BINDING OF PARACOCCIDIOIDES-BRASILIENSIS TO LAMININ THROUGH SURFACE GLYCOPROTEIN GP43 LEADS TO ENHANCEMENT OF FUNGAL PATHOGENESIS

Extracellular matrix protein laminin binds specifically to yeast forms of Paracoccidioides brasiliensis and enhances adhesion of the fungus to the surface of epithelial Madin-Darby canine kidney cells in vitro. Immunoblotting of fungal extracts showed that the gp43 glycoprotein is responsible for adhesion. This was confirmed by binding assays using purified gp43, with a K-d of 3.7 nM. The coating of P. brasiliensis yeast forms with laminin before injection into hamster testicles enhanced the fungus virulence, resulting in a faster and more severe granulomatous disease. These results indicate that interaction of fungi with extracellular matrix elements may constitute a basis for the evolution of fungal infection toward regional spreading and dissemination.

Circadian rhythm of anti-fungal prenylated chromene in leaves of Piper aduncum

Leaves of Piper aduncum accumulate the anti-fungal chromenes methyl 2,2-dimethyl-2H-1-chromene-6-carboxylate (1) and methyl 2,2-dimethyl-8-(3'-methyl-2'-butenyl)-2H-1-chromene-6-carboxylate (2). The enzymatic formation of 2 from dimethylallyl diphosphate and I was investigated using cell-free extracts of the title plant. An HPLC assay for the prenylation reaction was developed and the enzyme activity measured in the protein extracts. The prenyltransferase that catalyses the transfer of the dimethylallyl group to C-2' of 1 was soluble and required dimethylallyl diphosphate as the prenyl donor. In the leaves, the biosynthesis of the prenylated chromene 2 was time-regulated and prenyltransferase activity depended upon circadian variation. Preliminary characterisation and purification experiments on the prenyltransferase from P. aduncum have been performed. Copyright (C) 2005 John Wiley & Sons, Ltd.

Influence of agricultural biomass burning on aerosol size distribution and dry deposition in southeastern Brazil

The size distributed composition of ambient aerosols is used to explore seasonal differences in particle chemistry and to show that dry deposition fluxes of soluble species, including important plant nutrients, increase during periods of biomass (sugar cane trash) burning in São Paulo State, Brazil. Measurements were made at a single site centrally located in the State's sugar cane growing region but away from the immediate vicinity of burns, so that the air sampled was representative of the regional background. Calculation of ion equivalent balances showed that during burning periods smaller particles (Aitken and accumulation modes) were more acidic, containing higher concentrations of SO(4)(2-), oxalate, NO(3)(-), HCOO(-), CH(3)COO(-), and Cl(-), but insufficient NH(4)(+) and K(+) to achieve neutrality. Larger particles showed an anion deficit due to the presence of unmeasured ions and comprised resuspended dusts modified by accumulation of nitrate, chloride, and organic anions. Increases of resuspended particles during the burning season were attributed to release of earlier deposits from the surfaces of burning vegetation as well as increased vehicle movement on unsurfaced roads. During winter months the relative contribution of combined emissions from road transport and industry diminished due to increased emissions from biomass combustion and other activities specifically associated with the harvest period. Positive increments in annual particulate dry deposition fluxes due to higher fluxes during the sugar cane harvest were 44.3% (NH(4)(+)), 42.1 % (K(+)), 31.8% (Mg(2+)), 30.4% (HCOO(-)), 12.8% (Cl(-)), 6.6% (CH(3)COO(-)), 5.2% (Ca(2+)), 3.8% (SO(4)(2-)), and 2.3% (NO(3)(-)). Na(+) and oxalate fluxes were seasonally invariant. Annual aerosol dry deposition fluxes (kg ha(-1)) were 0.5 (Na(+)), 0.25 (NH(4)(+)), 0.39 (K(+)), 0.51 (Mg(2+)), 3.19 (Ca(2+)), 1.34 (Cl(-)), 4.47 (NO(3)(-)), 3.59 (SO(4)(2-)), 0.58 (oxalate), 0.71 (HCOO(-)), and 1.38 (CH(3)COO(-)). Contributions of this mechanism to combined aerosol dry deposition and precipitation scavenging (inorganic species, excluding gaseous dry deposition) were 31% (Na(+)), 8% (NH(4)(+)), 26% (K(+)), 63% (Mg(2+)), 66% (Ca(2+)), 32% (Cl(-)), 33% (NO(3)(-)), and 36% (SO(4)(2-)).

Energy balance optimization of sugarcane crop residual biomass

This work presents a mathematical model for helping mills choose sugarcane varieties for planting. It maximizes crop residual biomass energy balance by considering the difference between generated and consumed energy in the process of transferring this biomass from the field to the processing center; it takes into account enterprise demand restrictions and cane planting area. For this full zero-one linear programming techniques were proposed. The model is viable for choosing sugarcane varieties that would benefit sugarcane production and industrial systems, by reducing crop residue and increasing final energy production. (c) 2006 Published by Elsevier Ltd.

Glyceraldehyde-3-phosphate dehydrogenase of Paracoccidioides brasiliensis is a cell surface protein involved in fungal adhesion to extracellular matrix proteins and interaction with cells

The pathogenic fungus Paracoccidioides brasiliensis causes paracoccidioidomycosis, a pulmonary mycosis acquired by inhalation of fungal airborne propagules, which may disseminate to several organs and tissues, leading to a severe form of the disease. Adhesion to and invasion of host cells are essential steps involved in the infection and dissemination of pathogens. Furthermore, pathogens use their surface molecules to bind to host extracellular matrix components to establish infection. Here, we report the characterization of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of P. brasiliensis as an adhesin, which can be related to fungus adhesion and invasion. The P. brasiliensis GAPDH was overexpressed in Escherichia coli, and polyclonal antibody against this protein was obtained. By immunoelectron microscopy and Western blot analysis, GAPDH was detected in the cytoplasm and the cell wall of the yeast phase of P. brasiliensis. The recombinant GAPDH was found to bind to fibronectin, laminin, and type I collagen in ligand far-Western blot assays. of special note, the treatment of P. brasiliensis yeast cells with anti-GAPDH polyclonal antibody and the incubation of pneumocytes with the recombinant protein promoted inhibition of adherence and internalization of P. brasiliensis to those in vitro-cultured cells. These observations indicate that the cell wall-associated form of the GAPDH in P. brasiliensis could be involved in mediating binding of fungal cells to fibronectin, type I collagen, and laminin, thus contributing to the adhesion of the microorganism to host tissues and to the dissemination of infection.

Influence of water level variation on biomass and chemical composition of the aquatic macrophyte Eichhornia azurea (Kunth) in an oxbow lake of the Rio Mogi-Guacu (São Paulo, Brazil)

The influence of water level variation (flood pulse) on the biomass and chemical composition of the aquatic macrophyte Eichhornia azurea, was investigated in a tropical oxbow lake of the Rio Mogi-Guacu, State of São Paulo, Brazil. The flood pulse causes an increase in total nitrogen content from 0.67 to 1.35 mg/L and total phosphorus content from 10.5 to 101.0 mu g/L of the water. This fertilization, associated with other factors, determines a typical seasonal variation in the biomass and chemical composition of the macrophyte.

Biomass production and secretion of hydrolytic enzymes are influenced by the structural complexity of the nitrogen source in Fusarium oxysporum and Aspergillus nidulans

The structural complexity of the nitrogen sources strongly affects biomass production and secretion of hydrolytic enzymes in filamentous fungi. Fusarium oxysporum and Aspergillus nidulans were grown in media containing glucose or starch, and supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids), peptides (peptone) and protein (gelatin). In glucose, when the initial pH was adjusted to 5.0, for both microorganisms, higher biomass production occurred upon supplementation with a nitrogen source in the peptide form (peptone and gelatin). With a close to neutrality pH, biomass accumulation was lower only in the presence of the ammonium salt. When grown in starch, biomass accumulation and secretion of hydrolytic enzymes (amylolytic and proteolytic) by Fusarium also depended on the nature of the nitrogen supplement and the pH. When the initial pH was adjusted to 5.0, higher growth and higher amylolytic activities were detected in the media supplemented with peptone, gelatin and casamino acids. However, at pH 7.0, higher biomass accumulation and higher amylolytic activities were observed upon supplementation with peptone or gelatin. Ammonium sulfate and casamino acids induced a lower production of biomass, and a different level of amylolytic enzyme secretion: high in ammonium sulfate and low in casamino acids. Secretion of proteolytic activity was always higher in the media supplemented with peptone and gelatin. Aspergillus, when grown in starch, was not as dependent as Fusarium on the nature of nitrogen source or the pH. The results described in this work indicate that the metabolism of fungi is regulated not only by pH, but also by the level of structural complexity of the nitrogen source in correlation to the carbon source.

Rhodocyclus gelatinosus biomass for egg yolk pigmentation

Biomass of the photosynthetic bacterium Rhodocyclus gelatinosus was used at different levels in laying hens' rations as a xanthophyll source. Sixty-four hens were used in the experiment that investigated the effects of different biomass concentrations on weight gain, egg production, egg weight, and yolk color as compared with a control group that received no biomass supplementation in the ration. Yolk color was scored by means of a color fan. All concentrations tested were able to provide yolk color scores higher than those provided by the control group. The pigment deposition began after 24 h of administration and reached a plateau around the twentieth day. Each increase in the supplementation level led to an additional increase on yolk color scores. Yolk colors of all treatments that received R. gelatinosus biomass differed significantly from the control group and from each other, corroborating that the increase in the biomass supplementation had a positive effect on color increase. Body weight loss occurred in all treatments. Egg production did not increase with the biomass addition, while a significant increase in egg weight was observed in the treatments that received the product.

Evaluation of Rhodocyclus gelatinosus biomass for broiler pigmentation

We evaluated the influence of dietry inclusion of corn gluten meal, apocartenoic acid ethyl ester (APO-EE), canthaxanthin, and Rhodocylus gelatinosus R-1 biomass on broiler carcass color. These oxycarotenoid sources were used as pigment supplements to a basal ration containing yellow corn as the sole source of xnathophylls. Objective color values of L (lightness),C (chroma), and h (hue) were measured on skin and meat surfaces of broiler carcasses. on both surfaces, R. gelatinosus R-1 biomass oxycarotenoids enhanced the chroma values (color saturation), as compared to yellow corn xanthophylls, and tended to provide yellowness to broiler carcasses, whereas the APO-EE and canthaxanthin tended to provide redness. At the concentrations studied, R. gelatinosus R-1 biomass oxycarotenoids were less effective than APO-EE and canthaxanthin in enhancing color saturation. Lightness, chroma, and blue values did not differ significantly between males and females. However, skin showed significantly higher color saturation than meat in breast and thigh portions of the carcass.

Estimating the aboveground biomass and wood volume of savanna woodlands in Brazil's Pantanal wetlands based on allometric correlations

The quantity and distribution of vegetal biomass are important aspects to consider in ecosystem studies. However, little information is available about Brazil's Pantanal woodland savannas. This work involved the development of regression equations of the aerial biomass and wood volume of native tree species in a region of woodland savanna on Rio Negro farm in the Pantanal of Nhecolandia, Brazil. Samples were taken from 10 trees of each of five species: Protium heptaphyllum (Aub1.) Marchand, Magonia pubescens A. St.-Hil., Diptychandra aurantiaca Tul., Terminalia argentea Mart. and Zucc. and Licania minutiflora (Sagot) Fritsch and from a miscellaneous group of I I different species. Linear and nonlinear regression analyses were developed relating the diameter at breast height to the dry weight of wood, branches and leaves, wood volume and total aerial biomass. All the regressions showed a significance of P < 0.05 and an R-2 close to or above 0.8. The biomass curve predicted by linear regression analysis of the studied species was similar to the nonlinear regression, with the exception of L. minutiflora and the miscellaneous group. The breast height diameter proved a good choice for estimating biomass and wood volume. The estimated wood volume and biomass of the Pantanal woodland savanna is crucial information for understanding the carbon cycle and for ensuring the region's conservation and sustainable use. (c) 2006 Elsevier B.V. All rights reserved.

DECOMPOSITION RATES OF SPENT FUNGAL REFUSE OF ATTA-SEXDENS (HYMENOPTERA, FORMICIDAE) AND IMPLICATIONS FOR ESTIMATING FORAGE INTAKE BY LEAF-CUTTING ANTS

Using simulated ceramic refuse chambers, field decomposition studies were performed on the spent fungal refuse of the lead-cutting ant Atta sexdens rubropilosa. Refuse half life was estimated at 40 days, with complete decomposition at 100 days. These results suggest that the conversion-factor method used to estimate forage input into leaf-cutting ant colonies must be corrected for decomposition, or serious estimation errors will occur.

Monoclonal antibodies against the 43,000 da glycoprotein from Paracoccidioides brasiliensis modulate laminin-mediated fungal adhesion to epithelial cells and pathogenesis

The surface glycoprotein gp43, a highly immunogenic component of Paracoccidioides brasiliensis, is used in the serodiagnosis of paracoccidioidomycosis (PCM) and has recently been shown to specifically bind the extracellular matrix protein laminin, Binding to laminin induces the increased adhesion of the fungus to epithelial cells; a hamster testicle infection model has shown that the gp43-dependent binding of fungal cells to laminin enhances their pathogenicity in vivo. We report on the production and characterization of 12 monoclonal antibodies against the gp43 that recognize peptide sequences in the molecule detecting at least three different epitopes as well as different isoforms of this antigen. MAbs interfered in the fungal pathogenicity in vivo either by inhibiting or enhancing granuloma formation and tissue destruction, Results suggest that P. brasiliensis propagules may start infection in man by strongly adhering to human lung cells, Thus, laminin-mediated fungal adhesion to human lung carcinoma (A549) cells was much more intense than to Madin-Darby canine kidney cells (MDCK), indicating differences in binding affinity, Subsequent growth of fungi bound to the lung cells could induce the granulomatous inflammatory reaction characteristic of PCM. Both steps are greatly stimulated by laminin binding in infective cells expressing gp43.

Differential gene expression by Paracoccidioides brasiliensis in host interaction conditions: Representational difference analysis identifies candidate genes associated with fungal pathogenesis

Paracoccidioides brasiliensis causes infection by the host inhalation of airborne propagules of the mycelia phase of the fungus. These particles reach the lungs, and disseminate to virtually all organs. Here we describe the identification of differentially expressed genes in studies of host-fungus interaction. We analyzed two cDNA populations of P. brasiliensis, one obtained from infected animals and the other an admixture of fungus and human blood thus mimicking the hematologic events of the fungal dissemination. Our analysis identified transcripts differentially expressed. Genes related to iron acquisition, melanin synthesis and cell defense were specially upregulated in the mouse model of infection. The upregulated transcripts of yeast cells during incubation with human blood were those predominantly related to cell wall remodeling/synthesis. The expression pattern of genes was independently confirmed in host conditions, revealing their potential role in the infection process. This work can facilitate functional studies of novel regulated genes that may be important for the survival and growth strategies of P. brasiliensis in humans. (c) 2006 Elsevier Masson SAS. All rights reserved.

Cultivation of Rubrivivax gelatinosus in fish industry effluent for depollution and biomass production

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