Cyclic AMP binding to a universal stress protein in Mycobacterium tuberculosis is essential for viability.

Mycobacterial genomes encode multiple adenylyl cyclases and cAMP effector proteins, underscoring the diverse ways these bacteria utilize cAMP. We identified universal stress proteins, Rv1636 and MSMEG_3811 in Mycobacterium tuberculosis and Mycobacterium smegmatis, respectively, as abundantly expressed, novel cAMP-binding proteins. Rv1636 is secreted via the SecA2 secretion system in M. tuberculosis but is not directly responsible for the efflux of cAMP from the cell. In slow-growing mycobacteria, intrabacterial concentrations of Rv1636 were equivalent to the concentrations of cAMP present in the cell. In contrast, levels of intrabacterial MSMEG_3811 in M. smegmatis were lower than that of cAMP and therefore, overexpression of Rv1636 increased levels of "bound" cAMP. While msmeg_3811 could be readily deleted from the genome of M. smegmatis, we found that the rv1636 gene is essential for the viability of M. tuberculosis and is dependent on the cAMP-binding ability of Rv1636. Therefore, Rv1636 may function to regulate cAMP signaling by direct sequestration of the second messenger. This is the first evidence of a "sponge" for any second messenger in bacterial signaling that would allow mycobacterial cells to regulate the available intrabacterial "free" pool of cAMP.

Prevalence and predictors of low bone mineral density among adults aged 50 years and above in Chandigarh, India.

Background: Osteoporosis, defined as skeletal disorder characterized by reduced bone density and strength, is prevalent in both developing and developed countries. Dual energy X-ray absorptiometry (DEXA) is an effective tool to diagnose low bone mineral density (BMD). Objective: This study objective was to know the prevalence and predictors of low BMD in a community-based screening program among the adult population using DEXA scan. Methods: In this community-based study conducted between January and June 2016, a population-based screening program was offered to both males and females of age 50 years and above. BMD was measured to derive T-scores and classify as normal, osteopenic, and osteoporotic individuals. Results: A total of 455 subjects were enrolled where 62.4% proportion was women; 42.6% subjects were in the age group of 50 to 59 years. The prevalence of osteoporosis and osteopenia was 20.9% and 48.1%, respectively. Hypertension (23.7%) and diabetes (8.4%) were common comorbidities. Age and gender (female) were independent predictors of low BMD. Conclusion: Osteoporosis/osteopenia is present in both genders with increasing trend with age advancement. Institution of timely screening will prevent the morbidity associated with fractures due to low BMD.

Coverage vs Utilization of integrated child services scheme (ICDS): A community based study in urban block of Patiala, Punjab (India).

Introduction: Integrated Child Development Services (ICDS) scheme provides a wide range of health, nutrition and education services to children, women and adolescent girls. Maternal satisfaction usually influences service uptake and utilization. Therefore, in addition to coverage, the quality of services provided and end-user satisfaction need to be assessed. Material and Methods: A cross-sectional community-level study was conducted in the urban block of the Patiala district in Punjab. A total of 54 AWCs were selected from a sample frame of 222 by applying a systematic random sampling technique. From each selected AWC, 20 mothers of registered beneficiaries (0-6 years) were randomly selected for interview. The data collected was entered and analysed using SPSS version 22. Results: The present study revealed that most of the Anganwadis (87.19%) were regularly open for the provision of various ICDS services. However, only 48.02% of children received supplementary nutrition from AWC regularly. Only 32.36% of children who were sick were referred by AWW. Satisfaction with quantity and quality of food served was found to be 61% and 45.90% among mothers, respectively. Conclusion: It was concluded that there is a need to improve the quality of services to overcome the problem of under-utilisation of services despite the wide coverage of ICDS services; regular orientation and training courses for AWWs should be conducted to ensure better service delivery to all beneficiaries.

Cytosolic expression of synthetic phytochelatin and bacterial metallothionein genes in Deinococcus radiodurans R1 for enhanced tolerance and bioaccumulation of cadmium.

Due to its exemplary resistance to ionising radiation, oxidative stress, desiccation and several DNA damaging agents, Deinococcus radiodurans R1 (DR1) is considered as one of the most appropriate candidates for the bioremediation of the nuclear waste sites. However, the high sensitivity of this bacterium to heavy metals, which are usually preponderant at nuclear waste dump sites, precludes its application for bioremediation. This study deals with the expression two metal binding peptides in DR1 as an attractive strategy for developing metal tolerance in this bacterium. A synthetic gene (EC20) encoding a phytochelatin analogue with twenty repeating units of glutamate and cysteine was constructed by overlap extension and expressed in DR1. The cyanobacterial metallothionein (MT) gene, smtA was cloned for intracellular expression in DR1. Both the genes were expressed under the native groESL promoter. DR1 strain carrying the recombinant EC20 demonstrated 2.5-fold higher tolerance to Cd(2+) and accumulated 1.21-fold greater Cd(2+) as opposed to the control while the heterologous expression of MT SmtA in DR1 imparted the transformant superior tolerance to Cd(2+) amassing 2.5-fold greater Cd(2+) than DR1 expressing EC20.

Novel 16S rRNA based PCR method targeting Deinococcus spp. and its application to assess the diversity of deinococcal populations in environmental samples.

The members of the genus Deinococcus are extensively studied because of their exemplary radiation resistance. Both ionizing and non-ionizing rays are routinely employed to select upon the radiation resistant deinococcal population and isolate them from the majority of radiation sensitive population. There are no studies on the development of molecular tools for the rapid detection and identification of deinococci from a mixed population without causing the bias of radiation enrichment. Here we present a Deinococcus specific two-step hemi-nested PCR for the rapid detection of deinococci from environmental samples. The method is sensitive and specific to detect deinococci without radiation exposure of the sample. The new protocol was successfully employed to detect deinococci from several soil samples from different geographical regions of India. The PCR method could be adapted to a three-step protocol to study the diversity of the environmental deinococcal population by denaturing gradient gel electrophoresis (DGGE). Sequence analysis of the DGGE bands revealed that the samples harbor diverse populations of deinococci, many of which were not recovered by culturing and may represent novel clades. We demonstrate that the genus specific primers are also suitable for the rapid identification of the bacterial isolates that are obtained from a typical radiation enrichment isolation technique. Therefore the primers and the protocols described in this study can be used to study deinococcal diversity from environmental samples and can be employed for the rapid detection of deinococci in samples or identifying pure culture isolates as Deinococcus species.

Multiple-stress tolerance of ionizing radiation-resistant bacterial isolates obtained from various habitats: correlation between stresses.

Isolation of five ionizing radiation (IR)-resistant bacteria by screening of isolates from various habitats classified as common and stressed is reported. IR-resistant isolates exhibited varying degrees of resistance to gamma-radiation and were classified as highly and moderately radiation resistant. Resistance to ultraviolet (UV) radiation correlated well with gamma-radiation resistance, whereas a comparable desiccation resistance for all the highly and moderately radiation-resistant isolates was observed. However, salt tolerance failed to correlate with IR resistance, indicating a divergent evolution of the salt tolerance and radiation resistance. Characterization of isolates by the amplified rDNA restriction analysis profiling attested to the clustering of these isolates with their stress phenotype. 16S rRNA gene-based analysis of the isolates showed that the bacteria with similar-resistance physiologies clustered together and belonged to related genera. Hydrogen peroxide resistance and mitomycin survival patterns of the isolates indicated the roles of oxidative-stress tolerance in desiccation survival and recombination repair in higher radiation resistance, respectively.