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Effect of different culture techniques used to induce capacitation on the chromatin stability of human sperm.

To validate an in-vitro bioassay for assessing chromatin stability of human sperm, 38 semen samples from infertile men were studied using sodium dodecyl sulphate, an anionic detergent which disorganizes only the cytoplasmic membrane. Assay sensitivity was 50 sperm, whilst the within- and between-assay variation, and the between-observer variation were found to be within the accepted range for this type of bioassay. The influence of different in-vitro treatments currently used in some clinical assisted fertilization programmes was evaluated: a destabilizing effect occurred in Grade I (stable) and Grade III (swollen) sperm. In the former, all treatments reduced stable sperm; in the latter, a significant (P less than 0.001) increase in swollen sperm was shown with two methods that used Ham's F-10 as culture medium. Different chromatin patterns found in the treated sperm suggest the possibility that the recovered samples could be modified compared to their status at the time of isolation.

Biological Assay↗

The role of medical problems and behavioral risks in explaining patterns of prenatal care use among high-risk women.

OBJECTIVE: To examine the associations between maternal medical conditions and behavioral risks and the patterns of prenatal care use among high-risk women. DATA SOURCE/STUDY DESIGN: Data on over 25,000 high-risk deliveries to African American and white women using multinomial logistic regression to predict the odds of adequate-plus care relative to three other categories of care. DATA COLLECTION/EXTRACTION METHODS: Data were extracted from records maintained by the University of Florida/Shands Hospital maternity clinic on all deliveries between 1987 and 1994; records for white and for African American women were subset to examine racial differences in medical conditions, health behaviors, and patterns of prenatal care use. PRINCIPAL FINDINGS: Net of sociodemographic and fertility-related characteristics, African American and white women with late antepartum conditions and hypertension problems had significantly higher odds of receiving adequate-plus care, as well as no care or inadequate care, relative to adequate care. White women with gynecological disease and medical/surgical problems were significantly less likely to receive no care or inadequate care, as were African American women with gynecological disease. CONCLUSIONS: Maternal medical conditions explain much but not all of the adequate-plus prenatal care use. More than 13 percent of African American women and 20 percent of white women with no reported medical problems or behavioral risks used adequate-plus care. Additional research is needed to understand this excess use and its possibilities in mediating birth outcomes.

Adolescent↗

Localization of molecules related to cholinergic signaling in eggs and zygotes of the sea urchin, Paracentrotus lividus.

In recent years, the finding of a complete set of molecules related to the cholinergic neurotransmitter system in male gametes of different invertebrate and vertebrate species has opened the question of the possible involvement of this signaling system in the regulation of intracellular ion change, functional to sperm motility and probably also to interaction with the egg. In this work, we localized molecules immunologically related to muscarinic acetylcholine receptors (mAChRs) in sea urchin (Paracentrotus lividus) eggs and zygotes, by use of the monoclonal antibody M35. The ultrastructural analysis of immunoreactivity revealed that the localization of M35-positive molecules is different between unfertilized and fertilized egg. In the unfertilized eggs, immunoreactive molecules were localized inside and associated to the membrane of vesicles, scattered in a thick zone of the cortical cytoplasm. Following fertilization, the oolemma was decored along the surface, and positive vesicles were seldom seen in the cytoplasm. At the surface of fertilized eggs, evident pinocytotic vesicles, recognized by the surrounding coat, indicate that the positive sites correspond to receptorial activity, able to rescue ligand portions. The presence of a cholinergic signaling pattern was also suggested by the presence of molecules immunologically related to acetylcholinesterase (AChE), performing AChE enzyme activity, detected by both immunofluorescent and histochemical methods in membranous organelles belonging to the cortical region of unfertilized eggs.

Acetylcholinesterase↗

Contraceptive use, pregnancy and fertility patterns among single American women in their 20s.

Eighty-two percent of never-married American women aged 20-29 have had sexual intercourse; black women are somewhat more likely than white women to have had intercourse. In all, 53 percent of never-married women in this age-group had intercourse at least once in the four weeks preceding the 1983 National Survey of Unmarried Women. Black women are more likely than white women to have done so (62 percent compared with 51 percent). Nearly all of the women who ever had intercourse have used a contraceptive method at some time; 78 percent practiced contraception at the time of their most recent intercourse. A high proportion did not start using birth control until some time after first intercourse, however: On average, the delay between first coitus and first contraceptive use was eight months, and one-fifth of the respondents said that they began using a method only after their first pregnancy. Most of the women who did use a method at the time of first intercourse relied on the condom or withdrawal; in contrast, about two-thirds of white women and three-quarters of black women now rely on the pill, IUD or sterilization. Eighty-six percent of the women who had intercourse in the four weeks before the interview were current users--88 percent of the white women and 77 percent of the black women. Catholic women are no less likely than others to have ever had intercourse, to be currently sexually active or to be using contraceptives. However, Catholic women who receive communion at least once a week are less likely to be sexually active and substantially less likely to use medical contraceptive methods. Women who consider themselves very religious are less likely to be sexually active, but the sexually active among them are about as likely as others to use contraceptives. Better-educated women are much more likely than less-educated women to practice contraception, and women who work outside of the home are more likely than those who do not to use contraceptives. Thirty-three percent of unmarried 20-29-year-olds have had at least one pregnancy (about 40 percent of those who have ever had intercourse). Thirty-two percent of sexually active white women have been pregnant, compared with 70 percent of comparable black women. Furthermore, whereas 14 percent of white 20-29-year-olds have had an out-of-wedlock birth, 62 percent of black women have done so.(ABSTRACT TRUNCATED AT 400 WORDS)

Abortion, Induced↗

Normal female phenotype and ovarian development despite the ovarian expression of the sex-determining region of Y chromosome (SRY) in a 46,XX/69,XXY diploid/triploid mosaic child conceived after in vitro fertilization-intracytoplasmic sperm injection.

CONTEXT: Diploid/triploid mosaicism (mixoploidy) is a rare chromosomal abnormality characterized by mental and growth retardation, hypotonia, and dysmorphic features such as facial asymmetry, low-set ears, and syndactyly. All 46,XX/69,XXY cases fall into three phenotypic groups: male with testicular development, ovotestis disorder of sex development (DSD), or undervirilized male DSD. All phenotypic females with diploid/triploid mosaic reported so far had 46,XX/69,XXX karyotype. PATIENT: We report an 8-year-old girl conceived after in vitro fertilization-intracytoplasmic sperm injection with normal internal/external genital and ovarian development despite 46,XX/69,XXY mosaicism and normal expression of sex-determining region of Y chromosome (SRY) in her gonads. INTERVENTION: Because of the increased risk of gonadoblastoma resulting from Y chromosome mosaicism, her ovaries were removed by laparoscopy. Ovarian tissue was analyzed histologically as well as by fluorescence in situ hybridization, PCR, and RT-PCR amplification to determine the localization of Y chromosome and expression of SRY and DAX1 mRNA. Methylation-specific PCR was used to assess the inactivation pattern of X chromosomes. RESULTS: By laparoscopy, internal female genital anatomy appeared to be normal. Cytogenetic and molecular methods confirmed the presence of intact and functionally active Y chromosome in the ovary. Strikingly, histological assessment of the gonads showed normal ovarian architecture with abundant primordial follicles despite the presence of the Y chromosome in ovarian follicles and the expression of SRY mRNA in gonadal tissue. CONCLUSION: This case illustrates that normal ovarian development is possible in the presence of Y chromosome in ovarian follicles and despite the expression of SRY in ovarian tissue. Furthermore, this is the first documented case of mixoploidy after in vitro fertilization-intracytoplasmic sperm injection and the only phenotypic female with 46,XX/69,XXY karyotype.

Child↗

The flagellar apparatus of spermatozoa in fish. Ultrastructure and evolution.

Chondrichthyes possess an evolved type of spermatozoa. Their flagellar apparatus is characterized by the presence of flagellar roots which form the axis of the midpiece, and the existence of one or two lateral elements associated with the axoneme. Osteichthyes, mainly teleosteans, show a great diversity of spermatic forms. The primitive spermatozoon with a 9 + 2 pattern flagellum is common. The primitive spermatozoon has evolved along different lines. The spermatic diversity which results from this is mainly evident in the structure of the flagellar apparatus. In the animal kingdom the primitive spermatozoon with a 9 + 2 pattern flagellum, present in primitive metazoa, is retained in phyla where external fertilization is maintained. The main evolutionary tendencies--elongation, aflagellarity or biflagellarity--are generally connected with the acquisition of internal fertilization. These evolutionary tendencies are found in teleosteans. It is not possible to link aflagellarity or biflagellarity of the gamete in certain fishes to this method of fertilization. Only the elongation of the spermatozoon is connected, in certain cases, with internal fertilization, but this cannot be taken as general.

Animals↗

Structured antedependence models for genetic analysis of repeated measures on multiple quantitative traits.

Simultaneous analysis of correlated traits that change with time is an important issue in genetic analyses. Several methodologies have already been proposed for the genetic analysis of longitudinal data on single traits, in particular random regression and character process models. Although the latter proved, in most cases, to compare favourably to alternative approaches for analysis of single function-valued traits, they do not allow a straightforward extension to the multivariate case. In this paper, another methodology (structured antedependence models) is proposed, and methods are derived for the genetic analysis of two or more correlated function-valued traits. Multivariate analyses are presented of fertility and mortality in Drosophila and of milk, fat and protein yields in dairy cattle. These models offer a substantial flexibility for the correlation structure, even in the case of complex non-stationary patterns, and perform better than multivariate random regression models, with fewer parameters.

Animals↗

Fertility transition in Bangladesh: understanding the role of the proximate determinants.

Bangladesh has been passing through a crucial phase of fertility transition. The level of fertility declined dramatically during the early 1990s without any remarkable improvement in socioeconomic and health status, and then remained constant at a high level of 3.3, despite the increased use of contraception. Such fertility transition can be traced to variations in one or more of the proximate or direct determinants of fertility. This paper critically analyses the fertility levels in Bangladesh with a view to exploring the possible explanations of fertility decline in the 1990s and then its stabilization. The main focus of the study is to examine the role of the major proximate determinants of fertility in bringing about the change in fertility level in Bangladesh. The data for the study come from a series of nationally representative surveys over the period of 1975 to 1999-2000. The analysis indicates that fertility has temporarily ceased to decline in recent years due to the 'tempo' effect of high past fertility, but in general a declining trend in fertility is underway. The analysis suggests that the fall in fertility is consistent with the underlying trends in most important proximate determinants of fertility. In recent years contraception has emerged as the highest fertility reducing factor in Bangladesh and its effect is greatest in middle and older age groups. Although until the early 1990s postpartum lactational infecundability was the most important and strongest fertility reducing factor in Bangladesh, in recent years its fertility inhibiting effect has gradually decreasing owing to the declining trend in the lactational infecundability period. The analysis reveals that although the fertility reducing effect of the marriage pattern is increasing, its effect is offset by the declining trend in the lactational infecundability period. A review of these two variables suggests that their effect cannot be raised much for prevailing socioeconomic and cultural reasons, and any future reduction in fertility in Bangladesh may be largely dependent on increased use of effective birth control methods.

Adolescent↗

Stage I seminoma of the testis: a bi-institutional retrospective analysis of patients treated with radiation therapy only.

OBJECTIVE: To analyse relapse patterns, toxicity and second malignancy in patients with stage I pure germ cell testicular tumours, treated in 1970-1999. PATIENTS AND METHODS: In all, 487 patients received irradiation after surgery to the infra- (407, 83.5%) or infra- and supra-diaphragmatic volumes (80, 16.5%). Treatment-related toxicity was classified according to previous criteria and fertility investigated in 246 men. Second malignancies were identified by retrospective analysis of clinical records or telephone interviews in men who no longer needed a long-term follow-up. RESULTS: The 10-year overall survival was 97% (98% and 96%, respectively, for the aortic nodes only, or aortic and iliac nodes, i.e. the 'dog leg' field) and disease-free survival was 94%. Twenty-one patients relapsed (five with a true 'in-field' recurrence, nine progressed to the mediastinum, and seven had disseminated disease). Acute toxicity was mainly gastrointestinal, with 7.6% classified as grade II. In all, 73 men achieved paternity after irradiation; nine did not but had normal sperm. Second malignancies were diagnosed in 16 (3.3%) men. CONCLUSION: Para-aortic irradiation may be used safely in patients with stage I seminoma and undisturbed testicular drainage, with equivalent results to the 'dog-leg' group; these unrandomized data confirm the lower toxicity and equivalent survival rates of this treatment.

Adolescent↗

Localization of cystathionine beta synthase in mice ovaries and its expression profile during follicular development.

BACKGROUND: In vitro fertilization (IVF) researches have suggested that cystathionine beta synthase (CBS) is involved in oocyte development. However, little is known about the regional and cellular expression patterns of CBS in the ovary. The purpose of this study was to analyze the localization of CBS in mice ovaries and to investigate the expression profile during follicular development. METHODS: We used in situ hybridization and immunohistochemical analysis to determine CBS expression in the ovaries of female Balb/c mice. Then the follicles were collected from F1 (C57BL x Balb/c) mice and cultured in vitro. With the method of semi-quantitative RT-PCR, we also investigated the expression profile of CBS during follicular development. RESULTS: CBS was absent in the oocytes, although it was ubiquitously expressed in the ovary with the strongest expression in follicular cells at all stages. In late antral follicles, CBS expression was markedly higher in granulosa cells located close to the antrum and in cumulus cells around the oocyte. The semi-quantitative RT-PCR showed that CBS mRNA was detected in follicles at all stages in vitro. In cumulus-oocyte complexes superovulated, CBS expression also increased rapidly. CONCLUSIONS: CBS was located mainly in the follicular cells in the ovaries. The level of CBS expression is high in follicles during folliculogenesis in mice. Differences in the CBS expression profile between oocyte and follicular cells suggest a role for CBS as a mediator in interactions between oocyte and granulosa cells.

Animals↗

[Long acting hormonal contraception].

Long-acting hormonal contraception has been widely accepted in different cultures, where the use of injectables and implants is preferred over other methods of fertility control. The administration of synthetic progestogens in combination with estrogens as injectables suppresses ovulation, allowing the control of the endometrial bleeding too. The use of injectables containing only progesterone constitutes a 2 to 6 months contraceptive method with high effectiveness and easy administration, although its major disadvantage is the alteration of the endometrial bleeding pattern, which is the most common cause of discontinuation. The group of long-acting injectables includes the development of microencapsulated steroids, whose structure gives them the property of maintaining controlled liberation of the drug. Recently subdermal contraceptive implants (Norplant) have appeared, based on the subcutaneous sustained liberation of levonorgestrel. The daily drug dose is enough to inhibit ovulation in the initial months of use and modifies the cervical mucus in the subsequent months. Norplant has been shown to be efficient in preventing pregnancy, but main secondary effect is bleeding pattern disturbance during the menstrual cycle. The Capronor constitutes a new system of contraceptive implant which is different from Norplant, where the capsules containing levonorgestrel are of unbiodegradable material, and Capronor is formed by a biodegradable polimerus. Finally, among the contraceptive methods that have been developed to date there are the vaginal rings; these rings are silastic made and allow a continuous release of levonorgestrel or progestins.

Algestone↗

Implications of fertility patterns in the Republic of Korea: a study by computer simulation.

Implications of various fertility patterns in the Republic of Korea (ROK) are studied by computer simulation. These patterns include the factors: age of marriage, sex preference, abortion (induced), contraception and sterilisation. As a result of studying a large number of combinations of these factors, the following tentative conclusions, among others, are reached. If future cohorts of women in the ROK become limiters after two live births and marry according to an observed high level marriage distribution, at least two of the methods of birth control--abortion, contraception, and sterilisation--will have to be used extensively to yield a mean number of live births (MNLB) in these cohorts of the order 2-2. However, if these women marry later than the observed high level marriage distribution and, in addition, accept effective methods of contraception more widely and use them more effectively, then the MNLB experienced by these cohorts could move below 2-5 wihtout the extensive utilisation of abortion and sterilisation.

Abortion, Induced↗

Analysis of G protein alpha subunit mRNA abundance in preimplantation mouse embryos using a rapid, quantitative RT-PCR approach.

We have developed a novel reverse transcription-polymerase chain reaction (RT-PCR)-based approach for systematically quantifying in a single experiment the abundances of many different mRNAs in preimplantation mouse embryos. With this approach, the entire mRNA population from a small number of embryos is amplified while preserving the relative abundance of each mRNA in the cDNA population. The cDNA is analyzed by quantitative hybridization to radiolabeled probes. The approach is very sensitive and provides reliable, quantitative data regarding changes in mRNA abundance. A major advantage of this method is that estimates of mRNA copy number can be obtained and compared between different mRNAs. With this approach, we analyzed the patterns of expression of nine G protein alpha subunit mRNAs (G alpha s, G alpha i, G alpha q, G alpha o, and G alpha 11-15) in oocytes, eggs, and preimplantation embryos from fertilization to the blastocyst stage. Six alpha subunit mRNAs were expressed at significant levels, all of which underwent significant temporal alterations in expression. The mRNAs encoding some alpha subunit types were expressed predominantly in the egg and 1-cell embryo, underwent sharp reductions during the 2-cell stage, and were re-expressed between the 8-cell and blastocyst stages. One alpha subunit mRNA increased in abundance at the early blastocyst stage. The possible significance of these alterations in G protein mRNA abundance to embryonic development is discussed.

Animals↗

Pathophysiological implication of ganglioside GM3 in early mouse embryonic development through apoptosis.

Apoptosis may occur in early embryos where the execution of essential developmental events has failed, and gangliosides, sialic acid-conjugated glycosphingolipids, are proposed to regulate cell differentiation and growth. To evaluate the regulatory roles of ganglioside GM3 in early embryonic development, this study examined its expressional patterns in apoptotic cells during early embryonic development in mice. Pre-implanted embryos were obtained by in vitro fertilization, which were treated at the 4-cell stage with three the apoptosis inducers, actinomycin D, camptothecin and cycloheximide, for 15 h. All three inducers significantly increased the percentage of apoptotic cells, as measured using a TUNEL method, but remarkably reduced the total cell numbers. The numbers of morula and blastocyst stages were significantly decreased by treatment of the embryos with the three apoptosis inducers compared with the control, with a similar result also observed in the number of blastomeres. Staining of early embryos with Hoechst 33342 revealed a significant percentage of apoptotic nuclei. Prominent immunofluorescence microscopy revealed a significant difference in the ganglioside GM3 expression in apoptotic embryos compared with the control, and RT-PCR also demonstrated a dramatic increase in ganglioside GM3 synthase mRNA in the apoptotic embryos. These results suggest that ganglioside GM3 may be pathophysiologically implicated in the regulation of early embryonic development through an apoptotic mechanism.

Animals↗

Morphological and cytogenetic analysis of human giant oocytes and giant embryos.

BACKGROUND: Giant binuclear oocytes occur with considerable frequency in human ovaries, but their ultimate fate remains unknown. We report the morphology, cytogenetics and developmental potential of human giant oocytes from patients undergoing assisted reproductive technologies. METHODS AND RESULTS: A total of 44 giant oocytes was collected from patients aged 22-44 years old, with an overall frequency of 0.3% (44/14 272 oocytes). Giant oocytes were approximately 30% larger in diameter than normal oocytes (mean 200.4 versus 154.7 micro m, P = 0.0001). Two different morphological patterns were observed among giant unfertilized and fertilized oocytes. All unfertilized oocytes appeared to be diploid and contained either one or two metaphase plates (46 or 2 x 23 chromosomes), and one or two polar bodies respectively. Consequently, fertilized giant oocytes exhibited either two or three pronuclei, or two or four polar bodies. Both types of giant zygotes were capable of normal cleavage and development to blastocyst stage. Four giant embryos were analysed by interphase fluorescence in-situ hybridization using probes for chromosomes 9, 22, X and Y, and all appeared chromosomally abnormal with numerical alterations indicative of ploidy change. CONCLUSIONS: Giant oocytes might be a possible source of human digynic triploidy. To avoid undesired miscarriages, giant embryos originated from either two- or three-pronuclear giant zygotes should be excluded from uterine transfers.

Adult↗

[Experiences with levonorgestrel in postcoital contraception].

To examine levonorgestrel as a postcoital contraceptive, 77 women received an oral dose of 0.4 mg per coitus for 1011 cycles and 27 women were administered 0.75 mg per coitus for 226 cycles. In the first dose group seven women became pregnant (Pearl's index 8.3), while two pregnancies resulted in the second group, one of the latter because of faulty drug intake (uncorrected Pearl's index 10.6; corrected Pearl's index 5.3). Menstrual irregularities (chiefly break-through bleedings and oligomenorrhea) were observed in 84.4% and 88.9% of the women, respectively. The number of cycle disorders increased with increasing intake rate and diminished when the drug was applied in excess of six months. The experiments undertaken to test the mechanisms of action indicated an influence on both cervical factor and endometrium, whereas the occurrence of LH-peaks and biphasic basal body temperature patterns suggested the presence of ovulations. Application of levonorgestrel for postcoital contraception failed to be a reliable routine method of hormonal contraception because of insufficient efficacy and considerable menstrual irregularities. The drug should be administered only after unprotected sexual intercourse as might happen, to women with very low frequency of intercourse, or in periods of reduced fertility.

Adolescent↗

Mucus observations in the fertile window: a better predictor of conception than timing of intercourse.

BACKGROUND: Intercourse results in a pregnancy essentially only if it occurs during the 6-day fertile interval ending on the day of ovulation. The strong association between timing of intercourse within this interval and the probability of conception typically is attributed to limited sperm and egg life times. METHODS: A total of 782 women recruited from natural family planning centres in Europe contributed prospective data on 7288 menstrual cycles. Daily records of intercourse, basal body temperature and vaginal discharge of cervical mucus were collected. Probabilities of conception were estimated according to the timing of intercourse relative to ovulation and a 1-4 score of mucus quality. RESULTS: There was a strong increasing trend in the day-specific probabilities of pregnancy with increases in the mucus score. Adjusting for the mucus score, the day-specific probabilities had limited variability across the fertile interval. CONCLUSIONS: Changes in mucus quality across the fertile interval predict the observed pattern in the day-specific probabilities of conception. To maximize the likelihood of conception, intercourse should occur on days with optimal mucus quality, as observed in vaginal discharge, regardless of the exact timing relative to ovulation.

Adult↗