Leptospira interrogans outer membrane protein-based nanohybrid sensor for the diagnosis of leptospirosis

Leptospirosis is an underestimated tropical disease caused by the pathogenic Leptospira species and responsible for several serious health problems. Here, we aimed to develop an ultrasen-sitive DNA biosensor for the rapid and on-site detection of the Loa22 gene of Leptospira interrogans using a gold nanoparticle–carbon nanofiber composite (AuN/CNF)-based screen-printed electrode. Cyclic voltammetry and electrochemical impedance were performed for electrochemical analysis. The sensitivity of the sensor was 5431.74 µA/cm2 /ng with a LOD (detection limit) of 0.0077 ng/µL using cyclic voltammetry. The developed DNA biosensor was found highly specific to the Loa22 gene of L. interrogans, with a storage stability at 4¿ C for 180 days and a 6% loss of the initial response. This DNA-based sensor only takes 30 min for rapid detection of the pathogen, with a higher specificity and sensitivity. The promising results obtained suggest the application of the developed sensor as a point of care device for the diagnosis of leptospirosis.This research was funded by the Indian Council of Medical Research (ICMR), grant Leptos/27/2013-ECD-1. N.C.-M. acknowledges the Portuguese Foundation for Science and Technology under the Horizon 2020 Program (PTDC/PSI-GER/28076/2017), Supported by Excellence project FIM UHK (K.K.)

An HIV1/2 point of care test on sputum for screening TB/HIV co-infection in central India – Will it work?

AbstractObjectiveTo determine whether the OraQuick® HIV-1/2 Assay (OraSure Technologies, Inc., Bethlehem, PA, USA) in sputum is a valid tool for HIV surveillance among TB patients.MethodsA cross sectional study was carried out on sputa of patients diagnosed with tuberculosis. Sputa were tested for antibodies to HIV using OraQuick® HIV-1/2 Assay (OraSure Technologies, Inc., Bethlehem, PA, USA). The results were compared with results of serum ELISA.ResultsCompared to serum ELISA, the OraQuick® HIV-1/2 Assay in sputum specimens reported 90% sensitivity (9/10) and 100% specificity (307/307), with a positive predictive value of 100% (95% CI: 66.37%–100.00%) and a negative predictive value of 99.68% (95% CI: 98.20%–99.99%).ConclusionsThis testing method may provide a useful strategy for conducting HIV surveillance in possible co-infected TB patients at peripheral centres. Since there is no investment on infrastructure, it may be possible for paramedical health professionals to carry out the test, particularly in areas with low HIV endemicity

Synthesis of ethyl oleate employing synthetic hydrogel-immobilized lipase of Bacillus coagulans MTCC-6375

Ten polymeric hydrogels were chemically synthesized by varying the concentrations of copolymer (DMA) and cross-linker (MBAm) molecules. An alkaline lipase of Bacillus coagulans MTCC-6375 was immobilized onto a poly (MAc-co-DMA-cl-MBAm)-hydrogel support at pH 8.5 and temperature 55&ordm;C in 16 h. The bound lipase possessed 7.6 U.g⁻&sup1; (matrix) lipase activity with a specific activity of 18 U.mg⁻&sup1; protein. Hydrogel bound-lipase catalyzed esterification of oleic acid and ethanol to synthesize ethyl oleate in n-nonane. Various kinetic parameters were optimized to produce ethyl oleate using immobilized lipase. The optimal parameters were bound enzyme/substrate (E/S) ratio 0.62 mg/mM, ethanol/oleic acid 100 mM:75 mM or 100 mM:100 mM, incubation time 18 h and reaction temperature 55&ordm;C that resulted in approximately 53% conversion of reactants into ethyl oleate in n-nonane. However, addition of a molecular sieve to the reaction mixture promoted the conversion to 58% in 18 h in n-nonane, which was equivalent to 55 mM of ethyl oleate produced.<br /

Development of Novel Interspecific Fertile Cytotype (4X) Of \u3cem\u3ePennisetum glaucum\u3c/em\u3e X \u3cem\u3ePennisetum purpureum\u3c/em\u3e Utilizing Modified Ploidy Coupled With Embryo Rescue Technique

Interspecific hybrids of genus Pennisetum (P. glaucum x P. purpureum) is the one of the most popular manmade hybrid. It combines the unique features of both P. glaucum (Pearl millet; Bajra) and P. purpureum (Napier; Elephant grass) species, which makes it more resilient to harsh environments with superior fodder quality. Due to ploidy level variation among the parents, these hybrids are sterile and propagated vegetatively only. To overcome this, attempts were made in the present study by exploring the feasibility of novel tetraploid pearl millet (2n=4x=28; Tetra 1; INGR 09047) developed at IGFRI, as a female parent in crossing program involving different Napier genotypes as male parent. Due to limited crossability and hybrid necrosis issues among countless crosses (\u3e 1000), only 1% seed set was initially recorded that too in shriveled state and the developing embryos were aborted after 10-14 days of pollination and fertilization. To save these, embryo rescue technique was standardized and the developing embryos were dissected out aseptically and rescued after 8-10 days of pollination. Continuous crossing programme along with screening of large tissue culture raised nurseries resulted in development of a novel tetraploid seed producing BN hybrid (TBN-20-15) along with 14 novel sterile tetraploid BN hybrids. Presence of univalent chromosomes leads to sterility while proper pairing between parents of TBN-20-15 hybrid have significant effect on fertility. The fertile hybrid is able to produce \u3e15,000-20,000 seeds throughout the year with 80-90% seed germination ability. Their hybridity was confirmed by morphology, molecular and cytogenetic studies. This fertile tetraploid BN hybrid (TBN-20-15) reported for the first time globally will be very helpful in easy and cost-effective dissemination of this highly potential forage crop to the farmer’s field. It has the potential to be the game changer in biofuel production, grassland rejuvenation programs besides bridging the fodder demand supply deficit

The Cellular Stress Response Interactome and Extracellular Matrix Cross-Talk during Fibrosis: A Stressed Extra-Matrix Affair

Diverse internal and external pathologic stimuli can trigger cellular stress response pathways (CSRPs) that are usually counteracted by intrinsic homeostatic machinery, which responds to stress by initiating complex signaling mechanisms to eliminate either the stressor or the damaged cells. There is growing evidence that CSRPs can have context-dependent homeostatic or pathologic functions that may result in tissue fibrosis under persistence of stress. CSRPs can drive intercellular communications through exosomes (trafficking and secretory pathway determinants) secreted in response to stress-induced proteostasis rebalancing. The injured tissue environment upon sensing the stress turns on a precisely orchestrated network of immune responses by regulating cytokine-chemokine production, recruitment of immune cells, and modulating fibrogenic niche and extracellular matrix (ECM) cross-talk during fibrotic pathologies like cardiac fibrosis, liver fibrosis, laryngotracheal stenosis, systemic scleroderma, interstitial lung disease and inflammatory bowel disease. Immunostimulatory RNAs (like double stranded RNAs) generated through deregulated RNA processing pathways along with RNA binding proteins (RBPs) of RNA helicase (RNA sensors) family are emerging as important components of immune response pathways during sterile inflammation. The paradigm-shift in RNA metabolism associated interactome has begun to offer new therapeutic windows by unravelling the novel RBPs and splicing factors in context of developmental and fibrotic pathways. We would like to review emerging regulatory nodes and their interaction with CSRPs, and tissue remodeling with major focus on cardiac fibrosis, and inflammatory responses underlying upper airway fibrosis

Multiple Analytical Approaches Reveal Distinct Gene-Environment Interactions in Smokers and Non Smokers in Lung Cancer

Complex disease such as cancer results from interactions of multiple genetic and environmental factors. Studying these factors singularly cannot explain the underlying pathogenetic mechanism of the disease. Multi-analytical approach, including logistic regression (LR), classification and regression tree (CART) and multifactor dimensionality reduction (MDR), was applied in 188 lung cancer cases and 290 controls to explore high order interactions among xenobiotic metabolizing genes and environmental risk factors. Smoking was identified as the predominant risk factor by all three analytical approaches. Individually, CYP1A1*2A polymorphism was significantly associated with increased lung cancer risk (OR = 1.69;95%CI = 1.11–2.59,p = 0.01), whereas EPHX1 Tyr113His and SULT1A1 Arg213His conferred reduced risk (OR = 0.40;95%CI = 0.25–0.65,p<0.001 and OR = 0.51;95%CI = 0.33–0.78,p = 0.002 respectively). In smokers, EPHX1 Tyr113His and SULT1A1 Arg213His polymorphisms reduced the risk of lung cancer, whereas CYP1A1*2A, CYP1A1*2C and GSTP1 Ile105Val imparted increased risk in non-smokers only. While exploring non-linear interactions through CART analysis, smokers carrying the combination of EPHX1 113TC (Tyr/His), SULT1A1 213GG (Arg/Arg) or AA (His/His) and GSTM1 null genotypes showed the highest risk for lung cancer (OR = 3.73;95%CI = 1.33–10.55,p = 0.006), whereas combined effect of CYP1A1*2A 6235CC or TC, SULT1A1 213GG (Arg/Arg) and betel quid chewing showed maximum risk in non-smokers (OR = 2.93;95%CI = 1.15–7.51,p = 0.01). MDR analysis identified two distinct predictor models for the risk of lung cancer in smokers (tobacco chewing, EPHX1 Tyr113His, and SULT1A1 Arg213His) and non-smokers (CYP1A1*2A, GSTP1 Ile105Val and SULT1A1 Arg213His) with testing balance accuracy (TBA) of 0.6436 and 0.6677 respectively. Interaction entropy interpretations of MDR results showed non-additive interactions of tobacco chewing with SULT1A1 Arg213His and EPHX1 Tyr113His in smokers and SULT1A1 Arg213His with GSTP1 Ile105Val and CYP1A1*2C in nonsmokers. These results identified distinct gene-gene and gene environment interactions in smokers and non-smokers, which confirms the importance of multifactorial interaction in risk assessment of lung cancer

External Jugular Vein Aneurysm with Thrombus Presenting as Painful Neck Mass: A Case Report

Abstract External jugular vein aneurysm with thrombosis presenting as neck swelling is a rare clinical entity and rarely encountered in routine clinical practice. We present a case of a 45-year-old female with external jugular vein aneurysm presenting as a painful lump in the neck. Ultrasound of the neck and CT angiography showed saccular dilation of the lower end of the external jugular vein with thrombosis within the aneurysm. Saccular aneurysm of the external jugular vein is very uncommon and can lead to thrombotic complications with serious consequence