Genotype DQ2.5/DQ2.2 (2/2) and high celiac disease risk development

Celiac disease (CD) is a genetically determined immune-mediated disorder in which gluten immunogenic peptides are presented to CD4 T cells by HLA-DQ2.5, DQ8, DQ2.2, and their combinations. CD is considered one of the most wellcharacterized autoimmune diseases, having a described environmental factor, a well-established pathogenesis, associated genetic factors, and a well-established laboratory diagnosis, although it is still considered a difficult-to-classify disease. In the last decades, advances in laboratory diagnosis with the emergence of molecular biology techniques have allowed a specific characterization of the CD-associated genotypes and, although clinically the disease management was not modified by this factor, the follow-up of patients at risk of CD development has greatly benefited from the possibility of specifically finding the inherited genotype, and whether it represents a greater or lesser risk for developing the disease. In some populations, it is already possible to calculate the exact risk associated to the inherited genome by each individual, but the genotypes available in several countries sometimes disregard the relevance of searching beyond the genotypes DQ2.5/ DQ2.5, DQ2.5/DQ8, and DQ2.5/DQ2.2, which also present a high risk for developing the disease

Genotype DQ2.5/DQ2.2 (ββ2/ββ2) and High Celiac Disease Risk Development

Celiac disease (CD) is a genetically determined immune-mediated disorder in which gluten immunogenic peptides are presented to CD4 T cells by HLA-DQ2.5, DQ8, DQ2.2, and their combinations. CD is considered one of the most well-characterized autoimmune diseases, having a described environmental factor, a well-established pathogenesis, associated genetic factors, and a well-established laboratory diagnosis, although it is still considered a difficult-to-classify disease. In the last decades, advances in laboratory diagnosis with the emergence of molecular biology techniques have allowed a specific characterization of the CD-associated genotypes and, although clinically the disease management was not modified by this factor, the follow-up of patients at risk of CD development has greatly benefited from the possibility of specifically finding the inherited genotype, and whether it represents a greater or lesser risk for developing the disease. In some populations, it is already possible to calculate the exact risk associated to the inherited genome by each individual, but the genotypes available in several countries sometimes disregard the relevance of searching beyond the genotypes DQ2.5/DQ2.5, DQ2.5/DQ8, and DQ2.5/DQ2.2, which also present a high risk for developing the disease

Análise da expressão de genes envolvidos na interação inicial entre macrófagos murinos e Fonsecaea pedrosoi ou suas frações

Tese (Doutorado)—Universidade de Brasília, Faculdade de Medicina, Pós-Graduação em Patologia Molecular, 2011.A cromoblastomicose é uma micose crônica, supurativa e granulomatosa desenvolvida no tecido cutâneo e subcutâneo, causada por vários fungos dematiáceos sendo o Fonsecaea pedrosoi, o principal fungo isolado de pacientes acometidos por esta doença. A parede celular deste fungo é composta pelas frações F1 (ß 1 3 glicana e quitina) e F2 (a 1 3 glicana e melanina), sendo importantes estruturas de reconhecimento pelo hospedeiro. Os eventos iniciais da ativação das células da resposta imune inata após a interação com o patógeno tem despertado grande interesse. Na resposta imune ao F. pedrosoi os macrófagos têm um papel fundamental no curso da infecção, pois estão envolvidas no desenvolvimento da resposta inflamatória granulomatosa e nos mecanismos microbicidas. Para que essas células produzam intermediários reativos microbicidas é necessário que sejam ativados, no curso da cromoblastomicose, F. pedrosoi é capaz de modular a ativação dos macrófagos, tornando os apenas fungistático. Uma forma de avaliar o padrão de ativação dos macrófagos, após contato com o patógeno, é analisar a expressão de genes envolvidos no reconhecimento, adesão do fungo e na produção de citocinas. Considerando que o perfil da expressão gênica de macrófagos após a interação com F. pedrosoi até o momento não foi analisado, o objetivo deste trabalho foi definir o padrão de expressão dos genes (Itga5, Clec2, Cd14, Tlr2, Tlr4, Myd88, Nf?b, N?rf e Tnf$a ) envolvidos nas etapas iniciais de interação (30min, 1h, 3h, 6h e 24h) com o fungo ou seus componentes de parede isolados. Quando analisamos a modulação da expressão de genes pela interação entre macrófagos e os conídios, os resultados demonstram que houve o aumento no número de transcritos do gene Tlr4 e Myd88 (em 30min) e Cd14 e Tlr2 (em 6h), embora os transcritos dos genes que participam da ativação da via de sinalização do NF ?B (Nf?B, Tnf$a e N?rf) não tenham sido detectados. Quando analisamos a interação entre macrófagos e a fração F1 de F. pedrosoi, vimos que houve aumento do número de transcritos nos genes Cd14, Tlr2, Nf?B e Tnf$a (em 6h) e repressão para o gene N?rf, apontando para uma sinalização da via de NF ?B, sendo possível sugerir que a interação entre o macrófago e essa fração pode modular a ativação dessa célula. Para a interação entre macrófagos e a fração F2, o tempo mais expressivo também foi de 6h, quando houve aumento no número de transcritos dos genes Clec$2, Nf?B e Tnf$a, com repressão do gene N?rf, sugerindo que essa fração parece modular a resposta do macrófago ao fungo, através do indução de transcritos do gene Clec$2 inicialmente. Os resultados sugerem que os componentes de parede celular do fungo F. pedrosoi após interagirem com macrófagos são capazes de modular os genes destas células, aumentando a capacidade destas células de reconhecer os componentes da parede celular do fungo, com subseqüentes mudanças funcionais, que a tornam incapaz de eliminar esse fungo. _______________________________________________________________________________________ ABSTRACTThe chromoblastomycosis is a chronic, suppurative and granulomatous mycosis that affects the skin and subcutaneous tissue and is caused by various different dematiaceous fungi. Fonsecaea pedrosoi is the main fungus isolated from patients suffering from this disorder. The cell wall of this fungus is composed by two fractions – the F1 fraction (β- 1-3-glucan and chitin) and the F2 fraction (α-1-3-glucan and melanin). These structures are important in the recognition of the fungus by the host immune system. Macrophages play a key role in the initial events of the innate immune reaction to this pathogen since they are involved in the developmental of a granulomatous inflammatory response and bactericidal activity. The macrophage microbicidal activity depends on reactive intermediates that need to be activated during the course of chromoblastomycosis. However, F. Pedrosoi is capable to modulate the activation of the macrophages reducing their activity only to fungistatic. The analysis of the expression of the genes involved in the recognition and adhesion of the fungus and the production of cytokines is a way to assess the pattern of activation of macrophages after their contact with the pathogen. The gene expression profile of the macrophages after their interaction with F. pedrosoi has not been analyzed so far. Consequently the aim of the present study was to define the expression pattern of the genes (Itga5, Clec2, Cd14, Tlr2, Tlr4, Myd88, Nfκb, Nκrf and Tnf-α) involved in the initial stages of interaction of the macrophage with the fungus or the distinct components of its wall (at 30min, 1h, 3h, 6h and 24h). The analysis of the modulation genes expression caused by the interaction between macrophages and conidia disclosed an increase in the number of transcripts Tlr4 and Myd88 (at 30min) and of Cd14 and Tlr2 (at 6h), although transcripts of genes participating in the activation of the signaling pathway of NF-κB (Nfκb, Tnf-α and Nκrf) could not been detected. In addition, the analysis of the interaction between macrophages and the fraction F1 of F. pedrosoi showed an increased in transcripts of the genes Cd14, Tlr2, Nfκb and Tnf-α (at 6h) and inhibition of the gene Nκrf, pointing to a signaling pathway of NF-κB, which suggests that the interaction between this fraction and the macrophage can modulate the activation of this cell. The analysis of the interaction between macrophages and the fraction F1 of F. pedrosoi, disclosed an increased numbers of transcribed genes Cd14, Tlr2, Nfκb and Tnf-α (at 6h) and repression of the gene Nκrf, pointing to a signaling pathway of NF-κB, which might suggest that the interaction between this fraction and the macrophage can modulate the activation of this cell. In considering the interaction between macrophages and the fraction F2 the time with more expression was also at 6 hours when an increase in the number of transcripts of the genes Clec-2, Nfκb and Tnf-α was detected, with simultaneous repression of the Nκrf gene, which suggests that this fraction apparently initially modulate the macrophage response to the fungus through the induction of the gene transcript Clec-2. These results suggest that components of the fungal cell wall of F. pedrosoi, after interacting with macrophages are able to modulate the genes of these cells, increasing their capacity to recognize the components of the fungal cell wall, with subsequent functional changes, which result in their inability to eliminate the fungus

A prevalência da doença celíaca é elevada entre pacientes com epilepsia?

OBJETIVO: Determinar a prevalência da doença celíaca (DC) num grupo de pacientes epilépticos atendidos em ambulatórios de especialidade de dois hospitais gerais na cidade de Brasília (DF). MÉTODO: Amostras de soro foram colhidas de 255 pacientes epilépticos (119 crianças, 136 adultos) e de uma população formada por 4405 indivíduos (2034 crianças, 2371 adultos), atendidos no Laboratório de Análises Clínicas para exames de rotina. O diagnóstico de DC foi estabelecido por meio do teste de anticorpos antiendomísio (IgA-EMA) e por biópsia do intestino delgado. RESULTADOS: dois dos 255 pacientes epilépticos (1:127) e quinze indivíduos do grupo controle (1:293) apresentaram biópsia compatível e resultados positivos no teste IgA-EMA. CONCLUSÃO: a prevalência de DC foi 2.3 vezes maior em pacientes epilépticos do que nos controles (7.84 per 1000 versus 3.41 per 1000). Apesar deste resultado não ser estatisticamente significativo pode, no entanto ser considerado altamente sugestivo da existência de prevalência aumentada de DC entre pacientes epilépticos. ___________________________________________________________________________________________________________ ABSTRACTOBJECTIVE: To assess the prevalence of celiac disease (CD) among a group of epileptic patients attending the Epilepsy Clinics of two general hospitals in the city of Brasilia (DF), Brazil. METHOD: Serum samples were collected from 255 epileptic patients (119 children, 136 adults) originating from Epilepsy Clinics, and from a control group composed by 4405 individuals (2034 children, 2371 adults) attending the Laboratory of Clinical Analysis, for routine blood testing. The diagnosis of CD was determined by the antiendomysium antibody (IgA-EMA) test and by small intestine biopsy. RESULTS: two of the 255 epileptic patients (1:127) and fifteen subjects from the control group (1:293) tested positive for the IgA-EMA assay. CONCLUSION: the prevalence of CD was 2.3 times higher in epileptic patients than in controls (7.84 per 1000 versus 3.41 per 1000). Although still not statistically significant, this result is highly suggestive of an increased prevalence of CD among epileptic patients

Catastrophic Kawasaki disease unresponsive to IVIG in a 3-month-old infant : a diagnostic and therapeutic challenge

The present report describes the severe evolution of Kawasaki disease in a three-month-old infant. The ailment was initially atypical in its presentation, with the patient exhibiting only persistent fever in association with a progressive lethargy and maculopapular rash on the face, trunk and limbs erroneously diagnosed as roseola infantum. On the 10th day of the condition, mainly due to the unexplained persistence of fever, the infant was admitted to a local hospital. The typical features of KD appeared only on the 14th day of illness with the relapse of the maculopapular rash in association with non-purulent conjunctivitis; dry, reddish and fissured lips; tongue with reddish and hypertrophic papillae; erythema and edema of the palms and soles. During the following days, the ailment rapidly evolved to a catastrophic clinical picture characterized by generalized vasculitis, splenic infarction, pulmonary thrombosis, giant right and left coronary aneurysms, dilatation of common and internal iliac arteries and progressive ischemia of the distal third of the feet resulting in necrotic lesions of both halluces. Appropriate therapy was initiated, but repeated administration of intravenous immunoglobulin G (IVIG) followed by three days of administration of methylprednisolone did not abate the intense inflammatory activity. The remission of inflammation and regression of vascular lesions were only achieved during the following five weeks after the introduction of methotrexate associated with etanercept. The report of this case aims to draw attention to severe forms of KD that exhibit an unfavorable evolution and can be extremely refractory to the conventional therapy

hsCRP and E-Selectin as Markers of Endothelial Dysfunction in Children with Type 1 Diabetes Mellitus

Introduction: This study investigated the levels of inflammatory biomarkers in healthy children and those diagnosed with type 1 diabetes (T1DM), some of whom were affected by endothelial dysfunction (ED), characterized by increased inflammation and reduced vasodilatation. Methods: Thirty-one T1DM children showing no symptoms of vascular diseases and diagnosed by ultrasound techniques as ED-positive (T1DM-ED) or negative (T1DM), and 58 sex-age-matched healthy children were investigated for circulating levels of E-selectin, s-ICAM and s-VCAM, MMP-9, high-sensitivity C-reactive protein (hsCRP), and IL-6. Results: No differences were observed in s-ICAM, MMP-9, and IL-6 levels between case and control groups. Significantly higher levels of s-VCAM (p= 0.0001) were found in the T1DM (1359.1 ± 273 ng/mL) and T1DM-ED (1358.2 ± 112 ng/mL) groups; (control - 828.5 ± 212 ng/mL). Higher levels of E-selectin (p = 0.001) were found in the T1DM-ED group (331.2 ± 77 ng/mL); (control - 222.2 ± 74 ng/mL). The values of hsCRP were higher (p = 0.002) in the T1DM-ED group (0.36 ± 0.2 mg/L) relative to control (0.15 ± 0.1 mg/L) and T1DM (0.19 ± 0.2 mg/L). The results suggest that E-selectin and hsCRP can be useful markers of ED in children with T1DM.publishersversionpublishe

Inhibitory effects of Lactobacillus casei Shirota against both Candida auris and Candida spp. isolates that cause vulvovaginal candidiasis and are resistant to antifungals

Background Vulvovaginal candidiasis (VVC), the second leading cause of genital infection in women of reproductive age, is caused by yeasts of the genus Candida. Treatment is usually empirical and performed with azoles, which have shown increasing ineffectiveness due to resistance from these species. This therapeutic challenge has led to the search for new treatment strategies. Lactobacillus spp. produce several components with microbicidal effects, such as lactic acid. These species are the main components of a healthy vaginal microbiota and have been used as probiotics. The aim of this work was to investigate the in vitro inhibitory effects of Lactobacillus casei Shirota on both the Candida spp. that cause VVC and on C. auris. Methods The microbicidal effects of L. casei Shirota on the main VVC-causing species, C. albicans, C. tropicalis, C. norvegensis and C. parapsilosis, in addition to C. auris were investigated by counting the Colony-forming Units (CFUs) after cocultivation. The antifungal activity of lactic acid against these Candida strains was assessed using the microtiter broth dilution method to determine the minimum inhibitory concentrations (MICs). The effects of L. casei Shirota on hyphal and early biofilm formation was measured by optical microscopy. Results L. casei Shirota showed inhibitory action against all tested Candida spp., ranging from 66.9 to 95.6% inhibition depending on the species. This inhibition is possibly related to the production of lactic acid, since lactic acid has shown microbicidal action against these same Candida spp. at a concentration of 5 mg/mL, which corresponds to half of the normal physiological concentration. In addition, L. casei Shirota was able to reduce the formation of C. albicans hyphae and early biofilms, showing strong anti-Candida effects. Conclusions These results suggest that L. casei Shirota has antifungal activity against the Candida species that cause VVC. L. casei also has microbicidal action against C. auris

Oral or topical administration of L-arginine changes the expression of TGF and iNOS and results in early wounds healing

PURPOSE: To evaluate the contribution of L-arginine oral or topical rout of administration in the surgical wound healing process. METHODS: L-arginine was orally or topically administrated to mice after a laparotomy model procedure. The wounds were analyzed to evaluate the granulation tissue by HE analysis, collagen deposition, iNOS and cytokines production by immunochemisyry on wound progress. Mice used in this model were healthy, immunosupressed or diabetic and all of them were treated with different concentration of L-arginine and rout of administration. RESULTS: Suggested that groups treated with L-arginine orally or topically improved wound repair when compared with non-treatad mice. L- arginine treatment stimulated TGF-β and restricted NO production leading to a mild Th1 response and collagen deposition in injured area, when it was orally administrated. Topical administration decreased IL-8 and CCR1 expression by wound cells but did not interfere with TNF-α and IL-10 production, ratifying the decrease of inflammatory response, the oral administration however, presented a higher iNOS and TGF-β expression then. L-arginine treatment also improved the improved the wound healing in immunosupressed or diabetic mice. CONCLUSION: L-arginine administrated orally or topically can be considered an important factor in the recuperation of tissues

第949回千葉医学会例会・第19回千葉大学第三内科懇話会

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Third brazilian consensus for autoantIbodies screening in HEp-2 Cells : historical perspectve, quality control and clinical associatons

O III Consenso Brasileiro para Pesquisa de Autoanticorpos em Células HEp-2 (FAN) objetivou discutir estratégias para controlar a qualidade do ensaio, promover a atualização das associações clínicas dos diversos padrões e avaliar as difculdades de implantação do II Consenso ocorrido no ano de 2002. Métodos: Nos dias 13 e 14 de abril de 2007 participaram do encontro em Goiânia pesquisadores e especialistas de diversos centros universitários e laboratórios clínicos de diferentes regiões do Brasil, com o propósito de discutir e aprovar as recomendações que visam a melhores padronização, interpretação e utilização do ensaio pelos clínicos. Foram convidados como ouvintes representantes comerciais de diferentes empresas produtoras de insumos para realização do teste de FAN. Resultados e conclusão: Dada a heterogeneidade de microscópios e reagentes disponíveis no mercado, o III Consenso enfatizou a necessidade do controle de qualidade em ensaios de imunofuorescência indireta. Foram também feitas algumas adequações na terminologia utilizada para classifcar os diferentes padrões. Finalmente, foi realizada uma atualização das associações clínicas com fnalidade de facilitar cada vez mais o melhor uso do ensaio pelos clínicos. _________________________________________________________________________________ ABSTRACTThe Third Brazilian Consensus for Autoantibodies Screening in HEp-2 Cells (ANA) had as purpose the evaluation of diffculties in the accomplishment of the 2nd Consensus recommendations that took place in the year of 2002, the discussion of strategies for quality control of the assay and the discussion of an update of the clinical associations of the several immunofuorescent patterns. Methods: Several ANA experts from university centers and private laboratories in different areas in Brazil joined the workshop in Goiânia on 2007 April 13 and 14 with the purpose of discussing and approving the recommendations for standardization, interpretation and use of the test by physicians. Commercial representatives of different ANA slide brands were also invited as listeners to the workshop. Results and conclusion: The 3rd ANA Consensus emphasized the need for quality control in indirect immunofuorescent assays since there is a considerable heterogeneity of available microscopes and reagents. It also promoted adaptations in the previously approved terminology used to classify the different patterns and fnally updated the clinical associations of the several patterns with the purpose of providing guidance for interpretation of the assay by clinical pathologists and assistant physicians