Growth-modulatory effects of heparin and VEGF165 on the choriocarcinoma cell-line JEG-3 and its expression of heparanase.

BACKGROUND: Expression of heparanase (HPSE) in tumor cells is strongly associated with invasion, metastasis and angiogenesis. It also plays a key role during pregnancy, in processes of implantation as well as placentation. Vascular endothelial growth factor (VEGF) and heparin are known to alter HPSE expression, with heparin given prophylactically to women with a history of placenta-mediated complications in subsequent pregnancies. MATERIALS AND METHODS: We examined the growth-modulatory effects of different concentrations of heparin and VEGF on the choriocarcinoma cell-line JEG-3 and the expression of heparanase under VEGF and heparin by proliferation assays, PCR, and western blot. RESULTS: Proliferation of JEG-3 cells was induced by heparin in a dose-dependent manner, whereas highly concentrated VEGF led to a decreased cell proliferation. Both agents did not influence the HPSE-expression. CONCLUSION: The presumed pregnancy-protecting effects of heparin may partially be due to an increase of trophoblast proliferation and not via regulation of HPSE expression.

Cancer in pregnancy. Part I: basic diagnostic and therapeutic principles and treatment of gynecological malignancies.

PURPOSE: Cancer in pregnancy is a rare circumstance. However, the coincidence of pregnancy and malignancy is supposed to increase due to a general tendency of postponing childbearing to older age. To date, clinical guidelines are scarce and experience regarding therapeutic management is limited to case reports. METHODS: This review focuses on general diagnostic and therapeutic principles including systemic therapy for malignancies in pregnancy. RESULTS: In part I, we report on diagnosis and therapy of gynecological tumors. CONCLUSION: The diagnosis of gestational cancer faces both oncologist and obstetrician to the dilemma of applying appropriate diagnostic techniques and adequate local and systemic therapy to an expectant mother without harming the fetus.

Cancer in pregnancy. Part II: treatment options of breast and other non-gynecological malignancies.

PURPOSE: In case of non-gynecological solid tumors and hematological malignancies diagnosed during pregnancy, individual diagnostic and treatment options must be established by an interdisciplinary team. METHODS: In part II of the present review we report on diagnostic and therapeutic principles in distinct entities of solid and hematological malignancies. RESULTS: On the basis of a review of the current literature, clinical guidelines and algorithms have been established for diagnosis and therapy of maternal cancer during pregnancy. CONCLUSIONS: The prognosis of the malignancy and the patient's informed consent must be taken into consideration when the well-being of the expectant mother is weighed against the well-being of the unborn child in case of maternal cancer during pregnancy.

Current diagnostic modalities and clinical pitfalls in malignant secondary breast tumours.

Breast cancer is the most common malignancy in women with increasing incidence. The occurrence of metastatic disease to the breast in both females and males is relatively rare, constituting 0.5-6% of all breast malignancies. The therapy of secondary breast cancer is usually completely different from that of primary malignant lesions. Thus, early confirmation of secondary involvement is crucial to direct appropriate and to avoid potentially harmful treatment. The main focus remains to include secondary breast involvement in the differential diagnosis of breast tumours. In recent years, a number of new or improved imaging modalities have been introduced in the diagnosis of breast neoplasm. Current diagnostic concepts of metastatic neoplasm to the breast, including latest imaging modalities and their possible future applications, are presented in this review with special emphasis on possible pitfalls.

Laparoendoscopic single-site surgery (LESS) - is it feasible in gynecological surgery?

Increases in technical expertise in gynecological surgery and advances in surgical instrumentation have led to the development of laparoendoscopic single-site surgery (LESS). Between March and September 2009, 24 patients underwent adnexal surgery at our institution with laparoendoscopic single-site surgery. The LESS technique was performed using the TriPort through an umbilical incision of 10 mm and bent laparoscopic instruments. We furthermore compared the LESS technique with a control group of 24 patients operated consecutively in the same period and for the same procedures with conventional multiport laparoscopy. Comparing the two techniques we found differences between the operation time and mean hospital stay. The surgeon must master the use of novel bent instruments in close proximity to each another. The LESS technique for benign adnexal surgery is technically feasible and safe, representing a reproducible alternative to conventional multiport laparoscopy.

Fetal micrognathia: objective assessment and associated anomalies on prenatal sonogram.

OBJECTIVE: To determine the accuracy and characteristics of prenatally detected fetal micrognathia. METHODS: A retrospective analysis of all pregnancies with the suspicion of fetal micrognathia was performed. The affected fetuses were reassessed by estimation of the inferior facial angle (IFA) and the frontal nasomental angle on stored gray scale images to objectively establish the diagnosis. RESULTS: Of the 28.935 ultrasounds (USs) reviewed, 58 cases were eligible and 4 were excluded because of inconclusive data. The mean values for IFA and frontal nasomental angle were 44.8° and 123.3°, respectively. In 33 cases, the pregnancy was terminated. Four fetuses died sub partu or immediately after birth, five were stillborn. Invasive testing in 40/54 cases revealed aneuploidies in 35%. Associated anomalies comprised musculoskeletal disorders (43%) and non-skeletal anomalies (15%). Less than one fifth (9/54) were alive beyond postnatal period. Four fetuses had an isolated micrognathia, one of which was found to have a cleft palate postnatally. CONCLUSION: The diagnosis of micrognathia has a crucial impact on both prenatal and postnatal outcomes of affected individuals due to its association with additional abnormalities. A detailed sonographic survey using objective criteria for defining micrognathia is mandatory. Once the diagnosis is confirmed, an intensive interdisciplinary counseling of the parents is needed.

Successful application of vacuum-assisted closure therapy for treatment of mastitis-associated chronic breast wounds.

INTRODUCTION: Although non-puerperal mastitis is rare and its cause is rather unclear, the number of patients diagnosed with this defect is increasing. In some cases, standard therapy fails and it progresses to a chronic disease. Vacuum-assisted closure (VAC) therapy has shown good results in healing complex wounds. PATIENTS AND METHODS: The goal of this study was to evaluate our experiences with VAC therapy and to answer the question whether or not should be accepted as an effective treatment in healing chronic wounds caused by non-puerperal mastitis. Retrospectively, we identified five patients with a non-puerperal mastitis chronic wound that was treated using VAC and report on their outcome. RESULTS: We reached both wound control and closure of the breast wounds in all patients. CONCLUSION: According to our findings, the VAC therapy can be considered when managing challenging breast wounds, particularly when other therapeutic options have failed. The role of VAC therapy as a primary therapeutic option has not yet been evaluated.

Heparanase expression in term placentas of diabetic patients and healthy controls.

PURPOSE: The prevalence of diabetic disorders in pregnancy is rising, which goes along with increased risks for maternal and foetal complications during pregnancy and delivery. The expression of the endo-ß-D: -glucuronidase, heparanase (HPSE), may increase under hyperglycaemic conditions, is believed to play an important role in diabetes associated morbidity outside the female reproductive tract and is expressed in the placenta throughout gestation. However, the placental expression of HPSE has not been investigated in diabetic patients. MATERIALS AND METHODS: Placental biopsies of 30 patients with pre-existing or gestational diabetes and 30 healthy controls were analysed by real-time PCR and immunohistochemistry with regard to the presence of HPSE at term. RESULTS: Patients and controls were comparable with respect to foetal outcome and maternal characteristics except for maternal body mass index. We were unable to show significant differences in placental HPSE expression between diabetic patients and healthy controls. DISCUSSION: This study suggests that HPSE expression in term placentas is not affected by maternal diabetes and thus does not contribute to pathological processes in diabetic pregnancies with deliveries at term.

Hysteroscopic findings in women with two and with more than two first-trimester miscarriages are not significantly different.

The purpose of this study was to analyse hysteroscopic results in patients with recurrent miscarriages and to compare the frequency of uterine anomalies in women with a history of exactly two and with more than two consecutive miscarriages. A retrospective analysis of 206 patients undergoing hysteroscopy for repeated early pregnancy losses was performed at two university centres. Late miscarriages were excluded, terminations of pregnancy were not counted. Eighty-seven patients had suffered from exactly two early miscarriages and 119 from more than two. Both groups were comparable with respect to age at admission (32.95+/-4.46 versus 34.06+/-5.02 years) and at first miscarriage (30.43+/-4.24 versus 29.08+/-5.38 years). The prevalence of acquired (adhesions, polyps, fibroids) and congenital uterine anomalies (septate or bicornuate uterus, etc.) did not differ significantly (acquired: 28.7 versus 27.7%; congenital: 9.2 versus 16.8%). The rates of uterine anomalies did not differ significantly overall (36.8 versus 42.9%). In conclusion, uterine anomalies are frequently found in patients with two and with more than two early miscarriages. Due to the high rate of anomalies, their risk for adverse pregnancy outcome and a possible therapeutic approach, hysteroscopy might be a diagnostic option even after two early miscarriages.

Nulliparous patients with small stature delivering at term have an increased risk of secondary cesarean section.

PURPOSE: To evaluate the obstetric outcome of pregnant patients with small stature (<5th percentile) with regard to the mode of delivery, maternal injuries, and neonatal parameters. METHODS: Retrospective cohort analysis of 13 years of deliveries. Two groups: group A, patients with a height below the 5th percentile, and group B, patients with a body height between the 25th and 75th percentile. RESULTS: Patients with a body height between the 25th and 75th percentiles showed significantly more spontaneous vaginal deliveries. Secondary cesarean sections (CS) were significantly seen more often in mothers with a small body height. The fetal outcome did not differ significantly between both groups (APGAR, arterial cord pH, base excess). CONCLUSIONS: Patients with body height below the 5th percentile were found to have a significantly higher rate of secondary CS. As less than half of our patients with a body height below the 5th percentile were found to have delivered spontaneously at term, pregnancies in small patients should be recognized by obstetricians to be at a specific risk. Whereas the neonatal outcome appears to be comparable between nulliparous women with a body height below the 5th percentile and those with a body height between the 25th and 75th percentiles, small mothers carry a significantly elevated risk of surgical delivery, which should be addressed in prospective studies and in counseling these patients.

Advanced age is a risk factor for higher grade perineal lacerations during delivery in nulliparous women.

PURPOSE: To identify risk factors for the development of severe perineal lacerations and to give recommendations for their prevention in nulliparous women. METHODS: A retrospective case-control analysis of deliveries at our University Hospital was performed. Multiparae, Caesarean sections, twin pregnancies, fetal breech position and preterm deliveries were excluded. Univariate and multivariate step forward regression analyses were performed; correlations between contributors were further analyzed by Spearman Rank Correlation. Differences between the degree of lacerations and maternal age were further analyzed with Friedman ANOVA followed by Dunn's Multiple Comparison Test. RESULTS: A total of 2,967 deliveries fitted our inclusion criteria, 50 (1.7%) mothers had higher-grade lacerations. Mediolateral and median episiotomy, advanced maternal age, vaginal operative delivery, higher fetal birth weight and abnormal cephalic presentation were associated with severe lacerations. CONCLUSIONS: Advanced maternal age plays an important role in the development of anal sphincter tears in nulliparous women. Episiotomy and operative vaginal deliveries should be restrictively performed when possible. To identify further preventive approaches in patients with accumulated risk factors prospective randomized studies are needed.

Specific secretory phase endometrial leukocytes of women with two and more consecutive idiopathic abortions are not significantly different from healthy controls.

OBJECTIVE: To analyze concentrations of endometrial leukocytes in patients with idiopathic-repeated abortions. MATERIALS AND METHODS: Biopsies of exactly dated secretory endometrium in 25 patients with idiopathic-repeated abortions and 10 control patients without a history of miscarriage were compared with respect to the concentrations of T-helper cells (CD4), cytotoxic T-cells (CD8), B-cells (CD19) and uterine natural killer cells (CD56) by immunohistochemistry and RNase protection assays. RESULTS: All examined cells were detectable within secretory endometrium. No statistically significant differences of the examined immune-cell concentrations were seen between the control group and the repeated miscarriage group by either test. CONCLUSION: This study suggests that the concentrations of specific endometrial leukocytes in a non-pregnant cycle are not associated with repeated pregnancy loss. Thus, the hypothesis of an altered endometrial immunity in patients with repeated miscarriages, symbolized by persistently differing local immune-cell concentrations, has to be questioned.

Sildenafil inhibits the proliferation of cultured human endothelial cells.

The proliferation of endothelial cells plays a crucial role in the development of intraplaque angiogenesis (IPA). IPA is a major source of intraplaque hemorrhage and therefore contributes to the destabilization of atherosclerotic plaques. Therefore, the aim of the present study was to examine, whether sildenafil inhibits endothelial cell growth. The proliferation of human endothelial cells derived from umbilical cord veins (HUVEC) was examined on DNA level by measurements of ((3)H)-thymidine incorporation. Cell viability was analyzed using trypan blue staining. The proliferation of cultured human endothelial cells was significantly decreased by 1 µmol/l (-48.4%) and 10 µmol/l (-89.6%) sildenafil (n=10, p<0.05). This was not a cytotoxic effect, because cell viability was only reduced at sildenafil concentrations of 50 µmol/l or greater. In addition sildenafil significantly reduced endothelial proliferation induced by bFGF (n=10, p<0.05). The presented results demonstrate an antiangiogenic effect of sildenafil that might be useful in the prevention of atherosclerotic plaque vascularization.

Role of cGMP in sildenafil-induced activation of endothelial Ca2+-activated K+ channels.

Intracellular cGMP is an important second messenger in endothelial cells. Because Ca(2+)-activated K(+) channels with large conductance (BK(Ca)) have been shown to regulate endothelial cell functions, the aim of the present study was to examine whether sildenafil modulates BK(Ca) activity in cultured human endothelial cells. Changes of the endothelial cell membrane potential were analyzed using the fluorescence dye DiBAC. The patch-clamp technique was used to study BK(Ca) in human endothelial cells of umbilical cord veins (HUVEC). Intracellular Ca(2+) levels were analyzed using Fura-2 fluorescence imaging. Sildenafil caused a dose-dependent (0.05-5 micromol/l) hyperpolarization of the endothelial cells with a maximum at a concentration of 1 micromol/l. A significant increase of BK(Ca) activity was induced by sildenafil (1 micromol/l) perfusion. BK(Ca) open state-probability (NPo) was also increased by the cGMP-analogue 8-bromo-cGMP (0.5 mmol/l), whereas inhibition of the cGMP-dependent kinase (PKG) had no effect on NPo. PKG-inhibition abolished 8-bromo-cGMP induced BK(Ca) activation, and reduced sildenafil induced NPo. Furthermore, sildenafil caused a significant increase of intracellular calcium that was blocked by the BK(Ca) inhibitor iberiotoxin (100 nmol/l). In conclusion sildenafil activates BK(Ca) by a mechanism, which involves cGMP. The activation of the BK(Ca) is responsible for the sildenafil-induced increase of intracellular Ca(2+).

Margatoxin inhibits VEGF-induced hyperpolarization, proliferation and nitric oxide production of human endothelial cells.

BACKGROUND: Vascular endothelial growth factor (VEGF) induces proliferation of endothelial cells (EC) in vitro and angiogenesis in vivo. Furthermore, a role of VEGF in K(+) channel, nitric oxide (NO) and Ca(2+) signaling was reported. We examined whether the K(+) channel blocker margatoxin (MTX) influences VEGF-induced signaling in human EC. METHODS: Fluorescence imaging was used to analyze changes in the membrane potential (DiBAC), intracellular Ca(2+) (FURA-2) and NO (DAF) levels in cultured human EC derived from human umbilical vein EC (HUVEC). Proliferation of HUVEC was examined by cell counts (CC) and [(3)H]-thymidine incorporation (TI). RESULTS: VEGF (5--50 ng/ml) caused a dose-dependent hyperpolarization of EC, with a maximum at 30 ng/ml (n=30, p<0.05). This effect was completely blocked by MTX (5 micromol/l). VEGF caused an increase in transmembrane Ca(2+) influx (n=30, p<0.05) that was sensitive to MTX and the blocker of transmembrane Ca(2+) entry 2-aminoethoxydiphenyl borate (APB, 100 micromol/l). VEGF-induced NO production was significantly reduced by MTX, APB and a reduction in extracellular Ca(2+) (n=30, p<0.05). HUVEC proliferation, examined by CC and TI, was significantly increased by VEGF and inhibited by MTX (CC: -58%, TI --121%); APB (CC --99%, TI--187%); N-monomethyl-L-arginine (300 micromol/l: CC: -86%, TI --164%). CONCLUSIONS: VEGF caused an MTX-sensitive hyperpolarization which results in an increased transmembrane Ca(2+) entry that is responsible for the effects on endothelial proliferation and NO production.