Prevalence of Naegleria fowleri in Environmental Samples from Northern Part of India

Naegleria fowleri the causative agent of Primary Amoebic Meningoencephalitis, is ubiquitously distributed worldwide in various warm aquatic environments and soil habitats. The present study reports on the presence of Naegleria spp. in various water bodies present in Rohtak and Jhajjar district, of state Haryana, India. A total of 107 water reservoirs were screened from summer till autumn (2012 and 2013). In order to isolate Naegleria spp. from the collected water samples, the water samples were filtered and the trapped debris after processing were transferred to non-nutrient agar plates already seeded with lawn culture of Escherichia coli. Out of total 107 water samples, 43 (40%) samples were positive by culture for free living amoeba after incubation for 14 days at 37°C. To identify the isolates, the ITS1, 5.8SrDNA and ITS2 regions were targeted for PCR assay. Out of total 43 positive samples, 37 isolates were positive for Naegleria spp. using genus specific primers and the most frequently isolated species was Naegleria australiensis. Out of 37 Naegleria spp. positive isolates, 1 isolate was positive for Naegleria fowleri. The sequence analysis revealed that the Naegleria fowleri strain belonged to Type 2.     

Fabrication of conducting electrospun nanofibers scaffold for three-dimensional cells culture

Electrospinning is a versatile method to fabricate nanofibers of a range of polymeric and composite materials suitable as scaffolds for tissue engineering applications. In this study, we report the fabrication and characterization of polyaniline-carbon nanotube/poly(N-isopropyl acrylamide-co-methacrylic acid) (PANI-CNT/PNIPAm-co-MAA) composite nanofibers and PNIPAm-co-MAA nanofibers suitable as a three-dimensional (3D) conducting smart tissue scaffold using electrospinning. The chemical structure of the resulting nanofibers was characterized with FTIR and (1)H NMR spectroscopy. The surface morphology and average diameter of the nanofibers were observed by SEM. Cellular response of the nanofibers was studied with mice L929 fibroblasts. Cell viability was checked on 7th day of cell culture by double staining the cells with calcein-AM and PI dye. PANI-CNT/PNIPAm-co-MAA composite nanofibers were shown the highest cell growth and cell viability as compared to PNIPAm-co-MAA nanofibers. Cell viability in the composite nanofibers was obtained in order of 98% that indicates the composite nanofibers provide a better environment as a 3D scaffold for the cell proliferation and attachment suitable for tissue engineering applications.     

Impact of Endobronchial Ultrasound (EBUS) Training on the Diagnostic Yield of Conventional Transbronchial Needle Aspiration for Lymph Node Stations 4R and 7

BackgroundThere is sparse literature on whether training in endobronchial ultrasound (EBUS)-guided transbronchial needle aspiration (TBNA) improves the diagnostic yield of conventional TBNA (cTBNA).ObjectivesThe aim of this study was to evaluate the diagnostic yield of cTBNA before and after the introduction of EBUS.MethodsThis was a retrospective analysis of patients who underwent cTBNA at our center. The study was divided into two periods, before and after the introduction of EBUS at our facility. The diagnostic yield of cTBNA was compared between the study periods. Rapid on-site cytological examination was not available.ResultsA total of 1,050 patients (61.6% men; mean age 45.6 years) underwent cTBNA during the study period (849 before EBUS; 201 after EBUS). Sarcoidosis (n = 527) followed by bronchogenic carcinoma (n = 222) formed the most common indications for performing cTBNA. There was a significant increase in both the success of obtaining a representative sample (from 71% to 85%), and the diagnostic yield (from 33% to 49.5%) of cTBNA, after the introduction of EBUS. The increase in the diagnostic yield of cTBNA after introduction of EBUS remained significant even after adjusting for years of performing cTBNA and the type of anesthesia (topical vs. sedation and topical) on a multivariate analysis.ConclusionThe diagnostic yield of cTBNA at our facility increased after the introduction of EBUS-TBNA. However, given the retrospective nature of the study, prospective studies are required to confirm our findings.     

Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulans BTS-3

A thermophilic isolate Bacillus coagulans BTS-3 produced an extracellular alkaline lipase, the production of which was substantially enhanced when the type of carbon source, nitrogen source, and the initial pH of culture medium were consecutively optimized. Lipase activity 1.16 U/ml of culture medium was obtained in 48 h at 55 degrees C and pH 8.5 with refined mustard oil as carbon source and a combination of peptone and yeast extract (1:1) as nitrogen sources. The enzyme was purified 40-fold to homogeneity by ammonium sulfate precipitation and DEAE-Sepharose column chromatography. Its molecular weight was 31 kDa on SDS-PAGE. The enzyme showed maximum activity at 55 degrees C and pH 8.5, and was stable between pH 8.0 and 10.5 and at temperatures up to 70 degrees C. The enzyme was found to be inhibited by Al3+, Co2+, Mn2+, and Zn2+ ions while K+, Fe3+, Hg2+, and Mg2+ ions enhanced the enzyme activity; Na+ ions have no effect on enzyme activity. The purified lipase showed a variable specificity/hydrolytic activity towards various 4-nitrophenyl esters.