Safety evaluation of Sapindus laurifolius leaf extract in Wistar rats

Objectives:The present work was aimed to study the phytochemical composition of the Sapindus laurifolius leaves andtoxicological effect of the Sapindus laurifolius leaf extract in a systematic way using Wistar albino rats as a model animal.Materials and Methods :The identification of phytoconstituents present in the leaf extract was performed using Highperformance thin layer chromatography (HPTLC). In toxicity studies, the acute oral toxicity study was conducted as per theguidelines of Organization for Economic Co-operation and Development (OECD 423 Acute Toxic Class Method) for testingof chemicals. In repeated dose 28-day oral toxicity study (OECD 407), methanolic leaf extract administered at the dose of 50,200 and 800 mg/kg BWand limit dose of 1000 mg/kg BW.Results: Saponins, flavanoids, glycosides and bitter principles were the major phytoconstituents identified. In acute toxicitystudy, the LD cut-off values were found to be more than 2g/kg in leaf extract. In repeated dose 28-day oral toxicity, significant 50(P<0.05) increase in AST, ALT, BUN and creatinine, significant (P<0.05) increase in total protein was noticed. Thehistopathological changes confined to liver, kidney and intestine, revealed mild to moderate hepatotoxicity, severenephrotoxicity and increased goblet cell activity. The changes were found to correlate with increased dose of leaf extract.Conclusion:The phytochemical analysis of Sapindus laurifolius revealed the presence of saponins, glycosides, flavonoidsand bitter principles.The acute oral toxicity study of S. laurifolius methanolic leaf extract in rats resulted in no toxicity even atthe highest dose, but in repeated 28-day oral toxicity study revealed mild to moderate hepatotoxicity, severe nephrotoxicityand intestinal damage

Effect of Sintering Temperature on Structural Properties of Cd doped Co-Zn Ferrite

This work has the objective of influence of sintering temperatures (800 °C and 1000 °C) structural properties of cadmium doped cobalt zinc ferrite (Co0.5Zn0.5Cd0.2Fe1.8O4) prepared by simple, low-cost Solid State reaction method. The structural parameters were explored by XRD technique. The X-ray analysis confirms the formation of particles of cubic inverse spinel structure. The structural and mechanical properties comparatively reported

Preparation, Characterization and In Vitro Drug Release Studies of 6-mercaptopurine Thin Film

Oral thin films of 6-mercaptopurine were fabricated from mucoadhesive polymer, chitosan and polyvinylpyrrolidone for the purpose of prolonging drug release and improving its bioavailability. All fabricated film formulations prepared were smooth and translucent, with good flexibility. The weight and thickness of all the formulations were found to be uniform. These films were also evaluated for surface pH, folding endurance, swelling percentage (% S) and in vitro disintegration time. Scanning Electron Microscopy (SEM) and Fourier Transform Infrared (FTIR) were used to evaluate the physico-chemical nature of the films. In-vitro drug release have shown enhanced release profiles for thin films compared to pure drug and the release patterns have been found to be pH dependant. The results of the study reveals that fabrication of 6-MP oral thin film by using solvent cast technology is a simple and an efficient method for drug delivery to achieve desired therapeutic compliance.Keywords: 6-mercaptopurine; In Vitro Drug Release; SEM; FTI

Studies on Certain Serum Metabolites in non Pregnant and Pregnant Bannur Ewes

Abstract A study was undertaken with the objective of making an insight in to the changes with respect to the level of certain metabolites in non pregnant and pregnant Bannur ewes which are indicative of their nutritional status and physiological well being. Eighteen Bannur ewes maintained under identical managemental conditions in a semi-intensive rearing farm which were of two to four years of age are categorized in to three groups, comprising of six animals in each group, such as non pregnant (Group I), early pregnant (Group II, at 20 to 35 days of pregnancy) and late pregnant (Group III, at 105 to 120 days of pregnancy) ewes based upon ultrasound imaging technique. The metabolites such as serum levels of β-hydroxybutyrate, glucose, total protein, blood urea nitrogen and lipid profile components were determined at 0, 7 and 15 th day of sample collection. The biochemical parameters did not vary significantly (P&gt;0.05) between the groups except for the glucose levels which were significantly (P&lt;0.05) lower in Group III compared to Group I and Group II. It was concluded that, all the parameters studied in Bannur breed of ewes during non pregnancy, early pregnancy and late pregnancy were within the normal reference range described for sheep and it was suggestive that none of the ewes in the present study suffered from any kind of metabolic disorders

Effect of food matrix and processing on release of almond protein during simulated digestion

Abstract The aims of the present work were to assess digestibility of almond protein in the upper gastrointestinal tract, evaluate the effects of food matrix on protein release and assess the persistence of immunoreactive polypeptides generated during simulated digestion. Prunin, the most abundant protein in almond flour, was sensitive to pepsin, with complete digestion after 20 min in the gastric phase. Addition of the surfactant phosphatidylcholine did not affect the rate and kinetic of digestion, as observed by SDS-PAGE analysis and HPLC, in the stomach and the small intestine of either natural or blanched almond flour. However, incorporation of almond flour into a food matrix, such as chocolate mousse and Victorian sponge cake, decreased the rate of almond protein degradation by pepsin and immunoreactivity of almond polypeptides detected by dot blots and sandwich ELISA retained better. Most of the almond protein identified by in-gel tryptic digestion and MALDI-TOF analysis corresponded to prunin, with pI values of 5–7. Further human sera studies are warranted to investigate the relationship between food matrix and almond allergy

Silencing hepatic MCJ attenuates non-alcoholic fatty liver disease (NAFLD) by increasing mitochondrial fatty acid oxidation

Nonalcoholic fatty liver disease (NAFLD) is considered the next major health epidemic with an estimated 25% worldwide prevalence. No drugs have yet been approved and NAFLD remains a major unmet need. Here, we identify MCJ (Methylation-Controlled J protein) as a target for non-alcoholic steatohepatitis (NASH), an advanced phase of NAFLD. MCJ is an endogenous negative regulator of the respiratory chain Complex I that acts to restrain mitochondrial respiration. We show that therapeutic targeting of MCJ in the liver with nanoparticle- and GalNAc-formulated siRNA efficiently reduces liver lipid accumulation and fibrosis in multiple NASH mouse models. Decreasing MCJ expression enhances the capacity of hepatocytes to mediate beta -oxidation of fatty acids and minimizes lipid accumulation, which results in reduced hepatocyte damage and fibrosis. Moreover, MCJ levels in the liver of NAFLD patients are elevated relative to healthy subjects. Thus, inhibition of MCJ emerges as an alternative approach to treat NAFLD. Non-alcoholic fatty liver (NAFLD) disease causes degeneration of the liver, affects about 25% of people globally, and has no approved treatment. Here, the authors show that the therapeutic siRNA-driven silencing of MCJ in the liver is an effective and safe treatment for NAFLD in multiple mouse models.We thank Douglas Taatjes and Nicole Bouffard for help with confocal microscopy analysis (Microscopy Imaging Center) at the University of Vermont. We also thank the University of Vermont Medical Center's Department of Pathology and Laboratory Medicine Histology and Clinical Laboratories for assistance with liver section staining and AST/ALT measurement, respectively. This work was supported by NIH STTR R41DK112429 (M.R.), NIH PO GM103496 (M.R.), Mitotherapeutix LLC (M.R., K.F, and M.L.M.-C.), MINECO/Feder SAF2015-65327-R and RTI2018-096494-B-100 (J.A.), MINECO/Feder SAF2017-87301-R (M.L.M-C.), BIOEF (M.L.M.-C.), EITB Maratoia BIO15/CA/014 (M.L.M-C), BBVA (M.L.M.-C.), La Caixa Foundation (M.L.M.-C.), Basque Country Health Department 2013111114 (M.L.M-C), MINECO/Feder SAF2015-64352-R (P.A.) and MINECO-Feder RTI2018-095134-B-100 (P.A.). ISCIII-Feder PI17/00535 (C.G.-M.), ISCIII-Feder CP14/00181, and PI16/00823 (A.G-R.), and Francisco Cobos Foundation (A.G.-R.). CIC bioGUNE is the recipient of a Severo Ochoa Excellence Accreditation (SEV-2016-0644) by the Ministry of Science, Innovation and Universities

Cystatin E/M suppresses legumain activity and invasion of human melanoma

<p>Abstract</p> <p>Background</p> <p>High activity of cysteine proteases such as legumain and the cathepsins have been shown to facilitate growth and invasion of a variety of tumor types. In breast cancer, several recent studies have indicated that loss of the cysteine protease inhibitor cystatin E/M leads to increased growth and metastasis. Although cystatin E/M is normally expressed in the skin, its role in cysteine protease regulation and progression of malignant melanoma has not been studied.</p> <p>Methods</p> <p>A panel of various non-melanoma and melanoma cell lines was used. Cystatin E/M and C were analyzed in cell media by immunoblotting and ELISA. Legumain, cathepsin B and L were analyzed in cell lysates by immunoblotting and their enzymatic activities were analyzed by peptide substrates. Two melanoma cell lines lacking detectable secretion of cystatin E/M were transfected with a cystatin E/M expression plasmid (pCST6), and migration and invasiveness were studied by a Matrigel invasion assay.</p> <p>Results</p> <p>Cystatin E/M was undetectable in media from all established melanoma cell lines examined, whereas strong immunobands were detected in two of five primary melanoma lines and in two of six lines derived from patients with metastatic disease. Among the four melanoma lines secreting cystatin E/M, the glycosylated form (17 kD) was predominant compared to the non-glycosylated form (14 kD). Legumain, cathepsin B and L were expressed and active in most of the cell lines, although at low levels in the melanomas expressing cystatin E/M. In the melanoma lines where cystatin E/M was secreted, cystatin C was generally absent or expressed at a very low level. When melanoma cells lacking secretion of cystatin E/M were transfected with pCST6, their intracellular legumain activity was significantly inhibited. In contrast, cathepsin B activity was not affected. Furthermore, invasion was suppressed in cystatin E/M over-expressing melanoma cell lines as measured by the transwell Matrigel assay.</p> <p>Conclusions</p> <p>These results suggest that the level of cystatin E/M regulates legumain activity and hence the invasive potential of human melanoma cells.</p

BubR1 as a prognostic marker for recurrence-free survival rates in epithelial ovarian cancers

BACKGROUND: Epithelial ovarian cancer is one of the most lethal malignancies, and has a high recurrence rate. Thus, prognostic markers for recurrence are crucial for the care of ovarian cancer. As ovarian cancers frequently exhibit chromosome instability, we aimed at assessing the prognostic significance of two key mitotic kinases, BubR1 and Aurora A. METHODS: We analysed paraffin-embedded tissue sections from 160 ovarian cancer patients whose clinical outcomes had been tracked after first-line treatment. RESULTS: The median recurrence-free survival in patients with a positive and negative expression of BubR1 was 27 and 83 months, respectively (Po0.001). A positive BubR1 expression was also associated with advanced stage, serous histology and high grade. In contrast, Aurora A immunostaining did not correlate with any of the clinical parameters analysed. CONCLUSION: BubR1, but not Aurora A, is a prognostic marker for recurrence-free survival rates in epithelial ovarian cancers.Research in the H Lee laboratory is funded by the National Research Laboratory Program from the Korean ministry of Education and Science (ROA-2008-000-20023-0). This work was also supported by the Seoul National University Hospital Grant (0420080450), the 21C Frontier Functional Genome Project (FG06- 2-14) of the Korean ministry of Education and Science, Korea Research Foundation (KRF-2005-C00097), and the National R&D Program for Cancer Control (0620070) from the Korean ministry of Health welfare and Family Affairs. Imaging facilities in the H Lee laboratory are funded by RCFC (R11-2005-009-04003-0) of the SRC program from KOSEF