Małgorzata Szelachowska

Plasma interleukin 8 concentrations in obese subjects with impaired glucose tolerance.

BackgroundInterleukin 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).MethodsA 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.ResultsThe 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.ConclusionPlasma 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).

Effect of glycosaminoglycans on urinary albumin excretion in insulin-dependent diabetic patients with micro- or macroalbuminuria

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.

CCN2 protein is an announcing marker for cardiac remodeling following STZ-induced moderate hyperglycemia in mice

Diabetes causes changes in the myocardium, which are often called diabetic cardiomyopathy. This condition has been extensively investigated in animal models with high glucose levels. Nevertheless, it has not been investigated whether moderate hyperglycemia, in the absence of other features of metabolic syndrome, may also cause similar changes in the heart. The aim of the study was to assess changes in the myocardium in an animal model of mild type 1 diabetes. Moderate hyperglycemia was induced in 8- to 10-week-old male C57BL6J mice by 5 intraperitoneal injections of streptozotocin (40 mg/kg). After 16 weeks, they were sacrificed, and left ventricle (LV) dimensions and extent of cardiac fibrosis were assessed by morphometry. The abundance of CCN proteins in LVsamples was assessed using western blotting, while activity of metalloproteinase 2 was established in zymography. Real time PCR was used to investigate the expression of transforming growth factor beta1 (TGFbeta1) and atrial natriuretic peptide. Mice with moderate hyperglycemia presented comparable cardiac dimensions with fibrosis and hypertrophy parameters as the non-diabetic controls. However, the abundance of profibrotic CCN2 protein was significantly increased in hyperglycemic animals (1.67 +/- 0.28 vs. 1 +/- 0.47, p < 0.05). Interestingly, this change was independent from the TGFbeta1 expression, as its RNA abundance was similar in both groups. Moderate hyperglycemia also caused an increase in the activity of the metalloproteinase 2 (1.21 +/- 0.17 vs. 1 +/- 0.07, p < 0.05). Despite diabetes, no profound changes in cardiac morphology were found. In our animal model, moderate hyperglycemia caused activation of a profibrotic gene expression program, which was counterbalanced by the increase of metalloproteinase activity.

Nicotinamide inhibits enhanced in vitro production of interleukin-12 and tumour necrosis factor-α in peripheral whole blood of people at high risk of developing Type 1 diabetes and people with newly diagnosed Type 1 diabetes

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.

Association between type 1 diabetes and periodontal health

PURPOSE We assessed periodontal status in patients with type 1 diabetes and healthy individuals in relation to their glycemic control, smoking and inflammatory biomarkers. MATERIAL/METHODS Periodontal status was examined in 107 patients with diabetes and 40 controls, using Oral Hygiene Index (OHI), Community Periodontal Index (CPI) and tooth number. CPI values of 0-2 and 3-4 were classified as non-periodontitis and periodontitis, respectively. Blood samples were analyzed for glucose, HbA1c, CRP, fibrinogen, interleukin-1 and tumor necrosis factor-alpha (TNF-α). RESULTS Periodontitis was found in 15.0% of the controls and 57.9% of diabetic patients, including 40.0% of these with good metabolic control (GMC) and 59.5% of those with poor metabolic control (PMC). Severe periodontitis was more frequent in the PMC than in the GMC group and in the controls (26.0% vs. 20.0% vs. 5.0%). The PMC patients had lower number of sextants with CPI 0 and higher number of sextants with CPI 3 and CPI 4 as well as lower tooth number in comparison with the controls. The patients with periodontitis had higher TNF-α (p<0.001) and OHI (p<0.001) than the patients without periodontitis. The number of sextants with CPI 0 correlated negatively with fibrinogen and TNF-α levels, whereas the number of sextants with CPI 3 correlated positively with TNF-α and fasting glucose level. CONCLUSIONS There is good evidence that type 1 diabetes increases the risk of periodontal disease. Our results suggest that poor metabolic control of diabetes together with smoking and inadequate oral hygiene increase the risk of severe periodontal destruction in patients with type 1 diabetes.

Glycosaminoglycans urinary excretion as a marker of the early stages of diabetic nephropathy and the disease progression

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.

HLA-DR, HLA-DQB1 and PTPN22 gene polymorphism: association with age at onset for autoimmune diabetes.

Introduction Autoimmune diabetes has different clinical manifestations related to the age at onset. It is divided into several subtypes, including “classical” type 1 diabetes (T1D) and latent autoimmune diabetes in adults (LADA). The LADA is considered a slowly progressing subtype of autoimmune diabetes, although the clinical picture is more similar to type 2 diabetes. Material and methods The aim of this study is to investigate whether genetic predisposition influences age at onset in autoimmune diabetes. We studied rs2476601 PTPN22 gene polymorphism and HLA DR, HLA-DQB1 in 175 patients with classical type 1 diabetes, 80 LADA, and 151 control subjects from north-east Poland. Results The frequencies of the PTPN22 TT genotype were higher in the group of patients with classical type 1 diabetes (6.3%) and LADA (11.3%) than in control subjects (0.7%) (p = 0.02 and p = 0007, respectively). In patients with classical type 1 diabetes we observed an increasing trend in frequencies of genotype TT dependent on age at onset (3.9% (0-5 year olds), 6.0% (6-15 year-olds), 8.2% (16-25 year olds), p = 0.048). The incidence of predisposing human leukocyte antigen (HLA) genotypes HLA DR3/DQB1*02 and DR4/DQB1*0302 was found to decrease in the group with type 1 diabetes in relation to age at onset and LADA (HLA DR3/DQB1*02 – 69.2% (0-5 year olds), 57.0% (6-15 year olds), 51.0% (16-25 year olds), 46.3% (LADA), p = 0.032; HLA DR4/DQB1*0302 – 80.8% (0-5 year olds), 63.0% (6-15 year olds), 51.0% (16-25 year olds), 43.8% (LADA), p = 0.0003), and to increase for the protective allele DQB1*0602 (0.0% (0-5 year olds), 1.0% (6-15 year olds), 2.0% (16-25 year olds), 6.3% (LADA), p = 0.029). Conclusions Thus, age at onset for autoimmune diabetes appears to be related to a combination of predisposing and protective HLA alleles. Against a background of HLA genetic predisposition, other non-HLA loci may influence age at onset for late autoimmune diabetes.

The Changes in the Endothelial Function and Haemostatic and Inflammatory Parameters in Subclinical and Overt Hyperthyroidism

Introduction. The aim of the present study was to compare the levels of circulating markers of endothelial function and low-grade inflammation in patients with subclinical and overt hyperthyroidism (OH) due to Graves disease (GD) and toxic nodular goiter (TNG). Material and Methods. The group studied consisted of 42 patients with GD, 75 patients with TNG, and 39 healthy controls. Results. Circulating markers of endothelial dysfunction were elevated in the patients with both SH and OH, but the concentrations of interleukin-12 (IL-12) (P < 0.05), IL-18 (P < 0.05), fibrinogen (P < 0.01), and von Willebrand factor (vWF) (P < 0.05) were significantly higher in the OH than in the SH group. The highest levels of IL-6, IL-12, IL-18, vWF, sVCAM-1, and fibrinogen were found in the patients with GD, but the differences between the GD, and TNG groups were not significant. In the subjects with OH serum IL-6 was positively associated with FT3 (R = 0.276, P < 0.05), FT4 (R = 0.273, P < 0.05), and thyroid peroxidase antibodies (R = 0.346, P < 0.01) levels. Conclusion. Our results may suggest that both SH and OH may be associated with endothelial dysfunction, which is reflected by decreased fibrinolytic activity, hypercoagulability, and increased levels of IL-6, IL-12, and IL-18 and depends not only on the cause but also on the degree of hyperthyroidism.

Latent autoimmune diabetes in adults in a population-based cohort of Polish patients with newly diagnosed diabetes mellitus

Introduction Latent autoimmune diabetes in adults (LADA) is a slowly developing form of autoimmune diabetes characterized by the presence of type 1 diabetes-associated autoantibody. The aim of this study was to determine the incidence and characteristics of LADA in a population-based cohort of Polish patients with newly-diagnosed diabetes. Material and methods The study cohort was taken from the resident population of the city Białystok, Poland, during the period 1 January to 31 December 2003, aged 20-64 years. During this period we identified 231 cases of diabetes. We measured glutamic acid decarboxylase (GADA) and insulin antibody (IAA), insulin, C peptide and glycated hemoglobin (HbA1c). Diagnosis of LADA was made according to Immunology Diabetes Society and Action LADA criteria. Results The incidence of LADA was 10 per year per 100 000 people. The proportion of patients with LADA was 8.9% among newly diagnosed cases with diabetes. Patients with LADA were younger at diagnosis (48.5 ±9.4 years vs. 54.8 ±10.6 years, p < 0.01), had lower body mass index (26.9 ±9.3 kg/m2 vs. 29.5 ±5.2 kg/m2, p < 0.05), C peptide (126 ±127 pmol/l vs. 446 ±592 pmol/l, p < 0.001), and were less insulin resistant (HOMA IR 0.94 ±0.85 vs. 3.6±4.4, p < 0.001) compared to patients with type 2 diabetes. Glycated hemoglobin and fasting glucose were similar in patients with LADA and type 2 diabetes. Conclusions In addition to GAD, anti-insulin antibodies are useful for diagnosing autoimmune diabetes in adults. Patients with LADA have similar glucose control parameters (HbA1c) compared to patients with type 2 diabetes, although they are usually younger and have a lower body mass index. Patients with LADA make up a significant proportion of newly diagnosed people with diabetes mellitus in a Polish population.

γδ T-cells alterations in the peripheral blood of high risk diabetes type 1 subjects with subclinical pancreatic B-cells impairment

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.