Showing papers in "Human Reproduction in 1995"

Polycystic ovary syndrome: the spectrum of the disorder in 1741 patients.

Abstract: The criteria for the diagnosis of the polycystic ovary syndrome (PCOS) have still not been agreed universally. A population of 1741 women with PCOS were studied, all of whom had polycystic ovaries seen by ultrasound scan. The frequency distributions of the serum concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone and prolactin and the body mass index, ovarian volume, uterine cross-sectional area and endometrial thickness were determined and compared with the symptoms and signs of PCOS. Obesity was associated with hirsutism and an elevated serum testosterone concentration and was also correlated with increased rates of infertility and cycle disturbance. The rates of infertility and cycle disturbance also increased with serum LH concentrations > 10 IU/l. A rising serum concentration of testosterone [mean and 95th percentiles 2.6 (1.1-4.8) nmol/l] was associated with an increased risk of hirsutism, infertility and cycle disturbance. The ovarian volume was correlated with serum concentrations of testosterone, LH and the body mass index, which was also correlated with the uterine area. This descriptive data from the largest reported series of women with PCOS enables the development of a management-orientated approach to the syndrome. Women who are overweight can expect an improvement in their symptoms if they lose weight. An elevated concentration of LH (> 10 IU/l) is associated with infertility and treatment should be chosen accordingly. If the serum testosterone concentration is > 4.8 nmol/l, other causes of hyperandrogenism should be excluded.

ATP content of human oocytes and developmental potential and outcome after in-vitro fertilization and embryo transfer.

Abstract: The relationship between the ATP content of mature human oocytes and developmental potential after uterine transfer of sibling embryos was examined in 20 non-male factor in-vitro fertilization (IVF) patients matched for age, fertility history, ovarian stimulation protocol, oocyte quality and number, stage and morphology of embryos at uterine transfer. ATP content was determined for uninseminated and unfertilized oocytes for each cohort, and for embryos that developed after dispermic fertilization or which showed significant fragmentation. The effect of reduced ATP content on meiotic maturation, fertilization and preimplantation development was examined in cultured mouse oocytes treated with uncouplers of mitochondrial oxidative phosphorylation. The results demonstrated that meiotic maturation occurs in both mouse and human oocytes over a wide range of ATP contents, and that the ATP content of normal-appearing, metaphase II human oocytes can differ significantly between cohorts; however, a higher potential for continued embryogenesis and implantation in the human is associated with embryos that develop from cohorts of oocytes with ATP contents > or = 2 pmol/oocyte. The findings are discussed with respect to possible aetiologies and developmental consequences for embryonic development of different oocyte ATP contents, and the extent to which mitochondrial function may determine or influence the continued developmental capacity of embryos which appear normal and developmentally viable at the early cleavage stages.

Weight loss results in significant improvement in pregnancy and ovulation rates in anovulatory obese women

Abstract: Obesity can affect ovulation and the chances of pregnancy. In this prospective study, a weight loss programme was assessed to determine whether it could help infertile overweight anovulatory women to establish ovulation and assist in achieving pregnancy, ideally without further medical intervention. The subjects acted as their own historical controls. They underwent a weekly programme of behavioural change in relation to exercise and diet over 6 months ; those who did not complete the 6 months were treated as the comparison group. Women in the study group lost an average of 6.3 kg, with 12 of the 13 subjects resuming ovulation and 11 becoming pregnant, five of these spontaneously. Fitness, diet and psychometric measurements all improved. Fasting insulin and testosterone concentrations dropped significantly, while sex hormone binding globulin concentrations rose. None of these changes occurred in the comparison group. Thus, weight loss with a resultant improvement in ovulation, pregnancy outcome, self-esteem and endocrine parameters is the first therapeutic option for women who are infertile and overweight.

Pregnancies after testicular sperm extraction and intracytoplasmic sperm injection in non-obstructive azoospermia

Abstract: In this study (May 1 until August 31, 1994) a total of 15 azoospermic patients suffering from testicular failure were treated with a combination of testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI). Spermatozoa were available for ICSI in 13 of the patients. Out of 182 metaphase II injected oocytes, two-pronuclear fertilization was observed in 87 (47.80%); 57 embryos (65.51%) were obtained for either transfer or cryopreservation. Three ongoing pregnancies out of 12 replacements (25%) were established, including one singleton, one twin and one triplet gestation. The ongoing implantation rate was 18% (six fetal hearts out of 32 embryos replaced).

Impact of reactive oxygen species on spermatozoa: a balancing act between beneficial and detrimental effects

Abstract: Reactive oxygen species (ROS) have beneficial or detrimental effects on sperm functions depending on the nature and the concentration of the ROS involved, as well as the moment and the location of exposure. Excessive generation of ROS in semen, mainly by neutrophils but also by abnormal spermatozoa, could be a cause for infertility. Hydrogen peroxide is the primary toxic ROS for human spermatozoa. Low concentrations of this ROS do not affect sperm viability but cause sperm immobilization mostly via depletion of intracellular ATP and the subsequent decrease in the phosphorylation of axonemal proteins. High concentrations of hydrogen peroxide induce lipid peroxidation and result in cell death. On the other hand, the superoxide anion appears to play a major role in the development of hyperactivation and capacitation. The observations that: (i) exogenously generated superoxide anions induce hyperactivation and capacitation; (ii) capacitating spermatozoa themselves produce elevated concentrations of superoxide anion over prolonged periods of time; and (iii) removal of this ROS by superoxide dismutase prevents sperm hyperactivation and capacitation induced by various biological fluids, stress the importance of the superoxide anion in these processes.

The result of intracytoplasmic sperm injection is not related to any of the three basic sperm parameters.

Abstract: High success rates have been reported for the use of intracytoplasmic sperm injection (ICSI) in alleviating essentially andrological infertility. However, neither the relationship between any of the sperm parameters and the result of ICSI nor the minimal sperm requirements for ICSI have been investigated so far. In this paper, our objective was therefore to study the relationship between three basic sperm parameters (total sperm count, sperm motility and morphology) and the outcome of ICSI by retrospective analyses of fertilization, embryo development and pregnancy rates in 966 micro-injection cycles, performed with ejaculated semen. The results showed that there was no important influence from either the type or the extent of sperm impairment on the outcome of ICSI. Even in the most extreme cases of male-factor infertility, where cryptozoospermia or total astheno- or total teratozoospermia was diagnosed in the initial semen sample, high fertilization and pregnancy rates were obtained by ICSI. Only one condition had a strongly negative influence on the result of ICSI: where an immotile (presumably dead) spermatozoon was injected into the oocyte. Thus the only ultimate criterion for successful ICSI is the presence of at least one living spermatozoon per oocyte in the pellet of the treated semen sample used for micro-injection.

High fertilization and pregnancy rate after intracytoplasmic sperm injection with spermatozoa obtained from testicle biopsy

Abstract: In cases requiring microsurgical epididymal sperm aspiration (MESA) for congenital absence of the vas deferens (CAVD) or irreparable obstructive azoospermia, often no spermatozoa can be retrieved from the epididymis, or there may even be no epididymis present. We wished to see whether testicular biopsy with testicular sperm extraction (TESE) in such cases could yield spermatozoa that would result in successful fertilization and pregnancy (despite the absence of epididymal spermatozoa) using intracytoplasmic sperm injection (ICSI). In the same setting during the same 2-week period, 28 patients with CAVD or irreparable obstruction were treated; 16 consecutive fresh MESA-ICSI cycles and 12 cycles which required testicular biopsy with testicular sperm extraction (TESE-ICSI) were performed. Normal two-pronuclear fertilization rates were similar in both groups: 45% for epididymal spermatozoa and 46% for testicular biopsy-extracted spermatozoa. Cleavage rates were also similar (68% for epididymal and 65% for testicular spermatozoa). The ongoing pregnancy rates in this series were 50 and 43% respectively. We conclude that epididymal spermatozoa and testicular spermatozoa yield similar fertilization, cleavage and ongoing pregnancy rates using ICSI. When epididymal spermatozoa cannot be retrieved, a testicular biopsy can be performed and the few barely motile spermatozoa thus obtained can be used for ICSI. It appears that all cases of obstructive azoospermia can now be successfully treated.

Embryo score to predict implantation after in-vitro fertilization: based on 957 single embryo transfers.

Abstract: The purpose of this study was to devise an embryo score to predict the likelihood of successful implantation after in-vitro fertilization (IVF). Unlike most studies dealing with the influence of embryo stage and morphology on pregnancy, our study was based on single rather than multiple embryo transfers. A total of 957 single embryo transfers were carried out. No delivery was obtained after any of the 99 transfers using 1-cell embryos or embryos obtained after delayed fertilization. In the remaining 858 transfers, the embryos had cleaved. Higher pregnancy rates were obtained with embryos displaying no irregular cells (11.7 versus 6.9%; P 38 years (8.2 versus 11.4%; P < 0.05), even though embryo quality was similar regardless of age. Single embryo transfer allowed us to define a simple and useful embryo score to choose the best embryo for transfer to optimize IVF and embryo transfer outcome. The use of this embryo score could decrease multiple pregnancies after multiple embryo transfers.

Clinical evidence for a detrimental effect on uterine receptivity of high serum oestradiol concentrations in high and normal responder patients.

Abstract: This study was undertaken to investigate an empirical observation that 'high responder patients have poorer in-vitro fertilization (IVF) outcome than normal responder patients'. The aim of our study was to analyse the effect of high serum oestradiol and progesterone concentrations at the day of human chorionic gonadotrophin (HCG) administration on endometrial receptivity and oocyte-embryo quality in high and normal responder patients. The IVF patients were divided into two groups: 59 high responder patients who voluntarily donated some of their oocytes, and a control group consisting of 105 normal responder patients. Both groups were compared in terms of the number and quality of oocytes retrieved, embryos transferred, fertilization, implantation and gestation rates, serum oestradiol and progesterone concentrations and the oestradiol:progesterone ratio on the day of HCG injection. To ascertain oocyte-embryo quality, a second control group of 96 women undergoing oocyte donation (receiving oocytes from high responder patients) was considered. To assess the impact of steroid concentrations on endometrial receptivity, high responder patients were divided into two subgroups according to oestradiol concentration, above or below the minimal oestradiol and progesterone concentrations (mean--SD) in this group. The normal responder patients were divided into two subgroups according to oestradiol concentration, above or below the maximal oestradiol and progesterone concentrations (mean+SD) in this group. To assess further the relevance of oestradiol concentration on endometrial receptivity, patients were divided into different subgroups according to increasing oestradiol concentration, regardless of whether they were high or normal responders. High responder patients had significantly decreased implantation and pregnancy rates per cycle compared with normal responder patients (33.3 versus 16.3 and 11.1 versus 5.4% respectively; P 1700 pg/ml compared with those having oestradiol concentrations 2200 pg/ml compared with those having oestradiol concentrations 2500 pg/ml compared with patients having lower oestradiol concentrations. Our clinical results demonstrate that high serum oestradiol concentrations on the day of HCG injection in high and normal responder patients, regardless of the number of oocytes retrieved and the serum progesterone concentration, are detrimental to uterine receptivity without affecting embryo quality.

Prevention of osmotic injury to human spermatozoa during addition and removal of glycerol.

Abstract: Use of a cryoprotective agent is indispensable to prevent injury to human spermatozoa during the cryopreservation process. However, addition of cryoprotective agents to spermatozoa before cooling and their removal after warming may create severe osmotic stress for the cells, resulting in injury. The objective of this study was to test the hypothesis that the degree (or magnitude) of human sperm volume excursion can be used as an independent indicator to evaluate and predict possible osmotic injury to spermatozoa during the addition and removal of cryoprotective agents. Glycerol was used as a model cryoprotective agent in the present study. To test this hypothesis, first the tolerance limits of spermatozoa to swelling in hypo-osmotic solutions (iso-osmotic medium diluted with water) and to shrinkage in hyperosmotic solutions (iso-osmotic medium with sucrose) were determined. Sperm plasma membrane integrity was measured by fluorescent staining, and sperm motility was assessed by computer-assisted semen analysis before, during and after the anisosomotic exposure. The result indicate firstly that motility was much more sensitive to anisosmotic conditions than membrane integrity, and secondly that motility was substantially more sensitive to hypotonic than to hypertonic conditions. Based on the experimental data, osmotic injury as a function of sperm volume excursion (swelling or shrinking) was determined. The second step, using these sperm volume excursion limits and previously measured glycerol and water permeability coefficients of human spermatozoa, was to predict, by computer simulation, the cell osmotic injury caused by different procedures for the addition and removal of glycerol. The predicted sperm injury was confirmed by experiment. Based on this study, an analytical methodology has been developed for predicting optimal protocols to reduce osmotic injury associated with the addition and removal of hypertonic concentrations of glycerol in human spermatozoa.

Intracytoplasmic sperm injection in the mouse

Abstract: Intracytoplasmic sperm injection (ICSI) into mouse oocytes involves a very low survival rate. This study was designed to determine why ICSI frequently fails in mice. Metaphase II oocytes were obtained from superovulated 4-6 week old F1 hybrid mice. Spermatozoa were retrieved from the epididymis of 12-14 week old F1 hybrid mice. The spiked microinjection pipette used to inject a spermatozoon into the ooplasm had outer and inner diameters of 10 and 8 microns respectively. The oocytes used in the first part of the study were not activated (group 1). Some oocytes were incubated with calcium ionophore for 5 min (group 2). The injected oocytes were evaluated 6, 20, 48 and 72 h after injection. A total of 143 eggs in each group underwent ICSI. In group 1, sperm heads escaped into the perivitelline space. In all, 63 (47%) of the remaining oocytes were damaged during the injection or had degenerated by the first evaluation. The survival rate was 53%, but fertilization did not occur. In group 2, 31 oocytes (22%) were damaged during microinjection or soon degenerated. Two oocytes underwent accidental subzonal insemination. Six oocytes were fertilized (4.2%) among the 78% of survivors. After injection, the sperm tail was found in the cytoplasm (27 and 31% in groups 1 and 2 respectively), the perivitelline space (45% in both groups) or protruding through the zona pellucida (28 and 23% respectively). More oocytes degenerated when the tail remained in the cytoplasm, i.e. 78% in group 1 and 36% in group 2.

Blastocyst development and birth after in-vitro maturation of human primary oocytes, intracytoplasmic sperm injection and assisted hatching

Abstract: Immature oocyte recovery followed by in-vitro oocyte maturation and in-vitro fertilization is a promising new technology for the treatment of human infertility. The technology is attractive to potential oocyte donors and infertile couples because of its reduced treatment intervention. Immature oocytes were recovered by ultrasound-guided transvaginal follicular aspiration. Oocytes were matured in vitro for 36-48 h followed by intracytoplasmic sperm injection (ICSI). Embryos were cultured in vitro for 3 or 5 days before replacement. Assisted hatching was performed on a day 5 blastocyst stage embryo. Embryo and uterine synchrony were potentially enhanced by luteinization of the dominant follicle at the time of immature oocyte recovery. Mature oocyte and embryo production from immature oocyte recovery were similar to the previous IVF results of the patients. A blastocyst stage embryo, produced as a result of in-vitro maturation, ICSI, in-vitro culture and assisted hatching, resulted in the birth of a healthy baby girl at 39 weeks of gestation.

Molecular aspects of implantation: The signals and molecular pathways involved in implantation, a symbiotic interaction between blastocyst and endometrium involving adhesion and tissue invasion

Abstract: Implantation is a complex process requiring the interaction of the blastocyst, and subsequently the developing embryo with the endometrium. Initially, the detailed cellular interactions implicated in this process were defined. More recently, many signals and molecular pathways are recognized that induce, or regulate the complex series of interactions required for implantation. In this review, the cellular and molecular interactions that take place during implantation are discussed.

High prospective fetal loss rate in untreated pregnancies of women with recurrent miscarriage and antiphospholipid antibodies

Abstract: Antiphospholipid antibodies (APA), lupus anticoagulant (LA) and/or anticardiolipin antibodies (ACA), are associated with thrombosis and recurrent miscarriage. We studied the outcome of 20 pregnancies in women (median age 32 years; range 23-41) with APA (14 LA positive; three immunoglobulin (Ig) G ACA positive; two IgM ACA positive and one LA and IgG ACA positive) and history of recurrent miscarriage (median 4; range 3-11) who declined pharmacological treatment in their next pregnancy. Comparison was made with a cohort of 100 consecutive women (median age 33 years; range 23-44) with recurrent miscarriage (median 4; range 3-10), in whom no underlying cause to account for their pregnancy losses was found. Of the 20 women with APA, 18 (90%) miscarried compared to 34 of the 100 women (34%) with normal investigations (P < 0.001). The majority (94%) of miscarriages in women with APA occurred in the first trimester. Fetal heart activity was seen prior to fetal death in 86% of women with APA compared to 43% of women with normal investigations (P < 0.01). The first trimester loss of embryonic pregnancies is the most common type of miscarriage in women with APA. This may be a result of defective implantation and subsequent placentation.

Cell adhesion molecules on the oocyte and preimplantation human embryo.

Abstract: The presence of cell adhesion molecules on human oocytes, early embryos, and pre-hatched blastocysts was examined by indirect immunofluorescence and compared to the distribution found on first trimester villous placenta with the same antibodies. Six integrin subunits (alpha 3, alpha V, beta 1, beta 3, beta 4, beta 5) were observed consistently throughout preimplantation development. Evidence was also obtained for the presence of integrin subunits alpha 2, alpha 4, alpha L, beta 2, and beta 7 on a small number of oocytes. A more restricted developmental analysis of E-cadherin, ICAM-1, NCAM, and VCAM-1 demonstrated that these cell adhesion molecules are also present on oocytes and early embryos. L-selectin was detected on oocytes but was not found on 8-cell embryos. The oocyte and early blastomeres have complex surfaces in which the integrin and CAM families are represented.

The use of epididymal and testicular spermatozoa for intracytoplasmic sperm injection : the genetic implications for male infertility.

Abstract: The results and rationale of using testicular and epididymal spermatozoa with intracytoplasmic sperm injection (ICSI) for severe cases of male infertility are reviewed. A total of 72 consecutive microsurgical epididymal sperm aspiration (MESA) cases were performed for congenital absence of the vas (CAV) and for irreparable obstructive azoospermia. ICSI was used to obtain normal embryos for transfer and fertilization in 90% of the cases. The overall fertilization rate was 46% with a normal cleavage rate of 68%. The pregnancy and delivery rates per transfer were 58 and 37% respectively. The delivery rate per cycle was 33%. In many cases, no epididymal spermatozoa were available and so testicular sperm extraction (TESE) was used for sperm retrieval. The transfer rate was lower with TESE (84 versus 96%) and the spermatozoa could not be frozen and saved for use in future cycles. However, there was little difference in pregnancy rates using epidiymal or testicular spermatozoa. The results were not affected by whether the obstruction was caused by CAV or failed vasoepididymostomy. Both fresh and frozen spermatozoa gave similar results ; the only significant factor appeared to be the age of the female. Because of the consistently good results obtained using epididymal sperm with ICSI when compared with conventional IVF, and the similarly good results with testicular tissue spermatozoa, ICSI is mandatory for all future MESA patients. All CAV patients and their partners should be offered genetic screening for cystic fibrosis ; hence pre-implantation embryo diagnosis should be available in any full service MESA programme. It is now clear that even with non-obstructive azoospermia, e.g. Sertoli-cell only, or maturation arrest, there are usually some small foci of spermatogenesis which allow TESE with ICSI to be carried out. This means that even in men with azoospermia due to absence of spermatogenesis or to a block in meiosis, there are usually a few spermatozoa available in the testes that are adequate for successful ICSI. Finally, it is likely that some forms of severe male factor infertility are genetically transmitted and although ICSI offspring have been shown to be completely normal, it is possible that the sons of these infertile couples will also require ICSI when they grow up and wish to have a family.

Analysis of sperm movement in relation to the oxidative stress created by leukocytes in washed sperm preparations and seminal plasma.

Abstract: The addition of luminol to unprocessed semen samples resulted in the generation of chemiluminescent signals, the intensity of which was highly correlated with the level of leukocyte contamination. Despite the spontaneous oxidant-generating capacity of seminal leukocytes, no correlations were observed between leukocyte contamination and the fertility status of the subjects or any aspect of the semen profile, including the motility of the spermatozoa or their performance in a hyaluronate penetration assay. Luminol-dependent chemiluminescence and leukocyte contamination were also correlated in washed sperm suspensions prepared either by repeated centrifugation or on discontinuous Percoll gradients. However, in such sperm suspensions, the spontaneous generation of oxidants by contaminating leukocytes (> 2 x 10(4) leukocytes/ml) was invariably associated with a decreased capacity for movement. Moreover, causative associations between leukocyte contamination, reactive oxygen species generation, lipid peroxidation and impaired sperm motility were revealed by experiments involving the selective addition or removal of activated leukocytes. From these observations we can conclude that low concentrations of leukocytes are a common feature of the human ejaculate and can impair sperm function, particularly in the absence of seminal plasma. These findings have implications for our understanding of the importance of leukocytospermia in defining the fertility of human spermatozoa in vivo and in vitro.

Follicular development in cryopreserved marmoset ovarian tissue after transplantation.

Abstract: Pieces of marmoset ovary were frozen by slow cooling in 1.5 M dimethylsulphoxide. The follicles in fresh and frozen tissue were counted and examined for morphological appearance in stained serial sections. The proportion of normal follicles was similar in fresh tissue and frozen tissue examined immediately after thawing. Follicles at all stages of folliculogenesis up to the small antral stage survived freezing and thawing. Fresh and frozen tissue was transplanted underneath the kidney capsules of ovariectomized immunodeficient mice. The establishment of grafts was similar, and oestrogenic activity (cornification of the vaginal epithelium) was observed in the recipients 20 and 16 days after transplantation of fresh and frozen grafts respectively. The total number of follicles and the proportion of normal follicles were similar in fresh and frozen grafts. Grafts of frozen tissue recovered between 7 and 15 days after transfer contained follicles up to the small antral stage of development. Grafts recovered between 21 and 32 days contained follicles at all stages of folliculogenesis, including large antral follicles (1-2 mm diameter). Our results suggest that freezing and thawing do not substantially damage marmoset ovarian tissue, and the cryopreserved tissue retains its ability to support the development of large antral follicles.

Update on treatment of varicocele: counselling as effective as occlusion of the vena spermatica.

Abstract: This prospective randomized study was performed in order to investigate the effects of interventive treatment or counselling on pregnancy rates in infertile couples in whose male partners a varicocele was diagnosed. The present report extends a previous study using the same design. A total of 125 couples were included in the current study while the previous report comprised 95 couples. Couples fulfilling the inclusion criteria were allocated randomly either to interventive treatment (surgical ligation or angiographic embolization of the spermatic vein) (n = 62) or to counselling as the sole treatment (n = 63). Couples were followed over the subsequent 12 months and seen at 3-monthly intervals. At the end of the 12 month period pregnancy rates, as the main outcome measure, were 29% in the group given interventive treatment and 25.4% in the counselled group and were not significantly different. The only significant difference found, regardless of treatment modality, was the wives' age at admittance: the 34 wives achieving a pregnancy were 28.8 +/- 0.6 years (mean +/- SE) old while the 91 non-pregnant wives were 31.2 +/- 0.3 years old (P < 0.05). The study suggests that regular counselling of the infertile couples is as effective as interventive treatment of varicoceles in achieving pregnancies.

A mathematical model of follicle dynamics in the human ovary.

Abstract: A mathematical model has been developed to describe the rates of growth and death of follicles in human ovaries between 19 and 50 years of age. It was based on the numbers of follicles at three successive stages of development, which were obtained by counting follicles in histological sections of ovaries from 52 normal women. The model indicated that follicle dynamics were age dependent, with a transition at 38 years of age when the rate of follicle disappearance increased. The rates of follicle growth increased at successive stages but did not change with age. The annual egress from stage III (consisting of follicles with two or more granulosa cell layers) was affected by the declining numbers of small follicles, and corresponded to 31, nine and one follicles per day at 29-30, 39-40 and 49-50 years of age respectively. The rate of death at stage I (representing small, resting follicles) was the only parameter which varied significantly with age: no evidence of significant atresia was found for this stage in ovaries < or = 38 years old, but there was significant death above this age. As a consequence, only 40% of follicles leaving stage I reached stage III in older ovaries and just 1500 follicles in toto remained at 50 years of age from the 300,000 present at 19 years. This high death rate of small follicles appears to be responsible for advancing the timing of ovarian failure, and therefore of menopause, to midlife in our species.

Andrology: Effects of nitric oxide on human spermatozoa: evidence that nitric oxide decreases sperm motility and induces sperm toxicity

Abstract: Endogenous nitric oxide (NO) is an important functional mediator in several physiological systems, including the reproductive system. However, when generated in excessive amounts for long periods, mainly during immunological reactions, NO is cytotoxic and cytostatic for invading microbes, as well as for the cells generating it and the tissues present around it. Since infertility associated with urogenital tract infection in males and females is also accompanied by reduced sperm motility and viability, it is possible that reduced fertility in these patients is due to NO-induced sperm toxicity. We therefore evaluated the direct effects of NO, chemically derived from S-nitroso-N-acetylpenicillamine (SNAP, 0.012-0.6 mM) and sodium nitroprusside (SNP, 0.25-2.5 mM), on the motility and viability of human spermatozoa. Furthermore, we tested whether inhibition of NO synthesis prevents sperm motility and viability by incubating washed total cells present in the semen (spermatozoa, round cells) with N-nitro-L-arginine-methyl-ester (L-NAME), a NO synthesis inhibitor. Treatment of purified spermatozoa with SNAP or SNP decreased forward progressive sperm motility and straight line velocity, and also increased the percentage of immotile spermatozoa in a concentration-dependent manner. Furthermore, the percentage of immotile spermatozoa positively correlated with the percentage of dead spermatozoa. In contrast to freshly prepared SNAP, SNAP preincubated for 48 h had no effect on the motility and viability of the spermatozoa. Furthermore, as compared to untreated controls, a significantly higher percentage of forward progressive sperm motility as well as viability (P < 0.05) was maintained in washed semen incubated with L-NAME (0.15 mM).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of uterine leiomyomata on the results of in-vitro fertilization treatment

Abstract: The effect of uterine leiomyoma on infertility is subject to controversy. Two main mechanisms associating leiomyomata and infertility have been suggested: the obstruction of gamete transport and impaired implantation. In-vitro fertilization (IVF) is a unique setting for examining the effect of leiomyomata on the implantation rate. This study was designed to determine the impact of uterine leiomyomata on the results of IVF. In a retrospective analysis of IVF cycles, 46 women with documented uterine leiomyoma were compared with a control group with mechanical infertility. The implantation rate and pregnancy outcome in relation to the leiomyoma were observed. Implantation (22.1%/transfer, 6.8%/embryo) and abortion rates (36%) in the study group were similar to the results in the control group with pure mechanical factor. An analysis of IVF results according to the hysteroscopic pretreatment findings revealed that impaired implantation is associated with leiomyoma only where uterine intracavitary abnormalities co-exist. We conclude that implantation rate and pregnancy outcome are impaired in women with uterine leiomyomata only when they cause deformation of the uterine cavity. In patients with leiomyomata associated with an abnormal uterine cavity, surgical treatment should be considered prior to IVF because of the reduced implantation rate.

Uterine pinopodes as markers of the 'nidation window' in cycling women receiving exogenous oestradiol and progesterone.

Abstract: In 14 cycling women participating in an in-vitro fertilization (IVF) donation programme, we examined the timing of the 'nidation window' using as a stage-specific 'marker' the presence of fully developed pinopodes on the apical surface of the luminal uterine epithelium. Each woman received exogenous oestradiol from the second day of their cycle and progesterone starting on day 8 or day 15 of the oestrogenic treatment. The women underwent two biopsies during the same artificial cycle, on either days 6 and 9 or days 8 and 10 of the progesterone treatment. All patients to whom oestradiol was administered for 7 days prior to progesterone administration (n = 9), and two of the five treated with oestradiol for 14 days prior to the addition of progesterone, showed uterine pinopodes in either one or both biopsies. When present on a given day, pinopodes were at the same stage, developing, fully developed or regressing, showing that their total lifespan did not exceed 48 h. Fully developed pinopodes existed for 1 day only which may correspond to the short period of optimal endometrial receptivity observed in animal models. The timing of the presence of fully developed pinopodes varied from patient to patient, but these individual differences were not correlated with progesterone and oestradiol plasma concentrations. The brief duration of the nidation window and the observed individual variations in its timing suggest that the detection of uterine pinopodes could be a valuable tool for the prediction, on an individual basis, of the optimal date for successful egg replacement in IVF patients.

Antiphospholipid antibodies and beta 2-glycoprotein-I in 500 women with recurrent miscarriage: results of a comprehensive screening approach.

Abstract: Five hundred consecutive women (median age 33 years; range 19-45) with a history of recurrent miscarriage (median 4; range 3-16) were screened for the presence of antiphospholipid antibodies (APA)-lupus anticoagulant (LA) and/or anticardiolipin antibodies (ACA). The prevalence of persistently positive tests for LA was 9.6% and for immunoglobulin G (IgG) and immunoglobulin M (IgM) ACA was 3.3 and 2.2% respectively. Only seven women (1.4%) were LA and ACA positive. Repeat testing, after an interval of at least 8 weeks, demonstrated that only 65.7% of LA positive, 36.6% IgG ACA positive and 36.0% IgM ACA positive women on initial testing had a second positive test result. The dilute Russell's viper venom time detected the LA significantly more often than either the activated partial thromboplastin time or the kaolin clotting time (P < 0.001). There was no difference in the gestation of previous miscarriages between APA positive and APA negative women. There was no difference in the plasma beta 2-glycoprotein-I concentrations between APA positive and APA negative women with miscarriages and normal women. All women with a history of recurrent miscarriage should be tested for the presence of both LA and ACA. A second confirmatory test should be performed in those with an initial positive test result.

A prospective, randomized, assessor-blind, multicentre study comparing recombinant and urinary follicle stimulating hormone (Puregon versus Metrodin) in in-vitro fertilization

Abstract: Urinary follicle stimulating hormone (FSH) is being used for the treatment of human infertility. Recently, FSH manufactured by means of recombinant DNA technology with a much higher purity (>99%) has become available. A prospective, randomized, assessor-blind, multicentre (n = 18) study was conducted in infertile women undergoing in-vitro fertilization comparing recombinant FSH (Org 32489, Puregon®) and urinary FSH (Metrodin®). Eligible subjects were randomized (recombinant versus urinary FSH = 3 :2) and pretreated with buserelin for pituitary suppression. FSH was given until three or more follicles with a diameter of at least 17 mm were seen. After oocyte retrieval, fertilization routines were applied according to local procedures. No more than three embryos were replaced. In all, 585 subjects received recombinant FSH and 396 urinary FSH. Significantly more oocytes were retrieved after recombinant FSH treatment (mean adjusted for centre 10.84 versus 8.95, P < 0.0001). Ongoing pregnancy rates per attempt and transfer in the recombinant FSH group were 22.17 and 25.97% respectively, and in the urinary FSH group, 18.22 and 22.02% respectively (not significant). Ongoing pregnancy rates including pregnancies resulting from frozen-thawed embryo cycles were 25.7% for recombinant and 20.4% for urinary FSH (P = 0.05). Compared to urinary FSH, the total dose of FSH was significantly lower with recombinant FSH (2138 versus 2385 IU, P < 0.0001) in a significantly shorter treatment period (10.7 versus 11.3 days, P < 0.0001). No clinically relevant differences between recombinant and urinary FSH were seen with respect to safety variables. It is concluded that recombinant FSH (Puregon) is more effective than urinary FSH in inducing multifollicular development and achieving an ongoing pregnancy.

Endometrial thickness appears to be a significant factor in embryo implantation in in-vitro fertilization

Abstract: To evaluate the role of endometrial thickness and pattern in in-vitro fertilization (IVF), these parameters were prospectively measured in 516 cycles of IVF with embryo transfer at our clinic. Pregnancy and embryo implantation rates were assessed for each mm of endometrial thickness and for each of three endometrial patterns. Embryo implantation, clinical and ongoing pregnancy rates were significantly higher in the patients with an endometrial thickness > 9 mm (24.4, 48.6 and 42.2% respectively) compared with those of 9 mm as well as ring and intermediate endometrial patterns denoted a more favourable prognosis for pregnancy in IVF but thinner endometrium and those exhibiting a solid configuration had an acceptable pregnancy outcome.

Analysis of the incidence and risk factors associated with ectopic pregnancy following in-vitro fertilization and embryo transfer

Abstract: Ectopic pregnancy is a well known complication of in-vitro fertilization (IVF) and embryo transfer. From March 1983 to December 1993, 3000 clinical pregnancies were achieved at Bourn Hall Clinic, including 135 ectopic pregnancies (4.5%). Of these ectopics 20 were heterotopic, eight ovarian, six bilateral tubal and the remainder were singleton tubal pregnancies. The main risk factor identified in the series was a history of pelvic inflammatory disease (P < 0.001). The data also showed that ectopic pregnancy is at present more prevalent among patients in whom tubal damage is the reason for treatment. There was slight statistical evidence (P = 0.05) that patients having ectopic pregnancies received a higher volume of culture medium than those having normal deliveries. There was also an apparent trend (P = 0.07, not significant) that high progesterone/oestradiol ratio on the day of embryo transfer was associated with ectopic pregnancy. There was no statistical evidence of association between ectopic pregnancy and a history of ectopic pregnancy, abortion, still birth, termination of pregnancy, neonatal death, tubal surgery, ovarian stimulation protocol, plasma concentration of oestradiol, luteinizing hormone and progesterone, number of oocytes retrieved, number or quality of embryos transferred, administration of general anaesthesia for embryo transfer, and the number of patent Fallopian tubes. Awareness of the risk factors associated with ectopic pregnancy plays an important part in the early diagnosis of this potentially fatal condition.

Metabolic regulation of androgen production by human thecal cells in vitro

Abstract: The association between hyperinsulinaemia and hyperandrogenism in many women with polycystic ovarian syndrome (PCOS) implies roles for insulin and insulin-like growth factors (IGFs) in the regulation of ovarian androgen production. The aim of the present study was to compare the abilities of insulin, IGF-I and IGF-II to stimulate androgen production by human thecal cells in vitro. Serum-free monolayer cell cultures were established from the ovaries of euandrogenic women undergoing hysterectomy with oophorectomy for non-ovarian indications. Androgen (androstenedione) production was determined after 4 days of culture in the presence of insulin or either of the IGFs (10-100 ng/ml), with and without a maximal stimulatory dose of luteinizing hormone (LH; 10 ng/ml). Interactions with inhibin (30 ng/ml), a putative paracrine regulator of ovarian androgen synthesis, were also tested. The three metabolic hormones exerted similar dose-related effects on androgen production (ED50 < or = 10 ng/ml), which were augmented 2- to 3-fold in the presence of LH and further increased several-fold by the additional presence of inhibin. No treatment with insulin or either IGF stimulated thecal cell growth, but all treatments caused striking morphological changes consistent with enhanced steroidogenesis. These results reveal potent regulatory effects of metabolic hormones on human thecal androgen synthesis, which imply (i) 'progonadotrophic' roles for insulin and IGF-I in regulating normal ovarian androgen production, (ii) a role for insulin in the aetiology of hyperandrogenism (both with and without hyperinsulinism) in PCOS and (iii) paracrine roles for granulosa-derived IGF-II and inhibin in regulating ovarian androgen production.

Rectovaginal septum, endometriosis or adenomyosis: laparoscopic management in a series of 231 patients.

Abstract: A series of 231 cases of deep-infiltrating endometriosis of the rectovaginal septum is presented. Laparoscopic procedures with excision of deep fibrotic endometriotic nodules were performed in all cases. In three cases, the bowel lumen was entered. No other peri-operative complications were observed. Three cases of urinary retention were reported. The nodule resection resulted in considerable pain relief. Histologically, the rectovaginal nodule was similar to an adenomyoma. Indeed it was a circumscribed nodular aggregate of smooth muscle and endometrial glands. This form of disease must be considered as a specific disease which originates from the Mullerian rests present in the rectovaginal septum and we suggest that it be called 'rectovaginal adenomyosis'.

Morphology and biochemistry of in-vitro produced bovine embryos: implications for their cryopreservation

Abstract: Examination of some ultrastructural and physiological characteristics of in-vitro produced bovine embryos may help to explain why such embryos are more sensitive to freezing than their in-vivo derived counterparts. Improvement of embryo survival after freezing can be achieved by changing the conditions of their culture, selection of embryos based on the kinetics of their development, and changing "standard' freezing procedures. Cryopreservation of embryos by vitrification, in particular, seems to yield higher survival than conventional slow freezing. Further development of protocols requires additional embryo transfer studies to ensure that the ability of thawed embryos to develop normally in vivo correlates strongly with in-vitro survival assays.