N-acetyl-L-cysteine counteracts oxidative stress and prevents H2O2 induced germ cell apoptosis through down-regulation of caspase-9 and JNK/c-Jun.

The mechanism of H(2)O(2) induced oxidative stress leading to male germ cell apoptosis was earlier reported from our laboratory. In the present study, we investigated the mechanisms by which N-acetyl-L-cysteine (NAC, which is highly cell specific with strong antioxidant and anti-genotoxic properties), stimulated cell survival under such conditions. Co-incubation with 5 mM NAC significantly (P<0.

Enzyme linked immunosorbent assay for milk progesterone.

A direct antigen heterologous enzyme linked immunosorbent assay (ELISA) for milk progesterone has been developed using progesterone-3-O-carboxymethyloxime-bovine serum albumin (P-3-O-CMO-BSA) antiserum and 17-α-hydroxy-progesterone-3-O-carboxymethyloxime-horseradish peroxidase (17-α-OH-P-3-O-CMO-HRP) enzyme conjugate. The data of the present study reveal that the homologous assay, which employed P-3-O-CMO-HRP as the label, showed no displacement. On the contrary, replacement of P-3-O-CMO-HRP with 17-α-OH-P-3-O-CMO-HRP as the label showed significant displacement and led to the development of a sensitive and specific assay.

Development of heterologous enzyme linked immunosorbent assay for dehydroepiandrosterone in serum.

Homologous and heterologous combinations of enzyme conjugate with immunogen in steroid enzyme immunoassay (EIA) influence labeled steroid recognition by an antibody that affects the sensitivity of the assay. To develop dehydroepiandrosterone (DHEA) enzyme linked immunosorbent assay (ELISA), antibodies were generated against dehydroepiandrosterone-17-carboxymethyloxime-bovine serum albumin (DHEA-17-CMO-BSA) and dehydroepiandrosterone-7-carboxymethyloxime- bovine serum albumin (DHEA-7-CMO-BSA). Two dehydroepiandrosterone (DHEA) horse radish peroxidise (HRP) enzyme conjugates were prepared using two dehydroepiandrosterone derivatives (DHEA-17-CMO and DHEA-7-CMO).

AR versus ER (α) expression in the testis and pituitary following chronic estrogen administration in adult rat.

Modulation of the testis-pituitary axis has direct relevance to the expression of androgen and estrogen receptors. Androgen receptor (AR) and estrogen receptor (ERα) expression during hypospermatogenesis after chronic estrogen administration to rats was studied in the adult testis and pituitary utilizing immunohistochemistry, western blotting, and RT-PCR. Both organs demonstrated higher AR transcriptional activity gradually increasing from 15 days (d) to 30 d of estrogen treatment.

Altered levels of seminal nitric oxide, nitric oxide synthase, and enzymatic antioxidants and their association with sperm function in infertile subjects.

Objective: To investigate the association between specific reactive nitrogen species and sperm function among infertile subjects.

Design: Controlled trial.

Setting: Fertility clinic at the National Institute of Health and Family Welfare, New Delhi, India.

Providers' knowledge, attitude and dispensing practices of e-pills in government dispensaries of South district in delhi, India.

Background: South Delhi is one of the well developed districts in the capital with best public health care facilities. Knowledge, attitude and dispensing practices of emergency contraceptive pills (E-pills) were assessed among health care providers of government dispensaries in South Delhi.

Study Design: A descriptive epidemiological study.

Effect of chronic oestrogen administration on androgen receptor expression in reproductive organs and pituitary of adult male rat.

Following chronic (15 or 30 days) treatment with oestradiol 3-benzoate (75 microg rat(-1) day(-1) in 100 microl of olive oil) to adult rats, androgen receptor (AR) expression was analysed simultaneously in testis, epididymis, seminal vesicle, prostate and pituitary utilising three independent tools i.e. immunohistochemistry, Western blotting and RT-PCR.

Health research strengthening and operational research needs for improving child survival in India.

Health research can be utilized to improve the policies, interventions and outputs of the health systems, and ultimately the health of individuals and population. This requires systematic evaluation of evidence and its integration into national policies and programs after suitable adoption at the local level. It has been noted that there has been limited focus upon strengthening health research in India, due to weak research systems or institutional mechanisms, lack of trained human resources and enabling environment, absence of well defined priorities, perceived low quality of research, and inadequate funding.

Sperm function and seminal oxidative stress as tools to identify sperm pathologies in infertile men.

Routine semen analysis, which includes assessment of sperm motility, viability, and morphology, does not always provide complete diagnostic information as men demonstrating standard scores on these specific parameters sometimes remain infertile. In infertile men who had been subgrouped on the basis of sperm count, motility, morphology, and presence of leukocytes, an evaluation of sperm function and reactive oxygen species (ROS) was found to be effective tools to detect sperm pathologies.

Release of copper from CuT 380A co-incubated with human semen and its effect on sperm function in vitro.

Release of copper and its effect on functional integrity of human sperms in vitro were assessed following co-incubation of semen with CuT 380A. After 30 min of incubation with semen, release of copper ions from CuT 380A was found to be 9.2 to 40 times higher compared to control incubations with PBS.

Trials for development of once-a-month injectable, hormonal male contraceptive using dienogest plus testosterone undecanoate: dose standardization, efficacy and reversibility studies in rats.

Background: The study was conducted to test the potential of using dienogest (DNG) plus testosterone undecanoate (TU) in rats for development of a once-a-month injectable male hormonal contraceptive.

Study Design: Dose selection studies were initiated with administration of DNG in three different doses of 20, 30 and 40 mg/kg body weight (bw) per week plus TU 25 mg/kg bw once in every 6 weeks. Status of spermatogenesis and sperm count in epididymis was evaluated.

Development of ELISA for measurement of progesterone employing 17-alpha-OH-P-HRP as enzyme label.

The present study was aimed to develop a highly specific and sensitive Enzyme Linked Immunosorbent Assay (ELISA) to measure progesterone in human serum using a heterologous combination of immunogen and enzyme conjugate. The antiserum was raised against Progesterone-3-O-carboxymethyloxime bovine serum albumin (P-3-O-CMO-BSA) in New Zealand white rabbits. The enzyme conjugate was prepared by labeling 17-alpha-hydroxy-progesterone-3-O-carboxymethyloxime (17-alpha-OH-P-3-O-CMO) with Horseradish Peroxidase (HRP) to form 17-alpha-OH-P-3-CMO-HRP.

Pathways involved in testicular germ cell apoptosis induced by H2O2 in vitro.

H(2)O(2) induces apoptosis in variety of cells; however, the sensitivities of testicular germ cells to H(2)O(2) are not known. In the present study, H(2)O(2), at concentrations in the range 1-10 microM, was found to induce apoptosis in testicular germ cells in vitro. Following 1 h of treatment with 10 microM H(2)O(2), a 10-fold rise in the percentage of apoptotic cells was observed.

Deciphering a conformation-specific epitope of hCG-beta through Immunokinetics.

Proteins and peptides are comprised of both sequence-specific and conformation-specific epitopes. Sequence-specific epitopes are delineated by a peptide approach and other robust methods like competition assays, gene expression assays, synthetic peptide library based assays, etc. Available methods for deciphering conformation-specific epitopes are cumbersome (X-ray crystallography, etc.

Kinetic analysis of a human chorionic gonadotropin-beta epitope-paratope interaction.

Kinetics of protein-protein or ligand-ligate interaction has predominantly been studied by optical spectroscopy (particularly fluorescence) and surface plasmon resonance biosensors. Almost all such studies are based on association kinetics between ligand-ligate and suffer from certain methodological and interpretational limitations. Therefore, kinetic analyses of dissociation data of such interactions become indispensable.

A novel enzyme-linked immunosorbent assay for cortisol using a long-chain biotinylated cortisol-3-CMO derivative.

An enzyme-linked immunosorbent assay (ELISA) using streptavidin-biotin system as a bridge between antibodies bound antigen and reporter molecule (horseradish peroxidase enzyme) has been described. The cortisol antiserum was generated against cortisol-3-O-carboxylmethyl oxime-bovine serum albumin (F-3-CMO-BSA). We have prepared biotin-labelled cortisol as a primary probe and utilized streptavidin-labelled horseradish peroxidase (SA-HRP) as secondary probe to monitor the antigen-antibody interaction.

Use of biotin-streptavidin system for developing a viable, sensitive and specific antigen heterologous assay for hapten.

The present study demonstrates improvement in sensitivity and specificity of hapten assay by using antigen heterology in conjunction with low molecular weight biotin label as compared to high molecular weight horseradish peroxidase (HRP) label. For generation of antiserum, cortisol-3-O-carboxylmethyl-oxime-bovine serum albumin (F-3-CMO-BSA) was used as immunogen whereas, for the preparation of primary label, corticosterone-3-carboxymethyl oxime (B-3-CMO) was coupled with biotinylcaproylhydrazide and HRP by employing N-hydroxysuccinimide mediated carbodiimide reaction. The data of the present study revealed that the antigen heterologous assay which employed high molecular weight HRP label showed 100% cross-reaction with corticosterone.

Estimation of mortality due to AIDS--a review.

HIV/AIDS has emerged as a major public health problem since its recognition as an emerging disease a couple of decades ago. While detection of HIV/AIDS cases remains a problem, ascertainment of AIDS deaths has emerged as a bigger challenge and concern. Despite a plethora of literature focusing on the methods to estimate AIDS deaths, none seems to be fulfilling the requirements for universal acceptance.

Development of rapid and sensitive one-step direct enzyme linked immunosorbent assay for 17-alpha-OH-progesterone in serum.

Using a homologous combination of immunogen and enzyme conjugate, a highly specific and sensitive Enzyme Linked Immunosorbent Assay (ELISA) was developed to measure 17-alpha-hydroxy-progesterone (17-alpha-OH-P) in human serum. The antiserum was raised against 17-alpha-hydroxy-progesterone-3-O-carboxymethyloxime bovine serum albumin (17-alpha-OH-P-3-O-CMO-BSA) in New Zealand white rabbits. The enzyme conjugate was prepared by labeling 17-alpha-hydroxy-progesterone-3-O-carboxymethyloxime with horseradish peroxidase (HRP).

Is abnormal sperm function an indicator among couples with recurrent pregnancy loss?

Objective: To determine whether or not sperm function parameters are altered in male partners of couples with a history of idiopathic recurrent pregnancy loss (RPL).

Design: In comparison with proven fertile volunteers, sperm function parameters like hypo-osmotic swelling (HOS), acrosomal status (AS), and nuclear chromatin decondensation (NCD) were assessed in vitro from male partners of couples with a history of idiopathic RPL.

Setting: Infertility clinic and andrology laboratory at National Institute of Health and Family Welfare.

Verapamil reversibly inhibits spontaneous parthenogenetic activation in aged rat eggs cultured in vitro.

The present study was designed to investigate whether verapamil could inhibit spontaneous parthenogenetic activation in aged rat eggs cultured in vitro. Eggs collected from oviduct after 19 h post human chorionic gonadotropin (hCG) were arrested at the metaphase-II (M-II) stage and exhibited a first polar body. Culture of these aged eggs in calcium/magnesium (Ca(2+)/Mg(2+))-deficient and serum-free medium for 3 h induced exit from M-II, a morphological sign of spontaneous parthenogenetic activation in all eggs.

Antigen heterologous enzyme linked immunosorbent assay for the measurement of estrone-3-glucuronide.

We report a novel antigen heterologous enzyme linked immunosorbent assay for the direct estimation of estrone-3-glucuronide (E1-3-G) in diluted human urine. The differential behavior of antibody towards the labeled and unlabelled analyte in heterologous systems forms the basis of the present assay. Antiserum was raised in New Zealand white rabbits using estrone-3-glucuronide-bovine serum albumin (E1-3-G-BSA) as immunogen and nandrolone-17-HS (N-17-HS) coupled to horseradish peroxidase (HRP) with and without urea bridge ((N-17-HS-Urea-HRP/N-17-HS-HRP) as an enzyme labeled reagent.

Influence of hydrophobic and hydrophilic spacer-containing enzyme conjugates on functional parameters of steroid immunoassay.

Introduction of spacers in coating steroid antigen or enzyme conjugates or immunogen is known to exert an influence on the sensitivity of steroid enzyme immunoassays. We have introduced hydrophobic and hydrophilic spacers between enzyme and steroid moieties and studied their effects on functional parameters of enzyme immunoassays, using cortisol as a model steroid. Cortisol-3-O-carboxymethyloxime-bovine serum albumin (F-3-O-CMO-BSA) was used as immunogen to raise the antiserum in New Zealand white rabbits.

Development of dual-enzyme-based simultaneous immunoassay for measurement of progesterone and human chorionic gonadotropin.

The development of a simultaneous multianalyte immunoassay for the detection of progesterone and human chorionic gonadotropin (hCG) in serum is described. In this simultaneous multianalyte assay, two different enzymes, viz. horse radish peroxidase (HRP) and alkaline phosphatase (ALP), were used as markers.

Development of a rapid, sensitive, and reproducible laboratory test kit for the assessment of plasma membrane integrity of human sperm.

Objective: To develop a rapid, sensitive, and reproducible hypoosmotic swelling test kit for the assessment of plasma membrane integrity of human sperm in vitro.

Design: Prospective comparison of results with the World Health Organization (WHO) method, performed simultaneously.

Setting: Infertility center in a major city in India.