Discovery of new mutually orthogonal bioorthogonal cycloaddition pairs through computational screening.

Density functional theory (DFT) calculations and experiments in tandem led to discoveries of new reactivities and selectivities involving bioorthogonal sydnone cycloadditions. Dibenzocyclooctyne derivatives (DIBAC and BARAC) were identified to be especially reactive dipolarophiles, which undergo the (3+2) cycloadditions with N-phenyl sydnone with the rate constant of up to 1.46 M-1 s-1. Most signifcantly, the sydnone-dibenzocyclooctyne and norbornene-tetrazine cycloadditions were predicted to be mutually orthogonal. This was validated experimentally and used for highly selective fluorescence labeling of two proteins simultaneously

Report of Acoustic Test on PSLV IS.1/2L Structure

The results of acoustic conducted on PSLV IS.1/2L at Acoustic Test Facility are briefly given. It contains test set up, Instrumentation details and tables of spectral response

Numerical Simulation of Flow Past a Prolate Spheroid

The prolate spheroid is geometrically very simple but the flow characteristics are complex which are dominated by transition and three-dimensional separation phenomenon having both primary and secondary vortex. The transition phenomenon in this flow is triggered due to the stream wise Tollmien-Schlichting (T-S) wake instability and cross flow instability, which makes the prediction more challenging due to lack of effective predictive tool to model transition. The present work aims at simulating numerically the three-dimensional turbulent flow past 6:1 prolate spheroid at Re=4.2 x 106 using RANS approach for which experimental data are available in literature. These simulations have been carried out using parallel version of the in-house multiblock structured incompressible flow solution code 3D-PURLES (3D Pressure based Unsteady Reynolds Average Navier Stokes and LES solver)

Studies on the potency of oral polio vaccine using RD cell line and evaluation of growth using different serum concentration and volume of media

Oral polio vaccine (OPV) proved to be superior in administration eliminating the need of sterile syringes and making the vaccine more suitable for mass vaccination campaigns. Poliovirus is heatsensitive in nature, and thus OPV is stored at low temperature (frozen). The growth medium containing varying concentration of serum such as 6, 8, 10, 12, 14% were prepared. 10 ml of the above mentionedgrowth media containing different concentration of serum were added to different culture bottles. The culture flasks containing different volumes of growth medium with 10% serum concentration such as 8, 9, 10, 11 and 12 ml were added to a series of culture flasks. All the culture flasks were inoculated with the RD cells (10,000 cells/culture flask) and kept at 37°C. The most favoured serum concentration and volume for the growth of RD cells was found and used for testing the potency of vaccine. Vaccinesfrom two manufacturers were kept at three different temperatures, 2-8 ± 0.5°C (refrigerator), 26 ± 0.5°C and 37 ± 0.5°C (Incubator). Cytopathic viruses were titrated by the determination of a tissue culture infectious dose50 (TCID50), vaccine dilutions were seeded in replicate onto cells in multiwell plates (usually 96 wells). After a suitable incubation period, wells were examined microscopically and scored as infected or not infected. The potency of vaccines was tested using the Karber’s Formula

EVALUATION OF PREHYPERTENSION, HYPERTENSION AND ITS ASSOCIATED FACTORS AMONG I YEAR MEDICAL STUDENTS

  Background and Objectives: Pre-hypertension is associated with increased progression to hypertension and cardiovascular risk. The objective of this study is to evaluate the prevalence of pre-hypertension and hypertension, as well as the associated factors among Ist year medical students who are from a similar socioeconomic status, dietary habits and lifestyle.Materials and Methods: This was a cross-sectional study conducted among 137 Ist year medical students using digital blood pressure (BP) monitor on three different occasions and the average was taken and also a questionnaire was obtained to assess the associated factors and other demographic details. Data were analyzed with the use of SPSS software and results were demonstrated using descriptive tables where Chi-square test and one-way analysis was used.Result: The prevalence of elevated BP (pre-hypertension and hypertension) as per JNC 7 criteria, among the medical students was 46.7%. There was a significant association of pre-hypertension with the individual risk factor like the family history of diabetes mellitus, diet and stress.Conclusion: The study shows a significant proportion of individuals with elevated BP at a younger age, associated with risk factors such as family history of diabetes, diet and stress. Elevated BP increases the risk for the development of hypertension during adolescence. Hypertension being a disease of iceberg goes unnoticed leading to chronic disease, therefore identification at the earliest can curb the disease.Keywords: Pre-hypertension, Adolescents, Prevalence, Hypertension

Transcriptome analysis of stem wood of Nothapodytes nimmoniana (Graham) Mabb. identifies genes associated with biosynthesis of camptothecin, an anti-carcinogenic molecule

Camptothecin (CPT), a monoterpene indole alkaloid, is a potent inhibitor of DNA topoisomerase I and has applications in treating ovarian, small lung and refractory ovarian cancers. Stem wood tissue of Nothapodytes nimmoniana (Graham) Mabb. (family Icacinaceae) is one of the richest sources of CPT. Since there is no genomic or transcriptome data available for the species, the present work sequenced and analysed transcriptome of stem wood tissue on an Illumina platform. From a total of 77,55,978 reads, 9,187 transcripts were assembled with an average length of 255 bp. Functional annotation and categorization of these assembled transcripts unraveled the transcriptome architecture and also a total of 13 genes associated with CPT biosynthetic pathway were identified in the stem wood tissue. Four genes of the pathway were cloned to full length by RACE to validate the transcriptome data. Expression analysis of 13 genes associated with CPT biosynthetic pathway in 11 different tissues vis-à-vis CPT content analysis suggested an important role of NnPG10H, NnPSLS and NnPSTR genes in the biosynthesis of CPT. These results indicated that CPT might be synthesized in the leaves and then perhaps exported to stem wood tissue for storage

Mutations in protocadherin 15 and cadherin 23 affect tip links and mechanotransduction in mammalian sensory hair cells

Immunocytochemical studies have shown that protocadherin-15 (PCDH15) and cadherin-23 (CDH23) are associated with tip links, structures thought to gate the mechanotransducer channels of hair cells in the sensory epithelia of the inner ear. The present report describes functional and structural analyses of hair cells from Pcdh15av3J (av3J), Pcdh15av6J (av6J) and Cdh23v2J (v2J) mice. The av3J and v2J mice carry point mutations that are predicted to introduce premature stop codons in the transcripts for Pcdh15 and Cdh23, respectively, and av6J mice have an in-frame deletion predicted to remove most of the 9th cadherin ectodomain from PCDH15. Severe disruption of hair-bundle morphology is observed throughout the early-postnatal cochlea in av3J/av3J and v2J/v2J mice. In contrast, only mild-to-moderate bundle disruption is evident in the av6J/av6J mice. Hair cells from av3J/av3J mice are unaffected by aminoglycosides and fail to load with [3H]-gentamicin or FM1-43, compounds that permeate the hair cell's mechanotransducer channels. In contrast, hair cells from av6J/av6J mice load with both FM1-43 and [3H]-gentamicin, and are aminoglycoside sensitive. Transducer currents can be recorded from hair cells of all three mutants but are reduced in amplitude in all mutants and have abnormal directional sensitivity in the av3J/av3J and v2J/v2J mutants. Scanning electron microscopy of early postnatal cochlear hair cells reveals tip-link like links in av6J/av6J mice, substantially reduced numbers of links in the av3J/av3J mice and virtually none in the v2J/v2J mice. Analysis of mature vestibular hair bundles reveals an absence of tip links in the av3J/av3J and v2J/v2J mice and a reduction in av6J/av6J mice. These results therefore provide genetic evidence consistent with PCDH15 and CDH23 being part of the tip-link complex and necessary for normal mechanotransduction

Of cattle, sand flies and men : a systematic review of risk factor analyses for South Asian visceral leishmaniasis and implications for elimination

Background: Studies performed over the past decade have identified fairly consistent epidemiological patterns of risk factors for visceral leishmaniasis (VL) in the Indian subcontinent. Methods and Principal Findings: To inform the current regional VL elimination effort and identify key gaps in knowledge, we performed a systematic review of the literature, with a special emphasis on data regarding the role of cattle because primary risk factor studies have yielded apparently contradictory results. Because humans form the sole infection reservoir, clustering of kala-azar cases is a prominent epidemiological feature, both at the household level and on a larger scale. Subclinical infection also tends to show clustering around kala-azar cases. Within villages, areas become saturated over a period of several years; kala-azar incidence then decreases while neighboring areas see increases. More recently, post kalaazar dermal leishmaniasis (PKDL) cases have followed kala-azar peaks. Mud walls, palpable dampness in houses, and peridomestic vegetation may increase infection risk through enhanced density and prolonged survival of the sand fly vector. Bed net use, sleeping on a cot and indoor residual spraying are generally associated with decreased risk. Poor micronutrient status increases the risk of progression to kala-azar. The presence of cattle is associated with increased risk in some studies and decreased risk in others, reflecting the complexity of the effect of bovines on sand fly abundance, aggregation, feeding behavior and leishmanial infection rates. Poverty is an overarching theme, interacting with individual risk factors on multiple levels. Conclusions: Carefully designed demonstration projects, taking into account the complex web of interconnected risk factors, are needed to provide direct proof of principle for elimination and to identify the most effective maintenance activities to prevent a rapid resurgence when interventions are scaled back. More effective, short-course treatment regimens for PKDL are urgently needed to enable the elimination initiative to succeed

Analysis of BAC-end sequences (BESs) and development of BES-SSR markers for genetic mapping and hybrid purity assessment in pigeonpea (Cajanus spp.)

<p>Abstract</p> <p>Background</p> <p>Pigeonpea [<it>Cajanus cajan </it>(L.) Millsp.] is an important legume crop of rainfed agriculture. Despite of concerted research efforts directed to pigeonpea improvement, stagnated productivity of pigeonpea during last several decades may be accounted to prevalence of various biotic and abiotic constraints and the situation is exacerbated by availability of inadequate genomic resources to undertake any molecular breeding programme for accelerated crop improvement. With the objective of enhancing genomic resources for pigeonpea, this study reports for the first time, large scale development of SSR markers from BAC-end sequences and their subsequent use for genetic mapping and hybridity testing in pigeonpea.</p> <p>Results</p> <p>A set of 88,860 BAC (bacterial artificial chromosome)-end sequences (BESs) were generated after constructing two BAC libraries by using <it>Hin</it>dIII (34,560 clones) and <it>Bam</it>HI (34,560 clones) restriction enzymes. Clustering based on sequence identity of BESs yielded a set of >52K non-redundant sequences, comprising 35 Mbp or >4% of the pigeonpea genome. These sequences were analyzed to develop annotation lists and subdivide the BESs into genome fractions (e.g., genes, retroelements, transpons and non-annotated sequences). Parallel analysis of BESs for microsatellites or simple sequence repeats (SSRs) identified 18,149 SSRs, from which a set of 6,212 SSRs were selected for further analysis. A total of 3,072 novel SSR primer pairs were synthesized and tested for length polymorphism on a set of 22 parental genotypes of 13 mapping populations segregating for traits of interest. In total, we identified 842 polymorphic SSR markers that will have utility in pigeonpea improvement. Based on these markers, the <it>first </it>SSR-based genetic map comprising of 239 loci was developed for this previously uncharacterized genome. Utility of developed SSR markers was also demonstrated by identifying a set of 42 markers each for two hybrids (ICPH 2671 and ICPH 2438) for genetic purity assessment in commercial hybrid breeding programme.</p> <p>Conclusion</p> <p>In summary, while BAC libraries and BESs should be useful for genomics studies, BES-SSR markers, and the genetic map should be very useful for linking the genetic map with a future physical map as well as for molecular breeding in pigeonpea.</p