Application of glucose oxidase for the production of metal gluconates by fermentation

The present study deals with the application of glucose oxidase (GOX) for the production of metal gluconates by fermentation method. It provides a method for the conversion of glucose into gluconic acid and its derivatives using the enzyme glucose oxidase (GOX). Due to the presence of calcium carbonate in fermentation medium the gluconic acid is converted into calcium gluconate. Conditions like concentration of substrate, temperature, pH, fermentation period and different phosphate sources were optimized during fermentation. The maximum GOX activity was observed at 35°C (pH 5.5) after 44 h of incubation at 100 rpm. At the maximum enzyme activity, the percentage yield of gluconates are also maximum; both go side by side. Sulphuric and oxalic acids method were employed for the production of gluconic acid. Derivatives of gluconic acid that is, calcium lactate gluconate, sodium gluconate, potassium gluconate, zinc gluconate and copper gluconate were formed by using double displacement and direct methods. The direct method gave the better yield. The percentage yields were 73, 89.63, 81.93, 92.86 and 81.53%, respectively. Keywords: Glucose oxidase (GOX), metal gluconate, double displacement

Analytical study of reaction diffusion Lengyel-Epstein system by generalized Riccati equation mapping method

In this study, the Lengyel-Epstein system is under investigation analytically. This is the reaction–diffusion system leading to the concentration of the inhibitor chlorite and the activator iodide, respectively. These concentrations of the inhibitor chlorite and the activator iodide are shown in the form of wave solutions. This is a reaction†“diffusion model which considered for the first time analytically to explore the different abundant families of solitary wave structures. These exact solitary wave solutions are obtained by applying the generalized Riccati equation mapping method. The single and combined wave solutions are observed in shock, complex solitary-shock, shock singular, and periodic-singular forms. The rational solutions also emerged during the derivation. In the Lengyel-Epstein system, solitary waves can propagate at various rates. The harmony of the system’s diffusive and reactive effects frequently governs the speed of a single wave. Solitary waves can move at a variety of speeds depending on the factors and reaction kinetics. To show their physical behavior, the 3D and their corresponding contour plots are drawn for the different values of constants

A mechanistic insight into chemical cues and interactions involved in herbivory induced jasmonate mediated plant defense mechanism

The first step in plant defense mechanism is to sense the insect attack stimulus. Plant sensitivity of an insect attack is the first step of defense. Molecules generated by the oral secretion of the insect interact with the plant receptors to trigger plant defense mechanisms. We selected some highly cited insect elicitors molecules, volicitin, caeliferin, bruchin which interact with plant defense by interacting with plant elicitors (systemin, inceptin and peps) located on the plant cell surface. This interaction activates plant receptors SYR1, LRR, PEPR and triggers downstream defense signaling. The octadecanoid pathways, involving enzymes allene oxide synthase (AOS) and Hydroxyperoxide lyase (HPL) are activated. These enzymes mediate production of green leafy volatiles and Jasmonic acid by interacting with hydroxperoxide molecules. We docked the elicitors with receptors and enzymes with substrates in the pathway of JA production. Phe was found to be an important amino acid that interacts with 13-hydroxyperoxides in the case of AOS to produce JA but not in the case of HPL. JA is converted to JA-Ile which shows strong binding with COI1 and COI1-JA-Ile complex docked with JAZ which showed strong interaction with ve hydrogens and one salt bridge bond. AOS and HPL showed less than 40% identity for sequence and structure alignment. AOS and HPL had shown an interaction between each other and showed a common interaction partner of the Lipoxygenase family. HPL shows interaction with ADH2 (Alcohol dehydrogenase) involved in GLVs production. AOS also showed interaction partner AOC, COI1 and OPR1 which are involved in JA-induced plant defense mechanism.peer-reviewe

Effects of antibiotic resistance, drug target attainment, bacterial pathogenicity and virulence, and antibiotic access and affordability on outcomes in neonatal sepsis: an international microbiology and drug evaluation prospective substudy (BARNARDS)

Background Sepsis is a major contributor to neonatal mortality, particularly in low-income and middle-income countries (LMICs). WHO advocates ampicillin–gentamicin as first-line therapy for the management of neonatal sepsis. In the BARNARDS observational cohort study of neonatal sepsis and antimicrobial resistance in LMICs, common sepsis pathogens were characterised via whole genome sequencing (WGS) and antimicrobial resistance profiles. In this substudy of BARNARDS, we aimed to assess the use and efficacy of empirical antibiotic therapies commonly used in LMICs for neonatal sepsis. Methods In BARNARDS, consenting mother–neonates aged 0–60 days dyads were enrolled on delivery or neonatal presentation with suspected sepsis at 12 BARNARDS clinical sites in Bangladesh, Ethiopia, India, Pakistan, Nigeria, Rwanda, and South Africa. Stillborn babies were excluded from the study. Blood samples were collected from neonates presenting with clinical signs of sepsis, and WGS and minimum inhibitory concentrations for antibiotic treatment were determined for bacterial isolates from culture-confirmed sepsis. Neonatal outcome data were collected following enrolment until 60 days of life. Antibiotic usage and neonatal outcome data were assessed. Survival analyses were adjusted to take into account potential clinical confounding variables related to the birth and pathogen. Additionally, resistance profiles, pharmacokinetic–pharmacodynamic probability of target attainment, and frequency of resistance (ie, resistance defined by in-vitro growth of isolates when challenged by antibiotics) were assessed. Questionnaires on health structures and antibiotic costs evaluated accessibility and affordability. Findings Between Nov 12, 2015, and Feb 1, 2018, 36 285 neonates were enrolled into the main BARNARDS study, of whom 9874 had clinically diagnosed sepsis and 5749 had available antibiotic data. The four most commonly prescribed antibiotic combinations given to 4451 neonates (77·42%) of 5749 were ampicillin–gentamicin, ceftazidime–amikacin, piperacillin–tazobactam–amikacin, and amoxicillin clavulanate–amikacin. This dataset assessed 476 prescriptions for 442 neonates treated with one of these antibiotic combinations with WGS data (all BARNARDS countries were represented in this subset except India). Multiple pathogens were isolated, totalling 457 isolates. Reported mortality was lower for neonates treated with ceftazidime–amikacin than for neonates treated with ampicillin–gentamicin (hazard ratio [adjusted for clinical variables considered potential confounders to outcomes] 0·32, 95% CI 0·14–0·72; p=0·0060). Of 390 Gram-negative isolates, 379 (97·2%) were resistant to ampicillin and 274 (70·3%) were resistant to gentamicin. Susceptibility of Gram-negative isolates to at least one antibiotic in a treatment combination was noted in 111 (28·5%) to ampicillin–gentamicin; 286 (73·3%) to amoxicillin clavulanate–amikacin; 301 (77·2%) to ceftazidime–amikacin; and 312 (80·0%) to piperacillin–tazobactam–amikacin. A probability of target attainment of 80% or more was noted in 26 neonates (33·7% [SD 0·59]) of 78 with ampicillin–gentamicin; 15 (68·0% [3·84]) of 27 with amoxicillin clavulanate–amikacin; 93 (92·7% [0·24]) of 109 with ceftazidime–amikacin; and 70 (85·3% [0·47]) of 76 with piperacillin–tazobactam–amikacin. However, antibiotic and country effects could not be distinguished. Frequency of resistance was recorded most frequently with fosfomycin (in 78 isolates [68·4%] of 114), followed by colistin (55 isolates [57·3%] of 96), and gentamicin (62 isolates [53·0%] of 117). Sites in six of the seven countries (excluding South Africa) stated that the cost of antibiotics would influence treatment of neonatal sepsis

Convalescent plasma in patients admitted to hospital with COVID-19 (RECOVERY): a randomised controlled, open-label, platform trial

SummaryBackground Azithromycin has been proposed as a treatment for COVID-19 on the basis of its immunomodulatoryactions. We aimed to evaluate the safety and efficacy of azithromycin in patients admitted to hospital with COVID-19.Methods In this randomised, controlled, open-label, adaptive platform trial (Randomised Evaluation of COVID-19Therapy [RECOVERY]), several possible treatments were compared with usual care in patients admitted to hospitalwith COVID-19 in the UK. The trial is underway at 176 hospitals in the UK. Eligible and consenting patients wererandomly allocated to either usual standard of care alone or usual standard of care plus azithromycin 500 mg once perday by mouth or intravenously for 10 days or until discharge (or allocation to one of the other RECOVERY treatmentgroups). Patients were assigned via web-based simple (unstratified) randomisation with allocation concealment andwere twice as likely to be randomly assigned to usual care than to any of the active treatment groups. Participants andlocal study staff were not masked to the allocated treatment, but all others involved in the trial were masked to theoutcome data during the trial. The primary outcome was 28-day all-cause mortality, assessed in the intention-to-treatpopulation. The trial is registered with ISRCTN, 50189673, and ClinicalTrials.gov, NCT04381936.Findings Between April 7 and Nov 27, 2020, of 16 442 patients enrolled in the RECOVERY trial, 9433 (57%) wereeligible and 7763 were included in the assessment of azithromycin. The mean age of these study participants was65·3 years (SD 15·7) and approximately a third were women (2944 [38%] of 7763). 2582 patients were randomlyallocated to receive azithromycin and 5181 patients were randomly allocated to usual care alone. Overall,561 (22%) patients allocated to azithromycin and 1162 (22%) patients allocated to usual care died within 28 days(rate ratio 0·97, 95% CI 0·87–1·07; p=0·50). No significant difference was seen in duration of hospital stay (median10 days [IQR 5 to >28] vs 11 days [5 to >28]) or the proportion of patients discharged from hospital alive within 28 days(rate ratio 1·04, 95% CI 0·98–1·10; p=0·19). Among those not on invasive mechanical ventilation at baseline, nosignificant difference was seen in the proportion meeting the composite endpoint of invasive mechanical ventilationor death (risk ratio 0·95, 95% CI 0·87–1·03; p=0·24).Interpretation In patients admitted to hospital with COVID-19, azithromycin did not improve survival or otherprespecified clinical outcomes. Azithromycin use in patients admitted to hospital with COVID-19 should be restrictedto patients in whom there is a clear antimicrobial indication

EVALUATION OF OXIDATIVE STRESS INDUCED HEPATOTOXICITY PRODUCED BY CISPLATIN IN MALE SPRAGUE DAWLEY RATS

Objective: To study the effect of cisplatin administration on oxidative stress induced hepatotoxicity in male Sprague Dawley rats. Study Design: Randomized controlled trial. Place and Duration of Study: Study was conducted at department of Physiology, Army Medical College, Rawalpindi. Duration of study was 18 months from Oct 2014 to Apr 2016. Material and Methods: The trial was performed on sixty male Sprague Dawley rats which were distributed randomly into two groups of 30 rats each. Group I received placebo whereas group II received intraperitoneal cisplatin 2mg/kg body weight two times a week for the period of 4 weeks. After successful treatment, animals were sacrificed and terminal blood sample was collected and used for estimation of serum AST, ALT, albumin and 8-isoprostane. Dissection of rats were done and liver tissue sampled. Tissue homogenate was prepared from liver sample which was used for estimation of total glutathione levels. Results: In group I, 8-isoprostane was 18.31 ± 3.35 pg/ml, total glutathione was 4.29 ± 0.42 μmol/L, ALT was 36.93 ± 4.72 IU/L, AST was 124.2 ± 12.75 IU/L and Albumin was 4.11 ± 0.26 g/dl whereas in group II, 8-isoprostane was 67.9 ± 8.14 pg/ml, total glutathione was 1.92 ± 0.28 μmol/L, ALT was 87.17 ± 6.47 IU/L, AST was 357.7 ± 19.37 IU/L and Albumin was 2.12 ± 0.25 g/dl. Levels of serum 8-isoprostane, ALT and AST were significantly raised (p<0.001) whereas serum albumin and total glutathione in liver tissue were found significantly low (p<0.001) in group II as compared to group I. Conclusion: Cisplatin treatment causes hepatotoxicity by increased production of reactive oxygen species in male Sprague Dawley rats

Decoding type 2 diabetes mellitus genetic risk variants in Pakistani Pashtun ethnic population using the nascent whole exome sequencing and MassARRAY genotyping: A case-control association study.

Genome-wide association studies have greatly increased the number of T2DM associated risk variants but most of them have focused on populations of European origin. There is scarcity of such studies in developing countries including Pakistan. High prevalence of T2DM in Pakistani population prompted us to design this study. We have devised a two stage (the discovery stage and validation stage) case-control study in Pashtun ethnic population in which 500 T2DM cases and controls each have been recruited to investigate T2DM genetic risk variants. In discovery stage Whole Exome Sequencing (WES) was used to identify and suggest T2DM pathogenic SNPs, based on SIFT and Polyphen scores; whereas in validation stage the selected variants were confirmed for T2DM association using MassARRAY genotyping and appropriate statistical tests. Results of the study showed the target positive association of rs1801282/PPARG (OR = 1.24, 95%Cl = 1.20-1.46, P = 0.010), rs745975/HNF4A (OR = 1.30, 95%Cl = 1.06-1.38, P = 0.004), rs806052/GLIS3 (OR = 1.32, 95%Cl = 1.07-1.66, P = 0.016), rs8192552/MTNR1B (OR = 1.53, 95%Cl = 0.56-1.95, P = 0.012) and rs1805097/IRS-2 (OR = 1.27, 95%Cl = 1.36-1.92, P = 0.045), with T2DM; whereas rs6415788/GLIS3, rs61788900/NOTCH2, rs61788901/NOTCH2 and rs11810554/NOTCH2 (P>0.05) showed no significant association. Identification of genetic risk factors/variants can be used in defining high risk subjects assessment, and disease prevention

Supplementation of PUFA extracted from microalgae for the development of chicken patties

In recent years, there has been a growing interest in development of a diverse range of foods that are enriched with omega-3 fatty acids. It is widely recognized that through dietary interventions, the lipid fraction of food can be modified to enhance its nutritional content. This study is aimed to develop chicken patties enriched with poly unstaurated fatty acids (PUFAs) extracted from microalgae aurintricarboxylic acid (ATA) concentration of 0% (T0), 1% (T1), 2% (T2), and 3% (T3). All treatments were stored at −18 °C for one month and analysed at an interval of 0, 10, 20, and 30 days to assess the effect of PUFAs supplementation on physicochemical, oxidative, microbiological and organoleptic properties of chicken patties. The results revealed that moisture content was significantly increased during the storage; the maximum moisture was observed in T0 (67.25% ± 0.03) on day 0, while the minimun was found in T3 (64.69% ± 0.04) on day 30. Supplemenatation of PUFAs in chicken patties significantly enhanced the fat content of the product the highest fat content was observed for T3 (9.7% ± 0.06. An increase in PUFAs concentration led to a significant increase in thiobarbituric acid reactive substances (TBARS). TBARS were increased from 1.22 ± 0.43 at 0 days to 1.48 ± 0.39 at 30 days of storage. The PUFAs incorporation negatively effected sensory acceptance of the product ranging from (8.41 ± 0.17 to 7.28 ± 0.12). However, the sensory scores were in acceptable range for supplemented patties as compared to control sample. Treatment T3 depicted the highest nutritional content. The sensory and physiochemical analysis of supplemented patties suggested that PUFAs extracted from microalgae can be used as a functional ingredient in the preparation various meat products particularly chicken meta patties. However, antioxidants should be added to to prevent lipid oxidation in the product