Characterization of 17beta-hydroxysteroid dehydrogenase type 7 in reproductive tissues of the marmoset monkey.

In contrast to the known rodent enzymes, the physiological significance of 17beta-hydroxysteroid dehydrogenase type 7 (17HSD7) and its presumed function in reproductive biology is not well understood in primates. As a first step, we recently cloned the complete coding regions of human and marmoset monkey (Callithrix jacchus) 17HSD7 (cj17HSD7). In the present work the complete cDNA of marmoset 17HSD1 (cj17HSD1), including the proximal promoter region, and a partial sequence of marmoset aromatase (cjARO) were sequenced in order to compare the expression of these estradiol synthesizing enzymes with that of 17HSD7 in a primate model and to identify tissues where 17HSD7 might participate in the pathway of estradiol synthesis. The gene structures of cj17HSD1 and cj17HSD7 were determined and proved to be very similar to the human orthologues. Northern hybridization showed that cjARO mRNA seems to be coexpressed preferably with cj17HSD1 in placenta, whereas in other tissues it is expressed in parallel only with cj17HSD7. Especially in corpora lutea, the cj17HSD7 transcript is detectable throughout the luteal phase of the ovarian cycle and increases during pregnancy, in parallel with the transcript of aromatase. Results were confirmed by immunoblots and immunohistochemistry using new polyclonal antisera directed against cj17HSD7 and cjARO protein. The enzymatic conversion of estrone to estradiol was assessed in marmoset corpora lutea. The pattern of coexpression with aromatase supports the hypothesis that luteal 17HSD7 complements placental 17HSD1, ensuring continued estradiol synthesis throughout pregnancy in primates.

[Relaxin: an important indicator of canine pregnancy]. / Relaxin: ein wichtiger Graviditätsindikator bei der Hündin.

In the present study the concentration of relaxin in peripheral blood plasma was assessed during canine pregnancy for its suitability as a pregnancy indicator, using a newly developed relaxin enzyme immunoassay. A significant relaxin increase was found in pregnancy at day 24 after ovulation. However, this relaxin increase did not correlate either with litter size or with body weight of the bitch. Induction of abortion with prostaglandin F2 alpha resulted in reduced peripheral relaxin levels, suggesting a damage of the placenta due to this medical intervention. Thus, the results confirm that relaxin, which is produced by the placenta, is a useful marker for early pregnancy diagnosis in the bitch. Relaxin measurement is recommended for detection of pregnancy either alone, or as supplement of ultrasonographic findings.

Characterization of relaxin binding in the uterus of the marmoset monkey.

The ovarian peptide hormone relaxin (RLX) plays an important role in the regulation of the endometrium both during the cycle and in early pregnancy. RLX interacts with specific receptors on endometrial stromal cells causing these to decidualize. In order to characterize the molecules with which RLX interacts in the primate uterus, a methodology based on a fully bioactive preparation of biotinylated porcine RLX was applied to cryosections of the uterus of female marmoset monkeys. Specific RLX binding was weakly detected in the proliferative phase in isolated endometrial stromal cells. In the secretory phase, the positively reacting cells increased in staining intensity and in number and also included some epithelial cells. Further increases occurred in pregnancy, but RLX binding in the endometrium decreased at the end of the cycle if pregnancy did not occur. The myometrium showed weak staining which did not vary through the cycle, but increased in pregnancy. Electrophoretic analysis of the RLX-binding moieties in these tissue sections indicated that a protein of approximately 40 kDa was the principal RLX-binding molecule, while minor specific bands were detectable at approximately 100 and approximately 200 kDa. The binding of biotinylated RLX could be specifically suppressed by co-incubation with unlabelled RLX, but not by insulin, IGF-I or biotin. This technique therefore allows the detection and molecular characterization of specific RLX binding in the primate uterus. In the marmoset monkey, the pattern of specific binding closely reflects the RLX-dependent physiology during implantation and early pregnancy, implying the probable involvement of a specific RLX receptor.

Relaxin in the marmoset monkey: secretion pattern in the ovarian cycle and early pregnancy.

Relaxin is a peptide hormone with a broad range of biological activities, related not only to parturition and lactation but possibly also to decidualization, implantation, and early pregnancy. The present study was designed to investigate the secretion pattern of relaxin throughout the cycle and early pregnancy in the common marmoset monkey in relation to ovarian function and the systemic hormone milieu. First, a novel relaxin ELISA was developed and validated to confirm the pattern of relaxin secretion during pregnancy. Secondly, serum relaxin profiles were determined through nonconceptive and conceptive cycles and analyzed in relation to the concentration of other hormones and to the development of ovarian follicles and corpora lutea (CL). Blood samples were collected 2-3 times per week from the experimental animals and analyzed for relaxin, progesterone, and LH. The animals from the conceptive cycles were also ultrascanned at these time points to determine the ovarian status up to Day 25 of pregnancy. During early pregnancy, the relaxin levels in serum were approximately 1 ng/ml, increasing up to 15 ng/ml in the second trimester, at a time when progesterone levels had declined. In the third trimester, when progesterone levels were increasing again, the levels of relaxin decreased, returning to basal levels by term of pregnancy. In early pregnancy there was a parallel increase in both relaxin and LH/hCG, with the relaxin rise in the conceptive cycle appearing sooner than in the nonconceptive cycle, suggesting that, like chorionic gonadotropin (CG), relaxin may be a useful and early marker for pregnancy. Unlike the situation in the human, there was no correlation between the levels of either hormone and the number of CL detected, infants born, mother's age, or parity. Relaxin levels increased in early pregnancy before bioactive LH/CG, implying that relaxin is not directly regulated by this gonadotropin. Furthermore, hCG applied to nonconceptive females during the expected time of implantation caused an increase in progesterone but not in relaxin concentrations. In summary, the results obtained indicate that relaxin may be a reliable indicator of early pregnancy status in the common marmoset, but it is independent of direct CG influence.

Local relaxin biosynthesis in the ovary and uterus through the oestrous cycle and early pregnancy in the female marmoset monkey (Callithrix jacchus).

The pattern of peripheral serum concentration for the peptide hormone relaxin in women points to the possibility of an interesting paracrine function in the cycle and early pregnancy. In order to investigate this physiology in detail, it was decided to examine local relaxin biosynthesis in an established primate model for human female reproductive function, the marmoset monkey (Callithrix jacchus). In this initial study relaxin biosynthesis was assessed using a combination of molecular and immunological techniques through the oestrous cycle in the marmoset monkey. The nucleotide sequence of the full-length relaxin gene transcript was cloned from the marmoset ovary and found to be closely homologous to that of the human H2 relaxin. Using gene specific probes derived from this sequence, RNase protection assays, reverse transcription-polymerase chain reaction (RT-PCR) assays and in-situ hybridization, showed relaxin gene expression within the ovary in theca cells and corpora lutea in the oestrous cycle, increasing in early pregnancy. Relaxin gene expression was also identified at a low level in the uterus and placenta, and at a higher level in the prostate in the male marmoset monkey. Using two different relaxin-specific antisera, relaxin-like immunoreactivity was observed in the ovary with a pattern of distribution coincident with that obtained by in-situ hybridization. Immunoreactivity was also found in the non-pregnant uterus, within the endometrial epithelium of the late proliferative phase and increasing within the glands through the secretory phase. Taken together, the pattern of relaxin peptide and mRNA expression show there is the basis for local relaxin physiology within the ovarian follicle and corpus luteum, and within the uterus during the oestrous cycle in this new world monkey.

[Collagenous sprue]. / Kollagene Sprue.

HISTORY AND FINDINGS: A 45-year-old man with type I diabetes mellitus was admitted to hospital because of colicky abdominal pain and 5-6 watery stools daily. Upper gastrointestinal endoscopy showed nearly total atrophy of the villi in the duodenum and jejunum suggesting coeliac disease. However, gluten-free diet for 2 weeks brought no improvement. Another examination of the biopsy 6 weeks after the first endoscopy revealed extensive collagen deposition in the lamina propria of the small intestine, giving the diagnosis of collagenous sprue. TREATMENT AND COURSE: Parenteral nutrition, lactulose, cisapride, cholestyramine, doxycycline, paromomycin, vancomycin and octreotide failed to affect the loss of fluid from the gut which 12 weeks after admission had increased to 221 daily. However, it was stopped after prednisolone was administered (100mg daily). 7 months after starting the steroid treatment the collagen layer had disappeared and the villous atrophy had partially regressed. Over the next 6 months the prednisolone dosage was decreased to 10 mg daily. Shortly thereafter a perimembranous glomerulonephritis occurred, with proteinuria (up to 60 g/d) and oedema. It regressed to 6 g/d when the steroid dose was increased and cyclosporin, 0.5 g/d, had been added. On maintenance dosage of cyclosporin the histological and clinical remission of the collagenous sprue has now lasted for over 2 years. CONCLUSIONS: This case suggests that steroid administration is an effective treatment of collagenous sprue. The presence of diabetes and other immune-related diseases in this case also suggests that an immunological mechanism may play a causative role in collagenous sprue.

Maternal milk and plasma glucose and insulin levels: studies in normal and diabetic subjects.

The relationship between maternal glucose levels and the concentration of glucose and insulin levels in human milk from diabetic women has not been elucidated. In addition, the rate of appearance of intravenously injected insulin to the change in concentration of insulin in maternal milk has not been studied. To study this relationship of glucose levels in serum to glucose levels in milk, maternal milk and glucose levels were measured in diabetic lactating women (n = 7) and nondiabetic lactating women (n = 10). In addition, the change in milk concentration of insulin was studied after an intravenous injection of insulin. The maternal whole blood glucose in the seven diabetic women was stabilized at a baseline blood glucose of approximately 100 mg/dl and then elevated with an infusion of intravenous glucose to a level of three times baseline (approximately 300 mg/dl for up to 2 hours). The plasma glucose was then lowered back to baseline with intravenous insulin over 20 minutes. The baseline serum insulin and glucose levels were compared to nonlactating women who donated serum to measure insulin levels in normal controls. Maternal milk glucose levels rise following an increase of plasma glucose levels with a lag time to the peak glucose level of 40-90 minutes, and return to baseline following the return of plasma glucose to baseline with a lag time of 120-150 minutes. Baseline milk insulin levels are elevated in hyperinsulinemic women and the levels of insulin in the milk will rise dramatically above baseline values after an intravenous injection of insulin with a lag time to the peak of concentration in milk of 60-80 minutes.(ABSTRACT TRUNCATED AT 250 WORDS)

[Rare coincidence of Ehlers-Danlos syndrome, diabetes mellitus and pregnancy]. / Seltenes Zusammentreffen von Ehlers-Danlos-Syndrom, Diabetes mellitus und Schwangerschaft.

A report is given about a seldom coincidence of Ehlers-Danlos-Syndrome, diabetes mellitus and pregnancy in a 31 year old patient.--The course of pregnancy was uncomplicated except a cervical insufficiency and a secondary wound healing of the episiotomy. The exact diagnosis of the Ehlers-Danlos-syndrome should be done already before pregnancy.

Treatment of pregnant insulin-dependent diabetic women.

The treatment of 620 insulin-dependent diabetic pregnant women is reported. The goal of treatment was to achieve a normal blood glucose concentration as soon as possibly during early, or even before pregnancy. When intensified conventional insulin therapy was started before conception, about 88% of the patients achieved normal blood glucose levels during the first weeks of pregnancy. In only about 20% of the pregnant diabetics without intensified preconceptional treatment a normal blood glucose level was obtained during their first hospitalization in pregnancy. The rate of congenital malformations was 1.1% in the former and 7.1% in the latter group.

[Measurement of uteroplacental circulation with 113m indium in diabetic pregnancy]. / Die Messung der uteroplazentaren Durchblutung mit 113m In in der diabetischen Schwangerschaft.

In 122 diabetic pregnancies the placental blood flow has been estimated determining the half life of the activity inflow (2 MBq 113 m In labelled transferrin) into the placental bed. We used a highly sensitive detector (modified pinhole collimator) and a computer supported evaluation, free from subjective influences. 259 flow measurements were compared to the risk of complication in the course of a diabetic pregnancy. - The half life values in the diabetic group, calculated by a gamma camera computer system by means of an iterative regression analysis, were significantly different compared to a control group (12 pregnancies without risk.) - Severe diabetic angiopathic complications (White classes D, F, and R) are accompanied by higher half life values (placental blood flow reductions) and perinatal complications. - Even in pregnant women with gestational diabetes or disturbances of the carbohydrate metabolism a disturbed placental hemodynamic is to be found.

[Relation of uteroplacental circulation and morphologic findings in myometrial and decidual arteries in diabetic pregnancy]. / Uber den Zusammenhang von uteroplazentarer Durchblutung und morphologischen Befunden an den myometrialen und dezidualen Arterien in der diabetischen Schwangerschaft.

Placental blood flow measurements are compared to morphologic blood vessel examinations of the placental bed and uterus. In 122 diabetic pregnant women half life time of 113m Indium labelled blood inflow curves into the placenta as measured by a gamma scintillation camera on-line recorded to a computer system. - In 57 patients histological examination of uteroplacental arteries were done. - Congruent results of placental blood flow and arterial morphology were found in 66%. - In almost all patients with normal placental blood flow extensive physiological vessel findings were recorded. In patients with reduced placental blood flow pathomorphological changes were found in 54%. - In diabetes of long duration with angiopathic complications - above all in White-classes D, F and R - the ratio of pathological findings in placental blood flow and morphology was increased. - But also in gestational diabetes and in disturbances of carbohydrate metabolism pathological results are to be found. Pathological results in placental blood flow were more often than morphological changes (70 versus 40%), so that also functional factors as uteroplacental blood flow influencing ones are to be taken into account.

The effect of intensified conventional insulin therapy before and during pregnancy on the malformation rate in offspring of diabetic mothers.

56 out of the 200 pregnant diabetic women admitted to our clinic between July 1981 and June 1983 had followed a pre-pregnancy metabolic intensive treatment programme. Most of these patients achieved near-normoglycemia: 87% or more of all their blood glucose readings before conception and in the early weeks of gestation were normoglycemic. The 56 patients were delivered of 57 babies, one of them suffering from fatal heart malformation. The 144 pregnant diabetics who were admitted to hospital only after eight weeks of pregnancy and had not had any special preconceptional metabolic control gave birth to 145 children, 9 of which presented congenital malformations: 3 of these were fatal another 3 were severe, and 3 were minor. These data are in line with our previously reported results on the years 1977-81. They stress the importance of a reasonably strict metabolic control, started well before conception, to prevent excess rates of congenital malformation.

The in vitro insulin secretion of human fetal pancreatic slices from diabetic and non-diabetic women--a methodical study.

The in vitro insulin secretion of pancreatic slices between the 11th and 15th week of pregnancy of fetuses from non diabetic ( FNDW ) and diabetic women ( FDW ) after incubation in media supplemented with different secretagogues was investigated in order to study the development of diabetic fetopathy during human pregnancy in diabetic women. There was a stimulatory effect on the insulin secretion in FNDW even if glucose alone was used, which became more pronounced if IBMX was added to the incubation medium. The insulin secretion was significantly enhanced in FDW compared to FNDW . This incubation model using fetal pancreatic slices seems to be appropriate for studying the ontogenesis of the human fetal pancreas.

[Effect, on the newborn infant, of carbohydrate metabolism in pregnancy in insulin-dependent diabetics]. / Der Einfluss des Kohlenhydratstoffwechsels während der Schwangerschaft insulinabhängiger Diabetikerinnen auf das Neugeborene.

The influence of the moment in pregnancy of insulin dependent diabetic women, at which normoglycemia by insulin therapy could be reached, on cord blood insulin concentration and neonatal morbidity was investigated. There is a positive correlation between the moment of normoglycemia (HbA1-values less than or equal to 8.5%) and the insulin concentration and furthermore the incidence of respiratory distress syndrome and macrosomia in the newborn. It is concluded that a tight metabolic control in insulin dependent diabetic women already prior to or early in pregnancy (at least until the 16th week) will be able to decrease the incidence of morbidity in infants of diabetic mothers.

Age-dependent insulin secretion of the endocrine pancreas in vitro from fetuses of diabetic and nondiabetic patients.

Fetal hyperinsulinemia is assumed to play a key role in the pathogenesis of diabetic fetopathy. To investigate the role of enhanced fetal B-cell mass as one cause of fetal hyperinsulinemia during diabetic pregnancy, we studied human fetal pancreatic slices from diabetic women (FDW) with poor metabolic control and nondiabetic women (FNDW) between 11 and 26 wk of pregnancy, morphometrically and by in vitro incubation experiments. Abortions had been performed due to different medical indications. We found a good correlation between the calculated B-cell mass and the gestational age in both FDW and FNDW, but the increase in FDW was much more pronounced. Such a correlation was also found in vitro regarding the insulin response to glucose and IBMX. The FDW had significantly higher values than FNDW of the same age range. In contrast to this, we found in two diabetic patients with tight metabolic control during the whole pregnancy results similar to those in FNDW. Therefore, we assume that it could be possible to prevent fetal hyperinsulinemia and perhaps even diabetic fetopathy in diabetic women by tight metabolic control during the whole pregnancy, but further investigations are necessary.

Prevention of congenital malformations in infants of insulin-dependent diabetic mothers.

From April 1977 to April 1981, 420 deliveries of infants of insulin-dependent diabetic women were performed in our department. Of the infants delivered, 23 had congenital malformations (5.5%). The malformation rate was 1.4% for infants of 420 nondiabetic women. Strict metabolic control was begun after 8 wk gestation in 292 of the diabetic women who delivered 22 infants with congenital malformations (7.5%). Intensive treatment was begun before conception in 128 diabetic women planning pregnancy. There was only one malformation in infants of this group (0.8%), a significant reduction from the anomaly rate in the late registrants (X2 = 7.84; P less than 0.01). These observations indicate that reasonable metabolic control started before conception and continued during the first weeks of pregnancy can prevent malformations in infants of diabetic mothers.

[Fetal hyperinsulinism in early pregnancy--a cause of diabetic fetopathy?]. / Der fetale Hyperinsulinismus in der Frühsehwangerschaft--eine Ursache der diabetischen Fetopathie?

The influence of fetal hyperinsulinemia on the development of diabetic fetopathy was investigated. Human fetal pancreatic slices of fetus between the 12th and 26th weeks of pregnancy from non diabetic (n = 32) and diabetic (n = 18) women were incubated in vitro for one hour. The insulin secretion results clearly demonstrate an age dependent increase in both groups investigated, but the increase is much more pronounced in fetuses from diabetic women. The differences between the groups are present already during the 12th week of pregnancy. The results support the concept to normalize the metabolic control in pregnancy as early as possible in order to prevent the diabetic fetopathy.

[Placental maturation disorders in relation to the quality of metabolic control during pregnancy in insulin-dependent and gestational diabetes]. / Reifungsstörungen der Plazenta in Relation zur Qualität der metabolischen Kontrolle während der Schwangerschaft beim insulinpflichtigen und Gestationsdiabetes.

In 1969/70 and 1979/80, 100 placentas each of diabetics were morphologically examined. The findings obtained were evaluated against the background of metabolic monitoring and typical phenomena of perinatal morbidity. The studies were conducted for the purpose of finding out the extent to which maintenance of normal glycaemic metabolism throughout pregnancy could affect the severity of diabetic disorders of placental maturation. Medium to severe placental maturation disorders were found to be closely correlated with quality of metabolic control (P less than 0.01). The findings thus obtained have produced evidence to the effect that maintenance of normal glycaemic metabolism throughout pregnancy would be of favourable impact not only upon perinatal mortality and morbidity. It would also cause significant reduction of placental maturation disorders.

[Studies into human foetal pancreas--critical appraisal of methods for tissue preparation (author's transl)]. / Untersuchungen am humanen fetalen Prankreas--Methodenkritik zur Gewebegewinnung.

Three different methods of legal abortion were applied to 23 patients, who had asked for termination of pregnancy, between the eleventh and 14th weeks of pregnancy. In vitro studies into human foetal pancreas were taken into consideration, when the choice was made. Carbohydrate tolerance was normal in all cases (50 g oGTT). Timing of intra-uterine foetal death, in the context of the three above methods (B-scan ultrasonic diagnosis and HPL level measurement) as well as studies into in vitro stimulation of foetopancreatic insulin secretion have shown purely mechanical, surgical abortion, sometimes following prostaglandin cervical gel priming, to be indispensable a condition for successful in vitro elucidation of the aetiopathogenesis of foetal diabetes. Extra-amniotic prostaglandin induction of abortion has failed to yield any relevant information, in that context.

PIP: 3 different methods of legal abortion were administered to 23 patients who had requested termination of pregnancy between weeks 11-14 of pregnancy. In vitro studies into human fetal pancreas were taken into consideration when the choice was made. Carbohydrate tolerance was normal in all cases (50 g oGTT). Timing of intrauterine fetal death, in the context of the 3 above methods (B-scan ultrasonic diagnosis and HPL level measurement), as well as studies into in vitro stimulation of fetopancreatic insulin secretion, have shown purely mechanical, surgical abortion, sometimes following prostaglandin (PG) cervical gel priming, to be indispensable as a condition for successful in vitro elucidation of the etiopathogenesis of fetal diabetes. Extraamniotic PG induction of abortion has failed to yield any relevant information in that context. (author's)

[Behaviour of human placental lactogen (HPL) following continuous glucose infusion in late pregnancy (author's transl)]. / Das Verhalten des HPL (Humanes plazentares Laktogen) unter Glukoseinfusionsbelastung in der Spätschwangerschaft.

Continuous intravenous glucose infusion was applied to 15 pregnant women of normal body weight, in the 37th week of pregnancy. The glucose levels in the blood, serum immunoreactive insulin, and placental lactogenic hormone (HPL) were monitored for three hours. Glucose tolerances were found to be normal in five probands, borderline in another five, and pathological in five, as well. No relationships were found to exist between insulin requirement und plasma HPL levels. While all HPL levels were somewhat increased in those patients with pathological glucose tolerances, the differences were not significant.