 4 VL - 169 SN - 0022-5347 SP - 1535 EP - 1540 ER - TY - JFULL T1 - Cardiostimulant and cardiodepressant effects through overexpressed human beta(2)-adrenoceptors in murine heart: regional differences and functional role of beta(1)-adrenoceptors A1 - Heubach, JF A1 - Blaschke, M A1 - Harding, SE A1 - Ravens, U A1 - Kaumann, AJ J1 - N-S ARCH PHARMACOL Y1 - 2003/04// VL - 367 SN - 0028-1298 SP - 380 EP - 390 N2 - (-)-Isoprenaline enhances cardiac contractility through beta-adrenoceptors. However, in cardiac tissue from transgenic mice with a 200-400-fold cardiac overexpression of the human beta(2)-adrenoceptor (TG4) we observed a pronounced cardiodepression at high (-)-isoprenaline concentrations. Here, we investigated the functional role of the coexisting beta(1), beta(2), and beta(3)-adrenoceptor subtypes in several regions of the TG4 heart, and in particular their contribution to the negative inotropic effect. In paced TG4 left atria, (-)-isoprenaline produced bell-shaped concentration-effect curves increasing (-logEC(50)M=9.0) and decreasing (-logIC(50)M=6.4) contractile force. These effects were unaffected by the beta(1)-selective CGP 20712A (300 nM). The beta(2)-selective inverse agonist ICI 118,551 (30-1.000 nM) antagonised in surmountable manner both the positive and negative inotropic effects of (-)-isoprenaline with similar concentration-dependence, consistent with an exclusive mediation through beta(2)-adrenoceptors. The beta(3)-adrenoceptor-selective agonist BRL37344 (1 nM-10 muM) failed to produce significant inotropic effects in TG4 left atria. Subsequently, we measured left atrial action potentials accompanying the inotropic changes induced by (-)-isoprenaline. Action potentials tended to have shorter duration in left atria from TG4 mice than from non-transgenic littermate mice, However, (-)-isoprenaline prolonged the duration of 30% repolarisation in atria from non-transgenic littermate but not from TG4 mice, while 90% repolarisation was abbreviated in both groups of atria. Negative inotropic effects of (-)-isoprenaline were also observed in right ventricular preparations. Pertussis toxin-treatment of the mice abolished the negative inotropic effects in left atria and reduced cardiodepression in right ventricle, indicating an involvement of beta(2)-adrenoceptor coupling to PTX-sensitive G-proteins. In additional experiments, designed to study the native murine beta(1)-adrenoceptor function, we used the physiological beta(1)-adrenoceptor agonist (-)-noradrenaline. In the presence of 600 nM ICI 118,551 we failed to find a functional role of the beta(1)-adrenoceptors in left atria, and detected only a marginal contribution to the positive chronotropic effect in right atria. We also investigated the effects of the non-conventional partial agonist (-)-CGP 12177 (0.2 nM-6 muM), which in wild-type mice causes tachycardia through beta(1)-adrenoceptors. In TG4 right atria, however, (-)-CGP 12177-evoked tachycardia was resistant to blockade by CGP 20712A but antagonised by ICI 118,551, consistent with mediation, through human beta(2)-adrenoceptors.The results from TG4 mice suggest that the positive and negative inotropic effects of (-)-isoprenaline are mediated through human overexpressed beta(2)-adrenoceptors coupled to G(s) protein and G(i) protein, respectively. Them (-)-isoprenaline-evoked shortening of the atrial action potential combined with reduced responses of L-type Ca2+ current may contribute to the negative inotropic effects. The function of marine cardiac beta(1)-adrenoceptors is suppressed by overexpressed human beta(2)-adrenoceptors. ER - TY - JFULL T1 - The optimal slice thickness for left ventricular volume calculation A1 - Cui, W A1 - Kondo, T A1 - Sato, T A1 - Anno, H A1 - Yoshihiro, I A1 - Sarai, M A1 - Shinozaki, H A1 - Kakizawa, S A1 - Sugiura, K A1 - Oshima, K A1 - Katada, K A1 - Hishida, H J1 - J AM COLL CARDIOL Y1 - 2003/03/19/ VL - 41 SN - 0735-1097 SP - 422A EP - 422A ER - TY - JFULL T1 - Chronic myeloid leukemia in chronic phase responding to imatinib: the occurrence of additional cytogenetic abnormalities predicts disease progression. A1 - Marktel, S A1 - Marin, D A1 - Foot, N A1 - Szydlo, R A1 - Bua, M A1 - Karadimitris, A A1 - De Melo, VA A1 - Kotzampaltiris, P A1 - Dazzi, F A1 - Rahemtulla, A A1 - Olavarria, E A1 - Apperley, JF A1 - Goldman, JM J1 - Haematologica Y1 - 2003/03// VL - 88 SN - 0390-6078 SP - 260 EP - 267 N2 - BACKGROUND AND OBJECTIVES: The acquisition of additional cytogenetic changes (clonal evolution, CE) during treatment of chronic myeloid leukemia (CML) with imatinib mesylate is currently regarded as an index of increasing resistance to imatinib. Therefore, to investigate whether CE as an isolated event increases the risk of disease progression during imatinib treatment, we compared the outcome of patients with CML in chronic phase (CML-CP) who developed CE whilst in complete hematologic remission with the outcome of comparable patients in complete hematologic remission who showed no evidence of CE. DESIGN AND METHODS: We serially studied cytogenetic findings in 102 patients receiving the Abl-tyrosine kinase inhibitor, imatinib mesylate, as sole agent to treat CML-CP who had no evidence of CE before initiation of imatinib treatment. RESULTS: CE was identified during treatment with imatinib in 15 patients, 10 of whom were in complete hematologic remission. In most cases these changes occurred exclusively in the Ph+ population but in three patients additional changes occurred in a co-existing Ph-negative population. Patients with de novo CE in the absence of any other sign of disease progression had a significantly higher incidence of progression by 18 months than did non-CE patients (progression-free survival 34.3% (CI 10.5-69.8%) vs. 94.1% (CI 80.6-98.4%), p<0.0001). INTERPRETATION AND CONCLUSIONS: Based on this relatively small series of patients, we conclude that acquisition of clonal evolution increases the risk of subsequent disease progression also in CML patients in complete hematologic remission on imatinib. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12651263&query_hl=1 ER - TY - JFULL T1 - Interstitial vascularity in fibrosing alveolitis. A1 - Renzoni, EA A1 - Walsh, DA A1 - Salmon, M A1 - Wells, AU A1 - Sestini, P A1 - Nicholson, AG A1 - Veeraraghavan, S A1 - Bishop, AE A1 - Romanska, HM A1 - Pantelidis, P A1 - Black, CM A1 - Du Bois, RM J1 - Am J Respir Crit Care Med Y1 - 2003/02/01/ VL - 167 SN - 1073-449X SP - 438 EP - 443 N2 - The aim of this study was to evaluate interstitial vascularity in cryptogenic fibrosing alveolitis (CFA) and in fibrosing alveolitis associated with systemic sclerosis (FASSc). Open lung biopsies from eight patients with CFA, nine patients with FASSc, and normal lung from 12 patients undergoing surgery for lung cancer were studied. Markers for endothelial cells (CD34) and cell proliferation (proliferating cell nuclear antigen) were localized by sequential immunohistochemistry and quantified using computer-assisted analysis. Vascular distribution was evaluated at increasing distances (up to 160 microm) from the airspaces. Vessel density was markedly reduced in both FASSc (3.9%) and in CFA (4.5%) compared with control samples (20.4%, p < 0.0001). The percentage of tissue occupied by vessels decreased with increasing distance from alveoli in control samples but not in CFA or FASSc samples. Endothelial cell proliferation indices were increased in FASSc but not in CFA, compared with control samples (p = 0.006). In conclusion, there is net vascular ablation and redistribution of blood vessels in areas of interstitial thickening in both CFA and FASSc, which may contribute to gas exchange impairment. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12406847&query_hl=1 ER - TY - JFULL T1 - In vivo monitoring of rat brain metabolites during vigabatrin treatment using localized 2D-COSY. A1 - Welch, JW A1 - Bhakoo, K A1 - Dixon, RM A1 - Styles, P A1 - Sibson, NR A1 - Blamire, AM J1 - NMR Biomed Y1 - 2003/02// VL - 16 SN - 0952-3480 SP - 47 EP - 54 N2 - A two-dimensional COSY-based localization sequence was designed to allow the in vivo monitoring of proton metabolites in rat brain [particularly gamma-aminobutyric acid (GABA), glutamine, taurine and myo-inositol]. The sequence incorporated OSIRIS signal localization, B1-insensitive water suppression and phase-sensitive COSY acquisition. The method was used to study the effects of the GABA-transaminase inhibitor vigabatrin on rat brain metabolite concentrations. Wistar rats were treated daily for 3 days with an oral dose of vigabatrin (200 mg/kg, n = 4). Localized COSY spectra were obtained during a 120 min acquisition from a 270 microl central brain voxel and compared with nine untreated control animals. Significant elevations were observed in GABA (267% of control, p < 0.005, Mann-Witney test), glutamine (130% of control, p < 0.005) and taurine (113% of control, p < 0.05). Changes in GABA and taurine were consistent with previous data on the action of Vigabatrin, and support a previously hypothesized link between these compounds. The increase in glutamine was more surprising and may reflect the balance between the level and/or site of GABA-transaminase inhibition and downregulation of GABA synthesis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12577297&query_hl=1 ER - TY - JFULL T1 - Pulmonary arterial hypertension and the vasoconstrictive factor: is there still a role for vasodilator testing? A1 - Gibbs, JS A1 - Wharton, J A1 - Wilkins, MR J1 - Eur Heart J Y1 - 2003/02// VL - 24 SN - 0195-668X SP - 297 EP - 298 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12581676&query_hl=1 ER - TY - JFULL T1 - Adenovirally-overexpressed beta(1)ARs activate contraction by a non-cyclic AMP-dependent pathway in adult rat myocytes A1 - Gong, HB A1 - Lewis, CJ A1 - Terracciano, CMN A1 - Koch, WJ A1 - O'Gara, P A1 - Brown, MJ A1 - Harding, SE J1 - BIOPHYS J Y1 - 2003/02// VL - 84 SN - 0006-3495 SP - 397A EP - 397A ER - TY - JFULL T1 - Frequency and phenotype of circulating Valpha24/Vbeta11 double-positive natural killer T cells during hepatitis C virus infection. A1 - Lucas, M A1 - Gadola, S A1 - Meier, U A1 - Young, NT A1 - Harcourt, G A1 - Karadimitris, A A1 - Coumi, N A1 - Brown, D A1 - Dusheiko, G A1 - Cerundolo, V A1 - Klenerman, P J1 - J Virol Y1 - 2003/02// VL - 77 SN - 0022-538X SP - 2251 EP - 2257 N2 - Natural killer T (NKT) cells are thought to be involved in innate responses against infection. We investigated one specific type of NKT cell, Valpha24/Vbeta11 double positive, in hepatitis C virus (HCV) infection. Lower frequencies of this population were detected in the blood of HCV PCR-positive patients than in controls. Unlike Valpha24/Vbeta11 NKT cells found in blood, those in the liver appeared to be recently activated. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12525661&query_hl=1 ER - TY - JFULL T1 - A tumour that secretes glucagon-like peptide-1 and somatostatin in a patient with reactive hypoglycaemia and diabetes. A1 - Todd, JF A1 - Stanley, SA A1 - Roufosse, CA A1 - Bishop, AE A1 - Khoo, B A1 - Bloom, SR A1 - Meeran, K J1 - Lancet Y1 - 2003/01/18/ VL - 361 SN - 0140-6736 SP - 228 EP - 230 N2 - Glucagon-like peptide 1 (GLP-1), an insulinotropic hormone normally synthesised in the intestinal mucosa and released in response to a meal, is essential for normal glucose homoeostasis. There is much interest in the use of GLP-1 to treat diabetes, since the risk of hypoglycaemia is thought to be low. We report an instance of a 45-year-old woman with a GLP-1 and somatostatin secreting neuroendocrine tumour who presented with reactive hypoglycaemia and hyperglycaemia, but who was subsequently cured by surgery. This case, of a neuroendocrine tumour secreting GLP-1 and causing reactive hypoglycaemia, indicates a potential adverse effect of GLP-1 therapy for diabetes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12547550&query_hl=1 ER - TY - JFULL T1 - Unfriendly chemicals in pancreatic cancer - Invited commentary A1 - Khan, SA A1 - Beck, A A1 - Carmichael, PL A1 - Taylor-Robinson, SD J1 - PANCREATOLOGY Y1 - 2003/// VL - 3 SN - 1424-3903 SP - 7 EP - 8 ER - TY - JFULL T1 - Spectroscopic study of human lung epithelial cells (A549) in culture: living cells versus dead cells. A1 - Notingher, I A1 - Verrier, S A1 - Haque, S A1 - Polak, JM A1 - Hench, LL J1 - Biopolymers Y1 - 2003/// VL - 72 SN - 0006-3525 SP - 230 EP - 240 N2 - The noninvasive analysis of living cells grown on 3-dimensional scaffold materials is a key point in tissue engineering. In this work we show the capability of Raman spectroscopy for use as a noninvasive method to distinguish cells at different stages of the cell cycle and living cells from dead cells. The spectral differences between cells in different stages of the cell cycle are characterized mainly by variations in DNA vibrations at 782, 788, and 1095 cm(-1). The Raman spectrum of dead human lung derived (A549 line) cells indicates the breakdown of both phosphodiester bonds and DNA bases. The most sensitive peak for identifying dead cells is the 788 cm(-1) peak corresponding to DNA Obond;Pbond;O backbone stretching. The magnitude of this peak is reduced by 80% in the spectrum of dead cells. Changes in protein peaks suggest significant conformational changes; for example, the magnitude of the 1231 cm(-1) peak assigned to random coils is reduced by 63% for dead cells. The sharp peak of phenylalanine at 1005 cm(-1) drops to half, indicating a decrease of stable proteins associated with cell death. The differences in the 1190-1385 cm(-1) spectral region also suggest a decrease in the amount of nucleic acids and proteins. Using curve fitting, we quantify these spectral differences that can be used as markers of cell death. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12833477&query_hl=1 ER - TY - JFULL T1 - Applicationi of FTIR and Raman spectroscopy to characterisation of bioactive materials and living cells A1 - Notingher I A1 - Jones JR A1 - Verrier S A1 - Bisson I A1 - Embanga PP A1 - Edwards P A1 - Polak JM A1 - Hench LL J1 - Spectroscopy: An International Journal Y1 - 2003/// VL - 17 SP - 275 EP - 278 ER - TY - JFULL T1 - Osteoclast formation from circulating precursors in osteoporosis. A1 - Jevon, M A1 - Hirayama, T A1 - Brown, MA A1 - Wass, JA A1 - Sabokbar, A A1 - Ostelere, S A1 - Athenasou, NA J1 - Scand J Rheumatol Y1 - 2003/// VL - 32 SN - 0300-9742 SP - 95 EP - 100 N2 - OBJECTIVE: An imbalance between bone formation and bone resorption is thought to underlie the pathogenesis of reduced bone mass in osteoporosis. Bone resorption is carried out by osteoclasts. which are formed from marrow-derived cells that circulate in the monocyte fraction. The aim of this study was to determine the role of osteoclast formation in the pathogenesis of bone loss in osteoporosis. METHODS: The proportion of circulating osteoclast precursors and their relative sensitivity to the osteoclastogenic effects of M-CSF. 1,25(OH)2D3 and RANKL were assessed in primary osteoporosis patients and normal controls. RESULTS: Although there was no difference in the number of circulating osteoclast precursors in osteoporosis patients and normal controls. osteoclasts formed from osteoporosis patients exhibited substantially increased resorptive activity relative to normal controls. Although no increased sensitivity to the osteoclastogenic effects of 1,25(OH)D3 or M-CSF was noted, increased bone resorption was found in osteoporosis peripheral blood mononuclear cell (PBMC) cultures to which these factors were added. CONCLUSION: Our findings suggest that osteoclast functional activity rather than formation is increased in primary involutional osteoporosis and that dexamethasone acts to increase osteoclast formation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12737328&query_hl=1 ER - TY - JFULL T1 - Brain damage results in down-regulation of N-acetylaspartate as a neuronal osmolyte. A1 - Baslow, MH A1 - Suckow, RF A1 - Gaynor, K A1 - Bhakoo, KK A1 - Marks, N A1 - Saito, M A1 - Saito, M A1 - Duff, K A1 - Matsuoka, Y A1 - Berg, MJ J1 - Neuromolecular Med Y1 - 2003/// VL - 3 SN - 1535-1084 SP - 95 EP - 104 N2 - N-acetyl-L-aspartate (NAA) is present in the vertebrate brain, where its concentration is one of the highest of all free amino acids. Although NAA is synthesized and stored primarily in neurons, it is not hydrolyzed in these cells. However, after its regulated release into extracellular fluid, neuronal NAA is hydrolyzed by amidohydrolase II that is present in oligodendrocytes. About 30% of neurons do not contain appreciable amounts of NAA, but its prominence in 1H nuclear magnetic resonance spectroscopic (MRS) studies has led to its wide use as a neuronal marker in diagnostic human medicine as both an indicator of brain pathology, and of disease progression in a variety of central nervous system (CNS) diseases. Loss of NAA has been interpreted as indicating either loss of neurons, or loss of neuron viability. In this investigation, the upregulation of NAA in early stages of construction of the CNS, and its downregulation in experimentally induced damage models of the CNS is reported. The results of this study indicate that the buildup of NAA is not required for viability of neurons in monocellular cultures, and that NAA is lost from multicellular cultured brain slice explants that contain viable neurons. Thus, loss of NAA does not necessarily indicate either loss of neurons or their function. The NAA system, when present in the brain, appears to reflect a high degree of cellular integration, and therefore may be a unique metabolic construct of the intact vertebrate brain. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12728192&query_hl=1 ER - TY - JFULL T1 - . Effect of nitric oxide donor nitroglycerin on bone mineral density in rat model of estrogen deficiency-induced osteopenia A1 - Hukkanen M A1 - Platts LA A1 - Lawes T A1 - Girgis SI A1 - Konttinen YT A1 - Goodship AE A1 - MacIntyre I A1 - Polak JM J1 - Bone Y1 - 2003/// IS - 32 SP - 142 EP - 149 ER - TY - JFULL T1 - In vivo imaging of transcriptionally active estrogen receptors. A1 - Ciana, P A1 - Raviscioni, M A1 - Mussi, P A1 - Vegeto, E A1 - Que, I A1 - Parker, MG A1 - Lowik, C A1 - Maggi, A J1 - Nat Med Y1 - 2003/01// VL - 9 SN - 1078-8956 SP - 82 EP - 86 N2 - Through intracellular receptors, estrogens control growth, differentiation and function of not only reproductive tissues, but also other systems. Estrogen receptors are ligand-dependent transcription factors whose activity is modulated either by estrogens, or by alternative intracellular signaling pathways downstream of growth factors and neurotransmitters. To determine the dynamics of estrogen receptor activity and the dependence of estrogen receptor on 17beta-estradiol in vivo, we generated a transgenic mouse that expresses a luciferase reporter gene under the control of activated estrogen receptors. As expected, luciferase activity, monitored with a cooled charged coupled device camera, paralleled circulating estrogen levels in reproductive tissues and in liver, indicating that the peak transcriptional activity of the estrogen receptor occurred at proestrus. In contrast, in tissues such as bone and brain, the peak activity of estrogen receptors was observed at diestrus. These tissue-specific responses are masked when mice undergo conventional hormone treatment. We also demonstrate that estrogen receptors are active in immature mice before gonadal production of sex hormones as well as in ovariectomized adult mice. These findings emphasize the importance of hormone-independent activation of the estrogen receptor, and have implications for the therapeutic use of estrogens, such as hormone replacement therapy. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12483206&query_hl=1 ER - TY - JFULL T1 - The effect of 58S bioactive sol-gel derived foams on the growth of murine lung epithelial cells A1 - Tan, A A1 - Romanska, HM A1 - Lenza, R A1 - Jones, J A1 - Hench, LL A1 - Polak, JM A1 - Bishop, AE J1 - KEY ENG MAT Y1 - 2003/// VL - 240-2 SP - 719 EP - 723 N2 - Bioactive 58S sol-gel derived foams were tested for their ability to promote the growth and proliferation of murine lung epithelial cells. Foams were modified with an amine or mercaptan functional group and/or coated with laminin. Routine histologic staining with haematoxylin and eosin (H&E) determined qualitatively that cells do grow into foams and their proliferation was evaluated quantitatively with the WST-1 assay. 58S foams promote the growth of MLE-12 cells with the amine modified, laminin coated foam being the most effective. SEM confirmed the above observations and showed that laminin encouraged the growth, attachment and migration of cells into foams and the type of surface modification affects the pattern of growth of cells on the foams. These results provide a foundation for use of biomaterials as bioactive scaffolds promoting differentiation of stem cells into mature pneumocytes. ER - TY - JFULL T1 - Cigarette smoke decreases inducible nitric oxide synthase in lung epithelial cells A1 - Hoyt JC A1 - Robbins RA A1 - Habib M A1 - Springall DR A1 - Buttery LD A1 - Polak JM A1 - Barnes PJ J1 - Exp.Lung Res Y1 - 2003/// IS - 29 SP - 17 EP - 28 ER - TY - JFULL T1 - Development of novel selective cell ablation in the mammary gland and brain to study cell-cell interactions and chemoprevention. A1 - Gusterson, BA A1 - Cui, W A1 - Clark, AJ J1 - Recent Results Cancer Res Y1 - 2003/// VL - 163 SN - 0080-0015 N2 - We have generated transgenic mice which express the gene encoding Escherichia coli nitroreductase (NTR) specifically in the luminal epithelial cells of the mammary gland and the glial cells of the brain. The enzyme activates an antitumour drug CB 1954, to produce a cross-linking agent that kills all cells expressing the enzyme. We have shown that administration of the antitumour drug CB 1954 rapidly and selectively kills these cells. Original experiments demonstrated the ability to ablate the luminal cells in the mammary gland with no apparent bystander effect. Subsequently, astrocytes expressing nitroreductase under the targeting of the GFAP promoter were selectively ablated following administration of the prodrug CB 1954 produces a degeneration of granular neurones due to changes in glutamate levels. Recent experiments demonstrated inhibition of myc-dependent mammary tumours using the same enzyme (nitroreductase)-prodrug (CB 1954), combination. Owing to the ease of control of NTR-mediated cell ablation, we anticipate that this system will supersede herpes simplex virus type 1 thymidine kinase. There are widespread potential applications for this approach in the dissection of complex cellular interactions during development and in the adult organism. The present transgenic models also have important applications for the study in vivo of novel prodrugs that can be selected for variable degrees of bystander effects. Such studies will have particular significance for those groups advocating the use of NTR as an appropriate enzyme for gene-directed enzyme prodrug therapy by providing models of a wide range of human disease for mechanistic and therapeutic experimentation. The results clearly demonstrate that the model has potential to study chemoprevention and fundamental questions on cell-cell interactions in cell biology. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12903841&query_hl=1 ER - TY - JFULL T1 - Unfriendly chemicals in pancreatic cancer. A1 - Khan, SA A1 - Beck, A A1 - Carmichael, PL A1 - Taylor-Robinson, SD J1 - Pancreatology Y1 - 2003/// VL - 3 SN - 1424-3903 SP - 7 EP - 8 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12683401&query_hl=1 ER - TY - JFULL T1 - The effect of 58S bioactive gel-glass sol-gel derived foams on the growth of murine lung epithelial cells A1 - Tan A A1 - Romanska HM A1 - Lenza R A1 - Jones JR A1 - Hench LH A1 - Polak JM A1 - Bishop AE J1 - Key Engineering Materials Y1 - 2003/// SP - 719 EP - 724 ER - TY - JFULL T1 - Fingerprinting the circulating repertoire of antibodies from cancer patients A1 - Mintz, PJ A1 - Kim, J A1 - Do, KA A1 - Wang, XM A1 - Zinner, RG A1 - Cristofanilli, M A1 - Arap, MA A1 - Hong, WK A1 - Troncoso, P A1 - Logothetis, CJ A1 - Pasqualini, R A1 - Arap, W J1 - Nat Biotechnol Y1 - 2003/01// IS - 1 VL - 21 SN - 1087-0156 SP - 57 EP - 63 ER - TY - JFULL T1 - Application of FTIR and Raman spectroscopy to characterisation of bioactive materials and living cells A1 - Notingher, I A1 - Jones, JR A1 - Verrier, S A1 - Bisson, I A1 - Embanga, P A1 - Edwards, P A1 - Polak, JM A1 - Hench, LL J1 - SPECTROSC-INT J Y1 - 2003/// VL - 17 SN - 0712-4813 SP - 275 EP - 288 N2 - Both Fourier Transform Infrared (FTIR) and Raman spectroscopy have been applied to the in vitro characterisation of biomaterials, mainly surface reactions leading to the formation of a biologically active hydroxycarbonate apatite (HCA) layer on the sample surface when immersed in simulated body fluids (SBF). The HCA layer indicates the degree of bioactivity of the sample, because it leads to a strong bond between the biomaterial and living tissue. Reflection measurements using FTIR allow quick, non-destructive detection of the HCA layer for solid and powder samples. Due to the low Raman scattering efficiency and low absorption of water in the visible-near infrared region, Raman micro-spectroscopy was successfully used for the in situ characterisation of 20 and 40 mum diameter 45S5 Bioglass(R) fibres. The in situ capabilities of the Raman micro-spectrometer have also been extended to the characterisation of living cells attached on bioinert silica and bioactive 45S5 Bioglass(R) and 58S substrates. Using a high power 785 nm laser, living cells in physiological conditions can be real-time sampled over long periods of time without inducing cell damage and with good signal strength. Cell death can be monitored because it proved to induce strong changes in the Raman signature in the spectral regions 1000-1150 cm(-1) and 1550-1650 cm(-1). ER - TY - JFULL T1 - Induction of inducible nitric oxide synthase, argininosuccinate synthase, and GP cyclohydrolase I in arthritic joints of human tumor necrosis factor-a transgenic mice A1 - Hukkanen M A1 - Platts LA A1 - Haralambous S A1 - Ainola M A1 - Konttinen YT A1 - Kollias G A1 - Polak JM J1 - J Rheumatol Y1 - 2003/// IS - 30 SP - 652 EP - 659 ER - TY - JFULL T1 - Evidence for protein phosphatase inhibitor-1 playing an amplifier role in beta-adrenergic signaling in cardiac myocytes A1 - El-Armouche, A A1 - Rau, T A1 - Zolk, O A1 - Ditz, D A1 - Pamminger, T A1 - Zimmermann, WH A1 - Jackel, E A1 - Harding, SE A1 - Boknik, P A1 - Neumann, J A1 - Eschenhagen, T J1 - FASEB J Y1 - 2003/01// VL - 17 SN - 0892-6638 SP - 437 EP - + N2 - The protein phosphatase inhibitor-1 (PPI-1) inhibits phosphatase type-1 (PP1) only when phosphorylated by protein kinase A and could play a pivotal role in the phosphorylation/dephosphorylation balance. Rat cardiac PPI-1 was cloned by reverse transcriptase-polymerase chain reaction, expressed in Eschericia coli, evaluated in phosphatase assays, and used to generate an antiserum. An adenovirus was constructed encoding PPI-1 and green fluorescent protein (GFP) under separate cytomegalovirus promotors (AdPPI-1/GFP). A GFP-only virus (AdGFP) served as control. Engineered heart tissue (EHT) from neonatal rat cardiomyocytes and adult rat cardiac myocytes (ARCMs) were used as model systems. PPI-1 expression was determined in human ventricular samples by Northern blots. Compared with AdGFP, AdPPI-1/GFP-infected neonatal rat cardiomyocytes displayed a 73% reduction in PP1 activity. EHTs infected with AdPPI-1/GFP exhibited a fivefold increase in isoprenaline sensitivity. AdPPI-1/GFP-infected ARCMs displayed enhanced cell shortening as well as enhanced phospholamban phosphorylation when stimulated with 1 nM isoprenaline. PPI-1 mRNA levels were reduced by 57+/-12% in failing hearts with dilated and ischemic cardiomyopathy (n=8 each) compared with nonfailing hearts (n=8). In summary, increased PPI-1 expression enhances myocyte sensitivity to isoprenaline, indicating that PPI-1 acts as an amplifier in beta-adrenergic signaling. Decreased PPI-1 in failing human hearts could participate in desensitization of the cAMP pathway. ER - TY - JFULL T1 - Of mice and men...and elephants. A1 - Gordon, MY A1 - Lewis, JL A1 - Marley, SB J1 - Blood Y1 - 2002/12/15/ VL - 100 SN - 0006-4971 SP - 4679 EP - 4680 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12453884&query_hl=1 ER - TY - JFULL T1 - A role for the Fas/Fas ligand apoptotic pathway in regulating myeloid progenitor cell kinetics. A1 - Alenzi, FQ A1 - Marley, SB A1 - Lewis, JL A1 - Chandrashekran, A A1 - Warrens, AN A1 - Goldman, JM A1 - Gordon, MY J1 - Exp Hematol Y1 - 2002/12// VL - 30 SN - 0301-472X SP - 1428 EP - 1435 N2 - Bone marrow from wild-type mice and mice with mutated Fas (lpr) or mutated Fas ligand (gld) was used to investigate the role of the Fas/FasL system in the regulation of myeloid progenitor cell kinetics.Granulocyte-macrophage colony-forming cells (CFU-GM) were measured by a standard colony assay and the proliferative activity of CFU-GM was measured by replating primary colonies and observing secondary colony formation. Fas expression was restored to lpr mouse bone marrow cells by retrovirus-mediated gene transfer and gld mouse marrow cells were treated with soluble FasL. Wild-type marrow cells were treated with YVAD (a caspase inhibitor) or anti-Fas monoclonal antibodies.There were greater frequencies of myeloid progenitor cells (CFU-GM) in lpr and gld mouse marrow compared to wild-type (WT) marrow (p = 0.0008). The proliferative capacity of CFU-GM was also significantly greater for lpr and gld CFU-GM compared to WT CFU-GM (p = 0.0003 and 0.0001, respectively). Retrovirus-mediated restoration of Fas into lpr marrow, and provision of soluble FasL (sFasL) to gld CFU-GM reduced CFU-GM proliferation to WT levels. Treatment of WT CFU-GM with YVAD or anti-FasL monoclonal antibody increased CFU-GM proliferation to the levels found in lpr and gld CFU-GM. YVAD significantly increased and anti-Fas significantly reduced the proliferative capacity of human CFU-GM (p = 0.015 and 0.04, respectively).Fas, FasL, and caspase activation may play an important role in regulating myeloid progenitor cell kinetics. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12482505&query_hl=1 ER - TY - JFULL T1 - Adenovirally-overexpressed beta 2-adrenoceptors enhance the contractile response to CGP 12177A in adult rat cardiomyocytes A1 - Lewis, CJ A1 - Gong, H A1 - Koch, WJ A1 - Brown, MJ A1 - Harding, SE J1 - BRIT J PHARMACOL Y1 - 2002/12// VL - 137 SN - 0007-1188 ER - TY - JFULL T1 - Changing international trends in mortality rates for liver, biliary and pancreatic tumours. A1 - Khan, SA A1 - Taylor-Robinson, SD A1 - Toledano, MB A1 - Beck, A A1 - Elliott, P A1 - Thomas, HC J1 - J Hepatol Y1 - 2002/12// VL - 37 SN - 0168-8278 SP - 806 EP - 813 N2 - BACKGROUND/AIMS: The age-standardized mortality rate for hepatocellular carcinoma is increasing in several countries. However, in England and Wales we previously reported an increase in mortality rates from intrahepatic cholangiocarcinoma. Trends in cholangiocarcinoma in most other industrialized countries are unknown. To further study trends in hepatobiliary and pancreatic tumours, we analysed mortality data from the United States, Japan, Australia and Europe. METHODS: Age-standardized mortality rates for men and women for subcategories of liver tumours, tumours of the gall bladder and extrahepatic biliary tree and pancreas from 1979 to 1998 were obtained from the World Health Organization mortality database. RESULTS: We confirmed previously reported increases in hepatocellular carcinoma, but also found increases in other countries, particularly Australia (3-year average rise from 1.20 to 2.27, men). Mortality for intrahepatic cholangiocarcinoma increased in men in all countries studied, with the largest increases in Australia (from 0.10 to 0.70) and England and Wales (from 0.20 to 0.83). CONCLUSIONS: We present a hitherto unreported rise in age-standardized mortality rates from intrahepatic cholangiocarcinoma across four continents. The cause remains uncertain. An impact on the observed trends of improved diagnostic techniques and death certificate misclassification cannot be completely ruled out. Future research should include epidemiological studies to examine possible case-clustering and investigation of potential aetiological and host factors. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12445422&query_hl=1 ER - TY - JFULL T1 - Hypoxic induction of cox-2 regulates proliferation of human pulmonary artery smooth muscle cells. A1 - Yang, X A1 - Sheares, KK A1 - Davie, N A1 - Upton, PD A1 - Taylor, GW A1 - Horsley, J A1 - Wharton, J A1 - Morrell, NW J1 - Am J Respir Cell Mol Biol Y1 - 2002/12// VL - 27 SN - 1044-1549 SP - 688 EP - 696 N2 - Chronic hypoxia-induced pulmonary hypertension results partly from proliferation of smooth muscle cells in small peripheral pulmonary arteries. Therefore, we examined the effect of hypoxia on growth of pulmonary artery smooth muscle cells (PASMCs) from human distal pulmonary arteries. Initial studies identified that serum-induced proliferation of explant-derived PASMCs was inhibited under hypoxic conditions (3-4 kPa in medium). However, selection of hypoxia-stimulated cells was achieved by culturing cells at low density under conditions of prolonged hypoxia (1-2 wk). In hypoxia-inhibited and -stimulated cells, Western blotting revealed hypoxic induction of cyclooxygenase (COX)-2, which was dependent on the activation of p38(MAPK), but not COX-1, inducible nitric oxide synthase (iNOS), or hemoxygenase-1 (HO-1). Hypoxic induction of COX-2 was also observed in the media of pulmonary arteries in lung organ culture. Hypoxia induced a 4- to 5-fold increase (P < 0.001) in prostaglandin (PG)E(2), PGD(2), PGF(2alpha), and 6-keto-PGF(1alpha) release from PASMCs. Hypoxic inhibition of proliferation was attenuated by incubation with indomethacin (10 micro M), or the COX-2 antagonist, NS398 (10 micro M), but not by the COX-1 antagonist, valeryl salicylate (0.5 mM). In conclusion, we have isolated cells from human peripheral pulmonary arteries that are either inhibited or stimulated by culture under hypoxic conditions. In both cell types hypoxia modulates cell proliferation by induction of COX-2 and production of antiproliferative prostaglandins. Induction of COX-2 may contribute to the inhibition of hypoxia-induced pulmonary vascular remodeling. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12444028&query_hl=1 ER - TY - JFULL T1 - Dissociation of hypertrophic growth from changes in myocyte contractile function. A1 - Harding, SE A1 - Del Monte, F J1 - J Card Fail Y1 - 2002/12// VL - 8 SN - 1071-9164 SP - S415 EP - S420 N2 - Increases in myocyte size and impairment of contractile function are both features of the pathophysiologic hypertrophic process. In this short review, we re-present evidence, both in human disease and in animal models, that the 2 phenomena can be dissociated in both time and relative degree. Additionally, in an animal model, physiologic growth concurrent with pathophysiologic hypertrophy induces similar changes in myocyte macroarchitecture but without the deleterious consequences for contractile function. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12555154&query_hl=1 ER - TY - JFULL T1 - Ion channels in small cells and subcellular structures can be studied with a smart patch-clamp system. A1 - Gorelik, J A1 - Gu, Y A1 - Spohr, HA A1 - Shevchuk, AI A1 - Lab, MJ A1 - Harding, SE A1 - Edwards, CR A1 - Whitaker, M A1 - Moss, GW A1 - Benton, DC A1 - Sánchez, D A1 - Darszon, A A1 - Vodyanoy, I A1 - Klenerman, D A1 - Korchev, YE J1 - Biophys J Y1 - 2002/12// VL - 83 SN - 0006-3495 SP - 3296 EP - 3303 N2 - We have developed a scanning patch-clamp technique that facilitates single-channel recording from small cells and submicron cellular structures that are inaccessible by conventional methods. The scanning patch-clamp technique combines scanning ion conductance microscopy and patch-clamp recording through a single glass nanopipette probe. In this method the nanopipette is first scanned over a cell surface, using current feedback, to obtain a high-resolution topographic image. This same pipette is then used to make the patch-clamp recording. Because image information is obtained via the patch electrode it can be used to position the pipette onto a cell with nanometer precision. The utility of this technique is demonstrated by obtaining ion channel recordings from the top of epithelial microvilli and openings of cardiomyocyte T-tubules. Furthermore, for the first time we have demonstrated that it is possible to record ion channels from very small cells, such as sperm cells, under physiological conditions as well as record from cellular microstructures such as submicron neuronal processes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12496097&query_hl=1 ER - TY - JFULL T1 - Interaction between increased SERCA2a activity and beta -adrenoceptor stimulation in adult rabbit myocytes. A1 - Chaudhri, B A1 - Del Monte, F A1 - Hajjar, RJ A1 - Harding, SE J1 - Am J Physiol Heart Circ Physiol Y1 - 2002/12// VL - 283 SN - 0363-6135 SP - H2450 EP - H2457 N2 - Sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA)2a overexpression and phospholamban depletion have been shown to have beneficial effects on contractility in heart failure. However, the high sympathetic tone during development of failure may interact with increases in SERCA2a activity in potentially deleterious ways. We used adenoviral vectors to overexpress SERCA2a or partially downregulate phospholamban in adult rabbit ventricular myocytes in culture and studied the responses of these cells to beta-adrenoceptor stimulation. SERCA2a overexpression and phospholamban depletion had quantitatively similar effects on basal contraction amplitude and in accelerating relaxation. Increasing SERCA2a activity by either strategy had little effect on the increase in contraction amplitude or incidence of arrhythmias with increasing isoproterenol. Maximum acceleration of relaxation by beta-adrenoceptor stimulation was similar to that produced by SERCA2a overexpression. Isoproterenol treatment of SERCA2a-overexpressing or phospholamban-deficient myocytes produced a further modest decrease in relaxation time, with similar final values in both groups. We find no evidence for Ca(2+) overload induced by SERCA2a overexpression alone or in combination with catecholamines. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12388307&query_hl=1 ER - TY - JFULL T1 - Delayed hypothermia prevents decreases in N-acetylaspartate and reduced glutathione in the cerebral cortex of the neonatal pig following transient hypoxia-ischaemia. A1 - Brooks, KJ A1 - Hargreaves, I A1 - Bhakoo, K A1 - Sellwood, M A1 - O'Brien, F A1 - Noone, M A1 - Sakata, Y A1 - Cady, E A1 - Wylezinska, M A1 - Thornton, J A1 - Ordidge, R A1 - Nguyen, Q A1 - Clemence, M A1 - Wyatt, J A1 - Bates, TE J1 - Neurochem Res Y1 - 2002/12// VL - 27 SN - 0364-3190 SP - 1599 EP - 1604 N2 - The effects of normothermia and delayed hypothermia on the levels of N-acetylaspartate (NAA), reduced glutathione (GSH) and the activities of mitochondrial complex I, II-III, IV and citrate synthase were measured in brain homogenates obtained from anaesthetized neonatal pigs following transient in vivo hypoxia-ischaemia. In the normothermic animals there was a significant decrease in complex I activity and in the levels of GSH and NAA when compared to the controls. Delayed hypothermia preserved NAA and GSH at control levels and enhanced the rate of complex II-III activity. There was correlation (R = 0.79) between GSH and NAA levels when data from all three experimental groups were analyzed. Citrate synthase activity was not significantly different in the three groups, indicating maintenance of mitochondrial integrity. These data suggest that delayed hypothermia affords protection of integrated mitochondrial function in the neonatal brain following transient hypoxia-ischaemia. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12515311&query_hl=1 ER - TY - JFULL T1 - Autologous stem cell transplantation improves abnormal bone turnover in multiple myeloma. A1 - Terpos, E A1 - Hatziharissi, E A1 - Szydlo, R A1 - de la Fuente, J A1 - Karadimitris, A A1 - Kanfer, E A1 - Olavarria, E A1 - Goldman, JM A1 - Apperley, JF A1 - Rahemtulla, A J1 - BLOOD Y1 - 2002/11/16/ VL - 100 SN - 0006-4971 SP - 434A EP - 434A ER - TY - JFULL T1 - Hemocytopenia related to abnormal immunity detected by a modified coombs test for measuring autoantibody of bone marrow mononuclear cells. A1 - He, H A1 - Shao, ZH A1 - Fu, R A1 - Liu, H A1 - Cao, Z A1 - Tian, P A1 - Song, LY A1 - Sun, J A1 - Lia, HR A1 - Cui, W A1 - Chu, YL A1 - Qian, LS A1 - Yang, TY J1 - BLOOD Y1 - 2002/11/16/ VL - 100 SN - 0006-4971 SP - 136B EP - 136B ER - TY - JFULL T1 - Risk stratification using cardiac troponin T for adverse outcomes within 3 years in 395 outpatients on chronic hemodialysis A1 - Ishii, J A1 - Toriyama, T A1 - Nomura, M A1 - Cui, W A1 - Nakamura, Y A1 - Naruse, H A1 - Mori, Y A1 - Kumada, Y A1 - Takahashi, H A1 - Kawahara, H A1 - Hishida, H J1 - CIRCULATION Y1 - 2002/11/05/ VL - 106 SN - 0009-7322 SP - 347 EP - 347 ER - TY - JFULL T1 - Increased serum concentration of oxidized lipoprotein (a) is a new risk factor for coronary artery disease A1 - Ishii, J A1 - Nomura, M A1 - Cui, W A1 - Nakamura, Y A1 - Naruse, H A1 - Mori, Y A1 - Hishida, H A1 - Yamada, S J1 - CIRCULATION Y1 - 2002/11/05/ VL - 106 SN - 0009-7322 SP - 347 EP - 347 ER - TY - JFULL T1 - Meiotic chromosome segregation in Drosophila A1 - Hawley, R A1 - Harris, DT A1 - Cui, W A1 - Kramer, JJ A1 - Page, SL J1 - MOL BIOL CELL Y1 - 2002/11// VL - 13 SN - 1059-1524 SP - 150A EP - 150A ER - TY - JFULL T1 - Enhanced DNA-directed effects of FdUMP[10] compared to 5-FU A1 - Gmeiner, W A1 - Liu, J A1 - Cui, W A1 - Willingham, M J1 - EUR J CANCER Y1 - 2002/11// VL - 38 SN - 0959-8049 SP - S23 EP - S24 ER - TY - JFULL T1 - Guidelines for the diagnosis and treatment of cholangiocarcinoma: consensus document A1 - Khan, SA A1 - Davidson, BR A1 - Goldin, R A1 - Pereira, SP A1 - Rosenberg, WMC A1 - Taylor-Robinson, SD A1 - Thillainayagam, AV A1 - Thomas, HC A1 - Thursz, MR A1 - Wasan, H J1 - GUT Y1 - 2002/11// VL - 51 SN - 0017-5749 SP - 1 EP - 9 ER - TY - JFULL T1 - Functional consequences of Na/Ca exchanger overexpression in cardiac myocytes. A1 - Terracciano, C J1 - Ann N Y Acad Sci Y1 - 2002/11// VL - 976 SN - 0077-8923 SP - 520 EP - 527 N2 - Several factors are important in the relationship between Na/Ca exchanger overexpression and Ca cycling in physiological and pathophysiological situations. First, there are species differences. Transgenic mouse cardiac myocytes overexpressing Na/Ca exchanger showed a faster Ca transient associated with increased sarcoplasmic reticulum (SR) Ca content compared with wild-type myocytes. Cultured rabbit cardiac myocytes overexpressing Na/Ca exchanger showed reduced amplitude of the Ca transient and reduced SR Ca content. Second, the activity of other Ca regulatory proteins have to be considered. When Ca uptake via SR Ca ATPase (SERCA) was reduced by thapsigargin in transgenic mouse myocytes overexpressing Na/Ca exchanger, the time course of the Ca transient was slowed, and the SR Ca content was reduced to wild-type mouse myocyte levels, suggesting a potential compensatory role of Na/Ca exchanger overexpression when SERCA function is reduced. Finally, there are confounding factors related to the pathophysiological conditions. Our results suggest that Na/Ca exchanger overexpression can compensate for defects in SR Ca uptake in mouse myocytes. The consequences of Na/Ca exchanger overexpression in other species and conditions is unpredictable and require further investigation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12502606&query_hl=1 ER - TY - JFULL T1 - Guidelines for the diagnosis and treatment of cholangiocarcinoma: consensus document. A1 - Khan, SA A1 - Davidson, BR A1 - Goldin, R A1 - Pereira, SP A1 - Rosenberg, WM A1 - Taylor-Robinson, SD A1 - Thillainayagam, AV A1 - Thomas, HC A1 - Thursz, MR A1 - Wasan, H A1 - British Society of Gastroenterology J1 - Gut Y1 - 2002/11// VL - 51 Suppl 6 SN - 0017-5749 SP - VI1 EP - VI9 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12376491&query_hl=1 ER - TY - JFULL T1 - Inducible ablation of adipocytes in adult transgenic mice expressing the E. coli nitroreductase gene. A1 - Felmer, R A1 - Cui, W A1 - Clark, AJ J1 - J Endocrinol Y1 - 2002/11// VL - 175 SN - 0022-0795 SP - 487 EP - 498 N2 - We describe the use of an enzyme prodrug system based on E. coli nitroreductase (NTR) to achieve the specific ablation of adipose tissue. Transgenic mice expressing the NTR gene specifically in the adipose tissue were generated using the adipocyte specific promoter aP2. After treatment with the prodrug CB1954 these mice showed extensive cell depletion in all fat depots; this was directly correlated to both the dose of prodrug and the levels of NTR expression. Higher doses of CB1954 resulted in complete disappearance of visible adipose stores in some transgenic mice. These mice exhibited an impaired ability to thermoregulate body temperature. Lower doses of CB1954 resulted in a partial reduction of the adipose tissue leaving non-expressing cells that escape ablation. These animals show normal levels of blood glucose and triglycerides but have reduced leptin levels. After 30 days they were able to regenerate the fat depots and leptin levels returned to normal but, interestingly, no NTR-expressing cells were detectable. The present model provides a new approach to manipulate the number of adipocytes at different stages of mouse development and provides a new system for the study of fat metabolism especially in abnormal conditions such as obesity and its modulation through manipulation of the target cell population. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12429046&query_hl=1 ER - TY - JFULL T1 - Stabilization of telomere length and karyotypic stability are directly correlated with the level of hTERT gene expression in primary fibroblasts. A1 - Cui, W A1 - Aslam, S A1 - Fletcher, J A1 - Wylie, D A1 - Clinton, M A1 - Clark, AJ J1 - J Biol Chem Y1 - 2002/10/11/ VL - 277 SN - 0021-9258 SP - 38531 EP - 38539 N2 - Telomere shortening and lack of telomerase activity have been implicated in cellular senescence in human fibroblasts. Expression of the human telomerase (hTERT) gene in sheep fibroblasts reconstitutes telomerase activity and extends their lifespan. However, telomere length is not maintained in all cell lines, even though in vitro telomerase activity is restored in all of them. Cell lines expressing higher levels of hTERT mRNA do not exhibit telomere erosion or genomic instability. By contrast, fibroblasts expressing lower levels of hTERT do exhibit telomere shortening, although the telomeres eventually stabilize at a shorter length. The shorter telomere lengths and the extent of karyotypic abnormalities are both functions of hTERT expression level. We conclude that telomerase activity is required to bypass senescence but is not sufficient to prevent telomere erosion and genomic instability at lower levels of expression. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12122013&query_hl=1 ER - TY - JFULL T1 - Characterization of a myocardial depressant factor in meningococcal septicemia. A1 - Pathan, N A1 - Sandiford, C A1 - Harding, SE A1 - Levin, M J1 - Crit Care Med Y1 - 2002/10// VL - 30 SN - 0090-3493 SP - 2191 EP - 2198 N2 - OBJECTIVE: Identification and characterization of myocardial depressant factors present in meningococcal septicemia. DESIGN: Laboratory investigation of myocardial depression that used isolated cardiac myocytes as an model of cardiac contractile function. SETTING: University hospital and laboratories. PATIENTS: Children with severe meningococcal septic shock requiring intensive care. ANIMALS: Myocytes obtained from adult male Sprague-Dawley rats. INTERVENTIONS: Serum samples obtained from the acute phase of sepsis were evaluated for the presence of myocardial depressant activity. Further characterization of the myocardial depressant factor was undertaken by using cell culture supernatants from whole blood and peripheral blood mononuclear cells that had been exposed to heat-killed meningococci. MEASUREMENTS AND MAIN RESULTS: Myocardial depressant activity was measured by using isolated rat left-ventricular myocytes. Changes in amplitude of contraction and in the speed of contraction and relaxation were determined after cells were exposed to various stimuli. Serum from patients with meningococcal disease had myocardial depressant activity. This activity was also present in whole blood and peripheral blood mononuclear cells exposed to meningococci. Myocardial depressant activity was found to be heat stable, proteinaceous, and of a molecular weight range of 10-25 kDa. The activity did not elevate concentrations of cyclic guanylic acid. Lipopolysaccharide-binding protein augmented the release of myocardial depressant factor by peripheral blood mononuclear cells exposed to meningococci. CONCLUSIONS: Myocardial depression in meningococcal sepsis is mediated in part by circulating myocardial depressant factors. Myocardial depressant factors are also released when whole blood or peripheral blood mononuclear cells of healthy donors are exposed to heat-killed meningococci. Release of the factors appears to be mediated through endotoxin-induced activation of peripheral blood mononuclear cells, since lipopolysaccharide-binding protein augments release in a dose-responsive manner. Partial physicochemical characterization of the factors has been achieved. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12394943&query_hl=1 ER - TY - JFULL T1 - Gene expression: a review of clinical applications in otorhinolaryngology-head and neck surgery. A1 - Vats, A A1 - Tolley, NS A1 - Polak, JM A1 - Knight, BC J1 - Clin Otolaryngol Allied Sci Y1 - 2002/10// VL - 27 SN - 0307-7772 SP - 291 EP - 295 N2 - Tissue engineering is a multidisciplinary area of research aimed at regeneration of tissues and restoration of function of organs through implantation of cells/tissues grown outside the body or stimulating cells to grow into implanted matrix. In this short review, we aim to examine current techniques in gene expression analysis and their relevant clinical applications to the field of otorhinolaryngology-head and neck surgery. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12383283&query_hl=1 ER - TY - JFULL T1 - Spectral profiles of cultured neuronal and glial cells derived from HRMAS (1)H NMR spectroscopy. A1 - Griffin, JL A1 - Bollard, M A1 - Nicholson, JK A1 - Bhakoo, K J1 - NMR Biomed Y1 - 2002/10// VL - 15 SN - 0952-3480 SP - 375 EP - 384 N2 - In the investigations of brain function and pathology in vivo by magnetic resonance spectroscopy (MRS), a decrease in the relative concentration of N-acetyl aspartate (NAA) has been correlated with neuronal cell damage or loss, while a relative increase in the resonance intensity of creatine has been correlated with gliosis. However, neither metabolite is confined strictly to one cell-type. In this study, pattern recognition of spectra derived from high-resolution magic angle spinning (HRMAS) (1)H NMR spectroscopy was used to distinguish three neural cell types; cortical astrocytes, cerebellar neurones and O-2A progenitors. The intact cells contained significant amounts of lipid resonances (-CH(2)CH(3) and -CH(2)CH(2)CH(2)-) in all three cell-types, even when a T(2)-edited Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence was used, selectively attenuating resonances from macromolecules. Creatine was also detected in all three cell types. Principle component analysis (PCA) readily differentiated the NMR spectra, based on the individual metabolic profile derived from the cohort of cell type examined using conventional solvent-suppressed and CPMG pulse sequences. Creatine was not found to contribute to this separation. Moreover, the large lipid content of neuronal cells contributed most to the separation from the other cell types. This suggests that during MRS in vivo, where lipid resonances are commonly 'edited out' by T(2) delays, significant information may be sacrificed concerning relative contribution from individual cell types. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12357551&query_hl=1 ER - TY - JFULL T1 - Modulation of hydrogen peroxide induced injury to corneal endothelium by virus mediated catalase gene transfer. A1 - Hudde, T A1 - Comer, RM A1 - Kinsella, MT A1 - Buttery, L A1 - Luthert, PJ A1 - Polak, JM A1 - George, AJ A1 - Larkin, DF J1 - Br J Ophthalmol Y1 - 2002/09// VL - 86 SN - 0007-1161 SP - 1058 EP - 1062 N2 - AIM: To examine the effect of catalase gene transfer on survival of corneal endothelial cells (EC) following challenge with hydrogen peroxide (H(2)O(2)) in an ex vivo model of oxidative stress. METHODS: A recombinant adenovirus vector (AdCL) was used to transfer human catalase cDNA into EC of whole thickness rabbit corneas ex vivo. The resulting catalase protein concentration was measured in corneal lysates by ELISA; catalase functional activity in lysates was determined using a H(2)O(2) activity assay. To examine the morphological effects of catalase gene transfer in modulation of H(2)O(2) induced injury, transduced corneas were maintained in ex vivo culture and challenged with H(2)O(2). Laser scanning confocal microscopy was used to image EC injury. Cell density, cell morphology, and ratios of viable to necrotic cells were determined. RESULTS: Following incubation with AdCL, catalase expression reached maximum at 5-7 days. Corneas transduced with AdCL showed increased EC cell survival following challenge with H(2)O(2) on day 3 when compared to null vector control or mock infected corneas. CONCLUSIONS: Ex vivo catalase gene transfer can protect EC from death mediated by H(2)O(2). This gene based approach to the protection of corneal endothelium from oxidative stress may have application in prevention of EC loss in pathological conditions in which H(2)O(2) is involved and in ex vivo donor corneal storage before transplantation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12185138&query_hl=1 ER - TY - JFULL T1 - Immunocytochemical evidence for inducible nitric oxide synthase and cyclooxygenase-2 expression with nitrotyrosine formation in human hibernating myocardium. A1 - Baker, CS A1 - Dutka, DP A1 - Pagano, D A1 - Rimoldi, O A1 - Pitt, M A1 - Hall, RJ A1 - Polak, JM A1 - Bonser, RS A1 - Camici, PG J1 - Basic Res Cardiol Y1 - 2002/09// VL - 97 SN - 0300-8428 SP - 409 EP - 415 N2 - BACKGROUND: Myocardial hibernation may result from repetitive episodes of transient ischaemia leading to prolonged dysfunction. Inducible nitric oxide synthase (iNOS) expression has been demonstrated in animals following brief, non-lethal ischaemia-reperfusion injury. We therefore, hypothesised that in human hibernating myocardium: 1). iNOS would be present; 2). the reaction of nitric oxide and superoxide would form the strong oxidant peroxynitrite; 3) that this process would be accompanied by the expression of cyclooxygenase-2 (Cox-2) which interacts with NOS and whose products could further affect myocardial function. METHOD AND RESULTS: In sixteen patients with coronary artery disease (CAD), left ventricular biopsies were obtained from chronically dysfunctional segments subtended by a stenotic artery (> 75 %) and shown to be viable by (18)F-fluorodeoxyglucose positron emission tomography. Comparison was made with myocardial biopsies (n = 8) from normally contracting myocardium in patients undergoing coronary surgery, from unused transplant donors and at post-mortem. Regional wall motion score improved in all patients 6 months post-revascularisation (from 2.7 +/- 0.7 to 1.5 +/- 0.5; p < 0.001), confirming hibernation. Immunocytochemistry localized reactivity to iNOS, Cox-2 and nitrotyrosine (a marker of peroxynitrite formation) to cardiomyocytes from hibernating segments. No difference in reactivity to endothelial NOS was seen between hibernating and control cardiomyocytes. CONCLUSION: Cox-2 and iNOS are co-expressed in hibernating myocardium with nitrotyrosine suggesting nitric oxide production and peroxynitrite formation. We propose that this is secondary to ischaemia-reperfusion and that the products of these enzymes may have consequences for myocardial contractile function and survival. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12200641&query_hl=1 ER - TY - JFULL T1 - Hierarchy of carbon source selection in Paracoccus pantotrophus: strict correlation between reduction state of the carbon substrate and aerobic expression of the nap operon. A1 - Ellington, MJ A1 - Bhakoo, KK A1 - Sawers, G A1 - Richardson, DJ A1 - Ferguson, SJ J1 - J Bacteriol Y1 - 2002/09// VL - 184 SN - 0021-9193 SP - 4767 EP - 4774 N2 - Paracoccus pantotrophus can express a periplasmic nitrate reductase (Nap) during aerobic growth. A proposed role for this enzyme is the dissipation of excess redox energy during oxidative metabolism of reduced carbon substrates. To investigate the regulation of nap expression, a transcriptional fusion between the nap promoter region of P. pantotrophus and the lacZ gene was constructed. When this fusion was used, analyses showed that transcription from the nap promoter increases as the average reduction state of the carbon atoms increases. Thus, beta-galactosidase activities increase as the carbon source changes in the order succinate-acetate-butyrate. This result was obtained regardless of which of the three carbon sources was used for culture of the inoculum. If two carbon sources were presented together, the beta-galactosidase activity was always the same as it was when the least-reduced carbon source was added alone. This suggests that the regulation is dependent upon metabolism of the more-reduced carbon sources rather than just their presence in the medium. Analysis of culture medium by (1)H nuclear magnetic resonance showed that for aerobic growth P. pantotrophus strictly selected its carbon source in the order succinate-acetate-butyrate. This was reflected by diauxic growth kinetics on medium containing mixed carbon substrates. The regulatory mechanism underpinning such a selection is unknown but is likely to be related to the mechanism which controls the transcription of the nap operon. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12169601&query_hl=1 ER - TY - JFULL T1 - Taurocholate induces changes in rat cardiomyocyte contraction and calcium dynamics. A1 - Gorelik, J A1 - Harding, SE A1 - Shevchuk, AI A1 - Koralage, D A1 - Lab, M A1 - de Swiet, M A1 - Korchev, Y A1 - Williamson, C J1 - Clin Sci (Lond) Y1 - 2002/08// VL - 103 SN - 0143-5221 SP - 191 EP - 200 N2 - Obstetric cholestasis is characterized by raised bile acids, and can be complicated by intrauterine death. We have shown that the bile acid taurocholate causes loss of synchronous beating, bradycardia and cessation of contraction in cultured rat cardiomyocytes [Williamson, Gorelik, Eaton, Lab, de Swiet and Korchev (2001) Clin. Sci. 100, 363-369]. The aim of the present study was to investigate the effect of taurocholate on cardiomyocytes further. We demonstrated a reduced rate of contraction and proportion of beating cells when rat cardiomyocytes were exposed to increasing concentrations of taurocholate (0.1-3.0 mM); more marked at higher concentrations (P<0.001). Using scanning ion-conductance microscopy, we also demonstrated reduced amplitude of contraction and calcium transients with taurocholate. Our observations indicate that taurocholate affects calcium release from the sarcoplasmic reticulum and this parallels changes in contractile function. The relationship between the contraction amplitude and calcium transient is not linear, particularly at higher concentrations of taurocholate. We observed different effects in individual cultured neonatal cells; a reversible reduction in rate and amplitude of contraction in some, and irreversible oscillatory (fibrillatory) cessation of beating in others. The effects were more marked with higher concentrations. The contraction amplitude was also reduced in adult cardiomyocytes. The changes were reversible following removal of taurocholate in adult, but not in neonatal, cardiomyocytes exposed to higher concentrations (>0.3 mM) (P<0.001). In conclusion we have demonstrated that the bile acid taurocholate can cause different types of dysrhythmia in individual cardiomyocytes. These results provide further support for the hypothesis that obstetric cholestasis may produce cardiac-related sudden intrauterine death. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12149111&query_hl=1 ER - TY - JFULL T1 - Targeting of SWI/SNF chromatin remodelling complexes to estrogen-responsive genes. A1 - Belandia, B A1 - Orford, RL A1 - Hurst, HC A1 - Parker, MG J1 - EMBO J Y1 - 2002/08/01/ VL - 21 SN - 0261-4189 SP - 4094 EP - 4103 N2 - SWI/SNF complexes are ATP-dependent chromatin remodelling enzymes that have been implicated in the regulation of gene expression in yeast and higher eukaryotes. BRG1, a catalytic subunit in the mammalian SWI/SNF complex, is required for transcriptional activation by the estrogen receptor, but the mechanisms by which the complex is recruited to estrogen target genes are unknown. Here, we have identified an interaction between the estrogen receptor and BAF57, a subunit present only in mammalian SWI/SNF complexes, which is stimulated by estrogen and requires both a functional hormone-binding domain and the DNA-binding region of the receptor. We also found an additional interaction between the p160 family of coactivators and BAF57 and demonstrate that the ability of p160 coactivators to potentiate transcription by the estrogen receptor is dependent on BAF57 in transfected cells. Moreover, chromatin immunoprecipitation assays demonstrated that BAF57 is recruited to the estrogen-responsive promoter, pS2, in a ligand-dependent manner. These results suggest that one of the mechanisms for recruiting SWI/SNF complexes to estrogen target genes is by means of BAF57. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12145209&query_hl=1 ER - TY - JFULL T1 - Stem cells: sources and applications. A1 - Vats, A A1 - Tolley, NS A1 - Polak, JM A1 - Buttery, LD J1 - Clin Otolaryngol Allied Sci Y1 - 2002/08// VL - 27 SN - 0307-7772 SP - 227 EP - 232 N2 - Tissue engineering is a multidisciplinary area of research aimed at regeneration of tissues and restoration of function of organs through implantation of cells/tissues grown outside the body, or stimulating cells to grow into implanted matrix. In this short review, some of the most recent developments in the use of stem cells for tissue repair and regeneration will be discussed. There is no doubt that stem cells derived from adult and embryonic sources hold great therapeutic potential but it is clear that there is still much research required before their use is commonplace. There is much debate over adult versus embryonic stem cells and whether both are required. It is probably too early to disregard one or other of these cell sources. With regard to embryonic stem cells, the major concern relates to the ethics of their creation and the proposed practice of therapeutic cloning. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12169121&query_hl=1 ER - TY - JFULL T1 - Derivation of type II alveolar epithelial cells from murine embryonic stem cells. A1 - Ali, NN A1 - Edgar, AJ A1 - Samadikuchaksaraei, A A1 - Timson, CM A1 - Romanska, HM A1 - Polak, JM A1 - Bishop, AE J1 - Tissue Eng Y1 - 2002/08// VL - 8 SN - 1076-3279 SP - 541 EP - 550 N2 - Embryonic stem (ES) cell pluripotency is being investigated increasingly to obtain specific cell lineages for tissue engineering. However, the possibility that ES cells can give rise to lung tissue has not been tested. We hypothesized that lung epithelial cells (type II pneumocytes) can be derived in vitro from murine ES cells. After withdrawal of leukemia inhibitory factor (LIF) and formation of embryoid bodies in maintenance medium for 10, 20, and 30 days, differentiating ES cells were kept in the same medium or transferred to serum-free small airway growth medium (SAGM) for a further 3 or 14 days of culture. The presence of type II pneumocytes in the resulting mixed cultures was demonstrated by reverse transcriptase-polymerase chain reaction (RT-PCR) of surfactant protein C (SPC) mRNA, immunostaining of SPC, and electron microscopy of osmiophilic lamellar bodies only at 30 days sampling time. SAGM appeared to be more favorable for type II cell formation than ES medium. No SPC transcripts were found in differentiating cells grown under the same conditions without formation of embryoid bodies. These findings could form the basis for the enrichment of ES cell-derived cultures with type II pneumocytes, and provide an in vitro system for investigating mechanisms of lung repair and regeneration. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12201994&query_hl=1 ER - TY - JFULL T1 - Differentiating embryonic stem cells: GAPDH, but neither HPRT nor beta-tubulin is suitable as an internal standard for measuring RNA levels. A1 - Murphy, CL A1 - Polak, JM J1 - Tissue Eng Y1 - 2002/08// VL - 8 SN - 1076-3279 SP - 551 EP - 559 N2 - Embryonic stem (ES) cells are pluripotent cell lines that possess virtually unlimited self-renewal and differentiation capacity. Such characteristics make them potentially an invaluable cell source for diverse tissue-engineering applications. In vitro ES cell differentiation occurs spontaneously in three-dimensional structures termed "embryoid bodies" that mimic postimplantation embryonic tissue. HPRT, beta-tubulin, and GAPDH are commonly used as internal RNA standards in ES cell-derived gene transcription studies so that corrected sample mRNA levels can be obtained for (semi) quantitative gene expression data. However, if reliable data is to be obtained, it is essential that such housekeeping gene expression remains constant, and this has not been demonstrated for differentiating ES cell cultures, which represent a mixed and changing population of cells with time in culture. Therefore, in the present study, we tested the suitability of these housekeeping genes to act as true internal standards for differentiating murine ES cells cultured as embryoid bodies. PCR-amplified gene-specific products were quantified from digital images of ethidium bromide-stained gels using a computer software package. Both HPRT and beta-tubulin mRNA levels varied markedly in spontaneously differentiating and growth factor-supplemented (TGF-beta) ES cell cultures (p < 0.001, ANOVA), while GAPDH expression remained relatively constant (p > 0.2). Our results demonstrate the importance of fully validating housekeeping gene expression in in vitro ES cell gene transcription studies and suggest that GAPDH may be a suitable candidate to act as an internal RNA standard, while both HPRT and beta-tubulin appear to be inappropriate. Finally, we demonstrate enhanced mesodermal differentiation of ES cell-derived cultures by treatment with TGF-beta through significant upregulation of Brachyury T expression, with a concomitant decrease in expression of the undifferentiated ES cell marker Oct-4. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12201995&query_hl=1 ER - TY - JFULL T1 - iNOS gene upregulation is associated with the early proliferative response of human lung fibroblasts to cytokine stimulation. A1 - Romanska, HM A1 - Polak, JM A1 - Coleman, RA A1 - James, RS A1 - Harmer, DW A1 - Allen, JC A1 - Bishop, AE J1 - J Pathol Y1 - 2002/07// VL - 197 SN - 0022-3417 SP - 372 EP - 379 N2 - Increased release of oxidants has been implicated in the pathogenesis of pulmonary fibrosis. Previous work in the rat showed that formation of the early fibrotic lesion is associated with increased expression of inducible nitric oxide synthase (iNOS) in pulmonary fibroblasts. The aim of this study was to test the hypothesis that NO is involved in the activation of pulmonary fibroblasts. The effects of endogenous and exogenous NO on proliferation of human pulmonary fibroblasts were investigated by administration of cytomix or SNAP, respectively. At low concentrations, both treatments increased cell numbers, an effect attenuated by iNOS inhibitor or NO scavenger. Induction of iNOS was confirmed by measurement of nitrate/nitrite production and by immunodetection. Quantitative RT-PCR showed an increase in iNOS mRNA as early as 3 h after stimulation. These results support the hypothesis and show that upregulation of the iNOS gene is an early event in the proliferative response of human lung fibroblasts to inflammatory stimuli. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12115884&query_hl=1 ER - TY - JFULL T1 - Embryonic stem cells. A1 - Bishop, AE A1 - Buttery, LD A1 - Polak, JM J1 - J Pathol Y1 - 2002/07// VL - 197 SN - 0022-3417 SP - 424 EP - 429 N2 - The capacity of embryonic stem cells for virtually unlimited self-renewal and differentiation capacity has opened up the prospect of widespread applications in biomedical research and regenerative medicine. For the latter, the cells provide hope that it will be possible to overcome the problems of donor tissue shortage and also, by making the cells immunocompatible with the recipient, implant rejection. Four years after the first derivation of human pluripotent cell lines from pre-implantation embryos, a great deal has been learnt about their biology and how differentiation can be encouraged towards particular cell lineages. However, considerable research is needed, not least into means to enrich and purify derivative cell lineages, before clinical trials can be considered. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12115859&query_hl=1 ER - TY - JFULL T1 - Nitric oxide in the development of obliterative bronchiolitis in a heterotopic pig model. A1 - Salminen, US A1 - Maasilta, PK A1 - Harjula, AL A1 - Romanska, HM A1 - Bishop, AE A1 - Polak, JM J1 - Transplantation Y1 - 2002/06/15/ VL - 73 SN - 0041-1337 SP - 1724 EP - 1729 N2 - BACKGROUND: Inflammation, epithelial cell injury, and development of fibrosis and airway obliteration are the major histological features of posttransplant obliterative bronchiolitis (OB). The expression of inducible nitric oxide synthase (iNOS) in the damaged epithelium, accompanied by peroxynitrite, suggests that endogenous nitric oxide (NO) mediates the epithelial destruction preceding obliteration. To elucidate the role of NO in this cascade, heterotopic bronchial allografts were studied in pigs. METHODS: Allografts or autografts were harvested serially 3-90 days after transplantation and processed for histology and immunocytochemistry for iNOS, nitrotyrosine, a marker of peroxynitrite formation, and superoxide dismutase (SOD). RESULTS: During initial ischemic damage to the epithelium, iNOS, nitrotyrosine, and SOD were found to be strongly expressed in the epithelium of all implants as well as later, after partial recovery, parallel to onset of epithelial destruction and subsequent airway obliteration in allografts. The levels of expression of iNOS in fibroblasts during the early phase of obliteration paralleled the onset of fibrosis. Constant expression of iNOS and SOD, but not nitrotyrosine, occurred in autografts and allografts with blocked alloimmune response. CONCLUSIONS: These findings suggest that an excessive amount of NO promotes posttransplant obliterative bronchiolitis by destroying airway epithelium and stimulating fibroblast activity. SOD may provide protection by binding reactive molecules and preventing peroxynitrite formation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12084993&query_hl=1 ER - TY - JFULL T1 - Developments in therapeutics for pulmonary arterial hypertension. A1 - Wilkins, MR A1 - Møller, GM A1 - Ren, X A1 - Wharton, J J1 - Minerva Cardioangiol Y1 - 2002/06// VL - 50 SN - 0026-4725 SP - 175 EP - 187 N2 - For many years, the management of pulmonary hypertension has been frustrated by an inadequate understanding of its pathology and limited therapeutic options, but this is changing rapidly. Recently, novel insight into the pathogenesis of primary pulmonary hypertension (PPH) has been provided by the demonstration of mutations in BMPR2 and ALK-1 genes in a significant number of patients with the condition. These genes encode members of the TGF-b receptor superfamily and their integrity is important in the maintenance of normal pulmonary vascular structure and function. At the same time, there has been a major advance in the treatment of the condition due to development of 2 orally active pharmacological agents, bosentan and sildenafil, which demonstrate some selectivity for the pulmonary vasculature. This review examines how the management of PPH and severe pulmonary hypertension in associated diseases has changed and looks at exciting future developments. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12107399&query_hl=1 ER - TY - JFULL T1 - Increased inducible nitric oxide synthase and matrix metalloproteinase MMP-9 in the symptomatic carotid artery plaque A1 - Clayton, GS A1 - Nohadani, MR A1 - Golledge, J A1 - Bishop, AE A1 - Greenhalgh, RM A1 - Davies, AH J1 - BRIT J SURG Y1 - 2002/06// VL - 89 SN - 0007-1323 SP - 99 EP - 99 ER - TY - JFULL T1 - A quantitative study of the neural changes underlying pyloric stenosis in dogs. A1 - Abel, RM A1 - Doré, CJ A1 - Bishop, AE A1 - Facer, P A1 - Polak, JM A1 - Spitz, L J1 - Anat Histol Embryol Y1 - 2002/06// VL - 31 SN - 0340-2096 SP - 139 EP - 143 N2 - This study aimed to quantify the neural changes in congenital pyloric stenosis in dogs and to study the comparative anatomy between this condition in dogs and that in infantile hypertrophic pyloric stenosis. Eight specimens from the pylorus of dogs with pyloric stenosis and six control specimens were examined using conventional histology and immunohistochemistry for a range of neural antigens. The changes in the proportion of nerves immunoreactive for each antigen were quantified and analysed statistically. The morphology of the nerves in the diseased dogs was similar to that in controls. Only vasoactive intestinal peptide was reduced in expression in dogs (median proportion in control dogs 0.57, in diseased dogs 0.17; P = 0.065). This study demonstrates both morphological similarities and significant differences between closely related conditions in dogs, humans and other species. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12479355&query_hl=1 ER - TY - JFULL T1 - Distribution of phosphodiesterase type 5 (PDE5) and effect of its inhibition in chronic hypoxic rat lung A1 - Sebkhi, A A1 - Strange, J A1 - Phillips, SC A1 - Wharton, J A1 - Wilkins, MR J1 - J HYPERTENS Y1 - 2002/06// VL - 20 SN - 0263-6352 SP - S281 EP - S281 ER - TY - JFULL T1 - Overexpression of SERCA2a accelerates repolarisation in rabbit ventricular myocytes. A1 - Terracciano, CM A1 - Hajjar, RJ A1 - Harding, SE J1 - Cell Calcium Y1 - 2002/06// VL - 31 SN - 0143-4160 SP - 299 EP - 305 N2 - Overexpression of the sarcoplasmic reticulum Ca ATPase (SERCA2a) produces positive inotropism and it has been proposed as a promising strategy to counteract defective excitation-contraction coupling in the failing heart. However, the effects of overexpressing SERCA2a on action potential duration (APD), which can affect diastolic parameters in the heart, is unknown. We, therefore, investigated the relationship between SERCA2a overexpression and APD in adult rabbit ventricular myocytes which were cultured for 48 h. Overexpression of SERCA2a was achieved by infection with an adenovirus carrying both SERCA2a and GFP independently driven by CMV promoters, Ad.SERCA2a. Myocytes infected with Ad.GFP only and/or non-infected myocytes were used as controls. Electrophysiological measurements were taken using switch clamping with 15-25 M Omega resistance microelectrodes. In Ad.SERCA2a infected myocytes, APD was significantly reduced compared with both groups of control cells at 0.5 Hz (APD50 (ms) non-infected: 481+/-98, n=12; Ad.GFP: 464+/-85, n=11; Ad.SERCA2a: 285+/-69, n=13 (mean+/-S.E.M.) and at 1 Hz (APD50 (ms) non-infected: 375+/-64, n=22; Ad.GFP: 363+/-47, n=18; Ad.SERCA2a: 231+/-54, n=24). Using AP voltage-clamping, we recorded a 0.2 mM Cd-sensitive current which can be ascribed to Ca current flowing during the AP. The integral of this current was reduced in Ad.SERCA2a myocytes compared with control (non-infected charge (pC): 27.5+/-4.2, n=8; Ad.SERCA2a: 15.5+/-4.1, n=11; P<0.01). Using AP clamping during the loading protocol, to take into account changes in APD, SR Ca content (assessed by integrating a 20 mM caffeine-induced inward current) was significantly larger in Ad.SERCA2a compared with both controls (SR Ca content (microM/l non-mitochondrial volume): non-infected: 25.5+/-7, n=8; Ad.GFP: 25.7+/-11, n=6; Ad.SERCA2a: 80.5+/-19, n=8). In conclusion, this study shows that SR Ca content is increased despite decreased Ca entry after overexpression of SERCA2a, and this can lead to positive inotropism. This effect coupled with shorter APD may be a useful therapeutic modality in heart failure. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12098219&query_hl=1 ER - TY - JFULL T1 - Specific beta(2)AR blocker ICI 118,551 actively decreases contraction through a G(i)-coupled form of the beta(2)AR in myocytes from failing human heart. A1 - Gong, H A1 - Sun, H A1 - Koch, WJ A1 - Rau, T A1 - Eschenhagen, T A1 - Ravens, U A1 - Heubach, JF A1 - Adamson, DL A1 - Harding, SE J1 - Circulation Y1 - 2002/05/28/ VL - 105 SN - 1524-4539 SP - 2497 EP - 2503 N2 - BACKGROUND: We have observed direct (noncatecholamine-blocking) negative inotropic effects of the selective beta(2)-adrenoceptor (AR) antagonist ICI 118,551 in myocytes from failing human ventricle. In this study we characterize the effect in parallel in human myocytes and in myocytes from animal models where beta(2)ARs or G(i) proteins are overexpressed. METHODS AND RESULTS: Enzymatically isolated, superfused ventricular myocytes were exposed to betaAR agonists and antagonists/inverse agonists, and contraction amplitude was measured. ICI 118,551 decreased contraction in ventricular myocytes from failing human hearts by 45.3+/-4.1% (n=20 hearts/31 myocytes, P<0.001) but had little effect in nonfailing hearts (4.9+/-4%, n=5 myocytes/3 hearts). Effects were significantly larger in patients classified as end-stage. Transgenic mice with high beta(2)AR number and increased G(i) levels had normal basal contractility but showed a similar negative inotropic response to ICI 118,551. Overexpression of human beta(2)AR in rabbit myocytes using adenovirus potentiated the negative inotropic effect of ICI 118,551. In human, rabbit, and mouse myocytes, the negative inotropic effects were blocked after treatment of cells with pertussis toxin to inactivate G(i), and overexpression of G(i)alpha(2) induced the effect de novo in normal rat myocytes. CONCLUSIONS: We hypothesize that ICI 118,551 binding directs the beta(2)AR to a G(i)-coupled form and away from the G(s)-coupled form (ligand-directed trafficking). ICI 118,551 effectively acts as an agonist at the G(i)-coupled beta(2)AR, producing a direct negative inotropic effect. Conditions where beta(2)ARs are present and G(i) is raised (failing human heart, TGbeta(2) mouse heart) predispose to the appearance of the negative inotropic effect. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12034656&query_hl=1 ER - TY - JFULL T1 - Association of tapasin and COPI provides a mechanism for the retrograde transport of major histocompatibility complex (MHC) class I molecules from the Golgi complex to the endoplasmic reticulum. A1 - Paulsson, KM A1 - Kleijmeer, MJ A1 - Griffith, J A1 - Jevon, M A1 - Chen, S A1 - Anderson, PO A1 - Sjogren, HO A1 - Li, S A1 - Wang, P J1 - J Biol Chem Y1 - 2002/05/24/ VL - 277 SN - 0021-9258 SP - 18266 EP - 18271 N2 - Tapasin is a subunit of the transporter associated with antigen processing (TAP). It associates with the major histocompatibility complex (MHC) class I. We show that tapasin interacts with beta- and gamma-subunits of COPI coatomer. COPI retrieves membrane proteins from the Golgi network back to the endoplasmic reticulum (ER). The COPI subunit-associated tapasin also interacts with MHC class I molecules suggesting that tapasin acts as the cargo receptor for packing MHC class I molecules as cargo proteins into COPI-coated vesicles. In tapasin mutant cells, neither TAP nor MHC class I are detected in association with the COPI coatomer. Interestingly, tapasin-associated MHC class I molecules are antigenic peptide-receptive and detected in both the ER and the Golgi. Our data suggest that tapasin is required for the COPI vesicle-mediated retrograde transport of immature MHC class I molecules from the Golgi network to the ER. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11884415&query_hl=1 ER - TY - JFULL T1 - Signal transduction pathways for the expression of genes for MMP and TIMP induced by stretching human fetal scleral fibroblasts A1 - Cui, W A1 - Bryant, MR A1 - McDonnell, PJ J1 - INVEST OPHTH VIS SCI Y1 - 2002/05// VL - 43 SN - 0146-0404 SP - U582 EP - U582 ER - TY - JFULL T1 - High-resolution scanning patch-clamp: new insights into cell function. A1 - Gu, Y A1 - Gorelik, J A1 - Spohr, HA A1 - Shevchuk, A A1 - Lab, MJ A1 - Harding, SE A1 - Vodyanoy, I A1 - Klenerman, D A1 - Korchev, YE J1 - FASEB J Y1 - 2002/05// VL - 16 SN - 1530-6860 SP - 748 EP - 750 N2 - Cell specialization is often governed by the spatial distribution of ion channels and receptors on the cell surface. So far, little is known about functional ion channel localization. This is due to a lack of satisfactory methods for investigating ion channels in an intact cell and simultaneously determining the channels' positions accurately. We have developed a novel high-resolution scanning patch-clamp technique that enables the study of ion channels, not only in small cells, such as sperm, but in submicrometer cellular structures, such as epithelial microvilli, fine neuronal dendrites, and, particularly, T-tubule openings of cardiac myocytes. In cardiac myocytes, as in most excitable cells, action potential propagation depends essentially on the properties of ion channels that are functionally and spatially coupled. We found that the L-type calcium and chloride channels are distributed and colocalized in the region of T-tubule openings, but not in other regions of the myocyte. In addition, chloride channels were found in narrowly defined regions of Z-grooves. This finding suggests a new synergism between these types of channels that may be relevant for action potential propagation along the T-tubule system and excitation-contraction coupling. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11923226&query_hl=1 ER - TY - JFULL T1 - Estimating the noise mitigation effect of local decoupling in printed circuit boards A1 - Fan, J A1 - Cui, W A1 - Drewniak, JL A1 - Van Doren, TP A1 - Knighten, JL J1 - IEEE T ADV PACKAGING Y1 - 2002/05// VL - 25 SN - 1521-3323 SP - 154 EP - 165 N2 - Local decoupling, i.e., placing decoupling capacitors sufficiently close to device power/ground pins in order to decrease the impedance of power bus at frequencies higher than the series resonant frequency, has been studied using a modeling approach, a hybrid lumped/distributed circuit model established and an expression to quantify the benefits of power bus noise mitigation due to local decoupling developed. In this work, a test board with a local decoupling capacitor was studied and the noise mitigation effect due to the capacitor placed adjacent to an imput test port was measured. Closed-form expressions for self and mutual inductances of vias are developed, so that the noise mitigation effect can then be estimated using the previously developed expression. The difference between the estimates and measurements is approximately 1 dB, which demonstrates the application of these closed-form expressions in the PCB power bus designs. Shared-via decoupling, capacitors sharing vias with device power/ground pins, is also modeled as an extreme case of local decoupling. ER - TY - JFULL T1 - MafB is expressed in developing lens and cooperates with Pax6 and Prox1 to regulate chicken beta B1 crystallin gene expression A1 - Cui, W A1 - Tomarev, SI A1 - Chepelinsky, AB A1 - Duncan, MK J1 - INVEST OPHTH VIS SCI Y1 - 2002/05// VL - 43 SN - 0146-0404 SP - U555 EP - U555 ER - TY - JFULL T1 - Leucocyte-endothelium interaction in the symptomatic carotid plaque A1 - Clayton, GS A1 - Adams, J A1 - Golledge, J A1 - Nohadani, MR A1 - Bishop, AE A1 - Greenhalgh, RM A1 - Davies, AH J1 - BRIT J SURG Y1 - 2002/05// VL - 89 SN - 0007-1323 SP - 636 EP - 636 ER - TY - JFULL T1 - A nerve-sparing radical hysterectomy: guidelines and feasibility in Western patients. A1 - Barton, DP A1 - Butler-Manuel, SA A1 - Buttery, LD A1 - A'Hern, RP A1 - Polak, JM J1 - Int J Gynecol Cancer Y1 - 2002/05// VL - 12 SN - 1048-891X SP - 319; author reply 321 EP - 319; author reply 321 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12060457&query_hl=1 ER - TY - JFULL T1 - Cloning and tissue distribution of three murine alpha/beta hydrolase fold protein cDNAs. A1 - Edgar, AJ A1 - Polak, JM J1 - Biochem Biophys Res Commun Y1 - 2002/04/05/ VL - 292 SN - 0006-291X SP - 617 EP - 625 N2 - We have cloned 3 novel murine cDNAs encoding proteins containing an alpha/beta hydrolase fold; a catalytic domain found in a very wide range of enzymes. These proteins belong to the prosite UPF0017 uncharacterized protein family and we have named them lung alpha/beta hydrolase 1, 2, and 3 (LABH) since they were cloned from lung cDNA. All have 9 coding exons, encoding 412, 425, and 411 residue proteins respectively (46-48 kDa); LABH1 being closely related to LABH3 having 45% identity. All 3 proteins have a single predicted amino-terminus transmembrane domain. An alignment of family members from different phyla enabled the identification of the LABH1 catalytic triad as Ser211, Asp337, and His366. mRNA expression levels of LABH1 and 3 were highest in liver and LABH2 highest in testis. These findings suggest that the LABH proteins consist of a novel family of membrane bound enzymes whose function has yet to be determined. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11922611&query_hl=1 ER - TY - JFULL T1 - Evidence of an environmental cause for the rise in cholangiocarcinoma A1 - Khan, SA A1 - Carmichael, PL A1 - Taylor-Robinson, SD A1 - Habib, N A1 - Thomas, HC J1 - GUT Y1 - 2002/04// VL - 50 SN - 0017-5749 SP - A54 EP - A54 ER - TY - JFULL T1 - Reciprocal age-related changes in GAP-43/B-50, substance P and calcitonin gene-related peptide (CGRP) expression in rat primary sensory neurones and their terminals in the dorsal horn of the spinal cord and subintima of the knee synovium. A1 - Hukkanen, M A1 - Platts, LA A1 - Corbett, SA A1 - Santavirta, S A1 - Polak, JM A1 - Konttinen, YT J1 - Neurosci Res Y1 - 2002/04// VL - 42 SN - 0168-0102 SP - 251 EP - 260 N2 - Age-related changes in the expression of the growth associated protein GAP-43/B-50, and the neuropeptides substance P and calcitonin gene-related peptide (CGRP) were investigated in the sensory neurones of rat dorsal root ganglia, dorsal horns of the spinal cord and subintimal knee synovium. The two time-points studied were 2 months (young adults) and 14-month (aged)-old Sprague Dawley rats. Dorsal root ganglia: In young adults, 40 and 35% of the L4-L5 dorsal root ganglion neurones were positive for GAP-43/B-50 with a 1.5 fold increase in frequency in aged rats at the L5 ganglion. GAP-43/B-50 was strongly expressed by the non-neuronal satellite cells of some medium and many large sized neurones in aged rats. There were marked reciprocal shifts between small and medium sized sensory neurones in respect to their substance P and CGRP expression profiles. Dorsal horn of the spinal cord: there was a 1.3 fold decrease of substance P at L5 level and a 1.3 and 1.5 fold decrease of CGRP at L4-L5 levels in aged rats, respectively. Synovial membrane: There was a 2.3 fold increase in GAP-43/B-50 and a 2.5 fold decrease of CGRP with no changes in substance P expression. These results indicate that (i) primary sensory neurones undergo age-related changes already in early stages of aging, (ii) aging may result in a reduction of substance P and CGRP axonal transport, and (iii) reduced numbers of CGRP containing synovial perivascular fibres may imply a deficient regulation of the synovial microvasculature and therefore metabolic homeostasis of the joint in aged subjects. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11985877&query_hl=1 ER - TY - JFULL T1 - Effects of Na(+)/Ca(2+)-exchanger overexpression on excitation-contraction coupling in adult rabbit ventricular myocytes. A1 - Ranu, HK A1 - Terracciano, CM A1 - Davia, K A1 - Bernobich, E A1 - Chaudhri, B A1 - Robinson, SE A1 - Bin Kang, Z A1 - Hajjar, RJ A1 - MacLeod, KT A1 - Harding, SE J1 - J Mol Cell Cardiol Y1 - 2002/04// VL - 34 SN - 0022-2828 SP - 389 EP - 400 N2 - The Na(+)/Ca(2+)-exchanger (NCX) is the main mechanism by which Ca(2+) is transported out of the ventricular myocyte. NCX levels are raised in failing human heart, and the consequences of this for excitation-contraction coupling are still debated. We have increased NCX levels in adult rabbit myocytes by adenovirally-mediated gene transfer and examined the effects on excitation-contraction coupling after 24 and 48 h. Infected myocytes were identified through expression of green fluorescent protein (GFP), transfected under a separate promoter on the same viral construct. Control experiments were done with both non-infected myocytes and those infected with adenovirus expressing GFP only. Contraction amplitude was markedly reduced in NCX-overexpressing myocytes at either time point, and neither increasing frequency nor raising extracellular Ca(2+) could reverse this depression. Resting membrane potential and action potential duration were largely unaffected by NCX overexpression, as was peak Ca(2+) entry via the L-type Ca(2+) channel. Systolic and diastolic Ca(2+) levels were significantly reduced, with peak systolic Ca(2+) in NCX-overexpressing myocytes lower than diastolic levels in control cells at 2 m m extracellular Ca(2+). Both cell relengthening and the decay of the Ca(2+) transient were significantly slowed. Sarcoplasmic reticulum (SR) Ca(2+) stores were completely depleted in a majority of myocytes, and remained so despite increasingly vigorous loading protocols. Depressed contractility following NCX overexpression is therefore related to decreased SR Ca(2+) stores and low diastolic Ca(2+) levels rather than reduced Ca(2+) entry. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11991729&query_hl=1 ER - TY - JFULL T1 - Classical and novel steroid actions: a unified but complex view. A1 - Valverde, MA A1 - Parker, MG J1 - Trends Biochem Sci Y1 - 2002/04// VL - 27 SN - 0968-0004 SP - 172 EP - 173 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11943538&query_hl=1 ER - TY - JFULL T1 - Tissue engineering and ENT surgery. A1 - Patel, NN A1 - Butler, PE A1 - Buttery, L A1 - Polak, JM A1 - Tolley, NS J1 - J Laryngol Otol Y1 - 2002/03// VL - 116 SN - 0022-2151 SP - 165 EP - 169 N2 - Tissue engineering is the development of biological substitutes for the repair and regeneration of damaged tissues. We explain the principles of this emerging field of biotechology. The present and potential applications of tissue engineering technologies in ENT surgery are then reviewed. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11893255&query_hl=1 ER - TY - JFULL T1 - Recent insights into the pathogenesis and therapeutics of pulmonary hypertension. A1 - Strange, JW A1 - Wharton, J A1 - Phillips, PG A1 - Wilkins, MR J1 - Clin Sci (Lond) Y1 - 2002/03// VL - 102 SN - 0143-5221 SP - 253 EP - 268 N2 - The normal adult pulmonary circulation is a low-pressure, high-capacity circuit. Pulmonary vascular resistance is regulated by alveolar oxygen tension, potassium channels and a variety of locally produced and circulating vasoactive factors. Perturbations of these systems may contribute to the pathogenesis of pulmonary hypertension. Recently, mutations in BMPR2 and ALK-1, genes that encode members of the transforming growth factor-beta (TGF-beta) receptor superfamily, have been found in patients with primary pulmonary hypertension. These observations provide a novel insight into the pathogenesis of primary pulmonary hypertension, and emphasize the importance of the integrity of the TGF-beta receptor family in the maintenance of normal pulmonary vascular structure and function. This review discusses the latest developments in the field of pulmonary vascular biology and the prospects for improving the treatment of pulmonary hypertension. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11869166&query_hl=1 ER - TY - JFULL T1 - Gender- and age-related differences in osteoclast formation from circulating precursors. A1 - Jevon, M A1 - Sabokbar, A A1 - Fujikawa, Y A1 - Hirayama, T A1 - Neale, SD A1 - Wass, J A1 - Athanasou, NA J1 - J Endocrinol Y1 - 2002/03// VL - 172 SN - 0022-0795 SP - 673 EP - 681 N2 - A number of bone diseases characterised by excessive osteolysis (e.g. osteoporosis and Paget's disease) exhibit a marked gender difference in prevalence and are more common in the elderly population. Bone resorption is carried out by osteoclasts, which are formed by fusion of circulating mononuclear precursor cells of haematopoietic origin. In this study, we have determined whether there are gender- and age-related differences in osteoclast formation from circulating precursors. Peripheral blood mononuclear cells (PBMCs) were co-cultured with UMR106 osteoblast-like cells in the presence of macrophage-colony stimulating factor (M-CSF) and 1,25 dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) or cultured alone in the presence of sRANKL (soluble receptor activator of nuclear factor kappa B ligand) and M-CSF. As assessed by the formation of tartrate resistant acid phosphatase (TRAP)-positive (TRAP(+)) and vitronectin receptor-positive (VNR(+)) multinucleated cells (MNCs), there was no difference in the number of circulating osteoclast precursors in males and females. Lacunar resorption carried out by osteoclasts formed from these precursors was generally increased in males compared with females (P=0.03). An increase in the number of TRAP(+) and VNR(+) MNCs formed from male PBMCs was noted in response to 1,25(OH)(2)D(3) (P<0.005). An increase in lacunar resorption in cultures of PBMCs (10(5) per well) from males was also noted in response to 10(-9) M 1,25(OH)(2)D(3) (P<0.05) and sRANKL (P=0.05), but not M-CSF. The addition of dexamethasone resulted in a marked increase in osteoclast formation and lacunar resorption in both males and females. Post-menopausal females and males of comparable age showed similar levels of osteoclastogenesis. Pre-menopausal women showed similar levels of osteoclastogenesis but less resorption (P=0.01) compared with males of comparable age. These results show that there are specific gender/age-related differences in osteoclast formation and bone resorption and have implications for evaluating osteoclastogenesis in skeletal diseases such as primary osteoporosis and Paget's disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11874715&query_hl=1 ER - TY - JFULL T1 - Targeting phospholamban by gene transfer in human heart failure. A1 - del Monte, F A1 - Harding, SE A1 - Dec, GW A1 - Gwathmey, JK A1 - Hajjar, RJ J1 - Circulation Y1 - 2002/02/26/ VL - 105 SN - 1524-4539 SP - 904 EP - 907 N2 - BACKGROUND: Myocardial cells from failing human hearts are characterized by abnormal calcium handling, a negative force-frequency relationship, and decreased sarcoplasmic reticulum Ca2+ ATPase (SERCA2a) activity. In this study, we tested whether contractile function can be improved by decreasing the inhibitory effects of phospholamban on SERCA2a with adenoviral gene transfer of antisense phospholamban (asPL). METHODS AND RESULTS: Myocardial cells isolated from 9 patients with end-stage heart failure and 18 donor nonfailing hearts were infected with adenoviruses encoding for either the antisense of phospholamban (Ad.asPL), the SERCA2a gene (Ad.SERCA2a), or the reporter genes beta-galactosidase and green fluorescent protein (Ad.betagal-GFP). Adenoviral gene transfer with Ad.asPL decreased phospholamban expression over 48 hours, increasing the velocity of both contraction and relaxation. Compared with cardiomyocytes infected with Ad.asPL (n=13), human myocytes infected with Ad.betagal-GFP (n=8) had enhanced contraction velocity (20.3 +/- 3.9% versus 8.7 +/- 2.6% shortening/second; P<0.01) and relaxation velocity (26.0 +/- 6.2% versus 8.6 +/- 4.3% shortening/second; P<0.01). The improvement in contraction and relaxation velocities was comparable to cardiomyocytes infected with Ad.SERCA2a. Failing human cardiomyocytes had decreased contraction and Ca2+ release with increasing frequency (0.1 to 2 Hz). Phospholamban ablation restored the frequency response in the failing cardiomyocytes to normal; increasing frequency resulted in enhanced sarcoplasmic reticulum Ca2+ release and contraction. CONCLUSION: These results show that gene transfer of asPL can improve the contractile function in failing human myocardium. Targeting phospholamban may provide therapeutic benefits in human heart failure. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11864915&query_hl=1 ER - TY - JFULL T1 - Eotaxin-2 generation is differentially regulated by lipopolysaccharide and IL-4 in monocytes and macrophages. A1 - Watanabe, K A1 - Jose, PJ A1 - Rankin, SM J1 - J Immunol Y1 - 2002/02/15/ VL - 168 SN - 0022-1767 SP - 1911 EP - 1918 N2 - The eotaxins are a family of CC chemokines that coordinate the recruitment of inflammatory cells, in particular eosinophils, to sites of allergic inflammation. The cDNA for eotaxin-2 (CC chemokine ligand 24) was originally isolated from an activated monocyte library. In this study, we show for the first time that peripheral blood monocytes generate bioactive eotaxin-2 protein constitutively. Eotaxin-2 production was significantly up-regulated when monocytes were stimulated with the proinflammatory cytokine IL-1beta and the microbial stimuli, LPS and zymosan. In contrast, the Th2 cytokines, IL-4 and IL-13, and the proinflammatory cytokine, TNF-alpha, acting alone or in combination, did not enhance the generation of eotaxin-2 by monocytes. Indeed, IL-4 suppressed the generation of eotaxin-2 by LPS-stimulated monocytes. Although other chemokines, including macrophage-inflammatory protein-1alpha, monocyte chemoattractant protein-1, macrophage-derived chemokine, and IL-8 were generated by monocytes, eotaxin-1 (CC chemokine ligand 11) could not be detected in the supernatants of monocytes cultured in the presence or absence of any of the stimuli used in the above experiments. Furthermore, human dermal fibroblasts that produce eotaxin-1 did not generate eotaxin-2 under basal conditions or when stimulated with specific factors, including IL-4, IL-13, TNF-alpha, and LPS. When monocytes were differentiated into macrophages, their constitutive generation of eotaxin-2 was suppressed. Moreover, IL-4, but not LPS, up-regulated the production of eotaxin-2 by macrophages. Taken as a whole, these results support a role for macrophage-derived eotaxin-2 in adaptive immunity, with a Th2 bias. In contrast, a role for monocyte-derived eotaxin-2 is implicated in innate immunity. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11823526&query_hl=1 ER - TY - JFULL T1 - Third-generation biomedical materials. A1 - Hench, LL A1 - Polak, JM J1 - Science Y1 - 2002/02/08/ VL - 295 SN - 1095-9203 SP - 1014 EP - 1017 N2 - Whereas second-generation biomaterials were designed to be either resorbable or bioactive, the next generation of biomaterials is combining these two properties, with the aim of developing materials that, once implanted, will help the body heal itself. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11834817&query_hl=1 ER - TY - JFULL T1 - A comparison of cell and tissue extraction techniques using high-resolution 1H-NMR spectroscopy. A1 - Le Belle, JE A1 - Harris, NG A1 - Williams, SR A1 - Bhakoo, KK J1 - NMR Biomed Y1 - 2002/02// VL - 15 SN - 0952-3480 SP - 37 EP - 44 N2 - Analysis of brain metabolites by a wide range of analytical techniques is typically achieved using biochemical extraction methodologies that require either two separate samples or two separate extraction steps to prepare both aqueous and organic metabolite fractions. However there are a number of brain pathologies in which both aqueous metabolite and lipid changes occur so that a simultaneous extraction of both fractions would be valuable. The methanol-chloroform (M/C) technique enables extraction of both aqueous metabolites and lipids simultaneously. It is already well established for lipid extraction of cells and tissue but its efficiency and reproducibility for extraction of aqueous metabolites is unknown. Therefore, we compared the aqueous metabolite yield and the reproducibility of the M/C method to the commonly used perchloric acid (PCA) method, using 1H-NMR spectroscopy of adult rat brain and purified rat astrocyte culture extracts. The results indicate that M/C is a superior technique for aqueous metabolite extraction from both brain tissue and cells when compared to the PCA method. The M/C extraction technique enables the simultaneous extraction of both lipids and aqueous metabolites from a single sample using small solvent-volumes, making it well suited for NMR investigations of both tissues and cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11840551&query_hl=1 ER - TY - JFULL T1 - Steps toward mapping the human vasculature by phage display A1 - Arap, W A1 - Kolonin, MG A1 - Trepel M A1 - Lahdenranta J A1 - Cardo-Vila M A1 - Giordano RJ A1 - Mintz PJ A1 - Ardelt, PU A1 - Yao, VJ A1 - Vidal, CI A1 - Chen, L A1 - Flamm, A A1 - Valtanen, H A1 - Weavind, LM A1 - Hicks, ME A1 - Pollock, RE A1 - Botz, GH A1 - Bucana, CD A1 - Koivunen, E A1 - Cahill, D A1 - Troncoso, P A1 - Baggerly, KA A1 - Pentz, RD A1 - Do, KA A1 - Logothetis, CJ A1 - Pasqualini, R J1 - Nat Med. Y1 - 2002/02// IS - 2 VL - 8 SN - 1078-8956 SP - 121 EP - 127 ER - TY - JFULL T1 - Embryo transfer experiments and ovarian transplantation identify the ovary as the only site in which nuclear receptor interacting protein 1/RIP140 action is crucial for female fertility. A1 - Leonardsson, G A1 - Jacobs, MA A1 - White, R A1 - Jeffery, R A1 - Poulsom, R A1 - Milligan, S A1 - Parker, M J1 - Endocrinology Y1 - 2002/02// VL - 143 SN - 0013-7227 SP - 700 EP - 707 N2 - Spatial and temporal regulation of gene expression by a number of different nuclear receptors is critical in female reproduction. In this study we investigated whether the nuclear receptor corepressor nuclear receptor interacting protein 1 (Nrip1)/RIP140, which is essential for ovulation, is also required for postovulatory events, leading to pregnancy and parturition. Expression analysis indicated that Nrip1 is present in the uterus in stromal and glandular epithelial cells, primary decidual cells, and subsequently in differentiating decidual cells at the anti-mesometrial side of the implantation site. It also indicated a temporal regulation of Nrip1 in the corpora lutea at different stages of pregnancy, with increased levels at midgestation at approximately d 9.5 postcoitum (pc). By performing both embryo and ovarian transfer experiments we demonstrate that, provided the block to ovulation is by-passed, Nrip1(-/-) mice are capable of establishing and maintaining pregnancies. However, although the majority of offspring derived from ovarian transplantation survived, approximately 50% of embryos were resorbed by d 13.5 pc after embryo transfer, and the majority of pups were stillborn or died soon thereafter. Thus, although Nrip1 is differentially expressed in the reproductive tract, we conclude that the ovary is the only site in which its action is essential for fertility, with a crucial role in ovulation and a secondary role in the maintenance of pregnancy. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11796527&query_hl=1 ER - TY - JFULL T1 - Altered mechanical properties and intracellular calcium signaling in cardiomyocytes from annexin 6 null-mutant mice A1 - Song, GJ A1 - Harding, SE A1 - Duchen, MR A1 - Tunwell, R A1 - O'Gara, P A1 - Hawkins, TE A1 - Moss, SE J1 - FASEB J Y1 - 2002/02// VL - 16 SN - 0892-6638 SP - 622 EP - + N2 - Annexin 6 is one of a widely expressed family of calcium-binding proteins found in most mammalian tissues, including the heart. Several studies have implicated annexin 6 in the regulation of intracellular Ca2+ signaling, and it has been shown in vitro to act as a modulator of the sarcoplasmic reticulum Ca2+-release channel, cardiac L-type calcium channel, and Na+/Ca2+ exchanger. To investigate the role of annexin 6 in intact cardiomyocytes, we used mice containing a targeted disruption of the annexin 6 gene. Compared with controls, the myocytes of annexin 6 null-mutant mice demonstrated a significant increase in the rates of shortening and relengthening. Intracellular Ca2+ transients in fura-2-loaded cardiomyocytes induced by caffeine showed a normal baseline and amplitude, whereas the rate of decay was doubled in annexin 6(-/-) myocytes compared with control mice. These results show that annexin 6 knockout in the mouse leads to an increase in myocyte contractility and faster diastolic Ca2+ removal from the cytoplasm. In light of published findings showing annexin 6 to be down-regulated in end-stage heart failure, these results are consistent with a role for annexin 6 as a negative inotropic factor in the regulation of cardiomyocyte mechanics. ER - TY - JFULL T1 - ET(A) and ET(B) receptors modulate the proliferation of human pulmonary artery smooth muscle cells. A1 - Davie, N A1 - Haleen, SJ A1 - Upton, PD A1 - Polak, JM A1 - Yacoub, MH A1 - Morrell, NW A1 - Wharton, J J1 - Am J Respir Crit Care Med Y1 - 2002/02/01/ VL - 165 SN - 1073-449X SP - 398 EP - 405 N2 - We determined the distribution of ET(A) and ET(B) receptors in pulmonary arteries from pulmonary hypertensive patients and control subjects, using in vitro autoradiography, and investigated their role in mediating the proliferative effects of endothelin-1 (ET-1) on distal human pulmonary artery smooth muscle cells (PASMCs). Distal arteries possessed more medial [(125)I]-ET-1 binding sites (105 +/- 10 versus 45 +/- 6 amol/mm(2); p < 0.001) and a greater proportion of ET(B) receptors than proximal arteries (36 +/- 3% versus 3 +/- 1%; p < 0.001). Receptor density in distal arteries and lung parenchyma was twofold greater (p < 0.05) in pulmonary hypertensive patients than in control subjects. ET-1 (10(-9)-10(-7) mol/L) stimulated DNA synthesis (147 +/- 10% of control subjects; p < 0.05) and attenuated the antiproliferative action of cicaprost and forskolin on PASMCs, these effects being mediated via ET(A) and ET(B) receptors. Serum-stimulated proliferation was attenuated by inhibiting either endogenous ET-1 release with phosphoramidon (10(-5) mol/L) or its action with PD145065 (10(-5) mol/L). Cicaprost (10(-10)-10(-7) mol/L) inhibited ET-1 release from PASMCs (49 +/- 16% of control after 24 h; p < 0.001) and increased intracellular cAMP levels, whereas ET(B) receptor stimulation selectively reduced cAMP levels. In conclusion, ET(A) and ET(B) receptors are differentially distributed in human pulmonary arteries. Both receptors promote the proliferation of PASMCs in vitro and may contribute to vascular remodeling in pulmonary hypertension. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11818328&query_hl=1 ER - TY - JFULL T1 - Swelling-activated taurine and creatine effluxes from rat cortical astrocytes are pharmacologically distinct. A1 - Bothwell, JH A1 - Styles, P A1 - Bhakoo, KK J1 - J Membr Biol Y1 - 2002/01/15/ VL - 185 SN - 0022-2631 SP - 157 EP - 164 N2 - Primary cultures of rat cortical astrocytes undergo a swelling-activated loss of taurine and creatine. In this study, the pharmacological characteristics of the taurine and creatine efflux pathways were compared, and significant differences were shown to exist between the two. Both taurine and creatine effluxes were rapidly activated upon exposure of astrocytes to hypo-osmotic media, and rapidly inactivated upon their return to iso-osmotic media. The relative rates of taurine and creatine efflux depended upon the magnitude of the hypo-osmotic shock. Anion-transport inhibitors strongly inhibited taurine efflux, with the order of potency being NPPB > DIDS > niflumic acid. DIDS and NPPB had less of an inhibitory effect on creatine efflux, whereas tamoxifen and niflumic acid actually stimulated creatine efflux. These data are consistent with separate pathways for taurine and creatine loss during astrocyte swelling. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11891574&query_hl=1 ER - TY - JFULL T1 - Effects of combinations of therapeutic agents on the proliferation of progenitor cells in chronic myeloid leukaemia. A1 - Marley, SB A1 - Davidson, RJ A1 - Goldman, JM A1 - Gordon, MY J1 - Br J Haematol Y1 - 2002/01// VL - 116 SN - 0007-1048 SP - 162 EP - 165 N2 - Combination of STI571, a tyrosine kinase inhibitor, with other drugs may be beneficial in the treatment of chronic myeloid leukaemia (CML). We measured the effects of STI571, AG490, farnesyltransferase inhibitor (FTI), interferon alpha (IFN-alpha), cytosine arabinoside (Ara-C) and all-trans retinoic acid (ATRA), singly and in combination, on clonogenic leukaemic cell proliferation. STI571, IFN-alpha and ATRA each reduced proliferation by 50-60%; AG490, FTI and Ara-C had less effect. Comparing the observed and expected (i.e. additive) effects of drug combinations showed STI571 + FTI, STI571 + AG490 and IFN-alpha + ATRA were additive; STI571 + IFN-alpha, IFN-alpha + Ara-C and STI571 + AG490 + FTI were less than additive. Thus, STI571 + FTI, STI571 + AG490 and IFN-alpha + ATRA may be better combination therapies for CML than STI571 + IFN-alpha, IFN-alpha + Ara-C or STI571 + AG490 + FTI. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11841411&query_hl=1 ER - TY - JFULL T1 - Inhibition of myc-dependent breast tumor formation in transgenic mice. A1 - Cui, W A1 - Gusterson, BA A1 - Clark, AJ J1 - Breast Cancer Res Treat Y1 - 2002/01// VL - 71 SN - 0167-6806 SP - 9 EP - 20 N2 - One of the most promising approaches for cancer gene therapy is the use of the so-called suicide genes, which encode prodrug-activating enzymes and render transduced cells more sensitive to prodrugs. The enzyme nitroreductase (NTR) converts prodrug CB1954 into a cytotoxic DNA interstrand cross-linking agent. We have established transgenic mice in which the pro-oncogene c-myc and NTR were fused to the internal ribosome entry site and coexpressed in luminal cells of the mammary gland under the control of mouse whey acidic protein (WAP) promoter to evaluate NTR mediated ablation of mammary tumors. More than 78% of transgenic females developed in situ or infiltrating carcinomas after three to four pregnancies. By contrast, if the transgenic female mice were given the prodrug CB1954 during their third lactation, the incidence of tumors decreased to less than 40% (P < 0.05). The total number of carcinomas was even more striking with 117 carcinomas identified in 14 non-ablated transgenics compared with only five in 15 treated animals (p < 0.05, student t test). C-myc induced pleomorphic nuclei and mitotic figures were seen as a field change in over 70% of the untreated transgenics compared to 20% in the treated group. Our results suggest that the enzyme pro-drug system NTR-CB1954 efficiently inhibit myc-dependent tumor formation and malignant progression in the mammary gland. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11859878&query_hl=1 ER - TY - JFULL T1 - In situ characterisation of living cells by Raman spectroscopy A1 - Notingher, I A1 - Verrier, S A1 - Romanska, H A1 - Bishop, AE A1 - Polak, JM A1 - Hench, LL J1 - SPECTROSC-INT J Y1 - 2002/// VL - 16 SN - 0712-4813 SP - 43 EP - 51 N2 - We report the first Raman spectra of individual living and dead cells (MLE-12 line) cultured on bioinert standard poly-L-lysine coated fused silica and on bioactive 45S5 Bioglass((R)) measured at 785 nm laser excitation. At this excitation wavelength no damage was induced to the cells even after 40 minutes irradiation at 115 mW power, as indicated by cell morphology observation and trypan blue viability test. We show that shorter wavelength lasers, 488 nm and 514 nm, cannot be used because they induce damage to the cells at very low laser powers (5 mW) and short irradiation times (5-20 minutes). The most important differences between the spectra of living and dead cells are in the 1530-1700 cm(-1) range, where the dead cells have strong peaks at 1578 cm(-1) and 1607 cm(-1). Other differences occur around the DNA peak at 1094 cm(-1). Our study establishes the feasibility of using the 785 nm laser for an in situ real-time non-invasive method to follow biological events (proliferation, differentiation, cell death, etc.) within individual cells cultured on bioactive scaffolds in their physiologic environment over long periods of time. ER - TY - JFULL T1 - In situ characterisation of living cells by Raman spectroscopy A1 - Nothinger I A1 - Verrier S A1 - Romanska HM A1 - Bishop AE A1 - Polak JM A1 - Hench LL J1 - Spectroscopy Y1 - 2002/// VL - 16 SN - 0712-4813 SP - 43 EP - 51 ER - TY - JFULL T1 - Repetitive myocardial stunning in pigs is associated with an increased formation of reactive nitrogen species. A1 - Baker, CS A1 - Frost, MT A1 - Rimoldi, O A1 - Moore, K A1 - Halliwell, B A1 - Polak, JM A1 - Camici, PG A1 - Hall, RJ J1 - Heart Y1 - 2002/01// VL - 87 SN - 1468-201X SP - 77 EP - 78 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11751673&query_hl=1 ER - TY - JFULL T1 - Pelvic nerve plexus trauma at radical and simple hysterectomy: a quantitative study of nerve types in the uterine supporting ligaments. A1 - Butler-Manuel, SA A1 - Buttery, LD A1 - A'Hern, RP A1 - Polak, JM A1 - Barton, DP J1 - J Soc Gynecol Investig Y1 - 2002/01// VL - 9 SN - 1071-5576 SP - 47 EP - 56 N2 - OBJECTIVE: Using neuropeptide and enzyme markers to autonomic nerves, we sought to demonstrate and quantify the nerve types contained within the uterosacral ligaments (USLs) and cardinal ligaments (CLs) that are divided during radical hysterectomy (RH). METHODS: Cross-sectional biopsies were collected from the lateral third of the USL and the CL in 24 women who had an RH for cervical cancer, and from the uterine insertion of these ligaments in 11 women who had a simple hysterectomy for benign disease. We applied indirect immunofluorescence with FITC-conjugated secondary antibodies, using polyclonal primary antibodies to neuropeptide markers that predominate within somatic and autonomic nerves, to show different populations of the following nerve types within the biopsies: neuropeptide Y (NPY) and tyrosine hydroxylase (TH) for sympathetic nerves; vasoactive intestinal polypeptide (VIP) for parasympathetic nerves; substance P (SP) for nociceptive and sensory-motor nerves; and calcitonin gene-related peptide (CGRP) for sensory and sensory-motor nerves. The percentage area of immunoreactivity (PAI), determined by a computer-assisted image analyzer attached to a fluorescent microscope, was used as an objective quantitative measure of nerve density. Confocal microscopy was used to determine the composition and spatial arrangement of nerve fibers in the ligaments. RESULTS: The PAI was greater for all markers tested in both the USL and CL (P <.001) in RH compared with simple hysterectomy biopsies. For RH specimens, the PAI was greater for the sympathetic, sensory, and sensory-motor nerve markers in the USL compared with the CL (P <.01), but the PAI for VIP was similar (P >.05). Conversely, excluding the large trunks and associated ganglia, the free nerve fiber PAI in the CL was greater than that of the USL for all nerve markers (P <.001). The staining of peripheral autonomic ganglia and associated fibers, for NPY and TH, indicates that some sympathetic nerves are preganglionic with their cell bodies within the pelvic plexus. CONCLUSIONS: Significantly more autonomic nerves are transected in the more lateral division of the uterine supporting ligaments during a radical hysterectomy than during a simple hysterectomy. Sympathetic, parasympathetic, sensory, and sensory-motor nerve types are present within the CL and USL. The proportions of each nerve type differ between the two ligaments, and sympathetic nerves in the USL are the single largest nerve type. The uterine supporting ligaments are a major pathway for autonomic nerves to the pelvic organs. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11839509&query_hl=1 ER - TY - JFULL T1 - Identification of novel oestrogen receptor target genes in human ZR75-1 breast cancer cells by expression profiling. A1 - Soulez, M A1 - Parker, MG J1 - J Mol Endocrinol Y1 - 2001/12// VL - 27 SN - 0952-5041 SP - 259 EP - 274 N2 - Oligonucleotide microarrays were used to analyse gene expression profiles in human ZR75-1 breast cancer cells in the presence of 17beta-oestradiol and oestrogen antagonists. Differential gene expression of a number of genes was confirmed by quantitative RNA analysis. In addition to known oestrogen-responsive genes, an appreciable number of novel targets were identified, including growth factors and components of the cell cycle, adhesion molecules, enzymes, signalling molecules and transcription factors. The most pronounced oestrogen-sensitive gene was that for the cytochrome P450-IIB enzyme, involved in metabolising steroids and xenobiotics, which was increased 100-fold over a 24 h period. It is a direct target gene for oestrogens, because its expression was increased in the presence of cyclohexamide. In contrast, expression of cytochrome P450-IIB was not detected in human MCF7 breast cancer cells. Expressions of the cationic amino acid transporter E16, gap junction protein and insulin-like growth factor binding protein 4 were also markedly increased by oestrogens, but the kinetics of induction varied according to the target gene. With the exception of the cationic amino acid transporter E16 and the insulin-like growth factor binding protein 4, the expression of the majority of the genes was unaffected by antioestrogen treatment. Further analysis of this set of markers will provide alternative approaches to the investigation of the mitogenicity of oestrogens in breast cancer cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11719280&query_hl=1 ER - TY - JFULL T1 - Association of clonal T-cell large granular lymphocyte disease and paroxysmal nocturnal haemoglobinuria (PNH): further evidence for a pathogenetic link between T cells, aplastic anaemia and PNH A1 - Karadimitris, A A1 - Li, K A1 - Notaro, R A1 - Araten, DJ A1 - Nafa, K A1 - Thertulien, R A1 - Ladanyi, M A1 - Stevens, AE A1 - Rosenfeld, CS A1 - Roberts, IAG A1 - Luzzatto, L J1 - BRIT J HAEMATOL Y1 - 2001/12// VL - 115 SN - 0007-1048 SP - 1010 EP - 1014 N2 - There is mounting evidence to suggest that T-cell-mediated suppression of haemopoiesis is a pathogenetic mechanism in three bone marrow failure syndromes: aplastic anaemia (AA), paroxysmal nocturnal haemobloginuria (PNH) and myelodysplasia (MDS). T-cell microclones can be detected by sensitive polymerase chain reaction (PCR)-based methods in all three disorders. Recently, larger clonal populations of T-cell large granular lymphocytes (T-LGLs) have been observed in some patients with AA and MDS. Here, we report the development of a large clonal T-LGL population in a patient with bona fide PNH. In this patient, we defined part of the sequence of the T-cell receptor (TCR) beta-chain gene, and we have shown that the large T-LGL population emerged from a background of multiple smaller T-cell clones. Thus, T-LGL clones in AA, MDS and PNH probably expand as a result of antigenic stimulation. It is postulated that the antigen driving clonal T-cell proliferations in these disorders exists on haemopoietic stem cells. ER - TY - JFULL T1 - Progress in, and future prospects for, the treatment of primary pulmonary hypertension. A1 - Wilkins, MR A1 - Wharton, J J1 - Heart Y1 - 2001/12// VL - 86 SN - 1468-201X SP - 603 EP - 604 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11711446&query_hl=1 ER - TY - JFULL T1 - Differential expression of carnosine, homocarnosine and N-acetyl-L-histidine hydrolytic activities in cultured rat macroglial cells. A1 - Baslow, MH A1 - Suckow, RF A1 - Berg, MJ A1 - Marks, N A1 - Saito, M A1 - Bhakoo, KK J1 - J Mol Neurosci Y1 - 2001/12// VL - 17 SN - 0895-8696 SP - 351 EP - 359 N2 - N-acetyl-L-histidine (NAH) and N-acetyl-L-aspartate (NAA) are representatives of two series of substances that are synthesized by neurons and other cells in the vertebrate central nervous system (CNS). Histidine containing homologs of NAH are beta-alanyl-L-histidine or carnosine (Carn) and gamma-aminobutyrl-L-histidine or homocarnosine (Hcarn). A homolog of NAA is N-acetylaspartylglutamate (NAAG). These substances belong to a unique group of osmolytes in that they are synthesized in cells that may not to be able to hydrolyze them, and are released in a regulated fashion to a second compartment where they can be rapidly hydrolyzed. In this investigation, the catabolic activities for NAH, Carn, and Hcarn in cultured macroglial cells and neurons have been measured, and the second compartment for NAH and Hcarn has been identified only with astrocytes. In addition, oligodendrocytes can only hydrolyze Carn, although Carn can also be hydrolyzed by astrocytes. Thus, astrocytes express hydrolytic activity against all three substrates, but oligodendrocytes can only act on Carn. The cellular separation of these hydrolytic enzyme activities, and the possible nature of the enzymes involved are discussed. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11859931&query_hl=1 ER - TY - JFULL T1 - Contact-mediated inhibition of hematopoietic progenitor cells resulting from homotypic CD34-CD34 cell adhesion downregulates proteins of the pRb pathway. A1 - Lewis, JL A1 - Nguyen, DX A1 - Wen, BP A1 - Marley, SB A1 - Davidson, J A1 - Goldman, JM A1 - Gordon, MY J1 - BLOOD Y1 - 2001/11/16/ VL - 98 SN - 0006-4971 SP - 84A EP - 84A ER - TY - JFULL T1 - Sialomucin-mediated aggregation and gap junctions in the regulation of hematopoietic progenitor cell renewal. A1 - Marley, SB A1 - Davidson, RJ A1 - Lewis, JL A1 - Goldman, JM A1 - Gordon, MY J1 - BLOOD Y1 - 2001/11/16/ VL - 98 SN - 0006-4971 SP - 293A EP - 293A ER - TY - JFULL T1 - Effects of over-expression of SERCA2a on action potential duration, Ca2+ entry and sarcoplasmic reticulum Ca2+ content in rabbit ventricular myocytes A1 - Chaudhri, B A1 - Hajjar, RJ A1 - Harding, SE A1 - Terracciano, CMN J1 - J PHYSIOL-LONDON Y1 - 2001/11// VL - 536 SN - 0022-3751 SP - 74P EP - 74P ER - TY - JFULL T1 - Progenitor cells from patients with advanced phase chronic myeloid leukaemia respond to STI571 in vitro and in vivo. A1 - Marley, SB A1 - Davidson, RJ A1 - Lewis, JL A1 - Nguyen, DX A1 - Eades, A A1 - Parker, S A1 - Goldman, JM A1 - Gordon, MY J1 - Leuk Res Y1 - 2001/11// VL - 25 SN - 0145-2126 SP - 997 EP - 1002 N2 - STI571 targets p210(BCR-ABL) in chronic myeloid leukaemia (CML). In vitro, STI571 reduces self-replication (replating ability) by chronic-phase CML CFU-GM. Here, we studied CFU-GM in advanced-phase (accelerated and blast crisis) CML. The numbers and self-replication of CFU-GM in advanced phase were greater than in the chronic phase. Self-replication by CFU-GM from advanced phase patients was reduced by STI571 or IFN alfa to the same extent as in the chronic phase. The reduced replating ability induced by STI571 correlated with that induced by IFN alpha (r=0.73). STI571 treatment in vivo also reduced replating ability and the numbers of CFU-GM/ml of blood. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11597734&query_hl=1 ER - TY - JFULL T1 - Improved immnunophenotyping of lymphocytes in bronchoalveolar lavage fluid (BALF) by flow cytometry. A1 - Ma, W A1 - Cui, W A1 - Lin, Q J1 - Clin Chim Acta Y1 - 2001/11// VL - 313 SN - 0009-8981 SP - 133 EP - 138 N2 - BACKGROUND: The immnunophenotyping of lymphocytes in bronchoalveolar lavage fluid (BALF) is of particular importance in the differential diagnosis of interstitial lung disorders. The standard method of lymphocyte phenotyping is peroxidase-anti-peroxidase technique (PAP). However, it was time-consuming and experience-dependent. Flow cytometric (FCM) analysis of BALF lymphocytes was introduced to overcome these disadvantages. Unfortunately, when the number of cells counted was small, FCM could not distinguish lymphocytes from other cells and particles in BALF by light scattering. METHODS: We established a tri-color flow cytometric approach to phenotyping of lymphocytes in BALF. FITC-CD45/PE-CD14 antibodies were used to gate lymphocytes and exclude other contamination. Propidium iodide (PI) was introduced to distinguish lymphocytes from debris. Forty-three BALF species were tested by flow cytometer as well as peripheral blood as a control group by conventional PAP method. RESULTS: The variation of FCM (CV<1.0%) was much lower that that of PAP method (CV>9.8%). Meanwhile, we found that BALF had more clinical significance than peripheral blood in T subset analysis (p<0.01). There were characteristic changes in some lung diseases. Both CD3 and CD4 were significantly increased with decreasing CD8 in sarcoidosis (n=14). Idiopathic pulmonary fibrosis (n=16) demonstrated the reverse tendency: CD8 rose but both CD3 and CD4 dropped. As for lung cancer (n=7), CD3 was normal but the CD4/CD8 ratio declined. Tuberculosis of the lungs (n=6) showed a normal CD3, CD4 and CD8. CONCLUSIONS: The high precision and reliability of tri-color flow cytometric approach to phenotyping of lymphocytes in BALF suggested that it should be used as a routine test, especially of BALF, which was often contaminated by inorganic particles. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11694250&query_hl=1 ER - TY - JFULL T1 - Changes in sarcolemmal Ca entry and sarcoplasmic reticulum (SR) Ca content in isolated ventricular myocytes from patients with end-stage heart failure following left ventricular assist device support A1 - Terracciano, CM A1 - Koban, M A1 - Harding, SE A1 - Tansley, P A1 - Birks, EJ A1 - Yacoub, MH J1 - CIRCULATION Y1 - 2001/10/23/ VL - 104 SN - 0009-7322 SP - 480 EP - 481 ER - TY - JFULL T1 - Evidence for altered hepatic gluconeogenesis in patients with cirrhosis using in vivo 31-phosphorus magnetic resonance spectroscopy. A1 - Changani, KK A1 - Jalan, R A1 - Cox, IJ A1 - Ala-Korpela, M A1 - Bhakoo, K A1 - Taylor-Robinson, SD A1 - Bell, JD J1 - Gut Y1 - 2001/10// VL - 49 SN - 0017-5749 SP - 557 EP - 564 N2 - BACKGROUND AND AIMS: Alterations in gluconeogenesis in the diseased liver can be assessed non-invasively using magnetic resonance spectroscopy by measuring changes in phosphomonoester resonance which contains information regarding several metabolites, including the phosphorylated intermediates of the gluconeogenic pathway. METHODS: 31P magnetic resonance spectroscopy was used to determine changes in phosphomonoesters following bolus infusions of 2.8 mmol/kg L-alanine in five patients with functionally compensated cirrhosis and in five patients with functionally decompensated cirrhosis. RESULTS: Compared with six healthy volunteers, baseline phosphomonoester values were elevated by 35% (p<0.05) in the compensated cirrhosis group and by 57% (p<0.01) in the decompensated cirrhosis group. Following alanine infusion, phosphomonoesters in healthy volunteers increased by 46% from baseline values (p<0.01), in patients with compensated cirrhosis by 27% (p<0.02) but those with decompensated cirrhosis showed no increase from baseline. There was a reduction in the percentage of inorganic phosphate signal in all subjects. CONCLUSIONS: By analysing changes in phosphomonoester and inorganic phosphate resonances it is possible to discern clear metabolic differences between healthy volunteers and patients with cirrhosis of varying severity using magnetic resonance spectroscopy. Those patients with functionally decompensated cirrhosis have higher percentage baseline phosphomonoester values but the absence of phosphomonoester elevation following L-alanine infusion suggests that they are unable to mount a significant metabolic response with a progluconeogenic stimulus. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11559655&query_hl=1 ER - TY - JFULL T1 - Developmental and regional distribution of aspartoacylase in rat brain tissue. A1 - Bhakoo, KK A1 - Craig, TJ A1 - Styles, P J1 - J Neurochem Y1 - 2001/10// VL - 79 SN - 0022-3042 SP - 211 EP - 220 N2 - The function of N-acetyl-aspartate (NAA), a predominant molecule in the brain, has not yet been determined. However, NAA is commonly used as a putative marker of viable neurones. To investigate the possible function of NAA, we determined the anatomical, developmental and cellular distribution of aspartoacylase, which catalyses the hydrolysis of NAA. Levels of aspartoacylase activity were measured during postnatal development in several brain regions. The differential distribution of aspartoacylase activity in purified populations of cells derived from the rat CNS was also investigated. The developmental and anatomical distribution of aspartoacylase correlated with the maturation of white matter tracts in the rat brain. Activity increased markedly after 7 days and coincided with the time course for the onset of myelination in the rat brain. Gray matter showed little activity or developmental trend. There was a 60-fold excess in optic nerve (a white matter tract) when compared with cortex at 21 days of development. In the adult brain there was a 18-fold difference in corpus callosum compared with cortex (stripped of corpus callosum). Cellular studies demonstrated that purified cortical neurons and cerebellar granular neurones have no activity. Primary O-2A progenitor cells had moderate activity, with three-fold higher activity in immature oligodendrocyte and 13-fold increase in mature oligodendrocytes (myelinating cells of the CNS). The highest activity was seen in type-2 astrocytes (20-fold difference compared with O-2A progenitors) derived from the same source. Aspartoacylase activity increased with time in freshly isolated astrocytes, with significantly higher activity after 15 days in culture. We conclude that aspartoacylase activity in the developing postnatal brain corresponds with maturation of myelination, and that the cellular distribution is limited to glial cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11595773&query_hl=1 ER - TY - JFULL T1 - Expression of angiogenesis-related molecules in plexiform lesions in severe pulmonary hypertension: evidence for a process of disordered angiogenesis. A1 - Tuder, RM A1 - Chacon, M A1 - Alger, L A1 - Wang, J A1 - Taraseviciene-Stewart, L A1 - Kasahara, Y A1 - Cool, CD A1 - Bishop, AE A1 - Geraci, M A1 - Semenza, GL A1 - Yacoub, M A1 - Polak, JM A1 - Voelkel, NF J1 - J Pathol Y1 - 2001/10// VL - 195 SN - 0022-3417 SP - 367 EP - 374 N2 - Pulmonary arteries of patients with severe pulmonary hypertension (SPH) presenting in an idiopathic form (primary PH-PPH) or associated with congenital heart malformations or collagen vascular diseases show plexiform lesions. It is postulated that in lungs with SPH, endothelial cells in plexiform lesions express genes encoding for proteins involved in angiogenesis, in particular, vascular endothelial growth factor (VEGF) and those involved in VEGF receptor-2 (VEGFR-2) signalling. On immunohistochemistry and in situ hybridization, endothelial cells in the plexiform lesions expressed VEGF mRNA and protein and overexpressed the mRNA and protein of VEGFR-2, and the transcription factor subunits HIF-1alpha and HIF-1beta of hypoxia inducible factor, which are responsible for the hypoxia-dependent induction of VEGF. When compared with normal lungs, SPH lungs showed decreased expression of the kinases PI3 kinase and src, which, together with Akt, relay the signal transduction downstream of VEGFR-2. Because markers of angiogenesis are expressed in plexiform lesions in SPH, it is proposed that these lesions may form by a process of disordered angiogenesis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11673836&query_hl=1 ER - TY - JFULL T1 - Nitric oxide: not just a negative inotrope. A1 - Sarkar, D A1 - Vallance, P A1 - Harding, SE J1 - Eur J Heart Fail Y1 - 2001/10// VL - 3 SN - 1388-9842 SP - 527 EP - 534 N2 - Nitric oxide (NO) appears to play a role in modulating cardiac function in both health and disease. Early studies in isolated rodent cardiac myocytes demonstrated a depressant effect of NO supplied by NO donors (exogenous) as well as NO generated within myocytes (endogenous). There is increasing evidence for a functional NO generating system within the human myocardium, which appears upregulated in certain disease states. Induction of the high output nitric oxide synthase isoform (iNOS) has been demonstrated in the failing myocardium, though its functional significance remains unproven. More recently published data have contradicted the notion that NO acts solely as a negative inotrope demonstrating positive inotropy in both isolated rodent and human ventricular myocytes in response to a range of NO donors. Different NO donors have different NO release kinetics and generate a range of NO species (NO., NO+ and NO-) which may interact at a number of subcellular targets. The observed response of any cardiac preparation to an NO donor represents the net effect of activation of different effector targets and may explain the contradictory reported effects of NO. To realise the therapeutic potential of NO will require specific targeting at a subcellular level. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11595600&query_hl=1 ER - TY - JFULL T1 - Quantitative myocardial cytokine expression and activation of the apoptotic pathway in patients who require left ventricular assist devices. A1 - Birks, EJ A1 - Latif, N A1 - Owen, V A1 - Bowles, C A1 - Felkin, LE A1 - Mullen, AJ A1 - Khaghani, A A1 - Barton, PJ A1 - Polak, JM A1 - Pepper, JR A1 - Banner, NR A1 - Yacoub, MH J1 - Circulation Y1 - 2001/09/18/ VL - 104 SN - 1524-4539 SP - I233 EP - I240 N2 - BACKGROUND: Molecular mechanisms underlying the deterioration of patients undergoing LV assist device (LVAD) implantation remain poorly understood. We studied the cytokines tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta and IL-6 and the terminal stage of the apoptotic pathway in patients with decompensating heart failure who required LVAD support and compared them with patients with less severe heart failure undergoing elective heart transplantation. METHODS AND RESULTS: Myocardial and serum samples from 23 patients undergoing LVAD implantation were compared with those from 36 patients undergoing elective heart transplantation. Myocardial TNF-alpha mRNA (1.71-fold; P<0.05) and protein (3.43+/-0.19 versus 2.95+/-0.10 pg/mg protein; P<0.05) were elevated in the LVAD patients. Immunocytochemistry demonstrated TNF expression in the myocytes. Serum TNF-alpha was also elevated (12.5+/-1.9 versus 4.0+/-0.4 pg/mL; P<0.0001) in the LVAD patients. IL-6 mRNA (2.57-fold higher; P<0.005) and protein (27.83+/-9.35 versus 4.26+/-1.24 pg/mg protein; P<0.001) were higher in the LVAD candidates, as was serum IL-6 (79.3+/-23.6 versus 7.1+/-1.6 pg/mL; P<0.0001). Interleukin-1beta mRNA expression was 9.78-fold higher in the LVAD patients (P<0.001). iNOS mRNA expression was similar to that in advanced heart failure patients and was not further elevated in the LVAD patients. Levels of procaspase-9 (8.02+/-0.91 versus 6.16+/-0.43 oligodeoxynucleotide [OD] units; P<0.01), cleaved caspase-9 (10.02+/-1.0 versus 7.34+/-0.40 OD units; P<0.05), intact and spliced DFF-45 (4.58+/-0.75 versus 2.84+/-0.23 OD units; P<0.05) were raised in LVAD patients, but caspase-3 and human nuclease CPAN were not. CONCLUSIONS: Elevated TNF-alpha, IL-1beta, and IL-6 and alterations in the apoptotic pathway were found in the myocardium and elevated TNF-alpha and IL-6 in serum of deteriorating patients who required LVAD support. These occurrences may have therapeutic implications and influence the timing of LVAD insertion. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11568062&query_hl=1 ER - TY - JFULL T1 - A preliminary study on the effect of galectin-1 on cardiomyocytes A1 - Watt, DJ A1 - Goldring, K A1 - Harding, SE A1 - Fuller, S A1 - Sun, H A1 - Naunton-Morgan, N J1 - NEUROMUSCULAR DISORD Y1 - 2001/09// VL - 11 SN - 0960-8966 SP - 651 EP - 651 ER - TY - JFULL T1 - Nonequivalent nuclear location of immunoglobulin alleles in B lymphocytes. A1 - Skok, JA A1 - Brown, KE A1 - Azuara, V A1 - Caparros, ML A1 - Baxter, J A1 - Takacs, K A1 - Dillon, N A1 - Gray, D A1 - Perry, RP A1 - Merkenschlager, M A1 - Fisher, AG J1 - Nat Immunol Y1 - 2001/09// VL - 2 SN - 1529-2908 SP - 848 EP - 854 N2 - Individual B lymphocytes normally express immunoglobulin (Ig) proteins derived from single Ig heavy chain (H) and light chain (L) alleles. Allelic exclusion ensures monoallelic expression of Ig genes by each B cell to maintain single receptor specificity. Here we provide evidence that at later stages of B cell development, additional mechanisms may contribute to prioritizing expression of single IgH and IgL alleles. Fluorescent in situ hybridization analysis of primary splenic B cells isolated from normal and genetically manipulated mice showed that endogenous IgH, kappa and lambda alleles localized to different subnuclear environments after activation and had differential expression patterns. However, this differential recruitment and expression of Ig alleles was not typically seen among transformed B cell lines. These data raise the possibility that epigenetic factors help maintain the monoallelic expression of Ig. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11526401&query_hl=1 ER - TY - JFULL T1 - Simultaneous measurement of Ca2+ and cellular dynamics: combined scanning ion conductance and optical microscopy to study contracting cardiac myocytes. A1 - Shevchuk, AI A1 - Gorelik, J A1 - Harding, SE A1 - Lab, MJ A1 - Klenerman, D A1 - Korchev, YE J1 - Biophys J Y1 - 2001/09// VL - 81 SN - 0006-3495 SP - 1759 EP - 1764 N2 - We have developed a distance modulated protocol for scanning ion conductance microscopy to provide a robust and reliable distance control mechanism for imaging contracting cells. The technique can measure rapid changes in cell height from 10 nm to several micrometers, with millisecond time resolution. This has been demonstrated on the extreme case of a contracting cardiac myocyte. By combining this method with laser confocal microscopy, it was possible to simultaneously measure the nanometric motion of the cardiac myocyte, and the local calcium concentration just under the cell membrane. Despite large cellular movement, simultaneous tracking of the changes in cell height and measurement of the intracellular Ca2+ near the cell surface is possible while retaining the cell functionality. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11509385&query_hl=1 ER - TY - JFULL T1 - Differential adrenomedullin release and endothelin receptor expression in distinct subpopulations of human airway smooth-muscle cells. A1 - Upton, PD A1 - Wharton, J A1 - Davie, N A1 - Ghatei, MA A1 - Smith, DM A1 - Morrell, NW J1 - Am J Respir Cell Mol Biol Y1 - 2001/09// VL - 25 SN - 1044-1549 SP - 316 EP - 325 N2 - Although adrenomedullin (ADM) is implicated in the control of airway tone, regulation of ADM release from airway smooth-muscle cells (ASMCs) has not been explored. Preliminary experiments have indicated that human ASMC populations were heterogeneous in their rate of ADM release and expression of endothelin (ET)(A) and ET(B) receptors. We isolated these phenotypically distinct ASMCs from explants derived from the same airway segment. ASMCs possessing exclusively ET(A) receptors appeared smaller and proliferated faster than ET(A)/ET(B) isolates. Macroautoradiographic analysis confirmed the presence of both receptors in human bronchi. ADM release and messenger RNA expression was greater in ET(A)/ET(B) isolates compared with ET(A) isolates. No measurable ET release was detected from ASMCs. Exogenous ET-1 (1 to 100 nM) more potently stimulated the release of ADM from ET(A)/ET(B) compared with ET(A) isolates. In addition, ET-3 (1 to 100 nM) stimulated ADM release only from ET(A)/ET(B) isolates, implicating the ET(B) receptor in this response. Exogenous ET-1 potentiated platelet- derived growth factor-stimulated [3H]thymidine uptake in ET(A)/ ET(B) but not ET(A) isolates. ET-3 did not affect [3H]thymidine uptake in either cell type. Possession of ET(A)/ET(B) receptors is associated with higher rates of ADM release and slower proliferation, but a capacity for ET-1 stimulated DNA synthesis via ET(A) receptors. These results support a paracrine role for the regulation of ADM release predominantly via the ET(B) receptor in human ASMCs. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11588009&query_hl=1 ER - TY - JFULL T1 - Cross-talk between Stat5b and estrogen receptor-alpha and -beta in mammary epithelial cells. A1 - Björnström, L A1 - Kilic, E A1 - Norman, M A1 - Parker, MG A1 - Sjöberg, M J1 - J Mol Endocrinol Y1 - 2001/08// VL - 27 SN - 0952-5041 SP - 93 EP - 106 N2 - Both 17beta-estradiol and prolactin play important roles in the mammary gland, raising the possibility of functional cross-talk between the two signaling pathways. Here, we demonstrate that estrogen receptor-alpha (ERalpha) and -beta (ERbeta) are both able to potentiate transcription from a Stat5-responsive promoter when activated by prolactin. Potentiation was observed not only in the presence of 17beta-estradiol, but also in the presence of anti-estrogens such as tamoxifen and ICI 182,780. The magnitude of the response was dependent on cell-type: in the HC11 mouse mammary epithelial cell line ERbeta potentiates transcription efficiently whereas ERalpha showed low activity. Conversely, in COS-7 cells, both estrogen receptors were active. We show that activation domains in the N-terminus (AF-1) and the C-terminus (AF-2) of the ERs are dispensable for potentiation. The effects are dependent on the presence of an intact DNA-binding/hinge domain, which we show is capable of interacting with Stat5b in vitro and in HC11 cell extracts. We conclude that ERalpha and ERbeta act as coactivators for Stat5b through a mechanism which is independent of AF-1 and AF-2. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11463580&query_hl=1 ER - TY - JFULL T1 - Inactivation of platelet-derived growth factor-BB following modification by ADP-ribosyltransferase. A1 - Saxty, BA A1 - Yadollahi-Farsani, M A1 - Upton, PD A1 - Johnstone, SR A1 - MacDermot, J J1 - Br J Pharmacol Y1 - 2001/08// VL - 133 SN - 0007-1188 SP - 1219 EP - 1226 N2 - 1. Arginine-specific ADP-ribosyltransferase (ART1) is expressed on the surface of a number of cell types, and catalyses the transfer of ADP-ribose from NAD(+) to target proteins. We investigated whether extracellular proteins such as growth factors may serve as substrates for this enzyme, with subsequent alteration in their biological activity. Experiments were performed with rat skeletal muscle membranes and V79 Chinese hamster lung fibroblasts with doxycycline-inducible expression of human ART. 2. From a panel of growth factors, platelet-derived growth factor-BB (PDGF-BB) was found to be the best substrate for ART1, whereas the structural homologue PDGF-AA was not a substrate. Under conditions of maximum labelling 5 mol ADP-ribose was incorporated per mol of PDGF-BB. 3. Purified (ADP-ribosyl)-PDGF-BB did not stimulate a mitogenic or chemotactic response in human pulmonary smooth muscle cells, and showed a reduced capacity to bind to PDGF receptors in competition binding experiments, when compared to unmodified PDGF-BB. 4. PDGF-dependent [(3)H-methyl]-thymidine incorporation was measured in the ART1-transfected fibroblast cell line at physiological concentrations of PDGF-BB, and without addition of extracellular NAD(+). Fibroblasts expressing human ART1 at the cell surface showed reduced mitogenic responses to PDGF-BB, but not to PDGF-AA. This loss of mitogenic response in cells expressing ART1 activity was reversed by the addition of agmatine (an ART1 substrate). 5. In conclusion, we propose that PDGF-BB-dependent signalling may be regulated by post-translational modification of the growth factor by ART1 at the cell surface. This has been demonstrated in membranes of rat skeletal muscle, and the reaction confirmed in ART1-transfected fibroblasts. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11498506&query_hl=1 ER - TY - JFULL T1 - Use of suppressor mutants to probe the function of estrogen receptor-p160 coactivator interactions. A1 - Mak, HY A1 - Parker, MG J1 - Mol Cell Biol Y1 - 2001/07// VL - 21 SN - 0270-7306 SP - 4379 EP - 4390 N2 - Estrogen-dependent recruitment of coactivators by estrogen receptor alpha (ERalpha) represents a crucial step in the transcriptional activation of target genes. However, studies of the function of individual coactivators has been hindered by the presence of endogenous coactivators, many of which are potentially recruited in the presence of agonist via a common mechanism. To circumvent this problem, we have generated second-site suppressor mutations in the nuclear receptor interaction domain of p160 coactivators which rescue their binding to a transcriptionally defective ERalpha that is refractory to wild-type coactivators. Analysis of these altered-specificity receptor-coactivator combinations, in the absence of interference from endogenous coregulators, indicated that estrogen-dependent transcription from reporter genes is critically dependent on direct recruitment of a p160 coactivator in mammalian cells and that the three p160 family members serve functionally redundant roles. Furthermore, our results suggest that such a change-of-specificity mutation may act as a transposable protein-protein interaction module which provides a novel tool with which to dissect the functional roles of other nuclear receptor coregulators at the cellular level. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11390665&query_hl=1 ER - TY - JFULL T1 - Hydrocolloids in emulsions: particle size distribution and interfacial activity A1 - Huang, X A1 - Kakuda, Y A1 - Cui, W J1 - FOOD HYDROCOLLOID Y1 - 2001/07// VL - 15 SN - 0268-005X SP - 533 EP - 542 N2 - The emulsification properties of 14 hydrocolloid gums (propylene glycol alginate, gellan, carrageenan, pectin, methylcellulose, microcrystalline cellulose, gum arabic, locust bean gum, guar, xanthan, mustard, flaxseed, fenugreek, oat) were investigated. Gum dispersions were prepared in water (0.5%) and emulsified with 40% oil using a Polytron homogenizer. Emulsion stability was determined by centrifugation and storage time, surface and interfacial tension by Du Nouy ring, particle size by integrated light scattering and overall morphology by light microscopy. When compared to the other gums in this study, fenugreek produced a very stable emulsion. Fenugreek was more efficient than other gums in lowering the interfacial free energy, its emulsion was composed of very small oil droplets (70% < 1 mum) and under the light microscope appeared as uniform droplets with a narrow size distribution. (C) 2001 Published by Elsevier Science Ltd. ER - TY - JFULL T1 - Cloning of dexamethasone-induced transcript: a novel glucocorticoid-induced gene that is upregulated in emphysema. A1 - Edgar, AJ A1 - Birks, EJ A1 - Yacoub, MH A1 - Polak, JM J1 - Am J Respir Cell Mol Biol Y1 - 2001/07// VL - 25 SN - 1044-1549 SP - 119 EP - 124 N2 - To identify changes in gene expression associated with emphysema, we used differential display to compare RNA extracted from emphysematous lungs with that of unused donor tissues taken at the time of transplant. A differentially expressed sequence was identified corresponding to the 3' end of a novel human complementary DNA (cDNA) of unknown function. The human and mouse cDNA sequences were completed by 5' rapid amplification of cDNA ends. We have named it DEXI for dexamethasone-induced transcript. DEXI messenger RNA (mRNA) was upregulated 147% in emphysematous tissue compared with donor tissue. DEXI mRNA was also upregulated 230% by dexamethasone treatment of A549. The increase in expression of DEXI found in emphysema patients' tissues may be owing to their known treatment with corticosteroids. The human DEXI gene is intronless and the predicted open reading frame encodes a 95-residue acidic protein. Database searches revealed the presence of homologues only in mammals, and a human pseudogene. The protein has a predicted central transmembrane domain and a carboxy-terminal leucine zipper. The human mRNA has a single 1.3-kb transcript. We suggest that the increased expression of DEXI in emphysema may either be relevant to disease progression or be indicative of glucocorticoid responsiveness in treated patients. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11472984&query_hl=1 ER - TY - JFULL T1 - BCR-ABL and interleukin 3 promote haematopoietic cell proliferation and survival through modulation of cyclin D2 and p27Kip1 expression. A1 - Parada, Y A1 - Banerji, L A1 - Glassford, J A1 - Lea, NC A1 - Collado, M A1 - Rivas, C A1 - Lewis, JL A1 - Gordon, MY A1 - Thomas, NS A1 - Lam, EW J1 - J Biol Chem Y1 - 2001/06/29/ VL - 276 SN - 0021-9258 SP - 23572 EP - 23580 N2 - Although it is evident that BCR-ABL can rescue cytokine-deprived hematopoietic progenitor cells from cell cycle arrest and apoptosis, the exact mechanism of action of BCR/ABL and interleukin (IL)-3 to promote proliferation and survival has not been established. Using the pro-B cell line BaF3 and a BaF3 cell line stably overexpressing BCR-ABL (BaF3-p210), we investigated the proliferative signals derived from BCR-ABL and IL-3. The results indicate that both IL-3 and BCR-ABL target the expression of cyclin Ds and down-regulation of p27(Kip1) to mediate pRB-related pocket protein phosphorylation, E2F activation, and thus S phase progression. These findings were further confirmed in a BaF3 cell line (TonB.210) where the BCR-ABL expression is inducible by doxycyclin and by using the drug STI571 to inactivate BCR-ABL activity in BaF3-p210. To establish the functional significance of cyclin D2 and p27(Kip1) expression in response to IL-3 and BCR-ABL expression, we studied the effects of ectopic expression of cyclin D2 and p27(Kip1) on cell proliferation and survival. Our results demonstrate that both cyclin D2 and p27(Kip1) have a role in BaF3 cell proliferation and survival, as ectopic expression of cyclin D2 is sufficient to abolish the cell cycle arrest and apoptosis induced by IL-3 withdrawal or by BCR-ABL inactivation, while overexpression of p27(Kip1) can cause cell cycle arrest and apoptosis in the BaF3 cells. Furthermore, our data also suggest that cyclin D2 functions upstream of p27(Kip1), cyclin E, and cyclin D3, and therefore, plays an essential part in integrating the signals from IL-3 and BCR-ABL with the pRB/E2F pathway. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11323429&query_hl=1 ER - TY - JFULL T1 - Overwhelming evidence of the beneficial effects of SERCA gene transfer in heart failure A1 - del Monte, F A1 - Hajjar, RJ A1 - Harding, SE J1 - CIRC RES Y1 - 2001/06/08/ VL - 88 SN - 0009-7330 SP - E66 EP - E66 ER - TY - JFULL T1 - Conditional ablation of neurones in transgenic mice. A1 - Isles, AR A1 - Ma, D A1 - Milsom, C A1 - Skynner, MJ A1 - Cui, W A1 - Clark, J A1 - Keverne, EB A1 - Allen, ND J1 - J Neurobiol Y1 - 2001/06/05/ VL - 47 SN - 0022-3034 SP - 183 EP - 193 N2 - Conditional targeted ablation of specific cell populations in living transgenic animals is a very powerful strategy to determine cell functions in vivo. This approach would be of particular value to study the functions of distinct neuronal populations; however, the transgene of choice for conditional cell ablation studies in mice, the herpes simplex virus thymidine kinase gene, cannot be used to ablate neurones as its principal mode of action relies on cell proliferation. Here we report that expression of the E.coli nitroreductase gene (Ntr) and metabolism of the prodrug CB1954 (5-aziridin-1-yl-2-4-dinitrobenzamide) to its cytotoxic derivative can be used to conditionally and acutely ablate specific neuronal populations in vivo. As proof of principal, we have ablated olfactory and vomeronasal receptor neurones by expressing Ntr under the control of the olfactory marker protein (OMP) gene promoter. We demonstrate that following CB1954 administration, olfactory and vomeronasal receptor neurones expressing the transgene were selectively eliminated from the olfactory epithelium (OE), and projections to the olfactory bulb (OB) were lost. The functional efficacy of cell ablation was demonstrated using a highly sensitive behavioural test to show that ablated mice had lost the olfactory ability to discriminate distinct odors and were consequently rendered anosmic. Targeted expression of Ntr to specific neuronal populations using conventional transgenes, as described here, or by "knock-in" gene targeting using embryonic stem cells may be of significant value to address the functions of distinct neuronal populations in vivo. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11333400&query_hl=1 ER - TY - JFULL T1 - Superoxide dismutase in development of obliterative bronchiolitis. A1 - Salminen, U A1 - Harjula, AL A1 - Maasilta, PK A1 - Romanska, HM A1 - Bishop, AE A1 - Polak, JM J1 - Transplant Proc Y1 - 2001/06// VL - 33 SN - 0041-1345 SP - 2477 EP - 2477 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11406217&query_hl=1 ER - TY - JFULL T1 - Inducible ablation of astrocytes shows that these cells are required for neuronal survival in the adult brain. A1 - Cui, W A1 - Allen, ND A1 - Skynner, M A1 - Gusterson, B A1 - Clark, AJ J1 - Glia Y1 - 2001/06// VL - 34 SN - 0894-1491 SP - 272 EP - 282 N2 - To study the function of astrocytes in the adult brain, we have targeted the expression of E. coli nitroreductase (NTR) to the astrocytes of transgenic mice under the control of the GFAP promoter. The astrocytes expressing NTR were selectively ablated after administration of the prodrug CB1954, resulting in motor discoordination. Histological examination showed that the region most affected in the brain was the cerebellum, in which the Bergmann glia were eliminated and the granular neurons had degenerated. Specific effects were also noted on the dendrites of the Purkinje cells, and the junction between these neurons and granular layer was disrupted. Astrocyte ablation was associated with a dramatic decrease in the expression of glutamate transporters, which may account for the degeneration of granular neurons since the excitotoxic effects of glutamate result in a similar phenotype. These results provide the first evidence that astrocytes are important for the survival of neurons in the adult brain in vivo. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11360300&query_hl=1 ER - TY - JFULL T1 - Hypo-osmotic swelling-activated release of organic osmolytes in brain slices: implications for brain oedema in vivo. A1 - Bothwell, JH A1 - Rae, C A1 - Dixon, RM A1 - Styles, P A1 - Bhakoo, KK J1 - J Neurochem Y1 - 2001/06// VL - 77 SN - 0022-3042 SP - 1632 EP - 1640 N2 - A decrease in the intracellular levels of osmotically active species has invariably been seen after swelling of mammalian brain tissue preparations. The exact identity of the species, and the manner of their decrease, remain to be described. We investigated the swelling-activated decrease of organic osmolytes in rat cortical brain slices using (1)H- and (31)P-magnetic resonance spectroscopy. We found that acute hypo-osmotic shock causes decreases in the levels of a range of intracellular amino acids and amino acid derivatives, N-acetyl-aspartate, creatine, GABA, glutamate, hypotaurine, and also in the levels of the methylamines glycerol-phosphorylcholine, phosphorylcholine and choline. Incubation of cortical slices with the anion channel blockers niflumic acid and tamoxifen caused inhibition of organic osmolyte efflux, suggesting that such osmolyte efflux occurs through anion channels. Intracellular phosphocreatine was also seen to decrease during acute hypo-osmotic superfusion, although intracellular ATP remained constant. In addition, the acidification of an intracellular compartment was observed during hypo-osmotic superfusion. Our results suggest a link between brain energy reserve and brain osmoregulation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11413246&query_hl=1 ER - TY - JFULL T1 - Pantoprazole therapy in the long-term management of severe acid peptic disease: clinical efficacy, safety, serum gastrin, gastric histology, and endocrine cell studies. A1 - Bardhan, KD A1 - Cherian, P A1 - Bishop, AE A1 - Polak, JM A1 - Romanska, H A1 - Perry, MJ A1 - Rowland, A A1 - Thompson, M A1 - Morris, P A1 - Schneider, A A1 - Fischer, R A1 - Ng, W A1 - Lühmann, R A1 - McCaldin, B J1 - Am J Gastroenterol Y1 - 2001/06// VL - 96 SN - 0002-9270 SP - 1767 EP - 1776 N2 - OBJECTIVE: Pantoprazole is the third proton pump inhibitor to become available. When this study was started, there were few data on its long-term use. Our aim was to investigate this aspect and, because powerful inhibitors of acid secretion can cause hypergastrinemia and, in experimental animals, enterochromaffin-like cell hyperplasia, we also monitored serum gastrin and endocrine cell histology. METHODS: One hundred fifty patients refractory to H2-receptor antagonists, running an aggressive course or with complications, were entered into a 5-yr treatment program. We performed serial endoscopy, checked for adverse events, and laboratory values. We also monitored serum gastrin, gastric endocrine cell histology, and antral and corpus gastritis. RESULTS: This report presents results from up to 3 yr of treatment. Cumulative healing on 40-80 mg of pantoprazole was 82% at 4 wk and 92% by 12 wk. Most patients became asymptomatic within 4 wk. Remission on maintenance treatment with 40 mg (n = 111) was 85% at 12 months and 78% at 24 months. Treatment was safe; only four patients had adverse events definitely related to pantoprazole. Elevations in gastrin were modest and there were no significant changes in gastric endocrine cells. The number of enterochromaffin-like cells tended to decrease. CONCLUSION: Pantoprazole is effective, safe, and does not seem to be associated with large increases in serum gastrin or alterations in gastric endocrine cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11419827&query_hl=1 ER - TY - JFULL T1 - Rapid inhibition of the Na+-K+ pump affects Na+-Ca2+ exchanger-mediated relaxation in rabbit ventricular myocytes. A1 - Terracciano, CM J1 - J Physiol Y1 - 2001/05/15/ VL - 533 SN - 0022-3751 SP - 165 EP - 173 N2 - The direct influence of Na+-K+ pump activity on the ability of the Na+-Ca2+ exchanger to remove Ca2+ was investigated in isolated adult rabbit ventricular myocytes. Cell shortening was measured using an edge-detection system. Cytoplasmic [Ca2+] was monitored using the fluorescent indicator indo-1. Electrophysiological parameters were recorded using high-resistance microelectrodes. The Na+-K+ pump was rapidly inhibited by removal of extracellular K+ and measurements were taken almost immediately to minimise effects on other cellular compartments. Activity of the Na+-Ca2+ exchanger was monitored during release of Ca2+ from the sarcoplasmic reticulum (SR) elicited by rapid application of 15 mM caffeine. When Na+-K+ pump activity was affected by K+ removal, cell relaxation and indo-1 fluorescence decline were slowed by approximately 40 %. The charge calculated by integrating the caffeine-induced transient inward current was unchanged, suggesting that there was no difference in the SR Ca2+ content in the two conditions. However Ca2+ flux via the Na+-Ca2+ exchanger was slower when the Na+-K+ pump was inhibited. Similar experiments were performed by inhibiting the Na+-K+ pump using 0.5 mM strophanthidin. In this condition similar results to the ones observed by K+ removal were obtained, suggesting a specific role of the Na+-K+ pump in the phenomenon observed. This study suggests that the activity of the Na+-K+ pump influences Na+-Ca2+ exchanger function in the absence of changes in SR Ca2+ content. This can be explained by a slower removal of Na+ from the subsarcolemmal space. The source of the increase in subsarcolemmal [Na+] requires further investigation. However, calculations derived from modelling suggest that the Na+-Ca2+ exchanger itself could be involved. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11351025&query_hl=1 ER - TY - JFULL T1 - Gene-expression profiling of human osteoblasts following treatment with the ionic products of Bioglass 45S5 dissolution. A1 - Xynos, ID A1 - Edgar, AJ A1 - Buttery, LD A1 - Hench, LL A1 - Polak, JM J1 - J Biomed Mater Res Y1 - 2001/05// VL - 55 SN - 0021-9304 SP - 151 EP - 157 N2 - The effect of the ionic products of Bioglass 45S5 dissolution on the gene-expression profile of human osteoblasts was investigated by cDNA microarray analysis of 1,176 genes. Treatment with the ionic products of Bioglass 45S5 dissolution increased the levels of 60 transcripts twofold or more and reduced the levels of five transcripts to one-half or less than in control. Markedly up-regulated genes included RCL, a c-myc responsive growth related gene, cell cycle regulators such as G1/S specific cyclin D1, and apoptosis regulators including calpain and defender against cell death (DAD1). Other significantly up-regulated genes included the cell surface receptors CD44 and integrin beta1, and various extracellular matrix regulators including metalloproteinases-2 and -4 and their inhibitors TIMP-1 and TIMP-2. The identification of differentially expressed genes by cDNA microarray analysis has offered new insights into the mode of action of bioactive glasses and has proven to be an effective tool in evaluating their osteoproductive properties. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11255166&query_hl=1 ER - TY - JFULL T1 - Increased expression of inducible nitric oxide synthase and cyclo-oxygenase-2 in the airway epithelium of asthmatic subjects and regulation by corticosteroid treatment A1 - Redington, AE A1 - Meng, QH A1 - Springall, DR A1 - Evans, TJ A1 - Creminon, C A1 - Maclouf, J A1 - Holgate, ST A1 - Howarth, PH A1 - Polak, JM J1 - THORAX Y1 - 2001/05// VL - 56 SN - 0040-6376 SP - 351 EP - 357 N2 - Background-Nitric oxide (NO) and prostanoids are mediators of vascular and bronchial tone that are postulated to be involved in asthma. increased levels of both are found in asthmatic subjects and are synthesised by enzymes that have cytokine inducible forms: inducible NO synthase (iNOS) and cyclo-oxygenase-2 (COX-2), respectively. We hypothesised that the in vivo expression of iNOS and COX-2 in the airways would be increased in asthma, and that these cytokine inducible enzymes may represent targets for regulation by corticosteroid treatment.Methods-Bronchial biopsy specimens were obtained from three groups of subjects: atopic asthmatics treated with p, agonists alone (n=7), atopic asthmatics additionally receiving regular treatment with corticosteroids (n=8), and nonasthmatic control subjects (n=10). Expression of iNOS and COX-2 mRNA and immunoreactive protein was studied using in situ hybridisation and quantitative immunohistochemistry.Results-Immunoreactivity and the hybridisation signal for iNOS and COX-2 were mainly localised in the airway epithelium. The proportion of epithelium immunostained was significantly greater in the non-steroid treated asthmatic subjects (iNOS 8.6 (1.8)%; COX-2 26.3 (4.6)%) than either the steroid treated asthmatics (iNOS 3.4 (1.0)%, p=0.009; COX-2 13.0 (0.6)%, p=0.0015) or the non-asthmatic controls (iNOS 4.2 (0.9)%, p=0.018; COX-2 11.6 (0.6)%, p=0.0003). Similarly, the hybridisation signal was stronger in the non-steroid treated group of asthmatic subjects than in the other two groups.Conclusions-These findings highlight the potential role of the airway epithelium both as a contributor to the inflammatory process in asthma and as a target for inhaled corticosteroid treatment in this disease. ER - TY - JFULL T1 - Murine ventricular L-type Ca(2+) current is enhanced by zinterol via beta(1)-adrenoceptors, and is reduced in TG4 mice overexpressing the human beta(2)-adrenoceptor. A1 - Heubach, JF A1 - Graf, EM A1 - Molenaar, P A1 - Jäger, A A1 - Schröder, F A1 - Herzig, S A1 - Harding, SE A1 - Ravens, U J1 - Br J Pharmacol Y1 - 2001/05// VL - 133 SN - 0007-1188 SP - 73 EP - 82 N2 - 1. The functional coupling of beta(2)-adrenoceptors (beta(2)-ARs) to murine L-type Ca(2+) current (I(Ca(L))) was investigated with two different approaches. The beta(2)-AR signalling cascade was activated either with the beta(2)-AR selective agonist zinterol (myocytes from wild-type mice), or by spontaneously active, unoccupied beta(2)-ARs (myocytes from TG4 mice with 435 fold overexpression of human beta(2)-ARs). Ca(2+) and Ba(2+) currents were recorded in the whole-cell and cell-attached configuration of the patch-clamp technique, respectively. 2. Zinterol (10 microM) significantly increased I(Ca(L)) amplitude of wild-type myocytes by 19+/-5%, and this effect was markedly enhanced after inactivation of Gi-proteins with pertussis-toxin (PTX; 76+/-13% increase). However, the effect of zinterol was entirely mediated by the beta(1)-AR subtype, since it was blocked by the beta(1)-AR selective antagonist CGP 20712A (300 nM). The beta(2)-AR selective antagonist ICI 118,551 (50 nM) did not affect the response of I(Ca(L)) to zinterol. 3. In myocytes with beta(2)-AR overexpression I(Ca(L)) was not stimulated by the activated signalling cascade. On the contrary, I(Ca(L)) was lower in TG4 myocytes and a significant reduction of single-channel activity was identified as a reason for the lower whole-cell I(Ca(L)). The beta(2)-AR inverse agonist ICI 118,551 did not further decrease I(Ca(L)). PTX-treatment increased current amplitude to values found in control myocytes. 4. In conclusion, there is no evidence for beta(2)-AR mediated increases of I(Ca(L)) in wild-type mouse ventricular myocytes. Inactivation of Gi-proteins does not unmask beta(2)-AR responses to zinterol, but augments beta(1)-AR mediated increases of I(Ca(L)). In the mouse model of beta(2)-AR overexpression I(Ca(L)) is reduced due to tonic activation of Gi-proteins. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11325796&query_hl=1 ER - TY - JFULL T1 - SERCA2A overexpression decreases the incidence of aftercontractions in adult rabbit ventricular myocytes. A1 - Davia, K A1 - Bernobich, E A1 - Ranu, HK A1 - del Monte, F A1 - Terracciano, CM A1 - MacLeod, KT A1 - Adamson, DL A1 - Chaudhri, B A1 - Hajjar, RJ A1 - Harding, SE J1 - J Mol Cell Cardiol Y1 - 2001/05// VL - 33 SN - 0022-2828 SP - 1005 EP - 1015 N2 - K. Davia, E. Bernobich, H. K. Ranu, F. del Monte, C. M. N. Terracciano, K. T. MacLeod, D. L. Adamson, B. Chaudhri, R. J. Hajjar and S. E. Harding. SERCA2a Overexpression Decreases the Incidence of Aftercontractions in Adult Rabbit Ventricular Myocytes. Journal of Molecular and Cellular Cardiology (2001) 33, 1005-1015. Slow relaxation and poor contractile response to increasing stimulation frequency in failing human heart have been strongly linked to a decrease in the activity of the sarcoplasmic reticulum (SR) Ca(2+)-ATPase (SERCA2a). Restoration of SERCA2a levels using gene transfer has beneficial effects on contractile function but, like beta -adrenoceptor stimulation, could potentially produce excess SR Ca(2+), arrhythmias and cell death. We have examined the effects of SERCA2a overexpression in adult rabbit cardiac myocytes, and compared changes in relaxation with those following beta -adrenoceptor stimulation. Myocytes were infected with an adenovirus carrying both SERCA2a and green fluorescent protein (GFP) for positive identification of infected cells. Myocyte survival was significantly enhanced in the infected cultures. There was a reduction in both time-to-peak contraction and time-to-50% relaxation (R50) 48 h after infection. Time-to-90% relaxation (R90) was particularly improved (non-infected 516+/-41 ms, AD.SERCA2a-GFP 230+/-23 ms, n=7 preparations, P<0.001). There was also a decreased incidence of aftercontractions in Ad.SERCA2a-GFP infected myocytes (21+/-5%v 41+/-4% in controls, P<0.01). This contrasts with beta -adrenoceptor stimulation, which reduced R50 but prolonged R90 by 158+/-76 ms (P<0.02, n=16). At higher stimulation frequencies (2-3 Hz) contraction amplitude and SR calcium content were increased and diastolic contracture was reduced following SERCA2a overexpression. Overall, increasing levels of SERCA2a resulted in an improvement in systolic and diastolic function and a reduction in cell death and arrhythmic aftercontractions. SERCA2a overexpression therefore lacks the detrimental effects associated with some other inotropic interventions. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11343422&query_hl=1 ER - TY - JFULL T1 - The influence of INK4 proteins on growth and self-renewal kinetics of hematopoietic progenitor cells. A1 - Lewis, JL A1 - Chinswangwatanakul, W A1 - Zheng, B A1 - Marley, SB A1 - Nguyen, DX A1 - Cross, NC A1 - Banerji, L A1 - Glassford, J A1 - Thomas, NS A1 - Goldman, JM A1 - Lam, EW A1 - Gordon, MY J1 - Blood Y1 - 2001/05/01/ VL - 97 SN - 0006-4971 SP - 2604 EP - 2610 N2 - This study investigated the influence of expression of proteins of the INK4 family, particularly p16, on the growth and self-renewal kinetics of hematopoietic cells. First, retrovirus-mediated gene transfer (RMGT) was used to restore p16(INK4a) expression in the p16(INK4a)-deficient lymphoid and myeloid cell lines BV173 and K562, and it was confirmed that this inhibited their growth. Second, to sequester p16(INK4a) and related INK4 proteins, cyclin-dependent kinase 4 (CDK4) was retrovirally transduced into normal human CD34(+) bone marrow cells and then cultured in myeloid colony-forming cell (CFC) assays. The growth of CDK4-transduced colonies was more rapid; the cell-doubling time was reduced; and, upon replating, the colonies produced greater yields of secondary colonies than mock-untransduced controls. Third, colony formation was compared by marrow cells from p16(INK4a-/-) mice and wild-type mice. The results from p16(INK4a-/-) marrow were similar to those from CDK4-transduced human CFCs, in terms of growth rate and replating ability, and were partially reversed by RMGT of p16(INK4a). Lines of immature granulocytic cells were raised from 15 individual colonies grown from the marrow of p16(INK4a-/-) mice. These had a high colony-forming ability (15%) and replating efficiency (96.7%). The p16(INK4a-/-) cell lines readily became growth factor-independent upon cytokine deprivation. Taken together, these results demonstrate that loss of INK4 proteins, in particular p16(INK4a), increases the growth rate of myeloid colonies in vitro and, more importantly, confers an increased ability for clonal expansion on hematopoietic progenitor cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11313248&query_hl=1 ER - TY - JFULL T1 - Overexpression of the Na/Ca exchanger and reduced SERCa function. A1 - Terracciano, CM A1 - MacLeod, KT J1 - Cardiovasc Res Y1 - 2001/04// VL - 50 SN - 0008-6363 SP - 167 EP - 169 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11345943&query_hl=1 ER - TY - JFULL T1 - Human CD1d-glycolipid tetramers generated by in vitro oxidative refolding chromatography A1 - Karadimitris, A A1 - Gadola, S A1 - Altamirano, M A1 - Brown, D A1 - Woolfson, A A1 - Klenerman, P A1 - Chen, JL A1 - Koezuka, Y A1 - Roberts, IAG A1 - Price, DA A1 - Dusheiko, G A1 - Milstein, C A1 - Fersht, A A1 - Luzzatto, L A1 - Cerundolo, V J1 - P NATL ACAD SCI USA Y1 - 2001/03/13/ VL - 98 SN - 0027-8424 SP - 3294 EP - 3298 N2 - CD1 molecules are specialized in presenting lipids to T lymphocytes, but identification and isolation of CD1-restricted lipid-specific T cells has been hampered by the lack of reliable and sensitive techniques. We here report the construction of CD1d-glycolipid tetramers from fully denatured human CD1d molecules by using the technique of oxidative refolding chromatography. We demonstrate that chaperone- and foldase-assisted refolding of denatured CD1d molecules and beta (2)-microglobulin in the presence of synthetic lipids is a rapid method for the generation of functional and specific CD1d tetramers, which unlike previously published protocols ensures isolation of CD1d tetramers loaded with a single lipid species. The use of human CD1d-alpha -galactosylceramide tetramers for ex vivo staining of peripheral blood lymphocytes and intrahepatic T cells from patients with viral liver cirrhosis allowed for the first time simultaneous analysis of frequency and specificity of natural killer T cells in human clinical samples, Application of this protocol to other members of the CD1 family will provide powerful tools to investigate lipid-specific T cell immune responses in health and in disease. ER - TY - JFULL T1 - Core LXXLL motif sequences in CREB-binding protein, SRC1, and RIP140 define affinity and selectivity for steroid and retinoid receptors. A1 - Heery, DM A1 - Hoare, S A1 - Hussain, S A1 - Parker, MG A1 - Sheppard, H J1 - J Biol Chem Y1 - 2001/03/02/ VL - 276 SN - 0021-9258 SP - 6695 EP - 6702 N2 - An alpha-helical motif containing the sequence LXXLL is required for the ligand-dependent binding of transcriptional co-activators to nuclear receptors. By using a peptide inhibition assay, we have defined the minimal "core" LXXLL motif as an 8-amino acid sequence spanning positions -2 to +6 relative to the primary conserved leucine residue. In yeast two-hybrid assays, core LXXLL motif sequences derived from steroid receptor co-activator (SRC1), the 140-kDa receptor interacting protein (RIP140), and CREB-binding protein (CBP) displayed differences in selectivity and affinity for nuclear receptor ligand binding domains. Although core LXXLL motifs from SRC1 and RIP140 mediated strong interactions with steroid and retinoid receptors, three LXXLL motifs present in the global co-activator CBP were found to have very weak affinity for these proteins. Core motifs with high affinity for steroid and retinoid receptors were generally found to contain a hydrophobic residue at position -1 relative to the first conserved leucine and a nonhydrophobic residue at position +2. Our results indicate that variant residues in LXXLL core motifs influence the affinity and selectivity of co-activators for nuclear receptors. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11078741&query_hl=1 ER - TY - JFULL T1 - Worldwide trends in mortality rates for hepatobiliary and pancreatic tumours, 1979-1997 A1 - Khan, SA A1 - Taylor-Robinson, SD A1 - Toledano, MB A1 - Elliott, P A1 - Thomas, HC J1 - GUT Y1 - 2001/03// VL - 48 SN - 0017-5749 SP - A103 EP - A103 ER - TY - JFULL T1 - Biochemical characterisation and gene expression profiling of human trabecular bone derived osteoblasts A1 - Xynos, ID A1 - Edgar, AJ A1 - Ramachandran, M A1 - Buttery, LDK A1 - Hench, LL A1 - Polak, JM J1 - J PATHOL Y1 - 2001/03// VL - 193 SN - 0022-3417 SP - 31A EP - 31A ER - TY - JFULL T1 - Differentiation of osteoblasts and in vitro bone formation from murine embryonic stem cells. A1 - Buttery, LD A1 - Bourne, S A1 - Xynos, JD A1 - Wood, H A1 - Hughes, FJ A1 - Hughes, SP A1 - Episkopou, V A1 - Polak, JM J1 - Tissue Eng Y1 - 2001/02// VL - 7 SN - 1076-3279 SP - 89 EP - 99 N2 - Pluripotent embryonic stem (ES) cells have the potential to differentiate to all fetal and adult cell types and might represent a useful cell source for tissue engineering and repair. Here we show that differentiation of ES cells toward the osteoblast lineage can be enhanced by supplementing serum-containing media with ascorbic acid, beta-glycerophosphate, and/or dexamethasone/retinoic acid or by co-culture with fetal murine osteoblasts. ES cell differentiation into osteoblasts was characterized by the formation of discrete mineralized bone nodules that consisted of 50-100 cells within an extracellular matrix of collagen-1 and osteocalcin. Dexamethasone in combination with ascorbic acid and beta-glycerophosphate induced the greatest number of bone nodules and was dependent on time of stimulation with a sevenfold increase when added to ES cultures after, but not before, 14 days. Co-culture with fetal osteoblasts also provided a potent stimulus for osteogenic differentiation inducing a fivefold increase in nodule number relative to ES cells cultured alone. These data demonstrate the application of a quantitative assay for the derivation of osteoblast lineage progenitors from pluripotent ES cells. This could be applied to obtain purified osteoblasts to analyze mechanisms of osteogenesis and for use of ES cells in skeletal tissue repair. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11224927&query_hl=1 ER - TY - JFULL T1 - Adrenomedullin expression and growth inhibitory effects in distinct pulmonary artery smooth muscle cell subpopulations. A1 - Upton, PD A1 - Wharton, J A1 - Coppock, H A1 - Davie, N A1 - Yang, X A1 - Yacoub, MH A1 - Ghatei, MA A1 - Polak, JM A1 - Bloom, SR A1 - Smith, DM A1 - Morrell, NW J1 - Am J Respir Cell Mol Biol Y1 - 2001/02// VL - 24 SN - 1044-1549 SP - 170 EP - 178 N2 - The vasodilator peptide adrenomedullin is elevated in patients with pulmonary hypertension and has been implicated in the inhibition of vascular remodeling. We questioned whether adrenomedullin is released by human pulmonary artery smooth muscle cells (PASMCs) and inhibits PASMC growth and release of endothelin, a known smooth muscle cell mitogen. The majority of PASMCs isolated from proximal pulmonary arteries and all PASMCs from distal pulmonary arteries released adrenomedullin, although at differing rates (mean, 177 +/- 28 and 62 +/- 11 fmol/10(5) cells/24 h, respectively). These cells were designated ADM+. However, some proximal PASMC isolates did not release adrenomedullin, designated ADM-. Northern blot analysis confirmed adrenomedullin expression in proximal ADM+ but not ADM- isolates. ADM- and distal ADM+ PASMCs proliferated faster in serum than did proximal ADM+ cells. Adrenomedullin potently and dose-dependently (mean EC(50) = 2.2 +/- 0.5 nM) increased intracellular cyclic adenosine monophosphate (cAMP) in ADM- isolates via specific adrenomedullin receptors. In contrast, both adrenomedullin and calcitonin gene-related peptide modestly elevated cAMP in 50% of ADM+ isolates. Adrenomedullin dose-dependently inhibited platelet-derived growth factor-stimulated [3H]thymidine incorporation and endothelin release in ADM- cells but did not affect [3H]thymidine uptake in ADM+ isolates. We conclude that distinct subpopulations of human PASMCs release and respond to adrenomedullin. The heterogeneity of adrenomedullin release and the inhibition of PASMC DNA synthesis and endothelin release suggest that adrenomedullin may function as a paracrine mediator in the inhibition of pulmonary vascular remodeling. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11159051&query_hl=1 ER - TY - JFULL T1 - Peripheral blood progenitor cell mobilisation alters myeloid, but not erythroid, progenitor cell self-renewal kinetics. A1 - Marley, SB A1 - Lewis, JL A1 - Zheng, B A1 - Davidson, RJ A1 - Davis, JG A1 - McDonald, C A1 - Alenzi, FQ A1 - Goldman, JM A1 - Gordon, MY J1 - Bone Marrow Transplant Y1 - 2001/02// VL - 27 SN - 0268-3369 SP - 241 EP - 248 N2 - Transplantation of progenitor cells which have been mobilised into the bloodstream (PBPC) following the administration of G-CSF results in more rapid neutrophil recovery than transplantation of bone marrow (BM). The reasons for the accelerated neutrophil engraftment are not clear, but would be explained by increased self-replication of myeloid progenitor cells (CFU-GM). We have used a CFU-GM replating assay to investigate myeloid progenitor self-replication, and quantification of subcolony formation during erythroid burst formation to quantify erythroid progenitor self-renewal. Secondary colony formation by CFU-GM, grown from PBPC and then replated was increased compared with secondary colony formation by BM CFU-GM (P = 0.0001); erythroid subcolony formation was not altered. There was no difference between the replating abilities of PBPC CFU-GM derived from allogeneic donors (normal individuals) and autologous donors (patients with malignant disease) although differences were found between subgroups of autologous donors. The increased replication of PBPC could not be accounted for by a reduction in progenitor cell apoptosis; PBPC CFU-GM contained slightly fewer apoptotic CD34+ cells than BM CFU-GM. The increased replication by PBPC CFU-GM was reversible because it declined when CFU-GM colonies were passaged through three sequential CFU-GM replating cycles. This decline in self-replication was more rapid than the decline seen in replated BM CFU-GM. The self-replication of PBPC CFU-GM, and subcolony formation by BFU-E could be further enhanced by exposure to cytokines in vitro. We conclude that mobilisation alters the replication kinetics of myeloid, but not of erythroid, progenitor cells, that mobilisation-induced events are of limited duration and that in vitro exposure to cytokines may modify PBPC progenitor cell kinetics. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11277171&query_hl=1 ER - TY - JFULL T1 - Strain-specific TCR repertoire selection of IL-4-producing Thy-1 dull gamma delta thymocytes. A1 - Azuara, V A1 - Grigoriadou, K A1 - Lembezat, MP A1 - Nagler-Anderson, C A1 - Pereira, P J1 - Eur J Immunol Y1 - 2001/01// VL - 31 SN - 0014-2980 SP - 205 EP - 214 N2 - Thy-1 dull gamma delta thymocytes constitute an unusual subset of mature TCR gamma delta cells which share with NK T cells the expression of cell surface markers usually associated with activated or memory cells and the simultaneous production of high levels of IL-4 and IFN-gamma upon activation. In DBA / 2 mice, Thy-1 dull gamma delta thymocytes express a restricted repertoire of TCR that are composed of the V1 gene product mainly associated with V6.4 chains exhibiting very limited junctional sequence diversity. In this study we have characterized this gamma delta T cell population in different strains of mice and show that Thy-1 dull gamma delta thymocytes are present in every strain tested, albeit at different frequencies. Moreover IL-4 production by gamma delta thymocytes is mainly confined to the Thy-1 dull population in every strain tested. Finally, the repertoire of TCR expressed by Thy-1 dull gamma delta thymocytes varies in different strain of mice, although a biased expression of Vgamma1 and Vdelta6 chains was observed in all strains studied. However, the extent of junctional diversity of the V1 and V6 chains expressed by Thy-1 dull gamma delta thymocytes varied from oligoclonal in DBA/2 mice to polyclonal in FVB/N mice. Thy-1 dull gamma delta thymocytes from mouse strains such as C3H/HeJ and BALB/c contain cells with diverse Vdelta6(D)Jdelta junctions together with cells with relatively homogeneous Vdelta6(D)Jdelta junctions, similar to those found in DBA/2. Thus, the Thy-1 dull gamma delta population appears to contain two subsets of cells which differ in the diversity of their TCR. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11265636&query_hl=1 ER - TY - JFULL T1 - Recent Hubble space telescope results on X-ray novae A1 - Hynes, RI A1 - Haswell, CA A1 - Chaty, S A1 - Shrader, CR A1 - Cui, W A1 - Mauche, CW J1 - ASTROPHYS SPACE SCI Y1 - 2001/// VL - 276 SN - 0004-640X SP - 61 EP - 62 N2 - RXTE has recently discovered two new optically bright black hole candidates (BHCs): XTE J1859+226 and XTE J1118+490. We observed both with HST/STIS, the first outburst observations of X-ray novae with this instrument. we followed broad band spectral evolution, detected correlated X-ray/UV rapid variability and performed high resolution UV spectroscopy. We present a summary of initial results on the spectral energy distributions observed. ER - TY - JFULL T1 - Collagen IV synthesis is restricted to the enteroendocrine pathway during multilineage differentiation of human colorectal epithelial stem cells A1 - Kirkland, SC A1 - Henderson K J1 - Journal of Cell Science Y1 - 2001/// VL - 114 SP - 2055 EP - 2064 ER - TY - JFULL T1 - Effect of adhesion on inducible nitric oxide synthase (iNOS) production in purified human neutrophils. A1 - Webb, JL A1 - Polak, JM A1 - Evans, TJ J1 - Clin Exp Immunol Y1 - 2001/01// VL - 123 SN - 0009-9104 SP - 42 EP - 48 N2 - The production of nitric oxide (NO) within neutrophils is an important element of the innate immune response. We have previously shown that cytokines (IL-1alpha, tumour necrosis factor-alpha and interferon-gamma) induce human neutrophils in buffy coat preparations to produce iNOS. In order to define better the exact requirements for iNOS production within human neutrophils, we have studied the conditions needed for the production of iNOS in purified neutrophils. In contrast to buffy coat preparations, purified neutrophils in suspension did not produce an increase in iNOS following addition of cytokines. However, when purified neutrophils were allowed to adhere to glass surfaces either uncoated or coated with fetal calf serum (FCS), plasma, fibronectin or laminin, there was an increase in the percentage of iNOS-positive cells. The addition of cytokines during adhesion of these cells increased this proportion further. This was most marked for glass alone and FCS-coated glass on which the proportion of iNOS-positive cells increased to 22.7% and 35.5%, respectively, a significant increase compared with cytokine-treated neutrophils in suspension. Neither transmigration through activated endothelial monolayers nor the addition of soluble intercellular adhesion molecule-1 to purified neutrophil suspensions increased the percentage of iNOS-positive cells following cytokine stimulation. Adhesion of neutrophils to surfaces coated with IgG or complement also failed to increase cytokine-induced iNOS production. We conclude that iNOS production in human neutrophils requires not only cytokine stimulation, but also additional stimuli from adhesion to a surface. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11167996&query_hl=1 ER - TY - JFULL T1 - Overexpression of the Na(+)/Ca(2+) exchanger and inhibition of the sarcoplasmic reticulum Ca(2+)-ATPase in ventricular myocytes from transgenic mice. A1 - Terracciano, CM A1 - Philipson, KD A1 - MacLeod, KT J1 - Cardiovasc Res Y1 - 2001/01// VL - 49 SN - 0008-6363 SP - 38 EP - 47 N2 - BACKGROUND: Myocytes from failing hearts produce slower and smaller Ca(2+) transients associated with reduction in expression of sarcoplasmic reticulum (SR) Ca(2+) ATPase and an overexpression of Na(+)/Ca(2+) exchanger. Since the physiological role of both these proteins is competing for, and removing, Ca(2+) from the cytoplasm, overexpression of the exchanger may compensate for less effective SR Ca(2+) uptake. This study demonstrates this compensatory effect and provides a quantitative description of the results. METHODS: Ventricular myocytes from transgenic mice overexpressing the Na(+)/Ca(2+) exchanger (TR) and nontransgenic littermates (NON) were used. Cell shortening, cytoplasmic [Ca] (using indo-1 AM) and electrophysiological parameters were monitored. RESULTS: TR myocytes displayed faster Ca(2+) transients and twitches compared with NON myocytes. Superfusion with thapsigargin prolonged the time-course of Ca(2+) transients of TR myocytes until these were equal to the ones measured in NON myocytes. The amount of SR Ca(2+)-ATPase (SERCA) inhibition needed to obtain such transients was calculated as a function of V(max) for the Ca(2+) flux via SERCA and found to be 28%. In TR myocytes V(max) for the Ca(2+) flux via Na(+)/Ca(2+)exchange was 240% of NON myocytes. When Ca(2+) transients in TR myocytes were slowed by thapsigargin to similar values to the ones recorded in NON myocytes, SR Ca(2+) content was also correspondingly reduced. CONCLUSIONS: The results suggest that in pathophysiological conditions where there is a reduction in SERCA function, overexpression of Na(+)/Ca(2+) exchanger can compensate and allow normal Ca(2+) homeostasis to be maintained. In mouse ventricular myocytes a 2.4-fold increase in Na(+)/Ca(2+) exchange activity compensates for a reduction in SERCA function by 28% so maintaining the duration of the Ca(2+) transient. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11121794&query_hl=1 ER - TY - JFULL T1 - Deconvolving the nucleus of centaurus using a Chandra PSF library A1 - Karovska, M A1 - Aldcroft, T A1 - Elvis, MS A1 - Evans, IN A1 - Fabbiano, G A1 - Gaetz, TJ A1 - Isobe, T A1 - Jerius, D A1 - Jones, C A1 - Juda, M A1 - Kenter, A A1 - Kim, DW A1 - Kraft, RP A1 - Murray, SS A1 - Prestwich, AH A1 - Primini, F A1 - Schwartz, DA A1 - Cui, W A1 - Schreier, EJ J1 - IAU SYMP Y1 - 2001/// SN - 0074-1809 SP - 66 EP - 69 N2 - We describe preliminary results from our study of multi-scale structures in Centaurus A (NGC 5128) obtained using the Chandra X-ray Observatory HRC-I observations. The high-angular resolution Chandra images reveal X-ray multi-scale structures in this object with unprecedented detail and clarity. The region surrounding the Cen A nucleus, believed to be associated with