[Recommendations for estrogen and progesterone replacement in climacteric and postmenopause. 33. Workshop of the "Zurich Discussion Circle", October 2004].
Explore the source record for details and available documents.
Aged↗
Biomedical subjects
J Neulen
Publications and source records attributed to J Neulen.
[Recommendations for hormonal contraception. 34. Workshop of the "Zurich Discussion Circle", April 2005].
Explore the source record for details and available documents.
Aged↗
Regulation of gene expression in endothelial cells: the role of human follicular fluid.
A precise regulation of angiogenesis is a prerequisite for an adequate maturation of ovarian follicles. Despite the production of vascular endothelial growth factor (VEGF) by granulosa cells in antral follicles, angiogenesis is restricted to the theca cell layer. The maturing follicle remains avascular before ovulation, implying regulatory mechanisms which prevent premature follicular vascularization. In order to investigate the role of follicular fluid and of granulosa cells in the regulation of endothelial gene expression, human umbilical vein endothelial cells (HUVECs) were incubated in vitro with media conditioned with human follicular fluid obtained from individual patients undergoing oocyte retrieval for in vitro fertilization procedures or with culture medium conditioned by human granulosa cells respectively. Using microarray technology, the gene expression pattern was compared between untreated monolayers of HUVECs and HUVECs treated either with follicular fluid or with granulosa cell conditioned media. We identified a total of 15 genes that were significantly up-regulated and 11 genes that were significantly down-regulated in endothelial cells treated with follicular fluid at least 2.5-fold in more than 70% of comparisons. Up-regulated genes involved in angiogenesis were the anti-angiogenic factors gro-beta (16.5-fold), angiopoietin-2 (3.9-fold), alpha-2-macroglobulin (24.3-fold) and the pro-angiogenic factors E-selectin (5.3-fold) and vascular cell adhesion molecule-1 (VCAM-1) (4.4-fold), whereas a significant down-regulation of the pro-angiogenic genes fibulin-5 (3.5-fold) and elastin (14.9-fold) could be observed. Culturing of HUVECs with conditioned medium from cultured human luteinized granulosa cells demonstrated a similar regulatory pattern of gene expression for fibulin-5, elastin, gro-beta, and E-selectin. The gene regulation in endothelial cells by follicular fluid could be confirmed by RT-PCR for gro-beta, angiopoietin-2, elastin, fibulin-5, and E-selectin. The present work reveals that compounds secreted by granulosa cells lead to the expression of anti-angiogenic factors on the transcript level in endothelial cells and thus could help to explain the temporal and spatial discrepancy between the high expression of VEGF and the restricted angiogenesis in the preovulatory follicle.
Down-Regulation↗
[Fluoroscopically guided fallopian tube recanalization with a simplified set of instruments].
PURPOSE: Fluoroscopically guided transcervical fallopian tube recanalization is recognized as an important step in the workup of female infertility. In the present study, a simplified set of recanalization instruments was tested. MATERIALS AND METHODS: Forty-two women with infertility and sonographically confirmed or suspected uni- or bilateral tubal occlusion were examined. After vaginal placement of a plastic speculum and fixation of a tenaculum, a 4F glide catheter with a 0.89 mm glidewire was advanced transcervically. After documentation of tubal occlusion by hysterosalpingography, the uterotubal junction was catheterized with the same instruments. Under fluoroscopic guidance, the glidewire was negotiated beyond the intramural portion of the tube. Selective salpingography documented the outcome of the recanalization. RESULTS: Hysterosalpingography confirmed tubal occlusions in 26 of 42 patients (in 12 cases unilateral and in 14 cases bilateral). Fallopian tube recanalization was successful in 23 of 26 patients (technical success rate of 88 %). The resulting fertility rate was 30 % (7/23), without any ectopic pregnancy. Complications such as tubal perforation, infection, or bleeding did not occur. Tubal catheterization was straightforward and smooth in 17/23 cases. For a hyperflexed uterus (6/23), a curved tip of the catheter was helpful in tubal probing. CONCLUSION: Fluoroscopically guided transcervical fallopian tube recanalization is a patient-friendly alternative to tubal microsurgery and in-vitro fertilization. Tubal catheterization with 4F glide catheter and glidewire further streamlines the procedure.
Adult↗
[Recommendations for estrogen and progestin replacement therapy in climacteric and postmenopause. 31. Professional Meeting of the Zurich Discussion Circle, October 2003].
Explore the source record for details and available documents.
Adult↗
Effects of blood estrogen level on cortical activation patterns during cognitive activation as measured by functional MRI.
Modulation of the blood estrogen level as it occurs during the menstrual cycle has a strong influence on both neuropsychological and neurophysiological parameters. One of currently preferred hypotheses is that the menstrual cycle hormones modulate functional hemispheric lateralization. We examined six male and six female subjects by functional magnetic resonance imaging (fMRI) to image cortical activation patterns associated with cognitive and motor activation to determine whether these changes during the menstrual cycle can be visualized. Female subjects, who did not use oral contraceptives, were scanned twice, once during the menses and once on the 11/12 day of the menstrual cycle. A word-stem-completion task, a mental rotation task and a simple motor task were performed by all subjects. Our data provide evidence that the menstrual cycle hormones influence the overall level of cerebral hemodynamics to a much stronger degree than they influence the activation pattern itself. No differences were seen between male subjects and female subjects during the low estrogen phase. During both neuropsychological tasks blood estrogen level had a profound effect on the size but not on the lateralization or the localization of cortical activation patterns. The female brain under estrogen showed a marked increase in perfusion in cortical areas involved in both cognitive tasks, whereas the hemodynamic effects during the motor tasks were less pronounced. This might be due to differences in neuronal or endothelian receptor concentration, differences in synaptic function, or, most likely, changes in the cerebrovascular anatomy in different cortical regions.
Adult↗
Poor responder-high responder: the importance of soluble vascular endothelial growth factor receptor 1 in ovarian stimulation protocols.
This study was designed to detect vascular endothelial growth factor (VEGF) and its soluble receptor (sVEGFR-1) in follicular fluid specimens and to evaluate the importance of sVEGFR-1 with respect to ovarian response to gonadotrophin stimulation. A total of 69 patients was treated for IVF with recombinant human follicle stimulating hormone (FSH). Concentrations of VEGF and sVEGFR-1 were quantified in follicular fluids from oocyte retrievals. Patients were designated to three groups with respect to the number of harvested oocytes: group A, 1-5 oocytes; group B, 6-10 oocytes; group C, >10 oocytes. In group A, 1133 +/- 870 pg VEGF/ml follicular fluid per oocyte were quantified, in group B 426 +/- 262 pg VEGF/ml per oocyte, and in group C 274 +/- 179 pg VEGF/ml per oocyte. Soluble VEGFR-1 concentrations resulted in 1200 +/- 523 pg/ml follicular fluid per oocyte in group A, 255 +/- 193 pg/ml per oocyte in group B, and 79 +/- 69 pg/ml per oocyte in group C. No free sVEGFR-1 could be detected in any follicular fluid. An index to estimate the biological activity of VEGF by dividing VEGF/sVEGFR-1 revealed an increasing availability of VEGF with higher ovarian response to gonadotrophin therapy. In group A this index was 1.03, in group B 1.71, and in group C 3.21. A delicate balance between VEGF and sVEGFR-1 is necessary to allow an adequate ovarian reaction to gonadotrophin therapy. Excess of bio-active VEGF increases the risk for ovarian hyperstimulation syndrome. Excess of sVEGFR-1 results in poor response and goes in parallel with reduced chances for conception.
Adult↗
[Gonadotropin stimulation of progesterone production of granulosa cells derived from in vitro fertilization procedures: comparison of the in vitro biopotency of recombinant follicle stimulating hormone, recombinant luteinizing hormone nad recombinant human chorionic gonadotropin].
The aim of this study was to investigate the effects of recombinant human gonadotrophins (follicle stimulating hormone [rFSH], luteinizing hormone [rLH] and human chorionic gonadotrophin [rhCG]) on granulosa-lutein cell progesterone production in long-term culture. Cultures were maintained for 9 days including a preincubation period for 3 days. Cells obtained from gonadotrophin-stimulated cycles produced increasing amounts of progesterone during the preincubation period reaching a maximum concentration on day 3. Granulosa cells in absence of serum secreted significantly lower levels of progesterone than in its presence and showed moderate responses to rhCG. Addition of serum (foetal calf serum, FCS) to the culture medium enhanced progesterone output in both control and rhCG-stimulated cultures in a dose-dependent manner. However, in absence of rhCG, granulosa cell progesterone production declined towards the end of culture even in the presence of constantly high FCS levels. All three recombinant gonadotrophins stimulated progesterone accumulation. Recombinant FSH and rLH, applied in the dose interval of 0.001-0.01-0.1 IU/ml, caused clear dose-related increases in progesterone production. Progesterone accumulation was also significantly augmented by the presence of rhCG (range of doses 0.01-0.1-1-10 IU/ml), but this effect was dose-dependent only between 0.1-10 IU/ml dose intervals with the maximum stimulation occurred at the dose of 0.1 IU/ml rhCG. From our results it can be concluded that granulosa cells require both serum supplementation and gonadotrophin stimulation for optimal progesterone synthesis. Recombinant FSH, completely devoid of LH activity, was equivalent to rLH and rhCG in terms of stimulation of progesterone production of luteinized human granulosa cells.
Cells, Cultured↗
Elevated serum progesterone concentrations during the early follicular phase of the menstrual cycle: clinical significance and therapeutic implications.
This study describes the incidence and clinical significance of hyperprogesteronemia in the early follicular phase of the menstrual cycle and the effects of prednisolone treatment in patients with this condition. Progesterone, testosterone and dehydroepiandrosterone sulfate (DHEAS) serum concentrations were determined in 316 consecutive patients admitted for infertility treatment. Elevated concentrations of progesterone in the early follicular phase could be detected in 36 cases (11.4%). In these patients, elevated testosterone and DHEAS concentrations could be detected in eight and ten cases, respectively. Serum concentrations of 17 alpha-hydroxyprogesterone and cortisol were normal in all patients. Treatment with 2.5-7.5 mg prednisolone/day normalized progesterone serum concentrations within 4-8 weeks. Twenty-four patients achieved a pregnancy either by timed intercourse or by intrauterine insemination (IUI) over the following 12 months.
Adult↗
[A fluoroscopy-controlled transcervical fallopian tube recanalization].
Explore the source record for details and available documents.
Adult↗
Aneuploidy in human granulosa lutein cells obtained from gonadotrophin-stimulated follicles and its relation to intrafollicular hormone concentrations.
Proliferation of granulosa cells is inversely related to differentiation and hormone production. The purpose of this study was to evaluate the intrafollicular and serum steroid concentrations and to compare these results to granulosa cell proliferation as measured by DNA flow cytometry. Human granulosa lutein cells in follicular fluid of in-vitro fertilization (IVF) patients were investigated with regard to ploidy, percentage of S-phase cells and proliferation index (PI: percentage of cells in the S- and G2/M-phase). The study was originally designed to indicate an additional marker for the outcome of IVF treatment by DNA flow cytometric measurements of granulosa lutein cells. Follicular fluids of 160 follicles (45 patients) were evaluated; 45.6% (n = 73) of the follicles showed aneuploid granulosa lutein cells and 5.6% (n = 9) of the follicles contained multiploid granulosa cells, defined as at least two aneuploid populations of cells with different DNA indices. A total of 48.8% (n = 78) of the follicles had only diploid cells. Thus >50 % of the investigated follicles showed aneuploidy. In all, 73% (33 of 45) of patients had at least one follicle containing aneuploid granulosa lutein cells. The PI of the aneuploid cell populations significantly exceeded that of the diploid cell populations (median: aneuploid: 15.5; diploid: 7.4; P < 0.0001). The intrafollicular concentrations of testosterone, progesterone and dehydroepiandrosterone sulphate (DHEA-S) were significantly lower in follicles with aneuploid granulosa cell populations. Luteinizing hormone concentration was significantly higher in follicles with aneuploid granulosa cells. Intrafollicular concentrations of oestradiol, follicle stimulating hormone and the serum concentrations of all steroid hormones did not show any significant correlation to ploidy. Although aneuploidy has been reported for oocytes (in approximately 17% of the oocytes), no study, to our knowledge, has observed such a high incidence of aneuploidy in granulosa lutein cells after gonadotrophin stimulation. Except for aneuploidy found in tissues with some characteristics of neoplastic growth (colon adenoma, borderline tumours, endometriosis with atypic cells, etc.), it is unique for non-malignant human cells. The correlation with intrafollicular steroid concentrations points to a possible pathophysiological or physiological relevance of these findings. However, it was impossible to correlate the outcome of IVF with DNA flow cytometry results.
Adult↗
Oestradiol production by luteinized human granulosa cells: evidence of the stimulatory action of recombinant human follicle stimulating hormone.
In this study the effect of recombinant human follicle stimulating hormone (rFSH) on oestradiol production by human granulosa-lutein cells was examined in long-term culture, in the presence or absence of androgens. Cells were harvested at the time of follicular aspiration after ovarian hyperstimulation for in-vitro fertilization and cultured for 9 days. Granulosa cells were capable of secreting oestradiol spontaneously even without androgen and gonadotrophin support. Basal oestradiol secretion was relatively high and variable (421.3 +/- 159.3 pg/ml, mean +/- SEM, n = 13) on the first day and decreased gradually to 16.7 +/- 3.1 pg/ml on day 9. Addition of androgens (testosterone or androstenedione) to the incubation medium enhanced dose-dependently basal oestrogen production on days 5, 7 and 9. The androgen/oestrogen conversion rate remained constantly high during the culture, even without rFSH. After pre-incubation for 3 days, addition of rFSH resulted in a dose- and time-dependent increase in granulosa-lutein cell oestrogen production in the absence of exogenous androgens. Testosterone supplementation caused considerably higher basal oestradiol concentrations, however rFSH failed to further stimulate the oestrogen release from granulosa-lutein cells, suggesting that these cells cultured in vitro possess high aromatase activity even without rFSH support.
Androstenedione↗
Secretion of vascular endothelial growth factor/vascular permeability factor from human luteinized granulosa cells is human chorionic gonadotrophin dependent.
Vascularization is a prominent event during corpus luteum formation, providing low density lipoproteins for steroid biosynthesis and enabling transport of secreted steroids. The process of vascularization is controlled by specific regulators. Vascular endothelial growth factor (VEGF), otherwise named vascular permeability factor (VPF), induces endothelial cell proliferation as well as angiogenesis in vivo and increases capillary permeability. Here we report the expression of VEGF/VPF mRNA by cultured human luteinized granulosa cells (GC) for at least 10 days. Without HCG VEGF/VPF expression declined after day 4 and by day 10 was reduced to approximately 30% of the value at day 4. However, after culture in the presence of 1 U/ml human chorionic gonadotrophin (HCG), expression of VEGF/VPF mRNA by GC was four times greater than control experiments by day 10, and increased 100% from day 4 to day 10. Simultaneously, HCG supplementation increased VEGF/VPF secretion by GC. Medium VEGF/VPF on day 3 was 13 pM without and 11 pM with HCG. Medium VEGF/VPF on day 10 was 6 pM without HCG and 29 pM with HCG. These results suggest that vascularization of the corpus luteum is induced by HCG-mediated effects of VEGF/VPF.
Adult↗
Vascular endothelial growth factor (VEGF)/vascular permeability factor (VPF) production by luteinized human granulosa cells in vitro; a paracrine signal in corpus luteum formation.
Vascularization is a prerequisite for corpus luteum formation. Angiogenesis is thought to be regulated by vascular growth factors. Vascular endothelial growth factor (VEGF)/vascular permeability factor (VPF) specifically induces endothelial cell proliferation as well as angiogenesis and increases capillary permeability. Recently, VEGF/VPF-mRNA expression was demonstrated in luteinized human granulosa cells (GC) in vitro. In addition, the production of VEGF/VPF by human granulosa can be demonstrated immunocytochemically. VEGF/VPF is thought to mediate its effects through specific cell surface receptors. So far, two VEGF/VPF-receptors (VEGF/VPF-R) have been identified (KDR, and flt-1). A third receptor (flt-4) is highly correlated to KDR and flt-1, but the true ligand for this receptor is still unknown. The appearance of all three receptors is more or less restricted to endothelial cells. To clarify whether VEGF/VPF acts in an auto- or paracrine fashion in human luteinized GC, mRNA was scrutinized for specific expression of the three receptors by Northern blot technique. No specific VEGF/VPF-R or flt-4 transcripts were detectable, indicating that VEGF/VPF is a genuine paracrine growth factor from human luteinized GC directed to endothelial cells.
Adult↗
Non-competitive anti-oestrogenic actions of progesterone antagonists in primate endometrium: enhancement of oestrogen and progesterone receptors with blockade of post-receptor proliferative mechanisms.
Previous studies have shown that the progesterone antagonists (antiprogestins) inhibit oestrogen-dependent endometrial proliferation in ovariectomized monkeys, without having affinity to the oestrogen receptor (ER). This study was designed to investigate the effect of the antiprogestins mifepristone (RU 486) and onapristone (ZK 98,299), on the concentration of ER and progesterone receptor (PR) in the endometrium of long-term ovariectomized cynomolgus monkeys (Macaca fascicularis). In untreated monkeys, tissue preparations bound in total 228 +/- 68 pmol [3H]-oestradiol/g protein (ER), and 119 +/- 42 pmol [3H]-R5020/g protein (PR). These values were not significantly different from the total binding capacities of tissues from monkeys treated with RU 486 alone or primates treated with oestradiol plus progesterone. Treatment with oestradiol alone almost doubled the ER and PR concentrations. Combined treatment with oestradiol and RU 486 enhanced the ER and PR concentrations in a dose-dependent manner: 1 mg/kg body weight (bw) RU 486/kg increased both ER and PR contents about 3-fold. The dose of 5mg/kg bw RU 486 or onapristone increased the ER and PR concentrations almost 6- and 5-fold respectively, compared with the oestradiol-treated controls. Our results demonstrate that RU 486 and onapristone increased the endometrial ER and PR concentrations far beyond the physiological level in ovariectomized, oestradiol-treated monkeys. Whether the over-expression of ER and PR in the presence of antiprogestins is causally related to the antiproliferative impact of antiprogestins in the endometrium (non-competitive anti-oestrogenic effects) or is an independent action in unknown.
Animals↗
Ovarian stimulation in women undergoing in-vitro fertilization and embryo transfer using recombinant human follicle stimulating hormone (Gonal-F) in non-down-regulated cycles.
In order to assess the efficacy and safety of recombinant human follicle stimulating hormone (FSH) in routine clinical use, ovarian stimulation with recombinant human FSH was performed in 71 patients prior to in-vitro fertilization (IVF) without gonadotrophin-releasing hormone (GnRH) analogues in a multicentre, non-comparative study. Human chorionic gonadotrophin (HCG) was administered to 58 patients (81.7%), 15 of whom underwent 19 cycles with an initial dosage of three ampoules daily of recombinant FSH and 43 of whom underwent 152 cycles with four ampoules daily from day 3 onwards. No significant differences were detected between these two groups in all test parameters. The mean duration of treatment was 9.06 and 8.86 days respectively with a mean number of 24.06 and 23.25 vials of recombinant human FSH administered. A mean number of 6.26 and 5.88 oocytes respectively was collected. The number of transferred embryos was 2.4 and 2.2. A clinical pregnancy rate of 23.8% (10 out of 42) per transfer was achieved (30.9 and 20.6% respectively). Local tolerance of s.c. administration was excellent. Mild pain at the injection site was the dominant finding in < 20% of patients. Two cases of ovarian hyperstimulation syndrome were noted. Recombinant human FSH is very attractive to patients because it can be self-administered s.c. and the preparation does not come from a human source. In conclusion, these data support the safety and efficacy of recombinant human FSH in routine use for IVF.
Adolescent↗
Human chorionic gonadotropin-dependent expression of vascular endothelial growth factor/vascular permeability factor in human granulosa cells: importance in ovarian hyperstimulation syndrome.
Ovarian hyperstimulation syndrome (OHSS) is a severe complication arising from controlled ovarian stimulation treatment. This iatrogenic condition is potentially lethal and occurs in 0.3-5% of stimulated ovarian cycles. hCG exacerbates OHSS. The pathophysiology of OHSS is still unknown; therefore, treatment regimens are aimed at ameliorating symptoms. Prominent features of OHSS are an elevated risk of thromboembolism due to enhanced production of von Willebrand factor by endothelial cells and ascites, or pulmonary edema due to increased vascular permeability followed by third space fluid accumulation. Both of these sequelae can be evoked by vascular endothelial growth factor (VEGF), also known as vascular permeability factor (VPF). High concentrations of VEGF/VPF have been demonstrated in ascitic fluid from patients with OHSS, but the source of VEGF/VPF in these patients remained unidentified. Here we report that the messenger ribonucleic acid expression of VEGF/VPF in human luteinized granulosa cells (GCs) is dose and time dependently enhanced by hCG in vitro. Furthermore, VEGF/VPF proteins are produced by GCs. Our results suggest that the effects of hCG on the development and course of OHSS may be mediated by the production of VEGF/VPF by GCs.
Adult↗
Placental progesterone, prostaglandins and mechanisms leading to initiation of parturition in the human.
Present knowledge allows the identification of some features of the initiation of human parturition. Progesterone reduces myometrial sensitivity to labour-inducing agents. It suppresses gap junction formation and facilitates beta-adrenergic receptor expression by the myometrium which, in turn, exerts a positive feedback by enhancing beta-adrenergic-induced increases in placental progesterone production. Inhibition of gestagen action does not result in immediate initiation of labor but sensitises myometrial cells to contraction-inducing agents. Estrogens, in contrast, enable the myometrium to prepare for parturition by inducing oxytocin receptors and this seems to be the first step towards parturition. Coordinated myometrial contractions are facilitated by the increased gap junctions due to the estrogen drive. Absence of estrogen will result in failed parturition. The myometrium seems to be sensitised to oxytocin by placental CRF. Myometrial CRF receptors increase their avidity for CRF with ongoing pregnancy. Oxytocin evokes a variety of auto- and paracrine events which culminate in increased free intracellular calcium and the consequent contractions. In this cascade, prostaglandins can be identified as positive feedback agents, as they further enhance estrogen-induced expression of oxytocin receptors. Another second messenger of oxytocin action are the inositol phosphates which can further increase free intracellular calcium concentrations. Finally, endothelin-1, derived from endometrium and decidua, under oxytocin control, may serve as a myometrial contractor following delivery when oxytocin concentrations decline but when a strong myometrial contraction is needed to prevent large blood loss during and after placenta expulsion.
Female↗