Determinants Outside the DevR C-Terminal Domain Are Essential for Cooperativity and Robust Activation of Dormancy Genes in Mycobacterium tuberculosis

Background: DevR (also called as DosR) is a two-domain response regulator of the NarL subfamily that controls dormancy adaptation of Mycobacterium tuberculosis (M. tb). In response to inducing signals such as hypoxia and ascorbic acid, the N-terminal receiver domain of DevR (DevRN) is phosphorylated at Asp54. This results in DevR binding to DNA via its C-terminal domain (DevRC) and subsequent induction of the DevR regulon. The mechanism of phosphorylation-mediated activation is not known. The present study was designed to understand the role of the N- and C-terminal domains of DevR in DevR regulon genes activation. Methodology/Principal Findings: Towards deciphering the activation mechanism of DevR, we compared the DNA binding properties of DevRC and DevR and correlated the findings with their ability to activate gene expression. We show that isolated DevRC can interact with DNA, but only with the high affinity site of a representative target promoter. Therefore, one role of DevR N is to mask the intrinsic DNA binding function of DevR C. However, unlike phosphorylated DevR, isolated DevR C does not interact with the adjacent low affinity binding site suggesting that a second role of DevRN is in cooperative binding to the secondary site. Transcriptional analysis shows that consistent with unmasking of its DNA binding property, DevRC supports the aerobic induction, albeit feebly, of DevR regulon genes but is unable to sustain gene activation during hypoxia

Use, abuse, and associated impacts of alcohol on health and crimes in Nepal

Alcohol is a socially acceptable and widely available drink in most countries. Its excessive consumption is linked to various health issues, increase in crime rate and even loss of life. This puts heavy strain in low-income countries like Nepal where full medical facilities are still out of reach for most of the population. Therefore, in this article, we have provided an overview of (i) the effect of alcohol on public health; (ii) trend data on alcohol seizures; (iii) alcohol (including methanol) positive forensic cases; and (iv) other alcohol associated crimes. Our analysis is drawn from a range of data types and sources, triangulating the collected data with alcohol specific academic and grey literature, a survey with students and insights from stakeholder engagements in Nepal. We have shown that alcohol has been associated with a range of health-related issues and crime types in Nepal. For example, alcohol was the most mentioned compound in the student survey (50.7%; n=418) followed by other drugs in Drugs Facilitated Sexual Assault cases. We have also discussed issues with adulteration, surrogates and sub-standard alcohol, highlighting the need for strict monitoring, regulations and extra vigilance about quality control of alcohol in circulation. This should be supported by public awareness campaigns on the use, abuse and impacts of alcohol

Development and testing the feasibility of a sports-based mental health promotion intervention in Nepal: a protocol for a pilot cluster-randomised controlled trial

Background: Mental wellbeing encompasses life satisfaction, social connectedness, agency and resilience. In adolescence, mental wellbeing reduces sexual health risk behaviours, substance use and violence; improves educational outcomes; and protects mental health in adulthood. Mental health promotion seeks to improve mental wellbeing and can include activities to engage participants in sport. However, few high-quality trials of mental health promotion interventions have been conducted with adolescents, especially in low- and middle-income countries. We sought to address this gap by testing SMART (Sports-based Mental heAlth pRomotion for adolescenTs) in a pilot cluster-randomised controlled trial (cRCT) in Bardiya, Nepal. // Methods: The objectives of the trial are to assess the acceptability and feasibility of SMART, test trial procedures, explore outcome distributions in intervention and control clusters and calculate the total annual cost of the intervention and unit cost per adolescent. The trial design is a parallel-group, two-arm superiority pilot cRCT with a 1:1 allocation ratio and two cross-sectional census surveys with adolescents aged 12–19, one pre-intervention (baseline) and one post-intervention (endline). The study area is four communities of approximately 1000 population (166 adolescents per community). Each community represents one cluster. SMART comprises twice weekly football, martial arts and dance coaching, open to all adolescents in the community, led by local sports coaches who have received psychosocial training. Sports melas (festivals) and theatre performances will raise community awareness about SMART, mental health and the benefits of sport. Adolescents in control clusters will participate in sport as usual. In baseline and endline surveys, we will measure mental wellbeing, self-esteem, self-efficacy, emotion regulation, social support, depression, anxiety and functional impairment. Using observation checklists, unstructured observation and attendance registers from coaching sessions, and minutes of meetings between coaches and supervisors, we will assess intervention fidelity, exposure and reach. In focus group discussions and interviews with coaches, teachers, caregivers and adolescents, we will explore intervention acceptability and mechanisms of change. Intervention costs will be captured from monthly project accounts, timesheets and discussions with staff members. // Discussion: Findings will identify elements of the intervention and trial procedures requiring revision prior to a full cRCT to evaluate the effectiveness of SMART

Comprehensive insights into Mycobacterium tuberculosis DevR (DosR) regulon activation switch

DevR regulon function is believed to be crucial for the survival of Mycobacterium tuberculosis during dormancy. In this study, we undertook a comprehensive analysis of the DevR regulon. All the regulon promoters were assigned to four classes based on the number of DevR binding sites (Dev boxes). A minimum of two boxes are essential for complete interaction and their tandem arrangement is an architectural hallmark at all promoters. Initial interaction of DevR with the conserved box is essential for its cooperative binding to adjacent sites bearing low to very poor sequence conservation and is the universal mechanism underlying DevR-mediated transcriptional induction. The functional importance of tandem arrangement was established by analyzing promoter variants harboring Dev boxes with altered spacing. Conserved sequence logos were generated from 47 binding sequences which included 24 newly discovered Dev boxes. In each half site of an 18-bp binding motif, G5 and C7 are essential for DevR binding. Finally, we show that DevR regulon induction occurs in a temporal manner and genes that are induced early are also usually powerfully induced. The information theory-based approach along with binding and temporal expression studies provide us with comprehensive insights into the complex pattern of DevR regulon activation

GDF15 promotes weight loss by enhancing energy expenditure in muscle

Funding Information: We thank R. Seeley for sharing GFRAL-null mice; B. Lowell for sharing β-less mice; and J. Wu for shipping β-less mice to us. G.R.S. was supported by a Diabetes Canada Investigator Award (DI-5-17-5302-GS), a Canadian Institutes of Health Research Foundation Grant (201709FDN-CEBA-116200), a Tier 1 Canada Research Chair in Metabolic Diseases and a J. Bruce Duncan Endowed Chair in Metabolic Diseases; D.W. by Fellowship Grants from the McMaster Institute for Research on Aging (MIRA) at McMaster University; S.R. by a postdoctoral fellowship supported by MITACS and Novo Nordisk; L.K.T. by a CIHR Post-Doctoral Fellowship Award and Michael DeGroote Fellowship Award in Basic Biomedical Science; E.M.D. by a Vanier Canada Graduate Scholarship; G.P.H. by the Natural Sciences and Engineering Research Council of Canada (NSERC: 400362); G.J.D. and S.M.F. by NSERC-CGSM scholarships; L.D. by the Fonds de Recherche du Québec-Santé doctoral training award; D.P.B. by the GSK Chair in Diabetes of Université de Sherbrooke and a FRQS J1 salary award. The Genotype-Tissue Expression (GTEx) Project was supported by the Common Fund of the Office of the Director of the National Institutes of Health, and by the NCI, NHGRI, NHLBI, NIDA, NIMH and NINDS. Funding Information: S.B.J. and R.E.K. are employees of Novo Nordisk, a pharmaceutical company producing and selling medicine for the treatment of diabetes and obesity. G.R.S. is a co-founder and shareholder of Espervita Therapeutics. McMaster University has received funding from Espervita Therapeutics, Esperion Therapeutics, Poxel Pharmaceuticals and Nestle for research conducted in the laboratory of G.R.S. S.R. is supported by a MITACS postdoctoral fellowship sponsored by Novo Nordisk. H.C.G. holds the McMaster-Sanofi Population Health Institute Chair in Diabetes Research and Care. G.R.S., G.P. and H.C.G. are inventors listed on a patent for identifying GDF15 as a biomarker for metformin. G.R.S. has received consulting/speaking fees from Astra Zeneca, Eli Lilly, Esperion Therapeutics, Merck, Poxel Pharmaceuticals and Cambrian Biosciences. The other authors declare no competing interests. Publisher Copyright: © 2023, The Author(s).Peer reviewedPublisher PD

Case report: An interesting case of large colloid goiter with dyspnea

Benign nodular goitre is one of the most common endocrine disorder, especially in countries like Nepal where iodine deficiency is endemic. Case History: A 59 years old female presented with large thyroid swelling. Investigations showed it to be colloid goiter compressing the airway. Left hemithyroidectomy was done. Post operative period was uneventful.Discussion: Management of thyroid nodules depends upon assessment of thyroid function, fine needle aspiration cytology and imaging. Surgery is the common method of treatment.   </p

Ocular morbidity among children at a tertiary eye care hospital in Kolkata, West Bengal

Eye diseases in childhood are important causes of medical consultation and it affects learning ability, adjustment in school and personality. To determine the pattern of ocular morbidity a cross-sectional observational study was conducted among 714 children, attending Ophthalmology department at a tertiary eye care center in Kolkata, West Bengal. All the children less than 15 years of age, attending in Unit II outpatient department were selected by complete enumeration method for duration of one year (January-December 2010). Distribution of association was analyzed by Chi-square test and difference between two proportions was calculated by z test for proportions. The common ocular morbidity were refractive errors (23.67%) followed by allergic conjunctivitis (17.23%), infection of the eye and adnexa (15.13%), ocular trauma (12.74%), and congenital eye diseases (13.59%). Majority of ocular morbidity is treatable and need early attention through eye screening cum intervention program beginning right from the childhood

Structure-Based Design of DevR Inhibitor Active against Nonreplicating Mycobacterium tuberculosis

Antitubercular treatment is directed against actively replicating organisms. There is an urgent need to develop drugs targeting persistent subpopulations of Mycobacterium tuberculosis. The DevR response regulator is believed to play a key role in bacterial dormancy adaptation during hypoxia. We developed a homology-based model of DevR and used it for the rational design of inhibitors. A phenylcoumarin derivative (compound 10) identified by in silico pharmacophore-based screening of 2.5 million compounds employing protocols with some novel features including a water-based pharmacophore query, was characterized further. Compound 10 inhibited DevR binding to target DNA, down-regulated dormancy genes transcription, and drastically reduced survival of hypoxic but not nutrient-starved dormant bacteria or actively growing organ ` isms. Our findings suggest that compound 10 ``locks'' DevR in an inactive conformation that is unable to bind cognate DNA and induce the dormancy regulon. These results provide proof-of-concept for DevR as a novel target to develop molecules with sterilizing activity against tubercle bacilli

DevR regulon expression in DevR_C-expressing cultures declines during hypoxia.

<p>(<b>A</b>) <i>M. tb</i> lysates (15 µg protein) were immunoblotted using rabbit anti-HspX or anti-SigA polyclonal sera and the blots were analyzed using Quantity One software (Bio-Rad, USA). The normalised intensities of the HspX-derived signals (with respect to those of SigA) are denoted as Arbitrary Signal Intensities (ASI) with respect to those obtained in 5 days hypoxic WT cultures (H5). ‘Aer’, aerobic; H1 H3 and H5 refer to 1, 3 and 5 days hypoxic cultures, respectively; ND, not detected. (<b>B</b>) Relative Quantity (RQ) of <i>devR_C</i> transcripts in different Comp strains determined by real time RT-PCR analysis. (<b>C</b>) Real time RT-PCR analysis of DevR regulon transcripts. Fold change in the relative quantity of transcripts under ‘hypoxic’ vs. ‘aerobic’ conditions (<b>fold decrease</b> in Comp5 and <b>fold increase</b> in Comp13) is shown.</p