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Vivo Results (vivo + result)
Selected AbstractsDeletion of the LIME adaptor protein minimally affects T and B cell development and functionEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 11 2007Claude Grégoire Abstract LIME (Lck-interacting membrane protein) is a transmembrane adaptor that associates with the Lck and Fyn protein tyrosine kinases and with the C-terminal Src kinase (Csk). To delineate the role of LIME in vivo, LIME-deficient mice were generated. Although Lime transcripts were expressed in immature and mature B and T cells, the absence of LIME impeded neither the development nor the function of B and T cells. TCR transgenic mice deprived of LIME showed, however, a 1.8-fold enhancement in positive selection. Since B cells and activated T cells express LIME and the related adaptor NTAL, mice lacking both adaptors were generated. Double-deficient mice showed no defect in the development and function of B and T cells, and the lack of LIME had no effect on the autoimmune syndrome that develops in aged NTAL-deficient mice. In contrast to a previous report, we further showed that this autoimmune syndrome develops in the absence of T cells. Therefore, our in vivo results refute all the previous roles postulated for LIME on the basis of studies of transformed B and T cells and demonstrate that LIME has no seminal role in the signaling cassette operated by antigen receptors and coreceptors. [source] Potentiation of isoniazid-induced liver toxicity by rifampicin in a combinational therapy of antitubercular drugs (rifampicin, isoniazid and pyrazinamide) in Wistar rats: A toxicity profile studyHEPATOLOGY RESEARCH, Issue 10 2007Sheikh Abdullah Tasduq Aim:, Biochemical characterization of long-term toxic manifestations of anti-tubercular (anti-TB) drugs , rifampicin (RIF), isoniazid (INH) and pyrazinamide (PZA) , individually and in two combinations: (i) RIF + INH, and (ii) RIF + INH + PZA in Wistar rats. Methods:, Animals received anti-TB drugs , alone or in combination , once daily p.o. for up to 90 days (doses, in mg/kg: RIF, 250; INH, 50; PZA, 100). Assays for alanine aminotransferase (ALT), alkaline phosphatase (ALP), bilirubin (serum) and lipid peroxidation (LPO), glutathione (GSH), glutathione peroxidase (GPx), catalase, Na+K+-ATPase and CYP 2E1 (liver) were performed to assess liver toxicity. Clinical biochemistry was done by commercial kits. Determinations were made at 0, 15, 30 and 90 days of treatment schedule. Results:, Anti-TB drugs-treated animals showed abnormal rises or falls (>1.5,2 fold) in the serum/liver parameters. Mild hyperlipidemia, hypercholesterolemia and hyperuricemia were the other pathologies. Of all the treated groups, INHalone or in combination with other drugs produced a progressive enhancement of toxicity over 15,90 days. The in vivo results were further supported by in vitro results (MTT assay, GSH and LPO) in primary cultures of rat hepatocyte. Results indicated that anti-TB drugs in combination: (i) caused membrane damage resulting in leakage of ALT, ALP and bilirubin; (ii) caused imbalance in endogenous enzymatic oxidant,antioxidant defense via increased lipid peroxidation and in glutathione homeostasis; and (iii) enhanced the CYP 2E1-mediated bioactivation mechanism. Conclusion:, Toxicity manifestations seemed to be heptocytic injury targeted at hepatocytes, bile ducts or sinusoidal cells related to hepatitis and primary biliary cholestasis. [source] Induction of hepatic differentiation of mouse bone marrow stromal stem cells by the histone deacetylase inhibitor VPAJOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 8b 2009Ye Chen Abstract Bone marrow stromal stem cells (BMSSCs) may have potential to differentiate in vitro and in vivo into hepatocytes. Here, we investigated the effects of valproic acid (VPA) involved in epigenetic modification, a direct inhibitor of histone deacetylase, on hepatic differentiation of mouse BMSSCs. Following the treatment of 2.5 mM VPA for 72 hrs, the in vitro expanded, highly purified and functionally active mouse BMSSCs from bone marrow were either exposed to some well-defined cytokines and growth factors in a sequential way (fibroblast growth factor-4 [FGF-4], followed by HGF, and HGF + OSM + ITS + dexamethasone, resembling the order of secretion during liver embryogenesis) or transplanted (caudal vein) in mice submitted to a protocol of chronic injury (chronic i.p. injection of CCl4). Additional exposure of the cells to VPA considerably improved the in vitro differentiation, as demonstrated by a more homogeneous cell population exhibited epithelial morphology, increasing expression of hepatic special genes and enhanced hepatic functions. Further more, in vivo results indicate that the pre-treatment of VPA significantly increased the homing efficiency of BMSSCs to the site of liver injury and, additionally, for supporting hepatic differentiation as well as in vitro. We have demonstrated the usefulness of VPA in the transdifferentiation of BMSSCs into hepatocytes both in vitro and in vivo, and regulation of fibroblast growth factor receptors (FGFRs) and c-Met gene expression through post-translational modification of core histones might be the primary initiating event for these effects. This mode could be helpful for liver engineering and clinical therapy. [source] Enhanced type I interferon signaling and recruitment of chemokine receptor CXCR3-expressing lymphocytes into the skin following treatment with the TLR7-agonist imiquimodJOURNAL OF CUTANEOUS PATHOLOGY, Issue 4 2005Joerg Wenzel Introduction:, Imiquimod (AldaraÔ) is an immune response modifier approved for the topical treatment of external genital and perianal warts which can mediate regression of several cutaneous malignancies [basal cell carcinoma (BCC), Bowen's disease, actinic keratosis, and metastasis of malignant melanoma]. Recently, it was discovered that imiquimod acts through the toll-like receptor (TLR) 7. We hypothesize that TLR7-signaling strongly induces the production of interferon (IFN) ,, which is able to enhance Th1-mediated cellular antiviral and antitumor immunity. Patients and methods:, In the present study we analyzed the expression of MxA, a protein specifically induced by type I IFNs during topical imiquimod treatment in several patients suffering from different cutaneous malignancies (BCC, cutaneous metastasis of melanoma, and breast cancer), and characterized the inflammatory infiltrate, along with the expression of chemokine receptor CXCR3, by immunohistochemistry. Results:, Treatment with the TLR7-agonist imiquimod induced a significant lesional lymphocytic inflammation, associated with strong expression of MxA, indicating the induction of type I IFN signaling. The extent of lesional MxA staining closely correlated with the number of infiltrating T lymphocytes and the expression of the chemokine receptor CXCR3, characteristic for Th1-biased immune responses. Discussion:, Our in vivo results suggest an important role for TLR7-induced production of type I IFN, which links innate and adaptive immunity and promotes specific Th1-biased cellular immune response capable of eliminating cutaneous malignancies. MxA appears to be a valuable parameter to demonstrate IFN-type I expression in imiquimod therapy. [source] Molecular architecture of myelinated peripheral nerves is supported by calorie restriction with agingAGING CELL, Issue 2 2009Sunitha Rangaraju Summary Peripheral nerves from aged animals exhibit features of degeneration, including marked fiber loss, morphological irregularities in myelinated axons and notable reduction in the expression of myelin proteins. To investigate how protein homeostatic mechanisms change with age within the peripheral nervous system, we isolated Schwann cells from the sciatic nerves of young and old rats. The responsiveness of cells from aged nerves to stress stimuli is weakened, which in part may account for the observed age-associated alterations in glial and axonal proteins in vivo. Although calorie restriction is known to slow the aging process in the central nervous system, its influence on peripheral nerves has not been investigated in detail. To determine if dietary restriction is beneficial for peripheral nerve health and glial function, we studied sciatic nerves from rats of four distinct ages (8, 18, 29 and 38 months) kept on an ad libitum (AL) or a 40% calorie restricted diet. Age-associated reduction in the expression of the major myelin proteins and widening of the nodes of Ranvier are attenuated by the dietary intervention, which is paralleled with the maintenance of a differentiated Schwann cell phenotype. The improvements in nerve architecture with diet restriction, in part, are underlined by sustained expression of protein chaperones and markers of the autophagy,lysosomal pathway. Together, the in vitro and in vivo results suggest that there might be an age-limit by which dietary intervention needs to be initiated to elicit a beneficial response on peripheral nerve health. [source] Multiecho reconstruction for simultaneous water-fat decomposition and T2* estimation,JOURNAL OF MAGNETIC RESONANCE IMAGING, Issue 4 2007Huanzhou Yu PhD Abstract Purpose To describe and demonstrate the feasibility of a novel multiecho reconstruction technique that achieves simultaneous water-fat decomposition and T2* estimation. The method removes interference of water-fat separation with iron-induced T2* effects and therefore has potential for the simultaneous characterization of hepatic steatosis (fatty infiltration) and iron overload. Materials and Methods The algorithm called "T2*-IDEAL" is based on the IDEAL water-fat decomposition method. A novel "complex field map" construct is used to estimate both R2* (1/T2*) and local B0 field inhomogeneities using an iterative least-squares estimation method. Water and fat are then decomposed from source images that are corrected for both T2* and B0 field inhomogeneity. Results It was found that a six-echo multiecho acquisition using the shortest possible echo times achieves an excellent balance of short scan and reliable R2* measurement. Phantom experiments demonstrate the feasibility with high accuracy in R2* measurement. Promising preliminary in vivo results are also shown. Conclusion The T2*-IDEAL technique has potential applications in imaging of diffuse liver disease for evaluation of both hepatic steatosis and iron overload in a single breath-hold. J. Magn. Reson. Imaging 2007;26:1153,1161. © 2007 Wiley-Liss, Inc. [source] Calculation of cerebral perfusion parameters using regional arterial input functions identified by factor analysisJOURNAL OF MAGNETIC RESONANCE IMAGING, Issue 4 2006Linda Knutsson MS Abstract Purpose To calculate regional cerebral blood volume (rCBV), regional cerebral blood flow (rCBF), and regional mean transit time (rMTT) accurately, an arterial input function (AIF) is required. In this study we identified a number of AIFs using factor analysis of dynamic studies (FADS), and performed the cerebral perfusion calculation pixel by pixel using the AIF that was located geometrically closest to a certain voxel. Materials and Methods To verify the robustness of the method, simulated images were generated in which dispersion or delay was added in some arteries and in the corresponding cerebral gray matter (GM), white matter (WM), and ischemic tissue. Thereafter, AIFs were determined using the FADS method and simulations were performed using different signal-to-noise ratios (SNRs). Simulations were also carried out using an AIF from a single pixel that was manually selected. In vivo results were obtained from normal volunteers and patients. Results The FADS method reduced the underestimation of rCBF due to dispersion or delay that often occurs when only one AIF represents the entire brain. Conclusion This study indicates that the use of FADS and the nearest-AIF method is preferable to manual selection of one single AIF. J. Magn. Reson. Imaging 2006. © 2006 Wiley-Liss, Inc. [source] Magnetic resonance brain perfusion imaging with voxel-specific arterial input functionsJOURNAL OF MAGNETIC RESONANCE IMAGING, Issue 3 2006Renate Grüner MSc Abstract Purpose To propose an automatic method for estimating voxel-specific arterial input functions (AIFs) in dynamic contrast brain perfusion imaging. Materials and Methods Voxel-specific AIFs were estimated blindly using the theory of homomorphic transformations and complex cepstrum analysis. Wiener filtering was used in the subsequent deconvolution. The method was verified using simulated data and evaluated in 10 healthy adults. Results Computer simulations accurately estimated differently shaped, normalized AIFs. Simple Wiener filtering resulted in underestimation of flow values. Preliminary in vivo results showed comparable cerebral flow value ratios between gray matter (GM) and white matter (WM) when using blindly estimated voxel-specific AIFs or a single manually selected AIF. Significant differences (P , 0.0125) in mean transit time (MTT) and time-to-peak (TTP) in GM compared to WM was seen with the new method. Conclusion Initial results suggest that the proposed method can replace the tedious and difficult task of manually selecting an AIF, while simultaneously providing better differentiation between time-dependent hemodynamic parameters. J. Magn. Reson. Imaging 2006. © 2006 Wiley-Liss, Inc. [source] A composite material model for improved bone formationJOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, Issue 7 2010Silvia Scaglione Abstract The combination of synthetic polymers and calcium phosphates represent an improvement in the development of scaffolds for bone-tissue regeneration. Ideally, these composites provide both mechanically and architecturally enhanced performances; however, they often lack properties such as osteoconductivity and cell bioactivation. In this study we attempted to generate a composite bone substitute maximizing the available osteoconductive surface for cell adhesion and activity. Highly porous scaffolds were prepared through a particulate leaching method, combining poly-,-caprolactone (PCL) and hydroxyapatite (HA) particles, previously coated with a sucrose layer, to minimize their embedding by the polymer solution. Composite performances were evaluated both in vitro and in vivo. In PCL,sucrose-coated HA samples, the HA particles were almost completely exposed and physically distinct from the polymer mesh, while uncoated control samples showed ceramic granules massively covered by the polymer. In vivo results revealed a significant extent of bone deposition around all sucrose-coated HA granules, while only parts of the control uncoated HA granules were surrounded by bone matrix. These findings highlight the possibility of generating enhanced osteoconductive materials, basing the scaffold design on physiological and cellular concepts. Copyright © 2010 John Wiley & Sons, Ltd. [source] Regulation of prostaglandin synthesis in ovaries of sexually-mature zebrafish (Danio rerio)MOLECULAR REPRODUCTION & DEVELOPMENT, Issue 11 2009Andrea L. Lister This study investigates the regulation of prostaglandin (PG) synthesis in the ovaries of sexually-mature zebrafish (Danio rerio). We examined the ovarian expression of genes within the arachidonic acid (AA) pathway, and the ovarian levels of 17,,20,-dihydroxy-4-pregnen-3-one (17,,20,-P), 17,-estradiol (E2), and PGF2, in spawning and nonspawning fish during the ovulatory cycle. Real-time RT-PCR analysis revealed that the expression levels of cytosolic phospholipase A2 (cpla2) and cyclooxygenases (COX)-2 (ptgs2) in ovarian fragments and in isolated full-grown follicles of spawning fish were highest at 6:00 when ovulation was expected to occur. In nonspawning fish, cpla2 expression levels declined over time while ptgs2 expression displayed the same temporal pattern as in spawning fish. Elevated levels of 17,,20,-P in the spawning fish occurred at 3:30, but there were no changes in the nonspawning fish. In other studies conducted to investigate the hormonal regulation of AA pathway genes, fish exposed via the water for 24 or 96,hr to 17,,20,-P or E2 exhibited reduced ovarian expression levels of COX-1 (ptgs1) and PG E synthase-2 (ptgsl), and E2 reduced the expression of cpla2. Injection of human chorionic gonadotropin (hCG) (100,IU) led to increased expression levels of cpla2 and ptgs2 at 2 and 18,hr post-treatment, but consistently reduced ptgs1 and ptgsl expression. In these fish, ovarian levels of 17,,20,-P were elevated at all time points and PGF2, levels in the hCG-treated group were significantly higher than the control fish at 18,hr. Collectively, these in vivo results suggest that gonadotropins and steroids are involved in the regulation of the AA pathway in ovarian follicles of zebrafish. Mol. Reprod. Dev. 76: 1064,1075, 2009. © 2009 Wiley-Liss, Inc. [source] Brain GABA editing by localized in vivo1H magnetic resonance spectroscopyNMR IN BIOMEDICINE, Issue 2 2004G. Bielicki Abstract Editing of GABA by 1H MRS in a specific brain area is a unique tool for in vivo non-invasive investigation of neurotransmission disorders. Selective GABA detection is achieved using sequences based on double quantum coherence (DQC). Our pulse sequence makes accurate measurements without artefacts due to spatial localization. The sequence was tested on a phantom solution. The effect of vigabatrin, a specific inhibitor of GABA transaminase, was measured in rat brain and GABA detection was performed in vivo in monkey brain using this procedure. Rats were spilt into two groups. In the control group, the rats had access to water and, in the other group (vigabatrin, VGB, rats), animals were allowed free access to drinking water containing vigabatrin. After 3 weeks of treatment, rats were anesthetized for in vivo NMR spectroscopy investigation. At the end of the experiment, brains were quickly removed, freeze-clamped and extracted with 4% perchloric acid. One part of the acid extract was used for GABA concentrations assessment by ion exchange chromatography with ninhydrin detection. The second was used for high-resolution NMR analysis. By chromatography measurements, the GABA concentration was 1.23±0.06,,mol/g for controls, while for vigabatrin-treated rats the GABA concentration was 4.89±1.60,,mol/g. The NMR in vivo results were closely correlated with the NMR ex vivo (r=0.99, p<0.01) and chromatography results (r=0.98, p<0.01). The correlation between ex vivo results and chromatography results was also high (r=0.99, p<0.001). This pulse sequence performed GABA editing from a 376,,l voxel located on the right basal ganglia area in a non-human primate brain. This in vivo GABA editing scheme can thus be proposed for accurate measurement of brain GABA concentrations. Copyright © 2004 John Wiley & Sons, Ltd. [source] Functional and Biocompatibility Performances of an Integrated Maglev Pump-OxygenatorARTIFICIAL ORGANS, Issue 1 2009Tao Zhang Abstract To provide respiratory support for patients with lung failure, a novel compact integrated pump-oxygenator is being developed. The functional and biocompatibility performances of this device are presented. The pump-oxygenator is designed by combining a magnetically levitated pump/rotor with a uniquely configured hollow fiber membrane bundle to create an assembly free, ultracompact, all-in-one system. The hemodynamics, gas transfer and biocompatibility performances of this novel device were investigated both in vitro in a circulatory flow loop and in vivo in an ovine animal model. The in vitro results showed that the device was able to pump blood flow from 2 to 8 L/min against a wide range of pressures and to deliver an oxygen transfer rate more than 300 mL/min at a blood flow of 6 L/min. Blood damage tests demonstrated low hemolysis (normalized index of hemolysis [NIH],0.04) at a flow rate of 5 L/min against a 100-mm Hg afterload. The data from five animal experiments (4 h to 7 days) demonstrated that the device could bring the venous blood to near fully oxygen-saturated condition (98.6% ± 1.3%). The highest oxygen transfer rate reached 386 mL/min. The gas transfer performance was stable over the study duration for three 7-day animals. There was no indication of blood damage. The plasma free hemoglobin and platelet count were within the normal ranges. No gross thrombus is found on the explanted pump components and fiber surfaces. Both in vitro and in vivo results demonstrated that the newly developed pump-oxygenator can achieve sufficient blood flow and oxygen transfer with excellent biocompatibility. [source] | |||||||||||||