Kinetic and sequence-structure-function analysis of known LinA variants with different hexachlorocyclohexane isomers

BACKGROUND Here we report specific activities of all seven naturally occurring LinA variants towards three different isomers, α, γ and δ, of a priority persistent pollutant, hexachlorocyclohexane (HCH). Sequence-structure-function differences contributing to the differences in their stereospecificity for α-, γ-, and δ-HCH and enantiospecificity for (+)- and (-)-α -HCH are also discussed. METHODOLOGY/PRINCIPAL FINDINGS Enzyme kinetic studies were performed with purified LinA variants. Models of LinA2(B90A) A110T, A111C, A110T/A111C and LinA1(B90A) were constructed using the FoldX computer algorithm. Turnover rates (min(-1)) showed that the LinAs exhibited differential substrate affinity amongst the four HCH isomers tested. α-HCH was found to be the most preferred substrate by all LinA's, followed by the γ and then δ isomer. CONCLUSIONS/SIGNIFICANCE The kinetic observations suggest that LinA-γ1-7 is the best variant for developing an enzyme-based bioremediation technology for HCH. The majority of the sequence variation in the various linA genes that have been isolated is not neutral, but alters the enantio- and stereoselectivity of the encoded proteins.This work was supported by the Indo-Australian Biotechnology Fund from the Department of Education Science and Technology (DEST), Australia and the Department of Biotechnology (DBT), India

ANCIENT METHODS OF RESEARCH IN AYURVEDA

Ayurveda is the Upaveda of Atharvaveda, the ancient literature which gave us the knowledge of healthier life. The absolute aim of Ayurveda is to maintain the health of a healthier person and diagnose and treat the diseased person. Ayurveda has the ability to treat many chronic diseases that are untreatable in modern medicine, it has got some very preventive and curative tools to treat the diseases, but unfortunately, due to the lack of scientific validation in various concepts, this precious gift from our ancestors is trailing. In a nutshell, Ayurveda gives human a healthy life with proven methods of medical treatment. The ancient methods of treatment are very effective and relevant in present time also. Hence, evidence-based research is highly needed for global recognition and acceptance of Ayurveda, which needs further advancements in the research methodology

Gall Bladder Calculus in reproductive age group females attending the ultrasound clinic at a tertiary care hospital: A comparative Cross-sectional study

Background: Pregnant females are at risk for cholelithiasis' progress, and the obstruction in the bile duct produced by gallstones is responsible for jaundice and abdominal pain. Objective: The objectives of the present study were to see the burden of gall stones, relation with parity, and trimester among reproductive age group females. Methodology: A cross-sectional study was conducted on 467 females of age between 15 to 45 years. The gall bladder was thoroughly examined by an ultrasound probe of frequency 2-5 MHz in all planes. Results: In most females who had gallstones, single (68.8 percent) gall stone was common as compared to multiple (31.2 percent). More nonpregnant women had gall bladder stones than pregnant women (p-value <0.005). Among the current nonpregnant group, the percentage of presence of gallbladder stones was greater in the female having parity of two or more (27.4 percent) than the females having parity of less than two (8.2 percent) with p-value <0.05. Conclusion: Since Gallstones can further complicate the situation in the form of gallbladder carcinoma, and females with higher parity have more chances of developing gallstones. Therefore, it becomes necessary to know the trimester of gallstone formation and its role in the progression of symptoms

MCML D-Latch Using Triple-Tail Cells: Analysis and Design

A new low-voltage MOS current mode logic (MCML) topology for D-latch is proposed. The new topology employs a triple-tail cell to lower the supply voltage requirement in comparison to traditional MCML D-latch. The design of the proposed MCML D-latch is carried out through analytical modeling of its static parameters. The delay is expressed in terms of the bias current and the voltage swing so that it can be traded off with the power consumption. The proposed low-voltage MCML D-latch is analyzed for the two design cases, namely, high-speed and power-efficient, and the performance is compared with the traditional MCML D-latch for each design case. The theoretical propositions are validated through extensive SPICE simulations using TSMC 0.18 µm CMOS technology parameters

Study of pregnancy outcome in women with cardiac disease: a retrospective analysis

Background: Cardiac disease complicates 1% of all pregnancies. It is one of the 3 major indirect causes of maternal mortality in India. Objective of the study is to evaluate the maternal and fetal outcome in patients with cardiac disease in pregnancy.Methods: A retrospective analysis was carried out in 117 pregnant women with known or newly diagnosed heart disease from January 2013 to December 2015 at a tertiary care hospital.Results: In the present study, the incidence of cardiac disease was 0.8%. Majority 66 (56.4%) were of 23-27 years of age, 72 (61.5%) were primigravida. Mostly 84 (71.8%) belonging to New York Heart Association (NYHA) Class I and II. Rheumatic heart disease seen in 96 (82.1%) and congenital heart disease in 17.9% subjects. Among RHD Mitral stenosis was the most common, seen in 51 (53.1%) cases. Majority 78 (66.6%) had vaginal delivery and caesarean section in 28 (23.9%) cases. Most common maternal complication was anaemia seen in 54 (46.2%) cases, congestive cardiac failure complicated 23 (19.6%) cases. 18 patients required ICU care and maternal mortality in 6 cases (5.12%). The live births in (94.8%) cases, 33 (28.2%) babies required NICU admission and perinatal mortality was 7.7%.Conclusions: Heart disease in pregnancy is a high risk condition and has a major impact on pregnancy outcome. Rheumatic heart disease being the prominent cardiac lesion. Fetomaternal mortality and morbidity can be reduced with proper antenatal, intrapartum and postnatal care in conjunction with cardiologist and neonatologist

Kinetic and Sequence-Structure-Function Analysis of Known LinA Variants with Different Hexachlorocyclohexane Isomers

BACKGROUND: Here we report specific activities of all seven naturally occurring LinA variants towards three different isomers, α, γ and δ, of a priority persistent pollutant, hexachlorocyclohexane (HCH). Sequence-structure-function differences contributing to the differences in their stereospecificity for α-, γ-, and δ-HCH and enantiospecificity for (+)- and (-)-α -HCH are also discussed. METHODOLOGY/PRINCIPAL FINDINGS: Enzyme kinetic studies were performed with purified LinA variants. Models of LinA2(B90A) A110T, A111C, A110T/A111C and LinA1(B90A) were constructed using the FoldX computer algorithm. Turnover rates (min(-1)) showed that the LinAs exhibited differential substrate affinity amongst the four HCH isomers tested. α-HCH was found to be the most preferred substrate by all LinA's, followed by the γ and then δ isomer. CONCLUSIONS/SIGNIFICANCE: The kinetic observations suggest that LinA-γ1-7 is the best variant for developing an enzyme-based bioremediation technology for HCH. The majority of the sequence variation in the various linA genes that have been isolated is not neutral, but alters the enantio- and stereoselectivity of the encoded proteins

Multi-omic analysis of two common p53 mutations: Proteins regulated by mutated p53 as potential targets for immunotherapy

The p53 protein is mutated in more than 50% of human cancers. Mutated p53 proteins not only lose their normal function but often acquire novel oncogenic functions, a phenomenon termed mutant p53 gain-of-function. Mutant p53 has been shown to affect the transcription of a range of genes, as well as protein–protein interactions with transcription factors and other effectors; however, no one has intensively investigated and identified these proteins, or their MHC presented epitopes, from the viewpoint of their ability to act as targets for immunotherapeutic interventions. We investigated the molecular changes that occurred after the TP53 null osteosarcoma cells, SaOS-2, were transfected with one of two conformational p53-mutants, either R175H or R273H. We then examined the phenotypic and functional changes using macroscopic observations, proliferation, gene expression and proteomics alongside immunopeptidome profiling of peptide antigen presentation in the context of major histocompatibility complex (MHC) class I molecules. We identified several candidate proteins in both TP53 mutant cell lines with differential expression when compared to the TP53 null vector control, SaOS-V. Quantitative SWATH proteomics combined with immune-peptidome analysis of the class-I eluted peptides identified several epitopes presented on pMHC and in silico analysis shortlisted which antigens were expressed in a range of cancerous but not adjacent healthy tissues. Out of all the candidates, KLC1 and TOP2A showed high levels of expression in every tumor type examined. From these proteins, three A2 and four pan HLA-A epitopes were identified in both R175H and R273H from TOP2A. We have now provided a short list of future immunotherapy targets for the treatment of cancers harboring mutated TP53

Derangement of liver function tests in dengue patients

Background: Changes in the liver function test may serve as an early marker for timely diagnosis and identification of patients who may develop severe dengue. The purpose of this study was to examine the link between dengue fever severity and liver function test. Methods: This prospective observational study was conducted in the Department of General Medicine, Madhesh Institute of health sciences, provincial hospital, Janakpurdham in which we included dengue positive patients (aged 18 years or more) based on NS1 antigen or high titer on IgM/IgG testing from July 2023 till August 2023. We excluded patients with diseases like malaria, cirrhosis of liver, enteric fever, viral hepatitis or any other disease or taking any medication which can derange LFT. Results: We included 96 patients fulfilling the study criteria. Of these, 71% had DF, 22% had DHF and 7% had DSS. Among liver enzymes, mean AST of the patients was significantly higher in DSS group of patients (775.19±39.65 U/l), as compared to those in the DF and DHF group of patients, p value <0.01. Similarly, mean ALT of the patients was significantly higher in DSS group of patients (387.8±18.6 U/l), as compared to those in the DF and DHF group of patients, p value<0.01. On the contrary, mean alkaline phosphatase levels were similar between the three patient groups. Conclusions: Based on the results our study, we conclude that raised AST and ALT levels were significantly associated with severity of DSS and DHF. Patients with dengue infection should have a baseline liver function test and subsequent LFT monitoring to detect early hepatic impairment