Tyrosine kinase B receptor and its activated neurotrophins in ovaries from human fetuses and adults.

The signals initiating the growth of primordial follicles are unknown. Growth factors such as neurotrophin 4/5 (NT-4/5) and brain-derived neurotrophic factor (BDNF) may play a role in this process. To investigate the expression of NT-4/5 and BDNF and their receptor tyrosine kinase B (TrkB) in the early developing follicles, we fixed and froze 12 ovarian samples from adolescents/adults and 31 ovaries from human fetuses. The fixed samples were prepared for immunohistochemical staining for NT-4/5, BDNF and the TrkB receptor. Total RNA was extracted from the frozen ovarian samples, and the expression of NT-4/5, BDNF and the TrkB receptor (full length and two truncated isoforms) was investigated by RT-PCR. Products were resolved by 1% agarose gel electrophoresis and image analysis. Immunohistochemical staining revealed the expression of NT-4/5 and BDNF mainly in oocytes and, in a minority of samples, also in the granulosa cells (GCs); TrkB receptor was identified in oocytes and GCs. Transcripts of NT-4/5, BDNF and all forms of TrkB receptor were identified in the samples. To elucidate whether indeed NT-4/5 and BDNF are involved in growth initiation of human primordial follicles, they should be added to the culture medium.

The pharmacologic approach to the prevention of preeclampsia: from antiplatelet, antithrombosis and antioxidant therapy to anticonvulsants.

Preeclampsia has been suggested to be a two-stage disorder of an alteration in placental perfusion (stage 1) leading to generalized vascular endothelial damage (stage 2). Because the mechanism linking the two stages remains unclear, effective primary prevention is still impossible. However, advances made in our understanding of the pathophysiology of preeclampsia have paved the way for secondary and tertiary prevention approaches. Platelets are known to be activated in early pregnancy. They also play a pivotal role in the process of inflammation, as demonstrated by the finding that CD40 ligand is shed from activated platelets to directly initiate inflammation of the vessel wall. According to the Cochrane Library Update summarizing data from over 30,000 women, secondary prevention with antiplatelet drugs is associated with a 19% decrease in the risk of preeclampsia. Additional randomized controlled trials are needed to establish the association between preeclampsia and thrombophilia. The effect of the antithrombotic agent heparin on pregnancy outcome in preeclampsia and its potential preventive action in high-risk patients need to be elucidated. One of the several hypotheses of the pathogenesis of preeclampsia focuses on the oxidative stress caused by the imbalance in prooxidant and antioxidant forces. Preliminary findings on vitamin E and vitamin C supplementation in preeclamptic women are encouraging, and suggest a rationale for larger clinical trials. Although there is currently no explanation for the positive effect of magnesium sulfate on eclamptic seizures, studies have provided enough evidence to encourage its worldwide use as the primary anticonvulsant of choice in the tertiary prevention of maternal and perinatal death in severe preeclampsia/eclampsia. In conclusion, secondary and tertiary prevention of preeclampsia is possible when targeted at reducing maternal and neonatal morbidity and mortality.

Presence of NGF and its receptors in ovaries from human fetuses and adults.

The ability to mature human primordial follicles in vitro would assist fertility restoration. However, the signals initiating growth of primordial follicles are unknown. Growth factors such as nerve growth factor (NGF) may play a role in this process. To investigate the expression of NGF and its receptors, p75 and TrkA, in early developing follicles (mostly primordial, primary and secondary follicles), ten ovarian samples from adolescents/adults aged 13-39 and 33 ovaries from human fetuses aged 19-33 gestational weeks (GW) were obtained and immediately fixed or frozen. The fixed samples were prepared for a study of immunocytochemical staining of NGF and its two receptors. Total RNA was extracted from the frozen ovarian samples, and the expression of NGF, TrkA and p75 was investigated by RT-PCR. Products were resolved by 1% agarose gel electrophoresis and image analysis. Immunocytochemical staining revealed the expression of NGF in granulosa cells (GC) and oocytes; TrkA was mainly in oocytes and in GC in minority of the samples; and p75 was in some of the stroma cells from fetuses aged less than 22 GW. Transcripts of NGF and TrkA were identified by RT-PCR in all samples, while those for p75 were detected only in ovarian samples from fetuses aged less than 22 GW. To elucidate if NGF is indeed involved in growth initiation of human primordial follicles, it should be added to their culture medium. The immunocytochemical detection of p75 in some of the stroma cells and transcripts in ovarian samples of fetuses less than 22 GW may suggest its role in follicular assembly.

Maternal and fetal outcome of lupus pregnancy: a prospective study of 29 pregnancies.

The aim of this study was to analyse pregestational and pregnancy risk factors for adverse fetal and maternal outcome in lupus pregnancy. Twenty women with systemic lupus erythematosus (SLE) (29 pregnancies) were prospectively evaluated. Mean patient age was 29.5+/-4.7 years, and mean disease duration, 6.3+/-6.5 years. Twenty-two pregnancies (75.9%) ended in live births; preterm delivery occurred in 17.4%, intrauterine growth restriction in 50%, preeclampsia in 3.7%, and gestational hypertension in 8%. Six pregnancies (20.7%) ended in spontaneous abortions. Adverse live-birth outcome was significantly associated with low pregestational serum albumin level, elevated gestational anti-dsDNA antibody, and diabetes mellitus. Spontaneous abortion was directly associated with low levels of pregestational serum albumin, positive anticardiolipin IgA, anti-beta2-glycoprotein I IgM, and anti-La antibodies, and inversely associated with number of patients' children. Postgestational lupus flare-up was noted in six pregnancies. Risk factors included high pregestational SLE Disease Activity Index (SLEDAI), lower serum albumin, elevated serum antibody to dsDNA, proteinuria, and use of prednisone and hydroxychloroquine. We conclude that despite high rate of obstetrical complications and postpartum lupus flare-up, pregnancy poses low risk for the majority of women with SLE.

Immunocytochemical detection and RT-PCR expression of leukaemia inhibitory factor and its receptor in human fetal and adult ovaries.

The ability to mature human primordial follicles in vitro would assist fertility restoration. However, the signals initiating growth of primordial follicles are unknown. Growth factors such as leukaemia inhibitory factor (LIF) may play a role in this process. To investigate the expression of LIF and its receptor in early developing follicles, nine ovarian samples from adolescents/adults aged 13-43 years and 23 ovaries from human fetuses aged 19-33 gestational weeks were immediately fixed or frozen. The fixed samples were prepared for a study of immunocytochemical staining of LIF and its two receptor units (LIF-R and gp 130). mRNA was extracted from the frozen ovarian samples, and the expression of LIF, LIF-R and gp 130 was investigated by RT-PCR. Products were resolved by 10% polyacrylamide gel electrophoresis and image analysis. There was strong to moderate immunocytochemical staining for LIF and LIF-R in oocytes from the primordial follicular stages onwards, and very weak to moderate staining for gp 130. LIF-R was also detected in granulosa cells of primary and secondary follicles from adolescents/adults. Transcripts of LIF, LIF-R and gp 130 RNA were identified by RT-PCR in all samples. The immunocytochemical staining and mRNA expression of LIF and its receptor are consistent with the concept that LIF might be involved in growth initiation of human primordial follicles through its receptor.

Inclusion of standard and low-dose gonadotropin releasing hormone-analog (short protocol) in controlled ovarian hyperstimulation regimens in normogonadotropic patients aged 40-48 years who are undergoing in vitro fertilization.

We aimed to compare the efficiency of three controlled ovarian hyperstimulation protocols in achieving superovulation in normogonadotropic patients aged 40 years or more, who were undergoing in vitro fertilization (IVF) treatment. This was a prospective randomized clinical study, carried out in the Infertility and IVF Unit of an academic tertiary hospital. A total of 219 normogonadotropic patients (serum follicular stimulating hormone level < 15 mIU/ml) aged 40-48 years, with regular menstrual cycles, were randomly allocated to one of three short follicular protocols: menotropins only (group A), menotropins plus a mini-dose of gonadotropin releasing hormone (GnRH)-analog (600 microg/ day) (group B), or menotropins plus a standard dose (900 microg/day) of a GnRH-analog (group C). Those cycles that reached the stage of oocyte retrieval (67, 70 and 71 cycles, respectively) were analyzed. The mean daily dose of menotropins needed for ovarian stimulation was higher when GnRH-analog was used (groups B and C) (p < 0.02; ANOVA), although there was no significant difference in the time of human chorionic gonadotropin injection (average: cycle day 11). Peak estradiol levels (p < 0.02), number of oocytes retrieved (3.9, 5.4 and 5.5 oocytes/cycle, respectively, p < 0.02) and number of embryos transferred (1.6, 1.8 and 2.1 embryos/cycle, respectively, p < 0.05) were higher when GnRH-analog was included in the controlled ovarian hyperstimulation protocol. The IVF treatment resulted in 19 pregnancies (9.1% implantation rate), with a similar distribution among all three groups (11.9%, 8.6% and 7.0%). However, a higher miscarriage rate was noted in the menotropins-only group (67.5% vs. 33.3% and 40.0% of pregnancies). No differences were observed in any of the aforementioned variables between the mini-dose and standard dose GnRH-analog groups (groups B and C). In conclusion, controlled ovarian hyperstimulation before IVF treatment in normogonadotropic patients aged 40 years or more is more effective when a GnRH-analog (short protocol) is included in the treatment regimen. In this selected group of patients, reducing the daily dose of GnRH-analog does not improve the treatment results.

Adolescent varicocele: is it a unilateral disease?

OBJECTIVES: To evaluate the incidence of left and right varicocele in adolescents. METHODS: The study group consisted of 28 adolescents who underwent evaluation for varicocele at our clinic. In 19 patients, varicocele was detected on routine physical examination, and 9 patients presented with scrotal pain or discomfort. All patients were evaluated by three methods. Physical examination was followed in all cases by contact thermography and venography studies of both testes. RESULTS: The rate of left and right retrograde flow in the spermatic veins by the three methods used was as follows: physical examination 92.8% and 10.7%; contact thermography 100% and 89%; and venography 100% and 85.7%, respectively. Varicocele was not detected by physical examination in 2 (7.2%) of the 28 patients on the left side and in 21 (87.5%) of 24 on the right side. CONCLUSIONS: The main finding of this study was that varicocele is a bilateral disease in 85.7% of adolescents. The high percentage of bilateral varicocele in our sample may explain the pathophysiologic mechanism whereby what has traditionally been considered a unilateral disease can produce bilateral testicular dysfunction. The high incidence of subclinical bilateral varicocele may indicate that we should consider varicocele a bilateral disease. The second finding was that only 10% of patients with right varicocele were diagnosed by physical examination and more than 85% were diagnosed by thermography, with confirmation by venography. Therefore, we suggest that thermography and venography should play a major role in the diagnosis of varicocele.

Routine hemoglobin testing following an elective Cesarean section: is it necessary?

OBJECTIVE: To determine the clinical relevance of routine hemoglobin testing following an elective Cesarean section. METHODS: The charts of all women who underwent elective Cesarean section at a public teaching hospital during 2001 were reviewed for pre- and postoperative hemoglobin values, demographic data, indication for surgery and need for blood transfusion. RESULTS: The study group included 383 patients. Mean (+/- SD) hemoglobin level was 12.24 +/- 1.09 g/dl prior to surgery and 10.87 +/- 1.2 g/dl after, a mean loss of 1.37 +/- 0.87 g/dl (p < 0.001). There was no significant difference in blood loss by indication for surgery. None of the patients needed an intraoperative or postoperative blood transfusion. CONCLUSION: Routine postoperative hemoglobin measurement after an uncomplicated elective Cesarean section in asymptomatic low-risk women is not necessary and should be eliminated.

Subsidised oocyte donation in Israel (1998-2000): results, costs and lessons.

BACKGROUND: Israeli law stipulates that all women aged 45-51 who need oocyte donation are entitled to as many donations as necessary, up to the birth of one child. Only oocytes donated by women who themselves are undergoing assisted reproduction are allowed. The government subsidizes all oocyte donation cycles through the medical insurer of the recipient, whether or not the procedure is performed in a public or private institution. The aim of the present study was to investigate the success of oocyte donation cycles in Israel for the period 1998-2000 and to estimate costs and pregnancy rates. METHODS: Data were derived from the Dan District Registry of the General Health Services (Sheirutei Bri'ut Clalit), the largest medical insurer in Israel. RESULTS: 171 women underwent 254 oocyte donation cycles within these 3 years, and 45 babies were born, for a positive outcome of 17.7%. Average annual cycle cost was US$1742 and average annual cost per patient, US$2521. The total annual cost for the district accounted for only 0.05% of the budget. CONCLUSIONS: These findings suggest that IVF with donated oocytes is relatively efficient and that government funding of oocyte donation cycles ensures a reasonable cost.

The 'tulip sign': a sonographic clue for in-utero diagnosis of severe hypospadias.

OBJECTIVE: To describe a unique sonographic sign for prenatal detection of severe penoscrotal hypospadias. METHODS: Ultrasound findings of hypospadias diagnosed in the second trimester of pregnancy in seven patients were compared with postnatal clinical features. All patients were recruited from routine sonographic examinations performed for various obstetric indications. RESULTS: In six of the seven cases with hypospadias, a severe form of peno-scrotal hypospadias was found. In all six cases, a unique ultrasound feature was observed represented by extreme ventral angulation of the penis, with or without chordee, in a form resembling a tulip flower. This 'tulip' is formed by the ventrally bent penis located between the two scrotal folds. In all six patients, the postnatal pictures of the newborns' genitalia corresponded perfectly to the prenatal sonograms. Associated anomalies occurred in two cases, with Nager syndrome in one case and mild renal hydronephrosis in the other case. CONCLUSIONS: The 'tulip sign' is a specific ultrasonic finding of severe hypospadias. The recognition of this may help to distinguish between severe hypospadias and other genital abnormalities (e.g. ambiguous genitalia).

Follicle-stimulating hormone treatment for men with idiopathic oligoteratoasthenozoospermia before in vitro fertilization: the impact on sperm microstructure and fertilization potential.

OBJECTIVE: To assess the effect of FSH on sperm fertilization potential and sperm intracellular structure in men with oligoteratoasthenozoospermia and a proven low fertilization rate in IVF. DESIGN: Prospective, randomized, partial crossover study. SETTING: IVF Unit, Golda Campus, Rabin Medical Center, Petah Tikva, Israel. PATIENT(S): Forty normogonadotropic, normogonadal men with oligoteratoasthenozoospermia and at least one previous IVF attempt in which fertilization failed or the fertilization rate was <30%. INTERVENTION(S): The men were randomly assigned to treatment with daily injections of 75 IU of FSH or 150 IU of FSH for at least 60 days before IVF treatment. A control group of men underwent an IVF cycle without treatment and then were randomly assigned tojoin group 1A or 1B for an additional IVF cycle with treatment. MAIN OUTCOME MEASURE(S): LH, FSH, and testosterone levels during FSH treatment, evaluation of ultramorphologic changes in sperm by electron microscopy, and comparison of fertilization rates in the control and study groups. RESULT(S): After treatment with 75 IU or 150 IU of FSH, the mean fertilization rates were 19.7% and 20.5%, respectively, compared with a 5.8% fertilization rate in the study control cycles. CONCLUSION(S): Prolonged treatment with FSH results in a significant increase in fertilization rates. This effect may be related to improvements in subcellular components of the sperm.

The role of purified follicle stimulating hormone therapy in the male partner before intracytoplasmic sperm injection.

OBJECTIVE: To evaluate the impact of long-term purified FSH (pFSH) therapy in male partners before intracytoplasmic sperm injection (ICSI). DESIGN: Prospective, randomized, controlled study. SETTING: Large university-based IVF unit. PATIENT(S): Seventy-eight patients made up the study and control groups (39 patients each). All patients had severe male factor infertility. INTERVENTION(S): Induction of ovulation, oocyte retrieval, ICSI, and ET were carried out in both groups. In the study group, male partners received pFSH (75 IU FSH, <1 IU LH) for > or = 50 days before oocyte retrieval. MAIN OUTCOME MEASURE(S): Fertilization rate, embryo quality, implantation and pregnancy rates. RESULT(S): Fertilization and pregnancy rates were higher in the study group (68% and 35.9%, respectively) were higher than in the control group (59% and 17.9%, respectively), although the differences did not reach statistical significance. The implantation rate was significantly higher in the study group (15.5% versus 6.5%). The study group showed a trend toward a higher number of better-quality embryos per transfer (mean +/- SD, 2.2 +/- 1.6 versus 1.6 +/- 1.6). CONCLUSION(S): Purified FSH therapy in male partners before ICSI improves implantation rate. Improved embryo quality may be a contributory factor.

Ovarian abscess after ovum retrieval for in-vitro fertilization.

An ovarian abscess is an uncommon surgical emergency that could be lethal. The causes of an ovarian abscess vary, and treatment thereof may unfortunately lead to an oophorectomy. In order to draw the attention of physicians to this rare entity, we present a case of ovarian abscess resulting from follicle aspiration for in-vitro fertilization. Furthermore, with correct preoperative diagnosis and prompt surgical intervention at an early stage, the affected ovary may be salvageable.

Morphology and clinical outcomes of embryos after in vitro fertilization are superior to those after intracytoplasmic sperm injection.

OBJECTIVE: To compare embryos obtained after IVF and intracytoplasmic sperm injection (ICSI) regarding morphology and the likelihood of achieving clinical pregnancy. DESIGN: Case-control study. SETTING: An IVF unit controlling 1,310 cycles in 1996. PATIENT(S): Women having a total of 477 IVF and 475 ICSI consecutive cycles. INTERVENTION(S): Ovarian stimulation, IVF-ET, or ICSI-ET for all couples. MAIN OUTCOME MEASURE(S): Number of grade-A embryos transferred, preclinical pregnancy losses, and clinical pregnancy rates in IVF and ICSI cycles. RESULT(S): In comparison with the ICSI group, the IVF group showed significantly more grade-A embryos available for transfer (mean, 2 +/- 1.6 versus 1.8 +/- 1.5), significantly fewer preclinical pregnancy losses (1.6% versus 4%), and significantly higher clinical pregnancy rates (25% versus 19.1%). CONCLUSION(S): Embryos obtained after IVF are superior to those obtained after ICSI in relation to embryo morphology and the likelihood of achieving clinical pregnancy.

The impact of woman's age and sperm parameters on fertilization rates in IVF cycles.

OBJECTIVE: To examine the predictive value of different sperm parameters and the age of a couple on fertilization rate. MATERIAL AND METHODS: One-hundred six consecutive sperm samples were analysed before insemination of oocytes in IVF cycles. Sperm count and motility were examined with the Makler Counter Chamber. Morphology was evaluated by the eosin nigrosin and giemsa stains. Membrane integrity was evaluated by the hypoosmotic swelling test (HOST). RESULTS: Using a step-wise logistic regression test the following parameters were all significantly correlated with the fertilization rates: sperm concentration after a 3-h layering (chi 2 = 15.85, P < 0.001), sperm motility index (chi 2 = 6.12, P < 0.0134), woman's age (chi 2 = 6.25, P < 0.012), and the hypoosmotic test (chi 2 = 6.63, P < 0.01). A fertilization capacity formula was established by using these four parameters to predict the fertilization capacity of a couple. Significantly lower fertilization rates were detected in elderly women with the partner sperm concentration of 7 x 10(6)/ml after 3-h layering, < 30% swollen spermatozoa in the HOST and motility index score of < 50. CONCLUSIONS: By the aforementioned formula, sperm fertilization capacity can be predicted by using simple sperm parameters and women's age. Elderly women with partners who have suboptimal sperm parameters reflected by low fertilization capacity scores may benefit from micro-manipulation procedures.

The value of hysteroscopic evaluation in patients with preclinical in-vitro fertilization abortions.

The study was conducted on 144 women who experienced preclinical abortions, i.e. a transitory rise in beta-human chorionic gonadotrophin (HCG) without any clinical or sonographic evidence of pregnancy, to identify the relationship between preclinical abortions and intrauterine pathology. Hysteroscopy was performed 1-2 weeks after the decline of beta-HCG concentrations to negative values. Intrauterine adhesions were detected in three patients (2.1%), most of these being of the mild type. Concomitant intrauterine abnormalities, mainly uterine septa, were found in 14 (9.7%) cases. We believe that preclinical abortions do not predispose intrauterine adhesions and curettage is superfluous. An incomplete uterine septum seems to be the major factor predisposing this early pregnancy wastage. Hysteroscopy following this condition is an easy and efficient means for both identifying intrauterine pathology and excluding adhesions.

The use of fibrin sealant in in vitro fertilization and embryo transfer.

OBJECTIVE: To evaluate the role of fibrin sealant for embryo transfer (ET) and the effect of patient mobilization after ET on pregnancy rates. DESIGN: A prospective, randomized, controlled study. METHODS: Two hundred eleven patients who were admitted to the IVF Unit over a period of 6 months participated in the study. Patients who had three or more embryos were randomly divided into two groups: group 1 (study group), in which ET was performed using fibrin sealant, and group 2, who served as the controls. Ovulation induction was carried out using the long GnRH-a suppression protocol. RESULTS: Comparison of the results regarding the implantation and pregnancy rates and ectopic pregnancy rate revealed a nonsignificant difference between the two groups. However, analysis of the results according to the patients' age revealed a significant increase in pregnancy (P < .05) and implantation (P < .01) rate in elderly patients (aged 39-42) using fibrin sealant for ET as compared with controls. Furthermore, we found that bed rest has no advantage over patient mobilization after ET. CONCLUSION: The use of fibrin sealant for ET is advantageous in elderly women, but has no apparent effect on the success rate or ectopic pregnancy rate in younger patients. Immediate mobilization does not jeopardize the results of IVF-ET.

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

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.