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PubMed1986

Contraceptive practices of female physicians.

To evaluate a potential bias between methods of birth control used and prescribed by physicians, we surveyed sexually active female physicians in regards to their own methods of birth control. When the contraceptive practices of female physicians are compared to those of the general population, no difference in use of various contraceptive methods is found. Among female obstetrician-gynecologists, however, the intrauterine device continues to be a disproportionally popular method of contraception. It is concluded that no gender bias exists in prescribing patterns of contraceptives since contraceptive use in female physicians is identical to that of the general populations.

Adult↗
PubMed2003

Are body fluctuating asymmetry and the ratio of 2nd to 4th digit length reliable predictors of semen quality?

BACKGROUND: Recent attention has been paid to patterns of fluctuating asymmetry (FA) in paired bilateral traits and the extent to which they reflect phenotypic and genetic quality. The FA-fertility hypothesis proposes that FA may be a reliable indicator of ejaculate quality in humans and other animals. The common control by the Hox genes of the differentiation of both the urogenital system and the appendicular skeleton in vertebrates has been proposed as an explanation for the recent finding that FA, and the second to fourth digit ratios (2D:4D) are both associated with semen quality in men. METHODS: A group of 50 men was evaluated for FA, calculated by the sum of three different body FAs, 2D:4D ratios, and seminal parameters of masturbatory semen samples. RESULTS: Composite FA had a significant effect on semen parameters; the 2D:4D ratios did not predict semen quality. CONCLUSIONS: Comparison of our data with previous studies suggests that the putative relationship between semen quality and 2D:4D may have been driven by the inclusion of severely oligozoospermic men within the original subject group. Our sample included men with equally high 2D:4D ratios but who had normal semen. Thus, the 2D:4D ratio may not reliably indicate poor semen quality although FA might.

Adult↗
PubMed1997

Human sperm cytosolic factor triggers Ca2+ oscillations and overcomes activation failure of mammalian oocytes.

Among the possible mechanisms of oocyte activation after sperm penetration, it appears most likely that a protein released by the spermatozoon elicits a calcium elevation in the ooplasm. To further test this idea, cytosolic factors obtained from human spermatozoa by two different methods, freezing-thawing and sonication, were injected into mouse oocytes following which intracellular calcium release was measured. Of a total of 42 mouse oocytes, a pattern of calcium oscillations was observed in nine out of 16 oocytes injected with sonicated fraction, in all of eight oocytes with the frozen-thawed fraction and in none of 18 control oocytes. Injection of the frozen-thawed fraction also produced regular calcium oscillations in all of five in-vitro matured human oocytes. To assess the putative factor's ability to support fertilization, human oocytes that were not activated by prior intracytoplasmic injection of spermatozoa (ICSI) and round spermatids were reinjected with the frozen-thawed sperm fraction. Of 23 human oocytes which remained unfertilized after ICSI, 19 became activated after injection with sperm cytosolic factor; eight showed two pronuclei, three one pronucleus and eight showed three or more pronuclei. Of 11 oocytes unfertilized after prior round spermatid injection, two developed two pronuclei, four developed one pronucleus and two had three or more pronuclei. Cytogenetic analysis by fluorescence in-situ hybridization confirmed the existence of a male pronucleus in eight out of nine such zygotes displaying two or more pronuclei. Thus, human sperm extracts activated mouse and human oocytes after injection, as judged by calcium flux patterns in conjunction with male pronucleus formation.

Animals↗
PubMed1996

Proliferative activity in the different ovarian compartments in cycling rats estimated by the 5-bromodeoxyuridine technique.

The ovary is one of the most active proliferating tissues, and several methods for assessing cellular proliferation have been used to study follicular kinetics during the estrous cycle in the rat. In this study, we have estimated the proliferative activity of the different ovarian tissue compartments by using the 5-bromodeoxyuridine (BrdU) technique. Adult cyclic rats were studied 1 h after pulse-labeling with BrdU. Follicles were divided by size as small (mean diameter < 275 micrograms) or large (mean diameter > or = 275 micrograms). Small follicles were recorded into five classes: 1) primordial follicles (formed by flattened pre-granulosa cells), 2) unilaminar primary follicles (which ranged from having at least one enlarged pre-granulosa cell up to having a complete layer of cuboidal granulosa cells), 3) multilaminar class a (having 1-2 layers of granulosa cells and measuring up to 75 micrograms in diameter), 4) multilaminar class b (76-150 micrograms), and 5) multilaminar class c (151-274 micrograms). Large follicles were recorded into classes 1-5 according to previous classifications [Osman P. J Reprod Fertil 1985; 73:261-270]. Primordial follicles and about 70% of unilaminar primary follicles were unlabeled, whereas all of the remaining classes of small and large follicles were labeled. The labeling index (percentage of BrdU-positive cells) increased in parallel with follicle size but showed a progressive decrease in large preovulatory follicles and a centripetal pattern of cell proliferation. Atretic follicles showed a significantly lower number of labeled cells than healthy follicles of the same size. The number of proliferating cells was not related to follicle size but was inversely correlated with the degree of atresia. Cyclic CL showed intense proliferative activity that was well correlated with functional activity. The number of labeled cells was maximal in metestrus, decreased in the succeeding days, and was very low in proestrus. Proliferating cells seemed to correspond to vascular cells lining blood vessels. These results indicate that the BrdU technique is a useful, accurate, and sensitive method for assessing cellular proliferation in the rat ovary.

Animals↗
PubMed1991

Application of gene amplification by polymerase chain reaction to genetic analysis of molar mitochondrial DNA: the detection of anuclear empty ovum as the cause of complete mole.

To investigate the pathogenesis of complete hydatidiform mole (complete mole), we employed a newly developed gene amplification method by polymerase chain reaction (PCR) for the restriction fragment length polymorphism (RFLPs) analysis of extranuclear DNA (mitochondrial DNA) of complete mole. Whole cellular DNA was extracted from six molar tissues and from peripheral blood mononuclear cells of parents. Two hyperpolymorphic regions of mitochondrial DNA, a 1.5-kb-long fragment and a 1.9-kb-long fragment, were selectively amplified from the extracted DNA by the PCR method. The amplification products amounted to over 10 micrograms after 30 cycles of PCR. The PCR products were digested with endonucleases (HaeIII, HinfI, AluI, and TaqI) and then electrophoresed on agarose gel. The electrophoretic pattern of digested DNA showed that the RFLPs of molar mitochondrial DNA coincided with those of the patient, indicating that the mitochondrial DNA of complete mole was inherited from the ovum. As it has been identified that the intranuclear DNA of complete mole is transmitted only from the spermatozoa, our results verify that complete mole results from the fertilization of an anuclear "empty" ovum with normal sperm at the molecular level.

Base Sequence↗
PubMed2001

Differential gene expression in pre-implantation embryos from mouse oocytes injected with round spermatids or spermatozoa.

BACKGROUND: The use of immature male germ cells to fertilize human oocytes raises several questions. Spermatozoa are normally quiescent, but many genes are transcribed post-meiotically in round spermatids. This creates a novel situation for the oocyte. We have therefore explored the effects on early embryonic development of introducing a fully transcriptionally active round spermatid into the oocyte. METHODS AND RESULTS: Following the micro-injection of spermatozoa or spermatids into mouse oocytes we have analysed the expression, at various times, of six genes in the resulting embryo. Spermatozoa and spermatids produced similar fertilization rates. Hprt was expressed in all embryos at all stages tested. Hsp70.1 was found normally during the 2-cell stage and repressed by the 4-cell stage in embryos from both spermatozoa and round spermatids. However, the amplitude of the signal was greatly reduced in 2-cell embryos from round spermatids. Smcy also showed a disturbed pattern of expression in embryos from round spermatids. Protamine 2, which is normally restricted to the spermatid stage, was expressed following fertilization with round spermatids, but was already repressed at the two pronuclei stage. Ube1Y, which is normally expressed post-meiotically and not during the post-implantatory development, was expressed up to the 2-cell stage in embryos from round spermatids only, and then repressed. Ube1X was also expressed up to the 2-cell stage, but in both embryo types. CONCLUSIONS: We therefore suspect that in embryos fertilized with round spermatids, regulatory mechanisms for inhibiting the inappropriate transcription of male post-meiotically expressed genes are activated following fertilization, permitting the zygotic genome activation to occur, though with some disturbances.

Animals↗
PubMed1997

Analysis of P[gal4] insertion lines of Drosophila melanogaster as a route to identifying genes important in the follicle cells during oogenesis.

We report the analysis of a number of lines of Drosophila melanogaster containing insertions of the yeast gal4 gene. By crossing a UAS-lacZ fusion gene as a reporter into these lines, we analysed the expression patterns of beta-galactosidase during oogenesis. Since there is no expression of GAL4 in the germ-line in these experiments, this is an ideal system for the analysis of expression patterns in sub-sets of follicle cells. These lines provide ideal markers for sets of follicle cells, e.g. anterior or posterior polar cells for studying genetic interactions in oogenesis; however, they can also be used in the same way as conventional enhancer traps to clone nearby genes with similar expression patterns. The advantages of this dual gal4/UAS system over conventional enhancer trapping includes the possibility of GAL4-directed misexpression and antisense expression studies to establish the function of the genes we identified during follicle cell determination and differentiation. These studies could lead to the isolation of homologous genes crucial in mammalian oogenesis. Understanding how the somatic cells and germ cells interact to promote growth and maturation of the mammalian follicle and oocyte could well be crucial for improving the fertility of eggs used for in-vitro fertilization programmes, and could provide methods for assessing the quality of eggs.

Animals↗
PubMed1981

Use of spermatozoa for artificial insemination. I. Fertilizing capacity of fresh and frozen spermatozoa in sows on 36 farms.

A field trial was conducted on 36 farms in the Netherlands to compare the fertilizing capacity of fresh and frozen-thawed boar spermatozoa. Four-hundred and fifty-one sows were artificially inseminated with semen than that had been frozen and thawed according to the Beltsville Method or diluted in Kiev extender and inseminated on the day of collection. Twelve boars of the Dutch Landrace and Dutch Large White breeds were used. Farrowing rates, total number of pigs per litter and number of live pigs per litter were higher (P less than .0001) for sows inseminated with fresh semen than for sows inseminated with frozen-thawed semen (79.1%, 10.6 and 9.9 vs 47.0%, 7.4 and 7.1, respectively). Farrowing rates for sows inseminated with frozen-thawed semen were higher when semen from Dutch Large White boars was used than when semen from Dutch Landrace boars was used (58.6 vs 40.9%); the pattern reversed for insemination with fresh semen (76.5 vs 81%). Boar differences based on farrowing rate ranged from 62 to 92% for fresh semen and from 29 to 72% from frozen semen. There was no inseminator effect or farm effect on farrowing rate. On the basis of these results, frozen semen used for artificial insemination under practical circumstances can be expected to result in a farrowing rate about 30 percentage points lower and a litter size about three pigs smaller than does fresh semen.

Animals↗
PubMed2006

The ORTHO BC-SAT--a satisfaction questionnaire for women using hormonal contraceptives.

OBJECTIVE: To assess the reliability and validity of the ORTHO Birth Control Satisfaction Assessment Tool (ORTHO BC-SAT). DESIGN: 339 women using 1 of 4 hormonal birth control methods (oral contraceptives, transdermal patch, vaginal ring, injections), completed the questionnaire 1-2 times. MATERIALS AND METHODS: The questionnaire was developed based on findings from the literature, focus groups, and interviews. Internal consistency reliability, test-retest reliability, construct validity, and known groups validity were evaluated. RESULTS: Based on variable clustering, 8 domains were identified (Ease of Use/Convenience, Compliance, Lifestyle Impact, Symptom/Side Effect Bother, Menstrual Impact, Future Fertility Concerns, Assurance/Confidence, Overall Satisfaction). Internal consistency reliability was demonstrated with Cronbach's alpha values ranging from 0.70 to 0.89. All multi-item scales reported acceptable test-retest reliability (0.79-0.87). Construct validity was demonstrated by support of a hypothesized pattern of correlations. Known groups validity was confirmed by examining scale scores of women categorized by levels of symptom bother. As expected, women with the least amount of bother reported higher scores on all satisfaction scales than those with higher bother (p < 0.0001), except on Future Fertility Concerns (p = 0.27). CONCLUSION: Our results support the reliability and validity of the ORTHO BC-SAT. It may be used in future studies to evaluate satisfaction among hormonal contraceptive users.

Adult↗
PubMedDate not supplied

In vitro fertilization: increased VLA (very late antigen) integrins and fibronectin after acrosome reaction.

The pattern of detectable adhesion molecules (AM) on human ejaculated spermatozoa used for in vitro fertilization (IVF) of human oocytes was evaluated. The percentage of spermatozoa with the alpha chains 3, 4, 5, and 6 of the integrins (also called very late antigens, VLA alpha 3, 4, 5, 6), the alpha V and the beta 3 chains, as well as the matrix proteins laminin and fibronectin, were determined by flow cytometry using monoclonal antibodies against integrins and matrix proteins before and after the acrosome reaction. This reaction was induced by the low-temperature method according to P. Sanchez, E. Töpfer-Petersen, R. J. Aitken, and W. B. Schill (1991, Andrologia 23:197-203) and monitored by the binding of FITC-conjugated Pisum sativum agglutinin. Twenty-three of 34 investigated semen samples fertilized at least one oocyte (fertile group) and 11 samples did not induce a pronucleus formation or cleavage of an oocyte (infertile group). Significant differences in the classical semen parameters between both semen sample groups could be shown only for the percentage of progressively motile spermatozoa (p = .046, U test). The spermatozoa of the fertile group showed a significantly higher expression of VLA alpha 3 (p = .045) and fibronectin (p = .048). Additionally, after the loss of the acrosome a significantly higher expression of alpha 4, alpha 5, and alpha 6 chains of integrins (p < .05) was detected in the fertile semen group. In contrast, the inducibility of the acrosome reaction differed only to a nonsignificant extent (p = .094). These data suggest that spermatozoa showing a higher expression of the alpha chains of the integrins after the acrosome reaction have a better fertilization rate in vitro.

Acrosome↗
PubMedDate not supplied

Enzymes of human cervical mucus: a comparative study on fertile and infertile mucus and on the endocervical epithelium.

We report a comparative study on protein and enzyme content of human periovulatory cervical mucus and endocervical epithelium. The salient results of our investigation are summarized in the following three points: 1) The method of taking of cervical mucus samples may markedly affect the results of protein and enzyme assays. It is warmly advisable to collect mucus samples from the endocervical canal. 2) Cervical mucus from women diagnosed as having cervical-factor infertility is oligoproteic and lacks some enzyme activities always present in normal cases. 3) Human cervical mucus and endocervical epithelium are very similar as for the enzyme distribution, but show quite different LDH-isoenzyme patterns. In the latter tissue, the H subunits constitute about 77 per cent of the whole LDH, suggesting that this enzyme here is functioning to produce pyruvate rather than lactate.

Adult↗
PubMed1996

Contraceptive patterns among women and men in León, Nicaragua.

The aim was to study the contraceptive patterns among men and women in León, Nicaragua. A questionnaire about sexual, contraceptive, reproductive and socioeconomic issues was directed to 7,789 households including 22% of all women of the municipality aged 15-49 years (n = 10,867). A subsample of 388 men and 413 women aged 15-49 years was drawn at random. Refusals were less than 2%. Private interviews revealed that among fertile women who had been sexually active within the last three months, non-pregnant and wishing to avoid pregnancy, 77% were contracepting. Female sterilization was the most common contraceptive method (39%), followed by intrauterine device (16%). Even though around 60% of women at some time had tried oral contraceptives, only 13% of contraceptors used them currently. The rhythm and interruption methods together constituted only 4%. Condom use was low and mainly occasional. Contraceptive use in sexually active women aged 15-44 years was lower among those having lower education, living in rural areas, and living under poverty conditions. The predominance of female sterilization and the occasional condom use-mainly reported by men-reflects a situation of relative male control over contraception and reproduction. This probably originates from "machista" values where men having many children with different women are considered strong. There was also a significantly higher use of contraceptives among the better-off women and men compared with the extremely poor. The situation of many poor women, in a country with limited contraceptive services, is worrying considering that abortion is illegal and the threat of HIV epidemic is growing. The situation for adolescents is particularly problematic with low experience in contraceptive use.

Adolescent↗
PubMed1999

Studies on renin-2 gene in transgenic rats.

OBJECTIVE: To determine the function of, in vivo, renin and its role in the pathogenesis of hypertension. METHODS: A renin-2 gene restriction map was constructed by endonuclease digestion and Southern blotting hybridization. Transgenic rats were produced via microinjection method. RESULTS: The 24 kb fragments containing mouse full-length ren-2 and its flanking sequence were cleaved by single enzymes (EcoR I, Kpn I and BamH I) and combined enzymes (EcoR I/Kpn I, Kpn I/BamH I and BamH I/EcoR I), respectively. The digests were electrophoresed in 0.8% agarose plates and transferred onto NC membranes. Radioactive 735 bp and 1400 bp probes obtained from half and full-length renin-1 cDNA were used in southern blotting hybridization. According to the electrophoresis and hybridization patterns, a ren-2 restriction map was constructed. 1603 fertilized rat ova after injection with purified 24 kb renin-2 gene were implanted into the oviducts of 81 pseudopregnant recipients in about 20 ova per female rat. 306 progenies were obtained from 50 foster mothers (average of pregnancies was 56.6%). 248 survived pups were identified by PCR analysis and Southern hybridization, and eight positive rats were found to be the transgenic rats (founder, F). All of them carried long fragments (24 kb) of renin-2 gene with normal blood pressure. Preliminary breeding and screening were carried out in the founder. Total survival pups (17.8%) and overall efficiencies (1%) were harvested as the same as those reported in the literatures. A systemic observation and the problems occurred during production of transgenic rats were also described besides the technique procedure used in this study. CONCLUSIONS: Mapping of full-length murine ren-2 can be used in investigation of the structure and function of the gene. The results denoted that the ren-2 transgenic rats were successfully established in this study and the technique used in the production of transgenic rats was proved to be valid in leading to wide-spread application of transgenic technique to many other related researches.

Animals↗
PubMed1989

Oral contraceptives and breast cancer.

A population-based case-control study of oral contraceptive use and breast cancer was carried out among young women (less than 43 years of age) at Group Health Cooperative of Puget Sound, Seattle, Washington. Use of oral contraceptives before first pregnancy did not materially differ between cases or controls. The rate ratio estimate of breast cancer incidence in women who had used oral contraceptives before first pregnancy compared to those who had not was 0.9 (95% CI = 0.4, 2.1). There were no meaningful patterns of association between breast cancer and duration of use or formulation of oral contraceptive used before first pregnancy.

Adult↗
PubMed2003

Scanning of Y-chromosome azoospermia factors loci using real-time polymerase chain reaction and melting curve analysis.

OBJECTIVE: To develop a novel method to scan AZF loci looking for microdeletions. DESIGN: Molecular method development. SETTING: Men undergoing reproductive techniques in a private fertility unit. Molecular methods were performed in a private center for biomedical research. PATIENT(S): : Fifty-eight men divided in two groups depending on seminal analyses. A group of 19 women were also included as positive controls (absence of amplification). INTERVENTION(S): Peripheral blood extraction and DNA purification. MAIN OUTCOME MEASURE(S): Our method is based on real-time polymerase chain reaction (PCR) and melting curve analysis. We performed the screening of 16 selected sequence tagged sites (STS) within AZF loci, and we also calculated the mean, range, and standard deviation for melting temperature patterns and the crossing points values for each STS tested. RESULT(S): We detected one azoospermic patient with several STS deleted within the AZFc region. No deletions were detected in a group of 13 healthy men, and no amplification for any of the STS tested were observed in the positive control group (19 healthy women). CONCLUSION(S): We have developed a novel method based on real-time PCR and melting curve analysis to scan AZF loci looking for microdeletions This method is fast and reliable and permits the scanning of DNA from one patient per hour, minimizing the risk of cross contamination, and false-positive and false-negative results.

Adult↗