Molecular epidemiology & therapeutic options of carbapenem-resistant Gram-negative bacteria.

Background & objectives: The growing incidence and the wide diversity of carbapenemase-producing bacterial strains is a major concern as only a few antimicrobial agents are active on carbapenem-resistant bacteria. This study was designed to study molecular epidemiology of carbapenem-resistant Gram-negative bacterial (GNB) isolates from the community and hospital settings. Methods: In this study, non-duplicate GNB were isolated from clinical specimens, and phenotypic test such as modified Hodge test, metallo ß-lactamase E-strip test, etc. were performed on carbapenem-resistant bacteria. Multiplex PCR was performed to identify the presence of blaIMP, blaVIM, blaKPC, blaOXA48, blaOXA23, blaSPM, blaGIM, blaSIM and blaNDM. Minimum inhibitory concentration (MIC) of colistin, fosfomycin, minocycline, chloramphenicol and tigecycline was also determined. Results: Of the 3414 GNB studied, carbapenem resistance was 9.20 per cent and maximum resistance (11.2%) was present at tertiary care centre, followed by secondary care (4%) and primary centre (2.1%). Among the carbapenem-resistant bacteria, overall, the most common isolate was Pseudomonas aeruginosa (24%). On multiplex PCR 90.3 per cent carbapenem-resistant isolates were positive for carbapenemase gene. The blaNDM(63%) was the most prevalent gene followed by blaVIM(18.4%). MIC results showed that 88 per cent carbapenem-resistant Enterobacteriaceae were sensitive to fosfomycin, whereas 78 per cent of P. aeruginosa and 85 per cent Acinetobacter spp. were sensitive to colistin. Interpretation & conclusions: Carbapenem resistance in GNB isolates from the community and hospital settings was found to be on the rise and should be closely monitored. In the absence of new antibiotics in pipeline and limited therapeutic options, prudent use of antibiotics and strict infection control practices should be followed in hospital to limit the emergence and spread of multidrug-resistant bacteria.

Evaluation of the Rapidec Carba NP Test Kit for Detection of Carbapenemase-Producing Gram-Negative Bacteria.

Recently, bioMérieux, France, introduced the Rapidec Carba NP test kit for rapid detection of carbapenemase-producing Gram-negative bacteria. This kit was evaluated in this study, and we report sensitivity, specificity, and positive and negative predictive values of 92.6%, 96.2%, 95.83%, and 92.6%, respectively. The test was easy to perform and interpret and relatively inexpensive ($5/Rs 300 per test) and provides a practical solution for early detection of carbapenemase-producing, multidrug-resistant Gram-negative bacteria.

Pesticides-induced biochemical alterations in occupational North Indian suburban population.

Pesticides are used in agriculture to protect crops from insects-pests. Most of the field workers of North Indian population are exposed to commonly used insecticides. In the present study, pesticides induced oxidative stress as well as alterations in the level of acetylcholinesterase (AChE) in a total of 70 male healthy agricultural sprayers, exposed to pesticides for about 5 years, were studied and the results were compared with 70 controls. The levels of antioxidant enzymes (superoxide dismutase, CAT, glutathione-S-transferase and glutathione peroxidase), AChE, lipid peroxidation and glutathione (GSH) contents were determined in their blood erythrocytes (red blood cells (RBCs)). The results indicated significant increase in the levels of malondialdehyde as well as the activities of antioxidant enzymes in pesticide-exposed individuals. The levels of GSH, RBC-AChE activity and plasma antioxidant potential were sharply decreased when compared with control subjects. The ferric-reducing ability of plasma (FRAP) assay was carried out to evaluate the antioxidant potential of pesticide in exposed as well as healthy controls. A significant positive correlation was observed between plasma FRAP value and the activity of AChE from RBCs in pesticides sprayers. Furthermore, these results were supported by enhanced messenger RNA expressions of cytochrome P450 isoform 2E1 (CYP2E1) and gutathione-S-transferase isoform pi (GST-pi) in the white blood cells of the randomly selected pesticide-exposed individuals. The decreased GSH level in human red blood cells accompanied by increase in the levels of the activities of antioxidative enzymes and over expressions of CYP2E1 and GST-pi is an indicative of oxidative stress in pesticides-exposed individuals. The reduced activity of AChE indicates possible occurrence of perturbations in blood as well as neurotoxicity in pesticide sprayers.

Oxidative DNA damage protective activity, antioxidant and anti-quorum sensing potentials of Moringa oleifera.

The aqueous extract of leaf (LE), fruit (FE) and seed (SE) of Moringa oleifera was assessed to examine the ability to inhibit the oxidative DNA damage, antioxidant and anti-quorum sensing (QS) potentials. It was found that these extracts could significantly inhibit the OH-dependent damage of pUC18 plasmid DNA and also inhibit synergistically with trolox, with an activity sequence of LE > FE > SE. HPLC and MS/MS analysis was carried out, which showed the presence of gallic acid, chlorogenic acid, ellagic acid, ferulic acid, kaempferol, quercetin and vanillin. The LE was with comparatively higher total phenolics content (105.04 mg gallic acid equivalents (GAE)/g), total flavonoids content (31.28 mg quercetin equivalents (QE)/g), and ascorbic acid content (106.95 mg/100 g) and showed better antioxidant activity (85.77%), anti-radical power (74.3), reducing power (1.1 ascorbic acid equivalents (ASE)/ml), inhibition of lipid peroxidation, protein oxidation, OH-induced deoxyribose degradation, and scavenging power of superoxide anion and nitric oxide radicals than did the FE, SE and standard alpha-tocopherol. Eventually, LE and FE were found to inhibit violacein production, a QS-regulated behavior in Chromobacterium violaceum 12472.

Polyphenolics from various extracts/fractions of red onion (Allium cepa) peel with potent antioxidant and antimutagenic activities.

In order to determine antioxidant activity, the five extracts/fractions of red onion peel were studied for their total content of phenolics (TPC), flavonoids (TFC), antioxidant activity (AOA), free radical scavenging activity (FRSA), assayed by DPPH radical in the terms of anti-radical power (ARP) and reducing power (RP), expressed as ascorbic acid equivalents (ASE)/ml. High TPC (384.7 +/- 5.0 mg GAE/g), TFC (165.2+/- 3.2 mg QE/g), AOA (97.4 +/- 7.6%), ARP (75.3 +/-4.5) and RP (1.6 +/-0.3 ASE/ml) were found for the ethyl acetate (EA) fraction. EA fraction had markedly higher antioxidant capacity than butylated hydroxytoluene (BHT) in preventive or scavenging capacities against FeCl3-induced lipid peroxidation, protein fragmentation, hydroxyl (site-specific and non-site-specific), superoxide anion and nitric oxide radicals. EA fraction also showed dose dependent antimutagenic activity by following the inhibition of tobacco-induced mutagenicity in Salmonella typhimurium strains (TA102) and hydroxyl radical-induced nicking in plasmid pUC18 DNA. HPLC and MS/MS analysis showed the presence of ferulic, gallic, protocatechuic acids, quercetin and kaempferol. The large amount of polyphenols contained in EA fraction may cause its strong antioxidant and antimutagenic properties. This information shows that EA fraction of red onion peel can be used as natural antioxidant in nutraceutical preparations.

Molecular association between beta-catenin degradation complex and Rac guanine exchange factor DOCK4 is essential for Wnt/beta-catenin signaling.

The canonical Wnt/beta-catenin pathway is a highly conserved signaling cascade that is involved in development and stem cell renewal. The deregulation of this pathway is often associated with increased cell growth and neoplasia. The small GTPase Rac has been shown to influence canonical Wnt signaling by regulating beta-catenin stability through an unknown mechanism. We report that DOCK4, a guanine nucleotide exchange factor (GEF) for Rac and a member of the CDM family of unconventional GEFs, mediates Wnt-induced Rac activation in the canonical Wnt/beta-catenin pathway. DOCK4 expression regulates cellular beta-catenin levels in response to the Wnt signal, in vitro. Biochemical studies demonstrate that DOCK4 interacts with the beta-catenin degradation complex, consisting of the proteins adenomatosis polyposis coli, Axin and glycogen synthase kinase 3beta (GSK3beta). This molecular interaction enhances beta-catenin stability and Axin degradation. Furthermore, we observe that DOCK4 is phosphorylated by GSK3beta, which enhances Wnt-induced Rac activation. Using a T-cell factor reporter zebrafish we confirm that DOCK4 is required for Wnt/beta-catenin activity, in vivo. These results elucidate a novel intracellular signaling mechanism in which a Rac GEF, DOCK4 acts as a scaffold protein in the Wnt/beta-catenin pathway.

Lead and cadmium concentration in the seminal plasma of men in the general population: correlation with sperm quality.

The concentration of lead and cadmium in the seminal plasma of men in the general population, including fertile and infertile subjects, was measured. Semen samples were categorised as demonstrating oligospermia (sperm concentration < 20 x 10(6)/ml), asthenospermia (<50% motilesperm), oligoasthenospermia (a combination of the two criteria), and azoospermia (no sperm). An increase in lead and cadmium levels was observed in infertile men and there was a significant negative correlation of cadmium and lead semen concentration with sperm motility and sperm concentration in oligoasthenospermic men.

Cultivation of Mycobacterium tuberculosis from pleural tissue and its histopathology in suspected cases of tuberculous pleural effusion.

The majority of idiopathic pleural effusion are considered to be of tuberculous etiology. The culture of bacilli provide most authentic evidence of disease process. This study was carried out on 21 cases of suspected tuberculous pleural effusion. The pleural fluid and tissue was subjected for smear examination, and cultivation of tubercle bacilli along with pleural biopsy. Overall the diagnosis could be made out of 8 (38.09%) cases. The cultivation of Mycobacterium tuberculosis from pleural tissue was more sensitive (33.33%) as compared to histopathology (19.05%). However, the triad of cultivation of Mycobacterium tuberculosis from pleural tissue, pleural histopathology and pleural fluid smear examination or culture should be done in each suspected case of tuberculous effusion.