7-oxygenated derivatives of dehydroepiandrosterone and obesity.

7-hydroxy/oxo derivatives of dehydroepiandrosterone are potential regulators of the local cortisol activity due to their competition in the cortisolcortisone balance mediated by 11ß-hydroxysteroid dehydrogenase. 7-hydroxy-dehydroepiandrosterone is marketed as anti-obesity medication, though no clinical study aimed at the benefit of administering 7-oxygenated derivatives of dehydroepiandrosterone has appeared until now. We tried to show whether there exist differences in levels of circulating 7-hydroxy/oxo-dehydroepiandrosterone derivatives between lean and obese boys and girls. From a cohort of adolescents investigated within the frame of anti-obesity programme 10 obese boys and 10 obese girls were compared with age-matched lean boys and girls in their anthropometric data, and concentrations of both epimers of 7-hydroxy-dehydroepiandrosterone and 7-oxo-dehydroepiandrosterone were determined by the RIA method. The basal levels of 7α-hydroxy-dehydroepiandrosterone were significantly higher in obese boys than in lean boys but not in girls. The association was found for anthropometric parameters and 7α-hydroxy-dehydroepiandrosterone, however again only in boys and not in girls. Higher levels of 7α-hydroxy-dehydroepiandrosterone its positive association with anthropometric data in obese boys may serve as a sign that, at least in boys, 7-oxygenated 5-ene-steroids may take part in regulating the hormonal signal for fat formation or distribution.

Anti-Helicobacter Pylori, anti-thyroid peroxidase, anti-thyroglobulin and anti-gastric parietal cells antibodies in Czech population.

Autoimmune thyropathies are frequently linked to many infections, such as Helicobacter pylori, which are also supposed to play a role in their pathogenesis. The aim of this study was to evaluate the relationships between thyroid and gastric autoimmunity and H. pylori infection on a large sample of Czech population (n=1621) by monitoring the autoantibodies against thyroglobulin (anti-Tg) and thyroid peroxidase (anti-TPO) and gastric parietal cell (anti-GPC, representing thyrogastric syndrome) in correlation with antibodies against Helicobacter pylori (anti-H. pylori) of classes IgG and IgA. The interrelation between autoantibodies and H. pylori antibodies was assessed by H. pylori seropositivity. In H. pylori seropositive persons as compared to seronegative irrespective of age and sex, a higher occurrence of anti-TPO (10.4 % vs. 5.8 %, p=0.001) and anti-GPC (6.1 % vs. 1.7 %, p<0.001) was found. Differences in anti-TPO occurrence were significant in both men (7.0 % vs. 3.3 %, p=0.03) and women (12.7 % vs. 8.0 %, p=0.02), differences in anti-GPC occurrence were significant only in women (7.2 % vs. 1.7 %, p<0.001). Results of this study support the idea of a connection between infection of H. pylori and the occurrence of anti-TPO autoantibodies representing thyroid autoimmunity and gastric parietal cells autoantibodies representing the thyrogastric syndrome.

Immunoprotective steroids and SHBG in non-treated hypothyroidism and their relationship to autoimmune thyroid disorders.

Immunomodulatory steroids, dehydroepiandrosterone and its 7-hydroxylated metabolites and sex hormone-binding globulin (SHBG) were determined in sera of 88 women aged 18-75 years. The group consisted of 34 healthy women, 37 women with subclinical and 17 women with manifest hypothyroidism. In all subjects the laboratory parameters of thyroid function (thyrotropin, free thyroxine and triiodothyronine) and thyroid autoantibodies to thyroid peroxidase and thyroglobulin were determined. The aim was to find out 1) whether the above steroids and SHBG levels differ in individual groups according to thyroid status, 2) whether correlations exist among investigated steroids and thyroid laboratory parameters, and 3) whether the respective steroid and SHBG levels differ according to the presence of principal thyroid autoantibodies. With the exception of 7beta-hydroxy-dehydroepindrosterone levels, which were decreased in patients with manifest hypothyroidism (p<0.05), no significant differences in steroid and SHBG levels among groups according to diagnosis were found. On the other hand, significantly decreased levels of all the immunomodulatory steroids studied were found in subjects with positive titres of thyroid autoantibodies. This finding was supported by a tight negative correlation among the above steroids and thyroid autoantibodies. In addition, these steroids correlated negatively with thyrotropin and positively with free thyroid hormones. The results point to a negative relationship between the above mentioned immunoprotective steroids and the extent of the autoimmune process in hypothyroidism.

Fasting insulin pulsatile secretion in lean women with polycystic ovary syndrome.

The aim of our study was to evaluate rapid insulin pulses and insulin secretion regularity in fasting state in lean women with polycystic ovary syndrome (PCOS) in comparison to lean healthy women. PCOS (n=8) and controls (n=7) underwent every minute blood sampling for 60 min. Insulin pulsatility was assessed by deconvolution and insulin secretion regularity by approximate entropy methodology. PCOS had higher testosterone (p<0.02), prolactin (p<0.05) and lower sex hormone binding globulin (SHBG) (p<0.0006) levels than controls. Approximate entropy, insulin pulse frequency, mass, amplitude and interpulse interval did not differ between PCOS and controls. PCOS had broader insulin peaks determined by a common half-duration (p<0.07). Burst mass correlated positively with testosterone (p<0.05) and negatively with SHBG (p 0.0004) and common half-duration correlated positively with prolactin (p<0.008) and cortisol levels (p<0.03). Approximate entropy positively correlated with BMI (p<0.04) and prolactin (p<0.03). Lean PCOS patients tended to have broader insulin peaks in comparison to healthy controls. Prolactin, androgens and cortisol might participate in alteration of insulin secretion in PCOS-affected women. Body weight and prolactin levels could influence insulin secretion regularity.

The influence of galvanic currents and voltage on the proliferation activity of lymphocytes and expression of cell surface molecules.

Release of metal ions from dental metal fillings supported by galvanism can cause local or general pathological problems in sensitive and genetically susceptible individuals. We aimed to investigate in vitro lymphocyte responses and expression of surface molecules influenced by galvanic currents and voltage. Human peripheral blood lymphocytes were influenced by galvanic currents and voltages and lymphocyte proliferation was measured. Control samples were not exposed to the influence of galvanism. We also studied the expression of surface molecules by the FACS analysis. A 15-h and shorter influence of almost all tested currents and voltages caused a significant decrease in lymphocyte proliferation and the 15-h influence of 20 microA currents significantly increased expression of surface molecules CD 19, 11a/18, 19/69 and 19/95. An influence of 10 and 3 microA currents led to a significant decrease in the expression of surface molecules CD 3, 11a/18, 3/69 and 3/95 and to a significant increase in CD 19 expression. An 80 mV voltage influence led to a significant decrease in the expression of surface molecules CD 3, 11a/18, 3/69, 3/95, 19/69 and 19/95, and 200 and 300 mV voltages significantly decreased the expression of surface molecules CD 3, 19, 11a/18, 3/95 and 19/95 and significantly increased CD 19/69 expression. A long-lasting influence of galvanism can, in sensitive and genetically susceptible individuals, influence lymphocyte proliferation and surface molecule expression. The threshold for pathological values of 5 microA for galvanic currents and 100 mV for galvanic voltage was confirmed.

Helicobacter pylori isolated from patients with tonsillar cancer or tonsillitis chronica could be of different genotype compared to isolates from gastrointestinal tract.

Helicobacter pylori from patients with different diseases, including so-called autoimmune thyroiditis, chronic tonsillitis and tonsillar cancer, was isolated and cultured. It was identified according to the genotype using labeled hybridization probes complementary to six sequences of cagA and vacA genes. Different types of strains were found in isolates from gastrointestinal tract and patients suffering from thyroiditis. Six out of seven genotyped isolates from patients in our Department of Otorhinolaryngology and Head and Neck Surgery exhibited the same genotype, differing from isolates obtained from other patients; the 7th isolate originated from a patient who had undergone surgery for deviatio septi nasi, at the same time suffering from autoimmune thyroiditis, having confirmed gastric infection by H. pylori from biopsy. This data made it possible to formulate the hypothesis on probable association of specific H. pylori genotype with chronic tonsillitis and tonsillar cancer. We assessed commercial transport media and improved nucleic acid isolation techniques and the RT-PCR-based tests, which allowed us to skip a culture step and to test directly the patients' samples; however, for full confirmation of our hypothesis and explanation of possible mechanisms of the contribution of Helicobacter sp. to the pathogenesis of the disease further data are to be collected and evaluated.

[Polyglandular activation of autoimmnunity as a manifestation of subclinical endocrinopathies]. / Polyglandulární aktivace autoimunity jako projev subklinických endokrinopatií.

BACKGROUND: Autoimmune thyropathies belong to the most frequently occurring autoimmune endocrinopathies. Autoimmune thyropathies occur either independently or linked to known polyglandular syndromes of type I-III. METHODS AND RESULTS: During the last decade, we observed a group of patients with autoimmune thyroiditis, in which autoimmune endocrinopathies were mutually associated and named the symptoms of this group "polyglandular activation of autoimmunity". The frequency of the occurrence of autoantibodies against other endocrine organs in this group was determined and the most frequent was the occurrence of the autoantibodies against steroid producing cells - anti-ovary 28 %, anti-adrenal 23 %, anti-testes 12 %. Considering the most frequent occurrence of autoantibodies against steroid producing cells, attention has been paid namely to patients with autoimmune thyroiditis and a concurrent occurrence of anti-adrenal autoantibodies. In the foreground of the clinical picture of these patients were dysregulations on the metabolic and circulation levels together with symptoms of discomfort (subfebrile condition, arthralgia and fatigue). Heavy fatigue of these patients was linked to the changes of levels and mutual ratio of melatonin and serotonin and regarding autoantibodies, mainly autoantibodies against steroid producing cells, namely against the particular cells of the layers of adrenal cortex played a role. CONCLUSIONS: The presence of autoantibodies influenced also the functional response, namely the ACTH/cortisol ratio. Autoantibodies detected were not anti-21-hydroxylase autoantibodies, typical for autoimmune polyglandular syndrome II, but antibodies against antigens of other molecular weight.

[Endocrine orbitopathy and significance of autoantibodies against 1D protein]. / Endokrinní orbitopatie a význam autoprotilátek proti 1D proteinu.

BACKGROUND: Endocrine ophthalmopathy is a chronic eye disease, characterized by inflammation in parabulbar and retrobulbar space, occurring usually in Graves' thyrotoxicosis. Although the pathogenesis of the disease has not been clarified until now, it is accepted that this disease is of an autoimmune nature, where the targets of the autoimmune reaction are the antigens shared by thyroid and orbit-tissue. The autoantibodies against recombinant 1D protein are highly specific and sensitive for the diagnosis of endocrine orbitopathy. METHODS AND RESULTS: The aim of our study was to establish, whether the autoantibodies against 1D protein are found predominantly in patients with clinically expressed endocrine orbitopathy. We evaluated in 30 patients with clinically expressed endocrine orbitopathy the thickness of the three retrobulbar eye muscles, damaged by endocrine orbitopathy, determined the parameters of thyroid hormones and anti-TSH receptor autoantibodies. In all patients the detection of circulating autoantibodies against recombinant 1D protein was performed. Autoantibodies against recombinant 1D protein were found in all patients with clinically expressed endocrine orbitopathy. CONCLUSIONS: Immunoreactivity did not depend on the duration or severity of the eye disease, neither on patients' age. We did not find any correlation between the thickness of eye muscles and the titre of anti-TSH receptor autoantibodies, levels of ssTSH and free thyroxine and also any correlation between the thickness of eye muscles and the disease duration.

Vitamin D supplementation changed relationships, not levels of metabolic-hormonal parameters in autoimmune thyroiditis.

In women with chronic autoimmune thyroiditis and vitamin D deficiency we have found reference levels of relevant metabolic-hormonal parameters except for parathormone and total calcium. Three months supplementation with vitamin D (4300 IU/day, cholekalciferol) did not lead to significant changes of investigated hormonal parameters, while the levels of parathormone and calcium reached normal levels. However, a correlation analysis revealed marked changes in mutual relations. First, an inverse correlation of vitamin D with parathormone, insulin secretion (C peptide, insulin) and its efficiency (HOMA IR) disappeared. Relationships of vitamin D to hepatic insulin resistance (insulin/C peptide), to DHEA (both negative), and to DHEAS/DHEA ratio (positive) were newly found. Second, a positive correlation of CRP with insulin secretion remained, while its relation to insulin efficiency (HOMA IR, insulin/C peptide) was newly observed. Analogical positive correlations appeared also among anti TPO and insulinemia, insulin/C peptide, HOMA IR, and anti Tg to C peptide. A relationship of the CRP with anti TPO became significant (+). Third, out of glucose metabolism parameters only insulin/C peptide and glycemia did not correlate with vitamin D during its deficiency, while after supplementation insulin/C peptide alone correlated positively with both DHEAS and DHEA, and negatively with vitamin D.

[Autoimmune thyroiditis--selected etiopathogenic mechanisms]. / Autoimunitní tyreoiditida--vybrané etiopatogenetické mechanizmy.

Autoimmune thyroiditis occurs as organ specific autoimmune disease not only as an isolated impairment of thyroid gland, but also linked to many autoimmune endocrinopathies. Genetic predisposition in the area of HLA antigens was followed up by patients with autoimmune thyroiditis diagnosed in this way and it appeared that genetic predisposition in isolated autoimmune thyroiditis is different when compared to the occurrence linked to endocrine polyglandular disease. In selected groups of patients with autoimmune disease also the influence of extraneous factors on the development of the autoimmune process was followed up, namely the influence of heavy metals and the influence of infectious agent--Helicobacter pylori. These factors have a different character of activation of autoimmune thyroiditis too, depending on the character of its manifestation as isolated disorder or in link to autoimmune polyglandular syndrome type II, or in link to the group of polyglandular activation of autoimmunity. To conclude, this study leads to the assumption, that autoimmune thyroiditis is a set of clinical syndromes that depends on the activation of the autoimmune process, rather than a strictly genetically and epigenetically characterized nosological unit.

Levels of adipokines and some steroids during the menstrual cycle.

The cyclical effects of hormones during the menstrual cycle (MC) are not just responsible for driving ovulation, but also have significant influence on dietary intake and appetite, as well as psychological and behavioral changes. The aim of our study was to describe changes and relationships between the MC and selected steroids, adipokines and food intake-related hormones. Twenty-seven women with regular menstrual cycles were included in the study, and their hormonal spectrum was measured in regular intervals starting from the first day of their cycle. Classical changes in gonadotropins, estrogens and progesterone during the menstrual cycle are accompanied by less striking but significant changes in 17-hydroxyprogesterone and testosterone. No significant changes show dehydroepiandrosterone and its 7-oxygenated metabolites. Adipokines show a tendency to increase during ovulation, while ghrelin and resistin decrease. There is also a remarkable association of sex hormone-binding globulin on the day of the cycle. Our results demonstrate that changes to adipokines during the menstrual cycle are not substantial, but nonetheless can play a role in the changes of food intake described in the literature. Precise descriptions of physiological changes in healthy women are important in helping us understand the significance of the changes accompanying various pathological states.

Comparison of cytokine levels in sera of patients with autoimmune endocrinopathies.

Autoimmune endocrinopathies can be divided according to the presence of organ specific autoantibodies and according to the clinical manifestations into isolated autoimmune endocrinopathies, autoimmune polyglandular syndromes (APS) and polyglandular activation of autoimmunity (PAA). Many factors take part in the development of the autoimmune disease: predominantly a genetic predisposition, environmental etiologic causes and dysregulation in the microenvironment of the target organ. Until now it is not completely clear, if manifestations of the clinical disease depend primarily upon external factors and the degree of regulation mechanism disorder (e.g. in Th1/Th2 regulation) or upon the different genetic predisposition. In this work we compared the levels of Th1 and Th2 lymphocyte cytokines in peripheral blood in three groups of patients: group A of 30 patients with autoimmune thyroiditis, group B of 25 patients with PAA, and group C of 10 patients with APS type II. From group of Th1 cytokines IL-2 and IFN-gamma were detected, whereas from group of Th2 cytokines IL-4 and IL-10 were determined by ELISA kit. We did not find any differences in the concentrations of IL-2, IFN-gamma, IL-4 and IL-10 among the groups of patients with autoimmune endocrinopathies.

HLA antigen expression in autoimmune endocrinopathies.

The HLA allelic frequency was determined in three groups of autoimmune endocrinopathies: A) 30 patients with autoimmune thyroiditis, B) 20 patients with polyglandular activation of autoimmunity, and C) 10 patients with the autoimmune polyglandular syndrome type II. The groups were defined by the clinical state and serological parameters. Healthy blood donors of Caucasian population from the US database of HLA frequencies served as the controls. In group A, a higher occurrence of HLA-A24 (21.7 %) was found as compared to group B (5.0 %) and to the controls (8.5 %), of HLA-B27 (15.0 %) and of HLA-DR-11 (20 %) as compared to the controls (4.2 % and 8.5 %). In group B, a higher occurrence of HLA-A3 (25.0 %) was found as compared to group A (10 %) and to the controls (11.8 %), and of HLA-B8 (22.5 %) as compared to group A (8.3 %) and to the controls (8.6 %). In this group the occurrence of HLA-DR3 (30.0 %) was higher as compared to group A (10.0 %) and to the controls (9.8 %) and of HLA-B8 (30.0 %) as compared to group A (8.3 %) and to the controls (8.6 %). Genetic markers indicate a similarity of groups B and C. Patients in these groups could be at different stages of the same disease, however, some distinctions between them lead us to consider the possibility whether different epigenetic factors could extend the difference between these groups in the course of clinical development.

Serum levels of IGF-I, HGF, TGFbeta1, bFGF and VEGF in thyroid gland tumors.

IGF-I, HGF, TGFbeta1, bFGF and VEGF are involved in the pathogenesis of thyroid gland tumors and their growth. We decided to find whether changes in the production of these cytokines by thyroid tumor cells are reflected by changes of their peripheral blood. Using ELISA kits, we measured the concentrations of growth factors in the peripheral blood serum in 28 patients with thyroid gland tumors (14 adenomas, 14 papillary carcinomas) and compared these concentrations with those in healthy people. We found significantly lower serum levels of IGF-I in patients with thyroid adenoma compared to the healthy population. Serum levels of HGF and bFGF were significantly higher in patients with thyroid adenoma and papillary carcinoma compared with those in healthy subjects. Serum concentrations of TGFbeta1 and VEGF were not significantly different in any groups of investigated subjects. Changes in the production of these cytokines by thyroid gland tumor cells are reflected in their peripheral blood levels, but these levels also depend on a number of other physiological and pathological processes in the organism. However, significant differences of HGF and bFGF serum levels can be explained by their very high production by thyroid tumor cells and by their strong effect on the follicular and endothelial cell proliferation.

[Extra-thyroid autoantibodies in autoimmune thyroiditis]. / Extratyroideální autoprotilátky u autoimunitní tyroiditidy.

Organ specific autoimmune diseases do not occur isolated. In the authors group of 4509 patients with confirmed autoimmune thyroiditis the authors investigated the frequency of extrathyroid organ specific and non-specific autoantibodies. The objective was to draw attention to other autoimmune diseases which may not yet be clinically manifest. The mutual positivity of antibodies against thyroid peroxidase (TPO) and antibodies against the adrenals was: in the zona glomerulosa 21.1%, in the zona fasciculata 3.0%, in the zona reticularis 19.5%, in the adrenal medulla 9.0%. The mutual positivity of antibodies against TPO and ovaries was 28.5%, testes 12.5%, parathyroid 10.2%, islets of Langerhans 8.2%, hair follicles 45.0%, cell nuclei 26.1%, mitochondria 3.1%, smooth muscles 2.4%, striated muscle 0.2%, and gastroparietal cells of the stomach 12.3%. These results draw attention to the importance of polyclonal activation of the autoimmune process in endocrinopathies and the necessity to assess extrathyroid autoantibodies in autoimmune thyroiditis.

[The fatigue syndrome in autoimmune thyroiditis with polyglandular activation of autoimmunity]. / Unavový syndrom u autoimunitní tyroiditity s polyglandulární aktivací autoimunity.

The authors compared in a group of 118 patients with autoimmune thyroiditis and a positive antibody titre against ovaries the grade of fatigue with the presence of organ specific and non-specific autoantibodies in the peripheral blood stream, antibodies against EBV and CMV, immunoglobulin concentrations, biochemical parameters of the lipid metabolism, glucose tolerance, ion balance and melatonin and serotonin levels. Patients with autoimmune thyroiditis were differentiated according to the degree of fatigue into three groups: 38 with fatigue typical for CFS, 30 with occasional fatigue and 50 without the feeling of fatigue. Fatigue of the CSF type was characterized by a significantly higher incidence of autoantibodies against the adrenals and a higher cholesterol level. Increased fatigue of the patients was associated with a lower melatonin level, a higher serotonin level and a lower M/S ratio as compared with patients without fatigue. In other indicators no differences were found. Fatigue in CFS could be associated, similarly as in autoimmune endocrinopathies, with impaired immunoendocrine regulation. In autoimmune thyroiditis, regardless of the concomitant presence of fatigue, in addition to antibodies against thyroid peroxidase most frequently antibodies against the ovaries were detected.

[Reactions to metals in patients with chronic fatigue and autoimmune endocrinopathy]. / Reakce na kovy u pacientu s chronickou únavou a autoimunitními endokrinopatiemi.

Our study was designed to assess the effect of heavy metals on the severity of fatigue in autoimmune thyroid disease associated with autoantibodies against other endocrine organs. We compared our data with those obtained from other groups of patients. A total of five groups of patients were examined by their medical history, dental examination, and using a modified test of blast transformation of metals (Melisa): a) 10 fatigues female patients with autoimmune thyroidism and polyglandular activation of autoimmunity, b) 12 fatigued patients with autoimmune thyroidism, c) 28 fatigued patients free of endocrinopathy, d) 22 professionals without evidence of autoimmunity, e) 13 controls, a population sample, the individuals did not complain of marked fatigue and their laboratory tests did not show signs of autoimmunity and endocrinopathy. Fatigue regardless of the underlying disease is primarily associated with hypersensitivity to inorganic and organic mercury, nickel, and gold. The groups differed in their hypersensitivity to other metals. In the control group, hypersensitivity--mostly to cadmium and lead--was found in four of the examined individuals only. Statistical analysis of data obtained from professionals and controls revealed a higher incidence of positivity to organic and inorganic mercury and nickel in professionals.

New insights into the physiology of bone regulation: the role of neurohormones.

Bone metabolism is regulated by interaction between two skeletal cells - osteoclasts and osteoblasts. Function of these cells is controlled by a number of humoral factors, including neurohormones, which ensure equilibrium between bone resorption and bone formation. Influence of neurohormones on bone metabolism is often bimodal and depends on the tissue, in which the hormone is expressed. While hypothalamic beta-1 and beta-2-adrenergic systems stimulate bone formation, beta-2 receptors in bone tissue activate osteoclatogenesis and increases bone resorption. Chronic stimulation of peripheral beta-2 receptors is known to quicken bone loss and alter the mechanical quality of the skeleton. This is supported by the observation of a low incidence of hip fractures in patients treated with betablockers. A bimodal osteo-tropic effect has also been observed with serotonin. While serotonin synthetized in brain has osteo-anabolic effects, serotonin released from the duodenum inhibits osteoblast activity and decreases bone formation. On the other hand, both cannabinoid systems (CB1 receptors in the brain and CB2 in bone tissue) are unambiguously osteo-protective, especially with regard to the aging skeleton. Positive (protective) effects on bone have also been shown by some hypophyseal hormones, such as thyrotropin (which inhibits bone resorption) and adrenocorticotropic hormone and oxytocin, both of which stimulate bone formation. Low oxytocin levels have been shown to potentiate bone loss induced by hypoestrinism in postmenopausal women, as well as in girls with mental anorexia. In addition to reviewing neurohormones with anabolic effects, this article also reviews neurohormones with unambiguously catabolic effects on the skeleton, such as neuropeptide Y and neuromedin U. An important aim of research in this field is the synthesis of new molecules that can stimulate osteo-anabolic or inhibiting osteo-catabolic processes.

Simple approach for analysis of plasma oxo-, hydroxy- and dicarboxylic acids.

Following deproteinization of plasma with organic solvents the supernatant was shaken with hexane and cation-exchange resin in an Eppendorf tube to remove fatty and amino acids and the medium was subjected to direct treatment with ethyl chloroformate under catalytic influence of pyridine. A subsequent extraction of the immediately formed ethyl esters with a drop of chloroform enabled us to subject the sample to gas chromatographic (GC) analysis. Since ketocarboxylic acids do not require a preliminary oximation the total time of sample workup and analysis takes only several minutes.