The effect of activin-a on the development of mouse preimplantation embryos in vitro

Purpose: Our purpose was to clarify the involvement of transforming growth factor- (TGF-) family in the regulation of preimplantation embryo development. Methods: The effects of activin-A and TGF- on the rates of morula and blastocyst formations as well as on the cleavage velocity of a mouse two-cell embryo in vitro were analyzed. The gene expressions of these two growth factors in various developmental stages were also studied using RT-PCR. Results: Activin-A at a concentration of 0.2 ng/ml significantly stimulated not only the rate of morula formation but also the velocity of embryo cleavage, whereas no significant effect was found with TGF-. RT-PCR revealed that activin-A subunit mRNA, but not TGF- mRNA, was detected in preimplantation mouse embryo at any developmental stage. Conclusions: Activin-A plays an important role in the regulation of preimplantation mouse embryo development in an autocrine fashion.     

Unilateral renal agenesis associated with ipsilateral blind vagina

Summary Activities of DNA polymerase and were assayed in crude extracts prepared from human placenta. Polymerase activity was high in early pregnancy but low during the 2nd trimester. Polymerase activity did not change significantly with gestational weeks. The increase in polymerase activity in early pregnancy may be closely related to mitosis at the time of placental formation; there might also be some type of accelerating factor for polymerase in early pregnancy.     

Preservation of Fertility in Women Undergoing Chemotherapy: Current Approach and Future Prospects

Purpose: Anticancer treatment causes ovarian failure. Methods: Some hormones may have a protective effect on the ovary. Cryopreservation (freezing) of oocytes has had very limited success, and therefore, currently its use before chemotherapy is not a feasible option. However, cryopreservation of embryos is possible. Another solution is oocyte donation followed by in vitro fertilization (IVF). Results: Ovarian cortical slices containing primordial follicles have been cryopreserved successfully. To restore fertility, cryopreserved–thawed tissue taken from cancer patients before therapy could be replanted after recovery. The possible risk of malignancy restoration could be eliminated by obtaining unilaminar follicles from cryopreserved–thawed tissue and growing them in vitro, followed by routine IVF. Conclusions: Although women who undergo chemotherapy face limited options for fertility preservation, intensive studies in cryopreservation and in vitro maturation of follicles harbor hope for brighter prospects in the future.     

CLINICAL ASSISTED REPRODUCTION: Comparison of Recombinant and Urinary Follicle-Stimulating Hormone Preparations in Short-Term Gonadotropin Releasing Hormone Agonist Protocol for In Vitro Fertilization-Embryo Transfer

Purpose: To compare the efficiency and efficacy of two recombinant human FSH (r-FSH) and urinary (u-FSH) preparations in patients undergoing superovulation for IVF-ET using a short-term gonadotropin releasing hormone agonist (GnRH-a) (Triptorelin) protocol.Methods: A total of 88 women undergoing IVF-ET were included in this prospective study. They were randomized to receive u-FSH (150 IU/d), follitropin- (100 IU/d), or follitropin- (100 IU/d) for 2 days, and dosages were subsequently adjusted according to the ovarian response.Results: The FSH dose required for the overall stimulation was significantly lower in patients treated with r-FSH than in those treated with u-FSH while serum FSH values were higher in the latter group. There were no statistically significant differences in ovarian response and IVF outcome between r-FSH preparations.Conclusions: Recombinant FSH preparations have a higher efficiency than urinary ones in patients undergoing IVF-ET using a short-term GnRH-a protocol. In this situation, the two recombinant follitropins have comparable effectiveness.     

The first reported case of down syndrome in the Southern California Fertility Institute's in vitro fertilization and embryo transfer program

The first case of a genetic abnormality in a 3- year in vitro fertilization and embryo transfer program was diagnosed after routine amniocentesis of a 38-year-old patient. The G-banded karyotype analysis gave 47, XY, +21, which constitutes an abnormal chromosome pattern consistent with Down syndrome.     

The biological significance of phospholipase C beta 1 gene mutation in mouse sperm in the acrosome reaction, fertilization, and embryo development

Purpose: We carried out this study to evaluate the biological significance of phospholipase C 1 gene mutation in mouse sperm in the acrosome reaction, fertilization, and embryo development.Methods: Study subjects were divided into two groups according to the sperm [intact phospholipase C (PLC) 1 and PLC 1^–/– C57BL/6J × CBA F₁ mouse sperm] used. The positive acrosome reaction rate labeled with fluorescein isothiocyanate–Pisum sativum agglutinin, the fertilization rate, and the rate of embryos developed to the stage of morula or blastocyst in the two groups were compared.Results: The mouse sperm null for the PLC 1 gene showed a lower acrosome reaction rate than control sperm (69.2 vs 50.9%, P < 0.05). And the fertilization rate and the rate of embryos developed to the stage of morula or blastocyst were also lower in the group using PLC 1^–/– mouse sperm compared to the intact group (P < 0.05; 73.5 vs 51.8% and 15.7 vs 4.3%, respectively).Conclusions: Mutation of the PLC 1 gene in the mouse sperm reduces the acrosome reaction rate, fertilization rate, and embryo development rate, which may be the etiologic factors responsible for the low reproductive rate of PLC 1^–/– mouse.     

Polymorphisms for interleukin-1 beta (IL-1 beta)-511 promoter, IL-1 beta exon 5, and IL-1 receptor antagonist: nonassociation with endometriosis

Purpose: We aimed to investigate if interleukin-1 (IL-1) and IL-1 receptor antagonist (IL-1Ra) gene polymorphism could be used as markers of susceptibility in endometriosis. Materials and Methods: Women were divided into two groups: 1) endometriosis (n = 120); 2) nonendometriosis groups (n = 103). Polymorphisms for IL-1-511 promoter, IL-1 exon 5, and IL-1Ra were detected by polymerase chain reaction. Genotypes and allelic frequencies for these polymorphisms in both groups were compared. Results: Proportions of different IL-1 and IL-1Ra polymorphisms in both groups were nonsignificantly different. Proportions of C homozygote/heterozygote/T homozygote for IL-1-511 promoter in both groups were 1) 21.6/59.1/19.1% and 2) 26.2/50.5/23.3%. Proportions of E1 homozygote/heterozygote/E2 homozygote for IL-1 exon 5 in both groups were 1) 91.6/5/3.3% and 2) 95.15/4.85/0%. Allele I/II/IV/V for IL-1Ra in both groups were 1) 92.5/5.4/ 1.6/0.4% and 2) 95.1/3.9/1/0%. Conclusions: Association of endometriosis with IL-1-511 promoter, IL-1 exon 5, and IL-1 receptor antagonist gene polymorphisms doesn't exist. These polymorphisms are not useful markers for prediction of endometriosis susceptibility.     

Reduced expression of alphavbeta3 integrin in the endometrium of unexplained infertility patients with recurrent IVF-ET failures: improvement by danazol treatment

Purpose : To determine whether there is any association between the expression of endometrial integrin v 3 and repeated IVF-ET failure and to examine the effect of danazol treatment on v 3 expression. Methods : This prospective study was performed using a semiquantitative immunohistochemical analysis on the staining intensity of v 3 in the mid-secretory endometria derived from 10 fertile women and 57 infertile patients with a history of repeated IVF-ET failures. Nine patients randomly selected from these 22 patients with unexplained infertility were then treated with oral danazol administration for 12 weeks and reexamined at the first mid-secretory phase after the danazol treatment. Result(s) : The levels of endometrial v 3 expression were lower in 22 patients with unexplained infertility than in the fertile control and 35 patients with explained infertility. The 9 patients treated with danazol showed a significant increase in the v 3 staining. Conclusion(s) : The significantly decreased expression of endometrial integrin v 3 suggested that functional, but not morphological, endometrial defect may be one of the causes for the patients with unexplained infertility. Danazol may have a therapeutic potential in improving endometrial function together with up-regulation of v 3.     

The relation between depth of trophoblastic invasion and β-HCG levels in tubal pregnancies

-HCG (human chorionic gonadotropin) values of over 2500 I.U./l are associated with higher failure rates for therapy with prostaglandin F2 alpha in tubal pregnancies. The purpose of our study was to ascertain if the 2500 I.U./l limit correlates with histopathology. We therefore compared the pre-operative -HCG-values and intraluminal and extraluminal trophoblast growth in tubal pregnancy. Purely intraluminal trophoblast was significantly more frequent in patients of group I (-HCG < 2500 I.U./1), while group II patients (-HCG > 2500 I.U./l.) almost exclusively had extraluminal growth (P=0.0045). Since the efficacy of prostaglandin F2 alpha therapy depends on intact tubal musculature the correlation of the -HCG threshold level with histopathologic findings may explain the high failure rate in patients with -HCG values above 2500 I.U./l. Correspondence to: M. Klein     

Modifying effects of epidural analgesia or general anesthesia on the stress hormone response to laparoscopy for in vitro fertilization

Modifying effects of epidural analgesia and general anesthesia on stress hormone release was studied during laparoscopy for in vitro fertilization (IVF). In 24 women follicle development was stimulated by clomiphene and gonadotropin treatment, and oocytes were collected by laparoscopy under epidural analgesia in 11 women and under fentanyl-supplemented nitrous oxide-oxygen anesthesia in 13. The plasma levels of immunoreactive -endorphin (ir -E), cortisol, and prolactin (PRL) did not change under epidural analgesia per se, but after the start of laparoscopy, increased release of all these stress hormones was observed. General anesthesia per se increased the release of PRL, whereas the release of cortisol and ir -E decreased, probably due to the effects of fentanyl and thiopentone. During the stress attributed to laparoscopy, significantly more ir -E and cortisol was released under epidural than under general anesthesia, whereas the release of PRL was more significant under general anesthesia. These results show that neither mode of anesthesia prevented the stress response to laparoscopy. In the subsequent midluteal phase, the mean plasma level of progesterone and the mean progesterone-estradiol ratio were significantly greater in the epidural than in the general anesthesia group, suggesting that the mode of anesthesia may have an effect on the luteal phase. The significance of this difference on the conception rate remained unsolved, however.     

Does Pretreatment with Progestogen or Oral Contraceptive Pills in Low Responders Followed by the GnRHa Flare Protocol Improve the Outcome of IVF-ET?

Purpose: Women undergoing in vitro fertilization with lowovarian reserve and poor response to controlled ovarianhyperstimulation (COH) present a management dilemma.In a retrospective observational study, we compared thepretreatment use of the gestogen medroxyprogesterone acetate(10 mg twice daily from day 15 of the cycle for aminimum of 2 weeks) with an oral contraceptive pill (onetablet daily from day 4 of the cycle for a minimum of 3 weeks). Methods: The criteria for inclusion in the study includedone or more of the following: abandoned cycles due topoor response, fewer than four oocytes retrieved followinga standard COH protocol, age >39 years, and elevatedbasal serum follicle-stimulating hormone (FSH).Thirty-eight women received pretreatment with gestogen, and asimilar number of women received pretreatment with thepill. The flare protocol was used in all treatment cyclescombined with an individualized dose of human menopausalgonadotropin (hMG) (4–8 ampoules/day of 75 units FSH/ampoule) depending on previous response, age, and earlyfollicular serum FSH level. Both groups were similar inmean age, duration of infertility, early follicular FSH levels,and the distribution of various aetiologies. Results: Twenty-nine cycles were abandoned before oocyteretrieval, 15 (39.5%) in the pill group and 14 (36.8%) inthe gestogen group, because of an inadequate ovarianresponse. The mean (±SD) number of ampoules (75 IUFSH/ampoule) of hMG used per cycle was similar in thepill and gestogen groups (59.7 ± 19.3 vs. 70.2 ± 29.4,respectively). There also was no difference seen in the numbersof oocytes retrieved (4.4 ± 2.3 vs. 4.2 ± 2.5), totalnumber of embryos (2.5 ± 2.4 vs. 2.2 ± 1.1), or the numberof embryos transferred (1.8 ± 1.2 vs. 2.1 ± 1.0) in the pilland gestogen groups, respectively. One pregnancy in eachgroup resulted following embryo transfer in 22 women inthe pill group and in 24 women in the gestogen group. Conclusions: We conclude that pre-IVF treatment with oralcontraceptive pill or gestogen combined with the flare protocolin women at high risk of or with a history of poor ovarianresponse, as defined in this study, did not appear to resultin an improvement in outcome of IVF-embryo transfer.     

Polymorphism in the tumor necrosis factor-alpha gene in women with preeclampsia

Purpose : We determined whether genetic variability in the gene encoding for tumor necrosis factor- (TNF-) contributes to individual differences in susceptibility to the development of preeclampsia. Methods : The study involved 133 preeclamptic and 115 healthy control pregnant women who were genotyped for C-850T polymorphism in the TNF- gene promoter. Chi-square analysis was used to assess genotype and allele frequency differences between preeclamptic women and controls. Results : A significantly different genotype distribution of C-850T polymorphism was observed between the two groups, with the frequency of the variant T allele being significantly reduced in the preeclamptic group (4.5%) when compared with the control group (9.6%) (P = 0.03; OR = 0.45, 95% CI = 0.22–0.92). Accordingly, the odds ratio for preeclampsia associated with the pooled TT and CT genotypes was 0.367 (P = 0.02; 95% CI = 0.159–0.847). Conclusions : The T allele of the TNF- gene may modify individual preeclampsia risk, being protective against the development of the complication.     

Lokal- und Leitungsanästhesien in der Geburtshilfe

Zusammenfassung Die rückenmarknahe Regionalanalgesie ist das effektivste Analgesieverfahren bei der vaginalen Entbindung. Werden hierbei Lokalanästhetika mit Opioiden kombiniert, kann in vielen Fällen eine motorische Beeinträchtigung der Mutter vermieden werden. Die Pudendusanästhesie eignet sich zur schmerzfreien Anlage und Naht einer Episiotomie und bei vaginal-operativen Entbindungen. Die Spinalanästhesie ist für die meisten elektiven Kaiserschnitte ein gut geeignetes Verfahren. Bei dringlichen Kaiserschnitten kann sie statt einer risikoreicheren Allgemeinanästhesie durchgeführt werden. Die Epiduralanästhesie erlaubt eine langsamere Ausbreitung des Anästhetikums und ist deshalb z. B. bei bestimmten mütterlichen Vorerkrankungen gut geeignet. Voraussetzung für die sichere Anwendung der rückenmarknahen Analgesie- und Anästhesieverfahren sind die Prophylaxe und schnelle Therapie von Nebenwirkungen und Komplikationen sowie die Kenntnis von Kontraindikationen.     

Epidermal growth factor (EGF) receptor localization in cultured human granulosa lutein cells and the stimulation of progesterone production by EGF and transforming growth factor-α (TGF-α)

Purpose The purpose of this study was to investigate the expression of EGF receptor (EGF-R) in human granulosa cells undergoing luteinization and progesterone production by these cells in response to EGF and TGF- alone or in combination with luteinizing hormone (LH). Granulosa cells were obtained from IVF patients following oocyte retrieval 34 to 36 hr post-hCG injection. EGF receptor was localized in cells by means of immunoperoxidase staining using a polyclonal primary antibody directed against the human EGF-R. To assess progesterone production, cells were seeded overnight, washed, and cultured with the growth factors ± LH. Medium and treatments were changed every 24 hr for 3 days.Results Specific EGF-R staining was observed in the cultured cells compared to those incubated with antibody that was preabsorbed with a 10-fold excess of EGF. Basal progesterone accumulation per 24-hr period was stimulated dose dependently on each day of culture, by both EGF (up to 3.5-fold at 5 or 50 ng/ml) and TGF- (up to 4-fold at 50 ng/ml). The addition of LH alone also stimulated progesterone accumulation daily, and this effect was further enhanced dose dependently by cotreatment with EGF or TGF-. Furthermore, tyrphostin, an EGFR-specific tyrosine kinase inhibitor, inhibited both basal and growth factor-stimulated progesterone production.Conclusion These data suggest an EGF receptormediated physiological role for EGF and TGF- in human luteal function involving an autocrine and/or a paracrine stimulation of progesterone production.Presented at the 50th Annual Meeting of the American Fertility Society, November 1994, San Antonio, Texas.     

Rapid detection of the deltaF508 mutation in single cells using DHPLC: implications for preimplantation genetic diagnosis

Purpose: Practice of preimplantation genetic diagnosis (PGD) requires efficient amplification and analysis techniques. We have tested Denaturing High Performance Liquid Chromatography (DHPLC) to screen the F508 mutation in heterozygous single cells in order to assess its usefulness for PGD of cystic fibrosis. Methods: One hundred and two single lymphocytes—including N/N and N/F508—were studied. F508 locus was amplified by nested PCR followed by the analysis of PCR products by DHPLC in non-denaturing conditions. Results: On the basis of PCR-amplified product analysis, total efficiency of amplification was 98.78% (101/102), and allele dropout (ADO) rate was 3.7% (3/81). For each sample, results were obtained in less than 4 min with high resolution. Conclusions: DHPLC is a rapid and efficient technique to detect the F508 mutation in single cells and is therefore appropriate for clinical application of preimplantation genetic diagnosis of cystic fibrosis.     

Platelet specific protein (β-thromboglobulin and platelet factor 4) in normal pregnancy and in pregnancy complicated by preeclampsia

Summary Pregnancy is accompanied by significant alterations of platelet function. Platelet activity can be determined by measurement of plasma levels of secreted platelet proteins. In this study we determined plasma levels of -Thromboglobulin (-TG) and Platelet Factor 4 (PF4) simultaneously in 35 women with uncomplicated pregnancy and in 15 patients with preeclampsia in third trimester of gestation. Additionally, PF4 plasma levels were measured using a commercially available Radio Immunoassay (RIA) and an Enzyme Immunoassay (EIA) simultaneously and values obtained were compared. Platelet count and creatinine were in the normal range in both groups; however, significantly higher levels of -TG (P<0.0005) and PF4 (P<0.0001) were found in case of preeclampsia. High levels of platelet proteins emphazise the active role of the platelets in the alterations of hemostasis in cases of preeclampsia.     

Human early pregnancy factor: serum concentrations before and after therapeutic abortion in comparison with β-hCG and an early pregnancy associated protein

Summary Serum concentrations of the early pregnancy factor (EPF), -hCG and the early pregnancy associated protein (EPAP) were measured in 12 patients before and after therapeutic abortion for social-medical reasons. Detection of EPF was performed by the rosette-inhibition assay, -hCG quantification by radioimmunoassay and EPAP measurements by rocket immunoelectrophoresis using a monospecific polyclonal rabbit antiserum. The disappearance rate of EPF after termination of pregnancy was closely correlated with the decrease of -hCG concentrations and loosely correlated with the decrease of EPAP concentrations . No correlation has been found between hCG and EPAP values.     

Adenomyosis in a patient with mosaic Turner’s syndrome

Background Few case reports describing endometriosis in patients with gonadal dysgenesis have been published, but none has reported the presence of adenomyosis in a patient with Turners syndrome.Case A 31-year-old woman with mosaic Turners Syndrome (45,X/46,XX/47,XXX) was referred to us because of severe iron deficiency anaemia due to hypermenorrhea and persistent lower abdominal pain for more than six months. She presented normal secondary sex development, normal breast, normal pubic and axillary hair. The external genitalia were also normal. Laboratory examination showed normal gonadotropin, 17beta-estradiol, plasma androgens and cortisol levels. At transabdominal ultrasound a myoma (15×8.5×8 cm) arising from the posterior uterine wall was suspected. The mass was removed during laparotomy. Histologic examination confirmed the presence of the myoma and revealed the presence of focal adenomyosis.Conclusion Adenomyosis and leiomyomata are separate entities but they share a common pathology in that they develop primarily in women of reproductive age and their growth is oestrogen dependent. To our knowledge, this is the first case report in the literature of adenomyosis in a woman who had the Turners syndrome.     

A case of Ashermann’s syndrome following surgical evacuation of uterus for secondary postpartum haemorrhage

Surgical evacuation of uterus for retained products of conception following delivery is a very common procedure but can have important sequelae. The possibility of causing Ashermanns syndrome, although rare, should always be borne in mind. If severe enough, it can render a woman essentially infertile, which can have serious psychological sequelae. Hysteroscopic recavitation is not only complex surgery, but it is also associated with potential problems even when successful, so realistic counselling is essential.