Apesar das recomendações da Organização Mundial da Saúde, na reunião ministerial realizada em Amsterdam, Holanda em 2000, a tuberculose continua em ritmo crescente, atingindo principalmente os países em desenvolvimento e pessoas com o sistema imunológico comprometido, principalmente as infectadas pelo vírus da imunodeficiência adquirida. Os métodos utilizados no diagnóstico continuam os mesmos utilizados por muitos anos, cujas limitações impedem a ação rápida dos programas de saúde que buscam interromper a cadeia de transmissão da doença. Continuando uma linha de pesquisa do Laboratório de Soroepidemiologia e Imunobiologia do IMTSP, utilizando o método do Western Blotting, procuramos neste trabalho ampliar os conhecimentos da resposta imunológica de anticorpos em pacientes com tuberculose pulmonar, clinica e laboratorialmente definida, avaliando a participação das imunoglobulinas IgG, IgA e IgM, subclasses de IgG (IgG1 e IgG3) e a avidez da imunoglobulina IgG em amostras de soros colhidas no início e no final do tratamento. Nossos resultados mostraram que o melhor marcador imunológico foi a imunoglobulina IgG por apresentar melhor desempenho diagnóstico quando comparada com os resultados dos métodos microbiológicos e de dosagem de interferon gama, Quantiferon TB Gold. As frações protéicas que apresentaram melhor desempenho diagnóstico foram as de 38 e 30 KDa / Despite the recommendations of the World Health Organization, during the Ministerial meeting held in Amsterdam, Holland in 2000, tuberculosis continues at an important increasing rate, affecting mostly developing countries and people with severe compromised immune systems, especially those infected with human acquired immunodeficiency virus. The methods used for diagnosis are the same used for many years, whose limitations prevent the rapid action of countries health programs that seek to stop the chain of disease transmission. Continuing a line of research of the Laboratory of Seroepidemiology and Immunobiology of IMTSP using the method of Western blotting, in this study we tried to broaden the knowledge of the immune response of antibodies in patients with pulmonary tuberculosis, clinical and laboratory defined by evaluating the participation of IgG, IgA and IgM, IgG subclasses (IgG1 and IgG3) and immunoglobulin IgG avidity in serum samples collected in the beginning and end of treatment. Our results showed that the immunoglobulin IgG was best immunological marker when compared with the results of microbiological methods and determination of interferon gamma, QuantiFERON TB Gold. The proteins fractions that showed better diagnosis performance were 38 and 30 KDa

<p>IIn the first study, we investigated the functional implications of prolonged TLR signalling on IFN-γ mediated killing of mycobacteria by murine macrophages <i>in vitro</i>. TLR2, but not TLR4 ligation interfered with IFN-γ mediated killing of mycobacteria in macrophages. In terms of mechanisms, neither TNF nor nitric oxide (NO) production was significantly affected, and the refractoriness induced could be reversed with increasing amounts of IFN-γ In the second study, we aimed to identify immunological markers of diagnostic importance in both the respiratory tract and serum during pulmonary mycobacterial infection in mice. We found that increased levels of immunological markers in the respiratory tract, but not serum, correlated better with active mycobacterial infection in the lungs, suggesting that the immune response in the respiratory tract is more reflective of the infection status and pathology than the systemic response. Finally, we investigated the level and nature of immune responses to pulmonary mycobacterial infection in BALB/c and C57BL/6 mice, two mouse strains known to exhibit different susceptibilities to infection with several intracellular pathogens, including mycobacteria. We showed that increased susceptibility of BALB/c mice to early mycobacterial infection was associated with reduced Th1 immune responses, and increased sTNFR secretion in the lung. Moreover, BALB/c mice recruited fewer monocytes/macrophages to the lung, and although IFN-γ stimulation of infected bone marrow derived macrophages in both mouse strains resulted in induction of antimycobacterial activity, BALB/c mice had a reduced capacity to kill ingested bacteria. The work presented in this thesis provide further insight into the mechanisms involved in the host-pathogen interaction; from persistence, to the immunological processes induced by the pathogen, to susceptibility of the host to infection.</p>

IIn the first study, we investigated the functional implications of prolonged TLR signalling on IFN-γ mediated killing of mycobacteria by murine macrophages in vitro. TLR2, but not TLR4 ligation interfered with IFN-γ mediated killing of mycobacteria in macrophages. In terms of mechanisms, neither TNF nor nitric oxide (NO) production was significantly affected, and the refractoriness induced could be reversed with increasing amounts of IFN-γ In the second study, we aimed to identify immunological markers of diagnostic importance in both the respiratory tract and serum during pulmonary mycobacterial infection in mice. We found that increased levels of immunological markers in the respiratory tract, but not serum, correlated better with active mycobacterial infection in the lungs, suggesting that the immune response in the respiratory tract is more reflective of the infection status and pathology than the systemic response. Finally, we investigated the level and nature of immune responses to pulmonary mycobacterial infection in BALB/c and C57BL/6 mice, two mouse strains known to exhibit different susceptibilities to infection with several intracellular pathogens, including mycobacteria. We showed that increased susceptibility of BALB/c mice to early mycobacterial infection was associated with reduced Th1 immune responses, and increased sTNFR secretion in the lung. Moreover, BALB/c mice recruited fewer monocytes/macrophages to the lung, and although IFN-γ stimulation of infected bone marrow derived macrophages in both mouse strains resulted in induction of antimycobacterial activity, BALB/c mice had a reduced capacity to kill ingested bacteria. The work presented in this thesis provide further insight into the mechanisms involved in the host-pathogen interaction; from persistence, to the immunological processes induced by the pathogen, to susceptibility of the host to infection.

Apesar das recomendações da Organização Mundial da Saúde, na reunião ministerial realizada em Amsterdam, Holanda em 2000, a tuberculose continua em ritmo crescente, atingindo principalmente os países em desenvolvimento e pessoas com o sistema imunológico comprometido, principalmente as infectadas pelo vírus da imunodeficiência adquirida. Os métodos utilizados no diagnóstico continuam os mesmos utilizados por muitos anos, cujas limitações impedem a ação rápida dos programas de saúde que buscam interromper a cadeia de transmissão da doença. Continuando uma linha de pesquisa do Laboratório de Soroepidemiologia e Imunobiologia do IMTSP, utilizando o método do Western Blotting, procuramos neste trabalho ampliar os conhecimentos da resposta imunológica de anticorpos em pacientes com tuberculose pulmonar, clinica e laboratorialmente definida, avaliando a participação das imunoglobulinas IgG, IgA e IgM, subclasses de IgG (IgG1 e IgG3) e a avidez da imunoglobulina IgG em amostras de soros colhidas no início e no final do tratamento. Nossos resultados mostraram que o melhor marcador imunológico foi a imunoglobulina IgG por apresentar melhor desempenho diagnóstico quando comparada com os resultados dos métodos microbiológicos e de dosagem de interferon gama, Quantiferon TB Gold. As frações protéicas que apresentaram melhor desempenho diagnóstico foram as de 38 e 30 KDa / Despite the recommendations of the World Health Organization, during the Ministerial meeting held in Amsterdam, Holland in 2000, tuberculosis continues at an important increasing rate, affecting mostly developing countries and people with severe compromised immune systems, especially those infected with human acquired immunodeficiency virus. The methods used for diagnosis are the same used for many years, whose limitations prevent the rapid action of countries health programs that seek to stop the chain of disease transmission. Continuing a line of research of the Laboratory of Seroepidemiology and Immunobiology of IMTSP using the method of Western blotting, in this study we tried to broaden the knowledge of the immune response of antibodies in patients with pulmonary tuberculosis, clinical and laboratory defined by evaluating the participation of IgG, IgA and IgM, IgG subclasses (IgG1 and IgG3) and immunoglobulin IgG avidity in serum samples collected in the beginning and end of treatment. Our results showed that the immunoglobulin IgG was best immunological marker when compared with the results of microbiological methods and determination of interferon gamma, QuantiFERON TB Gold. The proteins fractions that showed better diagnosis performance were 38 and 30 KDa

Introdução: Os efeitos da radiação ultravioleta sobre o sistema imunológico humano são altamente complexos e alteram alguns componentes da resposta imunológica. Objetivo: O objetivo deste estudo foi avaliar os efeitos clínicos, histopatológicos e imunológicos da radiação solar em pescadores do sexo masculino com mais de dez anos de atividade ininterrupta. Métodos: Um estudo prospectivo, transversal, observacional e analítico, foi realizado para determinar as lesões dermatológicas diagnosticadas pelo exame físico, comparando grupos, para a análise de marcadores imunológicos na pele e no sangue, assim como alterações histológicas na pele. Este estudo foi realizado numa comunidade de pescadores, no Pina, no estado de Pernambuco, Brasil. Dezenove pescadores, com tempo médio de profissão de 29,0 ± 10,3 anos, foram incluídos no estudo. As variáveis desta amostra foram: idade, sexo, tipo da pele (segundo classificação de Fitzpatrick) estado civil, grau de instrução, número de filhos, tipo e tempo de atividade profissional, índice de massa corpórea, exposição diária à radiação solar e qualquer tipo de doença atual ou prévia (nos 12 meses anteriores à coleta de dados). As variáveis dermatológicas foram quaisquer alterações em pele, mucosa e anexos. Para comparar a subpopulação de linfócitos no sangue, foram empregados 10 indivíduos não pescadores, vivendo na mesma região e exercendo profissão ao abrigo do sol. As idades médias igualaram-se a 42,5 ± 16,1 anos. Os marcadores imunológicos da pele foram determinados por imuno-histoquímica e os do sangue, por citômetro de fluxo. O teste de Mann-Whitney, para a hipótese de igualdade, entre os grupos expostos e não expostos ao sol, foi usado. O teste de Fisher foi empregado para análise de independência dos grupos e o teste de Wilcoxon, para comparação dos achados imunológicos e histopatológicos em pele exposta e coberta, todos em igual nível de significância (0,05). Resultados: Comparando pele exposta à coberta, elastose (73,7% contra 23,1%, respectivamente; p=0,03), vasos ectásicos dérmicos (78,9% contra 31,6%, respectivamente; p=0,012) e número de células nos segmentos da epiderme entre os cones (5,8 ± 1,08 contra 5,2 ± 0,42; p=0,029) foram significantemente mais freqüentes na pele exposta. Também os marcadores CD45RO+, CD68+ e mastócitos (p=0,040, p<0,001 e p=0,001) foram estatisticamente significantes na pele exposta. O aumento de CD3CD8CD45RO+ no sangue periférico foi mais freqüente em pescadores do que em não pescadores (p=0,016). Conclusões: O efeito barreira à penetração da radiação solar, representado por elastose, aumento do número de células nas camadas entre os cones, aumento de melanócitos e da vasculatura dérmica, representada pela ectasia, sugere a existência de um efeito de tolerância ao dano da radiação solar, o qual provavelmente inibe a instalação da imunodepressão. Esse efeito é reforçado pelo aumento do CD3CD8CD45RO+ e pelo aumento da expressão da linhagem CD28+, capaz de proteger as células CD4+ da apoptose induzida pelo CD95 (Faz). SUMMARY Introduction: The ultraviolet radiation effects on the human immunological system are highly complex, disturbing some components of immune response. Objective: The objective of this study was to evaluate the clinical, histopathological and immunological effects from solar radiation in male fishermen with more than 10 years of uninterrupted activity. Methods: A prospective, transversal, observational and analytical study was done observing the dermatological lesions diagnosed by a physical exam, comparing groups, for the analysis of immunologic markers on the skin and in the blood, as well as histological alterations on the skin. This study was developed at a fishing community in Pina, in the state of Pernambuco, Brazil. Nineteen fishermen with an average professional working time of 29,0 ± 10,3 years were included in this study. The variable of this sample was: age, sex, skin type (according to Fitzpatrick classification), civilian status, degree of instruction, number of children, type and time of professional activity, body mass index, daily sun radiation exposure and any kind of current or past (12 months prior of the collection of data) illness. The dermatological variable was any alterations on the skin, mucosa and annexes. In order to compare the lymphocytes subpopulation in the blood, we used 10 non fishermen living in the same region with an indoor profession. The average ages ranged 42,5 ± 11,6 years. The immunological markers of the skin were determined by immune-histochemistry and those of blood, by flow cytometer. The Mann-Whitney test had been used, for equality hypothesis, between the sun-exposed and non-exposed group. The Fisher test was used for independence group analyzing and Wilcoxon test, as a comparison between the immunological and histopathological findings on exposed and non-exposed skin, all in equal level of significance (0,05). Results: Comparing exposed and non-exposed skin, elastosis (73,7% against 23,1% respectively; p = 0,03) dermis ectasic vases (78,9% against 31,6% respectively; p=0,012) and number of cells in epidermis segments between cones(5,8 ± 1,08 against 5,2 ± 0,42; p=0,029) were significantly more frequent in the exposed skin. Also, the CD45RO+, Cd68+ markers and mastocytes (p=0,040, p<0,001 and p=0,001) had been significantly statistic on exposed skin. The increase of CD3CD8CD45RO+, in peripheral blood, was more frequent in fishermen than the non fishermen workers (p=0,016) Conclusions: The barrier effect to the penetration of the solar radiation established by elastosis, the increase of cellular number of cell layers between the cones, increase of melanocytes and increase of dermal vasculature, represented for the ectasy, suggests the existence of a tolerance effect to sun radiation damage, which probably inhibits the installation of immunodepression. This effect is endorsed by the increasing of CD3CD8CD45RO+ and increase in trend of CD28+ expression, capable to protect Cd4+ cells apoptosis induced from CD95 (Fas). / Introduction: The ultraviolet radiation effects on the human immunological system are highly complex, disturbing some components of immune response. Objective: The objective of this study was to evaluate the clinical, histopathological and immunological effects from solar radiation in male fishermen with more than 10 years of uninterrupted activity. Methods: A prospective, transversal, observational and analytical study was done observing the dermatological lesions diagnosed by a physical exam, comparing groups, for the analysis of immunologic markers on the skin and in the blood, as well as histological alterations on the skin. This study was developed at a fishing community in Pina, in the state of Pernambuco, Brazil. Nineteen fishermen with an average professional working time of 29,0 ± 10,3 years were included in this study. The variable of this sample was: age, sex, skin type (according to Fitzpatrick classification), civilian status, degree of instruction, number of children, type and time of professional activity, body mass index, daily sun radiation exposure and any kind of current or past (12 months prior of the collection of data) illness. The dermatological variable was any alterations on the skin, mucosa and annexes. In order to compare the lymphocytes subpopulation in the blood, we used 10 non fishermen living in the same region with an indoor profession. The average ages ranged 42,5 ± 11,6 years. The immunological markers of the skin were determined by immune-histochemistry and those of blood, by flow cytometer. The Mann-Whitney test had been used, for equality hypothesis, between the sun-exposed and non-exposed group. The Fisher test was used for independence group analyzing and Wilcoxon test, as a comparison between the immunological and histopathological findings on exposed and non-exposed skin, all in equal level of significance (0,05). Results: Comparing exposed and non-exposed skin, elastosis (73,7% against 23,1% respectively; p = 0,03) dermis ectasic vases (78,9% against 31,6% respectively; p=0,012) and number of cells in epidermis segments between cones(5,8 ± 1,08 against 5,2 ± 0,42; p=0,029) were significantly more frequent in the exposed skin. Also, the CD45RO+, Cd68+ markers and mastocytes (p=0,040, p<0,001 and p=0,001) had been significantly statistic on exposed skin. The increase of CD3CD8CD45RO+, in peripheral blood, was more frequent in fishermen than the non fishermen workers (p=0,016) Conclusions: The barrier effect to the penetration of the solar radiation established by elastosis, the increase of cellular number of cell layers between the cones, increase of melanocytes and increase of dermal vasculature, represented for the ectasy, suggests the existence of a tolerance effect to sun radiation damage, which probably inhibits the installation of immunodepression. This effect is endorsed by the increasing of CD3CD8CD45RO+ and increase in trend of CD28+ expression, capable to protect Cd4+ cells apoptosis induced from CD95 (Fas).

Introdução: Os efeitos da radiação ultravioleta sobre o sistema imunológico humano são altamente complexos e alteram alguns componentes da resposta imunológica. Objetivo: O objetivo deste estudo foi avaliar os efeitos clínicos, histopatológicos e imunológicos da radiação solar em pescadores do sexo masculino com mais de dez anos de atividade ininterrupta. Métodos: Um estudo prospectivo, transversal, observacional e analítico, foi realizado para determinar as lesões dermatológicas diagnosticadas pelo exame físico, comparando grupos, para a análise de marcadores imunológicos na pele e no sangue, assim como alterações histológicas na pele. Este estudo foi realizado numa comunidade de pescadores, no Pina, no estado de Pernambuco, Brasil. Dezenove pescadores, com tempo médio de profissão de 29,0 ± 10,3 anos, foram incluídos no estudo. As variáveis desta amostra foram: idade, sexo, tipo da pele (segundo classificação de Fitzpatrick) estado civil, grau de instrução, número de filhos, tipo e tempo de atividade profissional, índice de massa corpórea, exposição diária à radiação solar e qualquer tipo de doença atual ou prévia (nos 12 meses anteriores à coleta de dados). As variáveis dermatológicas foram quaisquer alterações em pele, mucosa e anexos. Para comparar a subpopulação de linfócitos no sangue, foram empregados 10 indivíduos não pescadores, vivendo na mesma região e exercendo profissão ao abrigo do sol. As idades médias igualaram-se a 42,5 ± 16,1 anos. Os marcadores imunológicos da pele foram determinados por imuno-histoquímica e os do sangue, por citômetro de fluxo. O teste de Mann-Whitney, para a hipótese de igualdade, entre os grupos expostos e não expostos ao sol, foi usado. O teste de Fisher foi empregado para análise de independência dos grupos e o teste de Wilcoxon, para comparação dos achados imunológicos e histopatológicos em pele exposta e coberta, todos em igual nível de significância (0,05). Resultados: Comparando pele exposta à coberta, elastose (73,7% contra 23,1%, respectivamente; p=0,03), vasos ectásicos dérmicos (78,9% contra 31,6%, respectivamente; p=0,012) e número de células nos segmentos da epiderme entre os cones (5,8 ± 1,08 contra 5,2 ± 0,42; p=0,029) foram significantemente mais freqüentes na pele exposta. Também os marcadores CD45RO+, CD68+ e mastócitos (p=0,040, p<0,001 e p=0,001) foram estatisticamente significantes na pele exposta. O aumento de CD3CD8CD45RO+ no sangue periférico foi mais freqüente em pescadores do que em não pescadores (p=0,016). Conclusões: O efeito barreira à penetração da radiação solar, representado por elastose, aumento do número de células nas camadas entre os cones, aumento de melanócitos e da vasculatura dérmica, representada pela ectasia, sugere a existência de um efeito de tolerância ao dano da radiação solar, o qual provavelmente inibe a instalação da imunodepressão. Esse efeito é reforçado pelo aumento do CD3CD8CD45RO+ e pelo aumento da expressão da linhagem CD28+, capaz de proteger as células CD4+ da apoptose induzida pelo CD95 (Faz). SUMMARY Introduction: The ultraviolet radiation effects on the human immunological system are highly complex, disturbing some components of immune response. Objective: The objective of this study was to evaluate the clinical, histopathological and immunological effects from solar radiation in male fishermen with more than 10 years of uninterrupted activity. Methods: A prospective, transversal, observational and analytical study was done observing the dermatological lesions diagnosed by a physical exam, comparing groups, for the analysis of immunologic markers on the skin and in the blood, as well as histological alterations on the skin. This study was developed at a fishing community in Pina, in the state of Pernambuco, Brazil. Nineteen fishermen with an average professional working time of 29,0 ± 10,3 years were included in this study. The variable of this sample was: age, sex, skin type (according to Fitzpatrick classification), civilian status, degree of instruction, number of children, type and time of professional activity, body mass index, daily sun radiation exposure and any kind of current or past (12 months prior of the collection of data) illness. The dermatological variable was any alterations on the skin, mucosa and annexes. In order to compare the lymphocytes subpopulation in the blood, we used 10 non fishermen living in the same region with an indoor profession. The average ages ranged 42,5 ± 11,6 years. The immunological markers of the skin were determined by immune-histochemistry and those of blood, by flow cytometer. The Mann-Whitney test had been used, for equality hypothesis, between the sun-exposed and non-exposed group. The Fisher test was used for independence group analyzing and Wilcoxon test, as a comparison between the immunological and histopathological findings on exposed and non-exposed skin, all in equal level of significance (0,05). Results: Comparing exposed and non-exposed skin, elastosis (73,7% against 23,1% respectively; p = 0,03) dermis ectasic vases (78,9% against 31,6% respectively; p=0,012) and number of cells in epidermis segments between cones(5,8 ± 1,08 against 5,2 ± 0,42; p=0,029) were significantly more frequent in the exposed skin. Also, the CD45RO+, Cd68+ markers and mastocytes (p=0,040, p<0,001 and p=0,001) had been significantly statistic on exposed skin. The increase of CD3CD8CD45RO+, in peripheral blood, was more frequent in fishermen than the non fishermen workers (p=0,016) Conclusions: The barrier effect to the penetration of the solar radiation established by elastosis, the increase of cellular number of cell layers between the cones, increase of melanocytes and increase of dermal vasculature, represented for the ectasy, suggests the existence of a tolerance effect to sun radiation damage, which probably inhibits the installation of immunodepression. This effect is endorsed by the increasing of CD3CD8CD45RO+ and increase in trend of CD28+ expression, capable to protect Cd4+ cells apoptosis induced from CD95 (Fas). / Introduction: The ultraviolet radiation effects on the human immunological system are highly complex, disturbing some components of immune response. Objective: The objective of this study was to evaluate the clinical, histopathological and immunological effects from solar radiation in male fishermen with more than 10 years of uninterrupted activity. Methods: A prospective, transversal, observational and analytical study was done observing the dermatological lesions diagnosed by a physical exam, comparing groups, for the analysis of immunologic markers on the skin and in the blood, as well as histological alterations on the skin. This study was developed at a fishing community in Pina, in the state of Pernambuco, Brazil. Nineteen fishermen with an average professional working time of 29,0 ± 10,3 years were included in this study. The variable of this sample was: age, sex, skin type (according to Fitzpatrick classification), civilian status, degree of instruction, number of children, type and time of professional activity, body mass index, daily sun radiation exposure and any kind of current or past (12 months prior of the collection of data) illness. The dermatological variable was any alterations on the skin, mucosa and annexes. In order to compare the lymphocytes subpopulation in the blood, we used 10 non fishermen living in the same region with an indoor profession. The average ages ranged 42,5 ± 11,6 years. The immunological markers of the skin were determined by immune-histochemistry and those of blood, by flow cytometer. The Mann-Whitney test had been used, for equality hypothesis, between the sun-exposed and non-exposed group. The Fisher test was used for independence group analyzing and Wilcoxon test, as a comparison between the immunological and histopathological findings on exposed and non-exposed skin, all in equal level of significance (0,05). Results: Comparing exposed and non-exposed skin, elastosis (73,7% against 23,1% respectively; p = 0,03) dermis ectasic vases (78,9% against 31,6% respectively; p=0,012) and number of cells in epidermis segments between cones(5,8 ± 1,08 against 5,2 ± 0,42; p=0,029) were significantly more frequent in the exposed skin. Also, the CD45RO+, Cd68+ markers and mastocytes (p=0,040, p<0,001 and p=0,001) had been significantly statistic on exposed skin. The increase of CD3CD8CD45RO+, in peripheral blood, was more frequent in fishermen than the non fishermen workers (p=0,016) Conclusions: The barrier effect to the penetration of the solar radiation established by elastosis, the increase of cellular number of cell layers between the cones, increase of melanocytes and increase of dermal vasculature, represented for the ectasy, suggests the existence of a tolerance effect to sun radiation damage, which probably inhibits the installation of immunodepression. This effect is endorsed by the increasing of CD3CD8CD45RO+ and increase in trend of CD28+ expression, capable to protect Cd4+ cells apoptosis induced from CD95 (Fas).

INTRODUÇÃO: A alergia ao látex é um importante problema de saúde pública, especialmente em grupos de risco que têm contato frequente com este potente alérgeno. Este estudo estimou a prevalência e os fatores de risco para sensibilização ao látex em pacientes com mielomeningocele (MMC) submetidos a procedimentos cirúrgicos urológicos no HC-FMUSP. MÉTODOS: Foram selecionados pacientes com MMC submetidos a pelo menos uma cirurgia urológica, entre 2009 e 2014.Todos foram entrevistados e seus prontuários revisados. Uma amostra de sangue permitiu que a IgE específica ao látex, a K82, e seus recombinantes fossem investigados pelo método lmmunoCAP100 (kUa/L -1). A associação entre a exposição e o desfecho foi avaliada por meio de regressão logística de Poisson, Quiquadrado ou o teste exato de Fischer, para variáveis categóricas. O teste t de Student foi utilizado para comparar variáveis contínuas (nível de significância de 5%). Foram calculados a razão de prevalência (RP) e o intervalo de confiança de 95%. RESULTADOS: Foram identificados Duzentos e doze pacientes (51% do sexo masculino, 20,4 ± 6,4 anos de idade), 68 foram submetidos a pelo menos um procedimento urológico e 51 aceitaram participar (87,9%). Vinte e nove pacientes foram considerados não-sensibilizados (IgE específica para o látex :: a 0,7 kUa/L) e 22 sensibilizados ao látex com IgE > 0,7 kUa/L. Quando comparados os dois grupos, o sensibilizado apresentou um número de procedimentos cirúrgicos maior (11,6 ± 5,9 vs 7,2 ± 5,6) e dentre eles 48,3% apresentaram alguma alergia anterior contra 27,6% no grupo não sensibilizado. A sensibilização ao látex foi independentemente associada com alergia a produtos de látex (p = 0,014) e com o número de cirurgias anteriores (p = 0,032). A alergia ao látex tinha uma razão de prevalência de 2,87 (95% Cl: 1,24 a 6,65) ajustado para o número de cirurgias. Para cada procedimento cirúrgico, ajustado à alergia a produtos que contém látex, aumentou o risco para sensibilização em 4% (PR = 1,04; 95% CI: 1,00-1,09). CONCLUSÕES: A história de alergia ao látex e o número de cirurgias foram fatores de risco independentes para sensibilização ao látex / BACKGROUND: Latex allergy is an important public health issue, particularly in well defined groups who have contact with this potent allergen. This study estimated the prevalence and risk factors of latex sensitization in patients with myelomeningocele (MMC) undergoing urologic surgical procedures. METHODS: Patients with MMC that underwent at least one urologic surgery between 2009 and 2014 were interviewed and their medical records reviewed. A blood sample allowed the latex-specific lgE and its recombinant allergens by lmmunoCAP100 (kUa/L -1) to be investigated. The association between exposure and outcome was assessed by Poisson regression logistics, Chi-square or Fisher\'s exact compared for categorical variables. Student\'s t test was used to compare continuous variables (level of significance at 5%). The prevalence ratio (PR) and 95% confidence interval were calculated. RESULTS: Two hundred and twelve patients were identified (51% male, 20.4 ± 6.4 years\' age), 68 had undergone urological procedures and 51 consented to participate (87.9%). Twenty-nine patients were considered non-sensitized (latex-specific lgE :: 0,7 kUa/L -:: 1) and 22 sensitized to latex with IgE > 0,7 kUa/L. The latter group had a higher number of cirurgical procedures (11.6 ± 5.9 vs 7.2 ± 5.6) and 48.3% referred some previous allergy when compared to the non-sensitized group (27.6%). Latex sensitization was independently associated with allergy to latex products (p = 0.014) and with the number of previous surgeries (p = 0.032). Latex allergy had a prevalence ratio of 2.87 (95% Cl: 1.24 to 6.65) adjusted to the to the number of surgeries. Each surgical procedure, adjusted to allergy to latex products, increased in 4% the risk for latex sensitization (PR = 1.04; 95% Cl: 1.00 to 1.09). CONCLUSIONS: The history of latex allergy and the number of surgeries were independent risk factors for sensitization to latex

INTRODUÇÃO: A alergia ao látex é um importante problema de saúde pública, especialmente em grupos de risco que têm contato frequente com este potente alérgeno. Este estudo estimou a prevalência e os fatores de risco para sensibilização ao látex em pacientes com mielomeningocele (MMC) submetidos a procedimentos cirúrgicos urológicos no HC-FMUSP. MÉTODOS: Foram selecionados pacientes com MMC submetidos a pelo menos uma cirurgia urológica, entre 2009 e 2014.Todos foram entrevistados e seus prontuários revisados. Uma amostra de sangue permitiu que a IgE específica ao látex, a K82, e seus recombinantes fossem investigados pelo método lmmunoCAP100 (kUa/L -1). A associação entre a exposição e o desfecho foi avaliada por meio de regressão logística de Poisson, Quiquadrado ou o teste exato de Fischer, para variáveis categóricas. O teste t de Student foi utilizado para comparar variáveis contínuas (nível de significância de 5%). Foram calculados a razão de prevalência (RP) e o intervalo de confiança de 95%. RESULTADOS: Foram identificados Duzentos e doze pacientes (51% do sexo masculino, 20,4 ± 6,4 anos de idade), 68 foram submetidos a pelo menos um procedimento urológico e 51 aceitaram participar (87,9%). Vinte e nove pacientes foram considerados não-sensibilizados (IgE específica para o látex :: a 0,7 kUa/L) e 22 sensibilizados ao látex com IgE > 0,7 kUa/L. Quando comparados os dois grupos, o sensibilizado apresentou um número de procedimentos cirúrgicos maior (11,6 ± 5,9 vs 7,2 ± 5,6) e dentre eles 48,3% apresentaram alguma alergia anterior contra 27,6% no grupo não sensibilizado. A sensibilização ao látex foi independentemente associada com alergia a produtos de látex (p = 0,014) e com o número de cirurgias anteriores (p = 0,032). A alergia ao látex tinha uma razão de prevalência de 2,87 (95% Cl: 1,24 a 6,65) ajustado para o número de cirurgias. Para cada procedimento cirúrgico, ajustado à alergia a produtos que contém látex, aumentou o risco para sensibilização em 4% (PR = 1,04; 95% CI: 1,00-1,09). CONCLUSÕES: A história de alergia ao látex e o número de cirurgias foram fatores de risco independentes para sensibilização ao látex / BACKGROUND: Latex allergy is an important public health issue, particularly in well defined groups who have contact with this potent allergen. This study estimated the prevalence and risk factors of latex sensitization in patients with myelomeningocele (MMC) undergoing urologic surgical procedures. METHODS: Patients with MMC that underwent at least one urologic surgery between 2009 and 2014 were interviewed and their medical records reviewed. A blood sample allowed the latex-specific lgE and its recombinant allergens by lmmunoCAP100 (kUa/L -1) to be investigated. The association between exposure and outcome was assessed by Poisson regression logistics, Chi-square or Fisher\'s exact compared for categorical variables. Student\'s t test was used to compare continuous variables (level of significance at 5%). The prevalence ratio (PR) and 95% confidence interval were calculated. RESULTS: Two hundred and twelve patients were identified (51% male, 20.4 ± 6.4 years\' age), 68 had undergone urological procedures and 51 consented to participate (87.9%). Twenty-nine patients were considered non-sensitized (latex-specific lgE :: 0,7 kUa/L -:: 1) and 22 sensitized to latex with IgE > 0,7 kUa/L. The latter group had a higher number of cirurgical procedures (11.6 ± 5.9 vs 7.2 ± 5.6) and 48.3% referred some previous allergy when compared to the non-sensitized group (27.6%). Latex sensitization was independently associated with allergy to latex products (p = 0.014) and with the number of previous surgeries (p = 0.032). Latex allergy had a prevalence ratio of 2.87 (95% Cl: 1.24 to 6.65) adjusted to the to the number of surgeries. Each surgical procedure, adjusted to allergy to latex products, increased in 4% the risk for latex sensitization (PR = 1.04; 95% Cl: 1.00 to 1.09). CONCLUSIONS: The history of latex allergy and the number of surgeries were independent risk factors for sensitization to latex

Submitted by PPG Pediatria e Sa?de da Crian?a (pediatria-pg@pucrs.br) on 2018-05-17T17:25:42Z No. of bitstreams: 1 TESE Thiago Viola Final.pdf: 3221820 bytes, checksum: 98e6a1c6936e1a12973e6fe2d7d12ce5 (MD5) / Approved for entry into archive by Caroline Xavier (caroline.xavier@pucrs.br) on 2018-05-30T16:45:08Z (GMT) No. of bitstreams: 1 TESE Thiago Viola Final.pdf: 3221820 bytes, checksum: 98e6a1c6936e1a12973e6fe2d7d12ce5 (MD5) / Made available in DSpace on 2018-05-30T16:49:19Z (GMT). No. of bitstreams: 1 TESE Thiago Viola Final.pdf: 3221820 bytes, checksum: 98e6a1c6936e1a12973e6fe2d7d12ce5 (MD5) Previous issue date: 2018-03-12 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Introduction: Child development in adverse environments and conditions, such as with the lack of economic resources or with parental care deprivation, is considered a major risk factor for neurological and psychiatric diseases. Altered cognitive processing is thought to mediate this relationship, however, the neurobiological mechanisms underlying the effects of early adverse experiences on cognition have not yet been fully revealed. Evidence indicates that dopaminergic neurotransmission and the corticotrophinergic system have important functions in the neurobiology of decision-making and risk assessment, which are cognitive processes associated with the functionality of the cerebral cortex. Similarly, working memory is another cognitive domain that underlies cortical activity, and some studies indicate that alterations in neuroimmunologic signaling may contribute to the decline of these higher order cognitive functions. Objectives: To investigate the effects of impoverished housing conditions during early life on risk assessment processing and its associated cortical neurobiological and epigenetic mechanisms in C57BL/6 adolescent mice. In addition, we investigated the effects maternal care deprivation during early life, and the effects of systemic activation of the toll-type receptor (TLR)-3 on working memory performance, and its associated cortical neurobiological mechanisms in male BALB/c mice. Methods: Two studies with rodent experimental models were proposed. The first study used a model of impoverished housing from the postnatal day (P) 2 to P9. During adolescence, risk assessment was investigated using a behavioral paradigm that explores the conflict between two biologically relevant stimuli: the motivation to consume a sweet and highly palatable solution while being threatened by predatory olfactory cues. The expression of dopaminergic (Drd1, Drd2) and corticotrophinergic (Cfr, Crfr1) genes in the medial prefrontal cortex (mPFC) were investigated by real-time PCR. The accumulation of histone marks (H3K9me3, H3R2me2s) were assessed at the promoter region of genes associated with behavioral outcomes. In addition, plasma corticosterone levels were assessed by ELISA. In the second study, a rodent model of maternal care deprivation from P2 to P15 was applied. During adolescence, animals were injected with a TLR-3 agonist, which is a viral receptor implicated with inflammatory signaling, and then tested in a working memory task. The expression of pro-inflammatory genes (Nfkb1, Il6 and Tnf-?) and the receptor itself (Tlr3), were performed in the mPFC by real-time PCR. Results: In the first study, we found increased anxiety-like behavior, increased HPA axis response to stress and impaired RA processing in female adolescent mice, with no effect in males. These sex-specific effects were associated with increased Crfr1 mRNA expression in the medial prefrontal cortex (mPFC), which correlated with an increase in the occupancy of the histone mark H3R2me2s, a histone modification known to be involved in transcriptional activation and epigenetic priming, within the promoter of the Crfr1 gene. In the second study, we found that systemic administration of a TLR-3 agonist can modulate and exacerbate early life stress induced working memory impairments, and that higher gene expression levels of Nfkb1 in the mPFC was associated a lower working memory performance. Conclusions: The findings of the first study indicated a deleterious effect of impoverished housing exposure on risk assessment processing in females, which could be detrimental for cognitive performance in potentially dangerous situations, and suggest that the epigenetic priming of the Crfr1 gene may represent a critical factor mediating the relationship between early life stress and altered cognitive processing later in life in females. Finally, the findings of the second study demonstrated that the systemic activation of TLR-3 can induce working memory impairments, revealing an important mediating role of the neuroinflammatory signalling in the cerebral cortex associated with the cognitive changes resulting from maternal care deprivation exposure during early in life. / Introdu??o: O desenvolvimento infantil em ambientes e condi??es adversas, como frente a escassez de recursos econ?micos ou de cuidado parental, ? considerado fator de risco para doen?as neurol?gicas e psiqui?tricas. Altera??es em processos cognitivos parecem mediar esta rela??o, contudo, os mecanismos neurobiol?gicos adjacentes aos efeitos de experi?ncias adversas precoces sobre a cogni??o ainda n?o foram completamente revelados. Evid?ncias apontam que a neurotransmiss?o dopamin?rgica e o sistema corticotrofin?rgico possuem importantes fun??es na neurobiologia da tomada de decis?o e avalia??o do risco, que s?o processos cognitivos associados a funcionalidade do c?rtex cerebral. Similarmente, a mem?ria de trabalho ? outro dom?nio cognitivo que envolve atividade cortical, e alguns estudos apontam que altera??es na sinaliza??o neuroimunol?gica podem contribuir para o decl?nio destas fun??es cognitivas superiores. Objetivos: Investigar o efeito da exposi??o a moradia empobrecida na inf?ncia sobre o processamento cognitivo de avalia??o do risco e mecanismos neurobiol?gicos e epigen?ticos corticais associados em camundongos adolescentes da linhagem C57BL/6. Al?m disso, investigar o efeito da priva??o do cuidado materno na inf?ncia e da ativa??o sist?mica do receptor do tipo toll (TLR)-3 sobre a mem?ria de trabalho e mecanismos neurobiol?gicos corticais associados em camundongos machos adolescentes da linhagem BALB/c. M?todos: Foram propostos dois estudos com modelos experimentais murinos. O primeiro estudo utilizou um modelo de moradia empobrecida do dia p?s-natal (P) 2 ao P9. Quando os animais encontravam-se no per?odo da adolesc?ncia, o processamento de avalia??o do risco foi investigado por uma tarefa que explora um conflito entre dois est?mulos biologicamente fundamentais na vida de um roedor, a motiva??o de consumir uma solu??o doce e altamente palat?vel (leite condensado) tendo que se expor a pistas olfativas de um predador natural, o coiote. Os n?veis de express?o de genes dopamin?rgicos (Drd1, Drd2) e corticotrofin?rgicos (Cfr, Crfr1) no c?rtex medial pr?-frontal (mPFC) foram investigados por PCR em tempo real. Os n?veis de altera??es de histonas (H3K9me3, H3R2me2s) foram avaliados na regi?o promotora de genes associados aos desfechos comportamentais. Adicionalmente, os n?veis de corticosterona plasm?tica foram avaliados por ELISA. No segundo estudo, o modelo de adversidade utilizado foi o de priva??o do cuidado materno do P2 ao P15. Similarmente, quando os animais encontravam-se no per?odo da adolesc?ncia, ocorreu a administra??o sist?mica de um agonista de TLR-3, um receptor viral relacionado a sinaliza??o inflamat?ria, e posteriormente os animais foram testados em uma tarefa de mem?ria de trabalho. Os n?veis de express?o g?nica de genes pr?-inflamat?rios (Nfkb1, Il6 e Tnf-?) e do pr?prio receptor (Tlr3), foram avaliados no mPFC por PCR em tempo real. Resultados: no primeiro estudo, observou-se um aumento de comportamentos do tipo ansioso, maior responsividade do eixo Hipot?lamo-Pituit?ria-Adrenal (HPA) e uma diminui??o do processamento de avalia??o do risco nas f?meas expostas a moradia empobrecida, ao passo que n?o ocorreram altera??es nos animais machos. A diminui??o de avalia??o do risco foi associada a um aumento na express?o de Crfr1 no mPFC, o que se correlacionou com um aumento dos n?veis de H3R2me2s na regi?o promotora deste gene. No segundo estudo, observou-se que a ativa??o sist?mica de TLR-3 exacerbou os preju?zos de mem?ria de trabalho decorrentes da exposi??o a priva??o do cuidado materno, e este efeito correlacionou-se aos n?veis de express?o de Nfkb1 no mPFC. Conclus?es: os achados do estudo 1 indicam um efeito delet?rio da exposi??o a moradia prec?ria na inf?ncia sobre o processamento de avalia??o do risco em f?meas, revelando um preju?zo espec?fico referente ao engajamento cognitivo frente a situa??es potencialmente perigosas. Al?m disso, evidenciou-se um efeito a n?vel epigen?tico de regula??o da express?o cortical de Crfr1, indicando um importante papel deste gene sobre a rela??o entre pobreza na inf?ncia e altera??es cognitivas em f?meas adolescentes. Por fim, os achados do estudo 2 demonstraram que a ativa??o sist?mica do TLR-3 pode exacerbar os preju?zos de mem?ria de trabalho induzidos pelo estresse precoce, revelando um papel mediador da sinaliza??o neuroinflamat?ria no c?rtex cerebral relacionada as altera??es cognitivas decorrentes da exposi??o a priva??o do cuidado materno.