PubMed Health⌕ Search

Biomedical subjects

A H Bandivdekar

Publications and source records attributed to A H Bandivdekar.

At least 19 recordsLinked to original sources
PubMed2006

HOXBES2: a novel epididymal HOXB2 homeoprotein and its domain-specific association with spermatozoa.

The sperm from the testis acquires complete fertilizing ability and forward progressive motility following its transit through the epididymis. Acquisition of these characteristics results from the modification of the sperm proteome following interactions with epididymal secretions. In our attempts to identify epididymis-specific sperm plasma membrane proteins, a partial 2.83-kb clone was identified by immunoscreening a monkey epididymal cDNA library with an agglutinating monoclonal antibody raised against washed human spermatozoa. The sequence of the 2.83-kb clone exhibited homology to the region between 1 and 1097 bp of the homeobox gene, Hoxb2. This sequence was found to be species conserved, as revealed by RT-PCR analysis. To obtain a full-length clone of the sequence, 5' RACE-PCR (rapid amplification of cDNA ends PCR) was carried out using rat epididymal RNA as the template. It resulted in a full-length 1.657-kb cDNA encoding a 32.9-kDa putative protein. The protein designated HOXBES2 exhibited homology to the conserved 61-amino acid homeodomain region of the HOXB2 homeoprotein. However, characteristic differences were noted in its amino and carboxyl termini compared with HOXB2. A putative 30-kDa protein was detected in the tissue extracts from adult rat epididymis and caudal spermatozoa, and a 37-kDa protein was detected in the rat embryo when probed with a polyclonal antibody against HOXB2 protein. Multiple tissue Western blot and immunohistochemical analysis further indicated its expression in the cytoplasm of the principal and basal epithelial cells, with maximal expression in the distal epididymal segments. Northern blot analysis detected a single approximately 2.5-kb transcript from the adult epididymis. Indirect immunofluorescence localized the protein to the acrosome, midpiece, and equatorial segments of rat caudal and ejaculated human and monkey spermatozoa, respectively. In conclusion, we have identified and characterized a novel epididymal homeoprotein different from HOXB2 protein and hereafter referred to as HOXBES2, (HOXB2 homeodomain containing epididymis-specific sperm protein) with a probable role in fertilization.

Acrosome↗
PubMed2006

Identification and characterization of oviductal glycoprotein-binding protein partner on gametes: epitopic similarity to non-muscle myosin IIA, MYH 9.

The mammalian estrogen induced oviductal glycoprotein (OGP) has been known to associate with capacitated sperm, oocytes and developing embryos. This study aimed to identify the putative binding partner of OGP on gametes using N-terminal peptide of bonnet monkey (Macaca radiata) OGP, Nmon, as bait. A protein(s) of molecular size approximately 54 kDa was detected by far-western blot analysis of detergent solubilized human sperm proteins. MALDI-TOF mass spectra analysis of approximately 54 kDa tryptic peptides gave a significant hit to non-muscle myosin heavy chain. Biochemical characterization of approximately 54 kDa was done with antibodies specific to non-muscle myosin IIA, MYH9. The approximately 54 kDa protein, possible breakdown product of MYH9, immunoreacted with MYH9 antibody in western blot analysis. OGP binding to approximately 54 kDa could also be demonstrated in far-western blot analysis of detergent solubilized human sperm proteins and nuclear matrix intermediate filament (NM-IF) preparations from human sperm and mouse oocytes. Far-western blot analysis of MYH9 enriched by immunoprecipitation identified the native approximately 220 kDa protein as OGP-binding partner. The identical and characteristic immunogold localization pattern of Nmon and MYH9 on sperm NM-IF preparation substantiated these findings. The results suggest that OGP binds to both gametes through its interaction with MYH9 through the non-glycosylated N-terminal conserved region of OGP, spanning the residues 11-137.

Animals↗
PubMed2005

Effect of Lomodex-MgSO(4) in the prevention of reperfusion injury following unilateral testicular torsion: an experimental study in rats.

Fertility in patients treated for unilateral testicular torsion has been shown to be significantly reduced in all the reported series to date, implying that the present-day treatment requires further refinement in the form of adjunct pharmacotherapeutic intervention (Lomodex and MgSO(4)) in addition to scrotal exploration. Prepubertal Holtzman strain rats (35 days old) were used for our study. Two sets were formed with six groups of rats in each set. Rats were treated as follows: group 1, sham-operated group; group 2, torsion (4 h); group 3, torsion + detorsion (1 h); group 4, torsion + ATP-MgCl(2) + detorsion; group 5, torsion + Lomodex-MgSO(4) + detorsion; group 6, torsion + normal saline + detorsion. Whereas the first set of animals was sacrificed immediately at the end of experiment, animals in set 2 were sacrificed 8 weeks after the end of the experiment to look for the development of antisperm antibodies. Parameters studied were thiobarbituric acid reductase (TBAR) assay, histology of testicular tissue, and sperm agglutination test. Student's t-test was used for significance. With detorsion (149.95+/-30.68) there was a significant rise in the TBAR values (P<0.05) compared with torsion (57.39+/-14.47). Treatment with both Lomodex-MgSO(4) (40.74+/-6.39) and ATP-MgCl(2) (48.30+/-18.35) yielded TBAR levels comparable to those in the sham group (31.35+/-11.96). Similar injury was also seen on the contralateral testis, with detorsion (114.28+/-10.68) much more detrimental than torsion (40.59+/-15.02) and rescue seen following treatment with Lomodex-MgSO(4) (27.55+/-8.64) as well as ATP-MgCl(2) (38.61+/-12.23). Regarding th histology, with detorsion there was evidence of severe distortion of tubules, with almost all the tubules showing maturation arrest and a few tubules completely devoid of any germinal cells. Treatment with Lomodex-MgSO(4) as well as ATP-MgCl(2) showed preservation of tubular morphology. Our study failed to document the presence of agglutinating antibodies (antisperm antibodies) in any of the groups. Unilateral testicular torsion has bilateral effects and is a form of ischemia-reperfusion injury. Treatment of torsion by detorsion alone does not prevent testicular damage. The results of the present study show that administration of Lomodex + MgSO(4) prior to detorsion results in prolonged testicular salvage with a potential of subsequent improvement in semen quality and fertility and reduction in long-term morbidity. The presence of agglutinating antibodies could not be detected in the present study.

Adenosine Triphosphate↗
PubMed2004

Studies with Synthetic Peptides of 80 kDa Human Sperm Antigen (80 kDa HSA).

PROBLEM: The 80 kDa human sperm antigen (HSA) is a sperm-specific and conserved antigen, capable of inducing immunological infertility. Partial N-terminal amino acid sequences of 80 kDa HSA (Peptide NT) and its peptides obtained by digestion with endoproteinase Lys-C (peptides 1-4) and endoproteinase Glu-C (peptides 5-6) did not show any sequence homology with reported known proteins deposited in the Gen-Bank. These sequenced peptides were synthesized and conjugated to key hole limpet haemocyanin (KLH) and evaluated for its antifertility effects. The present communication describes the characterization of these peptides and their antibodies. METHOD OF STUDY: Peptides NT, 1, 2, 3 and 4 were synthesized and conjugated to KLH. Antibodies to KLH conjugated peptides were raised in rabbits by active immunization and the antibody titer was determined by enzyme-linked immunosorbent assay (ELISA) using sperm extract coated wells. The binding specificity of the synthetic peptides or purified 80 kDa HSA to their antibodies was assessed in the presence of various doses of respective synthetic peptides or 80 kDa HSA. The binding specificity was further confirmed by Western blot analysis. Antipeptide antibodies were also checked for sperm agglutinating activity, in-vitro. RESULTS: Active immunization of rabbits elicited significant antibody titers against the synthetic peptides, except for peptide 3. Antipeptide antibodies specifically recognized the native protein in an ELISA and induced in-vitro agglutination of human, rat and monkey sperm. In addition, Western blot analysis showed that these antipeptide antibodies specifically bind to the 80 kDa HSA band of the sperm extract. CONCLUSION: Synthetic peptides of 80 kDa HSA are immunogenic and antibodies raised against these peptides recognize the native protein detected by ELISA, Western blot analysis. In addition, they possess sperm agglutinating activity. These findings suggest that they are promising candidates in the development of immunocontraceptives.

Agglutination Tests↗
PubMed2001

Characterization of 80 kDa human sperm antigen responsible for immunoinfertility.

PROBLEM: An 80 kDa human sperm antigen (80 kDa HSA) has been identified by western blot technique using serum of an immunoinfertile woman as a probe. The 80 kDa HSA has been subsequently purified from sperm extract and investigated for antifertility effects. METHOD OF STUDY: The purified 80 kDa HSA was used to immunize adult male and female rats. Rabbit anti 80 kDa HSA antibodies were used for immunofluorescent and immunohistochemical staining to demonstrate the presence of 80 kDa HSA on the sperm and to investigate its tissue distribution. The N-terminal sequence of native 80 kDa HSA and the peptides obtained by its endoproteinase Lys-C was determined. RESULTS: Active immunization of male and female rats with 80 kDa HSA caused infertility in all the immunized animals. Immunofluorescent staining showed its localization on the head region of the human and rat spermatozoa. While immunohistochemical studies showed its localization in the testes and epididymis but not in other somatic tissues. Partial amino-acid sequence analysis showed no sequence homology with any of the known protein in the database. CONCLUSIONS: 80 kDa HSA is a promising candidate antigen for immunocontraception as it is characterized and found to cause infertility upon active immunization. specific to spermatozoa and is conserved.

Amino Acid Sequence↗
PubMed1992

Antifertility effects in rats actively immunized with 80 kDa human semen glycoprotein.

A 80 kDa human sperm antigen has been identified using the serum of an infertile woman having circulating antisperm antibodies. The antigen was then purified to homogeneity by gel permeation chromatography using HPLC (protein PAK-125 column) system and on FPLC (superose-12 column) system. The antigen was found to be a glycoprotein. The antigen was mainly localized in the postacrosomal region of the human sperm, while it was localized in the head region of the rat sperm as demonstrated by immunofluorescent staining. The presence of this antigen was also demonstrated in the human prostate and endometrium and in the rat testis; epididymis and the prostate by immunocytochemical staining. The purified protein upon active immunization in female rats caused infertility in 100 percent animals. While in male rats it caused infertility in 90 percent animals. On morphometric analysis of testicular tissue it was observed that there was no significant change in spermatogonia and spermatocytes, but significant decrease in spermatids and sperm number as well as daily sperm production in the immunized male rats. The epididymal spermatozoa were markedly reduced in number and were largely found to be agglutinated. The results suggest that 80 kDa human sperm antigen appears to be a suitable candidate for immunocontraception both in male and female.

Animals↗
PubMed1991

Antifertility effects of human sperm antigen in female rats.

The sperm antigens responsible for inducing infertility were identified by Western blot technique using sera from an infertile woman with circulating antisperm antibodies. The 80 Kda was prepared from human sperm by extraction with 0.05% sodium deoxycholate in 0.01 M Tris-HCl buffer, pH 8.4 and fractionation with ammonium sulphate. The supernatant after 40% saturation ammonium sulphate extraction was separated by gelpermeation chromatography, using HPLC (Protein PAK 125 column) and FPLC (superose 12 column) systems. The homogeneity of the protein was established by SDS-PAGE and its molecular size was estimated to be 80 Kda and its isoelectric point was 4.5. The purified protein upon active immunization in female rats caused infertility in 100 percent animals. The data suggest that 80 Kda human sperm antigen has the potential for use as a contraceptive vaccine.

Animals↗
PubMed1990

Modulation of lipid peroxidation in human spermatozoa and human prostate by prostatic inhibin.

Loss of sperm motility as a result of the production of hydrogen peroxide by lipid peroxidation is regulated by as yet unidentified prostatic factor(s). Inhibinlike peptide of prostatic origin isolated from human seminal plasma, with a molecular size of about 10,400 daltons, was studied for its effect on ascorbate-induced lipid peroxidation in human spermatozoa. Dose-related suppression of lipid peroxidation was observed at dose levels of 0.25, 0.5, and 1.0 micrograms. The data suggest that inhibinlike peptide could be one of the factors involved in the regulation of lipid peroxidation and thereby of sperm motility. Inhibinlike peptide also exhibited local action in both normal and benign hyperplastic human prostate tissue by enhancing the rate of lipid peroxidation. These findings have implications in the pathophysiology of the prostate.

Dose-Response Relationship, Drug↗
PubMed1989

Modulation of prolactin release by a 25 kD peptide of human seminal plasma.

Prolactin-regulating factor (PRF) of molecular size of 25 kD has been isolated from human seminal plasma. This 25 kD factor inhibits circulating PRL levels in intact adult male rats to the extent of about 48% at dose level of 10 micrograms. Furthermore, in vitro incubation of pituitary demonstrated that PRF inhibits the release of PRL in the medium. PRF also interferes with the binding of I125 PRL to its receptors in liver, prostate, and spermatozoa. However, I125 PRF itself does not bind to these receptors. PRF seems to modulate PRL release as well as its binding to receptors. A sensitive, specific RIA was developed for PRF. Using the RIA, levels of PRF in seminal plasma were measured. PRF levels were low in vasectomized subjects as compared with controls. A negative correlation was noted with seminal plasma PRF levels and sperm count.

Animals↗
PubMed1989

Suppression of lipid peroxidation in human spermatozoa by prostatic inhibin.

Loss of sperm motility owing to the production of hydrogen peroxide by lipid peroxidation is regulated by yet unidentified prostatic factor(s). Inhibinlike peptide (HSPI) of prostatic origin isolated from human seminal plasma and having a molecular weight of about 10,400 daltons was studied for its effect on ascorbate-induced lipid peroxidation in human spermatozoa. Dose-related suppression of lipid peroxidation occurred at a dose level of 0.25, 0.5, and 1.0 micrograms. HSPI may be one of the factors involved in the regulation of lipid peroxidation and therefore sperm motility.

Humans↗
PubMed1989

Prostatic inhibin-like peptide quantified in urine of prostatic cancer patients by enzyme-linked immunosorbent assay.

This is a highly specific enzyme-linked immunosorbent assay (ELISA) for measuring prostatic inhibin-like peptide (PIP) in urine, in which we use penicillinase (EC 3.5.2.6) conjugated with PIP and, as solid phase, a polystyrene microtiter plate. We used this ELISA to measure PIP in 24-h urine specimens from men with prostatic cancer (PCa) and from age-matched controls. For prostatic cancer patients the mean +/- SEM urinary PIP of 36.1 +/- 5 micrograms/24 h was significantly (P less than 0.001) lower than the mean of 127.1 +/- 9 micrograms/24 h for the age-matched controls. PIP values for 30 samples measured by both ELISA and RIA correlated well (r = 0.985). We could detect as little as 1.56 ng of PIP in a sample. Analytical recovery of added PIP ranged from 91% to 104%. Mean CVs were 8.9% within-assay and 12.7% between-assay. We believe that this ELISA will be useful in assessing the status of PIP in men with normal and diseased prostates and in examining the function of the hypothalamus-pituitary-prostate axis.

Aged↗
PubMed1988

Evidence for mutually antagonistic actions of thyroid releasing hormone and inhibin at pituitary-gonadal-prostate-spermatozoal axis.

In-vivo as well as in-vitro studies carried out demonstrate for the first time the interaction between thyroid releasing hormone (TRH) and inhibin at the pituitary, testes, prostate and spermatozoa levels. At the pituitary level both peptides act as antagonists to each other and modify the release of pituitary hormones. Further, these peptides act at the prostatic level wherein they modulate ornithine decarboxylase activity as well as 5 alpha-reductase activity. At the testicular level TRH blocks inhibin biosynthesis whereas in semen, it significantly reduces the binding of spermatozoa to specific antibodies directed against inhibin. In conclusion, although TRH and inhibin are widely different in their molecular size and chemical structure, these peptides seems to act antagonistically at multiple sites.

Animals↗
PubMed1988

Hypothalamic-hypophyseal-testicular abnormalities and erectile dysfunction.

Forty-five men presenting with erectile dysfunction were evaluated through history and nocturnal penile tumescence, Doppler, and EMG studies. Fifteen were classified as having organic and 30 as having psychogenic impotence. Three men had mild hypergonadotropism with low testosterone levels. One had hyperprolactinemia. No case of hypogonadotropic hypogonadism was detected. Six patients who had psychogenic impotence had low levels of testosterone.

Adolescent↗
PubMed1987

Antibodies to human seminal plasma inhibin cause sperm agglutination and impairment of cervical mucus penetration and sperm-egg attachment.

Inhibin isolated from human seminal plasma which has 94 amino acids has been shown to be structurally similar to a sperm coating antigen of prostatic origin. Specific antibodies generated against this peptide caused agglutination of human sperm. Using FITC-labeled antibody, antigen was localized on the post-acrosomal head region of sperm. Antiserum to inhibin could also impair the penetration of human spermatozoa into cervical mucus. After 10 and 30 minutes, the depth and density of penetration as well as the motility of the sperm were inhibited. The treatment of sperm with antiserum to inhibin caused an inhibition of sperm attachment to the egg as well as inhibition of penetration.

Cervix Mucus↗
PubMed1985

Endocrine control of inhibin biosynthesis by human placenta.

In vitro synthesis of inhibin-like activity was localized in the fetal part of the human placenta. Of the various hormones, hCG stimulated inhibin synthesis while progesterone, estradiol, LHRH and prostaglandin inhibited the synthesis. Prolactin did not significantly alter the inhibin synthesis.

Chorionic Gonadotropin↗