Dual effect of Thymosin α 1 on human monocyte-derived dendritic cellin vitrostimulated with viral and bacterial toll-like receptor agonists

OBJECTIVES: Thymosin α 1 (Tα1) recently gained interest as immune adjuvant for vaccines because of its ability to modulate the T-cell/dendritic cell (DC) axis and to improve antibody production. The objective of this study was to determine whether Tα1 would address in vitro the response of human primary monocyte-derived DC, crucial regulators of vaccine-induced immunity, upon exposure to different toll-like receptor (TLR) agonists or infection with viruses or bacteria. METHODS: DC maturation and production of pro-inflammatory cytokines were analyzed. RESULTS: Our data revealed a dual effect of Tα1 on DC biology upon viral or bacterial stimulation. Interestingly, Tα1 enhanced human leukocyte antigen (HLA)-I and II surface expression and secretion of IL-6, TNF-α and IL-8 when DCs were treated with viral TLR3 and TLR7/8 agonists. Similarly, in pandemic H1N1 influenza A-infected DCs, Tα1 raised the expression of maturation markers and type I and III Interferon (IFN). In contrast, following bacterial TLR2 and 4 stimulation, as well as upon Bacillus Calmette-Guerin infection, the presence of Tα1 in DC cultures drastically lowered the analyzed cellular parameters. CONCLUSION: The knowledge that Tα1 pleiotropic effect might ameliorate anti-viral immune responses and, at the same time, dampen inflammation caused by bacterial infections could lay the groundwork for a more appropriate therapeutic application of this molecule

Altered CSF Albumin Quotient Links Peripheral Inflammation and Brain Damage in MS

OBJECTIVE CNS damage can increase the susceptibility of the blood-brain barrier (BBB) to changes induced by systemic inflammation. The aim of this study is to better understand BBB permeability in patients with MS and to examine whether compromised BBB integrity in some of these patients is associated with CNS damage and systemic inflammation. METHODS Routine CSF measurements of 121 patients with MS were analyzed including number and type of infiltrating cells, total protein, lactate, and oligoclonal bands, as well as intrathecal production of immunoglobulins and CSF/serum quotients for albumin, immunoglobulins, and glucose. In addition, in a subcohort of these patients, we performed ex vivo immunophenotyping of CSF-infiltrating and paired circulating lymphocytes using a panel of 13 monoclonal antibodies, we quantified intrathecal neurofilament light chain (NF-L) and chitinase 3-like 1 (CHI3L1), and we performed intrathecal lipidomic analysis. RESULTS Patients with MS with abnormal high levels of albumin in the CSF showed a distinct CSF cell infiltrate and markers of CNS damage such as increased intrathecal levels of NF-L and CHI3L1 as well as a distinct CSF lipidomic profile. In addition, these patients showed higher numbers of circulating proinflammatory Th1 and Th1* cells compatible with systemic inflammation. Of interest, the abnormally high levels of albumin in the CSF of those patients were preserved over time. CONCLUSIONS Our results support the hypothesis that CNS damage may increase BBB vulnerability to systemic inflammation in a subset of patients and thus contribute to disease heterogeneity

Confiabilidade do questionário Lista de Atividades Físicas em crianças

OBJETIVO: Evaluar la confiabilidad del cuestionario Lista de Actividades Físicas en niños. MÉTODOS: El estudio es parte de la adaptación transcultural del cuestionario, realizado con 83 escolares de siete a diez años, matriculados del 2º al 5º año de enseñanza fundamental de la ciudad de Sao Paulo, Sureste Brasil, en 2008. El cuestionario fue respondido por el niño por medio de entrevista individual, presenta lista con 21 actividades físicas moderadas a vigorosas realizadas en el día anterior, es dividido en períodos (antes, durante y posterior a la escuela) y posee sección de evaluación de la entrevista. El cuestionario permite cuantificar: tiempo en actividades físicas y sedentarias; y costos metabólicos total y ponderado. La confiabilidad fue evaluada comparándose dos entrevistas realizadas con intervalo promedio de tres horas. Para la Sección C (evaluación de la entrevista), se compararon datos de la primera entrevista y de un evaluador externo. Se utilizaron propuesta de Bland y Altman y los coeficientes de correlación intraclase y de correlación de concordancia de Lin en la evaluación de la confiabilidad. RESULTADOS: Los límites inferiores de los coeficientes de correlación intraclase para los resultados analizados variaron de 0,84 a 0,96. La precisión y depuración variaron, respectivamente, de 0,83 a 0,97 y de 0,99 a 1. La recta estimada a partir de pares de valores obtenidos en las dos aplicaciones para actividad física indica elevada precisión de los datos. El ítem de la entrevista con peor resultado fue la habilidad en estimar tiempo (regular en 27,6% de las entrevistas). Los itens de la Sección C presentaron coeficientes de correlación intraclase entre 0,60 y 0,70, excepto el nivel de cooperación (0,46). CONCLUSIONES: La Lista de Actividades Físicas presenta alta confiabilidad para contrastar actividad física y sedentaria del día anterior en niños.OBJETIVO: Avaliar a confiabilidade do questionário Lista de Atividades Físicas em crianças. MÉTODOS: O estudo é parte da adaptação transcultural do questionário, realizado com 83 escolares de sete a dez anos, matriculados do segundo ao quinto ano do ensino fundamental da cidade de São Paulo, SP, em 2008. O questionário foi respondido pela criança por meio de entrevista individual, apresenta lista com 21 atividades físicas moderadas a vigorosas realizadas no dia anterior, é dividido em períodos (antes, durante e após a escola) e possui seção de avaliação da entrevista. O questionário permite quantificar: tempo em atividades físicas e sedentárias; e custos metabólicos total e ponderado. A confiabilidade foi avaliada comparando-se duas entrevistas realizadas com intervalo médio de três horas. Para a seção C (avaliação da entrevista), compararam-se dados da primeira entrevista e de um avaliador externo. Utilizaram-se a proposta de Bland & Altman e os coeficientes de correlação intraclasse e de correlação de concordância de Lin na avaliação da confiabilidade. RESULTADOS: Os limites inferiores dos coeficientes de correlação intraclasse para os desfechos analisados variaram de 0,84 a 0,96. A precisão e concordância variaram, respectivamente, de 0,83 a 0,97 e de 0,99 a 1. A reta estimada a partir de pares de valores obtidos nas duas aplicações para atividade física indica elevada precisão dos dados. O item da entrevista com pior resultado foi a habilidade em estimar tempo (regular em 27,7% das entrevistas). Os itens da seção C apresentaram coeficientes de correlação intraclasse entre 0,60 e 0,70, exceto o nível de cooperação (0,46). CONCLUSÕES: A Lista de Atividades Físicas apresenta alta confiabilidade para aferir atividade física e sedentária do dia anterior em crianças.OBJECTIVE: To assess the reliability of the Lista de Atividades Físicas (Brazilian version of the Physical Activity Checklist Interview) in children. METHODS: The study is part of a cross-cultural adaptation of the Physical Activity Checklist Interview, conducted with 83 school children aged between seven and ten years, enrolled between the 2nd and 5th grades of primary education in the city of São Paulo, Southeastern Brazil, in 2008. The questionnaire was responded by children through individual interviews. It is comprised of a list of 21 moderate to vigorous physical activities performed on the previous day, it is divided into periods (before, during and after school) and it has a section for interview assessment. This questionnaire enables the quantification of time spent in physical and sedentary activities and the total and weighed metabolic costs. Reliability was assessed by comparing two interviews conducted with a mean interval of three hours. For the interview assessment, data from the first interview and those from an external evaluator were compared. Bland-Altman's proposal, the intraclass correlation coefficient and Lin's concordance correlation coefficient were used to assess reliability. RESULTS: The intraclass correlation coefficient lower limits for the outcomes analyzed varied from 0.84 to 0.96. Precision and agreement varied between 0.83 and 0.97 and between 0.99 and 1, respectively. The line estimated from the pairs of values obtained in both interviews indicates high data precision. The interview item showing the poorest result was the ability to estimate time (fair in 27.7% of interviews). Interview assessment items showed intraclass correlation coefficients between 0.60 and 0.70, except for level of cooperation (0.46). CONCLUSIONS: The Brazilian version of the Physical Activity Checklist Interview shows high reliability to assess physical and sedentary activity on the previous day in children

Confiabilidade do questionário Lista de Atividades Físicas em crianças

OBJETIVO: Evaluar la confiabilidad del cuestionario Lista de Actividades Físicas en niños. MÉTODOS: El estudio es parte de la adaptación transcultural del cuestionario, realizado con 83 escolares de siete a diez años, matriculados del 2º al 5º año de enseñanza fundamental de la ciudad de Sao Paulo, Sureste Brasil, en 2008. El cuestionario fue respondido por el niño por medio de entrevista individual, presenta lista con 21 actividades físicas moderadas a vigorosas realizadas en el día anterior, es dividido en períodos (antes, durante y posterior a la escuela) y posee sección de evaluación de la entrevista. El cuestionario permite cuantificar: tiempo en actividades físicas y sedentarias; y costos metabólicos total y ponderado. La confiabilidad fue evaluada comparándose dos entrevistas realizadas con intervalo promedio de tres horas. Para la Sección C (evaluación de la entrevista), se compararon datos de la primera entrevista y de un evaluador externo. Se utilizaron propuesta de Bland y Altman y los coeficientes de correlación intraclase y de correlación de concordancia de Lin en la evaluación de la confiabilidad. RESULTADOS: Los límites inferiores de los coeficientes de correlación intraclase para los resultados analizados variaron de 0,84 a 0,96. La precisión y depuración variaron, respectivamente, de 0,83 a 0,97 y de 0,99 a 1. La recta estimada a partir de pares de valores obtenidos en las dos aplicaciones para actividad física indica elevada precisión de los datos. El ítem de la entrevista con peor resultado fue la habilidad en estimar tiempo (regular en 27,6% de las entrevistas). Los itens de la Sección C presentaron coeficientes de correlación intraclase entre 0,60 y 0,70, excepto el nivel de cooperación (0,46). CONCLUSIONES: La Lista de Actividades Físicas presenta alta confiabilidad para contrastar actividad física y sedentaria del día anterior en niños.OBJETIVO: Avaliar a confiabilidade do questionário Lista de Atividades Físicas em crianças. MÉTODOS: O estudo é parte da adaptação transcultural do questionário, realizado com 83 escolares de sete a dez anos, matriculados do segundo ao quinto ano do ensino fundamental da cidade de São Paulo, SP, em 2008. O questionário foi respondido pela criança por meio de entrevista individual, apresenta lista com 21 atividades físicas moderadas a vigorosas realizadas no dia anterior, é dividido em períodos (antes, durante e após a escola) e possui seção de avaliação da entrevista. O questionário permite quantificar: tempo em atividades físicas e sedentárias; e custos metabólicos total e ponderado. A confiabilidade foi avaliada comparando-se duas entrevistas realizadas com intervalo médio de três horas. Para a seção C (avaliação da entrevista), compararam-se dados da primeira entrevista e de um avaliador externo. Utilizaram-se a proposta de Bland & Altman e os coeficientes de correlação intraclasse e de correlação de concordância de Lin na avaliação da confiabilidade. RESULTADOS: Os limites inferiores dos coeficientes de correlação intraclasse para os desfechos analisados variaram de 0,84 a 0,96. A precisão e concordância variaram, respectivamente, de 0,83 a 0,97 e de 0,99 a 1. A reta estimada a partir de pares de valores obtidos nas duas aplicações para atividade física indica elevada precisão dos dados. O item da entrevista com pior resultado foi a habilidade em estimar tempo (regular em 27,7% das entrevistas). Os itens da seção C apresentaram coeficientes de correlação intraclasse entre 0,60 e 0,70, exceto o nível de cooperação (0,46). CONCLUSÕES: A Lista de Atividades Físicas apresenta alta confiabilidade para aferir atividade física e sedentária do dia anterior em crianças.OBJECTIVE: To assess the reliability of the Lista de Atividades Físicas (Brazilian version of the Physical Activity Checklist Interview) in children. METHODS: The study is part of a cross-cultural adaptation of the Physical Activity Checklist Interview, conducted with 83 school children aged between seven and ten years, enrolled between the 2nd and 5th grades of primary education in the city of São Paulo, Southeastern Brazil, in 2008. The questionnaire was responded by children through individual interviews. It is comprised of a list of 21 moderate to vigorous physical activities performed on the previous day, it is divided into periods (before, during and after school) and it has a section for interview assessment. This questionnaire enables the quantification of time spent in physical and sedentary activities and the total and weighed metabolic costs. Reliability was assessed by comparing two interviews conducted with a mean interval of three hours. For the interview assessment, data from the first interview and those from an external evaluator were compared. Bland-Altman's proposal, the intraclass correlation coefficient and Lin's concordance correlation coefficient were used to assess reliability. RESULTS: The intraclass correlation coefficient lower limits for the outcomes analyzed varied from 0.84 to 0.96. Precision and agreement varied between 0.83 and 0.97 and between 0.99 and 1, respectively. The line estimated from the pairs of values obtained in both interviews indicates high data precision. The interview item showing the poorest result was the ability to estimate time (fair in 27.7% of interviews). Interview assessment items showed intraclass correlation coefficients between 0.60 and 0.70, except for level of cooperation (0.46). CONCLUSIONS: The Brazilian version of the Physical Activity Checklist Interview shows high reliability to assess physical and sedentary activity on the previous day in children

Brain Citrullination Patterns and T Cell Reactivity of Cerebrospinal Fluid-Derived CD4+ T Cells in Multiple Sclerosis

Immune responses to citrullinated peptides have been described in autoimmune diseases like rheumatoid arthritis (RA) and multiple sclerosis (MS). We investigated the post-translational modification (PTM), arginine to citrulline, in brain tissue of MS patients and controls (C) by proteomics and subsequently the cellular immune response of cerebrospinal fluid (CSF)-infiltrating T cells to citrullinated and unmodified peptides of myelin basic protein (MBP). Using specifically adapted tissue extraction- and combined data interpretation protocols we could establish a map of citrullinated proteins by identifying more than 80 proteins with two or more citrullinated peptides in human brain tissue. We report many of them for the first time. For the already described citrullinated proteins MBP, GFAP, and vimentin, we could identify additional citrullinated sites. The number of modified proteins in MS white matter was higher than control tissue. Citrullinated peptides are considered neoepitopes that may trigger autoreactivity. We used newly identified epitopes and previously reported immunodominant myelin peptides in their citrullinated and non-citrullinated form to address the recognition of CSF-infiltrating CD4+ T cells from 22 MS patients by measuring proliferation and IFN-γ secretion. We did not detect marked responses to citrullinated peptides, but slightly more strongly to the non-modified version. Based on these data, we conclude that citrullination does not appear to be an important activating factor of a T cell response, but could be the consequence of an immune- or inflammatory response. Our approach allowed us to perform a deep proteome analysis and opens new technical possibilities to analyze complex PTM patterns on minute quantities of rare tissue samples

Characterization of Antigen-Induced CD4+ T-Cell Senescence in Multiple Sclerosis

Antigen-induced T-cell exhaustion and T-cell senescence are peripheral regulatory mechanisms that control effector T-cell responses. Markers of exhaustion and senescence on T Cells indicate the previous activation by repetitive stimulation with specific antigens. Malignant tumors are accompanied by enhanced T-cell exhaustion and T-cell senescence resulting in immune evasion, while these control mechanisms might be diminished in autoimmune diseases including multiple sclerosis (MS). To better understand the involvement of antigen-induced T-cell senescence in controlling CD4+ T-cell-mediated autoimmune responses in MS, we have analyzed the re-expression of CD45RA and the downregulation of CD28 and CD27 molecules as markers of antigen-induced T-cell senescence in fresh cerebrospinal fluid (CSF)-infiltrating and paired circulating T cells from patients with MS. Patients with different levels of CD4+ T-cell senescence were identified and characterized regarding demographical and clinical features as well as intrathecal markers of neurodegeneration. CD4+ T-cell senescence was also analyzed in control patients to explore a putative deficit of this regulatory mechanism in MS. This study shows heterogeneity of markers of CD4+ T-cell senescence in patients with MS. Patients with high levels of CD4+ T-cell senescence in peripheral blood showed increased frequencies of CSF-infiltrating CD28+ CD27-EM CD4+ T cells with a proinflammatory Th1 functional phenotype. The correlation of these cells with the intrathecal levels of neurofilament light chain, a marker of neurodegeneration, suggests their relevance in disease pathogenesis and the involvement of T-cell senescence in their regulation. Markers of antigen-induced T-senescence, therefore, show promise as a tool to identify pathogenic CD4+ T cells in patients with MS

Dozrávání rajčat z pozdních sběrů

In this diploma thesis "Ripening tomatoes from the late harvest" were investigated supporting options of the post harvest of the completely green fruit, which remained unharvested in the later stages of harvest. Fruits were transferred from the field conditions to the temperature of 20 °C and afterwards were successively treated with ethylene as the supportive maturing factor, which helped them to get to the suitable conditions for subsequent maturation. Two varieties of tomatoes were used for the speed of ripening. At the end of the cultivation period, both varieties of tomatoes 'Expres' and 'Premio' were treated by the three concentrations of exogenous ethylene, 2 100 ppm, 7 360 ppm and 2 200 ppm. Technologically important parameters such as the size and weight of fruit, fruit color, fruit firmness, soluble solids and titration acid were evaluated during the post-harvest storage (21 days). Influence of ethylene in concentrations up to 7 360 ppm has proven effective post-harvest treatment that supports ripening of originally unripe fruit at the end of the growing season. Also differences in the investigated varieties were proved, the 'Premio' variety was after the ethylene treatment sensitive in parameters, which were monitored

Confiabilidad del Cuestionario Lista de Actividades Físicas en niños

OBJETIVO: Avaliar a confiabilidade do questionário Lista de Atividades Físicas em crianças. MÉTODOS: O estudo é parte da adaptação transcultural do questionário, realizado com 83 escolares de sete a dez anos, matriculados do segundo ao quinto ano do ensino fundamental da cidade de São Paulo, SP, em 2008. O questionário foi respondido pela criança por meio de entrevista individual, apresenta lista com 21 atividades físicas moderadas a vigorosas realizadas no dia anterior, é dividido em períodos (antes, durante e após a escola) e possui seção de avaliação da entrevista. O questionário permite quantificar: tempo em atividades físicas e sedentárias; e custos metabólicos total e ponderado. A confiabilidade foi avaliada comparando-se duas entrevistas realizadas com intervalo médio de três horas. Para a seção C (avaliação da entrevista), compararam-se dados da primeira entrevista e de um avaliador externo. Utilizaram-se a proposta de Bland & Altman e os coeficientes de correlação intraclasse e de correlação de concordância de Lin na avaliação da confiabilidade. RESULTADOS: Os limites inferiores dos coeficientes de correlação intraclasse para os desfechos analisados variaram de 0,84 a 0,96. A precisão e concordância variaram, respectivamente, de 0,83 a 0,97 e de 0,99 a 1. A reta estimada a partir de pares de valores obtidos nas duas aplicações para atividade física indica elevada precisão dos dados. O item da entrevista com pior resultado foi a habilidade em estimar tempo (regular em 27,7% das entrevistas). Os itens da seção C apresentaram coeficientes de correlação intraclasse entre 0,60 e 0,70, exceto o nível de cooperação (0,46). CONCLUSÕES: A Lista de Atividades Físicas apresenta alta confiabilidade para aferir atividade física e sedentária do dia anterior em crianças.OBJECTIVE: To assess the reliability of the Lista de Atividades Físicas (Brazilian version of the Physical Activity Checklist Interview) in children. METHODS: The study is part of a cross-cultural adaptation of the Physical Activity Checklist Interview, conducted with 83 school children aged between seven and ten years, enrolled between the 2nd and 5th grades of primary education in the city of São Paulo, Southeastern Brazil, in 2008. The questionnaire was responded by children through individual interviews. It is comprised of a list of 21 moderate to vigorous physical activities performed on the previous day, it is divided into periods (before, during and after school) and it has a section for interview assessment. This questionnaire enables the quantification of time spent in physical and sedentary activities and the total and weighed metabolic costs. Reliability was assessed by comparing two interviews conducted with a mean interval of three hours. For the interview assessment, data from the first interview and those from an external evaluator were compared. Bland-Altman's proposal, the intraclass correlation coefficient and Lin's concordance correlation coefficient were used to assess reliability. RESULTS: The intraclass correlation coefficient lower limits for the outcomes analyzed varied from 0.84 to 0.96. Precision and agreement varied between 0.83 and 0.97 and between 0.99 and 1, respectively. The line estimated from the pairs of values obtained in both interviews indicates high data precision. The interview item showing the poorest result was the ability to estimate time (fair in 27.7% of interviews). Interview assessment items showed intraclass correlation coefficients between 0.60 and 0.70, except for level of cooperation (0.46). CONCLUSIONS: The Brazilian version of the Physical Activity Checklist Interview shows high reliability to assess physical and sedentary activity on the previous day in children.OBJETIVO: Evaluar la confiabilidad del cuestionario Lista de Actividades Físicas en niños. MÉTODOS: El estudio es parte de la adaptación transcultural del cuestionario, realizado con 83 escolares de siete a diez años, matriculados del 2º al 5º año de enseñanza fundamental de la ciudad de Sao Paulo, Sureste Brasil, en 2008. El cuestionario fue respondido por el niño por medio de entrevista individual, presenta lista con 21 actividades físicas moderadas a vigorosas realizadas en el día anterior, es dividido en períodos (antes, durante y posterior a la escuela) y posee sección de evaluación de la entrevista. El cuestionario permite cuantificar: tiempo en actividades físicas y sedentarias; y costos metabólicos total y ponderado. La confiabilidad fue evaluada comparándose dos entrevistas realizadas con intervalo promedio de tres horas. Para la Sección C (evaluación de la entrevista), se compararon datos de la primera entrevista y de un evaluador externo. Se utilizaron propuesta de Bland y Altman y los coeficientes de correlación intraclase y de correlación de concordancia de Lin en la evaluación de la confiabilidad. RESULTADOS: Los límites inferiores de los coeficientes de correlación intraclase para los resultados analizados variaron de 0,84 a 0,96. La precisión y depuración variaron, respectivamente, de 0,83 a 0,97 y de 0,99 a 1. La recta estimada a partir de pares de valores obtenidos en las dos aplicaciones para actividad física indica elevada precisión de los datos. El ítem de la entrevista con peor resultado fue la habilidad en estimar tiempo (regular en 27,6% de las entrevistas). Los itens de la Sección C presentaron coeficientes de correlación intraclase entre 0,60 y 0,70, excepto el nivel de cooperación (0,46). CONCLUSIONES: La Lista de Actividades Físicas presenta alta confiabilidad para contrastar actividad física y sedentaria del día anterior en niños.Programa de Incentivo à Pesquisa da Universidade de São Paulo (USP