The Relationship between Cancer Stage, Selected Immunological Parameters, Epstein-Barr Virus Infection, and Total Serum Content of Iron, Zinc, and Copper in Patients with Laryngeal Cancer.

(1) Background: the purpose of the study was to assess the relationship between cancer stage, selected immunological parameters, Epstein-Barr virus (EBV) infection, and total serum content of iron, zinc, and copper in patients with laryngeal cancer (LC). (2) Methods: serum Fe, Zn, and Cu were measured in 40 LC patients and 20 controls. Immunophenotyping of peripheral blood lymphocytes was performed by flow cytometry using fluorescent antibodies against CD3, CD4, CD8, CD19, CD25, CD69, and PD-1. Tumor and lymph node lymphocytes were analyzed by flow cytometry. EBV DNA was quantified by real-time PCR, targeting the EBNA-1 gene. Associations between serum elements, immune markers, and cancer grade/stage were evaluated using ANOVA and appropriate nonparametric tests. (3) Results: levels of Fe, Cu, and Zn were lower, while Cu/Zn was statistically higher, in patients with LC than in the control group. Correlation analysis showed a statistically significant association between the levels of these elements and parameters of the TNM (Tumor, Node, Metastasis) staging system, immunophenotype, and the amount of EBV genetic material in patients with LC who survived for more than 5 years. (4) Conclusion: the results suggest that the total serum levels of the determined micronutrients may significantly affect the immunopathogenesis and progression of LC.

The Role of Toll-like Receptor 2 (TLR2) in the Development and Progression of Hashimoto's Disease (HD): A Case Study on Female Patients in Poland.

Chronic lymphocytic thyroiditis, commonly known as HD, is one of the most common thyroid disorders. Due to the diverse factors affecting the etiopathogenesis of this disease (hormonal disorders and genetic and environmental factors), as well as the direct involvement of the immune system, scientists are increasingly willing to undertake research aimed at explaining the impact of the loss of immune tolerance and reactivity of autoantigens on the development of the disease. One of the directions of research in recent years is the role of the innate immune response, particularly Toll-like receptors (TLRs), in the pathogenesis of HD. The purpose of this study was to determine the importance of Toll-like receptor 2 (TLR2) expression on selected populations of immune cells, namely, monocytes (MONs) and dendritic cells (DCs), in the course of HD. Particular attention was paid to the analysis of TLR2's correlation with clinical parameters and the possibility its use as a potential biomarker molecule in the diagnostic process. Based on the obtained results, we found a statistically significant increase in the percentage of all analyzed populations of immune cells, i.e., mDC BDCA-1+CD19-, pDC BDCA-1+CD123, classical MONs CD14+CD16-, and non-classical MONs CD14+CD16+ showing on their surface TLR2 expression in patients diagnosed with HD compared to the healthy volunteers. Moreover, in the study group, we noted a more than 6-fold increase in the concentration of the soluble form of TLR2 in plasma compared to healthy patients. In addition, the correlation analysis showed significant positive correlations between the level of TLR2 expression on selected subpopulations of immune cells and biochemical indicators of thyroid function. Based on the obtained results, we can assume that TLR2 may be involved in the immunopathogenesis of HD.

Effect of Epstein-Barr Virus Infection on Selected Immunological Parameters in Children with Type 1 Diabetes.

Diabetes mellitus is a group of metabolic disorders with different etiologies, pathogeneses and clinical pictures, characterized by chronic hyperglycemia due to abnormal insulin secretion or action. Type 1 diabetes mellitus is the most common type of diabetes mellitus in children and adolescents, accounting for about 90% of diabetes in the population under the age of 18. The etiopathogenesis of type 1 diabetes is multifactorial. The disease occurs as a result of the interaction of three factors: genetic predisposition, environmental factors and the immune response. Research in recent years has focused on the involvement of Epstein-Barr virus (EBV) in the pathogenesis of type I diabetes. The goals of treating type 1 diabetes include maintaining blood-glucose, fructosamine and glycated hemoglobin (HbA1c) levels; therefore, the main purpose of this study was to evaluate the effect of EBV infection on the activation of selected immune cells, fructosamine levels and HbA1c levels in children with type I diabetes. Based on our study, we found a lower percentage of CD8+ T lymphocytes with expression of the CD69 molecule in patients with anti-VCA antibodies in the IgG class, and a lower percentage of CD8+ T lymphocytes with expression of the CD25+ molecule in patients with anti-EBNA-1 antibodies in the IgG class, which may indicate limited control of the immune system during EBV infection in patients. There was a lower percentage of CD3+CD4+ T lymphocytes secreting IL-4 in the study group, indicating that a deficiency in IL-4 production may be related to the development of type 1 diabetes. There was an increase in the percentage of CD4+CD3+IL-10 lymphocytes in the study group with anti-VCA antibodies present in the IgG class and anti-EBNA-1 antibodies in the IgG class compared to the patients without antibodies. In addition, there was a significant increase in fructosamine levels and higher glycated hemoglobin levels in the study group with antibodies to EBV antigens. In addition, an increase in the percentage of T lymphocytes with a CD4+CD3+IL-17+ phenotype in the patients with anti-VCA IgG antibodies was confirmed, and higher HbA1c levels may suggest that EBV infection is accompanied by an increase in IL-17 secretion.

Determinants and disparities in access to paediatricians in Poland.

BACKGROUND: The purpose of this study was to identify the factors that determine the differences in the distribution and workload of paediatricians in Poland. This research, specific to conditions found within Poland, will help further advance knowledge in this area. Data were derived from the database of Statistics Poland. The level of convergence of the phenomenon studied was analysed. The paediatricians' accessibility index was ascertained and its spatial diversity examined. The level of correlation of patients treated per paediatrician was analysed in relation to indices of urbanisation, availability of paediatricians and disposable income. RESULTS: A moderate variation of patients treated per paediatrician was found and the conditional convergence of the investigated phenomenon observed. A close negative association between the number of patients treated and access to paediatricians (-0.686, p = 0.005) was revealed. CONCLUSIONS: The research suggests that socioeconomic factors may affect the uneven spatial distribution of the workload of paediatricians in Poland and cause differences between the provinces in the equal access to paediatricians. This research may thus provide implications for policy and practice as well as lead to a better understanding of the problem.

The Clinical, Pathological, and Prognostic Value of High PD-1 Expression and the Presence of Epstein-Barr Virus Reactivation in Patients with Laryngeal Cancer.

Due to the development of molecular diagnostic techniques, the latest research in the diagnosis of cancer diseases, including laryngeal cancer, has been focused on the occurrence of specific types of molecular patterns, including markers expressed on cells of the immune system (e.g., PD-1, PD-L1, and CTLA-4), which may be directly or indirectly involved in the development of neoplastic diseases. Laryngeal cancer is one of the diseases that is diagnosed more often in men than in women, and many factors are involved in its development, including environmental and lifestyle factors, viral infections (e.g., HPV, HHV-1, and EBV), and disorders of the immune system. In this study, we determined the level of PD-1 receptor expression on T and B lymphocytes and their relationships based on the classification of the grade and TNM scale, in turn based on blood, tumor, and lymph node samples from patients diagnosed with laryngeal cancer. In addition, we determined the presence of EBV genetic material in the tested biological materials as well as the degree of cancer advancement and its correlation with the level of PD-1 receptor expression. The results suggested that the level of PD-1 expression on T and B lymphocytes was significantly higher in the tumor samples as compared to the lymph node samples, and their comparison with the immunophenotype results from the blood samples provided statistically significant data on changes in the incidence of individual subpopulations of T and B lymphocytes and the level of PD-1 receptor expression. The analysis of the individual parameters of the TNM scale also showed significant changes between the PD-1 expression and the tested biological material in individual subgroups of the scale. We also found that the expression of PD-1 on the CD4+ T cells from the lymph node samples caused an almost 1.5-fold increase in the risk of death. In the analyses of the presence of EBV, the highest concentration was recorded in the tumor samples, then for the lymph node samples, and followed by the blood samples. Furthermore, we showed that the presence of EBV genetic material was positively correlated with the level of PD-1 expression in the tested biological materials.

Vitamin D, calcium and phosphorus status in children with short stature - effect of growth hormone therapy.

OBJECTIVE: The aim of the study was to assess the level of calcium, phosphorus and vitamin D in the blood of patients treated for short stature (SS). MATERIAL AND METHODS: The study encompassed 110 children treated for somatotropin hypopituitarism (SHP) in the Department of Paediatric Endocrinology and Diabetology at the Medical University of Lublin. The levels of calcium, phosphorus and vitamin D were marked for both groups in the peripheral blood collected on a routine basis for diagnostic examinations. The parameters were compared within the group of children with SHP, both the patients who were about to start the therapy and those in the course of the therapy as well as between the research group (110 children) and the control group. RESULTS: The results obtained were compared with the results in the control group that comprised 41 children with a general good health status, although with nasal septum deviation treated in the Department of Paediatric Otolaryngology at the Medical University of Lublin. CONCLUSIONS: On the basis of the research performed, the following conclusions were drawn: 1) children with SHP were characterised with calcium-phosphorus imbalance. The level of calcium, phosphorus and vitamin D was diminished. The values did not change due to a several-year hormone growth treatment (HGT). 2) the level of calcium and phosphorus was appropriate in the control group children, but the vitamin D level was considerably lowered. This shows the necessity for vitamin D control and supplementation, not only in children with SS.

Impaired Innate Immunity in Pediatric Patients Type 1 Diabetes-Focus on Toll-like Receptors Expression.

Type 1 diabetes (DM1) is classified as an autoimmune disease. An uncontrolled response of B and T lymphocytes to the body's own tissues develops in the absence of immune tolerance. The main aim of the study was to evaluate the effect of the duration of type 1 diabetes in children on the expression of TLR receptors and the relationship with the parameters of glycemic control in patients. As a result, we showed significant differences in the level of TLR2, TLR4 and TLR9 expression in patients with DM1 in the early stage of the disease and treated chronically compared to the healthy group. Additionally, in this study, we found that the numbers of CD19+ B cells, CD3+ CD4+, CD3+ CD8+ T cells and NK cells are different for newly diagnosed DM1 individuals, patients receiving chronic treatment and for healthy controls, indicating an important role of these cells in killing pancreatic beta cells. Moreover, higher levels of IL-10 in patients with newly diagnosed DM1 have also been found, confirming the reports found in the literature.

ThymicPeptides Reverse Immune Exhaustion in Patients with Reactivated Human Alphaherpesvirus1 Infections.

Recurrent infection with human alphaherpesvirus 1 (HHV-1) may be associated with immune exhaustion that impairs virus elimination. Thymic peptides enhance immune function and thus could overcome immune exhaustion. In this study, we investigated whether reactivation of herpes infections was associated with immune exhaustion. Moreover, we examined the impact of treatment with thymostimulin on the expression of programmed cell death protein 1 (PD-1) and its ligand (PD-L1) on T and B lymphocytes in patients suffering from recurrent HHV-1 reactivation. We also assessed the effector function of peripheral blood mononuclear cells (PBMCs) after stimulation with thymic peptides. We enrolled 50 women with reactivated HHV-1 infections and healthy volunteers. We measured the expression of various activation and exhaustion markers on the surface of PBMCs using flow cytometry. In ex vivo experiments, we measured the secretion of inflammatory cytokines by PBMCs cultured with thymostimulin. Compared with controls, patients with reactivated HHV-1 infections had increased percentages of CD3+ co-expressing CD25, an activation marker (p < 0.001). Moreover, these patients had increased percentages of CD4+ and CD8+ cells co-expressing the inhibitory markers PD-1 and PD-L1. In cultures of PBMCs from the patients, thymostimulin increased the secretion of interferon gamma (p < 0.001) and interleukin (IL)-2 (p = 0.023), but not IL-4 or IL-10.Two-month thymostimulin therapy resulted in no reactivation of HHV-1 infection during this period and the reduction of PD-1 and PD-L1 expression on the surface of T and B lymphocytes (p < 0.001). In conclusion, reactivation of herpes infection is associated with immune exhaustion, which could be reversed by treatment with thymic peptides.

Socio-economic development, level of urbanization and consumption of selected food products as factors in the prevalence of overweight and obesity among youths and young adults in Poland.

INTRODUCTION AND OBJECTIVE: Obesity and overweight among teenagers and young adults pose serious problems for the Polish health care system. The aim of this study was to assess the relationships between the prevalence of overweight and obesity and the level of urbanization, consumption of selected food products and socio-economic development in the provinces in Poland. MATERIAL AND METHODS: The material used in this study is based on data published by the Central Statistics Office (GUS) for population, components of the Human Development Index (HDI), urbanization level (URBI), average monthly consumption of selected food products per person, and the percentage of obese and overweight people in the Polish provinces. The prevalence of overweight and obesity in all 16 provinces was analysed in the context of the URBI, HDI, and the average monthly consumption of selected food products. RESULTS: There was no a statistically significant correlation between the number of overweight and obesity cases and the levels of HDI and URBI. The negative correlations were found between the consumption of sugar and URBI (-0,710, p=0.002), bread and grain product consumption (-0.642; p=0.007) and fresh milk consumption (-0.594; p=0.015). Fish consumption correlated with URBI at a moderate level (0.550; p=0.027). The consumption of oils and fats strongly correlated with HDI (-0.788; p < 0,001). In the group of overweight woman, a moderate negative correlation was noted (-0.511; p=0.043) in cheese and curd consumption. CONCLUSIONS: The high level of economic development and urbanization have no direct impact on the prevalence of overweight and obesity in Poland. The results of the study indicate existing dependencies of the consumption of some of selected products with respect to the level of urbanization.

Chemical meningitis in children as a risk factor following craniopharyngioma resection - a case report.

BACKGROUND: Craniopharyngiomas are defined by the WHO as "benign" tumours, but their location and surgical treatment may be associated with major complications, one being chemical meningitis. Although rare, especially in children, it should be taken into account when worrying symptoms appear after surgery. CASE PRESENTATION: The aim of this study is to present the case of chemical meningitis in a 7-year-old girl. She was admitted to the Department of Neurology with the following symptoms: headache, vomiting and balance disorders. Brain magnetic resonance imaging showed a tumour in the sellar and suprasellar region, which was diagnosed as a craniopharyngioma. Due to acute hydrocephalus the patient underwent emergency surgery. Conventional surgery was preceded by an endocrinological consultation to determine pituitary hormone levels. The first 6 days post-surgery, during which the patient started substitution therapy for pituitary insufficiency, were uneventful but on the seventh day she presented with seizures, fever, severe headache, weakness, irritability, stiffening of the neck and a gradual degradation of consciousness. This clinical presentation suggested meningitis, which was confirmed by examination of cerebrospinal fluid. CONCLUSIONS: The conventional and/or endoscopic resection of a craniopharyngioma poses a risk of postoperative complications in the form of chemical meningitis. Although this is a rare occurrence in children with craniopharyngioma, physicians should be aware of this complication and its clinical presentation as it may facilitate earlier diagnosis, appropriate treatment and a faster recovery of their patients.

Toll-Like Receptors-2 and -4 in Graves' Disease-Key Players or Bystanders?

Graves' disease (GD) is an autoimmune disease that affects the thyroid. The development of autoimmunity is associated with innate immune responses where the prominent role plays Toll-like receptors (TLRs). The aim of our study was to assess the relationship between the expression levels of TLR-2 and TLR-4 on CD4+ and CD8+ T as well as CD19+ B lymphocytes in patients with GD and selected clinical parameters. The study group consisted of 32 women with GD, the control group consisted of 20 healthy women. Immunophenotyping was performed using the flow cytometry and cytokines concentrations were assessed using ELISA assay. The mean percentage of CD4+/TLR-2+ and CD8+/TLR-2+ T cells in patients with GD was higher than in the control group (p < 0.0001). After obtaining euthyroidism, the mean percentage of CD4+/TLR-2+ T cells in patients with GD decreased (p < 0.0001). The expression level of TLR-2 on CD4+ T lymphocytes correlated with serum FT3 concentration in patients with GD (r = 0.47, p = 0.007). The mean percentage of CD8+/TLR-2+ T cells in patients with GD before treatment compared to patients with GD after obtaining euthyroidism was higher (p = 0.0163). Similar findings were found for TLR-4. Thus the TLR-2 and TLR-4 can be a prognostic marker for Graves' disease.

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).

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

BACKGROUND: SHORT syndrome is a rare genetic congenital defects condition. The frequency of the disease still remains unknown. CASE PRESENTATION: We 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. CONCLUSIONS: The frequency of the disease still remains unknown; solely several dozen cases have been described worldwide.

Comparison of the quality of night paediatric urgent care in rural and urban areas of Lublin Province, eastern Poland - Appraisals by parents of children requiring medical attention.

 Introduction. The quality of primary medical care for children in Poland is unsatisfactory. In the ranking known as 'the European Health Consumer Index', Poland (taking the patient point of view on healthcare quality) is classified on the 27th position out of the 33 possible. The unsolved problems concern inter alia the quality and availability of night paediatric urgent care. OBJECTIVE: The aim was assessing the quality as well as the level of satisfaction with the night paediatric urgent care in the Lublin Province of eastern Poland. MATERIALS AND METHOD: The materials for this study consisted of 540 parents of children aged 6-16 years benefiting from night paediatric urgent medical assistance in Lublin Province. The survey was conducted using the Original Survey Questionnaire. RESULTS: Inhabitants of the Lublin Province (regardless of place of residence) generally assessed the quality and accessibility of night paediatric urgent care facilities as only satisfactory. Inhabitants living in rural areas have worse access to night paediatric urgent care facilities because of having to travel greater distances, and receive less comprehensive medical assistance than inhabitants living in more urbanized areas, and they are more often referred to hospital emergency departments. During the past five years, both the availability and quality of night paediatric urgent care did not change significantly. CONCLUSIONS: Inhabitants of the Lublin Province (regardless of place of residence) generally assessed the quality as well as accessibility of night paediatric urgent care facilities as only satisfactory. Rural residents have more reasons for dissatisfaction than urban dwellers. Both the quality and availability of such medical care needs to be improved.

[The coexistence of two independent disease units - Graves' disease and growth hormone deficiency in a 13 year old female patient]. / Wspólistnienie dwóch niezaleznych jednostek chorobowych ­ choroby Gravesa-Basedowa i somatotropinowej niedoczynnosci przysadki u 13-letniej pacjentki.

INTRODUCTION: Graves' disease (MGB) is the most common cause of hyperthyroidism in children and adolescents. This condition is more common in girls than boys, in a ratio of about 5:1. The coexistence of Graves' disease (MGB) with growth hormone deficiency (GHD) is not common. Much more often GHD connects with hypothyroidism. CASE REPORT: We present the case of 13-year old female patient with Graves' disease, diagnosed at 10 years of age and growth hormone deficiency, which was confirmed at the age of 12. Currently, the girl was qualified to growth hormone therapy. CONCLUSIONS: 1) Described by our case is interesting because of the rarity of coexistence Graves' disease with growth hormone deficiency. 2) GH deficiency often accompanies hypothyroidism. Note, however, that hyperthyroidism may be associated with short stature in the course of growth hormone deficiency. 3) The coexistence of endocrine disorders in childhood is not uncommon, so the diagnosis of one disease entity should not exclude further observation for the presence of other endocrine disorders.

Concentration of Selected Metals in Whole Blood, Plasma, and Urine in Short Stature and Healthy Children.

The short stature in children is defined as height below the third percentile from the mean for age and gender. This problem affects about 3% of young people. More than 20,000 children in Poland have problems with short stature. There is not much information available in the literature on the study of metals in blood, plasma, and urine in children with short stature. The study was conducted on a group of 56 short stature Polish children and 35 healthy children. The content of metals was determined using high-performance ion chromatography and inductively coupled plasma mass spectrometry methods. The study revealed significant differences between the content of selected metals in body fluids between a short stature group and healthy children. There were significant differences in the Fe, Cu, and Ni concentrations between the groups with respect to the hormonal therapy. There were no significant differences between the groups with respect to the area where the children lived. The results showed no statistically significant differences between metal concentration and age, body weight, and height. The study demonstrated statistically significant differences between the content of metals in body fluids in short stature children compared with the healthy children. It seems that the difference in the concentration of certain elements may also be the result of growth hormone therapy and the interaction between various metals. Both the alterations in the content of metals and their mutual interactions may play an important role in the pathogenesis of short stature children.

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 Spearman's 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.

[Types of cell death].

Homeostasis in the body is maintained by multiple processes, including the balance between the formation of new cells and their dying. This paper describes the mechanism and pathways of apoptotic cell death, and the phenomenon of necrosis and autophagy. Furthermore, it describes rarely presented types of cell death, namely mitotic catastrophe, oncosis and pyroptosis. The course of these processes varies, as emphasized in the paper, and is on the one hand connected with maintaining the stability of tissues in physiological processes, but on the other hand processes of cell death are connected with the induction and progression of pathological processes. In the paper an attempt was made to show the basic differences between the described processes and their importance for the body.

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.

Treatment of Graves' disease with methimazole in children alters the proliferation of Treg cells and CD3+ T lymphocytes.

Almost all cases of hyperthyroidism in children result from Graves' disease (GD). Recent studies have confirmed a significant role of T regulatory cells (Tregs) in the development of autoimmune diseases. However, the interactions between T cell responses and Treg proliferation in GD are still poorly understood. The aim of this study was to assess the proliferation of Treg cells (Tregs) and CD3+ T lymphocytes isolated from 50 children with GD before and after treatment with the thyreostatic drug methimazole (MMI). The proliferation rates, measured by methyl-3H-thymidyne incorporation, of CD3+ cells and Tregs stimulated with mitogen phorbol 12-myristate 13-acetate (PMA) were compared with those of unstimulated cells. The proliferation rates of both PMA-stimulated and unstimulated CD3+ cells prior to treatment with MMI were significantly higher than after treatment. Simultaneously, the proliferation rates of both PMA-stimulated and unstimulated Tregs were significantly lower before MMI treatment. Moreover, we observed higher cell proliferation rates of unstimulated and PMA-stimulated Tregs before the initiation of MMI therapy and after treatment in patients who had no relapse of hyperthyroidism. There was a positive correlation between the CD3+ cells proliferation rate before MMI treatment and fT3, as well as fT4 concentration in peripheral blood. The proliferation rates of CD3+ T cells before and after MMI treatment positively correlated with the TSI index. Thus, children suffering from Graves' disease presented lower Tregs proliferative potential compared with CD3+ T cells. Cocultures of CD3+ T cells and Tregs showed that Tregs were not capable of efficiently inhibiting the proliferation of CD3+ T cells in GD patients. Conclusions. MMI treatment reduced the proliferative activity of CD3+ T cells in pediatric GD patients and increased the proliferation rate of Tregs. We suggest that Treg cells that are partly dysfunctional in GD disease are probably suppressed by CD3+ T cells and that methimazole exerts some immunomodulatory effects.