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Liver is a vital organ and is routinely exposed to toxins. Carbon tetrachloride is one such noxious agent which cause toxicity in liver when CYP450 enzyme bio-activates it. Many hepatoprotective agents are available in market with severe side effects. Appropriate agent is required to combat such liver problems. Azole compounds have much therapeutic values in many diseases. Based upon this fact, present study is aimed to evaluate the repurposing of Itraconazole in the prevention of hepatic fibrosis via inhibition of cytochrome P450 pathway. For in-vitro evaluation of cyto-protective effects in HepG2 cells (untreated and treated groups), cell viability assays, antioxidant evaluation, enzyme linked immunosorbent assay (ELISA) and immunocytochemistry was used. For in-vivo evaluation, CCl4 induced liver fibrotic rat model was used and post treated evaluation was done by blood biochemistry, hematoxylin and eosin (H&E) staining and gene expression profiling. Results of the current study indicated hepatoprotective role of itraconazole via inhibition of CYP450 pathway inhibition. Therefore, Itraconazole use could be a potential therapeutic approach to prevent liver fibrosis. [ABSTRACT FROM AUTHOR]

Cellular therapies hold promise to alleviate liver diseases. This study explored the potential of allogenic serum isolated from rat with acute CCl4injury to differentiate adipose derived stem cells (ADSCs) towards hepatic lineage. Acute liver injury was induced by CCl4which caused significant increase in serum levels of VEGF, SDF1α and EGF. ADSCs were preconditioned with 3% serum isolated from normal and acute liver injury models. ADSCs showed enhanced expression of hepatic markers (AFP, albumin, CK8 and CK19). These differentiated ADSCs were transplanted intra-hepatically in CCl4-induced liver fibrosis model. After one month of transplantation, fibrosis and liver functions (alkaline phosphatase, ALAT and bilirubin) showed marked improvement in acute injury group. Elevated expression of hepatic (AFP, albumin, CK 18 and HNF4a) and pro survival markers (PCNA and VEGF) and improvement in liver architecture as deduced from results of alpha smooth muscle actin, Sirius red and Masson’s trichome staining was observed. [ABSTRACT FROM PUBLISHER]

Detailed knowledge of the three-dimensional (3D) structure of a protein is essential for the proper understanding of its function(s) that could be modified through posttranslational modifications (PTMs). Among these PTMs, alterations of serine/threonine residues of a protein through phosphorylation and O-glycosylation are extremely dynamic and could modulate the functions of a protein by affecting their 3D structure. Potential of a protein for certain PTMs could be evaluated through computer-based methods. Erythropoietin (EPO) is a multifunctional protein that primarily regulates red blood cell production and is also involved in other nonhematopoietic functions; for example, EPO also has cardioprotective and neuroprotective effects. In this study, multifunctional EPO behavior has been revealed based on transient modifications of its receptor. In this study, PTMs of erythropoietin receptor (EPO-R) were predicted using neural network tools, and the possible effects of these modifications are suggested. Phosphorylation and O-glycosylation at serine 380 and 444 of the cytoplasmic domain of EPO-R seem to have an antagonistic role in controlling signaling events induced by EPO. O-glycosylation at threonine 423 might hinder β-TrCP (a ubiquitin ligase) binding, which ubiquitinates at K 428, and ultimately results in the recycling of EPO-R, thus increasing EPO sensitivity. In contrast, the phosphorylated form of the same residue inhibits the recycling of EPO-R and thereby decreases the EPO sensitivity. Additionally, the interplay of O-glycosylation modification at serine 478 and phosphorylation at tyrosine 479 might help in controlling the duration of EPO-induced signaling. [ABSTRACT FROM AUTHOR]

Hepatic oval cells are likely to be activated during advanced stage of liver fibrosis to reconstruct damaged hepatic tissue. However, their scarcity, difficulties in isolation, and in vitro expansion hampered their transplantation in fibrotic liver. This study was aimed to investigate the repair potential of in vitro differentiated hepatic oval-like cells in CCl4-induced liver fibrosis. BMSCs and oval cells were isolated and characterized from C57BL/6 GFP+ mice. BMSCs were differentiated into oval cells by preconditioning with HGF, EGF, SCF, and LIF and analyzed for the oval cells-specific genes. Efficiency of oval cells to reduce hepatocyte injury was studied by determining cell viability, release of LDH, and biochemical tests in a co-culture system. Further, in vivo repair potential of differentiated oval cells was determined in CCl4-induced fibrotic model by gene expression analysis, biochemical tests, mason trichrome, and Sirius red staining. Differentiated oval cells expressed hepatic oval cells-specific markers AFP, ALB, CK8, CK18, CK19. These differentiated cells when co-cultured with injured hepatocytes showed significant hepato-protection as measured by reduction in apoptosis, LDH release, and improvement in liver functions. Transplantation of differentiated oval cells like cells in fibrotic livers exhibited enhanced homing, reduced liver fibrosis, and improved liver functions by augmenting hepatic microenvironment by improved liver functions. This preconditioning strategy to differentiate BMSCs into oval cell leads to improved survival and homing of transplanted cells. In addition, reduction in fibrosis and functional improvement in mice with CCl4-induced liver fibrosis was achieved. [ABSTRACT FROM AUTHOR]

Tendinopathy or tendon degeneration in equine is an illness caused by inflammation or injury. Being a difficult tissue to repair, tendon damage can cause physical disabilities. Successful tendon regeneration requires an appropriate in-vitro model for tissue injury. Current study was planned to compare the effect of two inflammatory mediators: prostaglandinE2 (PGE2) and hydrogen peroxide (H2O2) on equine tendon fibroblasts. Tendon fibroblast cultures were subjected to two different doses of PGE2 (100ng/ml and 200ng/ml) and H2O2 (500µM and 1mM). Compared with untreated tendon fibroblasts, all doses of PGE2 and H2O2 reduced cell viability markedly. Viability was assessed by trypan blue staining method and crystal violet staining. Lactate dehydrogenase (LDH) release was measured to assess injury and cell proliferation by MTT assay. In comparison of dose, 100ng/ml of PGE2 and 500µM of H2O2 showed optimal results, whereas, higher dose exhibited highly reduced cell viability and increased cytotoxicity, which may cause difficulty in studying tendon injury in-vitro model. These findings suggest that both PGE2 and H2O2 have equal potential to cause tendon fibroblasts injury. Therefore, H2O2 can be used as cost-effective in-vitro model for tendon degeneration studies. [ABSTRACT FROM AUTHOR]

Background: To determine the antimicrobial, antiproliferative and antioxidant activities of ethanolic, ethyl acetate and petroleum ether extracts of Psidiumguajava leaves. Methods: The antimicrobial activity was assessed using disc diffusion method on Staphylococcus aureus, Bacillus cereus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli and Salmonella typhi. The anti-proliferative activity was determined on HeLa cell lines. The anti-proliferative activity was assessed using MTT assay while antioxidant activity was determined using catalase and superoxide dismutase (SOD) assays. Results: The MTT results indicate that ethyl acetate and petroleum ether extracts have significantly inhibited the proliferation of HeLa cells as compared to the control. Antioxidant assays showed that catalase activity was insignificant for all extracts however the SOD activity was significant. All the extracts showed inhibitory zones against B. cereus while only ethanolic extract of P. guajava leaves was active against S. aureus. Conclusion: Psidiumguajava plant leaves extracts can be used as antiproliferative agent against HeLa cell line. [ABSTRACT FROM AUTHOR]