Transforming growth factor-beta signaling leads to uPA/PAI-1 activation and metastasis: a study on human breast cancer tissues.

Metastasis represents a major problem in the treatment of patients with advanced primary breast cancer. Both Transforming Growth Factor-Beta (TGF-ß) signaling and Plasminogen Activator (PA) components, urokinase-type Plasminogen Activator (uPA) and Plasminogen Activator Inhibitor-1 (PAI-1) represent a complex network crucial for such enhanced invasiveness of tumors and imply high prognostic/predictive and promising therapeutic potential. Therefore, protein expression of specific effector molecules comprising the main parts of the TGF-ß signaling pathway were determined in HOPE-fixed human tumor tissues through IHC (Scoring) using tissue microarray (TMA) technique and correlated with respective uPA and PAI-1 levels determined earlier in the same TMAs through optimized IHC and semi-quantitative image analysis. TGF-ß signaling was active in vast majority (96 %) of the tumor samples and 88 % of all cases were significantly correlated with established metastasis markers uPA and PAI-1. In addition, TGF-ß was also closely associated with tumor size, nodal status and two steroid hormone receptors. Consistent interrelationships between TGF-ß, PA components and additional tumor characteristics underline the superiority of such more comprising data with regards to confirming TGF-ß signaling as a promising target system to inhibit metastasis in advanced breast cancer.

Optimized immunohistochemistry in combination with image analysis: a reliable alternative to quantitative ELISA determination of uPA and PAI-1 for routine risk group discrimination in breast cancer.

The determination of the invasion markers urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) has further improved the possibilities for individualized therapy of breast cancer. To date, quantitative measurement by ELISA, that needs large amounts of fresh, frozen material, is the only standardized procedure for diagnostic purposes. Therefore, the aim of this study was the establishment of a reliable alternative method based on immunohistochemistry (IHC) and image analysis requiring only small amounts of fixed tumor tissue. Protein expression of uPA and PAI-1 was analyzed in HOPE-fixed tumor samples using tissue microarrays (TMAs) and semiquantitative image analysis. The results of both methods were significantly correlated and risk assessment showed an overall concordance of 78% (83/107; high- and low-risk) and of 94% (74/79) regarding only high-risk patients. The data demonstrate that optimized IHC in combination with image analysis can provide adequate clinical significance compared to ELISA-derived determination of uPA and PAI-1.

Apparent up-regulation of stimulatory G-protein alpha subunits in the pregnant human myometrium is mimicked by elevated smoothelin expression.

Sensitization of adenylyl cyclase (AC) by increased expression of large isoforms of the stimulatory G-protein Galpha(s) has been suggested as a mechanism that governs uterine quiescence during pregnancy. We quantified several components of the AC pathway in pregnant (P, n=21) and nonpregnant human myometria (NP, n=10). AC activity was approximately sevenfold higher in P than in NP under basal and stimulated conditions (MnCl(2)/GTP/GTP + isoproterenol). In addition, relative stimulation (% of basal) by 5'-guanosine-betagamma-iminotriphosphate and forskolin was twofold higher in P. beta-Adrenoceptor density was low and unaltered in P. Galpha(s) mRNA splice variants did not differ in P. Using antisera against different epitopes of Galpha(s) (carboxyl-/more amino-terminal), we found unchanged expression of Galpha(s) short and long (45, 47 kDa) in P. Two additional proteins in P (51, 59 kDa) were detectable only by the carboxyl-terminal antiserum and lacked GTP binding properties. The 59 kDa protein could be identified as a recently discovered cytoskeletal protein, smoothelin, which was 10-fold increased in P. These data indicate that the apparent up-regulation of large Galpha(s) species in P is mimicked by elevated smoothelin. Therefore, the increase in AC cannot be attributed to changes in Galpha(s)- or beta-adrenoreceptors. Epitope sharing between Galpha(s) and smoothelin should be considered in experiments on smooth muscle tissues.

[Efficacy of ultrasound screening in pregnancy]. / Qualität des Ultraschall-Screenings in der Schwangerenvorsorge.

OBJECTIVE: The purpose of this study was to examine the effectiveness of ultrasound screening in pregnancy. MATERIAL AND METHODS: Therefore, it was registered whether fetal malformations in a study population of 11,172 deliveries were already diagnosed before birth. RESULTS: 341 defects were found in 297 children from mothers who had had prenatal care. Most anomalies were seen in the urogenital tract (n = 98; 28.7%), the heart (n = 67; 19.6%), the connective tissue (n = 39; 11.4%), the gastrointestinal tract (n = 32; 9.4%), and in the central nervous system (n = 33; 9.7%). Chromosomal anomalies (n = 22; 6.5%) and orofacial defects (n = 21; 6.2%) were more rare. 8.8% of all defects were lethal, 37% severe. 237 (69.5%) were classified as "diagnosable by ultrasound prenatally". 125 of them (53%) were identified prenatally, with high rates of 71% in central nervous system, 65.5% in intestinal and 54% in urogenital tract, while the detection rate was only 13.6% in chromosomal and 3.3% in cardiac defects. Only 14.3% were found before 24 weeks of gestation. CONCLUSIONS: Thus, the effectiveness of ultrasound screening has to be improved by adequate measures.

Oxytocin and vasopressin 1a receptor gene expression in the cycling or pregnant human uterus.

OBJECTIVE: Our purpose was to compare concentrations of messenger ribonucleic acid specific for the oxytocin receptor and for the vasopressin 1a receptor in myometrial and endometrial tissues of pregnant and nonpregnant women. STUDY DESIGN: Tissues from pregnant uteri were obtained from 95 women who were undergoing cesarean delivery between 26 and 42 weeks' gestation. Tissues from nonpregnant uteri were obtained from 7 cycling women who were undergoing hysterectomy. The competitive reverse-transcription polymerase chain reaction method was used to determine messenger ribonucleic acid concentrations. RESULTS: A significant increase in oxytocin receptor messenger ribonucleic acid was found during the first half of pregnancy. Oxytocin receptor messenger ribonucleic acid concentrations were lower in tissues with spontaneous contractions than in quiescent tissues and were decreased in patients with advanced labor. Vasopressin 1a receptor messenger ribonucleic acid concentrations were high in tissues from both cycling and pregnant uteri but remained unchanged throughout gestation. CONCLUSION: The increase in oxytocin receptor protein concentrations seen in pregnancy is only partially controlled by messenger ribonucleic acid abundance. High concentrations of vasopressin 1a receptor messenger ribonucleic acid confirm the biologically active role of this receptor in both the cycling and the pregnant uterus.

Oxytocin and vasopressin receptors in human and uterine myomas during menstrual cycle and early pregnancy.

The purpose of this study was to determine the specificity and concentration of oxytocin (OT) and arginine vasopressin (AVP) binding sites in non-pregnant (NP) human and rhesus monkey endometrium, myometrium and fibromyomas, and to determine the cellular localization of OT receptor (OTR). Besides [3H]AVP, [125I]LVA, a specific VP1 receptor subtype antagonist, was used to determine vasopressin receptor (VPR) concentrations. Samples were obtained from 42 pre-menopausal and three pregnant women (5, 13 and 35 weeks gestation), and several NP and pregnant monkeys. Specificity of binding was assessed in competition experiments with unlabelled agonists and antagonists of known pharmacological potency. Cellular localization of OTR was determined by immunohistochemistry. In NP human uterine tissues, [3H]AVP was bound with higher affinity and greater binding capacity than [3H]OT, whereas in pregnant women and in NP and pregnant rhesus monkeys, uterine OT binding capacity was greater. OT and AVP binding sites discriminated very poorly between OT and AVP; [125I]LVA binding sites were more selective than [3H]AVP. Their ligand specificity and binding kinetics indicated the presence of two distinct populations of binding sites for OT and AVP in primate uterus. Endometrium of NP women and monkeys had low OTR and VPR concentrations. Myometrial and endometrial OTR and VPR were down-regulated in midcycle and in early human pregnancy, they were up-regulated in the secretory phase and second half of pregnancy. Immunoreactive OTR in NP uterus was localized in patches of myometrial muscle cells and small numbers of endometrial epithelial cells.

[Rhesus prophylaxis--history and current status]. / Die Rhesusprophylaxe--Geschichte und aktueller Stand.

The varying symptoms of rhesus incompatibility were recognized and described approximately sixty years ago. Jörg Schneider, then at the Freiburg University Hospital in Germany, had been the very first investigator to perform rhesus prophylaxis in pregnant women. The exact date of this achievement is August 9, 1963. One year later, Schneider could report on nine women, who-following delivery of a rhesus-positive child-did not develop rhesus antibodies during a subsequent pregnancy with a rhesus-positive fetus. Since the late sixties, rhesus prophylaxis has been an integral part of prenatal care in Germany. In the meantime, the role of ante-partum prophylaxis has also been established. Since the introduction of post-partum rhesus prophylaxis, fetal morbidity and mortality has been reduced by 90 percent.

[Prognostic value of cardiotocography in pregnancy with pathologic Doppler findings]. / Prognostischer Wert der Kardiotokographie bei Schwangerschaften mit pathologischem Dopplerbefund.

Absent (absent-EDF) or reversed (RF) end-diastolic flow in Doppler velocimetry in the umbilical artery and fetal aorta indicates highly disturbed fetoplacental perfusion, which often occurs early in pregnancy. Perinatal mortality and morbidity is very high. In this situation it is not clear how long pregnancy can be continued to achieve better conditions for the preterm fetus. This study was performed to investigate the value of fetal heart rate recordings (FHR) in patients with absent EDF or RF as a parameter to determine the optimum time of delivery. 25 pregnancies with absent (n = 21) or reversed end-diastolic velocity (n = 4) were analysed. Gestational age was between 24 and 34 weeks. 17 children showed compensatory dilatation of the middle cerebral artery (68%). 15 (60%) were growth retarded, 8 of them below the 3% percentile, while 3 had normal weight. Maximum time interval until delivery was 18 days, median was 2 days. Five children died before, 2 during delivery. As none died after delivery, perinatal mortality was 28%. 4 of the survivers (22%) showed arterial acidosis. Only three children had completely normal fetal heart rate recordings on the day of diagnosis of absent EDF or RF. All others showed at least suspicious FHR (according to Hammacher), 8 of them (32%) even pathological FHR, including all children who died later. Early suspicious changes in FHR were the reduction in frequency and amplitude of oscillation, while decelerations occurred later. Additional unfavourable parameters in FHR were roundings of the oscillatory inversions as well as missing reactivity to fetal movements. Fetal heart rate monitoring is a reliable tool for determination of the optimum time of delivery in children with absent EDF or RF. The decision has to be made in relation to the gestational age. If possible, the fetus should be born before FHR becomes pathological. In this manner perinatal mortality and morbidity should be kept at a low level even in high-risk patients with absent EDF or RF.

[Induced labor with prostaglandin E2 gel after previous cesarean section]. / Geburtseinleitung mit Prostaglandin-E2-Gel bei Zustand nach Sectio.

With the rising rate of Caesarean sections, the number of pregnant patients with a previous Caesarean is increasing. Taking into consideration certain contraindications, it seems to be justified, to attempt induction of labour for vaginal delivery without major risk. However, the use of prostaglandins for cervical ripening in women with an unfavourable cervical status is particularly controversial. Thus, we analysed data from 385 trials of vaginal labour induction in a total of 522 patients with previous Caesarean section. Single or multiple cervical doses of prostaglandin-E2-gel had to be administered because of an unripe cervix (Bishop-score < 8) in 161 women for induction of labour for medical indications. 84.9% of those patients, in which labour induction was attempted after previous Caesarean section, delivered vaginally; 70% after two Caesareans. The highest success rates were seen after previous Caesarean for breech presentation, while there was still a vaginal delivery rate above 60% even after Caesarean for cephalopelvic disproportion or failure to progress. Maternal and foetal complications were seen with equal incidence in both study groups. Uterine ruptures totalling 0.5% were seen only in patients without cervical priming. Taking into consideration contraindications, intracervical application of PGE2-gel is a safe and effective method, even in patients after previous Caesarean section with clear advantages in case of an unripe cervix.

[Antibody detection after antepartal rhesus prophylaxis: normal values or sensitization]. / Antikörper-Nachweis nach antepartaler Rhesus-Prophylaxe: Normalfall oder Sensibilisierung?

Antibody screening tests were performed in 29 unsensitized pregnant women after antepartum Rh immune prophylaxis, using the indirect Coombs test (ICT) and a more sensitive ID-microtyping-system (IDM). With the ICT, anti-D antibodies were detected in 85% for at least 4 weeks and at most 8 weeks after immunisation. The maximum titer was 1:8. With the IDM, 97% showed antibodies against 'D' for at least 4 weeks and at most 11 weeks with a maximum of 1:16. The IDM titer was always 1 to 3 steps more sensitive than the ICT. After postpartum Rh immune prophylaxis, anti-D titers were again positive in many of the patients (ICT: 42%; IDM: 60%). In conclusion, it is nearly always possible to measure antibodies against 'D' after antepartum Rh immune prophylaxis and IDM was superior in comparison to ICT. However, maternal isoimmunisation to the rhesus antigen cannot be excluded for sure and patients have then to be controlled. As isoimmunisation could not be confirmed in any of our patients, postpartum Rh immune prophylaxis has to be administered even after detection of an antibody titer against 'D' after antepartum Rh prophylaxis.

Effects of lumbar epidural analgesia on prostaglandin F2 alpha release and oxytocin secretion during labor.

The concentrations of plasma oxytocin and prostaglandin F2 alpha metabolite (PGFM) were measured in 10 parturients with and 10 without lumbar epidural analgesia. A blood sample was taken immediately before analgesia and another 60 min later. The control patients were matched for the stage of cervical dilatation at the time of the first blood sample; the second was drawn 60 min later. Plasma PGFM decreased significantly after lumbar epidural anesthesia and increased in controls resulting in a highly significant difference between the groups (P < 0.005). Plasma oxytocin concentrations levels also changed in opposite directions in the two groups but the difference did not reach statistical significance (P < 0.1). Uterine activity increased in the controls and decreased in the analgesia group resulting in a significant difference between the groups (P < 0.05). All subjects delivered vaginally. The total duration of labor was longer in the analgesia group (7.8 +/- 1.0 h vs. 4.7 +/- 0.6 h; P < 0.05) as was the duration after analgesia (5.1 +/- 0.9 h vs. 2.5 +/- 0.8 h; P < 0.05), whereas the duration of the second stage was not significantly different. We conclude that lumbar epidural anesthesia results in suppression of PGF2 alpha release which may be the cause of the diminished uterine activity and the prolonged duration of the first stage of labor.

Plasma prostacyclin and thromboxane concentrations in 160 normotensive, hypotensive, and preeclamptic patients during pregnancy, delivery, and the post partum period.

Plasma concentrations of 6-keto-prostaglandin F1 alpha, a stable metabolite of prostacyclin, and TxB2 were measured in 160 women during pregnancy (n = 106), delivery (n = 40), and in the postpartum period (n = 14). Fifty nine patients had normal blood pressure, 10 had mild and 9 severe preeclampsia while 38 patients were hypotensive. Normotensive patients were grouped according to their gestational age: 22-26 weeks (n = 22), 27-31 weeks (n = 22), and 32-38 weeks (n = 15). 20 patients were in early first stage of delivery (cervical dilatation < or = 5 cm), 20 patients in late first stage (cervical dilatation > or = 6 cm). The concentration (mean value +/- SEM) of the PGI2 metabolite tended to increase during pregnancy without reaching significance (218 +/- 11; 225 +/- 10; 250 +/- 15 pg/ml). At the same time, TxB2 showed a decrease, which was most pronounced at 27-31 weeks (65 +/- 15; 40 +/- 2; 48 +/- 4 pg/ml; p < 0.001). The ratio of PGI2/TxA2 increased in parallel (4.9 +/- 0.4; 6 +/- 0.4; 4 +/- 0.5). There was no difference in plasma concentrations of PGI2 (figure 4) and TxA2 in patients with normal blood pressure, mild preeclampsia and hypotension, whereas in severe preeclampsia, the plasma concentration of PGI2 was significantly lower (p < 0.001) and of TxA2 significantly higher (p < 0.001). The ratio of PGI2/TxA2 shifted significantly to vasoconstriction in patients with severe preeclampsia (p < 0.0001) and to vasodilatation in those with hypotension (p < 0.03).(ABSTRACT TRUNCATED AT 250 WORDS)

Oxytocin and bovine parturition: a steep rise in endometrial oxytocin receptors precedes onset of labor.

Oxytocin receptors were measured in myometrium and intercaruncular endometrium of cows during pregnancy and parturition. Concentrations of estradiol-17 beta, estrone, and progesterone in peripheral blood were also measured. Receptor concentrations in the endometrium rose almost 200-fold from Day 20 to term (p < 0.0001, ANOVA), from 40 +/- 11 to 7300 +/- 1430 fmol/mg protein. Myometrial receptor concentrations increased 10-fold from 180 +/- 36 fmol/mg on Day 20 to 1850 +/- 360 fmol/mg protein at term (p < 0.0001, ANOVA). During labor, endometrial receptors (6600 +/- 1300 fmol/mg) remained at prelabor values, whereas myometrial receptor concentrations had decreased to 1190 +/- 316 fmol/mg (not significant) and declined further postpartum. Plasma concentrations of progesterone declined from 4-5 ng/ml to about 2 ng/ml between Days 250 and 282 and dropped to < 0.2 ng/ml shortly before delivery. Plasma concentrations of estrone and estradiol-17 beta were below 10-20 pg/ml until Day 230. Estrone concentrations were significantly (p < 0.05) increased by Day 250 and estradiol-17 beta by Day 270, and then both rose rapidly. During labor, plasma estrone was 1135 +/- 245 pg/ml and plasma estradiol-17 beta was 226 +/- 131 pg/ml. The molar ratio of estrone and estradiol-17 beta to progesterone rose from less than 0.01 to 4.4 during labor, and was correlated with oxytocin receptor concentrations in endometrium (r = 0.5160, p < 0.001), but not those in myometrium (r = 0.0122). The regulation of oxytocin receptors by ovarian hormones in the two tissues may therefore differ.(ABSTRACT TRUNCATED AT 250 WORDS)

Correlation of nocturnal increase in plasma oxytocin with a decrease in plasma estradiol/progesterone ratio in late pregnancy.

OBJECTIVES: Our purpose was to determine whether plasma oxytocin concentrations show a daily rhythm. STUDY DESIGN: Ten women between 37 and 39 weeks of gestation volunteered for the study. They were admitted 1 to 2 days before the experiment. Three blood samples were taken with 2-minute intervals each time, at 8 AM, 4 PM, and 12 midnight. Oxytocin, 17 beta-estradiol, progesterone, and cortisol were measured by radioimmunoassay with highly specific antibodies. Statistical analysis of variance by Friedman's test was followed by multiple range testing, and p < 0.05 was considered significant. RESULTS: Significant daily rhythm was found for plasma cortisol, progesterone, and oxytocin and for the estradiol/progesterone ratio. Oxytocin showed a nocturnal peak and a strong negative correlation with the estradiol/progesterone ratio. CONCLUSIONS: The daily rhythm in plasma oxytocin parallels the rhythm in uterine activity (shown by others), suggesting a causal relationship between the two. Both may in turn be related to the ratio of circulating estradiol and progesterone.

[Doppler ultrasound in monitoring twin pregnancies with early discordant growth]. / Dopplersonographie zur Uberwachung von Zwillingsschwangerschaften mit frühzeitigem diskordanten Wachstum.

Early growth retardation in one twin is complicated by a high fetal mortality and morbidity. It is often very difficult to determine the optimum time of delivery in regard to placental insufficiency and prematurity. We report of two cases in which doppler velocimetry was a helpful method in monitoring those pregnancies at high risk. A growth retardation of three weeks was found in one child of a twin pregnancy at 17 weeks of gestation. Doppler velocimetry at 23 weeks showed a reverse flow and centralisation and the child died at 27 weeks. The other twin then also developed pathologic flow patterns and had to be delivered by cesarean section. In the second patient, pregnancy could be continued in spite of a growth retardation of three weeks in one child at 15 weeks and an additional decrease in the growth rate of both children from 30 weeks onwards. Cesarean section had to be performed at 34 weeks because of premature rupture of membranes.

[Pregnancy follow-up of a twin pregnancy with acardius acephalus]. / Schwangerschaftsverlauf einer Geminigravidität mit Acardius acephalus.

Beginning with the 20th week of gestation a monoamniotic twin pregnancy with acardius acephalus was intensively observed by ultrasound and doppler ultrasound. The acardius only grew until the 24th week of gestation; the fetus underwent a normal development during the entire pregnancy. Doppler ultrasound showed low resistance indices in both umbilical arteries. During the 24th week of gestation the flow in the umbilical artery of the acardius could no longer be registered due to a cessation of development. A normal flow could be registered from the 28th week of gestation until the end of pregnancy in the umbilical artery, aorta fetalis and the art. cerebri med. of the fetus. During the 36th week of gestation the fetus died because of umbilical cord entanglement with the cord of the acardius.