[Frequency view on genome changes testing]. / Frekvencní pohled na vysetrení odchylek genomu.

Laboratories dealing with human genome, both inherited and acquired changes, dispose with similar methods and technology. The spectrum of genetic tests is relatively broad and the number of mutations or variants tested differs substantially. Also the number of examinations carried out in individual laboratories varies. Data presented in the tables come from the year 2004 and indicate the number of examinations requested and number of positive results. Many laboratories mentioned in the registry CZDDNAL (http://www.uhkt.cz/lab_a_vysetreni/nr lab_dna_diag/dna_lab_db) perform the same tests but there is also a great number of tests carried out by only one laboratory. Reasons of the request, cost-effectiveness and clinical utility of genetic testing is being discussed.

Insulin resistance and compensatory insulin secretion in middle-aged persons with hypertriglyceridemia.

Insulin secretion and insulin sensitivity were measured in hypertriglyceridemic patients using the frequently sampled intravenous glucose tolerance test. Three groups of men who were matched for age and body mass index were studied: eight healthy control subjects (C), seven patients with mild hypertriglyceridemia and normal glucose tolerance (TG), and seven with well-controlled type 2 diabetes with hypertriglyceridemia (TG-DM). The first-phase insulin response was increased by 116% in the TG group and decreased by 59% in the TG-DM group. The second phase of insulin secretion was increased in both TG groups (TG by 310% and TG-DM by 250%). The mean insulin sensitivity index (SI) was reduced by 50% in the TG group and by 60% in the TG-DM group. Glucose effectiveness (SG) was reduced by 30% in the TG-DM group compared with the control subjects.

Adrenergic overactivity and insulin resistance in nonobese hereditary hypertriglyceridemic rats.

Nonobese, hereditary hypertriglyceridemic (HTG) rats provide an interesting model of hypertriglyceridemia, glucose intolerance, and hypertension. In age-matched 15 HTG and 16 control Wistar rats fed on a high sucrose diet (70 cal%) for 6 weeks, we measured insulin sensitivity in vivo and some parameters of sympatoadrenal system. Using euglycemic clamps with administration of 2-deoxy[1-3H]glucose, we found whole body insulin resistance and decreased glucose metabolic index Rg' in soleus muscle, epitrochlearis muscle, diaphragm, and white adipose tissue in HTG rats. We found higher levels of plasma epinephrine and higher excretion of vanilmandelic and homovanilic acids in HTG rats. The binding of [3H]-dihydroalprenol to the heart membrane fraction was similar in both groups, but the dissociation constant Kd was increased by 75% in the heart of HTG rats.

Muscle GLUT 4 protein levels and impaired triglyceride metabolism in streptozotocin diabetic rats. Effect of a high sucrose diet and fish oil supplementation.

Our data indicate that (a) the existence of a defect in the clearance of circulating TG and persistence of muscle TG deposition in the high sucrose-fed neonatal streptozotocin diabetic rat, which (b) can only be partially corrected by raised dietary n-3 PUFA intake. (c) Skeletal muscle of STZ type II-like diabetic rats contains about 40% less glucose transporters, and (d) this situation cannot be changed by any of the dietary treatments employed. (e) These findings indicate that muscle TG accumulation may have no direct relation to glucose transport in muscle.

Comparison of N-acetyl-beta-glucosaminidase and albuminuria with clinical finding of microangiopathy in type I diabetes mellitus.

N-acetyl-beta-Glucosaminidase activity in serum and urine and microalbuminuria were measured in 70 type-I diabetics and compared with glycated serum protein as well as with the finding of diabetic retinopathy. A significantly increased N-acetyl-beta-glucosaminidase activity in serum correlated positively with glycated protein but not with the development of retinopathy. Urinary N-acetyl-beta-glucosaminidase activity and albuminuria were significantly increased in diabetics with (p less than 0.001) or without (p less than 0.01) retinopathy as compared to healthy controls. A significant positive correlation was observed between urinary N-acetyl-beta-glucosaminidase activity and albuminuria (r = 0.73, p less than 0.01) as well as between blood pressure and albuminuria (r = 0.51, p less than 0.05).

N-acetyl-beta-glucosaminidase and albuminuria in normal and diabetic pregnancies.

Fourteen diabetic women without signs of nephropathy were examined during pregnancy. Serum fructosamine concentration indicating short-term metabolic control of diabetes was normalized at the beginning of the second trimester and was within the normal limits till the delivery. A gradual increase of N-acetyl-beta-glucosaminidase activity in serum and urine has been found during pregnancy in diabetic and healthy women. No significant differences of N-acetyl-beta-glucosaminidase activities were observed between the above groups. A successive increase of albuminuria during pregnancy was present in diabetic and healthy women with about 10-times higher values at delivery. A significant positive correlation was observed between albuminuria and urinary NAG activity in both groups of pregnant women (r = 0.77). We did not find any deterioration in N-acetyl-beta-glucosaminidase activities and albuminuria in seven diabetic women one year after delivery.

Early changes of serum N-acetyl-beta-glucosaminidase, tissue plasminogen activator and erythrocyte superoxide dismutase in relation to retinopathy in type 1 diabetes mellitus.

Biochemical markers of early changes that are characteristic for diabetic microangiopathy are not completely understood. We investigated activities of serum N-acetyl-beta-glucosaminidase (NAG), tissue plasminogen activator and erythrocyte superoxide dismutase in well defined groups of type 1 diabetic patients. Patients were selected on the basis of 4 year follow-up observation. Forty-two type 1 diabetic patients were subdivided into those without retinopathy (n = 13) throughout the study, those with newly developed or worsened retinopathy (n = 12) during 4 years and those with retinopathy already established at the beginning of the study and without evidence of its progression (n = 17). All diabetic patients had albustix-negative urine. A significant increase of the mean serum NAG activity during 4 years was found only in patients without retinopathy (P < 0.01) whereas no changes of the altered enzyme activities were present in patients with developing and established retinopathy. The mean activity of tissue plasminogen activator was elevated in all groups of diabetic patients compared with healthy subjects (P < 0.001). A significant positive correlation was found between plasminogen activator and serum NAG (r = 0.51, P < 0.01). Erythrocyte superoxide dismutase was higher in diabetic patients than in healthy persons (P < 0.01) but no differences were observed between the patients with or without retinopathy. Superoxide dismutase positively correlated with NAG (r = 0.57, P < 0.01). We conclude that early functional changes precede a morphological development of diabetic retinopathy as was evident from the altered enzyme activities.

The hypertriglyceridemic rat as a genetic model of hypertension and diabetes.

Hypertriglyceridemia was demonstrated in untreated hypertensive patients as well as in animals with genetic and experimental hypertension. The main purpose of the present study was to evaluate the possibility to use the hereditary hypertriglyceridemic (HTG) nonobese rats in hypertensive research. Direct measurement of blood pressure demonstrated significantly higher systolic, diastolic and mean arterial pressures in HTG rats in comparison with control Wistar rats. There was significant positive correlation between blood pressure and plasma triglyceride concentration (r = 0.585, n = 40, p less than 0.001). In addition, there were significantly increased plasma norepinephrine and epinephrine concentrations in HTG rats, suggesting that the stimulation of sympathetic nervous system could be one of the pathogenetic mechanisms involved in the increase of blood pressure of HTG rats.

Epidermal growth factor (EGF) in serum of patients with differentiated carcinoma of thyroids.

Serum levels of epidermal growth factor (EGF) were investigated in 31 patients with differentiated carcinoma of thyroid. Patients with carcinoma had significantly decreased basal levels of serum EGF, however this decrease in serum EGF occurred only in the group of patients which had been evaluated at an interval of six weeks after ablation of residues of normal thyroid with radioiodine. In patients that had been treated with radioiodine for cancer this decrease was not observed. The change in serum EGF was not dependent on thyroidal function and did not correlate with the serum thyreoglobulin level. It is presumed that decreased serum EGF is a consequence of functional changes of platelets after whole body irradiation at ablative dose of radioiodine.

Blood pressure changes induced by chronic insulin treatment in Wistar rats.

Hyperinsulinaemia may play a causal role in the development of hypertension in obese hypertensives. However, experimental evidence supporting this statement is inappropriate. The main purpose of this study was to evaluate the chronic effects of insulin administration on blood pressure, total-body glucose metabolism and urinary catecholamine excretion. After 10 weeks of insulin injection blood pressure was substantially increased in insulin-treated animals compared to those treated with saline (125 +/- 2 vs 108 +/- 2 mm Hg, p < 0.001). There were no differences in glycaemia, plasma triglyceride levels and free fatty acid levels between these two groups. Plasma level of corticosterone was increased in both insulin-treated and saline-treated rats as compared to untreated animals suggesting that the level of stress was similar in both injected groups. The urinary excretion of norepinephrine and dopamine was increased in the insulin-injected group by about 120% and 310%, respectively. Our data clearly indicate that long-term insulin administration increased blood pressure but the underlying mechanisms remain to be elucidated.

Estradiol-induced adenohypophyseal growth reaction and beta-adrenergic receptors.

The effect of administration of estradiol benzoate on beta-adrenergic receptors of rat adenohypophyseal cells was studied. Twenty days' administration of estradiol benzoate was followed by an increase of adenohypophyseal weight and a decrease in specific binding of 3H-dihydroalprenolol (3H-DHA). In contrast to thyroid hormone treatment which induced an increase in 3H-DHA binding, thyroid hormone treatment decreased both the growth reaction and the reaction of beta-adrenergic receptors after estradiol. Although the relationship between the adenohypophyseal receptors and the growth reaction is unclear, changes in beta-adrenergic receptors after hormonal therapy can be one of pathophysiological conditions that may influence this reaction.

A monoclonal antibody applicable for determination of C-peptide of human proinsulin by RIA.

BALB/c, (BALB/c x B10.A)F1 and (BALB/c x B10)F1 hybrid mice were immunized with C-peptide of human proinsulin. The (BALB/c x B10.A)F1 hybrids were the best responders and yielded 3 hybridomas secreting specific monoclonal antibodies. One of them, C-PEP-01, bound the C-peptide with high affinity (Kas = 1.1 x 10(9) l/mol), cross-reacted fully with human proinsulin but not with insulin, glucagon or somatostatin and apparently recognized the regions of C-peptide comprising amino acid residues 8-13 and 25-31. A RIA system could be set up employing this monoclonal antibody suitable for estimation of C-peptide concentrations in a diagnostically useful range (1-50 ng/ml).

[Neuroendocrine carcinoma of the pancreas with the hyperinsulinism syndrome]. / Neuroendokrinní karcinom pankreatu se syndromem hyperinzulinismu.

In a 26-year-old woman with symptoms of hyperinsulinism explorative laparotomy revealed a pancreatic tumour metastatizing into the liver. Intensive cytostatic therapy led to a temporary inhibition of hyperinsulinism, however, in the course of two years massive infiltration of the retroperitoneum, left adrenal, gastric wall, liver, mesentery and abdominal lymph nodes by a tumour occurred. On necroptic examination the tumour had characteristics of a neuroendocrine carcinoma with carcinoid features. Part of the tumour cells were argyrophil; reliable evidence of insulin production, which during the terminal stage of the disease played again a major part in the clinical picture, was made possible only by the use of a very sensitive Czech made antibody against C peptide.

[Endothelial dysfunction in diabetes mellitus]. / Endoteliální dysfunkce u diabetes mellitus.

Impairment of the vascular endothelium and its functions is a common sign of several serious diseases (atherosclerosis, hypertension, vascular spasms, thromboses) and is the initial stage of vascular affection in diabetes mellitus. The endothelium plays an important role in the transformation of some substances with a cardiovascular action and it secretes itself vasoactive substances. Vascular affections in diabetes are characterized by impaired homeostasis of vasoactive substances of endothelial origin--raised levels of vasoconstrictor factors (endothelins, thromboxanes) and reduction of vasodilatating factors (prostacyclin, EDRF--endothelin derived relaxing factor) as well as disorders of their interrelations. Vasoactive agents lead at the same time also to alteration of the growth and proliferation potential of smooth muscle cells of the vascular wall and thus to remodelling of the vascular structure in diabetes. At present possible ways how to influence these processes in a favourable way are intensely studied and discussed.

[Insulitis--new findings on the role of genetic and external factors]. / Insulitis--nekteré nové názory na roli genetických a zevnich faktoru.

The autoimmune process focused on B-cells of the islets of Langerhans and leading gradually to their destruction and the development of type I diabetes takes place under the morphological picture of insulitis. The disease affects genetically predisposed individuals. At the end of the eighties knowledge was greatly expanded due to methods of molecular genetics. The majority of authors assumes that in the pathogenesis of insulitis also environmental factors participate. More detailed information is provided on chemical compounds, nutritive substances and viruses.

[Insulin secretion and sensitivity in middle-aged men in the initial stages of type II diabetes mellitus (evaluated by the "minimal model" method]. / Inzulínová sekrece a senzitivita u muzu stredního veku v iniciálním stadiu diabetes mellitus II. typu (hodnocená metodou "minimálního modelu").

The objective of the work was to test the minimal model method based on computer evaluation of parameters (blood sugar level, insulin blood level) obtained during the intravenous glucose tolerance test with frequent collection of samples, when examining the insulin glucose homeostasis in patients in the initial stage of type II diabetes. The authors examined a control group of 8 healthy men, mean age 43.6 years, BMI 24.7 and a group of 7 men with type II diabetes, mean age 46.7 years, BMI 31.37. Indexes (phi 1 and phi 2--the first and second stage of insulin secretion, SG-glucose efficiency, SI-insulin sensitivity) were obtained by evaluation of the results, using a programme for a personal computer PC AT. In men of the control group the authors recorded the following values: phi 1 5.80 +/- 1.26 microU/ml x min/mg/dl, phi 2 6.43 +/- 5.15 microU/ml x min-2/mg/dl, SG 0.016 +/- 0.010 min-2. Diabetic patients were divided with a regard to the first stage of insulin secretion into two groups: the first one (n = 4) comprises those where the first stage is minimal (phi 1 = 0.80 +/- 0.68 microU/ml x min/mg/dl, phi 2 = 33.41 +/- 19.06 microU/ml x min-2/mg/dl, in the second group are those (n = 3) where the first stage of insulin secretion is maintained phi (1 12.23 +/- 5.51 microU/ml x min-2/mg/dl, phi 2 7.77 +/- 4.98 microU/ml x min-2/mg/dl). The second stage of insulin secretion in subjects of the first group is, as compared with controls, significantly higher (p < 0.05).

[Hyperinsulinemia--the common denominator in type II diabetes mellitus,obesity, hypertension, hypertriglyceridemia and atherosclerosis]. / Hyperinzulinémie--spolecný jmenovatel pro diabetes mellitus II. typu, obezitu, hypertenzi, hypertriglyceridémii a aterosklerózu.

During the last twenty years we witnessed a remarkable increase in knowledge of the mechanism as regards insulin action, the central hormone of metabolic regulations. Interest in cellular and molecular mechanisms of action was conditioned by a high prevalence of insulin resistance and the fact that insulin resistance holds a key position in the pathogenesis of many diseases, in particular atherosclerosis, obesity, hypertension, diabetes mellitus type II, ovarian hyperandrogenism and others. The syndrome of hyperinsulinaemia/insulin resistance is the basic component of the so-called X syndrome defined in 1988 by Reaven. It is encountered in subjects with a normal glucose tolerance but a predisposition for diabetes type II. If this disposition, probably genetic by nature, is potentiated by the central type of obesity and a sedentary lifestyle it can influence the development of hypertension and dyslipidemia. The sum of these factors promotes acceleration of atherosclerosis and frequently its premature manifestations: myocardial infarction and other cardiovascular diseases which hold the first place as regards causes of death on a world wide scale. It is important to identify but also to treat this complex not only metabolic risk factors for macrovascular diseases. It is a paradox that some drugs used as antihypertensives can cause deterioration of insulin resistance, subsequently influence in an adverse manner dyslipidemia and thus increase the metabolic risk of cardiovascular diseases. In the submitted paper the authors tried to summarize hitherto expressed views on the syndrome of hyperinsulinaemia and insulin resistance, using as a basic the results of their own work.

[The effect of fish oil on metabolic parameters in patients with type 2 diabetes mellitus associated with dyslipidemia]. / Ucinky rybích oleju na nekteré metabolické parametry nemocných s diabetes mellitus 2. typu a pruvodnou dyslipidémií.

BACKGROUND: The authors discuss the effect of administration of fish oils rich in n-3 fatty acids in diabetic patients type 2 and draw attention to the possible deterioration of glucose homeostasis. The objective of the investigation was to assess changes of the lipid and carbohydrate metabolism in type 2 diabetics with associated dyslipidaemia after enrichment of the diet with n-3 fatty acids. METHODS AND RESULTS: To 17 patients with type 2 diabetes and dyslipidaemia fish oil containing 5.5 g n-3 fatty acids per day was administered. After six weeks a decline of triglycerides was recorded (-47%, P < 0.01), free fatty acids (-27%, P < 0.01) and a rise of HDL2-cholesterol (25%, P < 0.05). The concentration of apo B, apo A-1, LDL- and HDL cholesterol did not change significantly. There were no significant changes of the blood sugar level, glycosylated haemoglobin and fructosamine. The insulin and C-peptide concentration on fasting (and after glucagon stimulation) did not change significantly. With regard to the HDL2-cholesterol and 18:0 fatty acid concentration in serum the group can be divided into responders (with a decline of glycosylated haemoglobin) and non-responders. The two groups have a reverse trend of blood sugar levels and insulinaemia and differ as to the metabolism of 18:1 n-7 acid. CONCLUSIONS: Enrichment of the diet with n-3 fatty acids in diabetics with dyslipidaemia has a favourable effect on the plasma lipid spectrum without causing deterioration of parameters of diabetes compensation. Among the group of patients some can be found where fish oil administration improves also glucose homeostasis.

[Determination of thyroid microsomal antibodies in the blood of patients with thyropathies using the ELISA method]. / Stanovení protilátek proti mikrosomum stítné zlázy v sérech pacientu s tyreopatiemi metodou ELISA.

The authors elaborated and tested an ELISA method for the assessment of antibodies against the microsomal fraction of the human thyroid gland (Thyroid microsomal antibodies). The method is more sensitive than passive haemagglutination. The reproducibility is satisfactory, the intraassay variation coefficient is 10.7%, the interassay variation coefficient 15.4%. In a group of 282 subjects with thyropathies a positive TMAb was found in 57.8%, in a control group of 63 subjects in three instances (4.7%). The method makes it possible to assess individual classes of specific immunoglobulins--IgG in the group of 15 positive sera accounted for 78% of all immunoglobulins. The importance of IgM and IgA is less clear. The method is suited in particular for screening of autoimmune affections of the thyroid gland.

[A minimal model of insulin and glucose kinetics]. / Minimální model kinetiky inzulínu a glukózy.

The minimal model of insulin and blood sugar kinetics is a method the basis of which are two mathematical models describing the dynamic changes of plasma glucose and insulin concentration along with the beta-cell response to the rise of the blood sugar level during the intravenous glucose tolerance test with frequent collection of samples. By means of computer processing the following indices are evaluated: SI--insulin sensitivity, SG--efficacy of glucose, phi 1 and phi 2 describing the sensitivity of beta-cells for glucose. According to present knowledge it is a simple method with a satisfactory reproducibility and validity of assembled results, which is safe for the examined subject and unpretentious as regards apparatus. It makes it possible to evaluate the glucose and insulin relationship in their mutual dynamic relationship and to create specific modifications as well as the follow-up of other indicators.