Interleukin-10 but not Transforming Growth Factor beta inhibits murine activated macrophages Paracoccidioides brasiliensis killing: Effect on H2O2 and NO production

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Role of TLR2 and TLR4 in Human Neutrophil Functions Against Paracoccidioides brasiliensis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Interactions in the Agrobacterium-soybean system and capability of some Brazilian soybean cultivars to produce somatic embryos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

SEROLOGICAL RESPONSE OF CHICKENS TO INFECTION WITH SALMONELLA-GALLINARUM S-PULLORUM DETECTED BY ENZYME-LINKED-IMMUNOSORBENT-ASSAY

The serological response to Salmonella pullorum and S. gallinarum infection in chickens was studied with an indirect enzyme-linked immunosorbent assay (ELISA). In broiler chickens, a more virulent strain of S. pullorum produced a significantly lower serum IgG titer than did a less virulent strain. In laying hens, the serum and egg-yolk IgG titers were very similar. In chickens infected with S. gallinarum, high IgG titers persisted for 30 weeks. In chickens reinfected with this strain, each reinfection was followed by transitory increases in IgG lasting no longer than 2 weeks. Serum samples from Brazil taken from a laying flock with evidence of fowl typhoid showed much higher antibody levels than did those from three uninfected flocks. Using lipopolysaccharide as the detecting antigen, infections caused by these salmonellae could be differentiated from those caused by other groups. Incorporation of the appropriate flagella antigen in the ELISA allowed differentiation between infections caused by S. pullorum and S. enteritidis.

Role of Paracoccidioides brasiliensis cell wall fraction containing beta-glucan in tumor necrosis factor-alpha production by human monocytes: correlation with fungicidal activity

The polysaccharide fraction of Paracoccidioides brasiliensis mycelial cell wall (F1 fraction), the active component of which is composed of beta-glucan, was investigated in regard to the activation of human monocytes for fungal killing. The cells were primed with interferon-gamma (IFN-gamma) or F1 (100 and 200 mug ml(-1)) or F1 (100 and 200 mug ml(-1)) plus IFN-gamma for 24 h and then evaluated for H2O2 release. In other experiments, the cells were pretreated with the same stimuli, challenged with a virulent strain of P. brasiliensis and evaluated for fungicidal activity and levels of tumor necrosis factor (TNF-alpha) in the supernatants. F1 increased the levels of H2O2 in a similar manner to IFN-gamma. However, a synergistic effect between these two activators was not detected. on the contrary, a significant fungicidal activity was only obtained after priming with IFN-gamma plus F1. This higher activity was associated with high levels of TNF-alpha in the supernatants of the cocultures. Overall, P. brasiliensis F1 fraction induced human monocytes to release relatively high levels of TNF-alpha, which, in combination with IFN-gamma, is responsible for the activation of human monocytes for effective killing of P. brasiliensis.

Antioxidant activity of the melanin pigment extracted from Aspergillus nidulans

Melanins are pigments of high molecular weight formed by oxidative polymerization of phenolic or indolic compounds. A number of fungi, including Aspergillus nidulans, produce pigments related or identical to melanin, which are located on cell walls or exist as extracellular polymers. The aim of the present study was to assess the antioxidant activity of synthetic melanin and of the pigment extracted from the mycelium and culture medium after growth of the highly melanized strain (MEL1) from A. nidulans. The ability of the melanin pigment to scavenge the oxidants HOCl and H2O2 was evaluated by inhibition of the oxidation of 5-thio-2-nitrobenzoic acid (TNB) using several melanin concentrations. The results showed that the pigment of the MEL1 strain competes with TNB for H2O2 and HOCl, inhibiting TNB oxidation in a concentration-dependent manner. For the HOCl oxidant, this inhibition was comparable to that of synthetic melanin, whose IC50 values were quite close for both pigments. Thus, our results suggest that the melanin from A. nidulans is a potential HOCl scavenger and may be considered a promising material for the cosmetic industry for the formulation of creams that protect the skin against possible oxidative damage.

Effect of prostaglandins on the production of H2O2 and cytokines that modulate the fungicidal activity of human monocytes against Paracoccidioides brasiliensis

Human monocytes lack fungicidal activity against high virulent strains of Paracoccidioides brasiliensis, the etiological agent of paracoccidioidomycosis, even after IFN-γ activation. However, monocytes treated with indomethacin (INDO) or INDO plus IFN-γ effectively killed this fungus, suggesting an inhibitory role of prostaglandins in this process. Thus, the purpose of this work was to test if this regulatory effect of prostaglandin was associated with alterations on H2O2 production and/or on modulatory cytokines levels, such as TNF-α, IL-10, and IL-6. Peripheral blood monocytes obtained from 10 healthy donors were incubated for 18 hours in the presence or absence of IFN-γ, INDO, or IFN-γ plus INDO, and further challenged with a high virulent strain of P. brasiliensis (Pb18) for 4 hours. Then, the monocytes cultures were evaluated for H2O2 release and fungicidal activity calculated by counting the colony forming units after plating. Moreover, on supernatants of the same cultures, TNF-α, IL-10, IL-6, and PGE2 concentrations were evaluated by ELISA. Monocytes treated with INDO or INDO plus IFN-γ presented higher fungicidal activity associated with the release of higher levels of H2O2 and TNF-α, but lesser levels of PGE2, when compared to nontreated cells. However, the levels of IL-10 and IL-6 were similar between treated and nontreated cells. The results suggest that human monocytes when challenged with high virulent strains of P. brasiliensis produce prostaglandins that inhibit the fungicidal activity of these cells by reducing H2O2 and TNF-α levels.

Invasin of Yersinia pseudotuberculosis is not a polyclonal activator of mouse B lymphocytes

It is known that the invasin molecule of Yersinia pseudotuberculosis stimulates human peripheral B cells in vitro. In this work we evaluated the in vivo role of invasin as polyclonal activator of B lymphocytes in the mouse experimental model, by comparing strains of Y. pseudotuberculosis expressing invasin and isogenic inv mutants. Swiss mice were infected intravenously with two strains expressing invasin (YpIII pIB1 and an isogenic virulence plasmid-cured strain, YpIII) and with two invasin mutant strains (Yp100 pIB1 and Yp100, plasmid-cured). Spleen cells were sampled on days 7, 14, 21 and 28 after infection. Immunoglobulin (Ig)-secreting spleen cells were detected by protein A plaque assay and specific antibodies were detected in sera by ELISA. The virulent strain YPIII pIB1 (wild type) did not provoke polyclonal activation of B lymphocytes in vivo. In general, fewer Ig-secreting spleen cells of all isotypes were found in the infected animals than in the control animals. Specific IgG antibodies were detected in the sera of animals infected with all strains. The peak response occurred on the 21 st day post-infection, and the Yp100 strain provoked the highest level of these antibodies. We concluded that invasin is not a polyclonal activator of murine B cells.

Use of reverse transcriptase polymerase chain reaction (RT-PCR) in molecular screening of Newcastle disease virus in poultry and free-living bird populations

The aim of this study was to evaluate a simple molecular method of reverse transcriptase polymerase chain reaction (RT-PCR) to differentiate Newcastle disease virus strains according to their pathogenicity, in order to use it in molecular screening of Newcastle disease virus in poultry and free-living bird populations. Specific primers were developed to differentiate LaSota-LS-(vaccine strain) and Sao Joao do Meriti-SJM-strain (highly pathogenic strain). Chickens and pigeons were experimentally vaccinated/infected for an in vivo study to determine virus shedding in feces. Validation of sensitivity and specificity of the primers (SJM and LS) by experimental models used in the present study and results obtained in the molecular analysis of the primers by BLAST made it possible to generalize results. The development of primers that differentiate the level of pathogenicity of NDV stains is very important, mainly in countries where real-time RT-PCR is still not used as a routine test. These primers were able to determine the presence of the agent and to differentiate it according to its pathogenicity. © 2012 Springer Science+Business Media B.V.

Efeito de citocinas sobre a atividade fungicida e fungistática de neutrófilos humanos contra o Paracoccidioides brasiliensis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Infecção experimental de gatas gestantes com cistos teciduais de toxoplasma gondii de diferentes genótipos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito de Interleucina-6 sobre a atividade fungicida e produção de citocinas por monócitos humanos, infectados in vitro com cepa virulenta de paracoccidioides brasiliensis

Pós-graduação em Patologia - FMB

Papel dos receptores Toll-Like na atividade dos neutrófilos humanos desafiados com a cepa de alta virulência do Paracoccidioides brasiliensis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeito de prostaglandinas sobre a atividade fingicida de monócitas humanos desafiados com o Paracoccidioides brasiliensis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Evaluation of the presence of microorganisms in solid-organ preservation solution

OBJECTIVE: To assess the presence of microorganism contamination in the preservation solution for transplant organs (kidney/pancreas). Method: Between August 2007 and March 2008, 136 samples of preservation solution were studied prior to graft implantation. Variables related to the donor and to the presence of microorganisms in the preservation solution of organs were evaluated, after which the contamination was evaluated in relation to the recipient culture variable. Univariate and multivariate statistical analyses were performed. RESULTS: The contamination rate of the preservation solution was 27.9%. Coagulase-negative Staphylococcus was the most frequently isolated microorganism. However, highly virulent agents, such as fungi and enterobacteria, were also isolated. In univariate analysis, the variable donor antibiotic use was significantly associated to the contamination of the preservation solution. on the other hand, multivariate analysis found statistical significance in donor antibiotic use and donor's infectious complications variables. CONCLUSIONS: In this study, 27.9% of the preservation solutions of transplant organs were contaminated. Infectious diseases and non-use of antibiotics by the donor were significantly related to the presence of microorganisms in organ preservation solutions. Contamination in organ preservation solutions was not associated with infection in the recipient.