ReProComet: a new in vitro method to assess DNA damage in mammalian sperm.

The increasing request of chemical safety assessment demands for the validation of alternative methods to reduce the resort to animal experimentation. Methods that evaluate reproductive toxicity are among those requiring the largest use of animals. Presently, no validated in vitro alternative exists for the assessment of reproductive toxicity. Mammalian sperm are sensitive targets of DNA-reactive chemicals, which form premutagenic adducts. Here, we propose a new method based on comet assay to detect DNA damage induced by potential germ cell mutagens in bull sperm available from assisted reproduction practices. In somatic cells, chemical-induced adducts can be revealed by comet assay that detects DNA breaks produced during adduct repair. Mature sperm, however, are devoid of repair enzymes, and adducts are processed only after fertilization. For this reason, comet assay is not sensitive to detect DNA lesions induced in sperm by most chemicals. To overcome such limitation, we developed a modified comet assay based on the addition of a protein extract from HeLa cells to agarose-embedded sperm on microscopic slides. To test the method, sperm were treated in vitro with methyl methanesulfonate (MMS) or melphalan (MLP) and comet assay was conducted both with and without protein supplementation. No effect of MMS or MLP was detected without protein supplementation; on the contrary, a clear-cut dose-dependent effect was measured after addition of the cell extract. These results represent a proof of concept of a novel in vitro mutagenicity test on sperm that could offer a promising approach to complement previously validated in vivo germ cell genotoxicity assays.     

不同繁殖材料对半夏产量的影响

实验表明:半夏有性繁殖种子田间出苗率为0～25%,且实生苗当年不能形成商品;无性繁殖则以珠芽和小块茎作种明显优于大块茎。     

中西医结合要为妇女生殖健康创新页

45年来,复旦大学妇产科医院在中西医结合临床及科研方面取得了可喜的成就.针刺促排卵的研究,通过观察血雌激素和中枢β-内啡肽(β-EP)的关系,提出了针刺通过耗竭中枢β-EP而促排卵的新认识,认为针刺促排卵的主要适应症是青春期月经失调,并将针刺成功地应用于治疗神经性厌食;补肾化痰的俞氏温补方治疗多囊卵巢综合征(polycystic ovary syndrome, PCOS)被证实是通过提高血促卵泡激素(follicle stimulating hormone, FSH)并诱发排卵而起作用;益肾化瘀的天癸方是通过降低患者及动物模型的雄激素和胰岛素,引起以中枢阿片促黑激素皮质素原、神经肽Y和瘦素受体为主的神经-内分泌-代谢网络调节而发挥促排卵和减肥作用,对高胰岛素型PCOS有良效,并在研究中提出了对PCOS分类的新设想;益肾清火的更年春通过提高中枢与周围组织的雌激素受体及其mRNA表达来调节围绝经期妇女的神经-内分泌-免疫-代谢网络而获优越疗效.这些研究表明,疾病是外环境影响下患者生命网络的某些主脉失调所致,而且亚健康状态的妇女具有初步网络失控的现象,中西医结合将为女性生殖医学研究找到捷径,调控生命网络是防病治病的主要思路.     

Analysis of Implantation in Assisted Reproduction Through the Use of Serial Human Chorionic Gonadotropin Measurements

Purpose: Our purpose was to examine implantation of singleton pregnancies achieved following various assisted reproductive technologies (ARTs) through the appearance and rising titers of serum human chorionic gonadotropin (hCG) levels. Methods: A total of 114 singleton pregnancies resulting from in vitro fertilization and intrauterine insemination was analyzed. Patients were divided into five groups according to the type of ovarian stimulation protocol [gonadotropin stimulation with/without the use of gonadotropin-releasing hormone agonist (GnRHa), long protocol, or flare-up technique] and to the day of embryo transfer (day 2 or day 3 after oocyte retrieval). Serial serum hCG levels were measured between 10 and 25 days after fertilization and log-transformed. Linear regression analyses were performed and extrapolated to hCG = 10 mIU/ml (hCG ₁₀ ), which was used as an estimate of detectable implantation. The slopes of the regression lines were used to estimate the rising speed of hCG. Results: There were no significant differences in the days of hCG in maternal serum to reach 10 mIU/ml (implantation) or in the slopes of the regression lines for all five studied groups. Conclusions: The appearance of hCG in maternal serum was used to assess the time of clinically detectable implantation. Furthermore, because hCG production is a marker of trophoblastic activity, its serum doubling time was used as an indicator of embryo quality. Results showed that in various ART protocols with and without GnRHa, there were no significant differences in implantation time or embryo quality. Embryo development in early pregnancy follows a preprogrammed-timing schedule and depends mainly on the embryonic age of the health, successfully implanted conceptus.     

Recurrent pregnancy loss due to familial and non-familial habitual molar pregnancy.

OBJECTIVES: To present a series of women with recurrent molar pregnancies, including rare familial cases, and discuss etiology and treatment options. METHODS: We performed a detailed clinical evaluation and pedigree analysis of five Egyptian women with recurrent pregnancy loss due to molar pregnancy. RESULTS: The women had a history of four to nine consecutive hydatidiform moles but of no viable pregnancies. Two of the women had molar pregnancies with different husbands who themselves had viable offspring from previous wives; and three of them, who belonged to a family with extensive intermarriage, had a pedigree consistent with an autosomal recessive maternal-effect mutation. CONCLUSIONS: Recurrent pregnancy loss due to habitual molar pregnancy is uncommon and familial cases are extremely rare. The etiology of this disorder is not well understood but likely results from a maternal-effect mutation. Management options are limited, especially for couples who desire to have their own genetic offspring.     

Neuroactive Properties of Reproductive Steroids

Migraine is 3 times more common in postpubertal women than in men. Migraine is frequently exacerbated perimenstrually and commonly occurs exclusively at that time. It is often benefited by pregnancy and menopause. Estrogen withdrawal has been implicated as a mechanism for triggering migraines. The mechanism, however, is not well understood. Reproductive steroids have neuroactive properties that can modulate neuronal morphology and physiology. Increasing evidence suggests that circulating reproductive steroid levels regulate the balance of neuroexcitatory and neuroinhibitory activities in some brain regions by influencing synaptic plasticity. Estrogen has neuroexcitatory, whereas progesterone has neuroinhibitory, effects in most preclinical and clinical models. Several neurotransmitter systems that are implicated in migraine vary with reproductive steroid levels during the reproductive cycle. Estrogen stabilization may provide effective treatment in susceptible women, especially for catamenially exacerbated migraine.     

Endogenous opioid-immunoreactive neurons of the ventromedial hypothalamic nucleus concentrate estrogen in male and female rats

Estrogen stimulates expression of proenkephalin mRNA in neurons of the hypothalamic ventromedial nucleus, and evidence is accumulating that synaptic release of one of the peptide end products, met-enkephalin, influences events that regulate reproductive behavior. To address the question of whether estrogen acts directly on neurons that synthesize met-enkephalin or indirectly through a separate neuronal population, we combined estrogen autoradiography with endogenous opioid peptide (EOP) immunohistochemistry. In agreement with previous studies, the ventrolateral subdivision of the hypothalamic ventromedial nucleus was densely packed with EOP-immunoreactive cells. In males, 48% of the estrogen-concentrating cells of the ventrolateral subdivision of the hypothalamic ventromedial nucleus contained EOP, and, in females, 27% of the estrogen-concentrating cells contained EOP. These findings indicate that estrogen acts directly on neurons that express EOP and suggest a mechanism that underlies sexually differentiated reproductive behavior.     

Cellular localization of immunoreactive epidermal growth factor during Wolffian duct differentiation of the fetal mouse

Previous studies from this laboratory indicated a role for epidermal growth factor (EGF) in androgen-dependent male reproductive tract differentiation of the fetal mouse. Expression of an EGF-like protein during Wolffian duct differentiation was indicated from the determinations by radioimmunoassay (RIA) and radioreceptor assay. To further characterize the protein and to assess its role in male sexual differentiation, expression of the protein has been analyzed by Western blot assay and its tissue-specific cellular expression has been determined by immunocytochemical assay in the present study. Western blot analysis of the 18-day fetal male reproductive tract detected an immunoreactive band of the predicted 6-kDa size. Immunocytochemical analysis also detected EGF-specific immunostaining in the Wolffian duct derivatives. At day 18 of gestation, the staining was localized predominantly in the epithelial nuclei of the Wolffian duct derivatives whereas at days 14 and 16 of gestation, the staining was equally distributed in the mesenchymal and epithelial sites of the Wolffian duct derivative. The intensity of the staining increased with progression of differentiation during the 14th–18th days of gestation. Prenatal exposure to the antiandrogen flutamide significantly reduced the immunostaining of the duct. Thus, a role for EGF in Wolffian duct differentiation is indicated.     

Reproductive health knowledge and implications: A study in Nigeria

Forty-five percent of Nigeria's population is under age 15. This study employed a questionnaire to learn the level of reproductive health knowledge of 416 male and female Nigerian students ages 10 through 16.

The Chi-square statistical procedure was used as demographic characteristics were related to reproductive health knowledge among the pre-teen and teenage youth. Statistically significant (p #lt 0.05) relationships were found between variables such as knowledge of reproduction, family planning, and AIDS when each was compared separately to age, gender, living situation while at school, school club memberships, and so on. With respect to reproduction and related topics, older teenagers indicated more knowledge than pre-teenagers, girls more than boys, and those living in dormitories more than those living at home or in other housing while attending school.

Television ranked first in terms of students' sources of information on reproduction, pregnancy, birth control, and AIDS. Newspapers ranked second, radio third, teachers fourth, and parents fifth. Perhaps the most striking data are the “I don't know” responses with respect to AIDS. Thirteen percent had not heard of AIDS, 27 percent did not know how AIDS is transmitted, 29 percent did not know the seriousness of AIDS, 37 percent did not know how to avoid AIDS, and 14 percent did not know that a mother with AIDS might infect her baby. Recommendations are made for increasing health services and education concerning reproductive health for pre-teen and teenage students.