Expression and Detection of Recombinant Sheep Prion Protein (RecShPrP^C) Using Human Neuroblastoma Cells and Western Blotting

The objective of this study was to over express the sheep normal prion protein (RecShPrPc) in SK-N-SH cells using a mammalian expression vector with C-terminal 6XHis-tag for large scale purification by Nickel-Nitrilotriacetate column chromatography. The full length prion open reading frame was PCR amplified and the complete coding sequence (PrP25-234) including the signal peptide from both C and N terminal ends of the protein was subcloned into pcDNA6.0 plasmid. The orientation and integrity of the cloned PCR product in the mammalian expression vector was confirmed by DNA sequencing. The prion gene was transfected using Lipofectamine2000 reagent and successfully over expressed in SK-N-SH cells. The cells were lysed and the over expressed prion protein was detected using Western Blot. Production of RecShPrPC using this method would ensure the availability of large amounts of this protein for further downstream applications. In vitro production of the full length PrPC and its detection is important in further studies aimed at deciphering the biochemistry and structure of prion proteins.Department of Animal Scienc

Study on the reproductive behavior among women of rural areas of Pondicherry

Background: The fertility rate in India is declining and it is necessary to know the factors responsible for such decline in different states. It was decided to study reproductive behavior of women contributing to decline in fertility in Pondicherry. The aim and objective of this study is known the reproductive behavior of women in the rural areas on Pondicherry.Methods: Sample of 300 married women aged between 30 and 60 years were selected randomly from village belonging to Katerikuppam PHC and the data collected using the pre tested semi open ended questionnaires by interviewing the subjects at their doorsteps during September to November 2014.Results: The mean age of the subjects was 41.5 ± 9.5 years. There were 793 total pregnancies and live births were 701 and abortions accounted for ten percent and still births were 9 in number. The mean number of pregnancies and live births were 2.6 ± 1.1 and 2.3 ± 1.0 per women respectively. The mean age at menarche, marriage and first pregnancy were 14.3 ± 1.4, 19.6 ± 3.1 and 21.1 ± 3.1 years respectively. The mean number of pregnancies were declined from currently older age to the lower age of the subjects and found to be statistically significant.Conclusions: The mean age at marriage and first pregnancy is increasing and the differences in the mean number of pregnancies and live births are showing declining trend.

Expression and Localization of CLC Chloride Transport Proteins in the Avian Retina

Members of the ubiquitously expressed CLC protein family of chloride channels and transporters play important roles in regulating cellular chloride and pH. The CLCs that function as Cl−/H+ antiporters, ClCs 3–7, are essential in particular for the acidification of endosomal compartments and protein degradation. These proteins are broadly expressed in the nervous system, and mutations that disrupt their expression are responsible for several human genetic diseases. Furthermore, knock-out of ClC3 and ClC7 in the mouse result in the degeneration of the hippocampus and the retina. Despite this evidence of their importance in retinal function, the expression patterns of different CLC transporters in different retinal cell types are as yet undescribed. Previous work in our lab has shown that in chicken amacrine cells, internal Cl− can be dynamic. To determine whether CLCs have the potential to participate, we used PCR and immunohistochemical techniques to examine CLC transporter expression in the chicken retina. We observed a high level of variation in the retinal expression levels and patterns among the different CLC proteins examined. These findings, which represent the first systematic investigation of CLC transporter expression in the retina, support diverse functions for the different CLCs in this tissue

Synthesis and Surface-Active Properties of Sodium <i>N</i>‑Acylphenylalanines and Their Cytotoxicity

Sodium <i>N</i>-acylphenylalanines (NaNAPhe) were synthesized using a mixture of fatty acids obtained from coconut, palm, karanja, <i>Sterculia fetida</i>, and high oleic sunflower oils via the Schotten–Baumann reaction in 60–78% yield to see the influence of the hydrophobic group of fatty acyl group functionality with headgroup phenylalanine on their surface-active properties. The products were characterized by chromatographic (thin-layer, column, and gas) and spectral (IR, NMR, and mass) techniques. The synthesized products were evaluated for their surface-active properties such as surface tension, wetting power, foaming characteristics, emulsion stability, calcium tolerance, critical micelle concentration (CMC), and thermodynamic properties. The results showed that all of the products exhibited superior surface-active properties like CMC (0.018–0.00041 mmol/L), calcium tolerance (26.5–65.8 ppm), and emulsion stability (262–844 s) compared to the reference sodium lauryl sulfate. All of NaNAPhe except sodium <i>N</i>-coconut fatty acylphenylalanines exhibited promising cytotoxicity against human cancer cell lines. These new vegetable oil based surfactants have potential in personal care, pharmaceutical, and industrial applications

Total synthesis of (−)-Sachalinol A and evaluation of its cytotoxicity

695-700(-)-Sachalinol A, a group of monoterpenoids, oxygenated derivatives of rosinidol, has been synthesized employing a simple, eight step procedure. The compound demonstrates excellent cytotoxicity against MDA-MB-231 derived from human breast adenocarcinoma cells (ATCC No. HTB-26) and HeLa derived from human cervical cancer cells (ATCC No. CCL-2). Good activity is observed against A549 derived from human alveolar adenocarcinoma epithelial cells (ATCC No. CCL-185) and Neuro2a derived from mouse neuroblastoma cells (ATCC No. CCL-131). Moderate activity is also observed against MCF-7 derived from human breast adenocarcinoma cells (ATCC No. HTB-22)

Role of Hormones and the Potential Impact of Multiple Stresses on Infertility

Infertility has a remarkable global impact affecting approximately about 48 million couples worldwide. One of the most contended concerns in reproductive biology is the prospective influence of psychological stress on pregnancy rates. Individuals struggling to conceive face a stupendous amount of emotional turbulence and have a greater risk for psychological vulnerability. Both stress and infertility reinforce the impact of each other; hence, there exists a reciprocal relationship. Stress may be the major contributor to subsequent infertility. Infertility treatments may enhance stress levels as well as reduce treatment outcomes. The biological mechanisms that interlink stress and infertility are the outcome of the hormonal actions at the brain level, where they stimulate or suppress the hypothalamic-pituitary-adrenal axis (HPA) and have a potential influence on the secretion of the respective hormone by the reproductive organs and the pregnancy outcomes. Sex hormones play an essential role in reproductive biology as well as in general physiology where they generate the cycle and provide a potential environment for pregnancy. This article reviews the impact of stress on reproductive rates and the implications of sex hormones on infertility. Additionally, it suggests strategies to overcome the stress conditions and the scenarios that may lead to stress

Induction of circulating T follicular helper cells and regulatory T cells correlating with HIV-1 gp120 variable loop antibodies by a subtype C prophylactic vaccine tested in a Phase I trial in India.

A Phase I HIV-1 vaccine trial sponsored by the International AIDS Vaccine Initiative (IAVI) was conducted in India in 2009 to test a subtype C prophylactic vaccine in a prime-boost regimen comprising of a DNA prime (ADVAX) and MVA (TBC-M4) boost. The trial demonstrated that the regimen was safe and well tolerated and resulted in enhancement of HIV-specific immune responses. Preliminary observations on vaccine-induced immune responses were limited to analysis of neutralizing antibodies and IFN-γ ELISPOT response. The present study involves a more detailed analysis of the nature of the vaccine-induced humoral immune response using specimens that were archived from the volunteers at the time of the trial. Interestingly, we found vaccine induced production of V1/V2 and V3 region-specific antibodies in a significant proportion of vaccinees. Variable region antibody levels correlated directly with the frequency of circulating T follicular helper cells (Tfh) and regulatory T cells (Treg). Our findings provide encouraging evidence to demonstrate the immunogenicity of the tested vaccine. Better insights into vaccine-induced immune responses can aid in informing future design of a successfulHIV-1 vaccine