Polymorphism of catalase gene promoter in Romanian patients with diabetic kidney disease and type 1 diabetes.

Hyperglycaemia leads to ROS (Reactive oxygen species) generation, affecting the cells that cannot decrease glucose uptake such as: glomerular epithelial cells, mesangial cells and proximal tubule cells. ROS excess seems to activate important pathogenic pathways of development of diabetic nephropathy. The decrease of CAT activity, one of the most important antioxidant enzymes, following to some genetic defects, may be a risk factor for diabetic nephropathy. The purpose of this study is to investigate the association of 21A/T (rs7943316) polymorphism of CAT gene with advanced diabetic nephropathy in patients with type 1 diabetes in Romania. There have been studied 238 patients with T1D (type 1 diabetes), divided into the group with diabetic nephropathy (DN) (106 patients) and the group without renal affectation (132 patients). The genotyping has been made by using PCR-RFLP technique. The analysis of association has been made by using DeFinetti programme. The value considered significant has been p < 0.05. There has been a deviation from Hardy-Weinberg equilibrium in the group with diabetic nephropathy (p = 0.019), the equilibrium being preserved by the control group (p = 0.771). T allele does not confer a risk for advanced diabetic nephropathy (ORT = 0.757, 95% C.I. = 0.405-1.414; P = 0.381), the result being statistically insignificant even taking into consideration the risk allele A (ORA = 0.793, 95% C.I. = 0.465-1.350; P = 0.392). The results remain concordant too after applying the Cochran -Armitage test. Our data do not suggest an effect of 21A/T (rs7943316) polymorphism in the susceptibility for diabetic nephropathy in Romanian patients with type 1 diabetes. Further studies are necessary in order to demonstrate or exclude the role of CAT gene in diabetic nephropathy in patients with type 1 diabetes.

The effect of magnesium deficit on serum immunoglobulin concentrations in type 1 diabetes mellitus.

The aim of this study was to evaluate the correlation between changes in the concentration of serum magnesium and serum immunoglobulin concentrations in type 1 diabetes mellitus. In this study were included 110 patients with type 1 diabetes mellitus (64 men and 46 women) with ages ranging from 19 to 54 years (mean age 41.6+/-6.8 years). The mean duration of the disease was 8.7+/-7.5 years. Thirty-six healthy subjects served as a control group. The serum magnesium concentrations were evaluated by VITROS 750 XRC, Johnson & Johnson kit, (Ortho Clinical Diagnostics). Total serum IgA, IgG and IgM were determined by laser nephelometry (MININEPH The Binding Site kit). Values are means (x) + standard deviations (SD). Serum magnesium concentrations confirmed the magnesium deficit in patients with type 1 diabetes mellitus (1.8+/-0.11 mg/dL, range 1.73-2.47 mg/dL vs 2.2+/-0.2 mg/dL, range 1.6-2.4 mg/dL). In patients with type 1 diabetes mellitus, IgA levels are mildly elevated (4.03+/-0.51 g/L vs 3.43+/-0.48 g/L; p<0.05), while IgG levels are decreased (7.38+/-0.76 g/L vs 9.92+/-1.32 g/L; p<0.001) and IgM levels are almost constantly normal (1.18+/-0.16 g/L vs 1.22+/-0.15 g/L; p>0.05). Therefore, magnesium deficit has profound immunosuppressive capabilities in patients with type 1 diabetes mellitus by significantly reducing the number of IgG synthesizing cells and serum IgG concentrations.

The influence of sodium metavanadate on the process of diabetogenesis in BB rats.

Vanadium has been shown to be beneficial in the oral treatment of animal models of type 1 and type 2 diabetes. The aim of the study was to evaluate the short-term effects of sodium metavanadate in prediabetic BB-DP rats. To do this, 96 rats were divided into 4 equal groups. Groups V1, V2, V3 were treated with sodium metavanadate (0.1, 0.2 and 0.3 mg/ml respectively) and sodium chloride (0.5 mg/ml) in drinking water for 7 days. Group C received only sodium chloride (0.5 mg/ml). Blood glucose (BG), glycosuria, ketonuria, body weight and insulinemia were determined. The age of onset of diabetes was significantly higher for groups V2, V3 compared to group C, (p<0.05) and depends on the metavanadate concentration (V3 vs. V1, p=0.006). The incidence of diabetes was lower in the rats treated with metavanadate than in the control group, but this difference was not statistically significant. In diabetic rats, the BG at the onset was higher in group C than in groups V, p<0.05. Insulinemia, at the onset of the treatment as well as immediately after its cessation showed a drop in the treatment groups, proportionally to the dosage of vanadium, but later increased slowly and continuously until the end of the experiment. In conclusion, metavanadate delays the development of diabetes in BB-DP rats, but does not prevent its onset. A milder form of diabetes occurs in diabetic rats treated with metavanadate. The effects depend on the metavanadate concentration and 0.2 mg/ml is preferable.

Evaluation of the metabolic balance in type 2 diabetes by assay of the hair glucose.

The authors used a relative new method of long term retrospective evaluation of metabolic balance in type 2 diabetes. This method was based on the variations of the glucose in the hair with the variations of HbA1c (r=+0.96; p<0.001). In this way the authors could follow up the diabetic's metabolic balance on longer time periods than by determination of the HbA1 values.

The analysis of structural lipid of the hair of patients with angina pectoris.

A study of 48 patients with angina pectoris carried out, in which the structural lipids of the hair were analyzed and correlated with the manifestations of angina and the potential incidence of dyslipidemia. It was noted that the excesses of hair lipids associated with deficiencies of some unsaturated fatty acids raised the frequency of angina attacks in the group studied by 2.7 times. The normalization of the lipid levels diminished the levels of angina. Data were obtained on the influence over a long time (weeks, months) of the hypolipidemic treatment. The analysis of the hair has been shown to be also useful in detecting latent dyslipidemia associated with angina pectoris. Also, through the analysis of hair, large scale epidemiologic studies can be carried out on the association between dyslipidemia and angina pectoris.

The relationship between the lipid structure of the hair and acute myocardial infarction.

In a group of patients with acute myocardial infarction (AMI) an evaluation of the derangements in lipid metabolism was carried out by analysing the fat structure of the hair (closely reflecting serum lipid variations). The data obtained reflects the real incidence of dyslipidemia associated with myocardial infarction. Through hair analysis, the evaluation in time (over weeks, months) of lipids metabolism under hypolipidemic therapy can be carried out without taking repeated samples.

Animal models of type I (insulin-dependent) diabetes mellitus.

Much of our present knowledge concerning the etiopathogenesis, treatment and prevention of human diabetes would never have been acquired without the study of animal models of diabetes. The main models of IDDM may be divided into two groups: induced (through pancreatectomy, chemicals such as alloxan and streptozotocin, viruses and others) and spontaneous (mainly using BB rats and NOD mice). The latter, at different ages, develop a diabetic syndrome, with clinical characteristics, genetics and immunology that are very similar to the human disease. Among the more significant differences are lymphopenia (in BB rats) and the predominance of diabetes in females (in NOD mice). Studies aimed at preventing IDDM have advanced by leaps and bounds by using the two spontaneous models. These include various methods such as genomic modification, an influence over some environmental agents, immunosuppression, immunotherapy, immunomodulation and tolerance induction as well as protection of the beta-cell from autoimmune attack. The conclusions drawn from animal experiments have allowed some human trials to be carried out with encouraging results.

Safety and efficacy of insulin lispro in patients with diabetes mellitus.

Lispro is a human insulin analogue with a very rapid onset of action, and a shorter duration of activity than soluble insulin. In order to assess the therapeutical value of lispro, we have had an open-label, non-comparative study, for 12 weeks, involving 19 IDDM patients. The treatment regimen with lispro and Humulin N has been adapted depending on each patient characteristics. Patients attended three visits, and the main metabolic control parameters included values of hemoglobin Alc, fasting and postprandial blood glucose monitoring. The patients themselves monitored their blood glucose using a glucometer. The mean age value of 19 patients (8 females and 11 males) was 22.32 (+/- 13.59) years. In patients previously receiving insulin treatment, therapy with lispro insulin significantly reduced postprandial glucose values. Lispro has been administered t.i.d. in 14 patients, and b.i.d. in 5 patients. At visit 1, mean value of HbAlc was 10.32% (+/- 1.63%); at visit 3, mean HbAlc was 9.90% (+/- 1.59%). Total insulin daily dose and the rate of short and long acting insulin did not change from visit 1 to visit 3. There has been reported only one serious adverse event during the study: a ketoacidosis due to a technical dosing error. Ten patients have reported mild hypoglycemic episodes. The outcomes of clinical study and of Quality of Life Questionnaire suggests that lispro--the first human insulin analogue used in humans--is effective, safe, and it is broadening beneficially the spectrum of insulins.

Glimepiride-induced prevention of diabetes and autoimmune events in the BB rat: revised.

The purpose of this study was to determine whether Glimepiride, an oral sulfonylurea drug, prevents the onset of diabetes in diabetic prone BB rats. S750181, a sulfonylurea drug that has minimal in vivo glucose metabolic effects, was also tested. In addition, the shortest period of sulfonylurea treatment required for prevention was determined. Eighty rats were studied for all treatment periods with 40 receiving a daily oral gavage dosage of glimepiride and 40 receiving a daily oral gavage dosage of vehicle solution. Diabetes onset was monitored by glycosuria and blood glucose levels. In study I, with a treatment period of 35-142 days of age, Glimepiride-treated rats showed a 32% incidence of diabetes, whereas control rats had a diabetes incidence of 55% (p < 0.04). In study II, with a treatment period of 60-140 days of age, Glimepiride-treated rats showed a 29% incidence of diabetes compared to 54% in controls (p < 0.03). Further, comparing the time of diabetes onset between the Glimepiride and control groups showed that Glimepiride delays diabetes onset (p < 0.02). In study III, with a treatment period of 60-100 days of age, Glimepiride-treated rats showed a 17% overall diabetes incidence at 170 days, whereas the controls were 43% (p < 0.01). In study IV, with a treatment period of 60-140 days of age, S750181-treated rats showed a 38% diabetes incidence and the control group showed a 43% diabetes incidence. There was no significant delaying or prevention effect observed in the S750181 group. To determine if Glimepiride affected autoimmune events, the severity of islet inflammation was examined. In study I, islet histology from total and nondiabetic animals indicated that Glimepiride-treated rats had a lower severity of islet inflammation than that of the control rats (p = 0.023). These studies show that a) Glimepiride has diabetes preventive effects, b) shorter treatment periods of only 40 days can be effective and c) Glimepiride decreases the severity of islet inflammation.

The effects of nicotinamide and glimepiride on diabetes prevention in BB rats.

Glimepiride is an oral sulfonylurea drug; nicotinamide is an inhibitor of poly (ADP-ribose) synthetase and a precursor of NAD. Three studies were carried out to determine whether glimepiride and/or nicotinamide could prevent diabetes in BB rats. In Study I, we administered glimepiride treatment 200 mg/kg/day orally from the 35th to 143rd day of age. The incidence of diabetes in the glimepiride group was lower than in the control group (32% vs 55%, p < 0.02). In Study II, the treatment period was from the 35th to 147th day of age, and rats received glimepiride combined with nicotinamide (500 mg/kg/day IP). The treatment group showed a 22% incidence of diabetes compared to 53 in controls (p < 0.03). In Study III, nicotinamide treatment alone (1000 mg/kg/day orally) and combined with glimepiride were compared to untreated controls. The treatment period was from the 35th to 167th day of age. Nicotinamide-treated rats showed a 42% incidence of diabetes compared to 60% in controls (p = NS). In the nicotinamide combined with glimepiride treatment group, a lower incidence (28%) was observed when compared to the controls (60%, p < 0.05). These findings suggest that glimepiride can prevent diabetes in BB rats; however, nicotinamide when used in young animals shows only a trend to lower the incidence of diabetes, and when combined with glimepiride, no significant effect is observed.

Epidemiology of diabetes in Bucharest.

An analysis of the last 20,000 newly diagnosed diabetic patients consecutively registered from 1 January 1981 to 6 June 1991 in the Bucharest Registry of Diabetes showed the following: (1) primary insulin-dependence (Type 1 diabetes) was encountered in only 7% of cases: the rest were Type 2 diabetic patients (8745:43.7% treated with diet alone and 9856:49.3% treated with diet and oral drugs); (2) low body weight (BMI < 25) was encountered in 81.7% of patients in the age group 0-20 years, while obesity (BMI > 27) was encountered in 75.7% of cases in the age group 41-65 years; (3) the overall annual incidence of the Type 1 diabetes for all ages was 5.7/100,000, lowest (1.3/100,000) in the age group 0-4 years and the highest 10.1/100,000) in the age group 65-69 years; (4) the overall annual incidence for the Type 2 diabetes was 76.3/100,000, the lowest (2.4/100,000) in the age group 20-24 years and the highest (261.4/100,000) in the age group 60-64 years. Studying the relationship between the onset of Type 1 diabetes mellitus and age, we did not observe the previously reported strong relationship, so that the distribution of Type 1 diabetes can be considered relatively uniform, with the exception of extreme ages. In conclusion, each year, about 1 in 1000 inhabitants of Bucharest are registered as having diabetes, the majority (93%) had type 2 and only 7% had Type 1 diabetes, one of the lowest incidence rates in Europe.

Fear of hypoglycemia in type 1 (insulin-dependent) diabetic patients.

A clinical-psychological study was carried out in 224 insulin-dependent diabetic patients, distributed into two groups according to the frequency of hypoglycemic episodes. Group. A included 124 patients, 65 men and 59 women (mean age 35 +/- 18 years), who reported frequent episodes of hypoglycemia (HG). In the 100 patients of group B (46 men and 54 women with a mean age of 37 +/- 10 years), the HG episodes were only occasional. The degree and type of anxiety were evaluated using: (1) a semistructured interview to determine the HG frequency and intensity; (2) the anxiety scale for HG, with two sections: a) 15 items about the behaviour to prevent HG occurrence and b) 10 items for the anxiety level; (3) the questionnaire of anxiety with 40 items. Highly significant differences between groups were found for the degree of anxiety about HG, particularly for the frequency of high anxiety scores (with ideative ruminations or generalized anxiety, accompanied or not by agoraphobia) (73.86% in group A versus 34.09% in group B; p < 0.001). Relatively significant differences were recorded with respect to the compliance to treatment (preventive behaviour for HG) as well as for the anxiety index (47.95% in group A versus 33.45% in group B), with a prevalence of the covered anxiety in group A and of the overt anxiety scores, indicating a dissimilation tendency of the adaptative role of anxiety as a signal of communication.

The variations of the serum IgD and of the circulant immune complexes in diabetic patients.

The levels of serum IgD and of the circulating immune complexes (CIC) were determined in 168 diabetic patients, of whom 78 with type 1, 59 with type 2 and 31 with the so-called "intermediary" type of the disease, in comparison with 124 non-diabetic subjects for IgD and 100 for CIC. The results revealed very low IgD titres (less than 1 mg%, considered undetectable) in almost 3/4 of the cases; values over 1 mg% were recorded mostly in the cases of type 1, followed by those of "intermediary" and of type 2 diabetes. The mean CIC values of 67.13 +/- 36.53 optic density units (O.D.U.) were significantly higher than in the non-diabetic controls. Certain differences with respect to age, diabetes type, duration of the disease and of the insulin therapy were also recorded. The data are interpreted with caution, further investigations being necessary to the assertion of definite conclusions.

Not only quantitative but also qualitative changes of serum immunoglobulins in diabetes mellitus.

To extend previous observations on the quantitative changes of IgA and other serum Ig in diabetics, additional immunochemical investigations were carried out in 96 patients, 63 males and 33 females, mean age 43.5 +/- 15.7 years, 51 with type 1 (insulin-dependent) and 45 with type 2 (non-insulin-dependent) diabetes. The immunological data were correlated with the clinical-metabolic aspects. In the whole group, the IgA level was increased (144.1 +/- 57.2 I.U.). Significant differences were recorded with respect to age for IgG, to age and diabetes type for IgA, to sex for IgM. Qualitative Ig changes, reflecting disturbances of molecular structure, mainly for IgG, seldom for IgM, but never for IgA, were observed in 20% of the patients with both types of diabetes, more seldom in cases with long disease duration. The IgG with qualitative changes were purified and their functional capacity of inhibiting the natural cytotoxic activity (NK) was tested in comparison with that induced by pretreatment of the effectory cells with normal IgG. Some of these modified IgG showed a reduced capacity of inhibiting the NK activity. These data confirm the existence of certain quantitative changes of the main serum Ig in diabetics and reveal the presence of qualitative disorders of the IgG molecules, with consequences on their functionality.

No relationship between insulin antibodies and hypoglycemia in insulin-treated diabetic patients.

It has been speculated that insulin antibodies may contribute to the hypoglycemic attacks in insulin-treated diabetic patients. To address this hypothesis, we analyzed in a first part of the study the frequency to hypoglycemia in two groups of diabetic patients, one (Group A, 38 cases) with at least two episodes of severe hypoglycemia in the last year and another (Group B, 38 cases) without severe hypoglycemia in the last 3 years. In the second part of this study, we analyzed the frequency of severe and moderate episodes of hypoglycemia in another two groups of diabetics, one (Group C, 32 cases) with high insulin antibody titre (greater than or equal to 20% binding, mean +/- SD 31.2 +/- 8.1%) and another with low insulin antibody titre (less than 10% binding, mean +/- SD, 5.1 +/- 2.2%). No significant difference was found for bound insulin between diabetics with frequent hypoglycemic episodes (2.3 +/- 0.2/patient/year--Group A) and those without severe hypoglycemic episodes (Group B), i.e., bound insulin 4.89 +/- 3.21% in group A versus 5.32 +/- 4.5% in group B. Conversely, the frequency of severe episodes of hypoglycemia was similar in diabetic patients with high (31.2 +/- 8% binding in group C) and respectively low (5.1 +/- 2.1% binding in group D) insulin antibody titre, i.e., 0.15 episodes/patient/year in group C and 0.17 episodes/patient/year in group D.

Immunological aspects of diabetes.

Notable progress was achieved by the investigations on the immunology of diabetes. Studies of immunogenetics have demonstrated that Type 1 (insulin-dependent) diabetes shows a primary association with the HLA class II genes (e.g., with HLA-DR3 and DR4 in Caucasoid populations), a determining features of this type of the disease. Besides, it presents a wide variety of autoantibodies, such as islet-cell cytoplasmic antibodies (ICA), islet-cell surface antibodies (ICSA), complement-fixing islet-cell antibodies (CF-ICA), antibodies to a Mr-64000 islet-cell protein, and many studies are trying to evaluate their pathogenic and predictive role. Various changes of cell-mediated immunity have been also described. The proportion of activated T cells in circulation is increased in patients with Type 1 diabetes. It was assumed that the T cells are the main cause of pancreatic beta-cell damage. Type 1 diabetes is actually considered as a chronic autoimmune disease, with several stages of evolution leading to the destruction of the pancreatic beta-cells, with a consecutive gradual decrease of insulin secretion. From the therapeutic point of view, many problems are raised by insulin immunogenicity, mainly depending on the so-called "contaminants". To avoid phenomena of allergic reactions, immunological insulin-resistance, lipodistrophy, a.o., highly purified and human type insulins have been prepared. The insulin autoantibodies (IAA) detected before the clinical onset of Type 1 diabetes are considered a new marker of autoimmunity. New hopeful prospects are opened by diabetes immunotherapy, in which Cyclosporin A detains a particular role, although it should be used only in special conditions, under strict clinical observation.(ABSTRACT TRUNCATED AT 250 WORDS)

A study on the types of diabetes mellitus in first degree relatives of diabetic patients.

The genetic characteristics of the diabetic types have been assessed by following up their frequency in first degree relatives of some non-selected diabetic patients, registered at eight different centers of the country. Out of 1,003 non-diabetic controls only 46 (4.6%) had 52 diabetic relatives, 65.4% of type 2 (non-insulin-dependent). Comparatively, out of the 704 patients, 172 (24.4%) had 229 diabetic first degree relatives, 72.5 of type 2. Out of 231 type 1 (insulin-dependent) diabetic patients, 29 (12.6%) had 34 diabetic relatives, 55.9% of type 1. Out of 300 type 2 patients, 99 (33.0%) had 121 diabetic relatives, 84.0% of type 2. The other 173 diabetic patients presented an "intermediary" type of the disease (needing insulin many years after onset). Forty-four (25.4%) of them had 64 diabetic relatives, 67.2% of type 2, 20.3% of type 1 and 12.5% with "intermediary" diabetes. The five times higher frequency of diabetes in patients' relatives versus controls is pointed out. Type 2 diabetic relatives predominated. The proportion of probands with diabetic relatives increased from 4.6% in non-diabetics to 12.6% in type 1, to 25.4% in "intermediary" diabetes and to 33.0% in type 2. The heredity of type 1 prevailed in type 1 and that of type 2 in type 2 and in "intermediary" diabetes. The fact that "intermediary" diabetes tends towards type 1 (insulin-dependent) as therapy and towards type 2 (non-insulin-dependent) as heredity might be an argument supporting the controversy on the diabetic syndrome classification.

Comparative study of insulin antibodies in diabetic patients with primary insulin-dependence (type I) and secondary insulin-dependence (type II).

Insulin antibodies (% binding) were determined by RIA method in 404 insulin-treated diabetic patients divided into two groups: (A) primary insulin-dependent patients (Type I diabetes): 300 cases, 170 M, 130 F, mean age +/- SD 29.2 +/- 7.5 yrs, disease and insulin treatment duration 7.7 +/- 6 yrs: (B) Type II diabetic patients needing insulin (secondary insulin-dependence): 104 cases, 47 M, 57 F, aged 53.4 +/- 9.2 yrs, duration of diabetes 13.1 +/- 8.3 yrs, and of insulin treatment 3.1 +/- 2.1 yrs. Both groups of patients were with the same types of insulin preparations. In 297 cases, all belonging to group (A), fasting C-peptide was also determined. The titre of insulin antibodies was significantly (p less than 0.001) higher in patients with secondary insulin dependence than in those with primary insulin dependence (22.96 +/- 15.1% vs 10.25 +/- 9.89) in spite of the longer duration of insulin treatment in the later group; the mean C-peptide value found in 58 Type I diabetic patients with a binding capacity less than 10% was significantly lower (p less than 0.001) than that found in 11 Type I diabetic cases with a binding capacity greater than 20% (0.091 +/- 0.57 vs 0.273 +/- 0.37 pmol/ml); no correlation was found between insulin antibodies and metabolic control, insulin requirements or chronic complications.