Dietary fluted pumpkin seeds induce reversible oligospermia and androgen insufficiency in adult rats.

Some components of the human diets are believed to be promising male contraceptive agents. The present study examined the antispermatogenic efficacy, reversibility and toxicity of fluted pumpkin seed-supplemented diet (DFPS) in adult male Wistar rats. Adult rats were given DFPS at 2.5, 5 and 10% for 60 days followed by 60 days post-treatment period. The control animals received normal standard rat diet not supplemented with fluted pumpkin seeds. The sperm quality variables, testosterone and follicle-stimulating hormone (FSH), oxidative status of the testis, steroidogenic enzymes and gamma-glutamyl transferase (γ-GT) activities and the histology of the testis were determined to evaluate the anti-fertility activity of fluted pumpkin seeds. Treatment of animals with DFPS at 5% and 10% resulted in decreased serum and intra-testicular testosterone and FSH concentrations. This effect was associated with decreased activity of 17ß-hydroxysteroid dehydrogenase (17ß-HSD), increased testicular oxidative stress and poor sperm quality in the 10% diet group. After 60 days DFPS post-treatment, intra-testicular 17ß-HSD and γ-GT activities, FSH and testosterone levels recovered to control values. Furthermore, poor sperm motility, count, morphology and viability as well as severe loss of spermatogonia and other matured epithelial germ cells and Sertoli cells observed especially in the 10% DFPS-treated animals reverted to nearly control values 60 days after withdrawal of treatment. Dietary fluted pumpkin seeds may selectively act on the epithelial germ cells, possibly mediated via Sertoli cells, leading to oligospermia, oxidative damage and androgen insufficiency. The reversibility of these effects to near normal levels after withdrawal of treatment justifies further consideration of DFPS as it may be an effective and readily reversible agent that meets the required criteria of a male contraceptive agent.Abbreviations: GC-MS: gas-chromatography mass spectrophotometry; MPO: myeloperoxidase; NO: nitric oxide; DFPS: dietary fluted pumpkin seeds; DFPS (REV): DFPS post-treatment; MDA: malondialdehyde; SOD: superoxide dismutase; CAT: catalase; GSH: glutathione; 3ß-HSD: 3ß-hydroxysteroid dehydrogenase; 17ß-HSD: 17ß-hydroxysteroid dehydrogenase; NAD: nicotinamide adenine dinucleotide (oxidized); NADH: nicotinamide adenine dinucleotide (reduced); ITT: intra-testicular testosterone; FSH: follicle-stimulating hormone; FPS: fluted pumpkin seeds; NIST: National Institute Standard and Technology; Av: absolute volume; Ac: cross-sectional area; ST: seminiferous tubules; γ-GT: gamma glutamyl transferase.

Characterization of EPPIN's semenogelin I binding site: a contraceptive drug target.

Epididymal protease inhibitor (EPPIN) is found on the surface of spermatozoa and works as a central hub for a sperm surface protein complex (EPPIN protein complex [EPC]) that inhibits sperm motility on the binding of semenogelin I (SEMG1) during ejaculation. Here, we identify EPPIN's amino acids involved in the interactions within the EPC and demonstrate that EPPIN's sequence C102-P133 contains the major binding site for SEMG1. Within the same region, the sequence F117-P133 binds the EPC-associated protein lactotransferrin (LTF). We show that residues Cys102, Tyr107, and Phe117 in the EPPIN C-terminus are required for SEMG1 binding. Additionally, residues Tyr107 and Phe117 are critically involved in the interaction between EPPIN and LTF. Our findings demonstrate that EPPIN is a key player in the protein-protein interactions within the EPC. Target identification is an important step toward the development of a novel male contraceptive, and the functionality of EPPIN's residues Cys102, Tyr107, and Phe117 offers novel opportunities for contraceptive compounds that inhibit sperm motility by targeting this region of the molecule.

Reversible antifertility effect of aqueous leaf extract of Allamanda cathartica L. in male laboratory mice.

The efficacy of oral administration of aqueous leaf extract of Allamanda cathartica (150 mg kg(-1) body weight day(-1) for 14, 28 and 42 days) in inducing infertility and changes in various male reproductive endpoints was evaluated in Parkes strain mice. The effect of the treatment on organ weight, histopathology, sperm parameters, testosterone level, haematology, serum biochemistry and on fertility indices was assessed. Histologically, testes in extract-treated mice showed nonuniform degenerative changes in the seminiferous tubules as both affected and normal tubules were observed in the same sections. The treatment also had adverse effects on motility, viability, morphology and on number of spermatozoa in the cauda epididymidis. Serum levels of testosterone, alanine aminotransferase, aspartate aminotransferase and creatinine, haematological parameters and liver and kidney histoarchitecture were, however, not affected by the treatment. Fertility of the extract-treated males was also suppressed, although the libido remained unaffected. By 56 days of treatment withdrawal, however, the above parameters recovered to control levels. Our results thus suggest that A. cathartica treatment causes reversible suppression of fertility in male mice, without causing detectable toxic effects.

Chiral separation of rac-Ornidazole and detection of the impurity of (R)-Ornidazole in (S)-Ornidazole injection and raw material.

(S)-Ornidazole is a subject of research as an antifertility agent in male animals at present. However, there seems to be no relative report on chiral separation for rac-Ornidazole, which has been used as an effective medicine for more than 30 years. In this article, the chiral separation of rac-Ornidazole on a Chiralcel OB-H column based on normal-phase high-performance liquid chromatography (NP-HPLC) is investigated and the methodology for detection of impurity of (R)-Ornidazole in (S)-Ornidazole injection and raw material is established. The novel mobile phase is utilized by mixing n-hexane, methanol and isopropyl alcohol (95:4:1, v/v/v) instead of the typical mobile phase of n-hexane and isopropyl alcohol, although the methanol, which offers a good resolution factor for the enantiomeric separation in this system, is not recommended on the Chiralcel OB-H column according to the instruction supplied by Daicel Chemical Ind., LTD (Japan).

Study on the role of a sperm membrane-associated antigen in canine fertilization.

The Mab 4B12 produced by us against capacitated boar spermatozoa was found to recognize a protein located in the acrosome portion of capacitated boar spermatozoa which is shared by different animal species, dogs included. It was shown that Mab 4B12 might affect fertilizing ability in vitro of boar spermatozoa. Using indirect immunofluorescence (IIF) test, we provide evidence here that Mab 4B12 stained the acrosome of the capacitated but not of the ejaculated and acrosome-reacted canine spermatozoa. The biological experiments using hemizona assay functional test in this study provide evidence supporting the involvement of Mab 4B12 corresponding antigen in the functional steps required for canine sperm-zona pellucida binding. These results together with the data on cell and tissue specificity of the 4B12 antigen suggest its contraceptive potential for canine fertilization.

Conhecimento e uso do condom: anticoncepçäo e prevençäo de doenças sexualmente transmissíveis / Knowledge and condom use: contraceptive method and prevention of disease of sexual transmission

Até recentemente o conhecimento sobre a incidência de uso do condom o considerava apenas como método anticoncepcional e provinha, na grande maioria das vezes, de pesquisas sobre fecundidade, conduzidas junto a mulheres unidas e na idade reprodutiva. O excluir mulheres näo unidas, o näo considerar homens, e pensar o Condom apenas como método contraceptivo, säo vieses metodológicos que explicam a reduzida taxa de uso deste preservativo em nosso meio. Procurou-se na pesquisa, cujos resultados ora se apresentam, investigar o conhecimento, uso e razöes de uso do condom junto ao universo masculino privilegiando-se, em uma primeira etapa, 3 segmentos específicos, a saber: universitários, bancários e operários, com idade entre 18 e 30 anos. Neste sentido, foi possível estimar taxas diferenciais de uso do condom e registrar as preocupaçöes que se delineam nos grupos estudados quanto a conduta sexual, frente a crescente presença da AIDS

The male antifertility activity of 6-chloro-6-deoxyglucose.

6-Chloro-6-deoxy[U-14C]glucose is not metabolised by mature boar spermatozoa nor has it any specific inhibitory action on their metabolic activity in vitro. The compound is metabolised by the male rat and the identification of two urinary metabolites as alpha-chlorohydrin and 3-chlorolactate confirmed that (S)-3-chlorolactaldehyde is produced by this species in vivo. A tissue distribution study revealed that radioactivity from 6-chloro-6-deoxy[U-14C]glucose was more concentrated in rat caudal spermatozoa than in any other of the major tissues.

Long-acting contraceptive agents: testosterone esters of unsaturated acids.

The synthesis of 13 new esters of testosterone is described, with the esterifying acids bearing acetylenic, olefinic, or polyunsaturated functions in the chain, for evaluation as long-acting androgens.

PIP: A program of the World Health Organization for developing long-acting esters of testosterone that would exhibit a more constant release rate and maintain testosterone levels in the normal range longer than testosterone enanthate found that these esters had a role in fertility, and gerontology. The synthesis of 13 new esters of testosterone is described, with the esterifying acids bearing acetylenic, olefinic or polyunsaturated functions in the chain, for evaluation as long-acting androgens. The nuclear magnetic resonance (NMR) images were recorded on a spectrometer. The samples were recorded in tubes using CDC13 as solvent. The NMR spectra were recorded with Perkin-Elmer instrument in CDC13, with tetramethylsilane as internal reference. Infrared spectra were measured on the same spectrometer. Mass spectra were also recorded. Thin-layer chromatography was performed on Merck silica gel and the spray reagent was iodine or vanillin. To a solution of testosterone the corresponding acid chloride was added yielding the pure ester after the usual work-up. E-5-methylhexa-2,4-dienoic (IXb), 5- phenylpenta-2-,4-dienoic (Xb), 5-phenyl-4-yn-pent-2-enoic (XIb), and non-4-en-6-ynoic acid (XIIb), were required for the synthesis. Esterification of testosterone with each of the first 12 unsaturated acids was performed by reaction with the corresponding acid chlorides in pyridine. Although the nona-2,3-dienoic acid ethyl ester was easily obtained, this compound could not be hydrolyzed to the acid (XIIIb). Hence, an alternative procedure was tried for the synthesis of the ester XIIIa, by reaction of bromoacetate of testosterone (XIVa) with triphenylphosphine to give the phosphorane (XVa). Reaction of this phosphorane (XVa) with 1-diazoheptan-2-one (XVI) led to the allenic ester (XIIIa).