Detection of proinsulin, C-peptide, insulin-A-chain, and glicentin in pancreatic islet cells of early human fetogenesis.

The presence of C-peptide, proinsulin, insulin-A-chain, and glicentin in human fetal pancreatic cells by using the PAP-technique was investigated and the results obtained compared with the occurrence of insulin or glucagon immunoreactive cells. In pancreatic sections obtained from 10 weeks old human fetuses we could identify cells reacting with antibodies directed against C-peptide, proinsulin, and insulin-A-chain. The majority of the cells were found in the duct epithelium and their number increased from the 10th to 14th week forming clusters near the ducts. The number and localization of the cells correspond exactly to the insulin positive cells. The presence of proinsulin and insulin-A-chains is a further proof of biological activity already in an early step of fetal development. The presence of glicentin-positive cells in the 10th week of gestational age as well as cells reacting with glucagon antibodies provide evidence for active glucagon biosynthesis. The number of these cells increased markedly in the 14th week of gestational age.

[A case of septic ovarian vein thrombosis]. / Ein Fall von septischer Ovarialvenenthrombose.

Puerperal ovarian vein thrombosis commonly originates from purulent necrotic endomyometritis. The incidence is published to be 1 to 600 deliveries. According to the puerperal uterine drainage, the predominant location is the right ovarian vein in 90% of all cases. The leading symptoms are lower abdominal pain, fever and leucocytosis. Discrepancy between the given clinical picture and the insignificant findings on gynaecologic examinations is common.

Human amniotic fluid stimulates DNA synthesis of rat pancreatic islets.

Collagenase isolated pancreatic islets from newborn Lewis rats were used to study the effect of human amniotic fluid (HAF) on DNA synthesis measured as incorporation of 3H-thymidine into islet DNA and as estimation of labeling index. We investigated individual samples of HAF obtained from pregnant women between the 28th and 39th week of gestation. Since there were no significant differences of HAF obtained from different gestational weeks on islet DNA synthesis, the following experiments were carried out with pooled HAF. When islets were cultured for 2 days in TCM 199 containing 10 mmol/l glucose the addition of 50% HAF resulted in a significantly increased DNA synthesis in comparison to islets cultured in TCM 199 supplemented to 50% with Hank's balanced salt solution (HANK). When extending the culture period from 2 to 4 days the stimulatory effect of 50% HAF was well preserved. Islets cultured in HANK are characterized by a declined DNA-content, which was prevented in the presence of HAF. There was a good correlation between incorporation of 3H-thymidine and the percentage of labeled nuclei, estimated on hematoxylin/eosin-stained autoradiograms of islets cultured for 4 days either in TCM 199 alone, in the presence of 50% HANK or 50% HAF, indicating that the measured incorporation of labeled thymidine into islet DNA was representative for islet replication.

Immunohistochemical, morphometric, and ultrastructural investigations of the early development of insulin, somatostatin, glucagon, and PP cells in foetal human pancreas.

Fresh autopsy specimens of pancreas, taken from 18 human foetuses at the 10th (n = 4), 12th (n = 7), and 14th (n = 7) weeks of gestation, were analyzed immunohistochemically for the presence of islet parenchymal cells, immunoreactive with antisera raised against insulin (B cells), somatostatin (D cells), glucagon (A cells), and pancreatic polypeptide (PP cells). All four islet cell types were found sporadically within or near the epithelium of the small excretory ducts at the 10th week of development. At the 12th week, their presence appeared to be no longer restricted to the duct epithelium, as some B cells were found also in small clusters outside the ducts. At the 14th week of development, the B cells formed large clusters in the neighborhood of the excretory ducts. It is at this stage that the first parenchymal cells with Grimelius argyrophilia could be found. They were supposed to represent A cells. The B cells were found to be the predominating type of islet cells (about 50%) at the 10th week of gestation. The relative volume density was about 25% for the D cells, about 15% for the A cells, and about 10% for the PP cells. At the 12th and 14th weeks of development, the relative numbers of B and PP cells decreased somewhat (to 36 and 6%, respectively), whereas those of the D and A cells were found to increase (to 30 and 27%, respectively). The relative volume density of the total islet parenchyma was about 2, 6, and 21% at the 10th, 12th, and 14th weeks of development, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of human amniotic fluid on DNA synthesis of rat pancreatic islets in tissue culture.

In this study we investigated the effect of human amniotic fluid (HAF) on the incorporation of 3H-thymidine into cultured rat islets. The addition of 1% HAF (1% HAF, 99% TCM) did not alter the incorporation of labeled thymidine, in comparison to TCM 199 alone (100%). The culture in the presence of 50% HAF (1:1 diluted with TCM, v/v) resulted in a significantly increased DNA synthesis. To exclude any effect of ionic composition or substrate dilution by the HAF, the TCM 199 was also either diluted (1:1) with saline or Hank's balanced salt solution (HBSS). Islets cultured under these conditions did not show any stimulated thymidine incorporation into islet DNA. From these results we conclude that HAF contains an activity which stimulates islet replication. In order to concentrate this stimulatory activity, HAF was tested after it had been lyophilized or evaporated. Independent of the method used, in no case a stimulatory effect of concentrated HAF was found. In contrast, the concentrated form of HAF inhibited the incorporation of labeled thymidine into islet DNA.

Neuron-specific enolase (NSE) as a neuroendocrine cell marker in the human fetal pancreas.

Using the PAP technique, we investigated the presence of neuron-specific enolase in the human fetal pancreas of 10, 12, and 14 weeks of gestational age. Neuron-specific enolase is present in the islet cells in the 10th week. Positive cells are situated mainly in duct epithelium. The number of cells with a positive reaction increases from the 12th to the 14th week. In the 14th week, they are clustered either near the ducts or between the acini. The numbers and localizations of the cells correspond to those obtained in previous studies with 4 basic islet cell types in the same material. The present results are a further proof that islet cells are biologically active during early fetal development.

[Immunocytochemical studies of the development of basal cell types of human islet organs]. / Immunzytochemische Untersuchungen zur Entwicklung der Grundzelltypen des menschlichen Inselorgans.

The investigations are carried out in 19 human fetal pancreases. The detection of the 4 islet hormones insulin, glucagon, somatostatin and PP ist carried out in PAP-technique. The parts of these 4 types of islet cells are estimated quantitatively. In the 10th to 15th week of development insulin-producing B-cells are present. Moreover glucagon-, somatostatin- and PP-cells in the islet organ are present. In the group of 16th to 20th week of gestation insulin-, glucagon- and somatostatin-cells are increased compared to the first group. PP-cells are not altered. The increase of 3 types of islet cells is a result of fetal development.

Phases in the early development of the human islet organ.

The study included morphometric examination and biochemical investigation of 41 human pancreata taken from 14th to 26th weeks of fetal development. 3 phases of development were observed. Phase I (weeks 14 to 16) is characterized by the presence of mainly islet buds. They were originated from the ducts and are vascularized during week 16. During phase II (17th to 20th weeks) the islet buds were detached from the ducts and they formed new mantled islets with the B cells in the centre. Non-B cells are situated on the periphery around the insulin producing cells. Phase III lasts from week 21 to week 26. During this phase B cells and non-B cells become more irregularly positioned within the islet, a cytology, which is similar approximating that of as also found in adult human islets. The changes of islet structure during pancreatic development are accompanied by other typical phenomena: Islet size increases during phases I and II, but decreases again during phase III. The proportion of isolated B cells outside of the islets varies during this stage of fetal development, but they generally account for about 15% of the total islet organ. This should be taken into account when assessing the islet function De Pablo et al. (1985) on a morphological basis.

Tissue cultivation as a method for preservation of human foetal islet parenchyma--a correlated biochemical, immunohistochemical and morphometric investigation.

Tissue culture of human foetal pancreas slices, obtained at a gestational age between 10 and 19 weeks, as a method of preservation and to pool the material before transplantation was investigated. Before and after 2 weeks of culture the pancreatic insulin content, the insulin secretion in response to glucose and isobutylmethylxanthin (IBMX) as well as the protein biosynthesis were measured. In addition, the distribution of the insulin immunoreactive cells was examined, as well as their relative volume density. After 2 weeks' culture an increase of the basal insulin secretion was observed; this was probably due to the glucose content (10 mmol/l) in the culture medium. Neither the stimulated insulin secretion, nor the protein biosynthesis, the insulin content, and the B-cell volume were altered by the used culture conditions. It was concluded that tissue culture is a suitable method to preserve human foetal pancreas slices before transplantation.

[In vitro studies of amniotic fluid and intrauterine growth retardation]. / In vitro Untersuchungen von Fruchtwasser und intrauterine Retardierung.

Until now the intrauterine growth retardation remains as an insufficiently solved problem. The 3H-thymidine incorporation into the DNA of the isolated islets of Langerhans of neonatal Lewis rats was used as a marker for cell dividing activity in vitro. There was no inhibition effect on the 3H-thymidine incorporation in the presence of amniotic fluid from pregnancies with intrauterine growth retardation compared to those of normal pregnancies.

[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.

Further evidence for a preventive therapy of insulin-dependent diabetes mellitus in the offspring by avoiding maternal hyperglycaemia during pregnancy.

A highly significantly decreased prevalence of insulin-dependent childhood-onset diabetes (less than 1/3 of the initial prevalence rate) could be achieved in Berlin/GDR since 1973 by improving systematically diagnostic and therapeutic measures for pregnant diabetics, particularly for non-insulin-dependent gestational diabetics. In addition, a highly significantly increased incidence rate of diagnosed, diet-treated and delivered non-insulin-dependent pregnant diabetics was found between 1979 and 1983 in Berlin/GDR, Halle and Leipzig as compared to the other districts of the GDR. Simultaneously, a highly significantly decreased prevalence rate of diabetic children (less than 1/3), who were born during this period, was found in 1983 for Berlin, Halle and Leipzig as compared to the other districts of the GDR. Finally, a highly significant inverse correlation could be demonstrated for the 15 districts of the GDR between the incidence rates of diagnosed, diet-treated and delivered non-insulin-dependent pregnant diabetics and the prevalence rates of diabetic children who were born during this period (1979-1983). In view of these findings, an interruption and even a reversal of the continued dramatic increase of idiopathic insulin-dependent diabetes mellitus appears to be possible in the developed countries by preventing maternal hyperglycaemia during pregnancy and hence hyperinsulinism in the foetuses and newborns.

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.

[Quantitative-histologic studies of human fetal pancreas from metabolically healthy and insulin-dependent diabetic women]. / Quantitativ-histologische Untersuchungen des fetalen menschlichen Pankreas von stoffwechselgesunden Frauen und insulinabhängigen Diabetikerinnen.

Human fetal pancreases from nondiabetic (n = 26) and insulin-dependent diabetic women (n = 14) between the 13th and 26th week of pregnancy were investigated morphologically. There was in enhanced islet cell volume in fetuses from diabetic patients compared to those from nondiabetics, mainly caused by nesidioblastosis. This phenomenon could be caused by the metabolic state of the insulin-dependent diabetic patients. There were also single beta cells or groups of beta cells, which could be found between the exocrine pancreatic tissue, already at an early stage in pregnancy. It is assumed, that the nesidioblastosis in fetuses from diabetic women is caused by a continuous stimulation of the fetal endocrine pancreas during pregnancy. These results are in a good correlation to the in vitro results of the insulin secretion of human fetal pancreatic slices after incubation.