Diagnosis of pleural tuberculosis by multi-targeted loop-mediated isothermal amplification assay based on SYBR Green I reaction: comparison with GeneXpert® MTB/RIF assay.

BACKGROUND: Diagnosis of pleural tuberculosis (TB) is tedious owing to its close resemblance with malignant pleural effusion and sparse bacterial load in clinical specimens. There is an immediate need to design a rapid and dependable diagnostic test to prevent unnecessary morbidity/mortality. RESEARCH DESIGN AND METHODS: A multi-targeted loop-mediated isothermal amplification (MT-LAMP) was deliberated using mpt64 and IS6110 to diagnose pleural TB within pleural fluids/biopsies. MT-LAMP products were analyzed by gel-based and visual detection methods, viz. SYBR Green I, SYBR Green I+deoxyuridine triphosphate uracil-N-glycosylase (dUTP-UNG), and dry methyl green reactions. RESULTS: In a pilot study, while assessing pleural TB/non-TB control subjects (n = 40), both SYBR Green I+dUTP-UNG/gel-based MT-LAMP assays exhibited better sensitivity/specificity than SYBR Green I and dry methyl green MT-LAMP. Since it is facile to work with SYBR Green I+dUTP-UNG than gel-based MT-LAMP, we validated the performance of SYBR Green I+dUTP-UNG in a higher number of specimens (n = 97), which revealed somewhat higher sensitivity (85.2 vs. 81.5%) and specificity (97.7 vs. 90.7%) than SYBR Green I MT-LAMP. Furthermore, the sensitivity attained by SYBR Green I+dUTP-UNG MT-LAMP was significantly higher (p < 0.001) than GeneXpert. CONCLUSIONS: Our SYBR Green I+dUTP-UNG MT-LAMP is a simple and reliable method to diagnose pleural TB, which may translate into a point-of-care test.

Groundwater hydro-geochemistry, quality, microbiology and human health risk assessment in semi-arid area of Rajasthan, India: a chemometric approach.

The present investigation focused on groundwater hydro-geochemistry of Alsisar block of Jhunjhunu district, India, aims on evaluating the quality of groundwater for drinking and irrigation purposes and assessing the human health risk from ingestion of groundwater. The groundwater of Alsisar block is neutral to alkaline, brackish and very hard in nature. Total dissolved solids, total hardness, Na+, Mg2+, HCO3-, F- and NO3- in majority of the groundwater samples were exceeding the World Health Organization and Bureau of Indian Standards recommended limits. The drinking water quality index ranged from 111.53 to 492.84. None of the sample belonged to excellent and good categories of drinking water quality. Fluoride varied from 0.018 to 4.176 mg L-1, and nitrate varied from 0.34 to 520.66 mg L-1 in groundwater. The non-carcinogenic risk assessment for children, men and women owing to ingestion of fluoride and nitrate-enriched groundwater indicates human health risks in the entire study area. Irrigation with groundwater of Alsisar block is liable to cause salinity and magnesium hazard to agricultural crops grown in the area. Source apportionment using principal component analysis suggests the geogenic origin of fluoride and anthropogenic origin of nitrate. Na+-Mg2+-Cl- followed by Na+-Mg2+-HCO3- are the predominant hydrochemical facies in the groundwater of Alsisar block. Silicate rock weathering, ion exchange and evaporation are the predominating processes governing ionic concentrations in the groundwater. Biochemical and molecular tests demonstrated the presence of Brevibacillus borstelensis strain DSM 6347 16s rRNA and Bacillus paramycoides strain MCCC 1A04098 16s rRNA in the groundwater of the area.

Synthesis, Spectral Characterization, Antibacterial and Anticancer Activity of some Titanium Complexes.

BACKGROUND: After the discovery of cisplatin, first non platinum anticancer drugs having excellent efficacy were budotitane and TiCl2(cp)2 but action mechanism is not clear. Therefore, we hereby reporting synthesis and biological activities novel titanium complexes to explore their mode of action. OBJECTIVES: Synthesis, spectral characterization, antibacterial and anticancer activity of some titanium complexes. Antibacterial studies on various bacterial strains and anticancer studies on HeLa, C6, CHO cancerous cell lines have been performed. Further, the cell death mechanistic study was done on CHO cell lines. METHOD: Titanium complexes with and without labile groups have been synthesized by reacting of TiCl4 with nitrogen containing ligands viz. 1,2-diaminocyclohexane, 1,10-Phenanthroline, adamantylamine, 2,2'-bipyridine, 4,4'-dimethyl-2,2'-bipyridine in predetermined molar ratios. Antibacterial and anticancer studies were performed by agar well diffusion method and MTT assay respectively. Cell cycle analysis is done by using flow cytometry. RESULTS: Complex 2 i.e TiCl2(Phen)2 showed better activity than other complexes as an antibacterial as well as anticancer agent. Phase contrast imaging indicates that observed morphological changes of cells was dose dependent. Cell death mechanistic study have shown the increase in sub G0 phase population as well as formation of blebbing and fragmentation of chromatin material which is an indicative measure of apoptosis. CONCLUSION: Complex 2 proved to be more effective bactericide and cytotoxic agent. Cell cycle analysis showed cell arrest in G0 phase. Apoptosis percentage was found to increase in a dose dependent manner. So, prepared titanium complexes can be put to use as an important chemotherapeutic agents.