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Humans are exposed to a cocktail of heavy metal toxicants at the same time in the environment rather than single metal. The kidney is often a site of early damage due to high renal contact to these pollutants. This study was done to examine the cumulative toxic effect of multiple elements prevalent in the environment. To explore the effect of subchronic exposure to heavy metal mixture male and female Swiss albino mice were randomly divided into 14 groups and given varying doses [MPL (maximum permissible limit), 1X, 5X, 10X, 50X, or 100X] of the multiple metals and metalloid mixtures via drinking water for 8 weeks. It was determined that metal treatment caused increased metal load in renal tissue. The kidney function deteriorated in response to 10X, 50X, 100X concentration of the dosing mixture was found associated to oxidative stress, glomerular damage, necrosis, cell death and further exacerbation of the inflammation.

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Multidrug-resistant tuberculosis (MDR-TB) requires treatment with fluoroquinolone (FLQ) drugs, however, the excessive use of FLQ has led to the rise of extensively drug-resistant TB. In 2019, ~ 20% of total MDR-TB cases were estimated to be resistant to FLQ drugs. In the present study, we developed and evaluated the utility of high-resolution melt curve analysis (HRM) for the rapid detection of FLQ-resistant Mycobacterium tuberculosis for the first time directly from sputum samples. A reference plasmid library was generated for the most frequently observed mutations of gyrA gene and was used to discriminate between mutant and wild-type samples in the FLQ-HRM assay. The developed assay was evaluated on n = 25 MDR M. tuberculosis clinical isolates followed by validation on archived sputum DNA (n = 88) using DNA sequencing as a gold standard. The FLQ-HRM assay showed a 100% sensitivity [95% Confidence Interval (CI): 71.5 to 100] and specificity (95% CI: 39.7 to 100) in smear-positive category, and a sensitivity of 88.9% (95% CI: 77.3 to 95.8) with 84.2% (95% CI: 60.4 to 96.6) specificity in smear-negative category. The assay showed a high level of concordance of ~ 90% (κ = 0.74) with DNA sequencing, however, we were limited by the absence of phenotypic drug susceptibility testing data. In conclusion, HRM is a rapid, cost-effective (INR 150/USD 1.83) and closed-tube method for direct detection of FLQ resistance in sputum samples including direct smear-negative samples.

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Paenibacillus sp. strain GLM-08 was isolated from a lignite mine waste site in the Barmer basin, Rajasthan, India. The strain is efficient in heterotrophic nitrification and aerobic denitrification. This bacterium could remove approximately more than 95% of NH4+, NO3-, and NO2- in 24 h. The average nitrogen (N) removal rate of the strain was found to be 4.775 mg/L/H, 5.66 mg/L/H, and 5.01 mg/L/H for NH4+, NO3-, and NO2-, respectively. Bioaugmentation of mine wastewater with Paenibacillus sp. strain GLM-08 demonstrated N removal of 86.6% under conditions of a high load of NH4+. The presence of potential genetic determinants (nxrB, nirS, and nosZ) having role in heterotrophic nitrification and aerobic denitrification was confirmed by PCR based analysis. The findings show that this bacterium performs simultaneous nitrification and denitrification and has a high nitrogen removal efficiency indicating the potential application of the strain in the treatment of wastewater.

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In 2019, amongst half a million new rifampicin-resistant tuberculosis (TB) cases, 78% were multi drug-resistant TB (MDR-TB). Access to rapid and Universal-Drug susceptibility testing (DST) to patients in remote areas is a major challenge to combat drug-resistant TB. To overcome this challenge, we had recently reported the development of 'TB Concentration & Transport kit' for bio-safe ambient temperature transport of dried sputum on filter-paper (Trans-Filter). The present study was conducted to evaluate the utility of DNA extracted from sputum on Trans-Filter in a Multiplex PCR-based sequencing assay (Mol-DSTseq) for diagnosing drug-resistant TB. The developed Mol-DSTseq assays were standardized on Mycobacterium tuberculosis clinical isolates (n = 98) and further validated on DNA extracted from sputum on Trans-Filter (n = 100). Using phenotypic DST as gold standard, the Mol-DSTseq assay showed 100% (95% Confidence Interval [CI] 79.4-100%) and 73.3% (95% CI 54.1-87.7%) sensitivity for detecting rifampicin and isoniazid resistance with a specificity of 85.1% (95% CI 66.2-95.8%) and 100% (95% CI:82.3-100%), respectively. For fluoroquinolones and aminoglycosides, the Mol-DSTseq assay showed a sensitivity of 78.5% (95% CI 49.2-95.3%) and 66.6% (95% CI 9.4-99.1%) with a specificity of 88.2% (95% CI 72.5-96.7%) and 100% (95% CI 93.1-100%), respectively. The Mol-DSTseq assays exhibited a high concordance of ~ 83-96% (κ value: 0.65-0.81) with phenotypic DST for all drugs. In conclusion, the 'TB Concentration and Transport kit' was compatible with Mol-DSTseq assays and has the potential to provide 'Universal-DST' to patients residing in distant areas in high burden countries, like India for early initiation of anti-tubercular treatment.

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The gut microbiota has a vital role in the maintenance of intestinal homeostasis. Several studies have revealed that environmental exposure to pollutants such as heavy metals may contribute to the progression of extensive list of diseases which may further lead to perturbations in the gut leading to dysbiosis. This manuscript critically reviews the alterations in the gut microbiota composition and function upon exposure to various toxic heavy metals prevalent in the environment. The disturbance in gut microbial ecology also affects the microbial metabolic profile which may alter the speciation state and bioavailability heavy metals thus affecting metal uptake-absorption/detoxification mechanisms associated to heavy metal metabolism. The toxic effects of various heavy metals either in single or in multimetallic combination and the gut microbiota associated host health and disease condition need a comprehensive assessment with important consideration for therapeutic and protective strategies against the damage to gut microbiota.

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Hydrocarbon is a primary source of energy in the current urbanized society. Considering the increasing demand, worldwide oil productions are declining due to maturity of oil fields and because of difficulty in discovering new oil fields to substitute the exploited ones. To meet current and future energy demands, further exploitation of oil resources is highly required. Microorganisms inhabiting in these areas exhibit highly diverse catabolic activities to degrade, transform, or accumulate various hydrocarbons. Enrichment of hydrocarbon-utilizing bacteria in oil basin is caused by continuous long duration and low molecular weight hydrocarbon microseepage which plays a very important role as an indicator for petroleum prospecting. The important microbial metabolic processes in most of the oil reservoir are sulfate reduction, fermentation, acetogenesis, methanogenesis, NO3- reduction, and Fe (III) and Mn (IV) reduction. The microorganisms residing in these sites have critical control on petroleum composition, recovery, and production methods. Physical characteristics of heavy oil are altered by microbial biotransformation and biosurfactant production. Considering oil to be one of the most vital energy resources, it is important to have a comprehensive understanding of petroleum microbiology. This manuscript reviews the recent research work referring to the diversity of bacteria in oil field and reservoir sites and their applications for enhancing oil transformation in the target reservoir and geomicrobial prospecting scope for petroleum exploration.

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BACKGROUND: Quadratus lumborum block (QLB) provides somatic and visceral analgesia to the lower thoracic and abdominal wall. The aim was to investigate the analgesic effect of dexamethasone with levobupivacaine in QLB in patients undergoing unilateral inguinal hernia repair surgery. METHODS: A total of 90 patients of American Society of Anaesthesiologists (ASA) I/II were randomly divided into two groups. Group L received 0.25% levobupivacaine (20 ml) + normal saline (1 ml) and group D received 0.25% levobupivacaine (20 ml) + 4 mg dexamethasone (1 ml) in QL plane on the operated side using ultrasound, after completion of surgery under spinal anaesthesia. The primary objective was to compare time for first rescue analgesia. The secondary objectives were total rescue analgesic consumption and numeric rating scale (NRS) in the first 24 h. RESULTS: The demographic data age, sex, height, weight and ASA were comparable in both groups. The mean time to request for first rescue analgesia was longer in group D compared to group L (1016.02 ± 205.97 min versus 640 ± 132.96 min; P < 0.0001). The mean total tramadol consumption in the first 24 h was lower in group D compared to group L (233.55 ± 86.92 mg versus 328.22 ± 78.74 mg; P < 0.0001). Patients in group D had significantly lower NRS scores at rest and on movement as compared to group L. CONCLUSIONS: The addition of dexamethasone to levobupivacaine in QLB results in prolonged duration of postoperative analgesia, less rescue analgesic requirements and better quality of analgesia as compared to levobupivacaine in unilateral inguinal hernia repair surgery.

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A uranium (U)-resistant and -accumulating Pseudomonas aeruginosa strain was characterized to assess the response of toxic metals toward its growth and expression of metal resistance determinants. The bacterium showed MIC (minimum inhibitory concentration) values of 6, 3, and 2 mM for Zn, Cu, and Cd, respectively; with resistance phenotype conferred by periplasmic Cu sequestering copA and RND type heavy metal efflux czcA genes. Real-time PCR-based expression analysis revealed significant upregulation of both these genes upon exposure to low concentrations of metals for short duration, whereas the global stress response gene sodA encoding superoxide dismutase enzyme was upregulated only at higher metal concentrations or longer exposure time. It could also be inferred that copA and czcA are involved in providing resistance only at low metal concentrations, whereas involvement of "global stress response" phenomenon (expression of sodA) at higher metal concentration or increased exposure was evident. This study provides significant understanding of the adaptive response of bacteria surviving in metal and radionuclide contaminated environments along with the development of real-time PCR-based quantification method of using metal resistance genes as biomarker for monitoring relevant bacteria in such habitats.

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OBJECTIVE: To evaluate serum vascular endothelial growth factor (VEGF) levels in children with acute lymphoblastic leukemia (ALL) during the induction phase of chemotherapy. DESIGN: Prospective sudy. SETTING: Setting: Hospital-based study over 18 months. PATIENTS: 30 children with ALL and 17 healthy age- and sex-matched controls. INTERVENTION: Serum concentration of VEGF-A-165 isoform (s-VEGF) was measured by enzyme-linked immunoabsorbant assay at diagnosis and at the end of induction chemotherapy. MAIN OUTCOME MEASURES: s-VEGF was compared with markers of tumor burden. Kinetics of s-VEGF was assessed in response to induction chemotherapy. RESULTS: Median VEGF was significantly lower in untreated patients than in controls (17.0 vs. 42.6 pg/mL, P=0.004). s-VEGF levels were fairly correlated with WBC count (r=-0.56, P=0.004) and LDH (r=-0.52, P=0.02) at diagnosis. All patients but one were in morphologic remission at the end of induction. Median s-VEGF concentration on day 29/33 was significantly higher than on day 1 (44.2 pg/mL, P=0.009). CONCLUSIONS: Untreated children with ALL have significantly lower s-VEGF concentration than controls. At the end of the induction therapy, s-VEGF increased to levels similar to controls. The role of ligand-receptor interaction between VEGF and VEGF receptors on leukemia cells needs to be further delineated.To identify the risk factors for scholastic backwardness in children.

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Ten bacterial strains isolated from uranium mine wastes were characterized in terms of their uranium and other metal resistance and accumulation. 16S rRNA gene sequence analysis identified the strains as members of genera Bacillus, Serratia, and Arthrobacter. Strains were able to utilize various carbon sources, particularly aromatic hydrocarbons, grow at broad pH and temperature ranges and produce non specific acid phosphatase relevant for metal phosphate precipitation in contaminated environment. The isolates exhibited high uranium and other heavy metals (Ni, Co, Cu and Cd) resistance and accumulation capacities. Particularly, Arthrobacter sp. J001 and Bacillus sp. J003 were superior in terms of U resistance at low pH (pH 4.0) along with metals and actinides (U and Th) removal with maximum cell loading of 1088 µmol U, 1293 µmol Th, 425 µmol Cu, 305 µmol Cd, 377 µmol Zn, 250 µmol Ni g(-1) cell dry wt. Genes encoding P(1B)-type ATPases (Cu-CPx and Zn-CPx) and ABC transporters (nik) as catalytic tools for maintaining cellular metal homeostasis were detected within several Bacillus spp., with possible incidence of horizontal gene transfer for the later gene showing phylogenetic lineage to α Proteobacteria members. The study provides evidence on intrinsic abilities of indigenous bacteria from U-mine suitable for survival and cleaning up of contaminated mine sites.

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Uranium biomineralization by a metal-resistant Pseudomonas aeruginosa strain isolated from uranium mine waste was characterized for its potential in bioremediation. Uranium resistance, its cellular localization and chemical nature of uranium-bacteria interaction were elucidated. Survival and uranium biomineralization from mine water were investigated using microcosm experiments. The selected bacterium showed U resistance and accumulation (maximum of 275 mg U g(-1)cell dry wt.) following incubation in 100 mg U L(-1), pH 4.0, for 6 h. Transmission electron microscopy and X-ray diffraction analyses revealed that bioaccumulated uranium was deposited within the cell envelope as needle shaped U-phosphate compounds that attain crystallinity only at pH 4.0. A synergistic involvement of deprotonated phosphate and carboxyl moieties in facilitating bioprecipitation of uranium was evident from FTIR analysis. Based on these findings we attribute the localized U sequestration by this bacterium as innocuous complex to its possible mechanism of uranium resistance. Microcosm data confirmed that the strain can remove soluble uranium (99%) and sequester it as U oxide and phosphate minerals while maintaining its viability. The study showed that indigenous bacteria from contaminated site that can survive uranium and other heavy metal toxicity and sequester soluble uranium as biominerals could play important role in uranium bioremediation.

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Heavy metal sequestration by a multimetal resistant Pseudomonas strain isolated from a uranium mine was characterized for its potential application in metal bioremediation. 16S rRNA gene analysis revealed phylogenetic relatedness of this isolate to Pseudomonas fluorescens. Metal uptake by this bacterium was monophasic, fast saturating, concentration and pH dependent with maximum loading of 1048 nmol Ni(2+) followed by 845 nmol Co(2+), 828 nmol Cu(2+) and 700 nmol Cd(2+)mg(-1) dry wt. Preferential metal deposition in cell envelope was confirmed by TEM and cell fractionation. FTIR spectroscopy and EDX analysis revealed a major role of carboxyl and phosphoryl groups along with a possible ion exchange mechanism in cation binding. Binary system demonstrated selective metal binding affinity in the order of Cu(2+)>Ni(2+)>Co(2+)>Cd(2+). A comparison with similar metal uptake reports considering live bacteria strongly indicated the superiority of this strain in metal sequestration, which could be useful for developing efficient metal removal system.

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