Antibodies against different epitopes of heat-shock protein 60 in children with type 1 diabetes mellitus.

The aim of this study was to investigate the amounts and epitope specificity of antibodies against heat shock protein 60 (hsp60) in the sera of type 1 diabetic and healthy children. Antibodies specific for peptide p277 of human hsp60 and of M. bovis as well as for human hsp60, M. bovis hsp65 proteins were measured by ELISA. Other autoantibodies (islet cell antibodies, glutamate decarboxylase antibodies and IA-2 antibodies) were also determined. A total number of 83 serum samples from children with type 1 diabetes mellitus and 81 samples of control children were investigated. Epitope scanning of the hsp60 for linear antibody epitopes was carried out using synthetic peptides attached to pins. The antibody levels specific for peptide p277 of human- and of M. bovis origin were significantly (human: P=0.0002, M. bovis: P=0.0044) higher in the diabetic children group than in the healthy children. We could not find significant difference in the antibody levels to whole, recombinant hsp proteins among the examined groups of children. Antibodies to two epitope regions on hsp60 (AA394-413 and AA435-454) were detected in high titres in sera of children with diabetes mellitus. The first region similar to the sequence found in glutamate decarboxylase, whereas the second one overlaps with p277 epitope to a large extent. Presence of antibodies to certain epitopes of hsp60 (AA394-413-glutamic acid decarboxylase-like epitope; AA435-454-p277-like epitope) in diabetic children may reflect their possible role in the autoimmune diabetogenic process of the early diabetes.

The pathophysiological influence of leptin and the tumor necrosis factor system on maternal insulin resistance: negative correlation with anthropometric parameters of neonates in gestational diabetes.

The contribution of the tumor necrosis factor (TNF) system and leptin was studied in insulin resistance and neonatal development during the course of normal pregnancy and gestational diabetes mellitus (GDM). Thirty patients with GDM and their neonates (n = 30), 35 healthy pregnant women (15 in the first, nine in the second and 11 in the third trimester) and their neonates (n = 20), and 25 healthy matched non-pregnant women participated in the study. Significantly elevated levels of maternal TNF-alpha, sTNF receptor (R)-1 and R-2, leptin (detected by enzyme-linked immunosorbent assay) and fasting C-peptide (measured by radioimmunossay and raised body mass index (BMI) were found in GDM patients and in the third trimester of normal pregnancies. TNF-alpha, sTNFR-2, C-peptide, leptin concentrations and BMI positively correlated with each other in GDM. An inverse relationship between the body length, head circumference and body weight of the newborns, and maternal TNF-alpha, leptin and C-peptide concentrations was shown in GDM. In healthy pregnancies the maternal serum leptin level was in a negative linear correlation with the head circumference of the newborns. In conclusion, increased TNF-alpha and leptin levels may contribute to insulin resistance in GDM and in the third trimester of normal pregnancy and may negatively influence the anthropometric parameters of the newborns.

[Detection of antibodies against pancreatic islet cells in clinical practice]. / Hasnyálmirigy-szigetsejt elleni antitestek vizsgálata a klinikai gyakorlatban.

Development of diabetes mellitus caused by pancreatic beta-cell destruction of autoimmune origin is the result of a long lasting process. The most easily examinable feature of this stage is the occurrence of the islet cell antibodies. The sera which are positive for islet cell cytoplasmic antibodies (ICA), examined by indirect immunofluorescence, contain a mixture of antibodies. The glutamic acid decarbocylase (GAD), the tyrosin phosphatase (IA2), the insulin, and the GM2-1 glycolipid can be the targets of these antibodies. One can routinely examine the ICA, the GADA, the IA2 antibodies. The detection of antibodies against insulin (IAA) and GM-2-1 glycolipid is not invented in the routine laboratory work. The aim of the authors was the evaluation of clinical significance of occurrence of islet cell antibodies: one hundred and eighteen nondiabetic children an adult human being without known diabetic first degree relatives and 366 type 1 diabetic children and adult patients served as controls. The authors evaluated the predictive value of the different islet cell antibodies to the development of type 1 diabetes mellitus in 596 nondiabetic children with type 1 diabetic first degree relatives. The authors looked for markers of beta-cell destruction among sera of 320 diabetics manifested after 30 years of age with at least half a year of non-insulin-dependency and in the sera of 68 females suffered from gestational diabetes after 0-14 years of the index pregnancy. Finally the authors report 7 cases in which the examination of islet cell antibodies helped the diagnosis and classification of diabetes mellitus. Indirect immunofluorescence method was used for the detection of ICA, radioimmunoassay for that of GADA and IA2 antibodies. There was no positive reaction for ICA and GADA in the nondiabetic population without diabetic first degree relatives. Among the freshly diagnosed type 1 diabetic children 39% were positive for only ICA, 44% for only GADA and 80% for any antibodies. Among the freshly manifested type 1 diabetic adults ICA positivity only was observed in 21%, GADA positivity only in 7.1% and 93% for any antibodies. From the 595 nondiabetic children with type 1 diabetic first degree relatives 23 were positive for ICA, from whom 5 became diabetic during a two years observation period. These diabetic children had multiplex autoantibodies besides ICA. One child from this group, who was negative for ICA became diabetic, too. Among type 2 diabetic patients 13% were positive for ICA alone, 17% were positive for GADA alone and 27% were positive for any antibodies. The insulin dependency manifested in a short time was associated with antibody positivity. Among the gestational diabetics 10 were found positive for ICA. From them, 7 were type 1 diabetics, and 3 were type 2 diabetics at the time of the detection of antibodies. The authors suggest the need of determination of islet cell antibodies in the group of nondiabetic first degree relatives of type 1 diabetic patients (ICA, GADA, IA2 and IAA), in the group of non-insulin-dependent diabetics (ICA and GADA) as a screening for later insulin dependency, and in gestational diabetes after delivery (ICA) as screening for type 1 diabetes mellitus.

[Ectopic ureter in infancy and childhood]. / Ureter ectopia csecsemökben és gyermekekben.

Two infants, 7 children and 1 young woman have been surgically treated for ureteral ectopy in the Departments of Paediatrics and of Urology of the Medical University of Pécs over the last 15 years (1974-1989). The girl/boy ratio was 8/2. In girls, who were otherwise toilet-trained with a normal voiding pattern, constant wetting and urinary infection were the leading clinical findings. The site of ureteral drainage was the vestibule in 4 patients, the urethra in 3, the vagina in 1, the prostatic utricle in 1, and it could not be determined in 1 girl. Diagnosis was based on intravenous urography, voiding cystourethrography, ultrasonography, isotope scan, endoscopy and filling up of the bladder with a methylene blue solution. The diagnosis was more obscure when the ectopic ureter drained a poorly functioning kidney. Considering that in ectopy with duplicated system the upper pole renal segment is almost always destroyed, upper pole nephrectomy and proximal ureterectomy are advocated. In 1 neonate with esophageal atresia and tracheo-esophageal fistula ultrasonography detected the ureteral malformation. In 1 girl bilateral single ureteral ectopy was found.

Early prediction of fetal macrosomia in diabetes mellitus.

We studied 374 pregnant diabetic women to determine the value of various ultrasound parameters in the prediction of fetal macrosomia. The correlation between ultrasonographic signs and maternal glycaemia in the development of fetal macrosomia was also studied. Significant correlation was observed between the accurence of hydramnios and future macrosomia during the second-trimester (p less than 0.001). Serum fructosamine levels as an index of maternal glycaemia in patients of macrosomic fetuses were significantly higher throughout the pregnancy as compared with mothers of infants with normal birth weight (p less than 0.001). These data suggest: 1. The presence of hydramnios in the second trimester is a useful predictor of macrosomia in diabetic patients (specificity: 86%, negative predictive value: 88%). 2. Maternal diabetic control during pregnancy has a significant influence on fetal growth and contributes to the development of fetal macrosomia. 3. The lack of correlation between the frequency of hydramnios and fructosamine levels suggests that a mechanism other than carbohydrate metabolism also plays an important role in the development of fetal macrosomia.

Studies of dental and oral changes of pregnant diabetic women.

The longitudinal examination of 132 pregnant diabetic women under care showed a 96.2% prevalence of gingivitis. The intensity of gingivitis was most marked in weeks 11 to 15, and 24 to 26 of pregnancy, and the correlation with changes in oral hygiene was statistically significant (p less than 0.001). On the other hand, the severity of diabetes had no effect on the degree of gingival inflammation. As for caries, the mean DMF values increased during diabetic pregnancy, the number of carious (D) and filled (F) teeth to a higher, that of extracted (M) teeth to a lesser degree, than in diabetic non-pregnant women.

Investigation on serum C-peptide concentrations in pregnant diabetic women and in newborns of diabetic mothers.

Serum C-peptide concentrations at delivery and neonatal complications were investigated in a group of diabetic mothers and in their infants (IDM, n = 29). Furthermore, the changes in B-cell function and in daily insulin demand was followed up in 12 pregnant diabetics (of them 8 with long-term and 4 with short-term and "mild" diabetes) during pregnancy. All these diabetic mothers were under an intensive metabolic control with the aim to achieve normoglycaemia mainly in the second part of pregnancy. Mean cord blood C-peptide level of 29 IDM was 0,58 +/- 0,43 nmol/l, being not significantly higher than either the average C-peptide concentration of our adult control group (0,50 +/- 0,15 nmol/l, n = 24) or that of seven infants born to healthy mothers (0,58 +/- 0,37 nmol/l). Early hypoglycaemia was observed in five, macrosomia in three and IRDS in one neonate, resp. mothers coming to our intensive care after the 13th week of gestation gave birth with a higher incidence of neonatal complications than those controlled already in the first trimester or even preconceptionally. In 20 of 25 mothers studied venous C-peptide concentrations at delivery were undetectably low, in spite of their infants having cord blood C-peptide levels in the measurable range. In 8 of the 12 diabetic mothers with long-term diabetes C-peptide levels remained under the detection limit of the assay throughout pregnancy. In the 4 other cases with "mild" diabetes (and, hence, with a late start of intensive control) C-peptide values increased in the course of pregnancy; however, 3 of the four mothers gave birth with neonatal complications. These results indicate that (1) early--preferably preconceptional--intensive metabolic control of pregnant diabetics may reduce the incidence of neonatal complications; (2) fetal C-peptide can neither during pregnancy nor at birth pass through the placental barrier, and (3) mothers with "mild" diabetes require the same early and strict metabolic control as the more severe cases to avoid neonatal complications.

Care of pregnant diabetics: medical aspects.

A method has been worked out for the intensive care of pregnant diabetics with the object of preventing damage both to the mother and the fetus. The method requires close cooperation between a team of doctors including obstetrician, internist and pediatrician. The present paper reports the experience of the first two years of application of the method and refers to a total of 75 women. The main point consists in an attempt to maintain a normal blood glucose level by administering increasing doses of insulin. Periods of inpatient management alternated with close outpatient control. Of the 75 insulin-dependent diabetics 45% had severe diabetes (White classes D-F); nevertheless, perinatal mortality was only 5.78 per cent. It is worth stressing that the 25 diabetics who came under intensive care before conception or in the early stages of pregnancy all gave birth to live healthy babies.

Potentiation of glibenclamide-induced insulin release by calcium infusion.

The effect of calcium on glibenclamide-induced insulin release was studied in 14 diabetic patients. Two mg glibenclamide was given intravenously and calcium, blood glucose and IRI were determined in venous blood samples at predetermined intervals. The test was repeated 3-4 days later with the patients simultaneously receiving a calcium infusion into a contralateral vein. The decrease in blood glucose and the rise in IRI level were both significantly greater in the combined glibenclamide-calcium test. It is concluded that calcium may temporarily improve carbohydrate tolerance in diabetic patients by potentiating the glibenclamide-stimulated insulin secretion.

Study on the insulin-binding capacity of blood serum by a cellulose adsorption method.

The cellulose adsorption method was used for the quantitative study of the insulin-binding capacity of blood serum. The results support the view that anti-bodies to insulin are of exogenous origin and that may be responsible for insulin resistance in individual cases. While the insulin requirement of patients was found to be related to the serum antibody level, the latter was unaffected by the duration of insulin treatment even though the number of cases exhibiting normal values decreased with the length of treatment.