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| Effects of stem cell therapy on left ventricular remodeling after acute myocardial infarction: a meta-analysis.
June 1, 2010 at 2:45 PM
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| | Effects of stem cell therapy on left ventricular remodeling after acute myocardial infarction: a meta-analysis. Clin Cardiol. 2010 May;33(5):296-302 Authors: Sun L, Zhang T, Lan X, Du G OBJECTIVES: The objective was to perform a meta-analysis of clinical trials that investigated the effects of stem cell therapy on left ventricular remodeling in patients after acute myocardial infarction (AMI). BACKGROUND: Intracoronary injection of stem cells in the acute phase of myocardial infarction has been proposed to replace the cardiomyocytes lost and to prevent deleterious pathological remodeling after myocardial infarction. Previously published trials have investigated the effects of cell therapy on left ventricular (LV) remodeling in AMI patients. However, the sample size of these studies is small and the conclusions are inconsistent. METHODS: Trials were identified in the Cochrane Library, EMBASE, and PubMed databases, reviews, and reference lists of relevant articles. The weighted mean difference (WMD) was calculated for net changes in LV end-diastolic and end-systolic volumes (LVEDV and LVESV) by using fixed-effect models. RESULTS: A total of 11 tria! ls (13 comparisons) with 832 participants evaluated the association between stem cell therapy and changes in LVEDV. Compared with the control group, stem cell therapy did not influence the LVEDV changes from baseline to follow-up (WMD: - 1.76 mL, 95% confidence interval [CI]: - 4.61 to 1.08 mL, P = 0.233). A total of 9 trials (11 comparisons) with 797 participants evaluated the association between stem cell therapy and changes in LVESV. Compared with the control group, patients in the cell therapy group had a significantly greater reduction in LVESV from baseline to follow-up (WMD: - 5.08 mL, 95% CI: - 7.80 to - 2.37 mL, P < 0.001). CONCLUSION: This meta-analysis suggests that cell therapy improves left ventricular contractility, but has no effect on LV remodeling. Copyright (c) 2010 Wiley Periodicals, Inc. PMID: 20513068 [PubMed - in process] | | |
| Preapoptotic cell stress response of primary hepatocytes.
June 1, 2010 at 2:45 PM
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| | Preapoptotic cell stress response of primary hepatocytes. Hepatology. 2010 Jun;51(6):2140-51 Authors: Nipic D, Pirc A, Banic B, Suput D, Milisav I Primary hepatocytes are an important in vitro model for studying metabolism in man. Caspase-9 and Bcl-2-associated X protein (Bax) are regulators of the apoptotic pathway. Here we report on the translocation of procaspase-9 and Bax from cytoplasm to nuclei as well as on dispersion of mitochondria; these processes occur after isolation of primary hepatocytes. The observed changes appear similar to those at the beginning of apoptosis; however, the isolated hepatocytes are not apoptotic for the following reasons: (1) cells have a normal morphology and function; (2) the mitochondria are energized; (3) there is no apoptosis unless it is induced by, e.g., staurosporine or nodularin. Staurosporine does not trigger apoptosis through activation of caspase-9, as its activity is detected later than that of caspase-3. We propose that the translocation of procaspase-9 and Bax into the nuclei reduces the ability to trigger apoptosis through the intrinsic apoptotic pathway. The ! shifts of procaspase-9 and Bax are reversible in the absence of the apoptotic trigger; the spontaneous reversion was confirmed experimentally for procaspase-9, whereas Bax shifted from the nuclei to the cytosol and mitochondria after the initiation of apoptosis. To distinguish this process from apoptosis, we call it preapoptotic cell stress response. It shares some features with apoptosis; however, it is reversible and apoptosis has to be induced in addition to this process. Conclusion: Knowledge on preapoptotic cell stress response is important for assessing the quality of the cells used in cell therapies, in regenerative medicine, and of those used for modeling metabolic processes. Hepatology 2010;51:2140-2151. PMID: 20513000 [PubMed - in process] | | |
| Development of Antibody-Carrying Microbubbles Based on Clinically Available Ultrasound Contrast Agent for Targeted Molecular Imaging: A Preliminary Chemical Study.
June 1, 2010 at 2:45 PM
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| | Development of Antibody-Carrying Microbubbles Based on Clinically Available Ultrasound Contrast Agent for Targeted Molecular Imaging: A Preliminary Chemical Study. Mol Imaging Biol. 2010 May 29; Authors: Otani K, Yamahara K PURPOSE: The purpose of the study was to examine the feasibility of an antibody-carrying targeted-bubble preparation using clinically available phosphatidylserine (PS)-containing perfluorobutane-filled microbubbles for molecular ultrasound imaging. PROCEDURES: Firstly, we examined whether PS on the surface of perfluorobutane-filled microbubbles could be detected by means of flow cytometry (fluorescence activated cell sorting (FACS)) using annexin V. After conjugation with fluorescein isothiocyanate (FITC)-labeled annexin V (up to 50 muL) for 15 min on ice, microbubbles were assessed using a FACSCalibur. Secondly, we examined whether phycoerythrin (PE)-labeled streptavidin could be attached onto PS-containing perfluorobutane-filled microbubbles through the intermediacy of biotinylated annexin V. Microbubbles conjugated with biotinylated annexin V were incubated with PE-streptavidin for 30 min on ice, then FACS analysis was performed. Finally, we examined whether at! tachment of biotinylated IgG onto PS-containing perfluorobutane-filled microbubbles could be accomplished using biotinylated annexin V and avidin-biotin binding. Microbubbles with avidin-biotin complexes were incubated with Alexa488-labeled biotinylated IgG for 30 min on ice. RESULTS: FITC-positive microbubbles could be detected after conjugation with FITC-annexin V. Additionally, the mean fluorescence intensity of Sonazoid bubbles increased in a dose-dependent manner (0 muL, 3.3 vs. 50 muL, 617.1). The PE signal of microbubbles in the presence of biotinylated annexin V was higher than that in the absence of biotinylated annexin V (mean fluorescence intensity, 327.1 vs. 14.8). Significant amplification of the Alexa488-signal was accomplished through the intermediation of biotinylated annexin V and streptavidin. CONCLUSIONS: Our results support the feasibility of an antibody-carrying targeted-bubble preparation based on clinically available PS-containing perfluorobutane-fill! ed microbubbles. Although further study is needed, this techni! que coul d be applicable for in vivo molecular ultrasound imaging. PMID: 20512420 [PubMed - as supplied by publisher] | | |
| Lipid peroxidation product 4-hydroxynonenal as factor of oxidative homeostasis supporting bone regeneration with bioactive glasses.
June 1, 2010 at 2:45 PM
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| | Lipid peroxidation product 4-hydroxynonenal as factor of oxidative homeostasis supporting bone regeneration with bioactive glasses. Acta Biochim Pol. 2010 May 31; Authors: Mrakovcic L, Wildburger R, Jaganjac M, Cindric M, Cipak A, Borovic Sunjic S, Waeg G, Mogus Milankovic A, Zarkovic N Bone regeneration is a process of vital importance since fractures of long bones and large joints have a highly deleterious impact on both, individuals and society. Numerous attempts have been undertaken to alleviate this severe medical and social problem by development of novel bioactive materials, among which bioactive glass is the most attractive because of its osteoconductive and osteostimulative properties. Since lipid peroxidation is an important component of systematic stress response in patients with traumatic brain injuries and bone fractures, studies have been undertaken of the molecular mechanisms of the involvement of 4-hydroxynonenal (HNE), an end product of lipid peroxidation, in cellular growth regulation. We found that HNE generated in bone cells grown in vitro on the surfaces of bioactive glasses 45S5 and 13-93. This raises an interesting possibility of combined action of HNE and ionic bioglass dissolution products in enhanced osteogenesis probabl! y through a mitogen-activated protein kinase (MAPK) pathway. While the proposed mechanism still has to be elucidated, the finding of HNE generation on bioglass offers a new interpretation of the osteoinducting mechanisms of bioglass and suggests the possibility of tissue engineering based on manipulations of oxidative homeostasis. PMID: 20512168 [PubMed - as supplied by publisher] | | |
| Hey2 acts upstream of Notch in hematopoietic stem cell specification in zebrafish embryos.
June 1, 2010 at 2:45 PM
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| | Hey2 acts upstream of Notch in hematopoietic stem cell specification in zebrafish embryos. Blood. 2010 May 28; Authors: Rowlinson JM, Gering M Haematopoietic stem cells (HSCs) are essential for homeostasis and injury-induced regeneration of the vertebrate blood system. Although HSC transplantations constitute the most common type of stem cell therapy applied in the clinic we know relatively little about the molecular programming of HSCs during vertebrate embryogenesis. In vertebrate embryos, HSCs form in close association with the ventral wall of the dorsal aorta (DA). We have shown previously that in zebrafish HSC formation depends on the presence of a signalling cascade that involves Hedgehog, vascular endothelial growth factor (Vegf) and Notch signalling. Here, we reveal that Hey2, a hairy and enhancer-of-split related basic helix-loop-helix transcription factor often believed to act downstream of Notch, is also required for HSC formation. In DA progenitors, Hey2 expression is induced downstream of cloche and the transcription factor Scl/Tal1, and is maintained by Hedgehog and Vegf signalling. While k! nock-down of Hey2 expression results in a loss of Notch receptor expression in the DA angioblasts, activation of Notch signalling in hey2 morphants rescues HSC formation in zebrafish embryos. These results establish an essential role for Hey2 upstream of Notch in HSC formation. PMID: 20511544 [PubMed - as supplied by publisher] | | |
| Inhibition of neointimal formation and hyperplasia in vein grafts by external stent/sheath.
June 1, 2010 at 2:45 PM
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| | Inhibition of neointimal formation and hyperplasia in vein grafts by external stent/sheath. Vasc Med. 2010 May 28; Authors: Desai M, Mirzay-Razzaz J, Delft DV, Sarkar S, Hamilton G, Seifalian AM Synthetic and to a lesser extent vein graft failure is still a major problem in the treatment of peripheral arterial disease, with neointimal hyperplasia being the main cause for graft occlusion in the medium and long term. This review aims to establish the current status of external stents or sheaths in the prevention of intimal hyperplasia in small diameter (< 6 mm) vein grafts. PMID: 20511293 [PubMed - as supplied by publisher] | | |
| Artificial Engineering of Secondary Lymphoid Organs.
June 1, 2010 at 2:45 PM
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| | Artificial Engineering of Secondary Lymphoid Organs. Adv Immunol. 2010;105C:131-157 Authors: Tan JK, Watanabe T Secondary lymphoid organs such as spleen and lymph nodes are highly organized immune structures essential for the initiation of immune responses. They display distinct B cell and T cell compartments associated with specific stromal follicular dendritic cells and fibroblastic reticular cells, respectively. Interweaved through the parenchyma is a conduit system that distributes small antigens and chemokines directly to B and T cell zones. While most structural aspects between lymph nodes and spleen are common, the entry of lymphocytes, antigen-presenting cells, and antigen into lymphoid tissues is regulated differently, reflecting the specialized functions of each organ in filtering either lymph or blood. The overall organization of lymphoid tissue is vital for effective antigen screening and recognition, and is a feature which artificially constructed lymphoid organoids endeavor to replicate. Synthesis of artificial lymphoid tissues is an emerging field that aims t! o provide therapeutic application for the treatment of severe infection, cancer, and age-related involution of secondary lymphoid tissues. The development of murine artificial lymphoid tissues has benefited greatly from an understanding of organogenesis of lymphoid organs, which has delineated cellular and molecular elements essential for the recruitment and organization of lymphocytes into lymphoid structures. Here, the field of artificial lymphoid tissue engineering is considered including elements of lymphoid structure and development relevant to organoid synthesis. PMID: 20510732 [PubMed - as supplied by publisher] | | |
| Lunatic fringe potentiates Notch signaling in the developing brain.
June 1, 2010 at 2:45 PM
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| | Lunatic fringe potentiates Notch signaling in the developing brain. Mol Cell Neurosci. 2010 May 24; Authors: Kato TM, Kawaguchi A, Kosodo Y, Niwa H, Matsuzaki F Notch signaling is essential for the self-renewal of mammalian neural progenitor cells. A variety of mechanisms modulate Notch signaling to balance the self-renewal and differentiation of progenitor cells. Fringe is a major Notch regulator and promotes or suppresses Notch signaling, depending on the Notch ligands. In the developing brain, Lunatic fringe (Lfng) is expressed in self-renewing progenitors, but its roles are unknown. In this study, in vivo mosaic analyses using in utero electroporation were developed to investigate the roles of Lfng in neural progenitor cells. We found that Lfng potentiates Notch signaling cell-autonomously. Its depletion did not affect the balance between neuronally committed cells and self-renewing progenitors, however, irrespective of the cell density of Lfng-depleted cells, and caused no obvious defects in brain development. In vivo overexpression experiments with Notch ligands suggest that Lfng strongly augments Notch signaling me! diated by Delta-like 1 but not Jagged 1. PMID: 20510365 [PubMed - as supplied by publisher] | | |
| Stem cells and regenerative medicine in urology, part 1: General concepts, kidney, testis and urinary incontinence.
June 1, 2010 at 2:45 PM
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| | Stem cells and regenerative medicine in urology, part 1: General concepts, kidney, testis and urinary incontinence. Actas Urol Esp. 2010 Jun;34(6):510-515 Authors: Pastor-Navarro T, Moratalla-Charcos LM, Bermell-Marco L, Beamud-Cortés M, Osca-GarcÃa JM, Gil-Salom M INTRODUCTION: Progress in stem cell study and tissue engineering reached during the last times proves that this may be one of the most promising research fields in the future. Most urological diseases could profit from the development of disciplines such as regenerative medicine as, up to now, there have been encouraging results in this subject. MATERIAL AND METHODS: We performed an electronic research through the Pubmed database, of both original and review publications, with the following search criteria: stem cells urology, kidney stem cells, testis stem cells, urinary sphincter, cell therapy urology, tissue engeneering urology y regenerative medicine urology.RESULTS: We reviewed 33 articles published up to January 2010, trying to summarize the most relevant findings within the last years, the clinical applications and the point we have come to this day. CONCLUSION: Cell therapy and regenerative medicine are showing themselves to be one of the most promising fi! elds within urological basic investigation in the last years. However, there is much work to be done yet, to make the advances reached in basic research be applicable to the clinic. PMID: 20510113 [PubMed - as supplied by publisher] | | |
| Induction of neuro-protective/regenerative genes in stem cells infiltrating post-ischemic brain tissue.
June 1, 2010 at 2:45 PM
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| | Induction of neuro-protective/regenerative genes in stem cells infiltrating post-ischemic brain tissue. Exp Transl Stroke Med. 2010 May 28;2(1):11 Authors: Yilmaz G, Alexander JS, Erkuran Yilmaz C, Granger DN ABSTRACT: BACKGROUND: Although the therapeutic potential of bone marrow-derived stromal stem cells (BMSC) has been demonstrated in different experimental models of ischemic stroke, it remains unclear how stem cells induce neuroprotection following stroke. In this study, we describe a novel method for isolating BMSC that infiltrate postischemic brain tissue and use this method to identify the genes that are persistently activated or depressed in BMSC that infiltrate brain tissue following ischemic stroke. Methods- Ischemic strokes were induced in C57BL/6 mice by middle cerebral artery occlusion for 1 h, followed by reperfusion. BMSC were isolated from H-2Kb-tsA58 (immortomouse TM) mice, and were administered (i.v.) 24 h after reperfusion. At the peak of therapeutic improvement (14 days after the ischemic insult), infarcted brain tissue was isolated, and the BMSC were isolated by culturing at 33 degrees Celcius. Microarray analysis and RT-PCR were performed to compa! re differential gene expression between naive and infiltrating BMSC populations. Results- Z-scoring revealed dramatic differences in the expression of extracellular genes between naive and infiltrating BMSC. Pair-wise analysis detected 80 extracellular factor genes that were up-regulated ( 2 fold, P<0.05, Benjamini-Hochberg correction) between naive and infiltrated BMSC. Although several anticipated neuroregenerative, nerve guidance and angiogenic factor (e.g., bFGF, bone morphogenetic protein, angiopoietins, neural growth factor) genes exhibited an increased expression, a remarkable induction of genes for nerve guidance survival (e.g., cytokine receptor-like factor 1, glypican 1, Dickkopf homolog 2, osteopontin) was also noted. Conclusions- BMSC infiltrating the post-ischemic brain exhibit persistent epigenetic changes in gene expression for numerous extracellular genes, compared to their naive counterparts. These genes are relevant to the neuroprotection, regeneration ! and angiogenesis previously described following stem cell ther! apy in a nimal models of ischemic stroke. PMID: 20509949 [PubMed - as supplied by publisher] | | |
| Isolation and culture of adult mouse hepatocytes.
June 1, 2010 at 2:45 PM
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| | Isolation and culture of adult mouse hepatocytes. Methods Mol Biol. 2010;633:185-96 Authors: Li WC, Ralphs KL, Tosh D The liver performs a multitude of functions including the regulation of carbohydrate, fat, and protein metabolism, the detoxification of endo- and xenobiotics, and the synthesis and secretion of plasma proteins and bile. Isolated hepatocytes constitute a useful technique for studying liver function in an in vitro setting. Here we describe a method for the isolation of hepatocytes from adult mouse liver. The principle of the method is the two-step collagenase perfusion technique which involves sequential perfusion of the liver with ethylenediaminetetraacetic acid and collagenase. Following isolation, the cells can either be used for short-term studies or, alternatively, maintained in culture for prolonged periods to study long-term changes in gene expression. The protocol for mouse hepatocyte isolation may be applied to both normal and transgenic mice. PMID: 20204628 [PubMed - indexed for MEDLINE] | | |
| Isolation and culture of embryonic pancreas and liver.
June 1, 2010 at 2:45 PM
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| | Isolation and culture of embryonic pancreas and liver. Methods Mol Biol. 2010;633:91-9 Authors: Burke ZD, Li WC, Slack JM, Tosh D Culturing embryonic tissue in an in vitro setting offers the unique ability to manipulate the external medium and therefore to investigate the pathways involved in regulating normal organogenesis as well as providing models for developmental disorders. Here we describe a system for the in vitro culture of the dorsal pancreatic buds and liver buds from mouse embryos. The tissues are dissected from day 9.0 or 11.5 mouse embryos. The tissues are placed on fibronectin-coated coverslips in serum-containing medium and allowed to attach. Over the next few days, the buds grow as flattened structures which are thin enough to allow the use of wholemount immunostaining methods. PMID: 20204622 [PubMed - indexed for MEDLINE] | | |
| Isolation, culture, and characterisation of mouse embryonic oesophagus and intestine.
June 1, 2010 at 2:45 PM
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| | Isolation, culture, and characterisation of mouse embryonic oesophagus and intestine. Methods Mol Biol. 2010;633:81-90 Authors: Quinlan JM, Yu WY, Tosh D The gastrointestinal tract of vertebrates is lined by epithelium that develops from the endodermal germ layer. The oesophagus and intestine form part of the gastrointestinal tract and studying the normal development of both tissues is difficult due to lack of suitable in vitro model systems. One of the criteria for a reliable culture model includes the ability to carry out real-time observations in vitro. The method we describe here is based on the isolation of embryonic oesophagus and intestine from 11.5-day-old embryos and culture on fibronectin-coated coverslips in basal Eagle's medium and 20% fetal bovine serum. This model permits real-time observations of both tissues and over a few days in culture, markers of differentiation appear. This culture system appears to recapitulate normal oesophagus and intestine development. PMID: 20204621 [PubMed - indexed for MEDLINE] | | |
| An alternative technique to shape scaffolds with hierarchical porosity at physiological temperature.
June 1, 2010 at 2:45 PM
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| | An alternative technique to shape scaffolds with hierarchical porosity at physiological temperature. Acta Biomater. 2010 Apr;6(4):1288-96 Authors: Peña J, Román J, Victoria Cabañas M, Vallet-Regà M The method described in this work, termed GELPOR3D, is characterised by its simplicity of use, low-cost equipment, compositional flexibility, and lack of aggressive or toxic solvents or other thermal treatment. This technique ensures the generation of a three-dimensional network of interconnected pores (300-900 microm); in addition, a random and not necessarily connected porosity is generated, yielding a hierarchical porous architecture from the macro to the molecular scale. The interconnected pores, large enough to ensure an adequate vascularization and new tissue ingrowth, can be obtained by pouring a slurry containing a biodegradable thermogel (such as agarose and gellan) and a ceramic into a mold consisting of a three-dimensional network of rigid filaments. Additional pore distributions in the macropore region can be tailored as a function of the drying/preservation technology (10-100 microm) or the interaction between the inorganic particles coated by the pol! ymeric components (0.1-1 microm). Moreover, porosity in the mesopore range can be created by shaping ceramics such as mesoporous silica or nanocrystalline carbonatehydroxyapatite. In addition to the various bioceramics that have been successfully shaped, this method is flexible enough to allow the introduction of certain substances whose controlled release may help to avoid some negative effects that usually appear with the implantation of a material, i.e. infection, inflammation, etc., or to simplify some of the many steps required for the successful integration of a graft. PMID: 19887122 [PubMed - indexed for MEDLINE] | | |
| Synthesis and recovery characteristics of branched and grafted PNIPAAm-PEG hydrogels for the development of an injectable load-bearing nucleus pulposus replacement.
June 1, 2010 at 2:45 PM
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| | Synthesis and recovery characteristics of branched and grafted PNIPAAm-PEG hydrogels for the development of an injectable load-bearing nucleus pulposus replacement. Acta Biomater. 2010 Apr;6(4):1319-28 Authors: Thomas JD, Fussell G, Sarkar S, Lowman AM, Marcolongo M A family of injectable poly(N-isopropyl acrylamide) (PNIPAAm) copolymer hydrogels has been fabricated in order to tune mechanical properties to support load-bearing function and dimensional recovery for possible use as load-bearing medical devices, such as a nucleus pulposus replacement for the intervertebral disc. PNIPAAm-polyethylene glycol (PEG) copolymers were synthesized with varying hydrophilic PEG concentrations as grafted or branched structures to enhance dimensional recovery of the materials. Polymerizations were confirmed with attenuated total reflectance-Fourier transform infrared spectroscopy and proton nuclear magnetic resonance spectroscopy studies. Incorporation of PEG was effective in raising water content of pure PNIPAAm hydrogels (29.3% water for pure PNIPAAm vs. 47.7% for PEG branches and 39.5% for PEG grafts). PNIPAAm with 7% grafted as well as 7% branched PEG had significantly reduced compressive modulus compared to that of pure PNIPAAm. Initi! ally recovered compressive strain was significantly increased for 7% PEG branches after pre-testing immersion in PBS for up to 33 days, while 7% PEG grafts decreased this value. Sample height recovery for pure PNIPAAm was limited to 31.6%, while PNIPAAm with 7% branches was increased to 71.3%. When mechanically tested samples were allowed to recover without load over 30 min, each composition was able to significantly recover height, indicating that the time to recovery is slower than the unloading rates typically used in testing. While the incorporation of hydrophilic PEG was expected to alter the mechanical behavior of the hydrogels, only the branched form was able to significantly enhance dimensional recovery. PMID: 19837195 [PubMed - indexed for MEDLINE] | | | | 
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