Mental health morbidities in Kerala, India: Insights from National Mental Health Survey, 2015-2016.

Background: The National Mental Health Survey was borne out of the felt need for a comprehensive epidemiological survey on mental health to understand the magnitude of psychiatric morbidities in India to aid in mental health policymaking, service planning, and delivery. Kerala was one of the 12 surveyed states, representing southern India. Aims: To estimate the prevalence and pattern of various mental illnesses and substance use disorders in a representative sample from Kerala state. Settings and Design: A household survey using a multi-stage, stratified, random cluster sampling technique, with selection based on probability proportionate to size at each stage. Materials and Methods: The community-based survey was carried out by trained field staff on individuals from systematically selected households from three randomly selected districts of Kerala. The instruments used in the survey included M.I.N.I adult version 6.0, a modified version of the Fagerström Nicotine Dependence Scale and questionnaires to screen for epilepsy, intellectual disability, and autism spectrum disorders. Results: A total of 2479 respondents aged >18 years were interviewed. The lifetime and current prevalence of mental morbidity (excluding tobacco use disorders) was 14.14% and 11.36%, respectively. Neurotic/stress-related disorders and depressive disorders were 5.43% and 2.49%, respectively, while severe mental disorders were prevalent in 0.44% of the sample. The prevalence of high risk for suicide was 2.23%. Conclusions: The survey revealed high rates of common mental illnesses and suicide risk in the state when compared to national estimates.

Bile Salts Promote ToxR Regulon Activation during Growth under Virulence-Inducing Conditions.

Cholera is an epidemic disease caused by the Gram-negative bacterium Vibrio cholerae. V. cholerae is found in aquatic ecosystems and infects people through the consumption of V. cholerae-contaminated food or water. Following ingestion, V. cholerae responds to host cues to activate the expression of critical virulence genes that are under the control of a hierarchical regulatory system called the ToxR regulon. The ToxR regulon is tightly regulated and is expressed in vitro only under special growth conditions referred to as AKI conditions. AKI conditions have been instrumental in elucidating V. cholerae virulence regulation, but the chemical cues within AKI medium that activate virulence gene expression are unknown. In this study, we fractionated AKI medium on a reverse-phase chromatography column (RPCC) and showed that the virulence-activating molecules were retained on the RPCC column and recovered in the eluate. Liquid chromatography-high-resolution mass spectrometry (LC-HRMS) analysis of the eluate revealed the presence of a known ToxR regulon activator, taurocholate, and other bile salts. The RPCC eluate activated the ToxR regulon when added to noninducing medium and promoted TcpP dimerization in a two-hybrid system, consistent with taurocholate being responsible for the virulence-inducing activity of AKI medium. Additional experiments using purified bile salts showed that the ToxR regulon was preferentially activated in response to primary bile acids. The results of this study shed light on the chemical cues involved in V. cholerae virulence activation and suggested that V. cholerae virulence genes are modulated in response to regionally specific bile acid species in the intestine.

Vibrio cholerae TolC Is Required for Expression of the ToxR Regulon.

Vibrio cholerae is a Gram-negative bacterium that causes the enteric disease cholera. V. cholerae colonization of the human intestine is dependent on the expression of both virulence genes and environmental adaptation genes involved in antimicrobial resistance. The expression of virulence genes, including the genes encoding the main virulence factors cholera toxin (CT) and the toxin-coregulated pilus (TCP), are coordinately regulated by the ToxR regulon. Tripartite transport systems belonging to the ATP binding cassette, major facilitator, and resistance-nodulation-division families are critical for V. cholerae pathogenesis. Transport systems belonging to these families contribute to myriad phenotypes, including protein secretion, antimicrobial resistance, and virulence. TolC plays a central role in bacterial physiology by functioning as the outer membrane pore protein for tripartite transport systems. Consistent with this, V. cholerae tolC was previously found to be required for MARTX toxin secretion and antimicrobial resistance. Here, we investigated the contribution of TolC to V. cholerae virulence. We documented that tolC was required for CT and TCP production in O1 El Tor V. cholerae. This phenotype was linked to repression of the critical ToxR regulon transcription factor aphA. Decreased aphA transcription correlated with increased expression of the LysR-family transcription factor leuO. Deletion of leuO restored aphA expression, and CT and TCP production, in a tolC mutant. The collective results document that tolC is required for ToxR regulon expression and further suggest that tolC participates in an efflux-dependent feedback circuit to regulate virulence gene expression.

ToxR Mediates the Antivirulence Activity of Phenyl-Arginine-ß-Naphthylamide To Attenuate Vibrio cholerae Virulence.

Multidrug efflux systems belonging to the resistance-nodulation-cell division (RND) family are ubiquitous in Gram-negative bacteria and are critical for antimicrobial resistance. This realization has led to efforts to develop efflux pump inhibitors (EPI) for use as adjuvants for antibiotic treatment of resistant organisms. However, the functions of RND transporters extend beyond antimicrobial resistance to include physiological functions that are critical for pathogenesis, suggesting that EPIs could also be used as antivirulence therapeutics. This was documented in the enteric pathogen Vibrio cholerae, in which EPIs were shown to attenuate the production of the critical virulence factors cholera toxin (CT) and the toxin-coregulated pilus (TCP). In this study, we investigated the antivirulence mechanism of action of the EPI phenyl-arginine-ß-naphthylamide (PAßN) on V. cholerae. Using bioassays, we documented that PAßN inhibited virulence factor production in three epidemic V. cholerae isolates. Transcriptional reporter studies and mutant analysis indicated that PAßN initiated a ToxR-dependent regulatory circuit to activate leuO expression and that LeuO repressed the expression of the critical virulence activator aphA to attenuate CT and TCP production. The antivirulence activity of PAßN was found to be dependent on the ToxR periplasmic sensing domain (PPD), suggesting that a feedback mechanism was involved in its activity. Collectively, the data indicated that PAßN inhibited V. cholerae virulence factor production by activating a ToxR-dependent metabolic feedback mechanism to repress the expression of the ToxR virulence regulon. This suggests that efflux pump inhibitors could be used as antivirulence therapeutics for the treatment of cholera and perhaps that of other Gram-negative pathogens.

NexusLIMS: A Laboratory Information Management System for Shared-Use Electron Microscopy Facilities.

This work introduces NexusLIMS, an electron microscopy laboratory information management system designed and implemented by the Office of Data and Informatics and the Materials Science and Engineering Division at NIST for a multi-user electron microscopy co-op facility. NexusLIMS comprises network infrastructure, shared information technology resources, a custom software package to harvest and extract experimental information and construct experimental metadata records, and an intuitive web-based user-facing interface for searching, browsing, and examining research data. These metadata records conform to the Nexus Experiment schema, which is introduced in this work. The NexusLIMS suite of tools requires minimal input and adjustments to user behavior, instead relying on existing organizational procedures and the collection of information from a multitude of sources to construct a complete picture and record of a research experiment. The underlying infrastructure and design considerations for a multi-user data management system are discussed. The core codebase for NexusLIMS is made publicly available alongside this work, and its modular design encourages the adaptation of the presented methods at other research organizations.

Disability with Persistent Pain Following an Epidemic of Chikungunya in Rural South India.

OBJECTIVE: We investigated the effects of chronic rheumatic and musculoskeletal symptoms on the functional status of people affected by the chikungunya (CKG) epidemic in the Calicut District, Kerala, South India in 2009. METHODS: A cross-sectional house-to-house survey was conducted 18 months after the CKG epidemic to assess functional status of individuals with post-epidemic persistent pain. All respondents over age 15 years with persistent pain fitting the epidemiological case definition were included. Participants' functional status was assessed using the Health Assessment Questionnaire-Disability Index (HAQ-DI). Factors affecting severity of HAQ-DI were analyzed by ordinal regression. RESULTS: Of 3869 subjects interviewed, 1195 (34.3%) had a positive history of CHIKV virus infection (epidemiological or confirmed); 36.28% (624/1720) of CKG-affected individuals had persistent pain 18 months post epidemic. Mean age of those affected was 48.22 ± 15.6 years; 23.2% had no disability, while 16.2% had moderate to severe disability on the HAQ-DI. Significant factors affecting severity of disability on HAQ-DI included previous rheumatic musculoskeletal disease (OR 2.27), joint and soft-tissue involvement (OR 3.74), only joint involvement (OR 2.14), female sex (OR 1.44), diet (OR 4.73), and history of joint swelling (OR 1.72). CONCLUSION: Persistence of pain noted in post-CKG disease resulted in significantly deteriorated functional status of those affected.

Construction of a tetracycline inducible expression vector and characterization of its use in Vibrio cholerae.

We report the construction of a tetracycline inducible expression vector that allows regulated gene expression in the enteric pathogen Vibrio cholerae. The expression vector, named pXB300, contains the tetracycline regulatory elements from Tn10, a multiple cloning site downstream of the tetA promoter and operator sequences, a ColE1 origin of replication, a ß-lactamase resistance gene for positive selection, and the hok/sok addiction system for selection in the absence of antibiotic. The function of the tetracycline expression system was demonstrated by cloning lacZ under control of the tetA promoter and quantifying ß-galactosidase expression in Escherichia coli and V. cholerae. The utility for pXB300 was documented by complementation of V. cholerae virulence mutants during growth under virulence inducing conditions. The results showed that pXB300 allowed high-level expression of recombinant genes with linear induction in response to the exogenous concentration of the inducer anhydrotetracycline. We further show that pXB300 was reliably maintained in V. cholerae during growth in the absence of antibiotic selection.

Prevalence and factors related to rheumatic musculoskeletal disorders in rural south India: WHO-ILAR-COPCORD-BJD India Calicut study.

AIMS: To assess the prevalence and factors related to rheumatic musculoskeletal disorders (RMSD) in a rural population of south India. METHODS: The cross-sectional study included all individuals, 15 years and above, in a rural unit of Calicut District in North Kerala. Data were collected using the validated World Health Organization - International League of Associations for Rheumatology - Community Oriented Program for the Control of Rheumatic Diseases - Bhigwan model questionnaire by trained volunteers. In Phase 1 details of demographic characteristics, major co-morbidities and perceived musculoskeletal aches and pains were elicited. Phases 2 and 3 further evaluated and diagnosed the subjects. Predictors for RMSD were assessed using binary logistic regression analysis. RESULTS: There were 4999 individuals in the study. The prevalence of RMSD was 24.9% (95% CI 23.73; 26.12%). Females constituted 50.7% of the population; 5.1% of the respondents were illiterate; 80.9% belonged to low-income groups. Diabetes mellitus and hypertension affected 4.1% and 5.4% of the subjects respectively. The predictors for RMSD in the population were female sex, age, illiteracy, married status, low-income group, vegetarian diet, current alcohol consumption, current tobacco use, history of injury or accidents, diabetes and hypertension. Symptom-related ill-defined rheumatism (10.39%) followed by osteoarthritis (3.85%) were the most prevalent in the Phase 3 rheumatological evaluation. CONCLUSION: There is an urgent need to introduce lifestyle modifications in high-risk groups and start rehabilitation for those affected. Community rheumatology in primary health care settings in rural areas needs to be strengthened by introducing national programs addressing RMSD at the grassroots level.