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With a view to elucidating the hormonal control of decidualization in rhesus monkey, we studied the effects of CDRI-85/287, a potent anti-oestrogen, on endometrial steroid receptors in vivo and in vitro. Compound 85/287 was administered (i.m.) on days 8, 9 and 10 of steroid treatment cycle at a dose of 15 mg/monkey. Deciduoma was induced on day 16. Histological examination of endometrial tissue on days 24 and 30 of the cycle showed an apparent inhibition in uterine epithelial and subepithelial decidual cell plaque formation and a decrease in leukocytic infiltration into the stroma in anti-oestrogen-treated animals. As observed on day 24, a significant decrease in progesterone receptors (PR) (nuclear + cytosolic) was observed in the 85/287-treated groups, whereas oestrogen receptor (ER) content remained unaltered. On day 30 total ER as well as total PR content was markedly reduced in treated animals. In-vitro results clearly demonstrated a competitive antagonism of 85/287 at the ER level only. The results are discussed in relation to the histological changes and modulation of steroid receptors, thereby suggesting the decidualization inhibitory activity of anti-oestrogen molecule 85/287 in primate species. [ABSTRACT FROM PUBLISHER]

Objective: To systematically review the relationship between gingival biotype (GT) and malocclusion., Methods: The review followed PRISMA standards of quality for systematic reviews and meta-analyses reporting with PROSPERO registration number CRD42020126543. The systematic database search included MEDLINE, Scopus, Embase, PsychINFO, CINAHL, and other key journals; the article search was performed until February 2020. Cochrane's risk of bias in non-randomized studies-of interventions (ROBINS-I) was used to grade the methodological quality of the included studies., Results: The systematic search identified 105 studies, six studies satisfied the inclusion criteria for eligibility. The study participants ranged from 26 to 200 (total n=812), with a mean of 135. Study participants were aged between 14 and 32 years. Five studies were graded as the moderate risk of bias and one study as low risk of bias. Two studies showed thin GT among individuals with severe crowding compared to mild crowding. Three studies showed a thin GT with a narrow zone of the keratinized gingival width compared to a thick GT. No relationship was found between GT and Angle's classification of malocclusion., Conclusion: No relationship was observed between Angle's classification of malocclusion and GT. Thin GT was prevalent among individuals with pro-inclination of incisors. Keratinized gingival width was narrow among individuals with thin GT.

Ethanolic extract of Ferula assafoetida and chloroform fraction of Melia azedarach, both devoid of estrogenic activity, were examined for their pregnancy interceptive property. Treatment of rats from days 1 to 7 of pregnancy with either of the plant extracts resulted in pregnancy failure in about 65-85% of the animals. The possible role of energy metabolism in the antifertility action was investigated by measuring changes in activities of the key enzymes of carbohydrate metabolism in uterus on day 7 of pregnancy. It was observed that on the day 7 of pregnancy, one key enzyme of glycolytic pathway (phosphofructokinase) was significantly reduced in the uteri of treated rats as compared to controls. Hexosemonophosphate pathway also appeared to be sensitive to treatment with the plant extracts and showed an inhibitory effect on the enzyme activities of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase. Oxidative energy metabolism through tricarboxylic acid cycle, which is considered to be the main source of energy to the uterus at this stage, was maximally affected by the treatment with several enzymes showing significant inhibition. The two plant materials appeared to interrupt the latter metabolic pathway more significantly. It is thus concluded that plants lacking phytoestrogens may intercept pregnancy by their ability to disrupt energy metabolism in rat uterus during implantation, especially the oxidative pathway.

Some suitably substituted acrylophenones, quinolines and dithiocarbamate were synthesized as new generation, non-detergent spermicides and were studied for their mechanism of action in comparison with various known spermicides belonging to several different classes of chemical compound. Nonoxynol-9, benzalkonium chloride, Sapindus saponins, verapamil, emetine and tartaric acid were used as reference molecules to study the effect of new spermicides on human sperm motility parameters (using computer-assisted semen analyzer), plasma membrane integrity, lipid peroxidation and defense system against reactive oxygen species (ROS). Results have indicated that sperm plasma membrane remains the primary site of action of most of the spermicides, though the effect may be predominantly on the physiological integrity rather than the structural integrity in case of the new compounds. Lipid peroxidation may play an important role in disrupting sperm membrane physiology that may or may not be accompanied with a detrimental effect on the defense system of the human spermatozoa against the ROS.

Daily sperm production (DSP) rate was estimated in adult male rhesus and bonnet monkeys to evaluate seasonal changes in the gametogenic activity of the testes. Three monkeys of each species were castrated during breeding and non-breeding seasons and DSP rate was estimated by enumerating the homogenization-resistant spermatid nuclei of steps 13 and 14. Results indicated a significant reduction in the DSP rate per testis during the non-breeding season in two species, along with a marked decline in the testis weight. However, the gametogenic capacity of seminiferous tubules did not appear to be markedly affected during non-breeding season, as the DSP rate per gram parenchyma of testis was only marginally reduced. The seasonal changes in DSP were much more pronounced in the rhesus than in the bonnet monkey. The feasibility of circanual rhythm in DSP of sub-human primates to form a baseline for the study of reproductive function in male is discussed.

CDRI 84/35, a potent nonsteroidal antispermatogenic agent, causes total sterility in rats by directly acting on germ cells while having no effect on Sertoli/Leydig cells. This study was conducted to evaluate the effect of the compound on gametogenic activity of testes and to identify stages of spermatogenesis that were affected. Adult male rats administered either compound 84/35 at minimum effective dose or estradiol (5 micrograms) or water only were killed on days 22, 41, and 64 of the treatment period to evaluate the effect on spermatid, spermatocyte, and spermatogonial stages, respectively. Daily sperm production (DSP) was measured employing a homogenization technique. Results showed a decline in testis weight and DSP with a drastic reduction (approximately 95%) in DSP in 84/35-treated rats on day 41 of the treatment period. Estradiol was more potent in reducing the testis weight; however, 84/35 had an edge over estradiol in reducing the DSP. After withdrawal of treatment for 120 days, a phenomenal recovery (> 90%) in DSP per gram parenchyma was noted in 84/35-treated animals. Results indicate a direct effect of estradiol on spermatogonia, whereas 84/35 seems to affect the spermatocyte stage.

To assess the lipid metabolising potential of testicular germ cells undergoing meiosis, spermatocytes and spermatids were isolated from adult rat testis and purified by centrifugal elutriation followed by density gradient centrifugation. Seven key enzymes of lipid metabolism (namely beta-hydroxybutyrate dehydrogenase, carnitine acetyl transferase, ATP citrate lyase, hydroxyacyl-CoA dehydrogenase, glycerol 3-phosphate dehydrogenase, acetyl-CoA carboxylase and long chain acyl-CoA synthetase) were assayed in cell homogenates. The results indicated that germ cells possess the key enzymes for de novo synthesis and oxidation of fatty acids. The significant increase in activities of anabolic enzymes and decrease in activities of catabolic enzymes in post-meiotic germ cells indicated a shift in lipid metabolism towards fatty acid synthesis during meiosis. Long chain acyl-CoA synthetase activity was not detected in the two cell types. The study indicates a major reorganization of fatty acid turnover during meiosis with equilibrium shifting in favour of synthesis.

Studies with 85/287, a potent nonsteroidal antiestrogen/antiimplantation agent were taken up. In this paper we report alterations in uterine morphometric measurements and estrogen/progesterone receptor levels under the influence of this compound. Results showed 32% decline in stromal absolute volume density on day 5 post-coitum (p.c.) only, whereas eosinophilic leucocyte number decreased both on days 3 and 5 p.c. (41%) in treated rat uterus. Epithelial mitotic activity showed complete cessation both in control and treated rats on day 5 p.c. Under the influence of the compound both cytosolic and nuclear estrogen receptor (ERc and ERn) levels decreased on day 3 p.c., but on day 5 p.c. ERn increased significantly. A significant increase was however noticed in progesterone nuclear receptors (PRn) on day 5 p.c. On the whole our studies showed overall significant changes in uterine morphometric measurement/estrogen and progesterone receptor levels during the preimplantation period in rat under the influence of compound 85/287, causing asynchrony of events and thus failure of implantation.

CDRI compound 85/287 a potent estrogen antagonist and antiimplantation agent in rat was studied to elucidate its mechanism of action. In ovariectomized rats 85/287 treatment antagonized estrogen stimulated uterine volume density, eosinophil leucocyte infiltration, stromal mitotic cell number and peroxidase activity. In parallel experiments in pregnant rats, uterine peroxidase activity also decreased significantly as compared to controls on day 5 post-coitum. The results show that 85/287 exerts its antiimplantation activity by inhibition of responses to estradiol action.

Recent studies have indicated that estrogen has a stimulatory influence on the male reproductive tract. Evidence includes the presence of measurable levels of estrogen in the circulation, retention of exogenous estrogen, and presence of estrogen receptors in the male accessory sex organs during prepubertal life. In the present study, estrogen antagonists (CDRI-85/287 and centchroman) have been used to examine this concept by antagonising estrogen action at critical stages in the life in rat. Centchroman or 85/287 administration to 14 day old rats for 7 days did not alter gonadal or accessory organ weight. In contrast, in 21 day old castrated rats, treatment with either compound from day 28-35 of life stimulated an increase in all organ weights. When administered to normal rats during the critical phase of transition, i.e. days 30-60 of life, both testis and accessory organs showed an increase in weight. In contrast castrated rats treated with estrogen alone or in combination with 85/287 from days 37-45 of life and sacrificed on day 46 did not show any change, but 85/287 per se markedly reduced the weight of accessory organs. In adult castrated rats, the potency of DHT as a promoter of growth was potentiated by estradiol. Compound 85/287 negated the estradiol-induced increase. Glycerylphosphorylcholine (GPC) and sialic acid levels showed about 100% increase, with both high and low doses of 85/287 (treated from 30-60 days of life), However, centchroman (CDRI-67/20) was less potent in this regard. The effect of estrogen antagonists in relation to epididymal physiology during different phases of life in the rat is discussed.

Marker enzymes of Sertoli and germ cells were estimated to study the mechanism of action of antispermatogenic compound CDRI 84/35 in adult male rat testis. Animals were killed after 22, 41, and 64 days of treatment with antispermatogenic dose of CDRI 84/35 in order to evaluate the effect of the compound on spermatid, spermatocyte, and spermatogonial stages, respectively. Studies were also extended to a recovery period of 90 days. Results indicate a direction action of the compound on germ cells, with no apparent effect on Sertoli cells. Studies also show a massive depletion of postmeiotic germ cells after the treatment, with some damage to premeiotic germ cells as well. Reversibility of the compound was partial, with the marker enzymes of pre- and postmeiotic germ cells not being restored to control levels after withdrawal of treatment.

Variations in specific activities of the marker enzymes of Sertoli and germ cells during breeding (November-December) and non-breeding (May-June) seasons were investigated in rhesus and bonnet monkeys maintained under laboratory conditions. The marker enzymes selected for testicular cells were-Sertoli cells: beta-glucuronidase, gamma-glutamyl transpeptidase; pre-meiotic germ cells: glucose 6-phosphate dehydrogenase, malate dehydrogenase, alpha-glycerophosphate dehydrogenase; mature germ cells: LDH-X, sorbitol dehydrogenase. Results have indicated significant seasonal variation in marker enzymes only in rhesus testis. Marker enzymes of Sertoli cell increased while those of germ cell decreased significantly during non-breeding season. Marker enzymes of mature germ cells were affected much more drastically than those of the pre-meiotic germ cells.

The effect of androgen and estrogen antagonists on estrogen induced responses in the epididymis of rat was studied. Estradiol benzoate administered to male rates on day 5 of life increased the epididymal weight, absolute volume density of fibromuscular stroma and its eosinophilic leucocyte numbers. Testosterone administration (day 5 life) alone did not have any stimulatory effect on the epididymis as an organ or its peroxidase activity on days 15 or 20 of life. On the other hand, testosterone/85/287 negated estradiol induced increase in the absolute volume density, eosinophilic leucocyte accumulation and peroxidase activity. Tamoxifen (Tam) with inherent estrogenic activity acted both as an agonist and an antagonist. Results of present studies support the contention that nonsteroidal antiestrogens (CDRI-85/287 and Tam) can modulate estradiol induced epididymal responses during the postnatal period of male rat.

The activities and androgenic regulation of seven lysosomal enzymes viz. acid phosphatase, N-acetyl hexosaminidase, alpha-mannosidase, beta-glucuronidase, DNase II, RNase II and phospholipase A was established in caput, corpus and cauda segments of monkey epididymis. Estimation of enzyme activities in the the epididymis of control, castrated and castrated-androgen replaced monkeys revealed that all the enzymes except RNase II showed higher activity in caput and corpus as compared to cauda. The enzymes were reduced markedly after castration and on subsequent androgen replacement there was a significant stimulation of the repressed activities, but the control levels were not restored. RNase II showed highest activity in cauda which was further elevated after castration. The possible role of these enzymes in sperm maturation and disposal is discussed.

The activities of nine enzymes of the TCA cycle were estimated in the initial segment, caput, corpus and cauda segments of epididymis and vas deferens of adult rhesus monkey and expressed as units per mg DNA. These enzymes were also estimated in epididymal segments and vas deferens of castrated and castrated-androgen replaced monkeys as well. Results indicated higher activities of most of the enzymes in vas deferens as compared to epididymal segments. All the enzymes showed marked reduction in epididymis and vas deferens after castration, the effect being much more pronounced in the epididymis, than in the vas. Androgen replacement in castrated monkeys stimulated most of the enzymes markedly in epididymis and in the vas deferens as compared to their castrated values. The response of cauda and vas deferens to exogenous androgen treatment was however moderate, as compared to the other epididymal segments. The studies indicate that energy metabolism in the epididymis (as well as in the vas deferens) is strictly androgen dependent and the energy charge of these target organs is likely to fall appreciably after castration, which may in turn affect many energy dependent processes of these organs (e.g. absorption, secretion of specific substances etc.) which have been considered important for sperm maturation and survival.

In an endeavour to develop potent anti-implantation agents, a new antiestrogen, CDRI 85/287 (2-(4-2-piperidinoethoxy)phenyl-3-phenyl(2H)benzo(b)pyran), virtually devoid of agonistic activity, was identified. The present study deals with anti-implantation and estrogen agonistic-antagonistic activities of four structural analogues of 85/287. Results show that none of the compounds induced vaginal cornification, even at doses as high as 2.5 mg/kg. Compounds having p-hydroxyphenyl group at position-3 or hydroxy group at position-7 showed better estrogen receptor affinity (6.6 and 25.3% E2) as well as antiestogenic activity. When 3-p-hydroxyphenyl was replaced by 3-p-methoxyphenyl, a marked reduction in the receptor affinity was observed. However, this compound was relatively more potent as an anti-implantation agent, possibly due to its conversion to hydroxylated metabolite in vivo. The provision of aminoethoxy side chain at para-position and a shift in the piperidinoethoxy side chain from position-4 to position-2 in these molecules resulted in a decrease in estrogenicity and increase in antagonistic property. Results are discussed with regard to molecular configuration, relative binding affinity of these compounds to their biological profile.

The effect of compound CDRI-85/287, a pure, nonsteroidal antiestrogen, on implantation-associated changes in rat uterus were studied. Results provide a clear correlation between the antideciduogenic action of 85/287 (0.05 mg/kg in days 1-5 post-coitum or 2.5 mg/kg on day 1 post-coitum) and the time of its administration in relation to the secretion of prenidatory luteal phase estrogen. The antiestrogenic nature of the compound is further highlighted by inhibition of estradiol-induced increase in vascular permeability. In the present study, differences in the pattern of biochemical maturation of the pregnant, pseudopregnant and 85/287-treated rat uterus have also been illustrated. As compared to the pregnant rat uterus, absence of (the blastocyst and) decidualizing tissue in the pseudopregnant rat uterus accounts for the low uterine weight, protein, RNA, phospholipids and alkaline phosphatase at the time of implantation. Post-coital treatment with 85/287 (2.5 mg/kg, oral) inhibited increase in these parameters at the time of implantation. Glycogen levels which were lowered in the pregnant rat uterus on days 5 and 6, remain unaltered in the pseudopregnant and 85/287-treated rat uteri, suggesting nonutilization of this energy substrate. These findings provide sufficient evidence that the antiimplantation activity of 85/287 is due to its antiestrogenic property.

The androgenic regulation of the eleven enzymes of glycolytic pathway and two key enzymes of HMP pathway was studied in the initial segment, caput, corpus and cauda regions of the epididymis and in the vas deferens of rhesus monkey. The specific activities of enzymes were expressed as units of activity per mg DNA. Results in control animals indicate a marked difference in energy metabolism of epididymis and vas deferens. However, the epididymal duct itself did not show much regional variation in enzyme activities along its length. All the enzymes of the two pathways studied showed significant reduction after castration, the regulatory enzymes being affected more severely. On treatment with exogenous dihydrotestosterone, most of these enzymes showed stimulation in castrated monkeys, but none of them could be restored to normal level. The stimulation of these enzymes on treatment with exogenous dihydrotestosterone varied along the epididymal duct itself being maximum in the initial segment and minimum in the cauda region. The changes in the vas deferens were less marked as compared to the epididymis following castration and androgen replacement.

Ability of compound CDRI-85/287, a new nonsteroidal antiestrogen with negligible inherent estrogenicity, to inhibit uptake of 3H-estradiol (3H-E2) by the immature rat uterus in vivo was investigated. Different doses of 85/287 were administered either intraperitoneally 30 min before 3H-E2 or orally 1 and 6 hr before 3H-E2. A dose dependent inhibition in 3H-E2 uptake was observed after administration of the compound by either route and was 69% at 50 micrograms/rat ip dose and 80% at 2.5 mg/kg po dose. In in vitro competitive binding assay, however, the compound showed poor affinity (RBA 0.42% of estradiol-17 beta) for cytosolic estrogen receptors. Considering the potent anti-estrogenic as well as anti-implantation efficacy of the compound, its action in vivo appears to be mediated via its active metabolite(s).

Duration of antiestrogenic and antiimplantation action of CDRI-85/287, (2-(4-(2-N-piperidino)ethoxy phenyl)-3-phenyl(2H)benzo(2)pyran), was studied in rat. Pretreatment of ovariectomized immature rats with this compound caused translocation of cytoplasmic estrogen receptor (ER) to the nucleus and a marked depletion of cytoplasmic ER pool resulting in a nonresponsive state of the uterus to subsequent estrogen administration until day 4. While in rats pretreated with estradiol, increased cytoplasmic ER level made the uterus responsive to a second injection of estrogen. In the delayed implantation model, 85/287 pretreated rats were given estrone on days 4, 5 or 6 post-antiestrogen treatment. No implantations were observed after estrone administration on day 4, but were present when estrone was given on days 5 or 6. Summation of these results suggests the duration of action of 85/287 to be 3-4 days in rat.

The importance of exocrine secretions of testis in the regulation of energy metabolism of the epididymis and vas deferens was examined in rhesus monkeys by performing efferentiectomy. At autopsy the epididymis was divided into initial segment, caput, corpus and cauda portions to make an account of regional differences, if any. Eleven enzymes of glycolysis, two key enzymes of HMP pathway and seven enzymes of TCA cycle were assayed in the epididymal segments and vas deferens of control (intact) and experimental (efferentiectomised for 90 days) monkeys. The results indicate that while anaerobic energy metabolism (glycolysis and HMP pathway) is sensitive to efferentiectomy chiefly in the proximal regions of epididymis, the oxidative pathway (TCA cycle) is dependent on testicular exocrine secretions throughout the length of epididymis, as well as in the vas deferens. Since all androgen-sensitive enzymes do not regress after efferentiectomy, it is suggested that unidentified exocrine factors of testis may have role in regulating energy metabolism in the epididymis and vas deferens.

The ethanolic extract of A. aureum and its fractions were evaluated for postovulatory antifertility activity in female rats. The water soluble fraction of ethanolic (95%) extract prevented (100%) pregnancy when administered to female rats on days 1-7 postcoitum. This fraction was found devoid of both estrogenic and antiestrogenic activities.

Postcoital antifertility efficacy, estrogenic and antiestrogenic activities of compound 85/287 were determined by the subcutaneous route in rats. It was 100% effective in preventing implantation at 0.5 mg/kg dose when administered within 24 h of mating and at 0.05 mg/kg in the days 1-5 post-coitum regimen. In the immature rat bioassay, it exhibited mild uterotrophic effect at the contraceptive dose but when administered along with estradiol (E2), it caused almost complete inhibition of uterine weight gain and vaginal cornification at the 2 mg/kg dose. E2 administration to immature rats (0.1 microgram, s.c., 3 days) caused 3-5 fold increase in the nuclear as well as cytoplasmic estradiol receptor (ER) content as compared to controls. In contrast, 85/287 (0.5 mg/kg and 2 mg/kg; s.c.), only translocated the ER to the nuclear compartment resulting in a depletion of cytoplasmic ER levels. Concurrent administration of 85/287 and E2 inhibited E2-induced increase in cytoplasmic ER. It is suggested that compound 85/287 exerts its antiestrogenic and antiimplantation action by interfering with the formation of E2-receptor complexes in the uterus.

Compound CDRI-85/287: 2-[4-(2-N-piperidinoethoxy) phenyl]-3-phenyl (2H) benzo (b) pyran has been identified as a potent antiimplantation agent in rat. A single oral dose (2.5 mg/kg body weight) of the compound administered on days 1, 2 or 3 of pregnancy or multiple dosing (0.05 mg/kg daily) on days 5-7 postcoitum effectively prevented pregnancy. When administered on days 5-7 postcoitum, it failed to interrupt pregnancy even at 20 mg/kg dose. The compound is a potent antiestrogen, with very weak uterotrophic activity; it does not induce vaginal cornification in immature ovariectomised rat. Also, it is devoid of progestational, antiprogestational, androgenic, antiandrogenic and antigonadotrophic activities. The results suggest that the compound exerts its antiimplantation acivity in rat by virtue of its antiestrogenic activity [corrected].

The possible physiological role of estrogen in the regulation of energy metabolism of epididymis and vas deferens of rhesus monkey was investigated. A few selected key enzymes of glycolysis (hexokinase, phosphofructokinase and pyruvate kinase) and TCA cycle (succinate dehydrogenase and malate dehydrogenase) were measured in these two organs of (a) castrated estrogen treated, (b) castrated estrogen + dihydrotestosterone (DHT) treated animals and compared with those in castrated and castrated + DHT treated animals. Results reveal that DHT stimulated the activities of all these enzymes whereas estrogen failed to stimulate any of the enzymes in castrated animals. However, estrogen in combination with DHT caused a marked stimulation of the enzymes and the response of the epididymis and vas deferens to combination treatment was significantly more than that caused by DHT alone. The results suggest that circulating estrogen in male has a physiological role and acts synergistically with androgen in regulating accessory sex organ function.

The effect of Anandron, a nonsteroidal antiandrogen, on the reproductive system and fertility of adult male rats was studied. Administered at a daily oral dose of 5 mg and 10 mg (per rat) for 30 days, it caused a significant increase in the plasma testosterone levels. Spermatogenic arrest in about 20 to 50% of the tubules in 9 out of 16 rats and stimulation of Leydig cells was observed in rats treated with the higher dose. Although a reduction occurred in accessory sex organ (epididymis, seminal vesicles, SV; ventral prostate, VP; dorsal prostate, DP and coagulating gland, CG) weight, no parallel reduction was evident in the secretory indices of the epididymis (glycerylphosphorylcholine and sialic acid), VP (alkaline phosphatase) and CG (fructose). However, there was a reduction in the total content per organ of these constituents. Females mated with treated males showed postimplantation loss indicating an adverse effect of Anandron during spermiogenesis. The results suggest that the peripheral antiandrogenic potency of Anandron in intact animals is insufficient to completely neutralize the elevated levels of androgens. A microdose (1 microgram) of estradiol efficiently neutralized the central stimulatory effect of Anadron and potentiated its antiandrogenic action. The potential use of such a combination for 'Fertility Regulation' in male is discussed.

Ultrastructural changes in the epididymal epithelium and the fate of accumulating spermatozoa were examined in the vasectomized rhesus monkey (Macaca mulatta). Accumulation of spermatozoa resulted in an increase in the diameter of the tubule and its lumen. Ultrastructure of principal cells revealed that they continue to perform both secretory and absorptive functions after vasectomy. The rough endoplasmic reticulum, Golgi complex, and mitochondria were well developed in the principal cell. Bulging of the apical portion of principal cells and membrane-bound structures in the lumen suggests an increase in apocrine secretion. An increase in the number of vesicles, vacuoles, and multivesicular bodies in the principal cells indicates an increased absorptive activity. Increased absorptive function was also evident in the apical cells. Macrophages with sperm remnants were seen in the lumen, and occasionally in the connective tissue. The principal or only mechanism of sperm disposal after vasectomy appeared to be intraluminal endocytosis by macrophages.

Anandron (5,5-dimethyl-(4-nitro-3(trifluoromethyl)phenyl)-2,4-imidazolidine-dione ), a nonsteroidal antiandrogen was administered orally to growing rats from day 31 through day 60 of age to study its effect on the testis and genital tract. At a daily dose of 5 mg per rat, it caused a significant reduction in the weights of testis, epididymis, seminal vesicles, ventral prostate and dorsal prostate. In more than 50 percent of the animals, it caused arrest of spermatogenesis at spermatid stage. Though there was a reduction in the epididymal content of glycerylphosphorylcholine and sialic acid, their concentration was not altered much. The results are discussed in relation to the effects of anandron on the genital organs of adult male rat.

The zinc concentration in the epididymis (caput, corpus and cauda regions), vas deferens and caudal lobe of prostate of adult rhesus monkeys was determined by atomic absorption spectrophotometry. Zinc content (microgram/g wet weight) was found to be maximum in the prostate (709 micrograms) followed by epididymis and vas deferens. The three segments of the epididymis did not differ from one another in their zinc content (165-177 micrograms). On a protein basis maximum concentration of zinc was present in the nuclear fraction followed by microsomal, cytosolic and mitochondrial fractions in that order. Ligation of testicular efferent ducts or castration 90 days prior to autopsy caused a marked reduction in zinc concentration in different sub-cellular fractions of the organs examined; castration was relatively more effective in this regard. The importance of androgen and other testicular products in controlling zinc content and the possible physiological role of zinc in the male genital tract are discussed.

Centchroman [dl-3,4-trans-2,2-dimethyl-3-phenyl-4-[p-(beta-pyrrolidinoethoxy)phenyl] - 7-methoxychroman hydrochloride], an antifertility agent under clinical evaluation, has been resolved into its optical enantiomers. The cytosol estrogen receptor binding affinity and estrogenic, antiestrogenic and antiimplantation activities of the two enantiomers have been determined. The enantiomers display a 7-fold difference in receptor affinity, and a corresponding difference in stimulation of the uterine growth and antiimplantation activity was observed in rats.

Zinc level in maturing spermatozoa collected from caput, corpus, and cauda regions of the epididymis and vas deferens of rhesus monkey was measured. Spermatozoa collected from the caput epididymis have the highest concentration of zinc (36 micrograms/10(8) spermatozoa). There was a marked reduction in the zinc content of corpus spermatozoa and a further decrease in the spermatozoa of cauda epididymis. Thus the zinc content of spermatozoa gradually decreases as they migrate from the caput to cauda epididymis. There was a significant increase in the zinc content of vasal spermatozoa as compared to those collected from the cauda region of epididymis.

This study was undertaken with a view to investigate the possible mechanism(s) of antifertility action of gossypol acetate in rats and hamsters. Adult male rats were treated by gavage with 30 mg/kg/day of gossypol for 7 weeks and adult male hamsters were treated similarly with 20 mg/kg/day gossypol for 8 weeks. The treatment caused a marked reduction in the weights of testis and epididymis. Histological examination of the testis in the two species revealed presence of seminiferous tubules showing varying degrees of damage along with a large number of normal tubules. Exfoliation of germ cells and spermatogenic arrest at spermatid stage was a common feature. Leydig cells presented normal morphological features. Though there was a reduction in the diameter of epididymal tubules, the epithelium did not show any morphological alterations. Examination of vasal flushings revealed marked reduction in sperm population and consisted of decapitated and immotile spermatozoa. Gossypol caused a significant reduction in the levels of total protein, RNA and DNA, and a marginal decrease in glycogen content in the testis. This was accompanied by a reduction in the activities of SDH and MDH. Except for LDH activity which showed a marked rise, there was no effect on glycolytic enzymes in the testis. The concentrations of glycerylphosphorylcholine and sialic acid were reduced in the cauda epididymis. The antifertility effects of gossypol appear to be due to its action both on testis as well as on epididymis.

The effect of flutamide, a non-steroidal antiandrogen on the growth and secretory function of the epididymis of rat during transition from prepubertal age (35 days old) to puberty (50 days old) was investigated. The results showed that the antiandrogen interfered with the growth of all the genital organs accompanied by a marked reduction in the secretory function of the epididymis as revealed by lowered levels of sialic acid and glycerylphosphorylcholine. These findings are discussed in relation to our previous observations in adult rats.

The regional fine structure of the epithelium lining the epididymis of the rhesus monkey has been investigated. Tall, prismatic principal cells constituted the major part of the epithelium. Their basal plasma lemma showed presence of caveolae and the luminal surface was studded with stereocilis. Presence of numerous desmosomes and tight junctions between adjacent principal cells suggested the existence of blood epididymis barrier. Ultrastructural evidence is presented in support of high lysosomal turnover, absorptive and secretory functions in these cells. Apocrine secretion was evident only in the principal cells of initial segment. Of the two types of basal cell, dark and pale, the latter revealed presence of lipofuscin pigment suggesting their scavenger role in the epithelium.

Administration of a low dose of estradiol (0.25 or 2.5 microgram/animal) to immature rats caused a pulsatile receptor translocation, resulting in a single nuclear receptor peak (1-3 hr) and maintenance of the uterine growth until 24 hr. At a higher dose (10.0 microgram/rat), maintaining the circulatory estradiol levels for a longer duration, a biphasic nuclear translocation occurred. The usual profile of nuclear receptor binding until 12 hr was followed by a second phase of receptor translocation, resulting in an additional nuclear receptor peak at 24 hr. The uterus continued to grow until 72 hr, reaching five times its original wet weight. The duration of receptor interaction and the magnitude of uterine stimulation would, therefore, appear to be largely dependent upon the period of bioavailability of estradiol. However, there are additional intracellular regulatory mechanisms not fully understood as yet, which seem to modulate the cytosol-nuclear receptor dynamics, thus influencing the extent of uterine stimulation.

In a study of the effect of the substituent on the receptor binding affinity (RBA), estrogenicity, and antiimplantation (AI) activity in trans-3,4-diarylchromans, it has been found that demethylation of trans-2, 2-dimethyl-3-phenyl-4-[p-(beta-pyrrolidinoethoxy)phenyl]-7-methoxychroman (centchroman, 1) to the corresponding 7-hydroxy compound (7) results in a 20-fold increase in RBA (112%) without any appreciable change in AI activity. On the other hand, absence of the pyrrolidinoethyl group from the 4-phenyl residue (6) leads to a drop in both RBA and AI activity. A chain length of two to three carbon atoms and a pyrrolidino ring appear to be necessary for activity in these compounds. It has been found that while the trans isomers with the tertiary aminoalkoxy side chain in the para position of the 4-phenyl radical were the most active, in the corresponding cis-chromans and chromenes, analogues with this chain in the meta position were most active; the ortho substituted compounds of all these series were inactive. In 3-phenyl-substituted compounds, the trans isomer carrying the p-hydroxy substituent (33) was found to be the most active; the corresponding pyrrolidinoethyl ether (13) showed a lower order of activity. The implication of these observations on the mapping of the different subsites on the receptor has been discussed.

The structural and functional integrity of the epididymis, the acquisition of fertilizing ability by spermatozoa and their viability within the epididymis are androgen dependent phenomena. Although the precise mechanism by which sperm maturation and viability in the epididymis are brought about by androgen are not clearly understood, it is generally held that specific epididymal secretions produced under the influence of androgen affect these events. Though the spermatozoa appear to remain viable in a low androgen environment, sperm maturation requires a relatively high androgen environment. Against this background the potentiality of antiandrogens as extragonadal antifertility agents has been discussed. Studies with steroidal and nonsteroidal antiandrogens have revealed that in adult animals the secretory activity of the epididymis, as evidenced by the level of glycerylphosphorylcholine, either remains unaffected or is stimulated under their influence. These studies have further indicated that the extragonadal antifertility action of antiandrogens will depend upon their ability to (1) lower the testicular androgen synthesis and/or androgen binding protein, which possibly serves as a carrier of androgen from the testis to epididymis; (2) to lower local androgen synthesis as a result of reduced levels of circulating androgen, and (3) to inhibit 5 alpha-reduction of testosterone to dihydrotestosterone and/or to inhibit androgen binding to receptors. Success in the rational development of new antifertility agents for male which will act by controlling epididymal function will depend upon a clear understanding of the factors that regulate epididymal secretion and the role of epididymal secretions in sperm maturation and survival.

Effect of three antiandrogens: cyproterone acetate (5 mg/day, sc), flutamide (5 mg/day, sc) and STS-557 (5 mg/day, po) and an estrogen, estradiol dipropionate (5 micrograms/day, sc) on some key enzymes of carbohydrate metabolism was investigated in adult rat epididymis and ventral prostate. Antiandrogens were administered for 21 days and estrogen for 14 days. All of them caused a significant decrease in the weight of epididymis, seminal vesicles and ventral prostate. A significant decrease in the specific activities of enzymes (hexokinase, phosphofructokinase, aldolase, glyceraldehyde phosphate dehydrogenase, pyruvate kinase, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase) occurred only in the organs of estrogen treated rats; activities of some of the enzymes were lowered also in the prostate of STS-557 treated rats. Flutamide and cyproterone acetate were ineffective in this regard. The possible factors responsible for the ineffectiveness of synthetic antiandrogens in influencing epididymal metabolism are discussed.

Synthesis and antiimplantation activity of variously substituted 2,2-dialkyl-3,4-diphenylchromenes and 3,4-cis- and trans-chromans derived from them are described. Pregnancy-inhibiting activity in rats was exhibited by a number of these compounds, which was particularly marked in the case of 3,4-trans-3-phenyl-4-p-(beta-pyrrolidinoethoxy)-phenyl-7-methoxychroman (32), the corresponding 2,2-dimethyl analog 34, and 3-phenyl-4-p-(beta-pyrrolidinoethoxy)phenyl-7-methoxychromene (26). The structure-activity relationship of these compounds is discussed.

With a view to impart selective uptake of estrogen by the target tissues of rat, liposomes in which (6,7-3H) oestradiol-17 beta constituted a part of lipid bilayer were used as carriers of the hormone. The distribution and retention of the radioactivity was determined in blood plasma, uterus, liver, kidney, spleen and leg muscle of ovariectomized rat at different time intervals up to 72 hr following a single intravenous injection of free oestradiol (1.61 muCi) or an equivalent amount of liposomal-oestradiol. When free oestradiol was administered, uterus showed peak amount of radioactivity between 30 min to 2 hr and remained high up to 6 hr. In the other tissues examined, maximum amount of radioactivity was seen at 15 min followed by a marked fall at 30 min and also at other subsequent intervals. The pattern of uptake and retention of radioactivity after administration of liposomal-oestradiol was not much different from that of free oestradiol between 1 and 6 hr. A moderate increase in the amount of radioactivity in the nongenital tissues at 24 hr was the only difference noticed with liposomal-oestradiol. It is concluded that targeting of oestradiol preferentially to the uterus could not be achieved through liposomal delivery system.

The effect of flutamide (Sch 13521; 4'-nitro-3'-trifluoromethylisobutyranilide), a nonsteroidal antiandrogen, on male rat genital organs was studied. Administered at a dose of 25 mg/kg body weight daily for 30 days, flutamide caused a significant increase in the weight of the testis but had no effect on spermatogenesis and Leydig cell morphology. The secretory activity of the epididymis, as evidenced by the level of glycerylphosphoryl-choline and sialic acid, either remained unaffected or was stimulated. There was a significant decrease in seminal vesicle and ventral prostate weight and in the fructose content of the coagulating gland. The anti-androgen at the dose used did not affect the fertility of the rats. The significance of these findings is viewed in relation to the hypothesis of a differential threshold requirement of androgen for the epididymis and the accessory sex glands. The potentiality of antiandrogens as extragonadal antifertility agents in the male is discussed.

A new cervical dilator, Isaptent, was prepared from granulated Plantago ovata (Isapgol) seed husk. It was evaluated in a multicentric clinical trial for dilatation of the cervix in subjects opting for medical termination of pregnancy. The trial covered 804 women in over 21 centres in different parts of the country. The cases were between 15 to 45 years of age, 0 to 10 parity with a gestation period of 8 to 24 weeks. A single tent was used in 750 subjects and satisfactory dilatation was achieved in 94% of the cases. The cervical dilatation bore no relationship to age, parity and gestation period of the subjects. The tent provided self-lubrication, caused no apparent damage to the cervix and the vaginal flora remained unchanged in the randomly selected subjects in whom bacteriologic studies were performed. The outcome of the clinical trial and advantages of Isaptent over the other procedures used for cervical dilatation are discussed.

The in vitro metabolism of (3H)-androstenedione in the epididymis and vas deferens of intact and castrated rats was investigated and the metabolites formed were identified by radio gas chromatography. Incubation of slices of caput epididymidis for 2 hr at 34 degrees C metabolised 90% androstenedione. Similar incubations of tissue samples from cauda epididymidis and vas deferens metabolized 60 and 25% of androstenedione respectively. The major metabolites formed in the epididymis were androstanedione (caput: 48%; cauda: 33%) and androsterone (caput: 35%; cauda: 13%). These metabolites appeared in much less concentration in the incubations with vas deferens (about 8% each). In general, conversion to testosterone and dihydrotesterone was low in all the three organs examined. Castration did not significantly alter the metabolic pattern in the caput epididymidis and vas deferens but promoted the formation of androsterone (38%) in the cauda epididymidis. The conversion of androstenedione, a weak androgen to testosterone, dihydrotestosterone and 3 alpha/3 beta-diols in the epididymidis and vas deferens of castrated rats may be of physiological significance. In addition, androsterone appears to be an important androgenic metabolite in the epididymis.