Prevalencia de anticuerpos anti-Saccharomyces cerevisiae en pacientes celíacos / Prevalence of anti-Saccharomyces cerevisiae antibodies in celiac patients / Prevalência de anticorpos anti-Saccharomyces cerevisiae em pacientes celíacos

La enfermedad celíaca (EC) es una condición inflamatoria crónica del intestino delgado causada por intolerancia al gluten. El tratamiento consiste en la dieta libre de gluten (DLG). Los anticuerpos anti Saccharomyces cerevisiae (ASCA) están dirigidos contra la pared celular de la levadura, se asocian a enfermedades autoinmunes, y se propone la permeabilidad intestinal alterada como causa de activación de la inmunidad humoral. El objetivo del trabajo fue determinar la prevalencia de ASCA IgG e IgA en pacientes celíacos bajo tratamiento y evaluar la asociación de ASCA con el grado de adherencia a la DLG. Se analizaron 59 sueros de pacientes adultos celíacos con alta o baja adherencia a la DLG, y se determinó ASCA IgG e IgA. Se halló una prevalencia de ASCA IgG y/o IgA del 44%. Se encontró asociación entre ASCA-IgG y adherencia a DLG (OR 4,04 IC 95%: 1,32-12,38). La prevalencia de ASCA en la población celíaca estudiada es similar a la reportada en la bibliografía. La menor prevalencia de ASCA IgG en pacientes con una estricta DLG respecto de aquellos con baja adherencia, indicaría que su presencia depende del nivel de ingesta de gluten, sugiriéndolos como herramienta complementaria en el seguimiento del paciente celíaco.

Celiac disease (CD) is a chronic inflammatory condition of the small intestine caused by gluten intolerance. The treatment consists of gluten free diet (GFD). Anti Saccharomyces cerevisiae antibodies (ASCA) are directed against the cell wall of yeast, associated with autoimmune diseases, and an altered intestinal permeability is proposed as a cause of activation of humoral immunity. The objective of this work was to determine the prevalence of IgG and IgA ASCA in celiac patients under treatment and to evaluate the association of ASCA with the degree of adherence to GFD. Fifty-nine serum samples from adult celiac patients with high or low adherence to GFD were analyzed, determining IgG and IgA ASCA. A 44% prevalence of IgG and/or IgA ASCA was found. An association was discovered between IgG ASCA and GFD adherence (OR 4.04, 95% CI: 1.32-12.38). The prevalence of ASCA in the studied celiac population is similar to that reported in the literature. The lower prevalence of IgG ASCA in patients with a strict GFD compared to those with low adherence would indicate that their presence depends on the level of gluten intake, suggesting them as a complementary tool in the follow-up of the celiac patient.

A doença celíaca (DC) é uma condição inflamatória crônica do intestino delgado causada pela intolerância ao glúten. O tratamento consiste na dieta sem glúten (DSG). Os anticorpos anti Saccharomyces cerevisiae (ASCA) são dirigidos contra a parede celular da levedura, associados a doenças autoimunes, e à permeabilidade intestinal alterada como causa da ativação da imunidade humoral. O objetivo foi determinar a prevalência de ASCA IgG e IgA em pacientes celíacos em tratamento; avaliar a associação de ASCA com o grau de adesão ao DSG. Foram analisados 59 soros de pacientes celíacos adultos com alta ou baixa adesão ao DSG, determinando ASCA IgG e IgA. Foi encontrada uma prevalência de SCA IgG/ou IgA de 44%. Foi encontrada uma associação entre ASCA-IgG e a adesão ao DSG (OR 4,04 IC 95% 1,32-12,38). A prevalência de ASCA na população celíaca estudada é semelhante à relatada na literatura. A menor prevalência de ASCA IgG em pacientes com rigorosa DSG, em comparação àqueles com baixa adesão, indicaria que sua presença depende do nível de ingestão de glúten, sugerindo-os como uma ferramenta complementar no seguimento do paciente celíaco.

A Case of Small Bowel Diffuse Large B-cell Lymphoma Mimicking Crohn's Disease / 대한소화기학회지

A 66-year-old male with dyspepsia and weight loss was referred to our hospital for evaluation. On laboratory examination, anti-saccharomyces cerevisiae (ASCA)-IgA was positive and iron deficiency anemia was present. PET/CT and abdominal CT scan images showed multiple small bowel segmental wall thickening and inflammation. Capsule endoscopy images showed multiple small bowel ulcerative lesions with exudates. Based on laboratory test results and imaging studies, the patient was diagnosed with Crohn's disease and treated with prednisolone and 5-aminosalicylic acid (5-ASA). However, the patient underwent second operation due to small bowel perforation within 2 month after initiation of treatment. Pathology report of the resected specimen was compatible to primary small bowel diffuse large B cell lymphoma and pertinent treatment was given to the patient after recovery. Herein, we describe a case of primary small bowel diffuse large B cell lymphoma that was mistaken for Crohn's disease.

Diagnostic value of ASCA and atypical p-ANCA in differential diagnosis of inflammatory bowel disease

Worldwide, the incidence of inflammatory bowel disease [IBD] is increasing. This study aims to evaluate the diagnostic value of two serological markers, atypical perinuclear anti-neutrophil cytoplasmic antibodies [atypical-P-ANCA] and anti-Saccharomyces cerevisiae antibodies [ASCA], with the intent to determine their relationship to ulcerative colitis [UC] and Crohn's disease [CD], in addition to the location and extent of bowel involvement. There were 97 patients enrolled in this study, 72 diagnosed with UC and 25 with CD. The control group consisted of 40 healthy individuals. ASCA was determined by enzyme-linked immunosorbent assay [ELISA] and atypical-P-ANCA by indirect immunofluorescence assay [IIF]. For data analyses, we used the chi-square and independent t-tests. Significance was considered to be p<0.05. For CD, the sensitivity of ASCA was 16% and its specificity was 97%. ASCA had a specifity of 90% in UC patients. The atypical P-ANCA test had a sensitivity of 44% and specificity of 86% for UC. The positive predictive value [PPV] for atypical P-ANCA in UC patients was 78% and for the negative predictive value [NPV], it was 58%.There was no correlation between ASCA and atypical P-ANCA results and the location of gastrointestinal [GI] involvement in CD [p=0.61] and UC [p=0.28] patients. According to the results, ASCA and atypical P-ANCA markers are not useful for IBD screening. Our study suggests that atypical P-ANCA is a useful parameter to differentiate UC from CD. However, ASCA is of limited value for screening and differentiating UC from CD

¿Son útiles los marcadores serológicos ASCA y p-ANCA en la enfermedad inflamatoria intestinal? / Are useful the ASCA and p-ANCA serologic markers in inflammatory bowel disease?

Background: Several studies have suggested that Anti-Saccharomyces cerevisiae antibodies (ASCA) and perinuclear anti-neutrophil cytoplasmic antibodies (p-ANCA) are useful serological markers associated with inflammatory bowel disease (IBD). However, neither the indication nor its use in clinical practice have been clearly established. Aim: to assess whether the presence of these markers have a possible diagnostic role and clinical significance. Patients and Methods: Retrospective chart review of 93 patients, average age 42 years, 48 female. ASCA and p-ANCA were determinated by ELISA and IFI. The sensitivity (S), specificity (E), positive and negative predictive values (PPV and NPV), and X2 were determinated. Results: Sixty eight patients with IBD (35 Crohn´s disease (CD), 31 ulcerative colitis (UC), one IBD unclassified and one indeterminate colitis patients) and 25 patients with other gastrointestinal diseases. In the total group of patients the S and E of ASCA and p-ANCA for diagnosis of CD and UC was 48.6 percent, 74.1 percent and 77.4 percent, 82.3 percent respectively. In patients with IBD, the presence of ASCA(+)/p-ANCA(-) had a S, E, PPV, and NPV for diagnosis of CD 37.1 percent, 93.5 percent, 86.7 percent and 56.9 percent respectively. On the other hand, the presence of ASCA(-)/p-ANCA(+) had a S, E, PPV, and NPV for diagnosis of UC 64.5 percent, 85.7 percent, 80 percent and 73.1 percent respectively. The evolution of IBD patients was not associated with the presence of these markers. Conclusions: Our study showed that both p-ANCA and ASCA did not have an important role in the differential diagnosis of CD and UC and in their prognosis. New strategies to differentiate CD and UC and to determinate their prognosis are needed.

Existen estudios que han sugerido que los anticuerpos Anti-Saccharomyces cerevisiae (ASCA) y los anticuerpos anticitoplasma de los neutrófilos perinuclear (p-ANCA), son marcadores serológicos asociados a las enfermedades inflamatorias intestinales (EII). Sin embargo, su indicación y uso en la práctica clínica no han sido aún clarificados. Objetivos: Evaluar si la presencia de estos marcadores posee algún papel en el diagnóstico y pronóstico. Pacientes y Métodos: Noventa y tres pacientes, edad promedio 42 años, 48 mujeres. Los anticuerpos ASCA fueron determinados por técnica de ELISA y los p-ANCA por IFI. Se calculó la sensibilidad (S), especificidad (E), valor predictivo positivo (VPP) y negativo (VPN) y X2. Resultados: Se incluyen sesenta y siete pacientes con EII (35 enfermedad de Crohn (EC), 31 colitis ulcerosa (CU), uno con EII no clasificable y un paciente con Colitis Indeterminada) y 25 pacientes con otras enfermedades gastrointestinales. En el grupo total de pacientes, la S y E de ASCA y p-ANCA para el diagnóstico de EC y CU fue de 48,6 y 74,1 por ciento y 77,4 y 82,3 por ciento respectivamente. En pacientes con diagnóstico establecido de EII, la presencia de ASCA(+)/p-ANCA(-) tuvo una S, E, VPP y VPN para el diagnóstico de EC 37,1, 93,5, 86,7 y 56,9 por ciento, respectivamente. Por otro lado, la presencia de ASCA(-)/p-ANCA(+) tuvo una S, E, VPP y VPN para el diagnóstico de CU 64,5, 85,7, 80 y 73,1 por ciento, respectivamente. La evolución favorable o desfavorable de los pacientes con EII (EC o CU) no se correlacionó con la presencia (positividad) de uno o ambos marcadores (p ≥ 1). Conclusiones: Nuestro estudio demostró que los marcadores serológicos ASCA y ANCA utilizados en conjunto no poseen actualmente un papel importante en la diferenciación de la EC de la CU, como tampoco para establecer un pronóstico de su evolución. Por lo tanto, es necesario encontrar nuevas estrategias para poder diferenciar estos dos cuadros y poder determinar...

Serum Anti-Saccharomyces Cerevisiae Antibodies in Greek Patients with Behcet's Disease

We tested 59 Greek patients with Behcet's Disease (BD) for serum anti-Saccharomyces cerevisiae antibodies. No increase of these antibodies was detected in the cases compared to 55 healthy unrelated blood donors from the same population. This finding is in contrast with the correlation between Saccharomyces cerevisiae antibodies and BD as reported in other populations. It seems that environmental factors may contribute to disease expression in different populations, producing different effects according to the individual's genetic predisposition. Saccharomyces cerevisiae antibodies do not seem to be of any significance in the Greek population.

Antibodies anti-Saccharomyces cerevisiae (ASCA) do not differentiate Crohn's disease from celiac disease / Anticorpos anti-Saccharomyces cerevisiae não diferenciam doença de Crohn de doença celíaca

CONTEXT: Anti-Saccharomyces cerevisiae antibodies (ASCA), considered serologic markers for Crohn's disease, were described in patients with celiac disease, disappearing after a gluten-free diet. OBJECTIVES: Evaluation of ASCA positivity in patients with Crohn's disease and celiac disease in relation to healthy individuals. METHODS: A total of 145 individuals were studied: 36 with Crohn's disease and 52 with celiac disease, that fulfilled the diagnostic criteria for both affections, and 57 healthy individuals for control. The celiac patients were divided as follow: group CeD I at diagnosis (n = 34), group CeD II with gluten-free diet compliance (n = 13) and group CeD III with transgressions to the diet (n = 5). ASCA IgA and IgG were determined by ELISA. RESULTS: With statistical significance, ASCA IgA were positive in Crohn's disease, celiac disease at diagnosis and celiac disease with diet transgressions; ASCA IgG in Crohn's disease and in all groups with celiac disease. CONCLUSIONS: The detection of ASCA in patients with celiac disease allows to suggest that ASCA is not a specific marker for Crohn's disease, but was associated with the inflammation of the small intestine. The increased levels of positive ASCA may be due to genetic factors and increased intestinal permeability.

RACIONAL: Anticorpos anti-Saccharomyces cerevisiae antibodies, considerados marcadores sorológicos para a doença de Crohn, foram descritos em pacientes com doença celíaca, desaparecendo após dieta isenta de glúten. OBJETIVOS: Avaliação da positividade de anti-Saccharomyces cerevisiae antibodies em pacientes com doença de Crohn e doença celíaca, em relação a indivíduos sadios da mesma área geográfica. MÉTODOS: Foram estudados 145 pacientes, 36 com doença de Crohn e 52 com doença celíaca que preencheram os critérios diagnósticos para ambas as afecções, e 57 indivíduos sadios para controle. Os pacientes celíacos foram divididos como segue: ao diagnóstico (grupo doença celíaca I: n = 34), obedientes à dieta isenta de glúten (grupo doença celíaca II: n = 13) e não-aderentes à dieta isenta de glúten (grupo doença celíaca III: n = 5). Anti-Saccharomyces cerevisiae antibodies IgA e IgG foram determinados por ELISA. RESULTADOS: Anti-Saccharomyces cerevisiae antibodies IgA foi positivo na doença de Crohn, nos celíacos ao diagnóstico e nos transgressores à dieta, com significado estatístico. Anti-Saccharomyces cerevisiae antibodies IgG foi positivo na doença de Crohn e em todos os grupos de celíacos, com significado estatístico. CONCLUSÕES: A detecção de anti-Saccharomyces cerevisiae antibodies em pacientes com doença celíaca permite sugerir que o mesmo não seja marcador específico para a doença de Crohn, mas que esteja associado à inflamação do intestino delgado. A positividade de anti-Saccharomyces cerevisiae antibodies pode ser decorrente de fatores genéticos e aumento da permeabilidade intestinal.

Efecto de adición de iones hierro y zinc sobre la producción de etanol de dos cepas recombinantes de Saccharomyces cerevisiae / The effect of adding iron and zinc ions to ethanol production from two recombinant Saccharomyces cerevisiae strains

La producción de etanol por fermentación es influenciada por la presencia de iones metálicos como hierro y zinc dado que son cofactores de la enzima alcohol deshidrogenasa. El estudio de este efecto permitiría identificar el comportamiento de los microorganismos fermentadores en sustratos industriales que contienen altas concentraciones de este tipo de iones. Este trabajo evaluó la producción de biomasa, los azúcares residuales y la producción de etanol por fermentación de tres cepas de S. cerevisiae, CBS8066, recombinantes GG570-CIBI y GG570-CIBII, bajo el efecto de la adición de hierro a 0, 50 y 150 M, y zinc a 0 y 50 M. Las cepas presentaron inhibición en la producción de biomasa y etanol bajo efecto de iones de hierro y zinc, siendo dicha inhibición mayor al estar en presencia de zinc o alta concentración de hierro. GG570-CIBI mostró disminución en producción de biomasa de 4 g/L y una caída en producción de etanol de 40% en el tratamiento 150 M hierro-50 M zinc (con respecto al tratamiento basal). GG570-CIBII fue la menos afectada con inhibición en la producción de etanol inferior a 11% a las 20 h de fermentación. Adicionalmente, presentó la mayor producción de etanol cuando hubo adición de 150 M Fe con o sin adición de zinc, siendo dicha producción entre un 9 y 14% superior a la de las cepas CBS8066 y GG570-CIBI respectivamente, bajo las mismas condiciones. Posteriormente, GG570-CIBII será evaluada en sustratos industriales debido a su menor inhibición en la producción de etanol, permitiendo así obtener mejores rendimientos.

The ethanol production by fermentation is influenced by the presence of metallic ions like iron and zinc because these are alcohol dehydrogenase enzyme cofactors. The study of this effect would allow for identifying the behavior of microorganisms in industrial substrates that contain high concentrations of this kind of ions. This work evaluated biomass production, residual sugars and ethanol production by fermentation of three S. cerevisiae strains, CBS8066, recombinants GG570-CIBI and GG570-CIBII, under the effect of the addition of ferrous ion at 0, 50 and 150 M and zinc ion at 0 and 50 M. The strains showed inhibition on biomass and ethanol production under the effect of zinc and ferrous ions, however, this inhibition was greater in the presence of zinc or iron at high concentration. GG570-CIBI showed reduction in biomass production of 4 g/L and an ethanol production drop of 40 % in the treatment 150 M iron–50 M zinc (with respect to the basal treatment). GG570-CIBII was the less affected with an inhibition on ethanol production below 11 % at 20 h of fermentation. Additionally, GG570-CIBII presented the greatest ethanol production when 150 M iron was added to the culture medium with or without zinc addition. In this case, the production was 9 and 14 % greater than ethanol production of CBS8066 and GG570-CIBI respectively, at the same conditions. Later, GG570-CIBII will be evaluated in industrial substrates due to its lower ethanol production inhibition, allowing for obtaining better yields.

Nitric oxide synthase activity in tissues of the blowfly Chrysomya megacephala (Fabricius, 1794)

Although insects lack the adaptive immune response of the mammalians, they manifest effective innate immune responses, which include both cellular and humoral components. Cellular responses are mediated by hemocytes, and humoral responses include the activation of proteolytic cascades that initiate many events, including NO production. In mammals, nitric oxide synthases (NOSs) are also present in the endothelium, the brain, the adrenal glands, and the platelets. Studies on the distribution of NO-producing systems in invertebrates have revealed functional similarities between NOS in this group and vertebrates. We attempted to localize NOS activity in tissues of naïve (UIL), yeast-injected (YIL), and saline-injected (SIL) larvae of the blowfly Chrysomya megacephala, using the NADPH diaphorase technique. Our findings revealed similar levels of NOS activity in muscle, fat body, Malpighian tubule, gut, and brain, suggesting that NO synthesis may not be involved in the immune response of these larval systems. These results were compared to many studies that recorded the involvement of NO in various physiological functions of insects.

Anti-Saccharomyces cerevisiae antibody is not useful to differentiate between Crohn's disease and intestinal tuberculosis in India.

CONTEXT: Clinical, endoscopic, radiological and histological parameters of intestinal tuberculosis (IT) and Crohn's disease (CD) are so similar that differentiation between these two diseases, which require different treatment, is difficult. Anti- Saccharomyces cerevisiae antibody (ASCA), which is often present in the sera of patients with CD, may be potentially useful to differentiate CD from IT. AIM: To evaluate the role of enzyme-linked immunosorbent assay test for ASCA in serum in differentiating CD from intestinal tuberculosis. SETTINGS AND DESIGN: Prospective case-control study. MATERIALS AND METHODS: Sixteen patients with IT, 16 CD, 36 UC diagnosed using standard parameters and 12 controls (11 healthy subjects and one with colonic carcinoma) were tested for IgG ASCA in serum. STATISTICAL ANALYSIS USED: Categorical variables were analyzed using Chi-square test with Yates' correction, as applicable. Continuous variables were analyzed using Mann-Whitney U test. RESULTS: Eight of 16 (50%) patients with IT, 10 of 16 with CD (62%), nine of 35 with UC (26%) and one of 12 controls tested positive for ASCA in serum. Though the frequency of ASCA in serum was comparable among patients with IT and CD (8/16 vs. 10/16, P = ns), IT and UC (8/16 vs. 9/35, P =ns), CD and UC (10/16 vs. 9/35, P =ns), its frequency in CD or IT but not in UC was higher than healthy controls (P Conclusions: Serum ASCA is unlikely to be useful to differentiate between CD and IT in India.

Prevalencia de marcadores serológicos ANCA y ASCA en una población con colitis ulcerosa / Presence of antineutrophil cytoplasmic antibodies (ANCA) and anti Saccharomyces cerevisiae antibodies (ASCA) among patients with ulcerative colitis

Background: The diagnosis of inflammatory bowel disease is supported by clinical findings and complementary tests. The presence of specific serological markers could be helpful in the characterization of this condition. Aim: To assess the prevalence of ANCA and ASCA in a group of patients with ulcerative colitis (UC) and its association with clinical features. Material and Methods: Sixty four patients with UC in remission (age range 16-72 years, 33 males) were studied. In a venous blood sample ANCA were measured by indirect immunofluorescence and ASCA by enzyme immune assays for IgG and IgA. Results: Forty four percent of patients were positive for ANCA, 9 percent for ASCA and 6 percent for both markers. There was a significant correlation between the presence of ANCA and duration of the UC (<5 years 50 percent, 5-10 years 42.9 percent, 15 years 30 percent) and the number of crises (one crises 31 percent, 2-5 crises 51.9 percent and >5 crises 87.5). The proportion of colectomized patients with positive ANCA was higher (57.1 percent). Conclusions: The prevalence of ANCA in the studied population is similar to the published data. The presence of ANCA was significantly higher in UC patients with shorter evolution, higher number of crises and in those with a history of colectomy. There was a low prevalence of ASCA positive patients.

A predictive model for the level of sIgA based on IgG levels following the oral administration of antigens expressed in Sacchromyces cerevisiae

Oral vaccination may be the most efficient way of inducing an immune response at the remote mucosal site through the common mucosal immune network. Antigenspecific secretory IgA (sIgA) is the major immunoglobulin type generally detected in the secretions of experimental animals following an effective oral immunization. Actinobacillus pleuropneumoniae causing disease in the lung of pig initially interacts, colonizes, and infects the host tissues at the mucosal surface of the respiratory tract. Also, importantly for A. pleuropneumoniae protection, the quantity of sIgA in the lung had merits associated with the mucosal immunity. However, there is no simple method to monitor the level of sIgA as an indicator for the induction of local immune responses by an oral vaccination in the target tissue. Therefore, the relationship between sIgA and IgG was analyzed to evaluate the induction of local immune responses by an oral immunization with Saccharomyces cerevisiae expressing the apxIA and apxIIA genes of A. pleuropneumoniae in this study. The correlation coefficient of determination (r2 x 100) for paired samples in both vaccinated and control groups showed a significant positive-relationship between IgG in sera and sIgA in the lung or intestine. These results indicated that IgG antibody titers in sera could be useful to indirectly predict local immune response, and sIgA, in the lung or intestine to evaluate the efficacy of an oral vaccination.

Comparison of Behcet's Disease and Recurrent Aphthous Ulcer According to Characteristics of Gastrointestinal Symptoms

Behcet's disease (BD) is a multisystemic chronic inflammatory disease. It is characterized by recurrent oral and genital ulcers, uveitis, skin lesions and other manifestations, including neurologic, vascular, joint, and gastrointestinal ulcers of variable severity. Recurrent aphthous ulcer (RAU) represents a very common, but poorly understood, mucosal disorder. If a patient of RAU without any other typical symptoms of BD has gastrointestinal symptoms, it is difficult to distinguish this RAU from true BD with gastrointestinal involvement. Because pathognomonic clinical features and tools are absent, the differential diagnosis of these two diseases relies on the characteristic clinical features and the judgement of an experienced physician. Sixty-five out of a total 960 RAU patients and forty-four of 556 BD patients with gastrointestinal symptoms between January 1996 and December 2003 participated in this study. All were evaluated with esophagogastroduodenoscopy and colonoscopy. Clinical, endoscopic and histopathologic findings were analyzed and ELISA tests were conducted to detect serum levels of ASCA and pANCA. No significant difference was found between the two groups. Differential diagnosis between RAU with gastrointestinal symptoms and BD with gastrointestinal involvement requires further prospective, large-scale study.

Diagnostic Role of Anti-Saccharomyces cerevisiae and Antineutrophil Cytoplasmic Autoantibodies in Pediatric Inflammatory Bowel Disease / 대한소화기학회지

BACKGROUND/AIMS: Combined measurement of perinuclear antineutrophil cytoplasmic autoantibodies (pANCA) and anti-Saccharomyces cereviseae mannan antibodies (ASCA) has recently been suggested as a valuable diagnostic approach to inflammatory bowel disease (IBD) in the pediatric age group. The aim of this study was to test the accuracy of the assay using pANCA and ASCA in diagnosing pediatric ulcerative colitis (UC) and Crohn's disease (CD). METHODS: Sera were collected from 25 patients with IBD (17 with CD, 8 with UC) and 32 healthy controls. The levels of pANCA and ASCA were determined by using a standard indirect immunofluorescence technique on ethanol-fixed granulocytes and an ELISA assay, respectively. RESULTS: In patients with UC, the sensitivity, specificity, and positive predictive value of the pANCA test were 38%, 88%, and 60%, respectively. Such values were not changed significantly in the case of positive pANCA and negative ASCA. The sensitivity, specificity, and positive predictive value of ASCA test in diagnosing CD were 71%, 88%, and 92%, respectively. The combination of pANCA negative and ASCA positive was not significant. CONCLUSIONS: ASCA and pANCA assays are highly disease specific for CD and UC, respectively. These serological tests can assist clinicians in diagnosing and categorizing patients with IBD and may be useful in making therapeutic decisions.

Expressao do antigeno de 18kDa de Mycobacterium leorae na levedura Saccaromyces cerevisisae e estudo das propriedades imunogenicas do antigeno recombinante / Expression of the antigen of 18kDa of Mycobacterium leprae na levedura Saccharomyces cerevisiae and study of imonogenics properties of recombinant antigen

Dois diferentes sistemas de expressao do antigeno de 18 kDa de M. leprae (p18) em S. cerevisiae, um intracelular e outro de secrecao, foram desenvolvidos. Ambos os sistemas mostraram-se efetivos para a expressao, mas a purificacao da p18 secretada mostrou-se mais simples. Comparando diferentes cepas hospedeiras e condicoes de cultivo, foi obtido um sistema de secrecao de alto rendimento (mais de 100 mg de proteina biologicamente ativa por litro). A p18 foi purificada do meio de cultura da levedura por precipitacao, seguida de cromatografias de troca ionica e por filtracao em gel. As propriedades imunologicas da proteina recombinante, nativa ou previamente irradiada com raios 'GAMA' foram analisadas em camundongos. Ambas as preparacoes desencadearam producao de anticorpos e reacao de hipersensibilidade tardia, correspondentes as respostas humoral e celular, respectivamente. Em adicao, a irradiacao previa do antigeno potencializou sua imunogenicidade a nivel celular. Estes resultados demonstram ser esta proteina forte candidata para utilizacao em novos testes cutaneos para a monitorizacao da resposta celular contra M. leprae

Expression of Mycobacterium leprae 18-kDa antigen in yeast in a GPI-anchored form

The 18-kDa protein from Mycobacterium leprae is a major target for the immune response in leprosy. We have developed a system to express this antigen in yeast as a fusion protein with the C-terminal region of the yeast membrane protein GAS1, which would render the recombinant protein anchored to the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. Cells lacking the GAS1 gene and transformed with the hybrid 18-kDa-GAS1 construct express a polypeptide that reacts with an 18-kDa-specific monoclonal antibody. In addition, these cells react with an alpha-CRD antibody after GPI-PLC treatment. The non-transformed cells are negative. These data indicate that our system may be suitable for the expression of foreign proteins in yeast in a GPI-anchored form

Isolation of temperature-sensitive yeast GPI-anchoring mutants

We are using a genetic approach to explore the synthesis and function of glycosylphosphatidylinositol (GPI). We have developed a novel strategy to isolate Saccharomyces cerevisiae mutants blocked in GPI anchoring by screening colonies of mutagenized yeast cells for those that fail to incorporate [3H]inositol into protein. Among our isolates are strains blocked in mannosylation of the GPI-anchor precursor, and strains defective in the synthesis of N-acetylglucosaminylphosphatidylinositol (GlcNAc-PI). We have characterized one mutant, gpil, further. This strain is defective in GlcNAC-PI synthesis and is temperature-sensitive for growth. Completion of the first step in GPI assembly is therefore required for the growth of the unicellular eukaryote S. cerevisiae. We have isolated plasmids that complement the gpil mutation from S. cerevisiae genomic DNA- and fission yeast cDNA libraries

Estudo da atividade antimicrobiana de extrato de folha de Aristolochia gigantea Mart e Zucc / Study of the antimycrobial activity of extracts from the leaves of Aristolochia gigantea Mart and Zucc

Foram investigadas as atividades antibacteriana e antifúngica dos extratos éter de petróleo e alcaloídico de folhas de Aristolochia gigantea Mart e Zucc, pelo método de difusäo em agar. O extrato éter de petróleo mostrou atividade contra uma bactéria Gram-positiva. O extrato alcaloídico apresentou atividade contra duas bactérias Gram-positivas. Uma Gram-negativa, näo apresentando qualquer atividade antifúngica.