In silico and structure-based evaluation of deleterious mutations identified in human Chk1, Chk2, and Wee1 protein kinase.

Checkpoint kinases Chk1, Chk2, Wee1 are playing a key role in DNA damage response and genomic integrity. Cancer-associated mutations identified in human Chk1, Chk2, and Wee1 were retrieved to understand the function associated with the mutation and also alterations in the folding pattern. Therefore, an attempt has been made to identify deleterious effect of variants using in silico and structure-based approach. Variants of uncertain significance for Chk1, Chk2, and Wee1 were retrieved from different databases and four prediction servers were employed to predict pathogenicity of mutations. Further, Interpro, I-Mutant 3.0, Consurf, TM-align, and have (y)our protein explained were used for comprehensive study of the deleterious effects of variants. The sequences of Chk1, Chk2, and Wee1 were analyzed using Clustal Omega, and the three-dimensional structures of the proteins were aligned using TM-align. The molecular dynamics simulations were performed to explore the differences in folding pattern between Chk1, Chk2, Wee1 wild-type, and mutant protein and also to evaluate the structural integrity. Thirty-six variants in Chk1, 250 Variants in Chk2, and 29 in Wee1 were categorized as pathogenic using in silico prediction tools. Furthermore, 25 mutations in Chk1, 189 in Chk2, and 14 in Wee1 were highly conserved, possessing deleterious effect and also influencing the protein structure and function. These identified mutations may provide underlying genetic intricacies to serve as potential targets for therapeutic inventions and clinical management.

Time Series Modeling of Tuberculosis Cases in India from 2017 to 2022 Based on the SARIMA-NNAR Hybrid Model.

Tuberculosis (TB) is still one of the severe progressive threats in developing countries. There are some limitations to social and economic development among developing nations. The present study forecasts the notified prevalence of TB based on seasonality and trend by applying the SARIMA-NNAR hybrid model. The NIKSHAY database repository provides monthly informed TB cases (2017 to 2022) in India. A time series model was constructed based on the seasonal autoregressive integrated moving averages (SARIMA), neural network autoregressive (NNAR), and, SARIM-NNAR hybrid models. These models were estimated with the help of the Bayesian information criterion (BIC) and Akaike information criterion (AIC). These models were established to compare the estimation. A total of 12,576,746 notified TB cases were reported over the years whereas the average case was observed as 174,677.02. The evaluating parameters values of RMSE, MAE, and MAPE for the hybrid model were found to be (13738.97), (10369.48), and (06.68). SARIMA model was (19104.38), (14304.15), and (09.45) and the NNAR were (11566.83), (9049.27), and (05.37), respectively. Therefore, the NNAR model performs better with time series data for fitting and forecasting compared to other models such as SARIMA as well as the hybrid model. The NNAR model indicated a suitable model for notified TB incidence forecasting. This model can be a good tool for future prediction. This will assist in devising a policy and strategizing for better prevention and control.

Genomic analysis unveils genome degradation events and gene flux in the emergence and persistence of S. Paratyphi A lineages.

Paratyphoid fever caused by S. Paratyphi A is endemic in parts of South Asia and Southeast Asia. The proportion of enteric fever cases caused by S. Paratyphi A has substantially increased, yet only limited data is available on the population structure and genetic diversity of this serovar. We examined the phylogenetic distribution and evolutionary trajectory of S. Paratyphi A isolates collected as part of the Indian enteric fever surveillance study "Surveillance of Enteric Fever in India (SEFI)." In the study period (2017-2020), S. Paratyphi A comprised 17.6% (441/2503) of total enteric fever cases in India, with the isolates highly susceptible to all the major antibiotics used for treatment except fluoroquinolones. Phylogenetic analysis clustered the global S. Paratyphi A collection into seven lineages (A-G), and the present study isolates were distributed in lineages A, C and F. Our analysis highlights that the genome degradation events and gene acquisitions or losses are key molecular events in the evolution of new S. Paratyphi A lineages/sub-lineages. A total of 10 hypothetically disrupted coding sequences (HDCS) or pseudogenes-forming mutations possibly associated with the emergence of lineages were identified. The pan-genome analysis identified the insertion of P2/PSP3 phage and acquisition of IncX1 plasmid during the selection in 2.3.2/2.3.3 and 1.2.2 genotypes, respectively. We have identified six characteristic missense mutations associated with lipopolysaccharide (LPS) biosynthesis genes of S. Paratyphi A, however, these mutations confer only a low structural impact and possibly have minimal impact on vaccine effectiveness. Since S. Paratyphi A is human-restricted, high levels of genetic drift are not expected unless these bacteria transmit to naive hosts. However, public-health investigation and monitoring by means of genomic surveillance would be constantly needed to avoid S. Paratyphi A serovar becoming a public health threat similar to the S. Typhi of today.

Vitamin C and its therapeutic potential in the management of COVID19.

COVID19 has emerged as one of the worst pandemics in the history of mankind. Several vaccines have been approved by different government agencies worldwide, but data on their efficacy and safety are limited, and distribution remains a massive challenge. As per WHO, personal immunity is vital for protection against COVID19. Earlier, Vitamin C-mediated pathways have been shown to play critical role in boosting immunity attributed to its antioxidant properties. Recently, the involvement of such pathways in protection against COVID19 has been suggested. The controlled doses of Vitamin C administered through intravenous (IV) injections are being studied for determining its role in the prognosis of COVID19. In this article, we have discussed the potential role of Vitamin C in the management in COVID19 patients and presented recent clinical trials data. Additionally, we have elaborated the possibility of administering Vitamin C through inhalers in order to achieve local high concentration and the challenges of such approach.

Vibrio cholerae O139 genomes provide a clue to why it may have failed to usher in the eighth cholera pandemic.

Cholera is a life-threatening infectious disease that remains an important public health issue in several low and middle-income countries. In 1992, a newly identified O139 Vibrio cholerae temporarily displaced the O1 serogroup. No study has been able to answer why the potential eighth cholera pandemic (8CP) causing V. cholerae O139 emerged so successfully and then died out. We conducted a genomic study, including 330 O139 isolates, covering emergence of the serogroup in 1992 through to 2015. We noted two key genomic evolutionary changes that may have been responsible for the disappearance of genetically distinct but temporally overlapping waves (A-C) of O139. Firstly, as the waves progressed, a switch from a homogenous toxin genotype in wave-A to heterogeneous genotypes. Secondly, a gradual loss of antimicrobial resistance (AMR) with the progression of waves. We hypothesize that these two changes contributed to the eventual epidemiological decline of O139.

Microbiome Profiling of Enterotoxigenic Escherichia coli (ETEC) Carriers Highlights Signature Differences between Symptomatic and Asymptomatic Individuals.

Enterotoxigenic Escherichia coli (ETEC) is an important cause of diarrhea in children in low- and middle-income countries (LMICs). However, large-scale pathogen burden studies in children have identified ETEC in the guts of both symptomatic patients and controls. The factors that influence this balance are poorly understood, but it is postulated that the gut microbiome may play a role in either resistance or progression to disease. In this study, we profiled the microbiomes of children and adults from Bangladesh who were asymptomatically or symptomatically infected with ETEC. Symptomatic patients had significantly higher numbers of sequenced reads mapping to both E. coli and two ETEC toxins, suggesting higher bacterial burden. They were also significantly more likely to be coinfected with enteroaggregative E. coli (EAEC) and had higher proportions of other Gammaproteobacteria, including Klebsiella, Salmonella, and Haemophilus. Colonization with ETEC was also associated with increased prevalence of antimicrobial resistance (AMR) genes, most notably those of the ß-lactamase class. Taxonomic profiles were distinctly different between all groups in both species richness and composition, although the direction of these changes was different in adults and children. As seen previously, children with high E. coli burdens also had higher proportions of Streptococcus spp., while healthy children were more heavily colonized by Bifidobacterium spp. Our study provides insight into the microbiome changes that occur upon infection with ETEC in an endemic setting and provides rationale for future studies investigating how the microbiome may protect or predispose individuals to symptomatic infections with gastrointestinal pathogens. IMPORTANCE Enterotoxigenic Escherichia coli (ETEC) is an important cause of diarrhea in children in low- and middle-income countries. However, these bacteria are often identified in both patients and healthy controls. We do not yet understand why only some people get sick, but it has been suggested that the gut microbiome might play a role. In this study, we used metagenomic sequencing to profile the gut microbiomes of individuals in Bangladesh, with or without a symptomatic ETEC infection. In general, individuals with high levels of ETEC also harbored other pathogenic E. coli strains, higher proportions of Gammaproteobacteria such as Salmonella and Klebsiella, and a higher burden of antimicrobial resistance genes in their guts. Healthy children, in contrast, had higher levels of bifidobacteria. These data confirm that the composition of the gut microbiome is different between symptomatic and asymptomatic people and provides important preliminary information on the impact of the gut microbiome in intestinal infections.

Invasive weed optimization coupled biomass and product dynamics of tuning soybean husk towards lipolytic enzyme.

Waste to the product approach was proposed for tuning environ-threat soybean husk towards lipolytic enzyme by integrating the invasive weed optimization with biomass and product dynamics study. The invasive weed optimization constitutes based on the non-linear regression model results in a 47 % enhancement in lipolytic enzyme using the optimization parameters of 7% Sigma Final, 9% exponent; Smax of 5 with a population size of 35 and Max. generations of 99. The biomass dynamic study showcases the dynamic parameters of 0.0239 µmax, 8.17 XLimst and 0.852 RFin values. The product dynamic studies reveal the kinetic parameters of kst, kdiv, PFin, which seem to be equal to -0.0338, 0.0896 and 68.1, respectively. Overall, the present study put forth the zero-waste (soybean husk) to the product (lipolytic enzyme) approach by introducing the novel "Invasive Weed Optimization" coupled with "Biomass and product dynamics" to the bioprocessing field.

Phylogenetic and antimicrobial drug resistance analysis of Vibrio cholerae O1 isolates from Ghana.

We investigated the evolution, phylogeny and antimicrobial resistance of Vibrio cholerae O1 isolates (VCO1) from Ghana. Outbreak and environmental sources of VCO1 were characterized, whole-genome sequenced and compared to globally available seventh pandemic (7P) strains of V. cholerae at SNP resolution. Final analyses included 636 isolates. Novel Ghanaian isolates clustered into three distinct clades (clades 1, 2 and 3) in wave 3 of the 7P lineage. The closest relatives of our novel Ghanaian isolates were from Benin, Cameroon, Togo, Niger and Nigeria. All novel Ghanaian isolates were multi-drug resistant. Environmental isolates clustered into clade 2, despite being isolated years later, showing the possibility of persistence and re-emergence of older clades. A lag phase of several years from estimated introduction to reported cases suggests pathogen persistence in the absence of reported cholera cases. These results highlight the importance of deeper surveillance for understanding transmission routes between bordering countries and planning tailored vaccination campaigns in an effort to eradicate cholera.

Antimicrobial and dye degradation application of fungi-assisted silver nanoparticles and utilization of fungal retentate biomass for dye removal.

The present study utilized Aspergillus spp. for the synthesis of silver nanoparticles (AgNPs); the developed AgNPs were categorized using analytical techniques, that is, ultraviolet-visible (UV-vis) spectrophotometer, Zeta-potential, dynamic light scattering (DLS), and transmission electron microscopy (TEM). A sharp peak of 463 nm highlighted the synthesis of AgNPs; further Zeta-potential of -16 mV indicates stability of synthesized AgNPs. The TEM micrograph showed spherical and hexagonal shapes of synthesized AgNPs of 6-25 nm. The photocatalytic activity of fungal-mediated AgNPs was evaluated for degradation of reactive yellow dye in the concentration range of 20-100 mg L-1 . The results showed efficient degradation of dye using AgNPs in short span of time. For antibacterial activity, synthesized AgNPs, antibiotic, and AgNPs + antibiotic were tested. As per results, the zone of inhibition (ZOI) of AgNPs showed the values of 13 and 10 mm for Escherichia coli and Staphylococcus aureus, respectively. Further, the ZOI of penicillin highlighted the values of 18 and 17 mm for E. coli and S. aureus, respectively. When AgNPs and penicillin were used in combination, a clear synergistic effect was observed; the ZOI showed 0.49- and 0.36-fold increase in area against E. coli and S. aureus, respectively, in comparison with penicillin or AgNPs alone. Further, the leftover biomass (retentate biomass) was used to decolorize the reactive yellow dye at different initial concentration ranging from 20 to 100 mg L-1 . It was observed that 1 g L-1 retentate biomass (BR ) can effectively remove 82%-100% dye at 20 and 100 mg L-1 initial dye concentration. Results also indicated that with increase in initial reactive dye concentration from 20 to 100 mg L-1 , the decolorization capacity of retentate biomass (BR ) (at 0.2 g L-1 ) decreased from 79.2% to 32.3%. However, the use of AgNPs synthesized leftover fungal biomass can be a good option for up taking the additional dyes/contaminants, and also as leftover biomass can be utilized effectively, it can prove to be an excellent approach for environment safety. As the literature studies did not mentioned the further use of retentate biomass, the present study provides an excellent approach for further research on this aspect. PRACTITIONER POINTS: Synthesis of AgNPs from Aspergillus spp. and characterized with the help of a U.V-vis spectrophotometer, a zeta potential, DLS and TEM. The developed AgNPs were used for antibacterial and dye degradation activity. The left over (retentate) fungal biomass was used further for additional dye degradation activity.

Engineering aspects of immobilized lipases on esterification: A special emphasis of crowding, confinement and diffusion effects.

Cross-linked enzyme crystal (CLEC) and sol-gel entrapped pseudomonas sp. lipase were investigated for the esterification of lauric acid with ethanol by considering the effects of reaction conditions on reaction rate. The activation energy for the reaction was estimated to be 1097.58 J/mol and 181.75 J/mol for sol-gel and CLEC entrapped lipase respectively. CLEC lipase exhibited a marginal internal diffusion effect on reaction rate over sol-gel lipases and found to be interesting. The overall reaction mechanism was found to conform to the Ping Pong Bi Bi mechanism. The higher efficiency of sol-gel lipases over CLEC lipases in esterification reaction is mainly due to the combined effects of crowding, confinement and diffusional limitations.

Adoption of Electronic Health Records: A Roadmap for India.

OBJECTIVES: The objective of the study was to create a roadmap for the adoption of Electronic Health Record (EHR) in India based an analysis of the strategies of other countries and national scenarios of ICT use in India. METHODS: The strategies for adoption of EHR in other countries were analyzed to find the crucial steps taken. Apart from reports collected from stakeholders in the country, the study relied on the experience of the author in handling several e-health projects. RESULTS: It was found that there are four major areas where the countries considered have made substantial efforts: ICT infrastructure, Policy & regulations, Standards & interoperability, and Research, development & education. A set of crucial activities were identified in each area. Based on the analysis, a roadmap is suggested. It includes the creation of a secure health network; health information exchange; and the use of open-source software, a national health policy, privacy laws, an agency for health IT standards, R&D, human resource development, etc. CONCLUSIONS: Although some steps have been initiated, several new steps need to be taken up for the successful adoption of EHR. It requires a coordinated effort from all the stakeholders.

Study of structure-activity relationship of enantiomeric, protonated and deprotonated forms of warfarin via vibrational spectroscopy and DFT calculations.

The structure-activity relationship of the anticoagulant drug warfarin were studied by studying two enantiomeric forms (S-form and R-form) of warfarin and its protonated as well as deprotonated structures in aqueous media using density functional theory (DFT). Theoretically computed Raman and IR spectra of all the computed structures were compared and their specific vibrational spectroscopic signatures were discussed. The percentage contributions of individual normal modes of warfarin, which provides direct evidence of the different molecular activity due to change in relative atomic position of atoms in molecule, were investigated through potential energy distribution (PED). The optimized energy and molecular electrostatic potential (MEP) maps show that the S-form of the drug molecules warfarin is energetically more stable than R-form and provides higher docking opportunity for the molecular binding with the receptors in the bio-systems.

Modeling the weak hydrogen bonding of pyrrole and dichloromethane through Raman and DFT study.

Raman spectra of neat pyrrole (C(4)H(5)N) and its binary mixtures with dichloromethane (CH(2)Cl(2), DCM) with varying mole fractions of C(4)H(5)N from 0.1 to 0.9 were recorded in order to monitor the influence of molecular interaction on spectral features of selected vibrational bands of pyrrole in the region 600-1600 cm(-1). Only 1369 cm(-1) vibrational band of pyrrole shows a significant change in its peak position in going from neat pyrrole to the complexes. The 1369 cm(-1) band shows (∼6 cm(-1)) blue shift upon dilution and the corresponding linewidth shows the maximum shift at C = 0.5 mole fraction of pyrrole upon dilution which clearly indicates that the concentration fluctuation model plays major role. Quantum chemical calculation using density functional theory (DFT) and ab-initio (MP2 and HF) methods were performed employing high level basis set, 6-311++G(d,p) to obtain the ground state geometry of neat pyrrole and its complexes with DCM in gas phase. Basis set superimpose error (BSSE) correction was also introduced by using the counterpoise method. In order to account for the solvent effect on vibrational features and changes in optimized structural parameters of pyrrole, polarizable continuum model (PCM) (bulk solvations) and PCM (specific plus bulk solvations) calculations were performed. Two possible configurations of pyrrole + DCM complex have been predicted by B3LYP and HF methods, whereas the MP2 method gave only single configuration in which H atom of DCM is bonded to π ring of the pyrrole molecule. This affects significantly the ring vibrations of pyrrole molecule, which was also observed in our experimental results.