Genomic DNA microarray comparison of gene expression patterns in Paracoccidioides brasiliensis mycelia and yeasts in vitro

Paracoccidioides brasiliensis is a thermally dimorphic fungus, and causes the most prevalent systemic mycosis in Latin America. Infection is initiated by inhalation of conidia or mycelial fragments by the host, followed by further differentiation into the yeast form. Information regarding gene expression by either form has rarely been addressed with respect to multiple time points of growth in culture. Here, we report on the construction of a genomic DNA microarray, covering approximately 25% of the genome of the organism, and its utilization in identifying genes and gene expression patterns during growth in vitro. Cloned, amplified inserts from randomly sheared genomic DNA (gDNA) and known control genes were printed onto glass slides to generate a microarray of over 12 000 elements. To examine gene expression, mRNA was extracted and amplified from mycelial or yeast cultures grown in semi-defined medium for 5, 8 and 14 days. Principal components analysis and hierarchical clustering indicated that yeast gene expression profiles differed greatly from those of mycelia, especially at earlier time points, and that mycelial gene expression changed less than gene expression in yeasts over time. Genes upregulated in yeasts were found to encode proteins shown to be involved in methionine/cysteine metabolism, respiratory and metabolic processes (of sugars, amino acids, proteins and lipids), transporters (small peptides, sugars, ions and toxins), regulatory proteins and transcription factors. Mycelial genes involved in processes such as cell division, protein catabolism, nucleotide biosynthesis and toxin and sugar transport showed differential expression. Sequenced clones were compared with Histoplasma capsulatum and Coccidioides posadasii genome sequences to assess potentially common pathways across species, such as sulfur and lipid metabolism, amino acid transporters, transcription factors and genes possibly related to virulence. We also analysed gene expression with time in culture and found that while transposable elements and components of respiratory pathways tended to increase in expression with time, genes encoding ribosomal structural proteins and protein catabolism tended to sharply decrease in expression over time, particularly in yeast. These findings expand our knowledge of the different morphological forms of P. brasiliensis during growth in culture.

Dissolved organic carbon in rainwater from areas heavily impacted by sugar cane burning

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

Influence of sugar cane burning on aerosol soluble ion composition in Southeastern Brazil

Seasonal variability in the major soluble ion composition of atmospheric particulate matter in the principal sugar cane growing region of central São Paulo State indicates that pre-harvest burning of sugar cane plants is an important influence on the regional scale aerosol chemistry. Samples of particulate matter were collected between April 1999 and February 2001 in coarse (> 3.5 mum) and fine (< 3.5 mum) fractions, and analysed for HCOO-, CH3COO-, C2C42-, SO42- . Results indicated that the principal sources of the aerosols investigated NO3-, Cl-, Na+, K+, NH4+, Mg2+ and Ca2+ were local or regional in nature (scale of tens to a few hundreds of km), and that differences between air masses of varying origins were small. Fine particles were typically acidic, containing secondary nitrates, sulphates and organic species. Coarse fraction concentrations were mainly influenced by physical parameters (wind speed, movement of vehicles and surface condition) affecting rates of re-suspension, although secondary nitrate and sulphate were also present in the larger particles.Concentrations of all measured species except sodium and chloride were higher during the burning season. Although concentrations were lower than often found in polluted urban environments, the massive increases during much of the year, due to a single anthropogenic activity (sugar cane burning) are indicative of a very large perturbation of the lower troposphere in the region relative to the natural condition. These aerosols are suspected of promoting respiratory disease. They also represent an important mechanism for the tropospheric transport of species relevant to surface acidification (sulphates, nitrates, ammonium and organic acids) and soil nutrient status (potassium, nitrogen, ammonium, calcium), so their impact on fragile natural ecosystems (following deposition) needs to be considered. (C) 2004 Elsevier Ltd. All rights reserved.

The sugar cane vegetal residues unloaded in the sugar mill: Operational costs and physical characteristics

Brazil produced in 2002/03 season 317.87×106tons of sugar cane stalks and 36.88×106tons of vegetal residues (green leaves, dry leaves and tops) in a planted area of 4.61×106 hectares (ha). These residues have a useful heat of 3,613.14Mcal.t-1. Currently most of this biomass is burned as a pre-harvest practice. The doubt persists in the system type that it must be adopted to pick up, load, transport and unload this biomass at the sugar mill boilers. This study analyzed 22 variables related to operational costs and physical characteristics of these residues in a field situation using a JOHN DEERE® 6850 forage harvester with two different treatments: T1 and T2 (two types of rakes) with 6 repetitions each one. The geographic location of the studied area that belongs to COSTA PINTO MILL (COSAN® Group) is: Latitude 22°40'30S and Longitude 47°36'38W. The adopted methodology was proposed by Ripoli et al. (2002). The obtained results at a 5% level of significance showed that both treatments did not differed significantly between them. Some of the results were, where EBP stands for Oil Equivalent Barrel: Windrowing (T1=US$0.17.EBP-1 and US$9.59.ha-1, T2=US$0.08.EBP-1 and US$4.27.ha-1); Pick up (T1=US$1.31.EBP-1 and US$44.29.ha-1, T2 =US$1.37.EBP-1 and US$48.36.ha-1); Transportation (T1=US$1.27.EBP-1 and US$14,30.ha -1, T2=US$1.33.EBP-1 and US$14,80.ha -1), Unloading at the sugar mill (T1=US$0.30.EBP-1 and US$3.39.ha-1, T2=US$0.32.EBP-1 and US$3.51.ha-1); Total (T1=US$3.05.EBP-1 and US$71.57.ha-1, T2=US$3.10.EBP-1 and US$70.94.ha-1). Confronting the obtained data with the ones in the bibliography, this system revealed itself more expensive than the baling system or the integral harvest system using combines.

Emissions from sugar cane fires in the Central & Western State of São Paulo and aerosol layers over metropolitan São Paulo observed by IPEN's lidar: Is there a connection?

The central and western parts of the State of São Paulo are well-known for vast sugar cane plantations, which during the harvest time are traditionally burnt about 12 hours before manual cutting. This procedure causes the release of large quantities of aerosols and a variety of gases, which can be observed by IPMet's radars, located in Bauru and Presidente Prudente, on days with no or little rain. Depending on the distance of these plumes from the radar, they can be detected up to 5 km amsl or more, and are subsequently being transported by winds to other regions. During the dry winter season of 2008, such plumes, attributed to cane fires, were frequently observed by IPMet's radars and documented in terms of radar reflectivity, time and location during the period 10 th - 21 st July 2008. At the same time, IPEN's Elastic Backscatter Lidar in São Paulo observed layers of aerosols of variable strength and heights above the city. The most significant days, viz. 14 and 15 July 2008 had been selected for calculating backward, as well as forward trajectories, deploying the European Flextra 3.3 Trajectory Model, which was initiated with ECMWF historic data with a 0,25 o×0,25 o grid spacing. The results presented here show an excellent match between the radar-detected sources of the plumes on 11 th July 2008 in the central parts of the State and the observations by IPEN's Lidar over Metropolitan São Paulo on 14 th July 2008, both in terms of forward and backward trajectories, as well as their heights, with a transport duration of approximately 70 hours under the prevailing meteorological conditions. © Sociedad Española de Óptica.