Variations in physico-chemical and antioxidant attributes of grape seed oil as function of extraction techniques

The aim of the current research work was to assess and compare the impact of two extraction techniques on the physico-chemical, and antioxidant parameters of grape seed oil (GSO). The GSO extracted by Soxhlet and Folch methods indicated a notable variation in the oil yield (8.58 % and 10.19%) and saponification value (196.35 and 189.33 mg of KOH g-1), respectively. However, no significant (p<0.05) variation was detected for density, acid value, refractive index, iodine no, unsaponifiable matter, and free fatty acids between the tested two oils. Meanwhile, the oil produced by Soxhlet method exhibited relatively a higher extent of unsaturated dienes, trienes, peroxide value, p-anisidine value, and thus poor oxidation state. A notable variation in the content of principal fatty acid (linoleic acid) was recorded between Folch extracted oil (70.11%) and Soxhlet extracted oil (66.57%). The contents of total tocopherols were noted to be considerably higher for Folch extracted oil (105.55 mg kg-1) than the Soxhlet extracted oil (73.70 mg kg-1). Among the individual phenolics analyzed by HPLC, gallic acid (14.02 mg kg-1) and caffeic acid (5.20 mg kg-1) were detected as major component in Folch extracted oil and Soxhlet extracted oil, respectively. The results of the present comparative study support that Folch method is relatively a good choice for the extraction of GSO with promising nutritive quality in terms of oxidation parameters, contents of linoleic acid and antioxidant phenolics

Mental Health Prevention and Promotion in General Practice Settings: A Protocol for a Feasibility Study

Background A reactive approach is typically taken when addressing and intervening with mental health problems rather than a proactive or preventative one, yet preventative approaches can also reduce mental ill-health. This study protocol aims to evaluate the feasibility of recruiting general practice patients into a randomised feasibility study where they will receive either mental health treatment as usual or a brief psychological intervention for preventing the deterioration of mental health and promoting emotional wellbeing. Methods This is a two-arm RCT, where participants will be randomised to either: treatment-as-usual within GP; or treatment-as-usual within GP plus a mental health prevention and promotion intervention. Sixty patients, aged 16+ from GP surgeries, with mild to moderate mental health difficulties as indicated by the PHQ9 and GAD7 will be recruited. Data on engagement with the intervention will be summarised using descriptive statistics. Regression models will be fitted, using the 12-week post-intervention follow-up data as the outcome variable and age, gender, trial arm and the corresponding baseline data as covariates. Cost-effectiveness will be investigated in an explorative way. Descriptive statistics will be used to analyse participant's resource use and HRQoL. Qualitative data will understand factors that facilitate or challenge the successful implementation of interventions and a process evaluation will provide insight into the intervention's mechanisms of action. Discussion The research team will progress from a feasibility RCT to a larger definitive RCT and disseminate widely across stakeholders (clinical, academic, service users, caregivers, Integrated Care Board (ICB) colleagues), ensuring accessibility in collaboration with the PPI committee

Imaging the NADH:NAD+ homeostasis for understanding the metabolic response of Mycobacterium to physiologically relevant stresses

The NADH:NAD+ ratio is the primary indicator of the metabolic state of bacteria. NAD(H) homeostasis is critical for Mycobacterium tuberculosis (Mtb) survival and is thus considered an important drug target, but the spatio-temporal measurements of NAD(H) remain a challenge. Genetically encoded fluorescent biosensors of the NADH:NAD+ ratios were recently described, paving the way for investigations of the metabolic state of pathogens during infection. Here we have adapted the genetically encoded biosensor Peredox for measurement of the metabolic state of Mtb in vitro and during infection of macrophage cells. Using Peredox, here we show that inhibition of the electron transport chain, disruption of the membrane potential and proton gradient, exposure to reactive oxygen species and treatment with antimycobacterial drugs led to the accumulation of NADH in mycobacterial cells. We have further demonstrated that Mtb residing in macrophages displays higher NADH:NAD+ ratios, that may indicate a metabolic stress faced by the intracellular Mtb. We also demonstrate that the Mtb residing in macrophages display a metabolic heterogeneity, which may perhaps explain the tolerance displayed by intracellular Mtb. Next we studied the effect of immunological modulation by interferon gamma on metabolism of intracellular Mtb, since macrophage activation is known to restrict mycobacterial growth. We observed that activation of resting macrophages with interferon-gamma results in higher NADH:NAD+ levels in resident Mtb cells. We have further demonstrated that exposure of Isoniazid, Bedaquiline, Rifampicin, and O-floxacin results in higher NADH:NAD+ ratios in the Mtb residing in macrophages. However intracellular Mtb displays lower NADH:NAD+ ratio upon exposure to clofazimine. In summary, we have generated reporter strains capable of measuring the metabolic state of Mtb cells in vitro and in vivo with spatio-temporal resolution. We believe that this tool will facilitate further studies on mycobacterial physiology and will create new avenues of research for anti-tuberculosis drug discovery

Frequency of Gallstones in Fatty Liver disease in a Tertiary Care of Suburban Islamabad

Background: Gallstone disease and Non-Alcoholic Fatty Liver Disease (NAFLD) share common etiological pathways. The objective of this study was to determine the frequency of Gallstones and Non-Alcoholic Fatty Liver disease (NAFLD) in patients undergoing abdominal ultrasounds in our hospital and to compare the frequency of Gallstone disease in patients with NAFLD and without NAFLD. Methodology: This cross-sectional study was carried out at HBS General Hospital, Islamabad from January 2020 till December 2020. Patients undergoing routine ultrasounds in the out-patient department of the hospital were included in the study. Patients having Emergency Ultrasounds, history of Alcohol intake, history of chronic liver disease and pregnant patients were excluded from the study.&nbsp; Data was analysed using Microsoft Excel 2016 and IBM SPSS 22. Results: A total of 689 patients were included with a 3:1 Female to Male ratio. Mean Age was 40.65± 15.610 years. The prevalence of Gallstone disease was found to be 20.9% while that of NAFLD 27.6%. A higher percentage of patients with NAFLD had Gallstones as compared to those who did not have NAFLD (24.2% vs 19.6% respectively) however for overall data, statistical significance was not reached. A statistically significant greater number of young patients with NAFLD had Gallstones as compared to those who did not have NAFLD (50% vs 6%, p&lt;0.001). Mean age was greater in patients who had NAFLD as well as those having Gallstones. Conclusion: The frequency of Gallstones was greater in patients with NAFLD as compared to those without NAFLD. Keywords: Cholelithiasis, Fatty Liver Disease,&nbsp; Gallstones, Non-Alcoholic, Ultrasonogram &nbsp

Glycated haemoglobin: A marker of circulating lipids in patients with type 2 diabetes

Introduction: Type 2 diabetes mellitus (T2DM) is a group of metabolic disorder and is an independent risk factor for cardiovascular disease and dyslipidaemia. Patients with T2DM have dyslipidaemia at wavering degrees, characterised by increased levels of triglyceride and low-density lipoprotein-cholesterol and decreased levels of high-density lipoprotein (HDL)-cholesterol. In the present study, we evaluated glycated haemoglobin (HbA1c) as a marker of circulating lipids in patients with T2DM. Methods: Two hundred and thirty-nine patients with T2DM were enrolled for the study. A detailed biochemical and lipid profile was done for all patients. Results: Of 239 cases, 96 (40%) were male and 143 (60%) were female. Of 239 patients, 53 (22%) patients with T2DM had controlled glycaemia (HbA1c <6.5) and 186 (78%) patients had uncontrolled glycaemia (HbA1c ≥6.5). Pearson's correlation of HbA1c with all lipid parameters was statistically significant. HbA1c, however, had an inverse correlation with HDL and had a significant direct correlation with fasting blood glucose. Conclusion: The study reveals that HbA1c is not only a reliable glycaemic index but can also be used as an important indicator of dyslipidaemia in patients with T2DM

MICROBIOLOGICAL STUDIES OF BOVINE MASTITIS CAUSED BY ESCHERICHIA COLI IN QUETTA, BALOCHISTAN

Objective: The following study aims at determine the prevalence of bacterial mastitis in dairy cows in Quetta, to isolate and identify E.coli from dairy cows in Quetta, characterize biochemically and test for antibiotic sensitivity along with DNA extraction and confirmation of E.coli via PCR. Methods and Materials: The milk samples were collected from different government and private dairy farms of Quetta city. The samples were streaked on MacConkey agar and kept in incubator at 37 ̊C for 24 hours. Antibiotic sensitivity test was performed by using disc diffusion Bauer technique and McFarland Turbidity Standard method 0.5 following CLSI protocols. The products were separated with 1% agarose gel electrophoresis and stained with ethidium bromide and images were documented during PCR. Results: The overall prevalence of bovine mastitis among cows and buffalos was 38% with 18 % in government and 8% in private dairy farms. Antibiotics result showed that resistant to Vancomycin ,Lincomycin , Carbenicillin, Kanamycin. The PCR amplification was positive for our isolation. Conclusion: Present study concluded that E.coli is responsible for the high rate of mastitis among cows and buffalos in Quetta city. The E.coli found in the dairy farms of the Quetta was found to be resistant to the five antibiotics. This is an alarming state indicating the rising antibiotic resistance of E.coli towards the various antibiotic drug classes. The lack of awareness of the proper cleanliness and hygiene measures at dairy farms could be denoted as the reason of the higher prevalence of the mastitis in the dairy farms of Quetta

Purification and characterization of a cold active alkaline protease from Stenotrophomonas sp., isolated from Kashmir, India

A Psychrotolerant alkaline protease producing bacterium IIIM-ST045 was isolated from a soil sample collected from the Thajiwas glacier of Kashmir, India and identified as Stenotrophomonas sp. on the basis of its biochemical properties and 16S ribosomal gene sequencing. The strain could grow well within a temperature range of 4–37�C however, showed optimum growth at 15�C. The strain was found to over-produce proteases when it was grown in media containing lactose as carbon source (157.50 U mg-1). The maximum specific enzyme activity (398 U mg-1) was obtained using soya oil as nitrogen source, however, the inorganic nitrogen sources urea,ammonium chloride and ammonium sulphate showed the lowest production of 38.9, 62.2 and 57.9 U mg-1. The enzyme was purified to 18.45 folds and the molecular weight of the partially purified protease was estimated to be *55 kDa by SDS-PAGE analysis. The protease activity increased as the increase in enzyme concentration while as the optimum enzyme activity was found when casein (1% w/v) was used as substrate. The enzyme was highly active over a wide range of pH from 6.5 to 12.0 showing optimum activity at pH 10.0. The optimum temperature for the enzyme was 15�C. Proteolytic activity reduced gradually with higher temperatures with a decrease of 56% at 40�C. The purified enzyme was checked for the removal of protein containing tea stains using a silk cloth within a temperature range of 10–60�C. The best washing efficiency results obtained at low temperatures indicate that the enzyme may be used for cold washing purposes of delicate fabrics that otherwise are vulnerable to high temperatures

Prevalence of metabolic syndrome in the family members of women with polycystic ovary syndrome from North India

Background: Polycystic ovary syndrome (PCOS) is the most complex and common endocrine disorder of women in reproductive years. In addition to irregular menstrual cycles, chronic anovulation and hyperandrogenism, it has many metabolic manifestations such as obesity, hyperlipidemia, hyperinsulinemia, insulin resistance, dysglycemia, increased risk of cardiovascular disease or possibly endometrial cancer. Familial clustering of PCOS in consistence with the genetic susceptibility has been described. Materials and Methods: The present study assessed the clinical, biochemical and hormonal parameters including prevalence of metabolic syndrome by two different criteria in the first- degree relatives of patients with PCOS. Results: The average age of 37 index patients was 23 ± 3.6 years, with the mean age of menarche as 13.3 ± 1.2 years. The mean age and age of menarche in mothers (n = 22) was 48.8 ± 5.1 and 13 ± 1.3 years, respectively, whereas as it was 23.5 ± 4.7 and 13.3 ± 1.2 years in sisters (n = 22), respectively. Metabolic syndrome (MS) defined by International Diabetes Federation (IDF) criteria was present in 10 index patients, 1 brother, 4 sisters, 17 mothers and 15 fathers while as by Adult Treatment Panel III (ATP III) it was in 8 index patients, 5 sisters, 16 mothers and 11 fathers. Conclusion: The presence of MS or related metabolic derangements is high in the family members of women with PCOS