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OBJECTIVE Insulin resistance might play a role in the pathogenesis of polycystic ovarian syndrome (PCOS). The family of glycoprotein 130 (gp130) cytokines could influence insulin action. One of these cytokines is interleukin (IL)-6, which exerts a short-term insulin-sensitizing effect, whereas in a long-term period, it might induce insulin resistance. Some other gp130 activators are supposed to have beneficial metabolic effects. Gp130 is present in the circulation in the soluble form (sgp130), which inhibits intracellular gp130 signaling. The aim of the present study was to estimate the relation between sgp130 and insulin sensitivity in women with PCOS. RESEARCH DESIGN AND METHODS We studied 78 women with PCOS (35 lean and 43 obese) and 34 healthy women (18 lean and 16 obese). The euglycemic-hyperinsulinemic clamp and the measurements of serum sgp130, IL-6, soluble IL-6 receptor (sIL-6R), and sex hormones were performed. RESULTS Both obesity and PCOS were characterized by an increased sgp130 (P < 0.0001 and P = 0.0002, respectively). sIL-6R concentration was lower (P = 0.0036) in women with PCOS independently of obesity. Serum sgp130 was negatively correlated with insulin sensitivity when control and PCOS women were analyzed together (r = −0.36, P < 0.0001) and in the PCOS subjects separately (r = −0.34, P = 0.002). In multiple regression analysis, this correlation was significant after adjustment for BMI, waist, percent of body fat, postload glucose and insulin, triglycerides, and high-sensitive C-reactive protein. CONCLUSIONS Serum sgp130 is inversely and independently associated with insulin sensitivity in women with PCOS. An increased serum sgp130 in obesity and PCOS suggests an inhibition of intracellular gp130 signaling in insulin-resistant conditions.

Background Diabetes mellitus affects the metabolism of several components of extra-cellular matrix, including glycosaminoglycans (GAG). Alterations in the metabolism of GAG may play an important role in the development of diabetic complications. Methods Consequently, the relationship between diabetic nephropathy and urinary GAG excretion has been estimated in 86 diabetic patients (33 type 1 diabetes mellitus (DM), 53 type 2 DM) in comparison to 30 healthy controls (Figure 1). GAG concentration in 24-h urine samples has been determined by precipitation with cetylpyridinum chloride and potassium acetate in ethanol followed by a fluorometric test with 2-Hexadecyl-9H-pyrido(4,3b)indolium Bromide. Results Diabetic subjects excrete significantly more GAG than the control group (66.47 mg/24 h vs 50.11 mg/24 h). A marked difference in urinary GAG excretion between diabetic patients with nephropathy (74.66 ± 7.5 mg/24 h) and without nephropathy (50.13 ± 5.37 mg/24 h) has been detected. With diabetic nephropathy, patients with a longer history of GAG excretion experience an increase. An increased urinary GAG excretion has been detected in patients with microalbuminuria or macroalbuminuria. Conclusions In conclusion, it can be stated that all patients with DM compared to the control group show an increase in GAG excretion independent of diabetes duration. Urinary GAG excretion positively correlates with the duration of diabetic nephropathy. The assessment of GAG excretion values may be useful for determining the early stages of diabetic nephropathy and the progression of the disease. Copyright © 2008 John Wiley & Sons, Ltd.

Objective: There is growing evidence that adiponectin function is related to the pathogenesis of insulin resistance. Insulin resistance might be present even in lean subjects with a strong family history of type 2 diabetes. The aim of the study was to look for adiponectin’s role in the pathogenesis of insulin resistance in offspring of type 2 diabetic patients, and its relation to the activity of the tumor necrosis factor (TNF)-α system. Research design and methods: The study was carried out in 23 lean offspring of type 2 diabetic subjects and in 23 controls matched for age, sex and body mass index. The oral glucose tolerance test for glucose and insulin estimations and hyperinsulinemic, euglycemic clamp studies were performed in all patients. The plasma concentration of adiponectin, TNF-α, soluble TNF receptors 1 and 2 (sTNFR1, sTNFR2), HbA1c, total cholesterol, high-density lipoprotein (HDL)-cholesterol, low-density lipoprotein-cholesterol and triglycerides were estimated. Results: The insulin sensitivity index, normalized for fat-free mass (Mffm) and adiponectin concentrations were markedly decreased in offspring of type 2 diabetic subjects compared with the control group (P = 0.0046 and P = 0.00 058 respectively). TNF-α and sTNFR1 concentrations did not differ between the studied groups; however the concentration of sTNFR2 was markedly increased in the offspring of type 2 diabetic patients (P = 0.0002). Adiponectin concentration was positively correlated to the insulin sensitivity index (r = 0.34; P = 0.020) and to HDL-cholesterol (r = 0.29, P = 0.047) and was inversely related to sTNFR2 (r = −0.33, P = 0.027). Conclusions: The obtained results suggest that adiponectin could play a role in the pathogenesis of insulin resistance in lean offspring of type 2 diabetic subjects.

Plasma concentrations of adiponectin, tumor necrosis factor-alpha (TNF-alpha) and its soluble receptors sTNFR-1 and sTNFR-2 were measured in 80 patients with gestational diabetes (GDM) (mean age 29.0 +/- 4.9 years) and 30 pregnant women with normal glucose tolerance (NGT) (mean age 28.2 +/- 6.0 years). We found that GDM patients had significantly lower concentrations of adiponectin (11.28 +/- 5.91 vs. 16.31 +/- 6.04 microg/ml, p = 0.00009) and elevated levels of TNF-alpha (1.71 +/- 0.92 vs. 1.27 +/- 0.42 pg/ml, p = 0.0175) in comparison to NGT women. The differences remained statistically significant after adjusting for BMI. Plasma levels of sTNFR-1 and sTNFR-2 also tended to be higher in GDM patients. In the GDM group TNF-alpha concentrations correlated significantly with sTNFR-1 (r = 0.444, p = 0.00008), sTNFR-2 (r = 0.364, p = 0.0016) and with C-peptide concentrations (r = 0.318, p = 0.016), whereas in women with NGT TNF-alpha correlated only with TG levels (r = 0.50, p = 0.024). Multivariate linear regression analysis revealed that prepregnant BMI was the most predictive indicator of TNF-alpha concentrations in GDM women. TG concentrations as well as BMI before pregnancy and at the time of sampling in pregnant NGT women were significant predictors, explaining 62% of the variance in TNF-alpha concentration. There were also negative correlations between adiponectin concentrations and a pregestational BMI (r = - 0.298, p = 0.009), BMI at the time of sampling (r = - 0.239, p = 0.034) and TG concentrations (r = - 0.379, p = 0.039) in GDM patients, whereas women with NGT showed only a negative correlation between adiponectin and TG concentrations (r = - 0.488, p = 0.025). In a multivariate regression analysis, prepregnancy BMI and TG levels remained significant predictors, explaining 39% of the variation in plasma adiponectin concentration in GDM women. In conclusion, our results suggest that decreased adiponectin concentration in GDM may not simply reflect maternal adiposity and insulin resistant state, but may contribute to the impaired glucose metabolism during pregnancy, with potential implications for screening and prevention of the disease.

Summary Objective Hyperhomocysteinemia is a well known risk factor for the diseases of the cardiovascular system, which seem to be the main cause of increased mortality in patients with type 2 diabetes. The aim of the study was to evaluate the levels of homocysteine in patients with type 2 diabetes in respect to the regimen of diabetes treatment as well as the presence of diabetic complications. Methods The investigation was carried out in the group of 64 patients with type 2 diabetes and in 18 healthy subjects from the control group. Clinical examination and measurements of homocysteine, folic acid, vitamin B12, glycosylated hemoglobin concentration and evaluation of parameters of the lipid metabolism, microalbuminuria and creatinine were done in both groups. Results Homocysteine concentration was significantly higher in the group of patients with diabetes in comparison to the control group (p = 0.0007). Diabetic patients had significantly lower concentrations of folic acid (p = 0.028) and HDL cholesterol (p = 0.025) together with higher levels of systolic blood pressure (p = 0.007). In the group of patients with diabetes no differences in homocysteine levels were found in respect to diabetes treatment. Diabetic patients with coronary artery disease had significantly higher homocysteine concentration in comparison to the group with diabetes without history of coronary artery disease (p = 0.0097). Homocysteine levels correlated significantly with incidence of ischaemic heart disease (r = 0.44, p = 0.001) and microalbuminuria (r = 0.26, p = 0.019). Negative correlation was noticed in HDL concentrations (r = -0.30, p = 0.013) and the levels of folic acid (r = -0.30, p = 0.008). Conclusion Our results suggest that hyperhomocysteinemia in diabetic patients may contribute to the development of chronic complications. The influence of diabetes treatment on Hcy levels requires further observations.

In vitro studies revealed that insulin resistance might be associated with the intracellular formation of ceramide, the second messenger in the sphingomyelin signaling pathway. The aim of the present study was to examine the content and composition of fatty acids in ceramide and sphingomyelin in human muscle and to evaluate their relationships with insulin sensitivity. The study was conducted on 27 male subjects with normal glucose tolerance. Euglycemic-hyperinsulinemic clamps and biopsies of vastus lateralis muscle were performed. In 10 subjects, additional biopsies were taken after a 4-h clamp and after a clamp with concurrent Intralipid/heparin infusion. We identified 13 ceramides and sphingomyelins according to fatty acid residues. Insulin sensitivity was related to total ceramide content (r = −0.49, P = 0.01) and to ceramide consisting of palmitic (r = −0.48, P = 0.011), palmitoleic (r = −0.45, P = 0.019), mirystic (r = −0.42, P = 0.028), and nervonic acid (r = −0.39, P = 0.047). Hyperinsulinemia did not affect estimated muscle parameters. Intralipid/heparin infusion resulted in a 24.73% decrease in insulin sensitivity (P = 0.007) and a 47.81% increase in ceramide content (P = 0.005). These changes were significantly related to each other (r = −0.64, P = 0.046). A relationship with the decrease in insulin sensitivity was also observed for ceramides consisting of palmitic (r = −0.68, P = 0.03) and linoleic (r = −0.66, P = 0.038) acid. Our data indicate that the sphingomyelin signaling pathway in muscle might be an important factor determining the development of insulin resistance in humans.

TNFalpha was shown to play an important role in the autoimmune inflammatory process of Graves' ophthalmopathy (GO). In our previous study we found no significant changes in serum TNFalpha levels in GO patients. The aim of the present study was to estimate an influence of corticosteroids on serum levels of TNFalpha receptors (sTNFR1 and sTNFR2) in GO patients and to assess their potential as a guideline of immunosuppressive therapy. We detected serum sTNFRI and sTNFR2 in three groups of subjects: 18 patients with clinical symptoms of ophthalmopathy [Clinical Activity Score (CAS)or = 4, anamnesis of GOor = 1 yr], 16 patients with Graves' disease without ophthalmopathy (Gd) and 14 healthy volunteers. Corticosteroid therapy consisted of intravenous infusions of methylprednisolone (MP) and subsequent treatment with oral prednisone (P). The serum samples were collected 24 hours before MP, 24 hours after MP, 14 days of treatment with prednisone and after the end of the corticosteroid therapy. The levels of serum sTNFR1 and sTNFR2 were determined by ELISA. Serum levels of sTNFR1 were significantly higher in GO individuals as compared to the control group (p0.01). We have found a significant decrease in sTNFR1 concentration in corticosteroid-respondent patients (satisfactory clinical effect, decrease of CASor = 1) as compared to the pretreatment values after MP treatment (p0.05) and after 14 days of prednisone (p0.01). There were significant differences in sTNFR2 level after MP treatment (p0.02) and after corticosteroid administration (p0.05) between responders and non-responders. Baseline values of sTNFRI in GO individuals were positively correlated with CAS (r = 0.6, p0.02).TNFalpha acting through its receptors plays an important role in the pathogenesis of Graves' ophthalmopathy. Moreover, the beneficial influence of corticosteroids on the course of GO may be explained, at least in part, by an inhibition of sTNFR1 and sTNFR2. Measurement of soluble TNFalpha receptors might potentially serve as an indicator in prognostic estimation of corticosteroids' efficacy.

OBJECTIVE— Tumor necrosis factor-α (TNF-α) is a possible link between obesity and impaired glucose tolerance (IGT) and type 2 diabetes. Data about TNF-α and soluble forms of its receptors (sTNFR1 and sTNFR2) in IGT are controversial. The aim of the present study was to assess plasma TNF-α, sTNFR1, and sTNFR2 levels and to evaluate the relationships with insulin resistance in obese subjects with IGT. RESEARCH DESIGN AND METHODS— A total of 104 subjects participated in the present study: 30 obese subjects with IGT (obese-IGT), 32 obese subjects with normal glucose tolerance (obese-NGT), and 42 lean healthy control subjects (control-NGT). Anthropometry and blood biochemical parameters were measured and euglycemic-hyperinsulinemic clamp was performed. RESULTS— Obese-IGT subjects were more insulin resistant in comparison with obese-NGT and control-NGT groups; obese-NGT subjects were more insulin resistant than control-NGT. Plasma sTNFR1 and sTNFR2 were markedly higher in both groups of obese subjects in comparison with control-NGT and in the obese-IGT versus obese-NGT group. Plasma sTNFR1 and sTNFR2 were inversely related to insulin sensitivity. Both relationships remained significant after adjustment for age, BMI, waist girth, percent body fat, plasma glucose, insulin, nonesterified fatty acids, cholesterol, and triglycerides. Correlation between sTNFR2 and insulin sensitivity was also present in all the groups analyzed separately, but the correlation between sTNFR1 and insulin sensitivity was present only in the obese-NGT group. CONCLUSIONS— Our data suggest that TNF-α receptors are increased in obese-IGT subjects and are related to insulin resistance. These findings indicate that the TNF-α system might contribute to the development of insulin resistance in glucose-intolerant subjects.

Obesity is associated with the increased risk of cardiovascular disease; however, mechanisms responsible for such an increase are not fully understood. IL-8 is a cytokine that might have atherogenic properties. Recent in vitro studies revealed that IL-8 is produced and secreted by human adipocytes. The aim of the present study was to evaluate plasma IL-8 concentrations in obese subjects and the relationships between circulating IL-8 and anthropometric and biochemical parameters and TNF-alpha system. A total of 75 subjects with normal glucose tolerance, 35 lean and 40 obese, were recruited for this study. Plasma IL-8 levels were measured in fasting state, after an oral glucose tolerance test and after the euglycemic hyperinsulinemic clamp. A significant increase in plasma IL-8 was observed in the obese group. In simple regression analysis, performed for the initial evaluation of relationships, plasma IL-8 was related to body mass index, percentage of body fat, fat mass (FM), and soluble TNF-alpha receptor 2 (sTNFR2) in both groups and with waist-to-hip ratio and sTNFR1 in the obese. In multiple regression analysis, FM, waist-to-hip ratio, gender, sTNFR2, and low density lipoprotein cholesterol were responsible for 44% of IL-8 variability. During oral glucose tolerance testing, mean plasma IL-8 concentrations increased in both groups, whereas clamp resulted in a significant increase in plasma IL-8 only in the obese. We conclude that plasma IL-8 levels are increased in obese subjects, and are related to FM and TNF-alpha system. Increase in circulating IL-8 might be one of the factors linking obesity with greater cardiovascular risk.

Circulating forms of l -selectin were found to be elevated in several autoimmune diseases. ICAM-1 has been suggested a predictor of the onset of GO. The aim of the study was an estimation of serum l -selectin and ICAM-1 in patients with Graves’ disease with ophthalmopathy during treatment with corticosteroids to assess their potential as a guideline of immunosuppressive therapy. We detected serum l -selectin and ICAM-1 in three groups of subjects: 20 patients with Graves’ disease without ophtamopathy (Gd), 17 patients with clinical symptoms of ophthalmopathy (CAS≥3, anamnesis of GO≥1 year) and 24 healthy volunteers. Corticosteroid therapy consisted of intravenous infusions of metyloprednisolone (MP) and subsequent treatment with oral prednisone (P). The serum samples were collected 24 h before MP, 24 h after MP, 12±2 days of treatment with prednisone and after the end of the corticosteroid therapy. The levels of soluble l -selectin and ICAM-1 in the serum were determined by the ELISA method. The statistical significance was estimated by the Mann-Whitney U-test. Serum l -selectin and ICAM-1 were significantly increased in patients with GO (respectively 1182±222 and 438±68 ng/ml) and in patients with Graves’ disease without ophthalmopathy (respectively: 1168±130 and 343±109) in relation to the controls. After MP treatment in corticosteroid-responsive patients (improvement in CAS ≤1) serum concentration of l -selectin and ICAM-1 decreased significantly and gradually increased during subsequent treatment with prednisone. In corticosteroid-responsive patients serum l -selectin was significantly higher before MP administration and after P treatment in relation to corticosteroid-resistant subjects. Conclusions: 1. Serum l -selectin and ICAM-1 were elevated in patients with active GO 2. Metyloprednizolone decreased levels of the studied adhesion molecules in corticosteroid-responsive patients with GO 3. Lack of clinical results in corticosteroid therapy in patients with a low starting l -selectin level would suggest the possibility of serum l -selectin estimation as a prognostic for immunotherapy efficacy.

The aim of this study was to evaluate lipid peroxidation and scavenging enzyme activity in streptozotocin-induced diabetes, and then to establish whether moderate doses of nonenzymatic antioxidant vitamin E play a role in the antioxidant defence system in diabetic pregnant rats and their offspring. The study group consisted of 30 normal female Wistar rats, which were given a single dose of streptozotocin (40 mg/kg) and were mated 7 days later. Subsequently, the diabetic animals were divided into two matched groups: the first supplemented with vitamin E (30 mg/100 g chow), and the other fed with a standard diet lacking vitamin E. Controls consisted of 15 pregnant rats. On the first day after delivery, the rats were decapitated and homogenates of maternal liver and uterus as well as neonatal lungs and liver were prepared. Then the following parameters were measured: malondialdehyde (MDA) concentrations in the homogenates and blood serum, glutathione (GSH) levels, the activity of CuZn-superoxide dismutase (SOD) and glutathione peroxidase (GPx), and glycaemia. The neonates of diabetic rats were smaller than the healthy ones and serum glucose concentration was markedly higher in the diabetic animals. MDA levels were significantly increased, whereas GSH, SOD and GPx were markedly diminished in the diabetic adult rats and their offspring in comparison to the control group. In the animals supplemented with alpha-tocopherol, MDA concentrations were significantly lower, GSH content and SOD activities were markedly elevated most tissues studied, whereas GPx remained unchanged. We conclude that, by monitoring the activity of selected scavenging enzymes, information on ongoing biological oxidative stress and thereby on the fetus/neonate status may be obtained. Our results suggest that diabetic pregnant rats and their neonates are exposed to an increased oxidative stress and that vitamin E supplementation may reduce its detrimental effects.

There have been some studies published recently which have suggested that L-selectin and/or other adhesion molecules could be the new markers for diabetes type 1 risk development in humans and animal models of the disease. The alterations of soluble L-selectin have been found not only in overt but also in the preclinical stage of disease development and were independent from the presence of ICA — a marker of ongoing autoimmunity, but associated with HLA related genetic predisposition to insulin-dependent diabetes mellitus (IDDM). The aim of our study was to evaluate the frequency of the L-selectin gene T668C mutation (from thymine to cytosine at position 668) resulted in F206L an amino acid substitution in patients with overt diabetes and their unaffected first degree relatives in comparison to the unselected control population. In the unaffected siblings of IDDM subjects we have observed a significantly higher frequency of the L-selectin gene T668C mutation in comparison to their relatives with type 1 diabetes and healthy controls. It was also shown that there is an association between T668C mutation and low HLA related risk of IDDM development, the highest frequency of F206L mutation in the EGF domain of L-selectin was observed in relatives with ‘protective’ HLA DQB1*0602 allele and nonDRB1*03-nonDRB1*04 haplotype, while in subjects with highest risk of IDDM haplotype the frequency of T668C mutation was similar to the controls. We would like to hypothesise that the T668C L-selectin gene mutation could have a (protective?) role in the development of IDDM, but further studies concerning their role in type 1 diabetes are needed.

The aim of the present study was to estimate whether a single bout of exhaustive exercise influences the glycogen and triglyceride (TG) content in red and white gastrocnemius muscle and in the liver of rats with experimental type 2 diabetes. Experiments were carried out on male Wistar rats fed from 8 to 11 weeks of age with isocaloric standard or high-fat diet (HFD) with a previous injection of low-dose of streptozotocin (STZ) or vehicle at 2 days of age (I, control group; II, HFD; III, STZ; IV, STZ + HFD). Group IV (STZ + HFD) represents a model of type 2 diabetes. Basal liver glycogen was markedly lower in all the studied groups compared to controls. Glycogen concentration after exercise fell significantly in the examined tissues in all groups in comparison to basal conditions. A significant TG accumulation in examined tissues was observed in all the studied groups in comparison to controls. Exercise decreased tissue TG content in all the groups, but it remained significantly higher in the experimental groups vs. control. We conclude that in this model of type 2 diabetes, a single bout of exercise reveals defective utilization of tissue carbohydrates and lipids.

Macrophages and T lymphocytes are the first cells to appear in pancreatic islets in the development of autoimmune diabetes. It has been suggested that cytokines released by monocytes/macrophages, including interleukin-1beta (IL-1beta), interleukin-12 (IL-12) and tumour necrosis factor-alpha (TNF-alpha) could have an initial role in islet B-cell damage. The aim of the present study was to estimate the effect of human insulin and nicotinamide on the levels of monocyte/ macrophage derived cytokines in the peripheral blood of humans at risk of Type 1 diabetes, and in patients with newly diagnosed Type 1 diabetes compared to healthy control subjects. The study was carried out on three groups of subjects: 20 first degree relatives of people with Type 1 diabetes (with two or more antibodies against pancreatic B-cell antigens); 22 patients with recent onset of Type 1 diabetes (duration of the disease 3-6 months); and 25 age- and sex-matched healthy subjects. Cytokine levels (IL-1beta, IL-12, and TNF-alpha) in the supernatants of whole blood cultures incubated with PHA alone (10 microg/ml), or PHA + human insulin (50 microg/ml), or PHA + nicotinamide (100 micromol/l) were quantified by ELISA. In the cultures with nicotinamide the concentration of IL-12 and TNF-alpha was significantly lower in the prediabetic group, diabetic patients, and the healthy controls than in the cultures with PHA only or with PHA + insulin. There were no significant differences in IL-1beta production in the cultures after incubation with the different stimuli in the studied groups and healthy controls. No significant influence of human insulin on macrophage/monocyte cytokines secretion in in vitro cultures of the peripheral blood was found. This suggests that nicotinamide could influence monocyte/macrophage function in peripheral blood by inhibiting production of IL-12 and TNF-alpha.

Summary L-selectin (CD62L) is a cell adhesion molecule which plays a key role in the initiation of leucocyte migration from blood vessels to sites of local inflammation. The aim of this study was to investigate T-lymphocyte expression of CD62L antigen and serum levels of soluble L-selectin (sL-selectin) in subjects with clinical and preclinical type I diabetes to determine whether they could provide surrogate markers for disease activity. CD62L selectin expression on memory T lymphocytes was studied by cytometric analysis in 22 patients with newly diagnosed type I diabetes, 20 first-degree relatives of patients with type I diabetes, 14 patients with Graves’ disease, and 22 healthy controls. sL-selectin levels were measured by enzyme-linked immunosorbent assay (ELISA) in enlarged groups of subjects in these categories, as well as in patients with long-standing type I diabetes, treated Graves’ disease and type II (non-insulin dependent) diabetes. L-selectin levels were also related to islet autoantibodies, human leucocyte antigen (HLA) genotype and L-selectin T668C gene polymorphisms. L-selectin expression on memory T lymphocytes was reduced in newly diagnosed diabetes and islet autoantibody positive siblings compared with controls. sL-selectin levels were significantly raised in newly diagnosed type I diabetes compared with controls, with intermediate levels in family members, both with and without islet autoantibodies, and in long-standing type I diabetes. Levels were also raised in patients with untreated Graves’ disease. Patients with type II diabetes had sL-selectin levels which did not differ from controls. sL-selectin levels correlated with the presence of diabetes-associated HLA alleles in both family members and controls; levels also fell with increasing age in family members. Multiple regression analysis showed that HLA genotype and age were independent determinants of sL-selectin levels. sL-selectin levels are raised at the time of diagnosis of type I diabetes and Graves’ disease and appear to be modulated by disease activity, but levels are determined predominantly by HLA-associated genetic susceptibility and age. sL-selectin may provide a late marker of autoimmune destruction of islets and sequential measurement may be useful in monitoring disease activity and the effect of interventions preceding type I diabetes.

The aim of our study was to analyse the effect of chronic hyperglycaemia on lipid peroxidation and scavenging enzyme activity in pregnant animals and their offspring supplemented and not supplemented with vitamin E - a natural antioxidant. Thirty pregnant female Wistar rats were used in our experiments. Diabetes was induced on day 7 of pregnancy using a single does of streptozotocin (40 mg/kg). Diabetic animals were divided into two equal groups: vitamin E supplemented and those fed with standard diet. Our controls consisted of 15 healthy rats. On day 1 after delivery homogenates of maternal liver and uterus as well as neonatal lungs and liver were prepared. Then the following parameters were measured: malondialdehyde (MDA) concentrations in the homogenates and blood serum, glutathione (GSH) levels, the activity of CuZn superoxide dismutase (SOD) and glutathione peroxidase (GPx) (Bioxytech, France). Statistical analysis was performed using Mann-Withney U test. The neonates of diabetic rats were smaller than those from healthy rats and serum glucose concentration was markedly higher in diabetic animals, both in mothers and neonates. MDA levels increased significantly, whereas GSH content and SOD as well as GPx activities were markedly diminished in diabetic pregnant rats and their offspring in comparison with the control group. In animals supplemented with tocopherol, MDA concentrations declined significantly, GSH contents and SOD activities were markedly elevated in almost all types of tissues studied, whereas glutathione peroxidase remained suppressed. Our results suggest that diabetic pregnant rats and their neonates are exposed to oxidative stress (OS), but vitamin E supplementation could in part reduce the imbalance between uncontrolled reactive oxygen species generation and scavenging enzyme activity, and may potentially serve as a useful prophylactic factor against OS development

There is increasing evidence that CD3 + cells bearing gammadelta T-cell receptor (represent the minor subpopulation of the T-cells in the peripheral blood in humans) are involved in autoimmunity development. Gammadelta T-cell receptor (TCR)+ /CD8+ T-cells have been recently found to play a critical role in the pathogenesis and prevention of autoimmune diabetes in the animal model. The aim of the present study was the estimation the gammadelta T-cell subpopulation levels in the peripheral blood of subjects with preclinical and overt type 1 diabetes and their possible associations with the humoral immunity, metabolic parameters and pancreatic B-cells function. The study was carried out in three groups of subjects: 26 first degree relatives of type 1 diabetes patients (prediabetics) with the combinations of autoantibodies against pancreatic B-cells (ICA, GADA, IA-2A, IAA), 22 patients with a recent onset of type 1 diabetes and age and sex-matched 24 healthy volunteers (control group). A decrease was observed in the absolute numbers and percentages of gammadelta+ /CD8+ and gammadelta+ /CD8- T-cell subpopulations in peripheral blood in the prediabetics with the impaired first phase of insulin secretion in comparison to relatives with autoantibodies but still with normal B-cells function, patients with clinical diabetes and healthy controls. In conclusion, the study suggests that the gammadelta T-cells play an important role in the development of insulin-dependent diabetes mellitus (IDDM). It is possible that their levels in the peripheral blood could be an additional marker of preclinical detection of the disease, but further prospective studies in high risk of IDDM subjects are needed.

There is mounting evidence that the imbalance between Th1 and Th2 lymphocyte subsets plays a key role in the development of autoimmune diabetes in NOD mice, but it is also possible in humans. The aim of the present study was the estimation of in vitro production of Th1 (INF-gamma, IL-2) and Th2-derived (IL-4, IL-10) cytokines by peripheral blood in ICA and GADA positive first degree relatives of Type-I diabetes patients, since they could represent primary events triggering an immune-mediated islets destruction. The study was performed in 25 subjects at risk of insulin-dependent diabetes and 21 age- and sex-matched healthy controls. Cytokine levels in supernatants of whole blood cultures with PHA (10 microg/ml) were quantified by ELISA. We observed a lower concentration of IL-4 in culture supernatants in ICA and GADA positive relatives as compared with the control group, both at 48 h and at 72 h of incubation. Similarly, in the prediabetic group, lower IL-10 levels at 48 and 72 h of culture were found. We did not observe statistical differences in in vitro production of IL-2 and INF-gamma by peripheral blood in high risk diabetes mellitus subjects and healthy controls. In subjects at increased risk of Type-I diabetes, levels of IL-4 positively correlated with those of IL-10. There were negative correlations between IL-10 concentration after 48 h of incubation and levels of HbA1C. In conclusion our study has shown decreased IL-4 and IL-10 production, but normal secretion of Th1-derived cytokines by peripheral blood of prediabetic humans. This could suggest that the early stage of autoimmune process in Type-I diabetes in humans is associated with decreased function of Th2-cells rather than overactivation of Th1 subset in the peripheral blood.

The aim of this study was to investigate whether sulodexide treatment is capable of influencing urinary albumin excretion rate (UAER) in insulin-dependent diabetes mellitus patients (type I) with micro- or macroalbuminuria. A total of 14-inpatients (seven with micro and seven with macroalbuminuria) were enrolled and were treated first intramuscularly with a 60 mg vial of sulodexide/day for 10 days, and then orally with 25 mg capsules twice a day for 21 days. UAER was estimated before starting treatment (T0), after the i.m. treatment phase (T1) and at the end of the oral treatment (T2). No statistically significant differences in hematochemical and coagulative parameters were registered after treatment, with respect to basal values. On the contrary, a marked decrease in UAER mean values was registered at the end of both the parenteral and the oral treatment periods (T0: 349.9 mg/24 h, range 80-820; T1: 237 mg/24 h, range 7-620; T2: 91.4 mg/24 h, range: 2-306). All the differences were statistically significant (P < 0.001) versus baseline. At T2, a normalisation of UAER was observed in three microalbuminuric and in two macroalbuminuric patients, and a remarkable decrease was found in additional four and five patients, respectively. UAER was found to be still significantly lower than at baseline after 6 weeks of follow-up. This preliminary study suggests that sulodexide is effective in reducing UAER in type I patients with diabetic nephropathy.

Cytokines secreted by antigen presenting cells, lymphocytes T and pancreatic beta cells are considered as the major mediators in the pathogenesis of IDDM. It has been suggested that cytokines released by macrophages/monocytes could have an initial role in beta-cell damage. The aim of the present study was the estimation of in vitro production of macrophage-derived cytokines: IL-1 beta, TNF-alpha, IL-12 by peripheral blood in high risk IDDM first degree relatives, since it could reflect early events leading to the development of type 1 diabetes in humans. The study was performed in 25 high risk IDDM subjects and 21 age and sex-matched healthy controls. IL-1 beta, TNF-alpha and IL-12 concentrations in supernatants of whole blood cultures with PHA (10 micrograms/ml) were quantified by ELISA. In the ICA positive relatives of IDDM subjects levels of IL-12 were significantly higher as compared with the control group, both at 48 h (p < 0.02) and at 72 h (p < 0.05) of incubation and positively correlated with TNF-alpha and IL-1 beta (R = 0.46, p < 0.02 and R = 0.32, p < 0.05). We did not observe statistical differences in in vitro production of TNF-alpha and IL-1 beta between the study groups. In conclusion we suggest that our findings support the hypothesis, that IL-12 is involved in the pathogenesis of human IDDM. If the involvement of Th1 cells is confirmed in the destruction of islet beta-cells, it is possible that IL-12 antagonists will have a role in the future prevention of insulin dependent diabetes mellitus.

The aim of our study was to investigate the incidence of thyroid autoantibodies (TA) in the endemic region of the north-eastern Poland. The mean titres of ATMA and TGAb were measured in 1508 randomly chosen persons aged 3 to 68 yrs. ATMA or TGAb were positive in 17% of the population studied. Frequency and mean titres of TA were observed to increase with age. Autoantibodies were more frequently noted in persons with parenchymatous goiter. There was no correlation between the incidence of autoantibodies and goiter enlargement, however, the highest percentage of TGA was noted in people with large thyroid (III). TA were also found in 10% of the subjects without goitre. Our results do not confirm the hypothesis that thyroid autoantibodies play a key role in the pathogenesis of endemic goiter.

Type 1 diabetes is believed to be a Th1 lymphocyte–mediated disease, and both environmental and genetic factors play a role in its pathogenesis. It was recently found that interleukin (IL)-18 acts as a proinflammatory cytokine and, in synergy with IL-12, promotes development of Th1 lymphocyte response by induction of γ-interferon production. The aim of our study was to evaluate the frequency of known polymorphisms in the IL-18 promoter in patients with type 1 diabetes in comparison with healthy control subjects, since higher levels of IL-18 were recently reported in the subclinical stage of type 1 diabetes. We studied two recently described single-nucleotide polymorphisms of the promoter of IL-18 gene at the position −137 and −607, which have been suggested to cause differences in transcription factor binding and have an impact on IL-18 gene activity. The genotype distribution differed significantly between patients with type 1 diabetes and control subjects. The difference reflected an increase in the GC genotypes and a decrease in GG genotypes at position −137 in the promoter of IL-18 gene. AA genotype at position −607 was found only in the control group. The results also demonstrated that the contribution of −137GC genotypes to genetic susceptibility to type 1 diabetes differs depending on the combination of IL-18 promotor gene haplotypes. Our study suggests the first evidence of an association between type 1 diabetes and polymorphisms in the promoter of IL-18 gene.

The number of patients with chronic kidney disease-CKD is still growing. Overweight and obesity present also an important problem of world public health. However, there are not many data showing possible association between obesity and incresing risk of development of renal failure recently it has been demonstrated that in obese patients secondary focal segmental glomerulosclerosis and glomerular hypertrophy appear more frequently. The aim of this study was to estimate glomerular filtration rate-GFR in patients with normal serum creatinine concentration undergoing primary angioplasty according to body mass index. The study included 1413 patients udergoing primary angioplasty for acute myocardial infarction. The following parameters were assessed: age, gender, family history of cardiovascular disease, risk factors of cardiovascular disease (hypertension, diabetes mellitus, obesity etc.), previous myocardial infarction, pre-existing heart failure, treatment given, localization of infarct, coronary stenting, serum creatinine before angioplasty, cholesterol, LDL, HDL, triglycerides, glucose, blood pressure. Of a total of 1413 patients, 1337 (94.62%, 943 M, 394 F) had correct serum creatinine concentration (below 1.5 mg/dl for men, below 1.2 mg/dl for women). Glomerular filtration rate was calculated from serum creatinine levels by using the simplified Modification of Diet in Renal Disease Study formula--MDRD, Cockcroft-Gault equation and Jeliffe formula. An average value of GFR in study group was 79.94 +/- 24.51 ml/min (Cockcroft-Gault equation), 73.02 +/- 21.96 ml/min (Cockcroft-Gault adjusted to weight), 90.37 +/- 25.1 ml/min (MDRD equation) and 77.67 +/- 21.65 ml/min (Jeliffe formula). A significant lower serum creatinine levels and GFR (assessed by 3 formulas and Cockcroft-Gault using adjusted weight) were observed in women group. In the whole study group (with normal serum creatinine levels) substantial correlation was found between age and serum creatinine concentration (r = 0.13, p0.001), GFR (MDRD, r = -0.37, p0.001, Cockcroft-Gault, r = -0.62, p0.001, adjusted to weight r = -0.64, p0.001, Jeliffe r = -0.61, p0.001) and also between BMI and GFR (MDRD r = 0.28, p0.001, Cockcroft-Gault, r = 0.31, p0.001, adjusted to weight r = 0.08, p0.001, Jeliffe r = 0.341, p0.001), but not with serum creatinine concentration (r = 0.03, p = 0.3). In patients with normal serum creatinine levels percentage of patients with GFR below 60 ml/min ranges from 4.79% up to 30.74%. In patients with higher BMI, higher GFR may be partially caused by glomerular hyperfiltration. Overweight or obesity are significant, but potentially changeable risk factors for development of chronic renal failure. However, chronic kidney disease is one of the complications of obesity.

Gestational Diabetes Mellitus (GDM) comprises different forms of glucose metabolism disturbances with first recognition during pregnancy. There are a number of publications that have suggested that diabetes with onset during pregnancy is not a monogeneous disease and apart from "classical" form of GDM, which precedes type 2 diabetes development, MODY 2 diabetes, caused by monogenic defect of glucokinase gene, is relatively frequent "subtype" of gestational diabetes. The aim of our study was to estimate the risk of diabetes mellitus development, including MODY 2 diabetes, between 6 months - 10 years after delivery in 225 women with gestational diabetes. Gucokinase gene mutations and polymorphisms were performed by direct sequencing of DNA. In the present study it was shown that the frequency of glucokinase gene mutation is 6.7% in the Polish population of gestational diabetic women and 17.8% of new onset or persistant diabetes recognised during 5 years after pregnancy could be a result of this mutation. We have also observed that risk of type 2 diabetes development is about 50% in the next 5 years after delivery in women with gestational diabetes and is associated with higher levels of BMI during or after delivery and with clinical and biochemical features of insulin resistance (high values of WHR abd HOMA-R). Moreover, our study suggests that c.1253+8 C--T polymorphism in intron 9 of glucokinase gene could have a role in predisposition to type 2 diabetes in women with gestational diabetes. In summary, our results suggest that, because of high costs and time-consuming methods of genetic studies, the investigations of glucokinase gene mutations should be concentrated in women with gestational diabetes without clinical and biochemical features of insulin resistance, but with family history of diabetes in two generations.

Insulin resistance is the underlying metabolic abnormality in the metabolic syndrome. The low-grade chronic inflammation may be associated with metabolic risk factors and atherogenesis. The aim of our study was to establish the link between the metabolic syndrome, as defined by the National Cholesterol Education Program (NCEP) criteria, and insulin sensitivity, serum adiponectin, and parameters of chronic inflammation in young subjects. The group of 223 subjects (mean age, 25.86 ± 5.49 years; body mass index, 28.04 ± 6.91 kg/m2) was studied. Oral glucose tolerance test, euglycemic hyperinsulinemic clamp, and estimation of serum adiponectin and proinflammatory factors were performed. The NCEP-defined metabolic syndrome was present in 49 subjects (21.97%). The higher the number of NCEP criteria fulfilled was, the bigger were the decrease in insulin sensitivity (P < .0001) and adiponectin (P < .0001) and the increase in fasting and postload insulin (both Ps < .0001), C-reactive protein (P < .0001), interleukin 18 (P < .0001), interleukin 6 (P < .0001), and soluble tumor necrosis factor–α receptors sTNFR1 (P < .0001) and sTNFR2 (P < .0001) observed. Multiple regression analysis revealed that adiponectin and inflammatory factors predicted NCEP score independent of insulin sensitivity (all adjusted β values between .16 and .32, all Ps < .01). Young subjects with metabolic syndrome demonstrate an increased inflammatory response and lower adiponectin concentration.

Up-to-date observations in the regions where effective food iodine supplementation was introduced confirm a significant decrease in goiter incidence. However, in some regions the incidence of goiter remains unchanged or even the increase in the frequency of autoimmunological diseases of the thyroid like, Grave's-Basedov disease and chronic/subacute lymphocytic thyroiditis is observed. The purpose of the study was to evaluate the prevalence of autoimmunological diseases of the thyroid in children aged 6-13 years in the Polish population after 5 years of obligatory iodine supplementation program. The study included 480 school children from 4 elementary schools chosen randomly in the city of Białystok and 120 patients at the same age treated due to goiter with KI and/or thyroxin for minimum 12 months in the Regional Specialist Outpatient Clinic of Endocrinology. All children underwent physical examination with palpation of goiter and USG of the thyroid. Iodine concentration was assessed in the morning urine by the catalytic method of Sandell-Költhoff. The concentrations ofTSH and antibodies against TSH, thyroid-peroxidase (TPO) and thyroglobulin (Tg) were also determined. The mean concentrations of anti-Tg antibodies were statistically significantly higher in children with goiter in comparison with children with the thyroid gland within the norm (155.8 IU/ml vs. 24.4 IU/ml, p = 0.045). In children with goiter the incidence of'positive' titre of anti-Tg antibodies was 4-fold as high as in children without goiter (33.3% vs. 8.7%, p = 0.0147). High incidence of antibodies against thyroglobulin in the population of children with goiter and a positive correlation between the titre of anti-Tg antibodies and the size of the thyroid in this group suggest a significant role of autoimmunological disorders in the pathogenesis of goiter in the studied population.

Tumor necrosis factor-alpha (TNF-alpha) system is potentially involved in the development of insulin resistance during pregnancy. Plasma concentrations of TNF-alpha and its soluble receptors sTNFR-1 and sTNFR-2 were measured in 80 patients with gestational diabetes (GDM) (mean age 29.0 +/- 4.9 years) and 30 pregnant women with normal glucose tolerance (NGT) (mean age 28.2 +/- 6.0 years). We found that patients with GDM had significantly higher levels of TNF-alpha in comparison to NGT women (1.71+/- 0.92 vs. 1.27 +/- 0.42 pg/ml, p = 0.0175). The differences remained statistically significant after adjusting for BMI (p = 0.027). Plasma levels of sTNFR-1 and sTNFR-2 were only slightly higher in patients with GDM (2.83 +/- 0.79 ng/ml vs. 2.55 +/- 0.99 ng/ ml, p = 0.057 and 7.46 +/- 2.21 ng/ml vs. 6.83 +/- 1.46 ng/ml, p=0.206, respectively). In the group with GDM TNF-alpha concentrations correlated with sTNFR-1 (r = 0.444, p = 0.00008), sTNFR-2 (r = 0.364, p = 0.0016) and with C-peptide concentrations (r = 0.318, p = 0.016), whereas in women with NGT - only with triglyceride levels (r = 0.50, p = 0.024). Multivariate linear regression analysis revealed that early pregnancy BMI was the most predictive indicator of TNF-alpha concentrations in GDM women (p=0.008). In NTG group triglyceride concentrations, as well as BMI in early pregnancy and at the time of sampling were significant predictors, explaining together 62% of the variance in TNF-alpha concentration. In conclusion, increased TNF-alpha concentrations in women with GDM class G1 indicates its contribution to the development of insulin resistance during pregnancy, but the lack of the differences in sTNFR concentrations between the groups studied suggests only moderate TNF-alpha system activation in relatively slim patients treated with diet.

Proper diet with regard to quantity and quality of meals is of vital importance for normal development and functioning of the organism. There are many proofs that environmental factors play an important role in the pathogenesis of goiter. Iodine deficit in diet is best known of all factors contributing to goiter. Deficit of other elements like, iron, selenium, copper and zinc is also essential. The purpose of this study was to evaluate the influence of chosen environmental factors, i.e., iron and trace elements of selenium, zinc and copper--essential for the thyroid functioning on the development of goiter in school children aged 6-13 years with normal ioduria in the Polish population.In 2002, the study was performed in 4 elementary schools chosen randomly in Białystok and in the Children's Outpatient Clinic of Endocrinology of the Specialist Regional Hospital. The study included 400 children aged 7-13 years from schools and 120 patients at the same age treated with KJ and/or tyroxine for minimum 12 months due to goiter in the Out-patient Clinic of Endocrinology. Basing on the assessment of the thyroid size as well as the criteria of WHO from 1997 year for body surface and sex, children were divided into 2 subgroups: with goiter and the thyroid gland within the norm. Children aged 9-11 years were qualified and chosen from subgroups to further examinations. In both subgroups, blood samples were taken to determine concentrations of iron, selenium, copper and zinc.The mean concentration of selenium in the blood was statistically significantly lower in children with goiter in comparison with children with the thyroid gland within the norm (44.4 +/- 7.8 microg/L vs. 49.2 +/- 9.1 microg/L, p = 0.044) in the study population of school children and the Outpatient Clinic of Endocrinology. No differences of serum iron concentrations were observed in children with goiter and with the thyroid gland within the norm. However, nearly the half (45.5%) of patients with the lower serum concentration of iron (60 microg/dL) had goiter despite average 22-month therapy with KJ and/or tyroxine.Observed, in spite of proper iodine prophylaxis, 7% rate of goiter occurring in school children suggests other than iodine deficiency factors that influence goiter development. The study proved that the low concentration of iron and/or selenium deficit found in the serum of children with goiter in spite of their treatment with KJ and/or tyroxine may be additional factors influencing the effectiveness of this treatment.

Objective: Serum concentrations of soluble tumor necrosis factor-α (TNF-α) receptor 2 (sTNFR2) are associated with insulin resistance. In a recent study, we provided evidence for the existence of a biologically active form of sTNFR2 produced by alternative splicing (DS-TNFR2). We aimed to evaluate whether this circulating DS-TNFR2 is associated with insulin action in humans. Design and methods: Real time PCR (light cycler technology) evaluated DS-TNFR2 expression in monocytes. DS-TNFR2 was measured using a monoclonal antibody against an epitope present in TNFR2 (first 14 residues of the juxtamembrane region) but predicted to be absent in soluble proteolytic cleavage-produced TNFR2. Insulin sensitivity was measured using euglycemic hyperinsulinemic clamp (n = 76) and homeostatic model of assessment (HOMA) value in a replication study of 223 subjects. Results: Real time PCR confirmed gene expression of DS-TNFR2 in monocytes from healthy subjects. A significant and positive association was found between serum DS-TNFR2 concentration and insulin sensitivity (P = 0.032, n = 76). This association was most significant in subjects with normal glucose tolerance (r = 0.44, P = 0.002). The subjects in whom DS-TNFR2 was detectable were more insulin sensitive than those with undetectable DS-TNFR2 (42.12±22.08 vs 31.71± 16.95 μmol × kg−1 × min−1, P = 0.039). DS-TNFR2 was inversely associated with body mass index, waist-to-hip ratio, systolic and diastolic blood pressure, fasting serum glucose, serum triglycerides and serum uric acid concentration and with the HOMA value (P = 0.03) in the replication study. Circulating DS-TNFR2 declined with increased number of components of the metabolic syndrome. Conclusion: Native sTNFR2 and DS-TNFR2 show opposite associations with insulin action. DS-TNFR2 might play a role as a counterpart of the proinflammatory environment associated with insulin resistance.

Growth factors--IGF-1 and TGF-beta1 are well documented factors regulating proliferation of follicle cells of the thyroid in many experiments in vitro. It has been proved so far that IGF-1 stimulates cellular mitogenesis of thyrocytes, whereas TGF-beta1 inhibits proliferation of follicle cells of the thyroid in experimental conditions.The aim of the study was to evaluate the correlation between serum concentrations of IGF-1 and TGF-beta1 and the size of the thyroid in children with normal ioduria.In 2002, the study was performed in 4 elementary schools chosen randomly in Białystok and in the Children's Out-patient Clinic of Endocrinology of the Specialist Regional Hospital. The study included 480 children aged 7-13 years from schools and 120 patients at the same age treated with KJ and/or thyroxine for minimum 12 months due to goiter in the Out-patient Clinic of Endocrinology. All children underwent physical examination with palpation of goiter and USG of the thyroid. Iodine concentration was assessed in the morning urine by the catalytic method of Sanedell-Kolthoff. In the second part of the examination, basing on the assessment of the thyroid size as well as the criteria of WHO from 1997 year for body surface and sex, children were divided into 2 subgroups: with goiter and the thyroid gland within the norm. Children aged 9-11 years were qualified and chosen from subgroups to further examinations. In both subgroups (with goiter and normal thyroid gland) blood samples were taken to determine concentrations of TSH, IGF-1, TGF-beta1.The mean values/median of IGF-1 concentration were statistically significantly increased in children with goiter in comparison with children with a normal thyroid (436.2 vs. 343.8 ng/ml, p=0.047). The mean values / median of TGF-beta1 concentration were statistically significantly decreased in children with goiter when compared to children with the thyroid gland within the norm (17.8 vs. 23.9 ng/ml).The significantly lower concentration of TGF-beta1 in the serum of children with goiter in comparison with the values in children with normal size of the thyroid gland and a positive correlation between the concentrations of IGF-1 and the size of the thyroid (after excluding the influence of age and body surface) seem to confirm a vital role of IGF-1 and TGF-beta1 in the pathomechanism of goiter.

The evaluation criteria of the enlarged thyroid gland becomes a key problem when analyzing the prevalence of goiter in the population of school children. The review of the literature and own experience indicate lack of a consensus concerning this problem among researchers. Similarly, in the report of WHO in the year 2001 concerning the problem of iodine deficit and goiter endemia, experts did not present universal referential values of the thyroid size in ultrasonography for children population aged 6-15 years living in the regions of proper iodine supply in the diet, thus suggesting the necessity of working out regional norms.The aim of the study was to evaluate the prevalence of goiter in children aged 6-13 years from schools chosen randomly in Białystok with proper iodine supplementation in the study population and to estimate the usability of referential values applied in the assessment of the thyroid size.In the year 2002, the examination was carried out in 4 elementary schools chosen randomly from Białystok. A total of 480 children aged 6-13 years were included in the study. All children were examined physically with palpation assessment of the thyroid size and had USG of the thyroid. The concentration of iodine was measured in the morning urine. The blood samples were collected to determine hTSH concentration.In palpation, with regard to WHO criteria of the year 2001, the prevalence of goiter was 6.8% in the study population. Applying WHO criteria of 1994 in palpation assessment of goiter increased this percentage up to 18.2%. When using WHO criteria for body surface of 1997 to evaluate goiter by USG, its percentage equaled 7%, whereas taking into consideration referential values for child's age doubled its percentage up to 13.5%. The percentage of goiter increased up to 45.5%, when referential values introduced by Gutekunst et al. were applied.When evaluating the thyroid size in ultrasonographic diagnostics of goiter in children aged 6-13 years, it seems more purposeful to apply referential values of the thyroid size regarding the body surface and sex (manifesting a child's actual physical development) than to use the norms concerning the calendar age. The prevalence of goiter amounting 7% in the population of school children in spite of adequate iodine prophylaxis suggests other than iodine deficit, factors causing goiter in this population.

Leptin is a protein hormone mainly produced by the adipocytes. Apart from its autocrine role within the placenta in humans, plasma circulating leptin contributes to the endocrine function. Leptin levels may serve as an index of metabolic and energy balance in pregnancy. Recent reports have shown a positive correlation between leptin concentrations and plasma levels of glycated haemoglobin (HbA(1c)) in patients with gestational diabetes mellitus (GDM).The aim of the present study was to evaluate leptin levels after delivery in GDM and normal glucose tolerance (NGT) women.Plasma leptin concentration and insulin, c-peptide and glycated haemoglobin were measured in both. NGT women and in patients with a history of GDM in all patients total LDL - and HDL cholesterol concentrations were estimated. We also calculated the anthropometric parameters of the mother and birth weight in both groups.The plasma leptin concentration after delivery was not different in patients with GDM in comparison with the NGT individuals.We concluded that in patients with GDM and normal BM1 the postpartum leptin level was not different in comparison with the NGT patients.

Circulating monocyte-platelet aggregates can release procoagulant, oxidative and mitogenic factors, thereby contributing to arterial thrombosis. The aim of our study was the estimation of heterophilic leukocyte-platelet aggregates in patients referred for coronary angiography, dependent on the degree of coronary stenosis and the disturbances of carbohydrate metabolism. Flow-cytometric analysis was performed in 50 consecutive patients with positive exercise test (age 54.2 +/- 6.4 years): 27 with normal glucose tolerance, 7 with impaired glucose tolerance and 16 with type 2 diabetes, and in 16 healthy subjects (age 44.8 +/- 14.1 years). We found that patients with coronary heart disease had increased leukocyte-platelet aggregate formation in comparison to the controls (the percentage of monocyte-platelet aggregates 47.5 +/- 23.0 vs 25.7 +/-12.8, p = 0.003, mean fluorescence intensity (MIF) 187.6 +/- 117.2 vs 79.3 +/- 42.8, p = 0.002, the percentage of granulocyte-platelet aggregates 20.7 +/- 10.4 vs 17.0 +/- 3.6, p = 0.009, MIF 64.2 +/- 41.3 vs 40.9 +/- 6.3, p = 0.008). The highest percentage of heterophilic aggregates was observed in patients with 1- and 2-vessel disease and those with "clean" vessels. In diabetic patients the percentage and MIF of granulocyte-platelet aggregates were decreased in comparison to the subjects with normal glucose tolerance (16.7 +/- 7.2 vs 22.8 +/- 9.8, p = 0.03 and 44.3 +/- 10.8 vs 74.4 +/- 48, p = 0.009, respectively). There was no increase in glycoprotein CD14 expression in any of the group studied. We found a positive correlation between the percentage of monocyte-platelet aggregates and fasting insulin level (r = 0.369, p = 0.04) and a negative correlation between MIF of monocyte-platelet aggregates and HDL level (r = -0.459, p = 0.012), between MIF CD14 and HDL level (r = -0.435, p = 0.02), and between the percentage of granulocyte-platelet aggregates and postprandial glycaemia (r = -0.4117, p = 0.03). We concluded that: 1. the patients with "clean" vessels represent a group of high atherothrombotic risk. 2. the patients with minimal coronary stenosis may benefit from anti-inflammatory and antiplatelet treatment.

Apoptosis plays an important role in the pathogenesis of autoimmune thyroid diseases. Soluble form of Fas (sFas) appeared as reliable markers of inflammation activity in rheumatic autoimmune diseases. The aim of the study was an estimation of sFas in patients with Graves' disease with ophthalmopathy during treatment with corticosteroids to assess their potential as a guideline of immunosuppressive therapy.We detected serum Fas in three groups of subjects: 25 patients with clinical symptoms of ophthalmopathy (OG)(CASor = 4, anamnesis of OGor = 1 yr), 18 patients with Graves' disease without ophthalmopathy (ChG) and 14 healthy volunteers. Corticosteroid therapy consisted of intravenous infusions of methylprednisolone (MP) and subsequent treatment with oral prednisone. The serum samples were collected 24 hours before MP, 24 hours after MP, 14 days of treatment with prednisone and after the end of the corticosteroid therapy. The levels of soluble Fas in the serum were determined by the ELISA and TSH receptor antibodies by RIA method.sFas concentration was significantly increased in patients with OG. We found a positive correlation between sFas and CAS and a negative correlation with duration of OG. During corticosteroid treatment there was no difference in sFas between responders and non-responders.sFas is a potent surrogate marker of inflammatory process activity in Graves' ophthalmopathy, however it has no prognostic value in the prediction of the immunotherapy efficacy.

The aim of our study was the estimation of platelet sodium-proton exchanger activity and platelet pro-coagulant activity, expressed as the availability of platelet factor 3 (PF3) and thrombin generation, in 83 type 2 diabetic patients (mean age 56.7 +/- 7.8 years) and 40 healthy subjects (mean age 54.4 +/- 6.2 years). Thrombin generation was measured in platelet rich plasma, using a chromogenic substrate S-2238. The availability of PF3 was estimated in platelet rich plasma, platelet poor plasma and platelet filtrated plasma, to assess procoagulant activity connected with platelets and cell derived microparticles, shedding upon activation (according to Jy and Horstman). The activity of platelet Na+/H+ exchanger was measured using an optical swelling assay. We found that the activity of PF3 and phospholipid dependent thrombin generation were significantly higher in diabetic patients, irrespective of their vascular complications and metabolic control. The highest increase of PF3 activity was observed in platelet poor (p0.0001) and platelet filtrated plasma (p0.000001). Na+/H+ exchange rate was significantly higher in diabetic patients in comparison to the controls (4.29 +/- 0.71 x 10(-3)/s vs 3.21 +/- 0.64 x 10(-3)/s, p0.00001). There was also a positive correlation between Na+/H+ exchanger activity and PF3 activity in all plasma fractions. Our results suggest that increased thrombin generation, enhanced platelet Na+/H+ exchanger activity and raised PF3 availability, connected mainly with cell derived microparticles, may enhance the risk of vascular damage in type 2 diabetic patients.

Insulin resistance is a key element of metabolic syndrome, which includes disturbances of glucose tolerance, obesity, hypertension, coronary heart disease dyslipidemia and many other defects. An important problem in scientific research is precise measurement of insulin sensitivity. The method considered "the gold standard" is glucose clamp, however, it is difficult to apply this method in large studies. Therefore, simple indices of insulin resistance are proposed. It remains unclear whether those indices are able to reflect changes occurring during insulin-sensitizing intervention. The aim of the present study was to assess the use of indirect indices for the changes in insulin sensitivity during exercise training and to compare those indices with results derived from clamp. Fourteen obese normoglycemic women participated in 12-week exercise training program, which included exercise performed on a bicycle ergometer, 5 days a week for 30 minutes. Insulin sensitivity (M/FFM value) before and after training was measured with hyperinsulinemic euglycemic clamp technique. Simple indices of insulin resistance were also assessed: fasting plasma insulin (INS), logarithm INS (log [INS]), homeostasis model assessment (HOMA), logarithm HOMA (log [HOMA]) and quantitative insulin sensitivity check index (QUICKI). Before training, all those indices were markedly related to M/FFM. After training, an increase in M/FFM was observed. None of the examined indices markedly changed after training. There was no correlations between changes of evaluated indices and in M/FFM during training, and no relationships of those parameters after training. Our study indicates that simple indices are not able to reflect changes occurring during insulin-sensitizing intervention.

It was recently suggested that genetic factors could play a major role in the development of Graves’ disease (GD). The aim of the present study was to evaluate the frequency of the c.721G3 A polymorphism and the c.1405A3 G polymorphism of the intercellular adhesion molecule 1 (ICAM-1) gene in subjects with GD compared with that in healthy controls, because ICAM-1 was found to play a key role in lymphocyte infiltration into the thyroid gland and the concentration of the soluble form of ICAM-1 correlates significantly with the clinical activity and treatment status in GD. We have analyzed the association of ICAM-1 polymorphisms with the age at onset of GD and the presence of ophthalmopathy. In a group of 235 patients with GD and 211 healthy controls we have shown that polymorphism at position c.721G3 A is associated with an earlier age of GD onset and that the c.1405A3 G polymorphism of the ICAM-1 gene could predispose to Graves’ ophthalmopathy. This suggests that G241R and K469E amino acid substitutions in the ICAM-1 molecule could influence the intensity/ duration of the autoimmunity process and the infiltration of orbital tissues. It could be speculated that therapy that modulates ICAM-1 function may delay the onset and/or prolong the remission and/or have an influence on clinical manifestations of GD. (J Clin Endocrinol Metab 88: 4945– 4949, 2003)

Insulin resistance is a key factor in the pathogenesis of impaired glucose tolerance (IGT) and type 2 diabetes and is also associated with greater risk for cardiovascular disease. Insulin resistance is more common in obese individuals and is considered to be the link between obesity and IGT and diabetes. The aim of the present study was to assess insulin resistance in obese subjects with IGT. We examined 57 subjects with marked overweight or obesity (BMI27.8 kg x m-2), 27 with IGT and 30 with normal glucose tolerance (NGT), assessed by an oral glucose tolerance test, according to WHO criteria. Thirty lean (BMI25 kg x m-2) healthy subjects served as a control group. Anthropometric and biochemical parameters were measured. Insulin sensitivity was evaluated with hyperinsulinemic euglycemic clamp technique. Subjects with IGT had higher levels of glucose, insulin, non-esterified fatty acids and glycated hemoglobin than obese with NGT, all those parameters were also higher in both obese groups in comparison to controls. We showed significant differences in insulin sensitivity between the studied groups, an index of the whole-body glucose uptake was decreased in both obese groups in comparison to controls, and it was also lower in IGT than in obese NGT group. We observed marked negative correlations between insulin sensitivity and estimated anthropometric and biochemical parameters. Our study indicates that insulin resistance is an important factor determining a deterioration of glucose tolerance in subjects with overweight and obesity.

Type 2 diabetes is characterized by impaired insulin secretion and insulin resistance. Both genetic and environmental factors are thought to play a role in the pathogenesis of this disease. Obesity is associated with increased risk of type 2 diabetes, but some metabolic abnormalities may also be present in lean, predisposed subjects. The aim of the present study was to assess insulin sensitivity in lean normoglycemic offspring of parents with type 2 diabetes.We examined 17 lean offspring (BMI25 kg x m-2) whose parents had type 2 diabetes, and 17 age, sex and BMI-matched subjects without family history of diabetes as controls. Anthropometric and biochemical measurements, oral glucose tolerance test (OGTT), and euglycemic hyperinsulinemic clamp were performed.The subjects did not differ in anthropometric measurements, and all had normal glucose tolerance according to WHO criteria. Offspring of type 2 diabetic parents were markedly more hyperinsulinemic (p0.05) and insulin resistant (p0.005). The ratio of the increase in insulin to the increase in glucose during the first 30 minutes of the OGTT, a crude index of first-phase insulin secretion, did not differ between groups. Insulin sensitivity was negatively related to fasting plasma insulin (r=-0.67, p0.001) and non-esterified fatty acids (r=-0.43, p0.02).Insulin resistance is present even in young lean subjects at high risk to develop type 2 diabetes. Our data suggest that insulin resistance may be a primary abnormality in the pathogenesis of this disease.

Background Interleukin 8 (IL-8) is a cytokine with atherogenic properties. In vitro studies revealed that it is produced and secreted by human adipocytes. We recently reported that plasma IL-8 is increased in obese subjects with normal glucose tolerance (NGT). The aim of the present study was to measure plasma IL-8 concentrations in subjects with impaired glucose tolerance (IGT). Methods A total of 44 subjects with marked overweight or obesity (BMI > 27.8 kg/m2), 27 with NGT and 17 with IGT, were recruited for the present study. Plasma IL-8 levels were measured in fasting state, after an oral glucose tolerance test (OGTT) and after euglycemic hyperinsulinemic clamp. Results The studied groups did not differ in fasting IL-8 concentrations. Both OGTT and clamp resulted in a significant increase in plasma IL-8. The change in IL-8 after clamp was similar in both groups. In contrast, after OGTT plasma IL-8 levels (IL-8OGTT) were markedly higher in IGT individuals. In IGT, but not NGT group, IL-8OGTT was positively related to postload glucose and negatively to insulin sensitivity. Conclusion Plasma IL-8 concentrations after glucose load are increased in obese IGT subjects in comparison to normoglycemic weight-matched individuals. Increase in plasma IL-8 might be both insulin-mediated (during clamp) and glucose-mediated (during OGTT).

It is well known that classification of diabetes in the group of patients aged 25-55 years, particularly not obese, is a difficult clinical problem. The aim of the study was to estimate clinical status of the investigated group and evaluate biochemical features based on positive titers of humoral immunological markers in the development of diabetes mellitus. We studied 31 non obese, newly diagnosed patients (mean BMI26 kg/m2), mean age--41.6 +/- 11.1 years, mean duration of diabetes mellitus was 11.4 +/- 21.8 months. Nearly 50% of the patients aged 25-55 years showed presence of humoral immunological markers, which suggests high prevalence of autoimmunological diabetes. These patients have a lower reserve of endogen insulin secretion. The evaluation at both B-cell secretory capacity and markers of autoimmunity is recommended to provide a proper classification.

Right classification of diabetes is important clinical issue. The aim of present study was to compare clinical, biochemical and immunological features, to analyze their practical use and to establish new decision tree which make the distinction between diabetes type 1, LADA, diabetes type 2 and MODY. We studied 97 not obese (mean BMI 26.3 +/- 4.9 kg/m2) patients aged 14 to 70 years, mean age 43 +/- 11.7 years, 53 women, 44 men. Mean duration of diabetes--2.3 +/- 4.3 years. We measured basal and stimulated C-peptide (6 minutes after 1 mg i.v. glucagon) (ELISA) and antibodies titers to glutamic acid decarboxylase--antiGAD65, tyrosine phosphatase-like molecule--IA2 and insulin--IAA (RIA). Autoimmune diabetes (LADA, type 1) was diagnosed with presence of one or more islet antigen antibodies. The highest frequencies had anti-GAD antibodies 33/97 (34%). The most complicated was to sort out group of patients with LADA. Comparison between this group and patients with diabetes type 2 have shown that BMI, co-existence of autoimmune disease, autoimmune markers and basal and stimulated C-peptide level measured at entry for the classification were useful in differentiation. Moreover we observed significantly lower C-peptide basal, stimulated and over basal level in group with MODY diabetes in comparison to diabetes type 2 patients. In the studied group were 5 patients with diabetes type 2 and obesity, in relatively young age. At the end there was one case of ADM (atypical diabetes mellitus). Clinical criteria for the classification of diabetes not always correlated with diagnosis. Autoimmune markers, basal and stimulated C-peptide were useful specially in differentiation between LADA and diabetes type 2 or diabetes type 1. Autoimmune diabetes co-existe with autoimmune disease. Proposed diagnostic scheme take for consideration presence of autoantibodies as well as C-peptide criteria.

Interleukin-18 (IL-18), a novel cytokine of the IL-1 family, and TGFbeta1 have been lately identified in several thyroid autoimmune diseases. The aim of this study was to estimate the influence of corticosteroids on serum IL-18 and TGFbeta1 in Graves' ophthalmopathy (GO) patients and to assess their potential as a guideline of immunosuppressive therapy. The study was carried out in three groups of subjects: (1). 17 patients with clinical symptoms of ophthalmopathy (GO) that underwent corticosteroid therapy consisting of intravenous methylprednisolone (MP) infusions and subsequent treatment with oral prednisone (P); (2). 14 patients with euthyroid Graves' disease (Gd) without symptoms of ophthalmopathy; (3). 12 healthy volunteers age- and sex-matched to groups 1 and 2. The serum levels of IL-18 and TGF beta1 were determined by the ELISA method. Thyroid receptor antibodies (TRAB) were estimated by the RIA method. Levels of IL-18 were significantly higher in both GO [340+/-109 pg/ml (p0.02)] and Gd individuals [308+/-89 pg/ml (p0.05)] as compared to the control group (238+/-88 pg/ml). There were no significant differences in TGFbeta1 concentrations between studied groups. TRAB values were significantly increased in both GO (p0.001) and Gd individuals (p0.001) as compared to the controls. In coticosteroid-responsive patients we have found a significant decrease in IL-18 serum concentration as compared to the pretreatment values. We did not find any correlation between IL-18 or TGFbeta1 and TRAB, duration of GO, clinical activity score (CAS) or proptosis. Our results suggest that efficient corticosteroid therapy in patients with Graves' ophthalmopathy may be related to its influence on systemic IL-18.

Platelet Na(+)/H(+) exchanger (NHE) activity, phospholipid-dependent thrombin generation, and platelet factor 3 (PF3) availability were measured in 83 type 2 diabetics and in 40 age- and sex-matched healthy subjects. Na(+)/H(+) exchanger activity was significantly increased in diabetic patients in comparison to the controls (kappa = 4.29 +/- 0.71 x 10(-3) x s(-1) v 3.21 +/- 0.64 x 10(-3) x s(-1), P.00001). However, there was no significant difference between subjects with (kappa = 4.28 +/- 0.75 x 10(-3) x s(-1)) and without (kappa = 4.26 +/- 0.32 x10(-3) x s(-1)) arterial hypertension, as well as between patients with normo- and microalbuminuria or overt proteinuria (kappa = 4.26 +/- 0.58 x 10(-3) x s(-1), kappa = 4.47 +/- 0.93 x 10(-3) x s(-1) and kappa = 4.07 +/- 0.38 x10(-3) x s(-1), respectively). Comparatively high NHE activity was observed in the group of patients with hemoglobin A(1c) (HbA(1c)) less than 7.5%. Multiple regression analysis revealed that the factors independently related to platelet Na(+)/H(+) exchanger activity were: total PF3 activity (beta = 0.77, P =.011) and triglyceride (TG) concentration (beta = 0.44, P =.039). Phospholipid-dependent thrombin generation and PF3 availability were also enhanced in all plasma fractions of diabetic patients, especially in platelet-poor plasma (PPP) and platelet-free plasma (PFP) (P.0001 and P.00001, respectively). There was a positive correlation between NHE activity and thrombin generation, as well as with PF3 availability in all plasma fractions. Our results suggest that enhanced platelet Na(+)/H(+) exchanger activity associated with raised phospholipid-dependent procoagulant activity may increase the risk of vascular damage in type 2 diabetic patients.

Hearing threshold of 40 patients with diabetes mellitus type 2 and 20 healthy controls were determined using pure tone audiometry of frequencies 500, 1000, 2000, 4000, 6000 and 8000 Hz and evoked otoacoustic emissions (EOAEs). The hypoacusis was bilateral, progressive and neurosensory nature, affecting the higher frequencies. Subjects with diabetes type 2 were more likely to have a hearing loss than were subjects without diabetes (95% vs 65%). This difference was statistically significant. Hearing loss was significantly correlated with age and duration of diabetes. Evoked otoacoustic emissions found out only in 2 patients with normal hearing. The results suggest that EOAEs doesn't show subclinical cochlear damage in diabetic patients because of the significant hearing loss.

IL-6 and its soluble receptor (IL-6R) appeared as reliable markers of inflammation activity in autoimmune diseases. The aim of the study was an estimation of serum IL-6 and sIL-R in patients with Graves' disease with ophthalmopathy during treatment with corticosteroids to assess their potential as a guideline of immunosuppressive therapy. We detected serum HIL-6 and IL-6R in three groups of subjects: 18 patients with clinical symptoms of ophthalmopathy (Clinical Activity Scoreor = 3, anamnesis of GOor = 1 yr), 16 patients with Graves' disease without ophthalmopathy (Gd) and 14 healthy volunteers. Corticosteroid therapy consisted of intravenous infusions of methylprednisolone (MP) and subsequent treatment with oral prednisone (P). The serum samples were collected 24 hours before MP, 24 hours after MP, 14 days of treatment with prednisone and after the end of the corticosteroid therapy. The levels of soluble IL-6 and IL-6R in the serum were determined by the ELISA method (Quantikine kit, RD Systems, Minneapolis). The statistical significance was estimated by the Mann-Whitney U-test. IL-6 concentration was significantly increased in patients with GO in comparison to the controls (12.4 +/- 3.7 vs 11.8 +/- 3.2; p0.05). After MP treatment in corticosteroid-responsive patients (improvement in CASor = 1) serum concentration of sIL-6R decreased significantly in comparison to pretreatment values (32.8 +/- 4.2 vs 28.6 +/- 4.9; p0.05). We found a positive correlation between IL-6 concentration and degree of proptosis.1. IL-6/IL-6R system plays an important role in the pathogenesis of GO. 2. IL-6R is a potent prognostic factor for efficacy of the immunotherapy but further investigations are needed.

Platelet sodium-proton exchange rate and phospholipid dependent procoagulant activity were measured in 31 type 1 diabetics (mean age 32.3 +/- 10.1 years) and 35 healthy subjects (mean age 35.4 +/- 9.4 years). The activity of platelet Na+/H+ exchanger was measured in platelet rich plasma, using an optical swelling assay, according to Rosskopf et al. Platelet procoagulant activity was measured in platelet rich plasma, platelet poor plasma and platelet/microparticles filtrated plasma, using Russell's viper venom (according to Jy and Horstman) and calibrated with ship L-alpha-phosphatidylethanolamine. We found that Na+/H+ exchange rate was significantly higher in diabetic patients in comparison to the controls (p = 0.0009). There was also a positive correlation between the activity of Na+/H+ exchanger and phospholipid dependent procoagulant activity in all plasma fractions. We did not find a significant association between Na+/H+ exchanger activity and metabolic parameters studied, however in patients with HbA1c level7.5% higher Na+/H+ exchange rates were noted. Total procoagulant activity did not rise significantly in diabetic patients, but was markedly higher in platelet poor and platelet filtrated plasma. It was supposed that it originated from platelet derived microparticles, enriched in phospholipids. Our results suggest that an increased platelet Na+/H+ exchange rate and raised procoagulant activity connected with platelet microparticles may enhance the risk of vascular damage in type 1 diabetic patients.

OBJECTIVE—Tumor necrosis factor-α (TNF-α) is one of the proposed mediators of insulin resistance, upregulated in human obesity. Insulin resistance, however, might precede the development of obesity, especially in subjects with a family history of type 2 diabetes. Therefore, the aim of the present study was to assess plasma levels of TNF-α and soluble forms of its receptors (soluble TNF-α receptors 1 [sTNFR1] and 2 [sTNFR2]) and to evaluate the relationship of the TNF-α system with insulin resistance in lean, nondiabetic offspring of type 2 diabetic patients. RESEARCH DESIGN AND METHODS—We compared 20 lean offspring (BMI RESULTS—Both men and women in the offspring group were markedly more insulin-resistant and had higher plasma levels of sTNFR2 (all P < 0.05). TNF-α, sTNFR1, and other examined parameters did not differ between the studied groups. Both TNF-α receptors were related to waist-to-hip ratio (WHR), fat-free mass (FFM), plasma total cholesterol, HDL cholesterol, LDL cholesterol, and nonesterified fatty acids (NEFAs). sTNFR2, but not sTNFR1, was also associated with insulin sensitivity (r = −0.49, P = 0.001). This relationship remained significant after adjustment for WHR, FFM, plasma insulin, and NEFA. CONCLUSIONS—TNF-α system might be involved in modulating insulin action before the onset of obesity in subjects at high risk for type 2 diabetes.