Evaluation of gene expression levels for cytokines in ocular toxoplasmosis.

This study evaluated levels for mRNA expression of 7 cytokines in ocular toxoplasmosis. Peripheral blood mononuclear cells (PBMC) of patients with ocular toxoplasmosis (OT Group, n = 23) and chronic toxoplasmosis individuals (CHR Group, n = 9) were isolated and stimulated in vitro with T. gondii antigen. Negative controls (NC) were constituted of 7 PBMC samples from individuals seronegative for toxoplasmosis. mRNA expression for cytokines was determined by qPCR. Results showed a significant increase in mRNA levels from antigen stimulated PBMCs derived from OT Group for expressing IL-6 (at P < .005 and P < .0005 for CHR and NC groups, respectively), IL-10 (at P < .0005 and P < .005 for CHR and NC groups, respectively) and TGF-ß (at P < .005) for NC group. mRNA levels for TNF-α and IL-12 were also upregulated in patients with OT compared to CHR and NC individuals, although without statistical significance. Additionally, mRNA levels for IL-27 and IFN-γ in PBMC of patients with OT were upregulated in comparison with NC individuals. Differences between OT and NC groups were statistically significant at P < .05 and P < .0005, respectively.

Interleukin-18, interleukin-12B and interferon-γ gene polymorphisms in Brazilian patients with rheumatoid arthritis: a pilot study.

Polymorphisms in interleukin (IL)-18, IL-12 and interferon (IFN)-γ genes are associated with different levels of cytokines expression and have been associated with rheumatoid arthritis (RA). IL-18 +105 A/C, IL-12B +1188 A/C and IFN-γ +874 T/A polymorphisms were analyzed by restriction fragment length polymorphism-polymerase chain reaction (PCR) and amplification refractory mutation system PCR from 90 RA patients and 186 healthy individuals. There were significant differences to IL-18 +105 A/C polymorphism between the RA and control groups (odds ratio = 3.77; P < 0.0001). Individual carriers of the variant allele C had a 3.77-fold increased risk of for RA (P = 0.0032). No association was observed for IL-12B and IFN-γ polymorphisms. Our finds suggest a possible role for IL-18 polymorphism in the RA susceptibility in studied population.

Pulmonary cryptococcosis in childhood systemic lupus erythematosus and Sjögren syndrome overlap: a rare opportunistic infection.

Meningitis is the main manifestation of cryptococcosis in adult systemic lupus erythematosus (SLE) patients, and other organs and systems, such as the lungs, are rarely affected in this fungal infection. To our knowledge, no case of pulmonary cryptococcosis has been described in the pediatric lupus population. Therefore, we report herein one patient with childhood SLE (C-SLE) and Sjögren's syndrome overlap that presented encapsulated Cryptococcus yeast cells in lung tissue. A 14-year-old girl was diagnosed with C-SLE. At the age of 16 years and 5 months, she presented with fever, cough and dyspnea, without headache, vomiting, and also without signs of meningeal irritation or other clinical manifestations. She was being treated with mycophenolate mofetil, hydroxychloroquine and prednisone. Chest radiography and chest computer tomography showed a single nodule in the left posterior apex and three nodular lesions in the left hemithorax respectively. Bronchoalveolar lavage and transbronchial biopsy were normal and without isolation of bacteria or fungi. Voriconazole was empirically introduced for 21 days. Fifteen days after the first biopsy, she underwent open thoracotomy with surgical left lung biopsy and was diagnosed with pulmonary cryptococcosis. Voriconazole was replaced with oral fluconazole and this antifungal therapy was maintained with improvement of clinical manifestations and without marked alteration of radiological images. In conclusion, we report the first case of pulmonary cryptococcosis in Sjögren's and C-SLE patient with a satisfactory clinical response to antifungal therapy. Fungal infections should be excluded in the presence of lung nodules and etiological identification is required for proper treatment.

Intestinal parasitic infection and associated risk factors, among children presenting at outpatient clinics in Manaus, Amazonas state, Brazil.

In a cross-sectional study, the prevalences of intestinal parasitic infection and the associated risk factors were explored among children aged 0-10 years attending outpatient clinics in Manaus, the capital city of the north Brazilian state of Amazonas. Data indicating socio-economic level and demographic factors were collected in interviews with the accompanying parent or guardian of each child. Parasitic infections were detected by the microscopical examination of faecal samples, while nutritional status was categorized by reference to the growth curves published by the U.S. National Center for Health Statistics. The data collected were analysed in a mixed, logistic-regression model. Of the 451 children tested, 58.7% were found to have intestinal parasitic infection, the most prevalent parasites being Giardia lamblia (21.5%), Endolimax nana (17.9%), Entamoeba histolytica/Entamoeba dispar (13.7%), Ascaris lumbricoides (13.5%) and Trichuris trichiura (4.9%). The prevalences of stunting, underweight and wasting among the children were 17.5%, 14.7% and 9.8%, respectively. There was no association between intestinal parasite infection and any of these indicators of malnutrition. Children whose accompanying parent/guardian was poorly educated and the elder children investigated were found to be at relatively high risk of intestinal parasitic infection (P<0.05 for each). Intestinal parasitic infections still clearly represent an important public-health problem in the northern region of Brazil.

Effect of culture conditions on lipase production by Fusarium solani in batch fermentation.

Lipase (Glycerol ester hydrolase EC 3.1.1.3.) from a Brazilian strain of Fusarium solani FSI has been investigated. The effect of different carbon sources and trace elements added to basal medium was observed with the aim of improving enzyme production. Lipase specific activity was highest (0.45 U mg(-1)) for sesame oil. When this medium was supplemented with trace elements using olive oil, corn oil and sesame oil the lipase specific activity increased to 0.86, 1.89 and 1.64 U mg(-1), respectively, after 96 h cultivation without any considerable biomass increase. The Km of this lipase using pNPP (p-nitrophenylpalmitate) as substrate, was 1.8 mM with a Vmax of 1.7 micromol min(-1) mg protein(-1). Lipase activity increased in the presence of increasing concentrations of hexane and toluene. In contrast, incubation of this enzyme with water-soluble solvents decreased its activity after 10% concentration (v/v) of the solvent. The lipase activity was stable below 35 degrees C but above this temperature activity losses were observed.

[Anti-HBs antibodies from rabbits and their use as reagents for serological tests in the diagnosis of hepatitis B]. / Anticorpos anti-HBs obtidos em coelhos e sua utilizacão como reagentes para testes sorológicos no diagnóstico da hepatite B.

By the method of affinity chromatography a partially purified antigen was obtained after passing the plasma of an asymptomatic carrier of HBsAg through a column of Sepharose 4B linked to angi-HBs. This antigen was inoculated in rabbits using a schedule of 1,0 mg in the first dose and 4 other doses of 0,5 mg with intervals of approximately 15 days. Observing that blood samples collected after the 5th inoculation showed no change in antibody levels, the animals were bled on the 62th day and these immune sera were standardized with the following tests for the detection of HBsAg: Reverse passive hemagglutination (R-PHA) - using specific gamma globulin that was obtained from rabbit sera by affinity chromatography and reaching an optimal concentration of 10 micrograms/ml to sensitise SRBC at 5% fixed in glutaraldehyde. Counter immuno electrophoresis (CIEP) - using the rabbit immune sera diluted to 1/20 as a reagent for the detection of HBsAg. The immune sera was also used to conjugate new Sepharose 4B for affinity chromatography and was found having a linking capacity of approximately 0,5 to 1,0 mg of HBsAg per ml of Sepharose after complete saturation.

Hepatic cholesteryl ester metabolism in reptiles. A comparative study of three species of Brazilian lizards.

Cholesterol esterase (CEase) and acylcoenzyme A: cholesterol acyltransferase (ACATase) activities were identified in liver cytoplasmatic extracts from Tropidurus torquatos (Iguanidae), Ameiva ameiva (Teiidae) and Hemidactylus mabouia (Gekkonidae). Optimum conditions were established to measure the hydrolytic activity of CEase and esterifying activities of CEase and ACATase. The activities of both enzymes were generally similar in all three species of reptiles, and did not differ greatly from values reported for a variety of mammalian species.

Anticorpos anti-HBs obtidos em coelhos e sua utilizacao como reagentes para testes sorologicos no diagnostico da hepatite B. / Anti-HBs antibodies obtained in rabbits and its use as reagents for serological tests in the diagnosis of hepatitis B

Pela tecnica de cromatografia de afinidade utilizando-se a resina de Sepharose 4B ligada ao anti-HBs, obteve-se na passagem de plasma de portador assintomatico de antigeno HBs, um antigeno parcialmente purificado. Este antigeno foi utilizado para a inoculacao em coelhos, num esquema de cinco doses, sendo a primeira dose de 1 mg e as quatro subsequentes de 0,5 mg, com intervalos aproximadamente de quinze dias.Obsevando-se que os titulos nao mais variaram apos a quinta inoculacao os animais foram sangrados no 62o dia e os anticorpos anti-HBs obtidos foram padronizados atraves dos seguintes metodos para deteccao de antigeno HBs: a) Hemaglutinacao passiva reversa (HAPR) - utilizando-se a gamaglobulina especifica obtida de soro imune dos coelhos atraves de cromatografia de afinidade, alcancando uma concentracao otima de apenas 10 micros/ml para a sensibilizacao de hemacias de carneiro a 5%, fixadas com glutaraldeido. b) Contraimunoeletroforese (CIEF) - utilizando-se o soro imune diluido ate 1/20 como reagente para a deteccao do antigeno HBs. O soro imune anti-HBs foi tambem utilizado para a conjuncao com uma nova resina de Sepharose 4B tendo uma captacao aproximada de 0,5 a 1,0 mg de antigeno HBs por ml de resina apos completa saturacao