KNOWLEDGE ATTITUDE AND PRACTICE TOWARD COVID-19 DISEASE AMONG VARIOUS PROFESSIONALS: A CROSS-SECTIONAL ONLINE SURVEY

Objectives: This study aims to assess the knowledge, attitude, and practice associated with COVID-19 disease by undertaking a web-based online survey. Methods: A cross-sectional survey was conducted all over India during the initial phase of COVID-19 hit between March 19, 2020, and March 29, 2020. Anyone aged 18 years and above using social media were included in the study. The questionnaire had a section on sociodemography which had questions regarding age, gender, marital status, and occupation. This was followed by questions on knowledge, attitude, and practice regarding the COVID-19 pandemic. The knowledge scores were summarized as median with interquartile range as the data followed a non-normal distribution. Attitude and practice questions were summarized individually as proportions and its association with socio-demographic variables was established using Chi-square test. The statistical significance was set at p<0.05. Results: Occupational groups, marital status, and age group had significant difference between the groups with at least one of the practices with p<0.05. Conclusion: Since the study provides enhancing effect of knowledge toward protective behaviors, the awareness raising activities and health education through mass media and campaigns, this should urgently be conducted focusing on effectiveness of hand washing, cough etiquette, social distancing, and responsibility to inform suspected cases to local health authority to prevent COVID-19

TSH Receptor Gene and Autoimmune Thyroid Diseases

The primary regulators of thyroid activity are the thyroid-stimulating hormone (TSH) and its receptor (TSH-R). Studies have shown that genetic variants in the TSHR gene can increase susceptibility to autoimmune thyroid diseases (AITD). The TSHR gene is located on chromosome 14q31 and encodes a membrane-bound receptor that interacts with TSH to regulate thyroid hormone synthesis and secretion. AITD including Graves' disease (GD) and Hashimoto's thyroiditis (HT), are the most common thyroid disorders, affecting millions of people worldwide. In AITD, autoantibodies can bind to and activate the TSHR, leading to increased thyroid hormone production and secretion in GD, or thyroid destruction and hypothyroidism in HT. In addition to its role in thyroid hormone synthesis and secretion, some studies also revealed that the TSHR has also been implicated in a variety of other physiological processes, including bone metabolism, reproduction, and immune regulation. Genetic variation in the TSHR region may affect the expression, post-translational processing, and/or protein structure, which in turn may cause or worsen the autoimmune response. The TSHR gene and its products are widely used in diagnostic testing for AITD. Understanding the molecular mechanisms underlying the interaction between the TSHR and autoantibodies is critical for developing new diagnostic and therapeutic strategies for AITD

Multifactorial Aspects Influencing Non-Alcoholic Fatty Liver Disease (Nafld)

Nonalcoholic fatty liver disease (NAFLD) is a growing public health concern, with a prevalence of up to 25% worldwide. While once considered a benign condition, NAFLD is now recognized as a major cause of chronic liver disease, liver failure, and hepatocellular carcinoma. The pathogenesis of NAFLD is multifactorial and involves a complex interplay between genetic, environmental, and metabolic factors. In this review, we provide an overview of the multifactorial aspects of NAFLD, including genetic predisposition, insulin resistance, dyslipidemia, gut microbiota, dietary factors, and physical inactivity. We also discuss the role of inflammation, oxidative stress, and hepatic steatosis in the progression of NAFLD to nonalcoholic steatohepatitis (NASH), cirrhosis, and hepatocellular carcinoma. Finally, we review the current and emerging therapies for NAFLD and NASH, including lifestyle modifications, pharmacological interventions, and surgical approaches. The multifactorial nature of NAFLD requires a comprehensive approach to diagnosis, treatment, and prevention, with a focus on addressing the underlying metabolic and environmental factors that contribute to its development and progression

Genetic Aspects of Implantation Failure

Implantation failure refers to the inability of a fertilized egg, or embryo, to successfully implant itself in the endometrial lining of the uterus, leading to pregnancy loss. The repeated failure of good quality embryo implantation is referred to as recurrent implantation failure (RIF). This can occur for a variety of reasons, including chromosomal abnormalities in the embryo, problems with the endometrium, or issues with the immune system. Factors such as advanced maternal age, obesity, smoking, and certain medical conditions can also increase the risk of implantation failure. While treatment such as in vitro fertilization (IVF) can help to improve the chances of successful implantation, there is currently no definite way to prevent or treat implantation failure.  Patients and healthcare professionals have substantial diagnostic and treatment hurdles as a result of many etiological factors and lack of knowledge about RIF. A number of studies have indicated a correlation between irregular hormone levels, disruptions in angiogenic and immunomodulatory factors, specific genetic polymorphisms, and the prevalence of RIF. Nonetheless, the precise and intricate underlying pathophysiology of RIF remains elusive.  However, many studies are ongoing in this field to understand the underlying causes and to find new ways to help couples achieve pregnancy. This review article extensively explores diverse molecular and genetic facets aimed at enhancing the diagnosis and management of implantation failure

Novel Role of Phosphorylation-Dependent Interaction between FtsZ and FipA in Mycobacterial Cell Division

The bacterial divisome is a multiprotein complex. Specific protein-protein interactions specify whether cell division occurs optimally, or whether division is arrested. Little is known about these protein-protein interactions and their regulation in mycobacteria. We have investigated the interrelationship between the products of the Mycobacterium tuberculosis gene cluster Rv0014c-Rv0019c, namely PknA (encoded by Rv0014c) and FtsZ-interacting protein A, FipA (encoded by Rv0019c) and the products of the division cell wall (dcw) cluster, namely FtsZ and FtsQ. M. smegmatis strains depleted in components of the two gene clusters have been complemented with orthologs of the respective genes of M. tuberculosis. Here we identify FipA as an interacting partner of FtsZ and FtsQ and establish that PknA-dependent phosphorylation of FipA on T77 and FtsZ on T343 is required for cell division under oxidative stress. A fipA knockout strain of M. smegmatis is less capable of withstanding oxidative stress than the wild type and showed elongation of cells due to a defect in septum formation. Localization of FtsQ, FtsZ and FipA at mid-cell was also compromised. Growth and survival defects under oxidative stress could be functionally complemented by fipA of M. tuberculosis but not its T77A mutant. Merodiploid strains of M. smegmatis expressing the FtsZ(T343A) showed inhibition of FtsZ-FipA interaction and Z ring formation under oxidative stress. Knockdown of FipA led to elongation of M. tuberculosis cells grown in macrophages and reduced intramacrophage growth. These data reveal a novel role of phosphorylation-dependent protein-protein interactions involving FipA, in the sustenance of mycobacterial cell division under oxidative stress

An Oligopeptide Transporter of Mycobacterium tuberculosis Regulates Cytokine Release and Apoptosis of Infected Macrophages

Background: The Mycobacterium tuberculosis genome encodes two peptide transporters encoded by Rv3665c-Rv3662c and Rv1280c-Rv1283c. Both belong to the family of ABC transporters containing two nucleotide-binding subunits, two integral membrane proteins and one substrate-binding polypeptide. However, little is known about their functions in M. tuberculosis. Here we report functional characterization of the Rv1280c-Rv1283c-encoded transporter and its substrate-binding polypeptide OppA(MTB). Methodology/Principal Findings: OppA(MTB) was capable of binding the tripeptide glutathione and the nonapeptide bradykinin, indicative of a somewhat broad substrate specificity. Amino acid residues G109, N110, N230, D494 and F496, situated at the interface between domains I and III of OppA, were required for optimal peptide binding. Complementaton of an oppA knockout mutant of M. smegmatis with OppA(MTB) confirmed the role of this transporter in importing glutathione and the importance of the aforesaid amino acid residues in peptide transport. Interestingly, this transporter regulated the ability of M. tuberculosis to lower glutathione levels in infected compared to uninfected macrophages. This ability was partly offset by inactivation of oppD. Concomitantly, inactivation of oppD was associated with lowered levels of methyl glyoxal in infected macrophages and reduced apoptosis-inducing ability of the mutant. The ability to induce the production of the cytokines IL-1 beta, IL-6 and TNF-alpha was also compromised after inactivation of oppD. Conclusions: Taken together, these studies uncover the novel observations that this peptide transporter modulates the innate immune response of macrophages infected with M. tuberculosis

Understanding the Role of PknJ in Mycobacterium tuberculosis: Biochemical Characterization and Identification of Novel Substrate Pyruvate Kinase A

Reversible protein phosphorylation is a prevalent signaling mechanism which modulates cellular metabolism in response to changing environmental conditions. In this study, we focus on previously uncharacterized Mycobacterium tuberculosis Ser/Thr protein kinase (STPK) PknJ, a putative transmembrane protein. PknJ is shown to possess autophosphorylation activity and is also found to be capable of carrying out phosphorylation on the artificial substrate myelin basic protein (MyBP). Previous studies have shown that the autophosphorylation activity of M. tuberculosis STPKs is dependent on the conserved residues in the activation loop. However, our results show that apart from the conventional conserved residues, additional residues in the activation loop may also play a crucial role in kinase activation. Further characterization of PknJ reveals that the kinase utilizes unusual ions (Ni2+, Co2+) as cofactors, thus hinting at a novel mechanism for PknJ activation. Additionally, as shown for other STPKs, we observe that PknJ possesses the capability to dimerize. In order to elucidate the signal transduction cascade emanating from PknJ, the M. tuberculosis membrane-associated protein fraction is treated with the active kinase and glycolytic enzyme Pyruvate kinase A (mtPykA) is identified as one of the potential substrates of PknJ. The phospholabel is found to be localized on serine and threonine residue(s), with Ser37 identified as one of the sites of phosphorylation. Since Pyk is known to catalyze the last step of glycolysis, our study shows that the fundamental pathways such as glycolysis can also be governed by STPK-mediated signaling

Bacterial Transmembrane Proteins that Lack N-Terminal Signal Sequences

Tail-anchored membrane proteins (TAMPs), a class of proteins characterized by their lack of N-terminal signal sequence and Sec-independent membrane targeting, play critical roles in apoptosis, vesicle trafficking and other vital processes in eukaryotic organisms. Until recently, this class of membrane proteins has been unknown in bacteria. Here we present the results of bioinformatic analysis revealing proteins that are superficially similar to eukaryotic TAMPs in the bacterium Streptomyces coelicolor. We demonstrate that at least four of these proteins are bona fide membrane-spanning proteins capable of targeting to the membrane in the absence of their N-terminus and the C-terminal membrane-spanning domain is sufficient for membrane targeting. Several of these proteins, including a serine/threonine kinase and the SecE component of the Sec translocon, are widely conserved in bacteria

Appropriate DevR (DosR)-Mediated Signaling Determines Transcriptional Response, Hypoxic Viability and Virulence of Mycobacterium tuberculosis

Background: The DevR(DosR) regulon is implicated in hypoxic adaptation and virulence of Mycobacterium tuberculosis. The present study was designed to decipher the impact of perturbation in DevR-mediated signaling on these properties. Methodology/Principal Findings: M. tb complemented (Comp) strains expressing different levels of DevR were constructed in Mut1 * background (expressing DevR N-terminal domain in fusion with AphI (DevRN-Kan) and in Mut2DdevR background (deletion mutant). They were compared for their hypoxia adaptation and virulence properties. Diverse phenotypes were noted; basal level expression (,5.362.3 mM) when induced to levels equivalent to WT levels (,25.869.3 mM) was associated with robust DevR regulon induction and hypoxic adaptation (Comp 9 * and 10*), whereas low-level expression (detectable at transcript level) as in Comp 11 * and Comp15 was associated with an adaptation defect. Intermediate-level expression (,3.361.2 mM) partially restored hypoxic adaptation functions in Comp2, but not in Comp1 * bacteria that coexpressed DevRN-Kan. Comp * strains in Mut1 * background also exhibited diverse virulence phenotypes; high/very low-level DevR expression was associated with virulence whereas intermediate-level expression was associated with low virulence. Transcription profiling and gene expression analysis revealed up-regulation of the phosphate starvation response (PSR) in Mut1 * and Comp11 * bacteria, but not in WT/Mut2DdevR/other Comp strains, indicating a plasticity in expression pathways that is determined by the magnitude of signaling perturbation through DevRN-Kan