Buchholzia coriacea (wonderful kola) seeds induce male reproductive toxicity by suppressing the pituitary-gonadal axis in Wistar rats

The methanolic extract of Buchholzia coriacea seeds (MEBC) has been reported to induce male reproductive toxicity by decreasing sperm parameters and fertility index. To elucidate the possible mechanism(s), the effects of graded doses of MEBC on sex hormones and sperm profile were investigated in this study. The MEBC (e.g., 50, 200, 400, and 600 mg/kg) was administered daily (p.o.) to male Wistar rats for 6 weeks, while a concurrent control group received distilled water (vehicle). Then, the animals were sacrificed under sodium pentobarbital anaesthesia. Weights of organs were recorded, and the sperm profile was determined microscopically. Testosterone, luteinizing hormone (LH), and follicle stimulating hormone (FSH) were assayed from the obtained serum using the ELISA technique. Sperm motility was significantly reduced by MEBC (i.e., 50 and 200 mg/kg), and sperm count reduced in all treated groups in a dose-dependent manner compared with that of the control. Serum testosterone, LH, and FSH decreased in treated rats. A histopathological examination of testes showed a considerable depletion and necrosis of the epithelium of seminiferous tubules. The result suggests that Buchholzia coriacea seeds induce male reproductive toxicity by suppressing the pituitary-gonadal axis.

Pharmacological effects of Sesbania sesban: a systematic review

The cost, side effects and imitation associated with conventional drugs have driven a substantial number of global citizens to resort to complementary medicine. Although largely informal and unregulated, the practice of herbal medicine is more engrained in low and middle income than in industrialized countries. Sesbania sesban, a plant which grows generously across most parts of the world, has been a major target by most traditional health practitioners. The effects so far reported include antimicrobial, anti-fertility, anti-diabetic, anti-inflammatory among others. No study has reviewed the scholarly works published and jointly reported results. Authros systematically reviewed papers available in different databases to give a hybrid report on the pharmacological effects of Sesbania sesban. A total of three data bases were searched using key terms like: Sesbania sesban, ethno-botany, phytochemical analysis, pharmacological effects etc. A total of 860 papers were initially recovered and further subjected to abstract and title examination which filtered them down to 40 papers. The 40 papers were assessed more against a set of criteria like: in-vivo and in-vitro studies biased to pharmacological effects of the plant, studies that were less than 15 years old and studies that used experimental design. This further scrutiny reduced the number of papers to 25. Most studies reported Sesbania sesban as having great anti-microbial, anti-fungal, anti-inflammatory and anti-diabetic qualities. No study reported any adverse effect of the plant. Authors recommend a dose-effect assessment and mechanism of action of the plant extracts especially with regard to the antimicrobial, anti-inflammatory and anti-fertility qualities.

Phytochemical analysis and antifertility potential of Cynodon dactylon in female Wistar rats: A herbal approach towards contraception / 中草药·英文版

Objective: The objective of the present study was to evaluate the antifertility activity of ether (ErCD), chloroform (CeCD) and ethyl alcohol (EyCD) extracts of the whole plant of Cynodon dactylon in female Wistar albino rats. Methods: Acute oral toxicity and an antifertility study were performed in female Wistar rats with two dose levels (200 and 400 mg/kg, orally) of EyCD. The estrogenic and progestogenic effects of EyCD were further observed by administering it to immature Wistar rats by investigations of vaginal cornification, hormonal level, uterus weight, biochemical parameters, histopathology of the uterus and deciduoma formation, respectively. Isolation of EyCD was carried out by Flash Chromatography and isolated fraction was estimated by HPLC. Results: No toxicity with any of the extract was found up to the dose of 2000 mg/kg body weight. EyCD treated rats exhibited maximum reduction in pregnancy (83.33%). Estimation of EyCD on vaginal cornification, estrogen-induced uterotrophic assay and deciduoma model demonstrated vaginal cornification, significant (P < 0.01) increase in uterine weight and uterine proliferation in histopathology and reduced deciduoma formation respectively. Hormonal and biochemical parameters confirmed the above findings indicating estrogenic potential and antiprogestogenic potential of EyCD that might be attributed to the presence of phytoestrogen (apigenin) in EyCD. Conclusion: The results suggested that extracts of C. dactylon possess significant antifertility activity, which is consistent with the literature reported in folk medicine of this plant in fertility regulation.

Anticonceptive and Pharmacognostic Studies on the Seed and Diethyl Ether Extract of RICOM-1013-J

Aims: The anticonceptive effect of the diethyl ether extract of Ricinus communis Linn (RICOM 1013-1) was studied in rats alongside pharmacognostic properties, amino acids content and elemental analysis. Study Design and Methodology: 20 adult female albino Wistar rats were randomized into four experimental groups of five rats each. The first, second and third groups received 3, 10 and 20 mg/kg of diethyl ether extract administered subcutaneously (SC) in divided doses over two days, respectively. The fourth group received 0.1 ml of corn oil for two days and served as control. They were then mated with proven fertile males in a ratio of 3:1 and followed for three gestation periods. The first day of mating was termed day 0 of pregnancy. In addition, amino acid and elemental analyses were undertaken as well as a phytochemical screening of the seed of RICOM-1013-J. Results: At doses of 3, 10 and 20 mg/kg, RICOM-1013-J produced dose-dependent inhibition of pregnancy in female albino Wistar rats over three gestation periods. The total ash value obtained was 3.13 ± 0.26%, while the acid-insoluble and water-soluble ash values were 0.30 ± 0.003% and 0.20±0.003%, respectively. The yield to ethanol was much higher than the water-soluble extract. Essential and non-essential amino acids were not detected. K+, Ca2+ and Mg2+ were present in high concentrations in the following order: defatted undecorticated>undecorticated>decorticated samples. Phytochemical screening indicated the presence of steroids and alkaloids whereas, triterpenes, tannins and flavonoids were absent. Conclusion: The pharmacognostic parameters, including elemental values, provide some preliminary data for identification of this species among many varieties reported in the literature.

Efficacy of CZP3 DNA vaccine immune infertility enhanced by electric pulse method / 中国免疫学杂志

Objective:To analyze the immunocontraceptive efficacy of CZP3 DNA vaccine by electric pulse stimulating and the correlation between antibody level and antifertility. Methods: The mice were immunized with 5, 10, 20 and 50 μg DNA vaccine pcDNA3. 0-CZP3 respectively and were stimulated at leg muscle by electric pulse. Detect the antibody level of each mice group and the highest antibody titer by ELISA. After mated with male mice for three weeks,calculated infertility rate of mice,the average litter size of each immune group and did statistical analysis. Results:Compared with the intramuscular injection alone,the electric pulse stimulation could increase serum antibody titers of mice significantly, the antibody titer of 50 μg group which was stimulated by electric pulse reached 1∶4 000. Moreover,the infertility of the mice increased and the mice average litter size reduced along with the increase of CZP3 antibody level significantly. Statistical analysis showed that the antibody level and the average litter size had a negative correlation. Conclusion:The electric pulse stimulation can improve the immune level and reduce the fertility efficiency of mice significantly,therefore establishes a foundation for further application of CZP3 DNA vaccine in stray dog contraception.

Quantification of anti-fertility compound-Diosgenin concentration in the fenugreek seeds aqueous extract (FSA)

This study aims to build a standardization method for preparation of effective powder from FSA and to quantify diosgenin in FSA. Methodology: One kg of FS were used in this study. Setting: BMS, KOM and KOP, IIUM Kuantan campus. FS were washed with distilled water to exclude any foreign matter, and were then air dried. FS-powder were put in distilled water in a ratio of 1 g of powder in 20 ml of distilled water and were shaken at room temperature for 24 hours. Ten mg of hydrolyzed extract sample was diluted in 10 ml volumetric flask with methanol for 15 minutes. Chromatographic estimation was performed using an equilibrated reverse phase Eclipse XDB-C18 column (particle size 5 µg, 4.6 mm x 150 mm). Results: One gram of FSA extract was hydrolyzed to produce sapogenins and 46.6% was recovered. A calibration curve that was constructed based on five dilutions of diosgenin standard at concentrations of 2, 5, 10, 20, 30 and 50 ppm produced a linear graft (r = 0.999). The concentration of diosgenin in FSA extract as calculated using the regression analysis was found to be 29.66 µg/ml, 13.81 % w/w on dried weight basis. Conclusion: Preparation and standardization of effective powder from FSA are the corner stone of many scientific researches in IIUM and Malaysia. Diosgenin is available in the FSA in adequate concentration. The adequate amount of diosgenin in the FSA will guide us to do further study in the way of preparation of a natural product that can be used in the field of reversible anti-fertility therapy.

Prenatal and developmental toxicity study of meclizine and caffeine combination in female albino wistar rats.

Meclizine and caffeine combination is used for the treatment of morning sickness. Both compounds are teratogenic and caffeine is known to possess anti-fertility activity also. The present study was undertaken to evaluate the reproductive toxic effect of meclizine and caffeine combination. Three doses were taken for the study; low dose (LD; meclizine 3.7 mg/kg and caffeine 3 mg/kg) was selected from commercially available formulation, middle dose (MD; meclizine 37 mg/kg and caffeine 30 mg/kg) and high dose (HD; meclizine 370 mg/kg and caffeine 300 mg/kg). The mixture was administered 1-7 days and 8-14 days for fertility and embryotoxic studies respectively. Laparotomy was done on 10th day of gestation period. Number of implants and corpora lutea were counted, pre and post-implantation losses were determined. In embryo toxicity study fetuses were evaluated for external, skeletal and visceral examination. High dose was removed from both fertility and embryotoxicity studies due to its severe toxicity to the dam. Significant anti-fertility activity was observed at middle dose. Embryotoxicity study showed significant reduction in fetal body weight, body length and body mass index, dam body weight gain on gestation day 14. Absolute kidney weight in MD and absolute and relative spleen weight in both LD and MD were significantly reduced. There was no increase in external or internal congenital anomalies at both LD and MD. The, results suggest that prescription of meclizine and caffeine for morning sickness in early pregnancy should be reviewed carefully.

Antifertility activity of Artemisia vulgaris leaves on female Wistar rats / 中国天然药物

AIM@#To evaluate the antifertility activity of Artemisia vulgaris leaves on female Wistar rats.@*METHOD@#The plant extract was tested for its effect on implant formation at two dose levels, 300 and 600 mg·kg⁻¹, respectively. The effective methanolic plant extract was further studied for estrogenic potency on ovariectomised immature female Wistar rats.@*RESULTS@#The data presented in this study demonstrate the antifertility potential of Artemisia vulgaris methanolic leaf extract, which shows a strong and significant decrease in implant formation (100%), and a strong estrogenic effect resulting in a significant increase in uterine weight in immature ovariectomised rats. These observations suggest that the methanolic extract of Artemisia vulgaris leaves has strong anti-implantation activity and estrogenic activity.@*CONCLUSION@#The methanolic plant extract of A. vulgaris has antifertility activity.

Molecular adjuvant interleukin-33 enhances the antifertility effect of Lagurus lagurus zona pellucida 3 DNA vaccine administered by the mucosal route

It has been shown that cytokines can act as molecular adjuvant to enhance the immune response induced by DNA vaccines, but it is unknown whether interleukin 33 (IL-33) can enhance the immunocontraceptive effect induced by DNA vaccines. In the present study, we explored the effects of murine IL-33 on infertility induced by Lagurus lagurus zona pellucida 3 (Lzp3) contraceptive DNA vaccine administered by the mucosal route. Plasmid pcD-Lzp3 and plasmid pcD-mIL-33 were encapsulated with chitosan to generate the nanoparticle chi-(pcD-Lzp3+pcD-mIL-33) as the DNA vaccine. Sixty female ICR mice, divided into 5 groups (n=12/group), were intranasally immunized on days 0, 14, 28, and 42. After intranasal immunization, the anti-LZP3-specific IgG in serum and IgA in vaginal secretions and feces were determined by ELISA. The results showed that chi-(pcD-Lzp3+pcD-mIL-33) co-immunization induced the highest levels of serum IgG, secreted mucosal IgA, and T cell proliferation. Importantly, mice co-immunized with chi-(pcD-Lzp3+pcD-mIL-33) had the lowest birth rate and mean litter size, which correlated with high levels of antibodies. Ovaries from infertile female mice co-immunized with chi-(pcD-Lzp3+pcD-mIL-33) showed abnormal development of ovarian follicles, indicated by atretic follicles and loss of oocytes. Our results demonstrated that intranasal delivery of the molecular adjuvant mIL-33 with chi-pcD-Lzp3 significantly increased infertility by enhancing both systemic and mucosal immune responses. Therefore, chi-(pcD-Lzp3+pcD-mIL-33) co-immunization could be a strategy for controlling the population of wild animal pests.

Polymeric nanoparticle formulation of Octapeptide (NP-OP): In vitro release and in vivo effect in common marmosets, Callithrix jacchus Linn.

yielded an average particle size of 120 nm with 70% encapsulation-efficiency. In vitro release profile of NP-OP showed sustained release of OP for 21 days. In vivo anti-fertility studies were conducted in marmosets. Results indicated that control animals conceived in the same cycle while two of three treated animals failed to conceive in treatment cycle. The in vivo studies thus corroborate with in vitro release of OP, demonstrating its anti-fertility activity in 66% of animals.

Seeds of mimusops elengi linn- an antifertility drug.

The study involves a standard pharmacological model to screen antifertility activity of seeds of Mimusops elengi Linn. When the aqueous powdered drug (2gm/body weight) was administered to male albino rats has proved to be an effective male contraceptive drug. The activity was confirmed by significant decrease in sperm count, biochemical assays so also through histopathological investigations. Hence seeds of Mimusops elengi can be a reliable herbal option after the necessary clinical trials.

An electron microscopic study of the antifertility potential of rosemary (Rosmarinus officinalis L) in male albino rats / Estudio de microscopía electrónica del potencial antifertilidad del Romero (Rosmarinus officinalis L) en ratas macho albinas

The present work was aimed at studying the antifertility potential of the commonly used herb, rosemary in the male albino rats using electron microscopy as the method of investigation. Ethanolic extract of the rosmary prepared and administered orally in two different doses for a period of three months to the animals. At the end of the experiment animals were sacrificed and testes removed. Sections for the electrone microscopy prepared and changes were observed. The present results showed evident microscopic changes in the testis of the animals received higher dose of the drug. Most of the seminiferous tubules were compressed, having irregular basement membrane and devoid of any spermatogenic cells. The present work revealed a clear morphological evidence of the dose dependent antifertility potential of the rosemary in the male albino rats.

El trabajo tuvo como objetivo estudiar el potencial anti-fertilidad de la hierba de uso común, el romero, en ratas albinas macho utilizando microscopía electrónica como método de investigación. El extracto etanólico del romero se preparó y administró por vía oral a los animales en dos dosis diferentes durante un período de tres meses. Los animales experimentales se sacrificaron y se retiraron sus testículos. Se prepararon secciones para microscopía electrónica y se observaron los cambios. Los resultados mostraron cambios microscópicos evidentes en los testículos de los animales que recibieron una dosis mayor del medicamento. La mayoría de los túbulos seminíferos se observaron comprimidos, con una membrana basal irregular y carente de células espermatogénicas. El presente trabajo revela una clara evidencia morfológica de una posible anti-fertilidad dependiente de la dosis del romero administrada en las ratas albinas macho.

Pharmacological evaluation of anti-fertility activity of ethanolic extract of Jatropha gossypifolia leaf in female albino mice

Objective: Anti-fertility activity of ethanolic extract of Jatropha gossypifolia leaf in female albino mice. Methods: Jatropha gossypifolia leaf extract, when administered orally, altered the estrous cycle pattern in female mice, prolong the length of estrous cycle with significant increase in the duration of diestrus stage and reduced significantly the number of litters in albino mice. Treatment of mice with extract of 250 and 450 mg/kg body weight/day for 21 days caused a prolonged estrous cycle with significant increase in the duration of diestrus phase and elongation of estrus stage in treatment with higher dose (450 mg/kg body weight/day). Results: The analysis of the principal hormones involved in estrous cycle regulation showed that the plant extracts altered gonadrotrophin release (LH, FSH and prolactinn) and estradiol secretion. Conclusions:The results indicated the anti-fertility effect of Jatropha gossypifolia leaf extract in female albino mice.

Antifertility effect of madhuca latifolia (ROXB.) macbride seed extract.

The study involves a standard pharmacological model to test antifertility activity of seeds of Madhuca latifolia (Roxb.) Macbride. When the aqueous powdered drug (2gm/body weight) was administered to male albino rats has proved to be an effective antifertility drug. The activity was confirmed by significant decrease in sperm count, biochemical assays so also through histopathological investigations. Hence seeds of Madhuca latifolia can be a reliable herbal option after the necessary clinical trials.

Isolation and evaluation of antifertility activity of total alkaloids from leaves of aegle marmelos in male albino rats (rattus norvegicus).

Aegle marmelos (Rutaceae) is being used in traditional medicine treatments, such as for intermittent fever, intestinal ailments, fertility control etc. It has been proved to be effective against several major diseases including cancer, diabetis and cardiovascular diseases. Although the plant is a well known male antifertility plant, till today only the crude extracts of the plant were screened for antifertility activity in male rats. The plant is rich in alkaloid content and Aegelenine, Marmeline and Skimmianine, are some of the alkaloids isolated so far, showed variety of pharmacological activities. In view of these facts, in the present study, total alkaloids have been isolated from leaves of A. marmelos and their effect on fertility of adult male albino rats (Rattus norvegicus) was investigated. Three different doses 20, 40, 80 mg/kg body weight of total alkaloids were orally administered to mature male albino rats (Wistar strain) of proven fertility (235-2450gr) for 60 days. On day 61, all the animals were sacrificed and the fertility and safety parameters were studied. Weights of all the major reproductive organs, accessory glands and sperm counts were significantly decreased in dose dependent manner suggesting the antifertility activity and serological parameters showed no significant changes in treated animals at the tested dose levels indicating the safety of long-term use of total alkaloidal fraction of A. marmelos.

Antifertility effects of herbs: Need for responsible reporting.

Evaluation of herbs for antifertility effects has been in progress worldwide for several decades to identify effective and safe substances for control of population explosion. Inspite of availability of internationally accepted guidelines for the assessment of reproductive toxicity/antifertility potential of test substances, many published articles, on critical review, seem to lack reproducibility and are thus likely to mislead both the scientific community and the general public. This paper, while emphasizing the importance of generating authentic toxicity/safety information on acclaimed medicinal herbs, spells out existing pitfalls in such studies, and explores some control measures worth considering in times to come.

Effect of ethanol extract of whole plant of Trichosanthes cucumerina var. cucumerina L. on gonadotropins, ovarian follicular kinetics and estrous cycle for screening of antifertility activity in Albino Rats / Efectos del etanol extraído de la planta completa de Trichosanthes cucumerina var. cucumerina L. sobre gonadotropinas, cinética folicular ovárica y ciclo para screening de actividad antifertilidad en Ratas Albinas

Ethanol extract of whole plant of Trichosanthes cucumerina L. var. cucumerina was evaluated for antiovulatory activity in adult rats. The ethanol extract at the doses 200 and 400mg/kg body weight (orally) affected the normal estrous cycle showing a significant increase in estrus and metestrus phases and decrease in diestras and proestrus phases. The extract also significantly reduced the number of healthy follicles (Class I-Class VI) and corpora lutea and increased the number of regressing follicles (Stage IA, Stage IB, Stage IIA, and Stage IIB). The protein and glycogen content in the ovaries were significantly reduced in treated rats. The cholesterol level was significantly increased, whereas, the enzyme activities like 3b-HSD and 17b-HSD were significantly inhibited in the ovary of treated rats. Serum FSH and LH levels were significantly reduced in the treated groups were measured by RIA. In acute toxicity test, neither mortality nor change in the behavior or any other physiological activities in mice were observed in the treated groups. In chronic toxicity studies, no mortality was recorded and there were no significant differences in the body and organ weights were observed between controls and treated rats. Hematological analysis showed no significant differences in any of the parameters examined (RBC, WBC count and Hemoglobin estimation). These observations showed the antiovulatory activity of ethanol extract of whole plant of Trichosanthes cucumerina L. var. cucumerina in female albino rats.

El extracto de etanol de toda la planta de Trichosanthes cucumerina var. cucumerina (L.) se evaluó en cuanto a su actividad antiovulatoria en ratas adultas. El extracto de etanol en dosis de 200 y 400mg/kg de peso corporal (oral) afectó el ciclo normal estral, mostrando un aumentó significativo en las fases de estro y metaestro y la disminución de las fases de diestro y proestro. El extracto también redujo significativamente el número de folículos sanos (Clase I=Clase VI) y cuerpo lúteo y aumentó el número de folículos en regresión (etapa I, etapa IB, etapa II y etapa IIB). La proteína y el contenido de glucógeno en los ovarios se redujeron significativamente en las ratas tratadas. El nivel de colesterol aumentó significativamente, mientras que, actividad de las enzimas 3b-HSD y 17b-HSD se inhibió significativamente en el ovario de ratas tratadas. FSH sérico y los niveles de LH se redujeron significativamente en los grupos tratados y medidos por RÍA. En la prueba de toxicidad aguda, no hubo mortalidad ni cambio en el comportamiento fisiológico o de cualquier otra actividad en los grupos tratados de ratas. En estudios de toxicidad crónica, no se registró mortalidad y no hubo diferencias significativas en el peso corporal o el peso de los órganos entre los controles y las ratas tratadas. Los análisis hematológicos no mostraron diferencias significativas en ninguno de los parámetros examinados (eritrocitos, recuento de glóbulos blancos y estimación de hemoglobina). Estas observaciones mostraron la actividad antiovulatoria del extracto de etanol de toda la planta de Trichosanthes cucumerina var. cucumerina en ratas albinas hembras.

SODIUM MORRHUATE:APPLICATION AND EXPLORATION / 中国海洋药物

This paper reviewed the history of Sodium Morrhaute,summarized the action mechanism and clinical use of Sodium Morrhuate as sclerosant and hemostyptic, and described the research and application of Sodium Morrhuate as the drug for birth control. The further applications for antitumorigenesis and antifertility in the medical field were also discussed.

Antifertility effect of “terpinnen-4-OL” in hamsters.

“Terpinen-4-ol” a cyclic terpene alcohol isolated from oil of Zingiber cassumunar Roxb., was tested for its antifertility effect in hamsters. The fertility test was performed according to the MB 55 WHO protocol (52 days of treatment). The results of the experiment revealed that at dose levels of 100 and 200 mg/kg, the number of epididymal sperm decreased significantly, by 65 and 74 percent respectively. More than 90 percent of the sperm abnormal morphology, especially at the midpiece portion of the tail; motility was also markedly depressed. The sperm motility rate of the group receiving 100 mg/kg varied from 0 to 40 percent compared with 74 percent of the control group. The sperm motility rate was totally zero in the group treated with 200 mg/kg. When the males were mated with females, the pregnancy rate of the mated females was 0 percent (with both low and high doses) compared with 100 percent for the control group; there was no evidence of abnormal, live or dead fetuses or other parameter connected with pregnancy. Blood levels of FRH, LH and testerone were not significantly changed. Histological study of the testis revealed no interference with spermatogenesis at all doses tested. Moreover, no significant alteration in body weight, or in adrenal gland, testicular and accessory sex organ weights except for the weights of the cauda and total epididymis which were significantly decreased. When the modified MB 55 protocol (22 days of treatment) was applied, exactly similar results, as described above were obtained. When Terpinen-4-ol was tested for its antifertility effect in female hamsters, no significant alteration on fertility parameters, namely pregnancy rate, number of live fetuses including abnormal fetuses, implantation sites and corpus luteum of pregnancy was observed. These results indicate with certainty that this compound possesses potent antipermatogenesis and antifertility effects in males only. Further study to again more information would certainly be worthwhile and may lead to the development of a male contraceptive for human use in the future.

MECHANISM OF ANTIFERTILITY BY GOSSYPOL OPTICAL ISOMERS IN RATS / 解剖学报

12 kinds of histochemical methods were adopted to evaluate the effects of gossypol optical isomers on the reproductive system of Wistar rats, following the administration of (?) and (+)-gossypol at a dosage of 30mg/kg daily, and(-)-gossypol at a dosage of 15 mg/kg daily, respectively for 2 or 3 weeks. (+)-gossypol has no an tifertility activity. (?) and (-)-gossypol caused kinetic disorder of spermatogemsis, and inhibited turnover of nuclear basic protein of the late, elongated spermatids in the testis. No changes in G6PDH and 30-HSDH histochmical reaction in Leydig cells, and ALP in the basement membrane of epididymis were observable reflecting that gossypol has no effect on synthesis and secretion of testosterone. Spermatozoa in the epididymis of rats treated with (?) and (-)-gossypol exhibited head and tail separation. The formazan deposits of reaction of LDH, LDH-X, MDH and ICDH located in sperm mitochondria appeared to become coarse and to reduce in quantity after (?) and (-)-gossypol treatment.