5 Abstract Type 1 diabetes (T1D) is a serious organ-specific autoimmune disease characterised by irreversible destruction of pancreatic β-cells by immune system. This process results in an absolute insulin deficiency. Both genetical predisposition and environmental factors influence the development of the disease. β-cell destruction is mediated by cellular components of an immune system. Proinflammatory Th1 response is considered as most pathological. Autoimmune destruction of β-cells can be identified by the detection of specific serum autoantibodies a long time before the T1D clinical onset. Currently, there is no efficient cure available to prevent or at least to delay the destructive insulitis. This diploma thesis describes the influence of synthetic diabetogenic autoantigens GAD65 and IA2 on the cytokine response of peripheral blood mononuclear cells (PBMC) obtained from T1D patients with regards to their antibody profile. The study has been carried out on patients with confirmed T1D diagnosis who tested positive for anti-GAD65 and/or anti-IA2 autoantibodies. By using flow cytometry we measured the cell type ratio in PBMC samples. The cells have been stimulated by three different concentrations of antigens and their IFNγ and IL-17 production has been detected by ELISPOT assay. In the case of both...

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-02-22T22:31:06Z No. of bitstreams: 1 KarlaSimoneCostaDeSouza_DISSERT.pdf: 1692324 bytes, checksum: 32db20c2cd32ce7a1804cd2222c5fe93 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-02-25T21:59:07Z (GMT) No. of bitstreams: 1 KarlaSimoneCostaDeSouza_DISSERT.pdf: 1692324 bytes, checksum: 32db20c2cd32ce7a1804cd2222c5fe93 (MD5) / Made available in DSpace on 2016-02-25T21:59:07Z (GMT). No. of bitstreams: 1 KarlaSimoneCostaDeSouza_DISSERT.pdf: 1692324 bytes, checksum: 32db20c2cd32ce7a1804cd2222c5fe93 (MD5) Previous issue date: 2013-07-31 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / A osteopatia ? uma complica??o cr?nica do diabetes tipo 1 (DM1). Alguns mecanismosv?m sendo propostos como principais fatores que desencadeiam altera??es no tecido ?sseo, dentreeles: a idade ao diagn?stico, tempo de doen?a, a presen?a de nefropatia e o controle glic?micoinsatisfat?rio. Neste sentido, o objetivo do presente estudo foi avaliar a express?o de RNAm dosgenes TGFB1, IGF1 e IGF1R e polimorfismos nos genes LRP5, TGFB1 e IGF1 de pacientes comDM1, e associ?-los com a presen?a de altera??es no metabolismo ?sseo. Foram estudados 100indiv?duos normoglic?micos (NG) e 101 pacientes com DM1, entre 6 e 20 anos. Os pacientesdiab?ticos foram analisados em sua totalidade (grupo DM1), e subdivididos em dois grupos, deacordo com o controle glic?mico: diab?ticos compensados (grupo DM1C) e diab?ticos n?ocompensados (grupo DM1NC). Avaliou-se o controle glic?mico (glicemia de jejum ehemoglobina glicada); a fun??o renal (ureia e creatinina s?ricas, e rela??o albumina/creatinina -RAC urin?ria); e o metabolismo ?sseo (c?lcio total e ionizado, f?sforo, atividade da fosfatasealcalina total - FAL e densidade mineral ?ssea - DMO) dos indiv?duos estudados. Tamb?m foideterminada a express?o do RNAm dos genes TGFB1, IGF1 e IGF1R e polimorfismos nos genesLRP5, TGFB1 e IGF1. A maioria dos indiv?duos com DM1 (65,3%) apresentou controleglic?mico insatisfat?rio (hemoglobina glicada >8%). Em rela??o ? fun??o renal, observou-se umaumento significativo nas concentra??es de ureia s?rica nos grupos DM1, DM1C e DM1NC e umaumento da RAC no grupo DM1NC em rela??o ao NG. No tocante aos marcadores bioqu?micosdo metabolismo ?sseo houve uma diminui??o das concentra??es s?ricas de c?lcio total nos gruposDM1, DM1C e DM1NC, e das concentra??es de c?lcio ionizado no grupo DM1 quandocomparados ao grupo NG. Tamb?m, houve um aumento significativo da atividade da FAL nogrupo DM1 em rela??o ao NG. A DMO estava significativamente diminu?da no grupo DM1quando comparado ao NG, sendo observada uma preval?ncia de 16,7% de indiv?duos diab?ticostipo 1 com baixa DMO. Na an?lise molecular, foram observadas diminui??es significativas naexpress?o dos genes TGFB1 e IGF1, e aumento significativo da express?o do gene IGF1R para ogrupo DM1 quando comparados ao NG. As frequ?ncias genot?picas e al?licas dos polimorfismosestudados foram significativas apenas para o polimorfismo do gene LRP5, demonstrando umaassocia??o deste polimorfismo com a susceptibilidade ao DM1. Por?m, n?o foram observadasassocia??es entre os polimorfismos e a osteopatia diab?tica. Estes resultados sugerem que ocontrole glic?mico insatisfat?rio, em conjunto com a presen?a de fatores de risco e altera??es emgenes envolvidos intimamente no metabolismo ?sseo, interfere na forma??o deste tecido,contribuindo para uma redu??o da DMO.

Chronic illness of a child has an influence over the whole family system. Despite this fact healthy siblings have been overlooked by the majority of research. Some foreign studies declare that this population is in higher risk of developing psychsocial problems. The purpose of this thesis is to conduct a closer study on psychosocial characteristics of the siblings of children with diabetes mellitus type 1. The main concern of the theoretical part is the review of current literature dealing with the topic of siblings of chronically ill child and their families. The conducted research included quantitative and qualitative methods exploring sibling psychosocial problems and prosocial behaviour, self-esteem, siblings attitude towards illness and family functioning. The results stress out the important relationship between sibling adjustment, self-esteem and individual and family characteristics. The results also present the main themes connected with siblings feeling of fear, sadness and anger, with applied coping strategies and perceived rewards and costs of the illness for family. Keywords: siblings, diabetes mellitus type 1, psychosocial situation

Background: Type 1 diabetes mellitus is a multifactorial disease caused by beta cell destruction of Langerhans pancreatic islets. From the genetic aspect the main predisposition lays on HLA class II genes (40 - 50%), molecules of which present exogenous peptides to CD4+ T lymphocytes. Enviromental factors play a crucial role in the etiopathogenesis of T1DM. Through epigenetic regulation (e.g. DNA methylation) the genetic and enviromental factors communicate. The level of methylation in the regulatory regions can significantly affect expression of these genes. Aims: The aim of the diploma thesis was to define methylation profile of HLA DQB1 alleles in type 1 diabetes mellitus patients and determine their expression. Methods: The genotyping of HLA class II genes (HLA-DRB1, HLA-DQA1, HLA-DQB1) was performed using sequence specific primers. DNA was treated with sodium bisulfite, regulatory region of HLA DQB1 was amplified and cloned into E.coli, strain DH5α/XL1-Blue. Positive clones were sent for sequencing and results analyzed. RNA was transcribed to cDNA by reverse transcription and the level of expression was analyzed by quantitative PCR. Results: Statistically significant differences were found in total methylation of DQB1*0201 and *0302 alleles in the B section of DQB1 gene. Difference in...

Introduction: Majority of type I diabetes (type I DM) patients do not reach satisfactory levels of compensation regardless of the advances made in the available treatment. One of the basic pillars of successful type I DM treatment is thorough education. Objective: The aim of this thesis was to describe changes in glycosylated haemoglobin (HbA1c) in patients that completed a four day long educational program. Method: The retrospective analysis evaluated 40 patients with type I DM (age 32 y.o. ± 13, HbA1c before the program 67,1 mmol/mol ± 11,75, diagnosed with DM for 12,5 years ± 7,01). HbA1c was measured before the educational program and then again in a period of 3, 6, 12 and 24 months after the program completion. The program provided the patients with classes focusing on carb counting, flexible insulin dosing, effective ways to manage hypoglycemia and physical activity. Statistical data were obtained by non-parametric tests. (Kruskal-Wallis, ANOVA-repeated measures). Results: Three months after the program completion, a significant drop in HbA1 levels could be observed (67,1 mmol/mol ± 11,75 vs. 60,2 mmol/mol ± 9,52; p=0,0093). This improvement was consistently observed after the period of 6 (59,7 mmol/mol ± 9,59; p=0,0174), 12 (56,5 mmol/mol ± 9,02; p=0,0006) and 24 (57,6 mmol/mol ± 8,43;...

Introduction: Type 1 diabetes mellitus is a chronic metabolic disease caused by autoimmune destruction of pancreatic beta cells. The theory of the disease onset is derived from study of a disease course in non-obese diabetic (NOD) mice, in which the diabetes occurs due to a dysregulation of the immune system. Experimental and clinical studies showed that the autoimmunity may be abrogated by immune intervention, which if initiated early enough may at least slow down the ongoing beta cells lost and preserve residual insulin secretion. But immune intervention alone is not sufficient to restore normoglycemia in the majority of cases. Several interventional studies showed that stimulation of proliferation and/or regeneration of beta cells are necessary to restore normoglycemia in animal models. Aim of the study: To find out, if the combination of a potent immunosuppression (murine anti-thymocyte globulin (mATG), gusperimus) together with stimulation of islet regeneration (sitagliptin) will be able to slow down or reverse the course of the disease. Another aim is to identify the mechanism by which the substances act. Material and methods: All experiments were performed in female NODShiLtJ (H2g7 ) mice. The following parameters were examined at day 0, 7, 14 and 28: blood glucose, subpopulations of...

Type 1 Diabetes (T1D) is organ-specific autoimmune disease which causes pancreatic beta cells to be irreversibly destroyed. The only possible treatment represents life-lasting insulin administration. The real trigger of destructive insulitis isn't known. T1D is a multi- factorial disease involving both external and internal factors in the disease pathogenesis. The presence of autoreactive T lymphocytes in pancreas is necessary for development of diabetes. T regulatory cells have protective function in the destructive insulitis. The aim of this diploma thesis was to study cord blood T regulatory cells and their connection to type 1 diabetes development. We tried to find the difference among T regulatory cells in mononuclear cord blood cells (CBMC) in different study groups. Samples were collected from mothers suffering from T1D, gestational diabetes. Healthy controls were tested as well. Sixty-eight samples of cord blood were included in the study among the years 2009 - 2011. Samples were divided into 3 groups (CBMC from children born to T1D mothers, mothers with gestational diabetes and healthy mothers without T1D). CBMC were ana- lysed by flow cytometry. T regulatory cells (defined as CD4+CD25+) were isolated by magnetic separation (MACS). The functional capacity of these cells was studied as well by...

Introduction: Type 1 diabetes mellitus is a chronic metabolic disease caused by autoimmune destruction of pancreatic beta cells. The theory of the disease onset is derived from study of a disease course in non-obese diabetic (NOD) mice, in which the diabetes occurs due to a dysregulation of the immune system. Experimental and clinical studies showed that the autoimmunity may be abrogated by immune intervention, which if initiated early enough may at least slow down the ongoing beta cells lost and preserve residual insulin secretion. But immune intervention alone is not sufficient to restore normoglycemia in the majority of cases. Several interventional studies showed that stimulation of proliferation and/or regeneration of beta cells are necessary to restore normoglycemia in animal models. Aim of the study: To find out, if the combination of a potent immunosuppression (murine anti-thymocyte globulin (mATG), gusperimus) together with stimulation of islet regeneration (sitagliptin) will be able to slow down or reverse the course of the disease. Another aim is to identify the mechanism by which the substances act. Material and methods: All experiments were performed in female NODShiLtJ (H2g7 ) mice. The following parameters were examined at day 0, 7, 14 and 28: blood glucose, subpopulations of...

Introduction: The diploma thesis deals with the issue of compensation of type 1 diabetes mellitus during pregnancy and its relation to the health of mother and child. It further summarizes and describes the variable factors that may have a direct effect on current glycemic variability and long- term compensation of diabetes. Aims: The theoretical part describes the characteristics of type 1 diabetes mellitus and the possibilities of therapy in pregnancy. Subsequently, factors influencing the compensation of diabetes in this period are summarized and described in more detail, especially food composition and possibilities of influencing postprandial glycemia, effects of type and intensity of physical activity and psychosocial aspects related mainly to stress from potential negative effects of glycaemia on the fetus. At the end of the theoretical part, the risks and complications for the mother and the fetus arising from long-term unsatisfactory compensation of diabetes are presented. The aim of the practical part of the thesis was to determine the extent of women's knowledge about the compensation of diabetes in pregnancy, followed by determining the effect of the extent of knowledge on long-term compensation, expressed by glycated hemoglobin. Methods: Data for the diploma thesis were obtained on the...

Abstract Background: Type 1 diabetes (T1D) is a multifactorial autoimmune disease. Its incidence in Europe is continuously rising. The highest T1D risk is associated with HLA (human leukocyte antigen) class II genes. HLA class II molecules play a key role in regulation of immune response. They contribute to the selection of T cell repertoire by presenting antigenic peptides to the CD4+ T lymphocytes. HLA class II expression is controlled by regulatory module that is situated 150 - 300 base pairs upstream of the transcription- initiation site in all HLA class II genes. Polymorphisms in this region are linked to some autoimmune diseases. There were identified several promoter alleles (named QAP) in the HLA DQA1 gene promoter region. Most of the polymorphisms appear to be conserved within haplotype. Individual QAP alleles may have a different promoter strength by which they influence expression of HLA DQA1 gene alleles. Promoter strength can be modulated by DNA methylation. Aims:Our aim was to define methylation profile of HLA DQA1 promoters and determine the mRNA expression of individual alleles of HLA DQA1 gene in T1D patients. The mRNA expression level of HLA DQA1 gene alleles was determined using quantitative PCR. Methods: 30 diabetic pacients (age range 21 to 76 years), were included in this pilot...