Women in childbearing age are often affected by autoimmune diseases (AD) associated with the presence of antiphospholipid antibodies (aPL) that may influence further develop-ment of their children. The primary objective of our prospective study was to determine the presence of the following aPL: anti β2 glycoprotein I (anti-β2GPI), anticardiolipin (aCL), antiphosphatidylserine, antiphosphatidylinositol, antihospha- tidylethanolamine, antiphosphatidylglycerol, antiphosphatidic acid, antiannexin V in mothers with defined AD and their children after birth, at 6 and 12 months of life, and to compare the incidence of aPL with a control group. A secondary objective of the study was a 2-year follow-up of children born to aPL negative and aPL positive mothers with AD in order to detect the possible impact of maternal AD on the health of the offspring. In children, we analysed anthropometric data, blood cell count, cerebral and abdominal ultrasound examination, transient evoked otoacoustic emission test (TEOAE), electrocardiograph (ECG), the presence and kinetics of aPL. At the age of 2 years the Bayley Scales of Infant Development (BSID-II) were used for children's assessment of motor, language and cognitive development. 31 mothers from the total examined 82 aPL positive women with AD delivered 34 neonates...

Este trabalho teve como objetivo principal investigar o desempenho das funções executivas (FEs), a relação com a velocidade de processamento da informação e com medidas subjetivas e fisiológica do estresse em indivíduos diagnosticados com Esclerose Múltipla (EM) com incapacidade leve e moderada, avaliada pela Expanded Disability Status Scale (EDSS). O primeiro estudo corresponde a uma revisão sistemática da literatura sobre o estresse e as FEs na EM. No segundo estudo, foram apresentados os dados da comparação entre grupos no desempenho FEs e as correlações entre a cognição e a velocidade de processamento da informação. Os grupos estudados foram: controles saudáveis (n = 35); EM com EDDS 3 (n = 37); EM com EDSS entre 4-6 (n = 32). O grupo de EM com EDSS entre 4-6 apresentou pior desempenho em controle inibitório e planejamento em comparação aos demais grupos, sendo que as correlações significativas entre a velocidade de processamento da informação e o desempenho das FEs estiveram mais presentes no grupo controle e de EM com EDDS 3. No terceiro estudo, foram apresentados dados das correlações entre as FEs e medidas de estresse através da percepção e do cortisol no grupo de controles saudáveis (n = 35) e de EM (n = 69). Os dados mostraram mínimas associações entre o cortisol, especialmente, no controle inibitório e a influência de baixos níveis de cortisol na resolução de problemas⁄flexibilidade cognitiva no grupo de EM em comparação ao grupo de controles saudáveis. Concluiu-se que o desempenho das FEs pode estar associado ao nível de incapacidade medido pela EDSS de indivíduos diagnosticados com EM, sendo que a velocidade de processamento da informação parece não estar associada, de forma significativa, às FEs de indivíduos com maior incapacidade. Na presente amostra, o estresse percebido não esteve associado às FEs, porém, níveis baixos de cortisol parecem prejudicar determinados subcomponentes executivos no grupo de EM. Dados da revisão sistemática mostraram uma associação indireta entre a percepção do estresse a o desempenho das FEs após diferentes intervenções. / The main objective of this study was to investigate the performance of executive functions (EF), the relationship with information processing speed and subjective and physiological measures of stress in individuals diagnosed with Multiple Sclerosis (MS); mild and moderate disability, evaluated by Expanded Disability Status Scale (EDSS). Initially, the literature on stress and EF in MS is systematically reviewed. In the second study, we present the data of the comparison between groups in the EF performance and the correlations between the cognition and the speed of information processing. The groups studied were: healthy controls (n = 35); MS with EDDS 3 (n = 37); MS with EDSS between 4-6 (n = 32). The MS group with EDSS between 4-6 had worse performance in inhibitory control and planning in comparison to the other groups, and the significant correlations between the information processing speed and the performance of the EF were more evident in the control and MS groups with EDDS 3. In the third study, data are presented on correlations between EF and stress measures through perception and cortisol in healthy controls (n = 35) and MS (n = 69). The data showed minimal associations between cortisol, especially the influence of low cortisol levels on inhibitory control and problem-solving-cognitive flexibility in the MS group compared to the healthy controls group. We concluded that the performance of EF may be associated with the level of disability measured by the EDSS of individuals diagnosed with MS, and the speed of information processing seems not to be significantly associated to the EF of individuals with greater disability. In the present sample, perceived stress was not associated with EF, but low levels of cortisol appear to impair certain executive subcomponents in the MS group. However, data from the systematic review showed an indirect association between the perception of stress and the performance of EF after different interventions.

abstract: Type 1 diabetes (T1D) is a chronic autoimmune disease characterized by progressive autoimmune destruction of insulin-producing pancreatic β-cells. Genetic, immunological and environmental factors contribute to T1D development. The focus of this dissertation is to track the humoral immune response in T1D by profiling autoantibodies (AAbs) and anti-viral antibodies using an innovative protein array platform called Nucleic Acid Programmable Protein Array (NAPPA). AAbs provide value in identifying individuals at risk, stratifying patients with different clinical courses, improving our understanding of autoimmune destructions, identifying antigens for cellular immune response and providing candidates for prevention trials in T1D. A two-stage serological AAb screening against 6,000 human proteins was performed. A dual specificity tyrosine-phosphorylation-regulated kinase 2 (DYRK2) was validated with 36% sensitivity at 98% specificity by an orthogonal immunoassay. This is the first systematic screening for novel AAbs against large number of human proteins by protein arrays in T1D. A more comprehensive search for novel AAbs was performed using a knowledge-based approach by ELISA and a screening-based approach against 10,000 human proteins by NAPPA. Six AAbs were identified and validated with sensitivities ranged from 16% to 27% at 95% specificity. These two studies enriched the T1D “autoantigenome” and provided insights into T1D pathophysiology in an unprecedented breadth and width. The rapid rise of T1D incidence suggests the potential involvement of environmental factors including viral infections. Sero-reactivity to 646 viral antigens was assessed in new-onset T1D patients. Antibody positive rate of EBV was significantly higher in cases than controls that suggested a potential role of EBV in T1D development. A high density-NAPPA platform was demonstrated with high reproducibility and sensitivity in profiling anti-viral antibodies. This dissertation shows the power of a protein-array based immunoproteomics approach to characterize humoral immunoprofile against human and viral proteomes. The identification of novel T1D-specific AAbs and T1D-associated viruses will help to connect the nodes in T1D etiology and provide better understanding of T1D pathophysiology. / Dissertation/Thesis / Doctoral Dissertation Biological Design 2015

Este trabalho teve como objetivo principal investigar o desempenho das funções executivas (FEs), a relação com a velocidade de processamento da informação e com medidas subjetivas e fisiológica do estresse em indivíduos diagnosticados com Esclerose Múltipla (EM) com incapacidade leve e moderada, avaliada pela Expanded Disability Status Scale (EDSS). O primeiro estudo corresponde a uma revisão sistemática da literatura sobre o estresse e as FEs na EM. No segundo estudo, foram apresentados os dados da comparação entre grupos no desempenho FEs e as correlações entre a cognição e a velocidade de processamento da informação. Os grupos estudados foram: controles saudáveis (n = 35); EM com EDDS 3 (n = 37); EM com EDSS entre 4-6 (n = 32). O grupo de EM com EDSS entre 4-6 apresentou pior desempenho em controle inibitório e planejamento em comparação aos demais grupos, sendo que as correlações significativas entre a velocidade de processamento da informação e o desempenho das FEs estiveram mais presentes no grupo controle e de EM com EDDS 3. No terceiro estudo, foram apresentados dados das correlações entre as FEs e medidas de estresse através da percepção e do cortisol no grupo de controles saudáveis (n = 35) e de EM (n = 69). Os dados mostraram mínimas associações entre o cortisol, especialmente, no controle inibitório e a influência de baixos níveis de cortisol na resolução de problemas⁄flexibilidade cognitiva no grupo de EM em comparação ao grupo de controles saudáveis. Concluiu-se que o desempenho das FEs pode estar associado ao nível de incapacidade medido pela EDSS de indivíduos diagnosticados com EM, sendo que a velocidade de processamento da informação parece não estar associada, de forma significativa, às FEs de indivíduos com maior incapacidade. Na presente amostra, o estresse percebido não esteve associado às FEs, porém, níveis baixos de cortisol parecem prejudicar determinados subcomponentes executivos no grupo de EM. Dados da revisão sistemática mostraram uma associação indireta entre a percepção do estresse a o desempenho das FEs após diferentes intervenções. / The main objective of this study was to investigate the performance of executive functions (EF), the relationship with information processing speed and subjective and physiological measures of stress in individuals diagnosed with Multiple Sclerosis (MS); mild and moderate disability, evaluated by Expanded Disability Status Scale (EDSS). Initially, the literature on stress and EF in MS is systematically reviewed. In the second study, we present the data of the comparison between groups in the EF performance and the correlations between the cognition and the speed of information processing. The groups studied were: healthy controls (n = 35); MS with EDDS 3 (n = 37); MS with EDSS between 4-6 (n = 32). The MS group with EDSS between 4-6 had worse performance in inhibitory control and planning in comparison to the other groups, and the significant correlations between the information processing speed and the performance of the EF were more evident in the control and MS groups with EDDS 3. In the third study, data are presented on correlations between EF and stress measures through perception and cortisol in healthy controls (n = 35) and MS (n = 69). The data showed minimal associations between cortisol, especially the influence of low cortisol levels on inhibitory control and problem-solving-cognitive flexibility in the MS group compared to the healthy controls group. We concluded that the performance of EF may be associated with the level of disability measured by the EDSS of individuals diagnosed with MS, and the speed of information processing seems not to be significantly associated to the EF of individuals with greater disability. In the present sample, perceived stress was not associated with EF, but low levels of cortisol appear to impair certain executive subcomponents in the MS group. However, data from the systematic review showed an indirect association between the perception of stress and the performance of EF after different interventions.

Dysregulation between pro- and anti-inflammatory cytokines activity in rheumatoid arthritis (RA) contributes to immune dysregulation, chronic inflammation and subsequent joint destruction. Interleukin-37 (IL-37) has been described as an anti-inflammatory cytokine in several autoimmune diseases. The main aim of this work was to determine the levels of IL-37 in serum and synovial fluid (SF) of RA patients and to compare them with the levels in patients with osteoarthritis (OA) and further explore the association of IL-37 with disease activity and other clinical parameters. Subsequent goal was to study its anti-inflammatory function on RA synovial fibroblasts and describe other cells types of synovial tissue contributing to its production. IL-37 levels were detected using enzyme-linked immunosorbent assay (ELISA). Synovial fibroblasts were stimulated by lipopolysaccharide (LPS) and recombinant IL-37 (rIL-37). The levels of studied genes were detected by PCR. Synovial tissues and immune cells were visualized by immunohistochemical and by immunofluorescence staining. We found increased levels of IL-37 in SF of patients with RA in comparison to OA patients. There was a significant correlation between serum and SF levels of IL-37. RA as well as OA patients showed increased levels of IL-37 in serum than in...

Introduction: HLA class II system presents one of the most important mechanism in immune system, which is able to recognise pathogens and damaged cells. Some HLA class II alleles are associated with autoimmune diseases, for example celiac disease, which is typical by chronic inflammation of small intestine and other following symptoms. The risk HLA class II variants are DQ2 and DQ8. Epigenetic mechanisms that regulates gene expression, especially methylation of cytosine in promoter region of DQ2 and/or DQ8 alleles, could have influence on development of T lymphocytes in the thymus, where T-lymphocytes develop and pass a few stages in, and only the survival clones can be part of function immune system. Aim: The aim of this study is to compare methylation level of promoter regions of HLA DQ2 and DQ8 alleles between celiac patients and healthy controls. Another goal is to compare expression level of DQ2 and DQ8 variants between these two groups. Methods: DNA and RNA were isolated from full blood of two sets of donors. DNA was converted by bisulphite conversion and then amplified by Nested PCR. The PCR product was cloned to bacteria. Than positive colonies were selected. Subsequent methylation analysis was performed. RNA was converted to cDNA by Reverse transcription. Relative expression was analyzed...

Este trabalho teve como objetivo principal investigar o desempenho das funções executivas (FEs), a relação com a velocidade de processamento da informação e com medidas subjetivas e fisiológica do estresse em indivíduos diagnosticados com Esclerose Múltipla (EM) com incapacidade leve e moderada, avaliada pela Expanded Disability Status Scale (EDSS). O primeiro estudo corresponde a uma revisão sistemática da literatura sobre o estresse e as FEs na EM. No segundo estudo, foram apresentados os dados da comparação entre grupos no desempenho FEs e as correlações entre a cognição e a velocidade de processamento da informação. Os grupos estudados foram: controles saudáveis (n = 35); EM com EDDS 3 (n = 37); EM com EDSS entre 4-6 (n = 32). O grupo de EM com EDSS entre 4-6 apresentou pior desempenho em controle inibitório e planejamento em comparação aos demais grupos, sendo que as correlações significativas entre a velocidade de processamento da informação e o desempenho das FEs estiveram mais presentes no grupo controle e de EM com EDDS 3. No terceiro estudo, foram apresentados dados das correlações entre as FEs e medidas de estresse através da percepção e do cortisol no grupo de controles saudáveis (n = 35) e de EM (n = 69). Os dados mostraram mínimas associações entre o cortisol, especialmente, no controle inibitório e a influência de baixos níveis de cortisol na resolução de problemas⁄flexibilidade cognitiva no grupo de EM em comparação ao grupo de controles saudáveis. Concluiu-se que o desempenho das FEs pode estar associado ao nível de incapacidade medido pela EDSS de indivíduos diagnosticados com EM, sendo que a velocidade de processamento da informação parece não estar associada, de forma significativa, às FEs de indivíduos com maior incapacidade. Na presente amostra, o estresse percebido não esteve associado às FEs, porém, níveis baixos de cortisol parecem prejudicar determinados subcomponentes executivos no grupo de EM. Dados da revisão sistemática mostraram uma associação indireta entre a percepção do estresse a o desempenho das FEs após diferentes intervenções. / The main objective of this study was to investigate the performance of executive functions (EF), the relationship with information processing speed and subjective and physiological measures of stress in individuals diagnosed with Multiple Sclerosis (MS); mild and moderate disability, evaluated by Expanded Disability Status Scale (EDSS). Initially, the literature on stress and EF in MS is systematically reviewed. In the second study, we present the data of the comparison between groups in the EF performance and the correlations between the cognition and the speed of information processing. The groups studied were: healthy controls (n = 35); MS with EDDS 3 (n = 37); MS with EDSS between 4-6 (n = 32). The MS group with EDSS between 4-6 had worse performance in inhibitory control and planning in comparison to the other groups, and the significant correlations between the information processing speed and the performance of the EF were more evident in the control and MS groups with EDDS 3. In the third study, data are presented on correlations between EF and stress measures through perception and cortisol in healthy controls (n = 35) and MS (n = 69). The data showed minimal associations between cortisol, especially the influence of low cortisol levels on inhibitory control and problem-solving-cognitive flexibility in the MS group compared to the healthy controls group. We concluded that the performance of EF may be associated with the level of disability measured by the EDSS of individuals diagnosed with MS, and the speed of information processing seems not to be significantly associated to the EF of individuals with greater disability. In the present sample, perceived stress was not associated with EF, but low levels of cortisol appear to impair certain executive subcomponents in the MS group. However, data from the systematic review showed an indirect association between the perception of stress and the performance of EF after different interventions.

京都大学 / 新制・課程博士 / 博士(生命科学) / 甲第23335号 / 生博第453号 / 新制||生||60(附属図書館) / 京都大学大学院生命科学研究科統合生命科学専攻 / (主査)教授 野田 岳志, 教授 杉田 昌彦, 教授 千坂 修 / 学位規則第4条第1項該当 / Doctor of Philosophy in Life Sciences / Kyoto University / DFAM

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Multiple sclerosis (MS) is among the most common neuroinflammatory diseases across the globe and is autoimmune mediated in nature. This progressive, highly debilitating disease often leaves individuals wheelchair bound within 15-25 years of onset. MS is characterized by inflammatory lesions that appear in unpredictable locations around the central nervous system. Lesions can be visualized using magnetic resonance imaging (MRI) technology. As neuroinflammation continues and lesions accumulate, patients can experience a wide array of progressively worsening symptoms including but not limited to motor impairments, sensory disturbances, loss of control of bodily functions, and/or neuropathic pain, depending on the location of lesion formations. There are different types of MS, the most common being relapsing-remitting MS (RRMS) seen in about 85% of cases and characterized by periods of symptom remission followed by flare-ups. A large majority of these patients go on to develop secondary progressive MS (SPMS) where neurological damage and patient decline is progressive and continuous. Primary progressive MS (PPMS) is seen in about 10% of cases and is characterized by progressive and continuous patient decline from the outset of disease. Other rarer forms of MS do exist but will not be discussed further. Research aimed at MS is at an all-time high and the timing could not be better: its global incidence and prevalence is climbing. For decades MS has been thought of as a disease caused by dysfunctional CD4+ T-helper 1 (Th1) cells. It is now known that many different cell types contribute to MS pathophysiology. These other cell types include macrophages and dendritic cells of the innate immune system due to their expression of MHC class II molecules that function to activate CD4+ Th1 cells. More recent research has implicated CD8+ T-cells and B-cells in contributing to disease through direct destruction of neural cells that express MHC class I molecules and through the generation of autoantibodies, respectively. While these discoveries are important and provide hope for future breakthrough treatments, there are still enormous gaps in the medical community’s knowledge of what causes MS. The epidemiologic pattern of MS prevalence has for many decades interested scientists and hinted at a potential cause of this disease. MS tends to affect white individuals with genetic ties to northern Europe, but this relationship may not still hold true, as MS incidence and prevalence may be rising faster in black populations compared to other races/ethnicities, at least in the United States. MS occurs nearly 3 times as often in females than in males, and is strongly associated with Epstein-Barr virus (EBV) infection—especially in those that go on to develop infectious mononucleosis (IM). MS prevalence increases markedly in regions north of 40 degrees North latitude or south of 40 degrees South latitude. MS risk also changes depending on body mass index (BMI) considerations, migration history, and in families with a genetic history of the disease. It is well-accepted that MS has a genetic component, the most important of which is the presence of the HLA-DRB1*1501 allele that codes for certain proteins in MHC class II molecules. However, genetics alone are unable to sufficiently account for MS risk as the concordance rate for identical twins with MS is only 25-30%. These well-established findings imply that some unknown environmental factor(s) must be contributing to MS initiation and progression. All of the environmental factors listed above have a common connecting thread that is logically and empirically verifiable: vitamin D. This fat-soluble vitamin can either be endogenously synthesized in the skin after exposure to ultraviolet B (UVB) light or consumed through the diet, the former being of more importance to humans. Epidemiologic patterns suggest a protective role for vitamin D in MS, where low or deficient levels of vitamin D may be a contributor to increased risk for MS. Populations living at greater latitudes, north or south, have significantly greater prevalence of MS which coincides with the reduction of endogenously produced vitamin D in these regions due to a lesser amount of UVB light (and of lower intensity) experienced year-round. Increases in BMI, especially increased adiposity, correlate with increased risk for MS and with prevalence of vitamin D deficiency. Women tend to naturally have greater adiposity than men, thus increasing their risk for MS. Estrogens and vitamin D have been shown to act synergistically to protect against MS, therefore vitamin D deficiency may increase risk for MS in women. Vitamin D is a known immunomodulatory agent that promotes tolerogenic immune states. Vitamin D also offsets many of the harmful effects caused by EBV, among these including repression of B-cell differentiation into plasma cells, reduced MHC II expression, and promotion of B cell apoptosis. This serves to repress deleterious immunoglobulin secretion by B-cells. Vitamin D is also immunologically beneficial as it promotes regulatory T cell function and their expression of protective cytokines, and through its inhibition of inflammatory Th1 and Th17 cell functions. In total, the immunomodulatory mechanisms of vitamin D are important as the immunological states produced by vitamin D are exactly the opposite of those observed in MS patients and MS animal models. Research in vitamin D is gaining attention as the scientific community is quickly discovering that its true physiologic role extends far beyond its classical function as a calcium regulator. Indeed, rapidly evolving research is revealing roles for vitamin D in cardiovascular function and blood pressure regulation, brain development and neurological function, and even in the prevention of certain cancers. However, this thesis will focus on its most well-known function secondary to calcium regulation: immunomodulation and its anti-inflammatory capabilities. The last portion of this thesis will present information advocating for the increase in minimum dietary intake of vitamin D from its current value of 800 IU/day to 5,000 IU/day. While a more than 5-fold increase may seem drastic, the tolerable upper limit is at least 10,000 IU/day even by the most conservative of estimates—the true upper limit is probably around 20,000 IU/day and may even be 50,000 IU/day. The global prevalence of vitamin D deficiency is so extensive that some authors have even considered it a global pandemic: upwards of 50% of the entire world population may be deficient in this crucial vitamin. Increasing vitamin D supplementation is an extremely low risk way to reduce risk for MS and other diseases.