Burghard Abendstein

Basic Fibroblast Growth Factor Synthesis by Human Peritoneal Mesothelial Cells: Induction by Interleukin-1

Peritoneal mesothelial cells are uniquely located to regulate cellular events in the peritoneal cavity and are an important source for various cytokines and growth factors. This study was conducted to analyze the capacity of human peritoneal mesothelial cells (HPMCs) to synthesize and release basic fibroblast growth factor (bFGF) and to characterize its regulation by inflammatory cytokines. HPMCs constitutively synthesized and released considerable amounts of bFGF as detected by a specific immunoassay. Almost 80% of bFGF (1547 +/- 173 pg/10(5) cells) was localized intracellularly. Approximately 20% of the bFGF (357 +/- 27 pg/10(5) cells) was associated with extracellular matrix components on the HPMC surface. Small amounts of bFGF (<1%) were detectable in tissue culture supernatants (8.4 +/- 1.4 pg/10(5) cells). Treatment of HPMCs with interleukin-1beta (IL-1beta; 1 ng/ml) resulted in a significant increase in bFGF production. The intracellular bFGF content showed a rapid but only transient increase, which was significant above background levels after 24 hours (41% increase; P < 0.05). This increase in intracellular bFGF concentration was associated with an induction of the release of bFGF. Within 96 hours, the release of bFGF to the cell surface and into the supernatant increased by 58% (564 +/- 52.4 pg/10(5) cells; P < 0.01) and by 214% (26.4 +/- 3.2 pg/10(5) cells; P < 0.001), respectively. Neither tumor necrosis factor-alpha nor interferon-gamma affected bFGF synthesis by HPMCs. Stimulation of HPMCs with IL-1beta increased steady-state levels of bFGF-specific mRNA. Immunohistochemical analyses of peritoneal tissue revealed constitutive expression of bFGF by HPMCs. This in situ expression proved to be most pronounced in areas of serosal inflammation in activated HPMCs. Our study demonstrates that HPMCs synthesize and release significant amounts of bFGF and that the expression of this growth factor is significantly up-regulated by the proinflammatory cytokine IL-1beta. The data support the view that HPMCs are key regulators of abdominal disease processes such as peritonitis, peritoneal fibrosis, or peritoneal tumor metastasis.

Peritoneum and Tissues of the Female Reproductive Tract as Physiological Sources of CA-125

The origin of physiological CA-125 serum levels, which in normally menstruating women were shown to depend on their actual menstrual cycle phase, has not yet been completely elucidated. It is furthermore conceivable that physiological CA-125 sources may contribute to serum elevations in the various pathologies associated with increased circulating CA-125. The present review deals with menstrual cycle-dependent expression of CA-125 in normal tissues of the female reproductive tract in relation to the actual circulating CA-125 levels together with in vivo data concerning the inductive effect of medroxyprogesterone acetate on circulating CA-125 studied in 24 postmenopausal women. Furthermore, in vitro results on constitutive, steroid hormone- and cytokine-modulated CA-125 shedding from human peritoneal mesothelial and ovarian surface epithelial cells are summarized.

Prolactin gene expression and prolactin protein in premenopausal and postmenopausal human ovaries

OBJECTIVE To investigate intraovarian prolactin and prolactin-receptor gene expression and to assess local prolactin synthesis with emphasis on possible differences between premenopausal and postmenopausal status. DESIGN The RNA extracted from human premenopausal and postmenopausal tissues was subjected to reverse transcription and polymerase chain reaction by using prolactin-specific intron- and exon-spanning primers. Prolactin-receptor expression was investigated accordingly. The amplified complementary DNA fragments were analyzed by gel electrophoresis and restriction enzyme mapping. Local prolactin hormone synthesis was verified by a time-resolved immunofluorometric assay based on our monoclonal antibodies. RESULT(S) Prolactin and prolactin-receptor gene expression was observed in all analyzed human ovaries (n = 18). Several other human tissue specimens, such as lung and kidney, served as negative control tissues. Significantly elevated concentrations of prolactin were detected in cytosolic extracts of premenopausal (n = 6; mean +/- SD; 20.6 +/- 3.3 ng/g tissue wet weight) versus postmenopausal (n = 6; 3.6 +/- 3.0 ng/g tissue wet weight) ovaries. CONCLUSION(S) The human ovary not only serves as a target for endocrine prolactin action but also as a site of local prolactin hormone production. In agreement with previous reports on extrapituitary sources of prolactin, we consider prolactin as a hormone as well as an autocrine or paracrine growth or regulatory factor. Significantly increased concentrations of prolactin in premenopausal ovarian tissue verifies its role in human reproduction.

Prevention of severe ovarian hyperstimulation syndrome (OHSS) in IVF patients by steroidal ovarian suppression — A prospective randomized study

Ovrian hyperstimulation syndrome (OHSS) is the major complication associated with artificial reproductive technologies. The aim of this study was to assess the potential of exogenous steroids to prevent OHSS without adversely affecting pregnancy rate. A prospective and randomized study including 945 consecutive cycles in 603 women (mean age 28.6 years) was carried out to evaluate the efficiency of high-dose progesterone and oestradiol administration during the luteal phase to prevent OHSS. After ovulation induction, patients were allocated by a series of computer-generated random numbers to receive either 5000 iu human chorionic gonadotrophin (hCG) 4 and 8 days after embryo transfer (group 1, n = 534) or 500 mg hydroxy-progesterone caproate and 10 mg oestradiol valerate on days 2, 6, 10 and 14 after embryo transfer (group 2, n = 411), by i.m. injection. Total pregnancy rate was 24% (97 of 411) and 23% (121 of 534), and median serum progesterone concentrations on day 15 after embryo transfer were 5.8 ng ml-1 (range 0.1–298) and 0.1 ng ml-1 (range 0.1–372) in groups 1 and 2, respectively (P = 0.001). One hundred and sixty-three (30.5%) women in group 1 and 22 (5.4%) women in group 2 developed signs of OHSS (P < 0.0001). These results indicate that steroidal ovarian suppression during the luteal phase is a promising tool to reduce the incidence and severity of OHSS in a high-risk population without compromising the pregnancy rate.

Preliminary experience with steroidal ovarian suppression for prevention of severe ovarian hyperstimulation syndrome in IVF patients

The purpose of the study was to evaluate the efficiency of administration of high dose progesterone in combination with oestradiol during the luteal phase for the prevention of ovarian hyperstimulation syndrome in a high-risk population of patients undergoing in vitro fertilization. An observational study was carried out involving 21 women (mean age 28.6 years) undergoing controlled ovarian stimulation for in vitro fertilization. The women were identified as at risk of ovarian hyperstimulation syndrome as they had suffered from the condition when hCG was used for luteal phase support in previous cycles. Steroidal suppression of the stimulated ovary on days 2, 6, 10 and 14 after embryo transfer was achieved by intramuscular injections of 500 mg hydroxyprogesterone caproate and 10 mg oestradiol valerate. The incidence of moderate and severe ovarian hyperstimulation syndrome under steroidal ovarian suppression, serum progesterone concentration and pregnancy rates were compared with those in cycles in which human chorionic gonadotrophin was used for luteal phase support. No cases of moderate or severe ovarian hyperstimulation syndrome occurred under steroidal ovarian suppression. Despite low progesterone concentrations (mean 10.7 nmol l-1, range 2.6–24.5), indicating almost complete ovarian suppression, the pregnancy rate was not impaired. These preliminary results indicate that steroidal ovarian suppression during the luteal phase is a promising tool for reducing the incidence and severity of ovarian hyperstimulation syndrome in a high-risk population, without compromising the pregnancy rate.

Alveolar echinococcosis with bulky peritoneal spread - a rare but important diagnosis in gynaecological practice

A 40 year old woman presented to the outpatient department of our clinic with abdominal distension and a weight gain of 5 kg in the previous three months. Her health was otherwise normal. While she found vaginal examination too uncomfortable to tolerate, a rectal examination was performed which revealed numerous small nodules in the recto-vaginal pouch. Laboratory investigation showed normal CA125 levels, but abdominal ultrasound and computed tomography scan raised the suspicion of a large mass in the lower abdomen, large round lesions in the liver and spleen, and a thickened gastric wall. The small volume of ascites was detected by ultrasound examination but not by computed tomography. Gastroscopy and colonoscopy did not show any pathology, and mammography and a chest X-ray were normal. A diagnostic laparoscopy was performed, which revealed multiple smooth, white peritoneal nodules covering the uterus and bladder (Fig. la). Thick masses in the greater omentum obscured the ovaries. Both the greater omentum and the parietal peritoneum showed thick tumour deposits. Similar findings were seen on the surface of the liver (Fig. lb), the lesser omentum and the spleen. Representative biopsies were taken. Histological examination showed the features of disseminated peritoneal echinococcosis as well as multiple irregular cystic cavities lined with characteristic laminated membranes and filled with amorphous material (Fig. 2). This diagnosis was confirmed by speciesspecific western blots revealing an infection with Echinococcus multilocularis. The woman suffered unexplained post-operative fever for three days. Treatment with albendazole 800 mg per day was started. Three weeks after starting

Evaluation of RU-27987 as a ligand to determine the progesterone receptor

RU-27987, a synthetic progestin, which was recently developed by Roussel Uclaf, Paris, was tested for its validity as a ligand to determine the progesterone receptor in breast cancer. The results were compared to those obtained with R-5020 and ORG-2058, two ligands that are already in use worldwide. The intra- and interassay variation of receptor determination was similar for all 3 ligands. Receptor levels were analyzed with each of the 3 progestins in control cytosols and in 26 mammary carcinoma samples. A good correlation between receptor levels was found although the values of ORG-2058 were somewhat lower, but not significantly. This resulted in a lower proportion of receptor positive samples for ORG-2058 (11/26) compared to R-5020 (13/26) and RU-27987 (14/26). The affinity to the progesterone and to the glucocorticoid receptor, as well as the precision of the Scatchard plot analysis were comparable for the 3 ligands tested. Intra- and interassay variation of receptor determination were also similar. We therefore conclude, that RU-27987 is a suitable ligand to determine progesterone receptor in mammary carcinoma.