Standardization of a traditional polyherbo-mineral formulation Brahmi vati

The present study deals with standardization of an in-house standard preparation and three marketed samples of Brahmi vati, which is a traditional medicine known to be effective in mental disorders, convulsions, weak memory, high fever and hysteria. Preparation and standardization have been done by following modern scientific quality control proceduresfor raw material and the finished products. The scanning electron microscopic (SEM) analysis showed the reduction of metals and minerals (particle size range 2-5 ìm) which indicates the proper preparation of bhasmas, the important ingredient of Brahmi vati. Findings of EDX analysis of all samples of Brahmi vati suggested the absence of Gold, an importantconstituent of Brahmi vati in two marketed samples. All the samples of Brahmi vati were subjected to quantitative estimation of Bacoside A (marker compound) by HPTLC technique. Extraction of the samples was done in methanol and the chromatograms were developed in Butanol: Glacial acetic acid: water (4.5:0.5:5 v/v) and detected at 225nm. The regression analysis of calibration plots of Bacoside A exhibited linear relationship in the concentration range of 50-300 ng, while the %recovery was found to be 96.06% w/w, thus proving the accuracy and precision of the analysis. The Bacoside A content in the in-house preparation was found to be higher than that of the commercial samples. The proposed HPTLC method was found to be rapid, simple and accurate for quantitative estimation of Bacoside A in different formulations. The results of this study could be used as a model data in the standardization of Brahmi vati.Keywords: Brahmi vati( BV); Standardization; EDX; HPTLC; Bacoside-

Fermentable sugars and microbial inhibitors formation from two-stage pretreatment of corn stalk with variation in particle size and severity factor

Microbial inhibitors including weak acids, furan derivatives and phenolic compounds are key problems of cellulosic bio-fuels production by fermentation. Most of these inhibitors are sugars and lignin degradation compounds, which are almost unavoidable during pretreatment processes. While, most of the one stage pretreatment has been conducted at high severity factors of 3.5 or more to get high sugar yield, with increase in severity factor, high concentration of microbial inhibitors were formed and significantly affected downstream biofuel yield. Thus, a two-stage pretreatment of corn stalk, hydrothermal followed by oxalic acid, under low severity factor and its enzymatic degradability was investigated in this study to identify fermentable sugar production and corresponding microbial inhibitors formation. Additionally, effect of equivalent severity factors of 2 to 3.5 and particle sizes of 1 to 35 mm were also studied systematically. Particle size of 15 mm was found as an optimum size at an equivalent severity factor of 2.5. Sugars 61.99 ± 0.03 g and inhibitors 5.12 ± 0.01 g from 100 g of corn stalk were obtained at the optimum particle size and pretreatment condition. The highest glucan conversion and recovery at the optimum conditions were 92.95± 0.08 and 78.42± 0.07%, respectively. Overall, the two-stage pretreatment process with the larger particle size and low equivalent severity factor could be an alternative to reduce microbial inhibitors formation and excessive biomass processing cost.Key words: Bio-fuel, corn stalk, pretreatment, particle size, microbial inhibitors, fermentable sugars

Evaluation of Antitumor Activity of Cuscuta Reflexa Roxb (Cuscutaceae) Against Ehrlich Ascites Carcinoma in Swiss Albino Mice

Purpose: The aim of this study was to investigate the antitumor effect of the chloroform and ethanol extract of the whole plant of Cuscuta reflexa Roxb. (Cuscutaceae) in Swiss albino mice against Ehrlich Ascites Carcinoma (EAC) cell line.Methods: The antitumor activity of the chloroform and ethanol extracts of Cuscuta reflexa was evaluated against Ehrlich ascites carcinoma (EAC) tumor in mice at doses of 200 and 400 mg/kg body weight orally, respectively, while acute oral toxicity studies were performed to determine the safety of the extracts. Briefly, the EAC cells were injected (i.p.) into ninty six mice (divided into 6 numerically equal groups), and after a one-day incubation period, the extracts were administered to the mice daily for 16 days. On day 21, six animals in each group were sacrificed for observation of antitumor activity and the remaining animals were observed to determine host the life span. Antitumor effect was determined by evaluating tumor volume, viable and nonviable tumor cell count and hematological parameters of the host. The standard antitumor used was 5-fluorouracil.Results: Administration of the extracts resulted in a significant (p < 0.05) decrease in tumor volume and viable cell count, but increased non-viable cell count and mean survival time, thereby increasing the life span of the tumor-bearing mice. Restoration of hematological parameters - red blood cells (RBC), hemoglobin, white blood cells (WBC) and lymphocyte count - to normal levels in extract-treated mice was also observed.Conclusion: The results suggest that the chloroform and ethanol extracts of C. reflexa exhibit significant antitumor activity in EAC-bearing mice that is comparable to that of the reference standard, 5-fluorouracil.Keywords: Cuscuta reflexa, Ehrlich ascites carcinoma, 5-Fluorouracil, Tumor volume, Viable cell count

Diversity of pectinolytic molds on major indian mango cultivars

The diversity of pectinolytic fungi on nine major Indian mango cultivars was studied. A total of 71 moulds belonging to 10 genera and 18 species were isolated from fruit surfaces, 49 of which showed pectinase activity. Aspergillus niger was the most frequent (30%) followed by A. fumigatus, A. flavus, A. alternata, Fusarium oxysporium, A. roseogriseum and Paecilomyces variotti. A. niger isolated from the Banganapalli cultivar from Andhra Pradesh in 2010 showed the highest pectinolytic activity. The majority of the fungi showed wide pH tolerances indicating that they could be important candidates for the production of enzymes using liquid media containing mango peel, a by-product of the mango-processing industry.Keywords: Pectinase activity; pectinolytic zone; pH, A. niger; diversity index

Environmental quality assessment of Treis Island, Nicobar, India

The physico-chemical, biological and microbiological parameters of surface seawater was studied at Tries Island during a survey in 2009. The physico-chemical parameters like silicate and inorganic phosphate concentration varied significantly from 3.53 to 4.22 and 0.05 to 0.09 μmol/L, respectively at Tries Island. The zooplankton population density ranged from 4696 to 8207 Nos./m3 and the dominant group was Copepod. The zooplankton biomass also varied significantly from 0.31 to 0.72 ml/m3. The phytoplankton density and species number also varied significantly from 1020 to 1220 Nos. /L and 25 to 31 numbers, respectively. Soleniae dominated in coastal waters of Treis Island. Chlorophyll-a, and phaeophytin concentration varied between 0.12 to 0.24 and 0.06 to 0.27 mg/m3, respectively. The chlorophyll-a concentration variation was significant. Apart from that noise level, soil and plant diversity were also investigated. Soil parameters indicated its fertile nature which is being utilized for horticulture by tribal community living in the neighbouring islands. The present study gives an account of the existing environmental quality in and around the island, providing a baseline scenario to assess the environmental impacts due to developments in the future. This study also provides comparison between populated and unpopulated coastal bay/marine ecosystems

Deferred pre-emptive switch from calcineurin inhibitor to sirolimus leads to improvement in GFR and expansion of T regulatory cell population: a randomized, controlled trial

BACKGROUND: Measures to prevent chronic calcineurin inhibitor (CNI) toxicity have included limiting exposure by switching to sirolimus (SIR). SIR may favorably influence T regulator cell (T(reg)) population. This randomized controlled trial compares the effect of switching from CNI to SIR on glomerular filtration rate (GFR) and T(reg) frequency. METHODS: In this prospective open label randomized trial, primary living donor kidney transplant recipients on CNI-based immunosuppression were randomized to continue CNI or switched to sirolimus 2 months after surgery; 29 were randomized to receive CNI and 31 to SIR. All patients received mycophenolate mofetil and steroids. The main outcome parameter was estimated GFR (eGFR) at 180 days. T(reg) population was estimated by flowcytometry. RESULTS: Baseline characteristics in the two groups were similar. Forty-eight patients completed the trial. At six months, patients in the SIR group had significantly higher eGFR as compared to those in the CNI group (88.94 ± 11.78 vs 80.59 ± 16.51 mL/min, p = 0.038). Patients on SIR had a 12 mL/min gain of eGFR of at the end of six months. Patients in the SIR group showed significant increase in T(reg) population at 30 days, which persisted till day 180. There was no difference in the adverse events in terms of number of acute rejection episodes, death, infections, proteinuria, lipid profile, blood pressure control and hematological parameters between the two groups. Four patients taking SIR developed enthesitis. No patient left the study or switched treatment because of adverse event. CONCLUSIONS: A deferred pre-emptive switch over from CNI to SIR safely improves renal function and T(reg) population at 6 months in living donor kidney transplant recipients. Registered in Clinical Trials Registry of India (CTRI/2011/091/000034)

Cholecalciferol supplementation and angiogenic markers in chronic kidney disease

Vitamin D plays an important role in proliferation and differentiation of cells and deficiency of vitamin D disturbs angiogenic balance. Previous studies in animal models have reported an association between serum levels of vitamin D and balance between pro- and anti-angiogenic factors. There is insufficient evidence about the effect of vitamin D on mediators of angiogenesis in patients with CKD. We investigated the effect of cholecalciferol supplementation on serum levels of angiogenic markers in non-diabetic patients with CKD stage 3-4. In this secondary analysis on stored samples of our previously published randomized, double-blind, placebo-controlled trial, stable patients of either sex, aged 18-70 years, with non-diabetic CKD stage 3-4 and vitamin D deficiency (serum 25-hydroxyvitamin D ≤20 ng/ml) were randomized to receive either two directly observed oral doses of cholecalciferol (300,000 IU) or matching placebo at baseline and 8 weeks. The primary outcome was change in brachial artery flow-mediated dilatation at 16 weeks. Changes in levels of serum angiogenesis markers (angiopoietin-1, angiopoietin-2, VEGF-A, VEGEF-R, and Tie-2) between groups over 16 weeks were compared. A total 120 patients were enrolled. Supplementation with cholecalciferol led to significant improvement in FMD. Serum 25(OH)D levels were similar in both groups at baseline (13.21±4.78 ng/ml and 13.40±4.42 ng/ml; p = 0.888). At 16 weeks, the serum 25(OH)D levels increased in the cholecalciferol group but not in the placebo group (between-group difference in mean change:23.40 ng/ml; 95% CI, 19.76 to 27.06; p<0.001). Serum levels of angiogenic markers were similar at baseline. At 16 weeks, angiopoietin-2 level decreased in cholecalciferol group (mean difference:-0.73 ng/ml, 95%CI, -1.25 to -0.20, p = 0.002) but not in placebo group (mean difference -0.46 ng/ml, 95%CI, -1.09 to 0.17, p = 0.154), however there was no between-group difference at 16 weeks (between-group difference in mean change: -0.27 ng/ml, 95%CI, -1.09 to 0.55, p = 0.624). Serum angiopoietin-1 level increased [mean change: 5.63 (0.51 to 10.75), p = 0.018] and VEGF-R level decreased [mean change: -87.16 (-131.89 to -42.44), p<0.001] in placebo group but did not show any change in cholecalciferol group. Our data shows the changes in Ang-1, Ang-2 and Ang-1/Ang-2 ratio after high dose oral cholecalciferol supplementation in patients with non-diabetic G3-4 CKD. The data suggests changes in circulating levels of angiogenic markers which needs to be confirmed through an adequately powered study

Alu-miRNA interactions modulate transcript isoform diversity in stress response and reveal signatures of positive selection

Primate-specific Alus harbor different regulatory features, including miRNA targets. In this study, we provide evidence for miRNA-mediated modulation of transcript isoform levels during heat-shock response through exaptation of Alu-miRNA sites in mature mRNA. We performed genome-wide expression profiling coupled with functional validation of miRNA target sites within exonized Alus, and analyzed conservation of these targets across primates. We observed that two miRNAs (miR-15a-3p and miR-302d-3p) elevated in stress response, target RAD1, GTSE1, NR2C1, FKBP9 and UBE2I exclusively within Alu. These genes map onto the p53 regulatory network. Ectopic overexpression of miR-15a-3p downregulates GTSE1 and RAD1 at the protein level and enhances cell survival. This Alu-mediated fine-tuning seems to be unique to humans as evident from the absence of orthologous sites in other primate lineages. We further analyzed signatures of selection on Alu-miRNA targets in the genome, using 1000 Genomes Phase-I data. We found that 198 out of 3177 Alu-exonized genes exhibit signatures of selection within Alu-miRNA sites, with 60 of them containing SNPs supported by multiple evidences (global-FST > 0.3, pair-wise-FST > 0.5, Fay-Wu's H  2.0, high ΔDAF) and implicated in p53 network. We propose that by affecting multiple genes, Alu-miRNA interactions have the potential to facilitate population-level adaptations in response to environmental challenges

Vitamin D deficiency, endothelial function and bone biomarkers in post-kidney transplantation patients from North India.

PURPOSE: CKD patients after kidney transplantation continue to suffer from elevated CV events which may be related to low vitamin D and its adverse impact on vascular function. The prevalence of vitamin D deficiency in North Indian kidney transplantation patients and its impact on vascular and bone biomarkers is unknown which this study investigated. METHODS: Non-diabetic, stable, > 6 months post-kidney transplantation patients, not on vitamin D supplementation, were recruited after informed consent. Data on demographics, anthropometrics and treatment were collected. Blood samples were stored at - 80 °C until analysis for bone and endothelial cell biomarkers using standard ELISA techniques. RESULTS: The clinical characteristics were: age 37.4 ± 9.9 years, 80% men, 27% ex-smokers, BP 125.5 ± 15.7/78.6 ± 9.7 mmHg, cholesterol 172.0 ± 47.8 mg/dL, hemoglobin 12.6 ± 2.3 g/dL, calcium 9.5 ± 0.6 mg/d and iPTH 58.4 ± 32.9 ng/mL and vitamin D 36.5 ± 39.8 nmol/L. Patients with vitamin D < 37.5 nmol/L (66%) had similar age, serum creatinine, serum phosphate, iPTH, blood pressure but lower calcium (9.3 ± 0.7 vs. 9.6 ± 0.5 mg/dL; p = 0.024), lower FGF23 (median 18.8 vs. 80.0 pg/mL; p = 0.013) and higher E-selectin (15.8 ± 7.9 vs. 13.0 ± 5.5 ng/mL; p = 0.047). On Univariate analysis, E-selectin (r = - 0.292; p = 0.005), FGF23 (r = 0.217; p = 0.036) and calcium (r = 0.238; p = 0.022) were significantly correlated with vitamin D levels. On stepwise multiple regression analysis, only E-selectin was associated with vitamin D levels (β = - 0.324; p = 0.002). CONCLUSION: Vitamin D deficiency was common in kidney transplant recipients in North India, associated with low FGF23 and high E-selectin. These findings suggest further investigations to assess the role of vitamin D deficiency-associated endothelial dysfunction, its implications and reversibility in kidney transplantation recipients

Effect of vitamin D supplementation on serum sclerostin levels in chronic kidney disease.

Vitamin D deficiency, cardiovascular disease and abnormal bone mineral metabolism are common in chronic kidney disease (CKD). Abnormal bone mineral metabolism has been linked to vascular calcification in CKD. Sclerostin has emerged as an important messenger in cross talk between bone-vascular axis. We analyzed sclerostin in subjects who participated in the randomized, double blind, placebo controlled trial investigating the effect of cholecalciferol supplementation on vascular function in non-diabetic CKD stage G3-4 and vitamin D ≤20ng/ml [CTRI/2013/05/003648]. Patients were randomized (1:1) to receive either two directly observed oral doses of 300,000 IU of cholecalciferol or matching placebo at baseline and 8 weeks. Of the 120 subjects enrolled, 58 in the cholecalciferol group and 59 in the placebo group completed the study. At baseline, serum levels of sclerostin were similar in both groups (cholecalciferol - median;190pg/ml, IQR;140-260pg/ml and placebo - median;180pg/ml, IQR; 140-240pg/ml, p=0.67). 16 weeks after cholecalciferol supplementation, there was no change in level of sclerostin (mean change;1.10 pg/ml, 95%CI; -27.34 to 29.34 pg/ml, p=0.25). However, a significant decrease in sclerostin level was noted in the placebo group (mean change; -31.94pg/ml, 95%CI; -54.76 to -9.13 pg/ml, p=0.002). Change (Δ) in sclerostin level at 16 weeks correlated negatively with Δ eGFR (r=-0.20, p=0.03) and positively with Δuric acid (r=0.37, p<0.001) but not with Δ25(OH) D (r=0.06, p=0.54), Δ iPTH (r=-0.03, p=0.78) ΔFGF23 (r=-0.08, p=0.38) and Δ1,25 (OH)2 D (r=-0.04, p=0.65). In conclusion, high dose cholecalciferol supplementation did not change sclerostin levels in non-diabetic stage 3-4 CKD subjects