Maria Klatka

Th17 and Treg cells in adolescents with Graves' disease. Impact of treatment with methimazole on these cell subsets.

Abstract The role of T helper 17 (Th17) and T regulatory cells (Treg) in the pathogenesis of Graves’ disease (GD) remains uncertain. The influence of methimazole (MMI) on the human immune system is still poorly understood. The aim of the present research was to assess changes in the frequencies of peripheral blood Th17 and Treg cells during GD treatment in the group of teenagers. The frequencies of Th17 and Treg were measured by flow cytometry in 60 adolescents at the time of GD diagnosis and after achieving MMI-induced euthyreosis. The control group consisted of 20 healthy volunteers. Lower percentages and absolute counts of Treg cells were found in the study group before the treatment in comparison with healthy controls (p = 0.032 and p = 0.006, respectively). Treatment with MMI caused an increase in the percentages and absolute counts of Treg lymphocytes (p = 0.037 and p = 0.007). After the treatment, no clinically significant differences in Treg cells between GD patients and controls were found. Higher absolute counts of Th17 lymphocytes were found in hyperthyroid adolescents before the treatment initiation and after achieving euthyreosis than in healthy individuals (p = 0.0001 and p = 0.047). Treatment with MMI caused a significant decrease in the percentages and absolute counts of Th17 lymphocytes (p = 0.047 and p = 0.043). The present study demonstrates that both Th17 and Treg cells might play a role in the pathogenesis of GD. Increased percentage of Treg after MMI therapy seems a predictor of response to anti-hypertensive treatment as it is associated with the normalization of thyroid hormone levels.

Serum and urinary selenium levels in obese children: a cross-sectional study

OBJECTIVE To determine serum and urinary selenium (Se) levels in children with and without obesity, and to assess if Se influences the risk of obesity. SUBJECTS AND METHODS High-resolution-continuum source-atomic absorption spectrometry (HR-CS-AAS) was used to determine the content of Se in 80 children (age 6-17; 40 boys, 40 girls). Correlations between variables were tested with the use of Spearmans correlation coefficient. U Mann-Whitney test was applied to assess the difference of Se contents in samples. Measured metabolic risk factors (blood pressure, glucose level, triglycerides (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), and total cholesterol), age, gender, and BMI were correlated. Logistic regression models were fitted to identify predictors of obesity interacting with selenium content in serum and urine, separately. RESULTS Obese children, regardless of gender, had lower Se content. Se level in serum (p=0.001, OR 0.74, 95%CI 0.62-0.88) and total cholesterol (p=0.001, OR 1.19, 95%CI 1.08-1.31) were the independent factors significantly influencing the risk of obesity in children. Two separate models were observed for Se in urine: (i) Se level (p<0. 0001, OR 0.70, 95%CI 0.58-0.84) and glucose level (p<0.0001, OR 1.22, 95%CI 1.10-1.35), and (ii) Se level (p=0.002, OR 0.60 95%CI 0.43-0.83) and total cholesterol level (p=0.003, OR 1.16, 95%CI 1.05-1.28). CONCLUSION The current study suggests a possible role of Se in obesity. Further research needs to be performed to check if obese children are an at-risk group for Se deficiency.

Determination of total iodine in serum and urine samples by ion chromatography with pulsed amperometric detection - studies on analyte loss, optimization of sample preparation procedures, and validation of analytical method.

A fast, accurate and precise ion chromatography method with pulsed amperometric detection was applied to evaluate a variety of parameters affecting the determination of total iodine in serum and urine of 81 subjects, including 56 obese and 25 healthy Polish children. The sample pretreatment methods were carried out in a closed system and with the assistance of microwaves. Both alkaline and acidic digestion procedures were developed and optimized to find the simplest combination of reagents and the appropriate parameters for digestion that would allow for the fastest, least time consuming and most cost-effective way of analysis. A good correlation between the certified and the measured concentrations was achieved. The best recoveries (96.8% for urine and 98.8% for serum samples) were achieved using 1ml of 25% tetramethylammonium hydroxide solution within 6min for 0.1ml of serum/urine samples. Using 0.5ml of 65% nitric acid solution the best recovery (95.3%) was obtained when 7min of effective digestion time was used. Freeze-thaw stability and long-term stability were checked. After 24 weeks 14.7% loss of iodine in urine, and 10.9% in serum samples occurred. For urine samples, better correlation (R(2)=0.9891) of various sample preparation procedures (alkaline digestion and application of OnGuard RP cartidges) was obtained. Significantly lower iodide content was found in samples taken from obese children. Serum iodine content in obese children was markedly variable in comparison with the healthy group, whereas the difference was less evident when urine samples were analyzed. The mean content in serum was 59.12±8.86μg/L, and in urine 98.26±25.93 for obese children when samples were prepared by the use of optimized alkaline digestion reinforced by microwaves. In healthy children the mean content in serum was 82.58±6.01μg/L, and in urine 145.76±31.44μg/L.

Peripheral blood lymphocyte apoptosis and its relationship with thyroid function tests in adolescents with hyperthyroidism due to Graves' disease.

Introduction Failures in apoptotic pathways can contribute to various autoimmune diseases, including autoimmune hyperthyroidism due to Graves’ disease (GD). The aim of the present research was to assess changes in the degree of peripheral blood (PB) lymphocyte apoptosis during methimazole (MMI) treatment in the group of teenage children, and to describe its relationship with thyroid function tests. Material and methods The percentage of PB apoptotic lymphocytes, assessed by the decrease in mitochondrial transmembrane potential (CMXRos staining), was measured in 30 adolescents at the time of diagnosis and after obtaining normalization of the thyroid hormone levels. Results The percentage of apoptotic lymphocytes in previously untreated patients with GD (5.16 ±2.81%) was significantly lower (p = 0.000001) than the percentage of apoptotic cells in the same group of patients after obtaining methimazole-induced euthyroidism (10.72 ±4.66%). There was a correlation between the increase of the mean percentages of apoptotic lymphocytes and the reduction of FT4 levels (R = 0.63, p < 0.0001), as well as the reduction of TT3 levels (R = 0.95, p < 0.0001). The more signs and symptoms accompanying the diagnosis of GD, the higher was the increment of the degree of lymphocyte apoptosis observed during the MMI-treatment (R = 0.74, p < 0.0000001). The methimazole dosage correlated (R = 0.85, p < 0.0001) with the percentage of apoptotic cells. Conclusions The use of methimazole in treatment of hyperthyroidism due to GD leads to an increment of apoptotic cells in PB. Higher doses of methimazole cause a higher increase of apoptotic lymphocytes. Apoptosis induction of human PB lymphocytes seems to be one of the indicators of proper hyperthyroidism treatment.

The Influence of Intracerebral Streptozotocin and/or Cadmium on Memory Processes in Mice Exposed to Transient Cerebral Oligemia

The aim of the study was to examine the effects of simultaneous acute exposure to cadmium (Cd) and in the presence of streptozotozin (STZ) on the central nervous system (CNS) memory processes in mice subjected to transient brain ischemia. In order to obtain transient brain ischemia, operative occlusion of common carotid arteries for 30 min was performed. Cadmium chloride was administered intraperitoneally (ip) after the surgery at a single dose of 0.1 LD50 (LD50 = 14 mg/kg). Disturbances in glucose metabolism in the brain tissue were induced by bilateral intracerebral administration of STZ, a drug that inhibits the function of a neuronal insulin receptor. Long-term memory was evaluated by means of a step-through passive avoidance task. Spatial working memory expressed as spontaneous alternations was tested in the Y-maze test. Coexposure to brain oligemia and STZ on the CNS produced significant impairment of long-term memory processes in mice. An additional exposure to Cd exacerbated the deficits of these processes. These results indicate that brain oligemia, Cd, and altered glucose metabolism may aggravate adverse effects on memory.

SHORT syndrome in a two-year-old girl – case report

BackgroundSHORT syndrome is a rare genetic congenital defects condition. The frequency of the disease still remains unknown.Case presentationWe report the two-year-four-month old female with SHORT syndrome who present growth retardation and dysmorphic features (triangular-shaped face, prominent forehead, ocular depression, lipodystrophy at the lumbar region and around elbows), consistent with the phenotype described for this syndrome.The molecular analysis showed the presence of heterozygous variant c.1956dupT (p.Lys653*) in exon 15 of PIK3R1.ConclusionsThe frequency of the disease still remains unknown; solely several dozen cases have been described worldwide.

Expression of CD200 and CD200R regulatory molecules on the CD83+ monocyte-derived dendritic cells generated from patients with laryngeal cancer.

CD200 molecule may play a role in local tumor invasion and augmenting the metastatic capacity of squamous cell carcinoma. The objective of the study was to assess by means of flow cytometry the expression of CD200 and its receptor, CD200R, on CD83+ monocyte-derived dendritic cells (Mo-DCs), pulsed or not with autologous tumor cell lysates (aTCL) in patients who suffer from laryngeal carcinoma in comparison to healthy donors. The median value of CD200 mean fluorescence intensity (MFI) on the Mo-DCs pulsed with aTCL of the patients with laryngeal cancer was 61.94 and was statistically significantly higher than on the unpulsed Mo-DCs of these patients (24.81) and healthy donors (16.63), p = 0.0034 and p = 0.0004, respectively. Median MFI score of CD200R in specimen derived from patients with laryngeal cancer was 259.31 on Mo-DCs pulsed with aTCL, while in unpulsed Mo-DCs was 86.74 (p = 0.0035) and on the Mo-DCs from control group it was 67.51 (p = 0.0004). The obtained results showed a relation between the presence of laryngeal cancer and the expression of CD200 and CD200R molecules on the CD83+ Mo-DCs pulsed with autologous cancer cell lysates. This analysis may have implications for setting new therapeutic options for cancer immunotherapy in the future.

Impact of Methimazole Treatment on Magnesium Concentration and Lymphocytes Activation in Adolescents with Graves’ Disease

The aim of this research was to assess plasma magnesium (Mg) concentration, the frequencies of activated T CD4+ and T CD8+ lymphocytes and B lymphocytes in adolescents with hyperthyroidism due to Graves’ disease (GD), and to assess changes in the above-mentioned parameters during methimazole (MMI) treatment. The frequencies of activated T and B cells were measured by flow cytometry method and plasma Mg concentration was determined by spectrophotometry method in 60 adolescents at the time of GD diagnosis and after receiving the normalisation of the thyroid hormones levels. The control group consisted of 20 healthy volunteers. We observed lower plasma Mg concentration, and higher frequencies of activated T and B lymphocytes in the study group before the treatment in comparison with healthy controls, and with study group in MMI-induced euthyreosis (p < 0.01).Statistically significant negative correlations between the percentages of activated T CD3+, T CD4+, T CD8+ and B CD19+ lymphocytes, and plasma Mg concentration before the treatment were found (r < −0.335, p < 0.002). After the treatment no vital differences in plasma Mg concentration, and in percentages of activated cells between GD patients and controls were found, except CD8+CD25+ cells (p = 0.03). The present study demonstrates that both activated T and B cells might play an important role in the pathogenesis of GD, and activation is related to Mg plasma level. The use of MMI in treatment of hyperthyroidism due to GD leads to decrease the frequencies of activated lymphocytes and normalisation of Mg levels.

Influence of growth hormone therapy on selected dental and skeletal system parameters.

INTRODUCTION Growth hormone deficiency (GHD) is one of the main indications for growth hormone therapy. One characteristic of this disease is bone age delay in relation to the chronological age. Pituitary dysfunction negatively affects the growth and development of the jaws and teeth of the child. The secretion of endocrine glands regulates growth, development, and gender differentiation. It also controls the growth of bones and teeth, regulates metabolism of calcium and phosphate, proteins, lipids and carbohydrates. The primary role in the endocrine system is played by the pituitary gland which is responsible for the production of somatotropin [1]. Dysfunction of the pituitary gland has a negative effect on the growth and development of long bones in the body, and may have an adverse effect on the development of maxilla, mandible and dentition of a child. There is some information in the literature that dental age is delayed in short stature children; the replacement of deciduous teeth by permanent teeth is also delayed, and newly erupted permanent teeth often require orthodontic treatment. Applying hormonal therapy positively affects the process of replacement of dentition [2, 3, 4, 5, 6]. OBJECTIVES The aim of the study was to assess bone and dental age, as well as analyze the state of dentition in children diagnosed with GH deficiency treated with growth hormone, depending on the duration of treatment. MATERIAL AND METHODS The study material consisted of 110 children (27 males, 83 females), hospitalized for somatotropin hypopituitarism in the Department of Paediatric Endocrinology and Diabetology at the Medical University of Lublin, Poland. The mean birth age was 13 years (156 months) with a standard deviation of 2 years and 6 months (30 months). 47 children (43%) started treatment with the growth hormone (group starting treatment) and 63 children (57%) whose treatment was started 2-3 years previously (group in the course of treatment). The control group consisted of 41 generally healthy children (15males, 25 females) with ENT problems, such as hypoacusis and a condition after nasal injury, hospitalized in the Department of Paediatric Otolaryngology at the Medical University of Lublin, Poland. The mean age was 11 years and 5 months (137 months) with standard deviation of 2 years and 5 months (29 months). Informed consent was obtained from the parents. The study was approved by the Bioethical Committee at the Medical University of Lublin (Resolution No. KE-0254 /216 /2012).

Mediastinal Parathyroidectomy: Preoperative Management of Hyperparathyroidism

Dariusz Sagan1*, Jerzy S. Tarach2, Andrzej Nowakowski2, Maria Klatka3, Elzbieta Czekajska-Chehab4, Andrzej Drop4, Beata Chrapko5 and Janusz Klatka6 1Department of Thoracic Surgery, 2Department of Endocrinology, 3Department of Paediatric Endocrinology and Neurology, 41st Department of Radiology, 5Department of Nuclear Medicine, 6Department of Otolaryngology and Laryngeal Oncology, Medical University of Lublin, Poland