Tomato spotted wilt virus glycoproteins induce the formation of endoplasmic reticulum- and Golgi-derived pleomorphic membrane structures in plant cells

Tomato spotted wilt virus (TSWV) particles are spherical and enveloped, an uncommon feature among plant infecting viruses. Previous studies have shown that virus particle formation involves the enwrapment of ribonucleoproteins with viral glycoprotein containing Golgi stacks. In this study, the localization and behaviour of the viral glycoproteins Gn and Gc were analysed, upon transient expression in plant protoplasts. When separately expressed, Gc was solely observed in the endoplasmic reticulum (ER), whereas Gn was found both within the ER and Golgi membranes. Upon co-expression, both glycoproteins were found at ER-export sites and ultimately at the Golgi complex, confirming the ability of Gn to rescue Gc from the ER, possibly due to heterodimerization. Interestingly, both Gc and Gn were shown to induce the deformation of ER and Golgi membranes, respectively, also observed upon co-expression of the two glycoproteins. The behaviour of both glycoproteins within the plant cell and the phenomenon of membrane deformation are discussed in light of the natural process of viral infectio

Variabilidade genética de duas populações de Astyanax altiparanae da bacia do alto rio Paraná.

O presente trabalho tem como objetivo caracterizar a estrutura genética de duas populações de A. altiparanae da bacia do alto rio Paraná (Paranapanema e Tietê) através da análise de 11 marcadores moleculares do tipo microssatélite.Organizado por: Sílvio Ricardo Maurano; AQUACIÊNCIA 2012

Reprodução induzida e criação de larvas de híbridos interespecíficos e intragenérico da ordem siluriforme (Pisces).

O presente trabalho tem como objetivo analisar a resposta à indução artificial da reprodução em exemplares de cachapinta, híbrido resultante do cruzamento de Pseudoplatystoma corruscans X Pseudoplatystoma reticulatum no cruzamento intergenérico entre fêmea deste produto híbrido e macho de jundiá (Leiarius marmoratus), para verificar a produção de gametas viáveis, quantidade de larvas eclodidas, percentual de sobrevivência e crescimento das larvas e consequentemente a produção de exemplares para estudos genéticos.Organizado por: Sílvio Ricardo Maurano; AQUACIÊNCIA 2012

Urban land use of the Sao Paulo metropolitan area by automatic analysis of LANDSAT data

The separability of urban land use classes in the metropolitan area of Sao Paulo was studied by means of automatic analysis of MSS/LANDSAT digital data. The data were analyzed using the media K and MAXVER classification algorithms. The land use classes obtained were: CBD/vertical growth area, residential area, mixed area, industrial area, embankment area type 1, embankment area type 2, dense vegetation area and sparse vegetation area. The spectral analysis of representative samples of urban land use classes was done using the "Single Cell" analysis option. The classes CBD/vertical growth area, residential area and embankment area type 2 showed better spectral separability when compared to the other classes

Kinetic modeling and microbial assessment by fluorescent in situ hybridization in anaerobic sequencing batch biofilm reactors treating sulfate-rich wastewater

This paper reports the results of applying anaerobic sequencing batch biofilm reactors (AnSBBR) for treating sulfate-rich wastewater. The reactor was filled with polyurethane foam matrices or with eucalyptus charcoal, used as the support for biomass attachment. Synthetic wastewater was prepared with two ratios between chemical oxygen demand (COD) and sulfate concentration (COD/SO4(2-)) of 0.4 and 3.2. For a COD/SO4(2-) ratio of 3.2, the AnSBBR performance was influenced by the support material used; the average levels of organic matter removal were 67% and 81% in the reactors filled with polyurethane foam and charcoal, respectively, and both support materials were associated with similar levels of sulfate reduction (above 90%). In both reactors, sulfate-reducing bacteria (SRB) represented more than 65% of the bacterial community. The kinetic model indicated equilibrium between complete- and incomplete-oxidizing SRB in the reactor filled with polyurethane foam and predominantly incomplete-oxidizing SRB in the reactor filled with charcoal. Methanogenic activity seems to have been the determining factor to explain the better performance of the reactor filled with charcoal to remove organic matter at a COD/SO4(2-) ratio of 3.2. For a COD/SO4(2-) ratio of 0.4, low values of sulfate reduction (around 32%) and low reaction rates were observed as a result of the small SRB population (about 20% of the bacterial community). Although the support material did not affect overall performance for this condition, different degradation pathways were observed; incomplete oxidation of organic matter by SRB was the main kinetic pathway and methanogenesis was negligible in both reactors.This work was funded by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and the Financiadora de Estudos e Projetos (FINEP), Brazil. The authors acknowledge the grants received from FAPESP and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brazi

Endothelial heme oxygenase-1 induction by hypoxia. Modulation by inducible nitric-oxide synthase and S-nitrosothiols.

The stress protein heme oxygenase-1 (HO-1) is induced in endothelial cells exposed to nitric oxide (NO)-releasing agents, and this process is finely modulated by thiols (Foresti, R., Clark, J. E., Green, C. J., and Motterlini R. (1997) J. Biol. Chem. 272, 18411–18417). Here, we report that up-regulation of HO-1 in aortic endothelial cells by severe hypoxic conditions (pO2 ≤ 2 mm Hg) is preceded by increased inducible NO synthase and NO synthase activity. This effect is accompanied by oxidation of intracellular glutathione and formation of S-nitrosothiols. Incubation of cells with a selective inhibitor of inducible NO synthase (S-(2-aminoethyl)-isothiourea) or a NO scavenger ([2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide]) significantly attenuated the increase in heme oxygenase activity caused by reduced oxygen availability. A series of antioxidant agents did not prevent the elevation in heme oxygenase activity by hypoxia; however, the precursor of glutathione synthesis and thiol donor,N-acetylcysteine, completely abolished HO-1 induction. We also found that the hypoxia-mediated increase in endothelial heme oxygenase activity was potentiated by the presence ofS-nitrosoglutathione. These results indicate that intracellular interaction of thiols with NO is an important determinant in the mechanism leading to HO-1 induction by reduced oxygen levels. We suggest that in addition to oxidative stress, HO-1 gene expression can be regulated by redox reactions involving NO andS-nitrosothiols (nitrosative stress), emphasizing a versatile role for the heme oxygenase pathway in the cellular adaptation to a variety of stressful conditions

Effect of ammonia load on efficiency of nitrogen removal in an SBBR with liquid-phase circulation

The removal of biological nitrogen from a synthetic wastewater with different ammonium nitrogen concentrations (50 and 100 mgN-NH4+/L) by a nitrification and denitrification process using a sequencing batch biofilm reactor (SBBR) with liquid-phase circulation was studied. The system with a total working volume of 4.6 L (3.7 L in the reactor and 0.9 L in the reservoir) treated 2.1 L of synthetic wastewater in 12-h cycles. As inoculum two types of biomass were used: an anaerobic/anoxic one from an up-flow anaerobic sludge blanket reactor (UASB) and an aerobic one from a prolonged aeration activated sludge system. The system, maintained at 30 ± 1 ºC, operated in batch mode followed by fed-batch mode and was aerated intermittently. During fed-batch operation the reactor was fed with an external carbon source as electron donor in the denitrifying step and with no aeration. When the reactor was fed with 50 mgN-NH4+/L, efficiencies of removal of ammonium nitrogen and total nitrogen from the effluent were 93.8 and 72.2%, respectively, and nitrite, nitrate and organic nitrogen concentrations were 0.07, 6.4 and 0.5 mg/L, respectively. On the other hand, when the influent ammonium nitrogen concentration was 100 mgN-NH4+/L, residual nitrite and nitrate were 0.17 and 20.4, respectively, and no N-Org was found in the effluent. It should be mentioned that residual nitrate remained unaltered at the different C/N ratios used. Consequently, efficiency of total nitrogen removal was reduced to 66.7%, despite efficiency of ammonium nitrogen removal exceeding 90%. These results show the potential of the proposed system in removing ammonium nitrogen from liquid effluents with a moderate ammonium nitrogen concentration.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

A RAPD-PCR-based genetic diversity analysis of Helicoverpa armigera and H. zea populations in Brazil.

Therefore, the aim of this study was to determine the genetic diversity of H. armigera and H. zea populations by RAPD-PCR analysis. The most important result was the clustering of one H. armigera population in a group predominantly formed by H. zea. It could indicate a possible occurrence of an interspecific cross between these species. This is a concern to Brazilian agriculture due to the possibility of selection of hybrids well adapted to the American environment, which would be inherited from H. zea

Predominant Golgi-residency of the plant K/HDEL receptor is essential for its function in mediating ER retention

Accumulation of soluble proteins in the endoplasmic reticulum (ER) of plants is mediated by a receptor termed ER RETENTION DEFECTIVE 2 (ERD2) or K/HDEL receptor. Using two gain-of-function assays and by complementing loss of function in Nicotiana benthamiana we discovered that compromising the lumenal N-terminus or the cytosolic C-terminus with fluorescent fusions abolishes its biological function and profoundly affects its subcellular localization. Based on the confirmed asymmetrical topology of ERD2 we engineered a new fluorescent ERD2 fusion protein that retains biological activity. Using this fusion, we show that ERD2 is exclusively detected at the Golgi apparatus, unlike non-functional C-terminal fusions which also label the ER. Moreover, ERD2 is confined to early Golgi compartments and does not show ligand-induced redistribution to the ER. We show that the cytosolic C-terminus of ERD2 plays a crucial role in its function. Two conserved Leucine residues that do not correspond to any known targeting motifs for ER-Golgi trafficking were shown to be essential for both ERD2 Golgi residency and its ability to mediate ER retention of soluble ligands. The results suggest that anterograde ER to Golgi transport of ERD2 is either extremely fast, well in excess of the bulk flow rate, or that ERD2 does not recycle in the way originally proposed

A holographic system for subsea recording and analysis of plankton and other marine particles

We report here details of the design, development, initial testing and field-deployment of the HOLOMAR system for in-situ subsea holography and analysis of marine plankton and nonliving particles. HOLOMAR comprises a submersible holographic camera ("HoloCam") able to record in-line and off-axis holograms at depths down to 100 m, together with specialised reconstruction hardware ("HoloScan") linked to custom image processing and classification software. The HoloCam consists of a laser and power supply, holographic recording optics and holographic plate holders, a water-tight housing and a support frame. It utilises two basic holographic geometries, in-line and off-axis such that a wide range of species, sizes and concentrations can be recorded. After holograms have been recorded and processed they are reconstructed in full three-dimensional detail in air in a dedicated replay facility. A computer-controlled microscope, using video cameras to record the image at a given depth, is used to digitise the scene. Specially written software extracts a binarised image of an object in its true focal plane and is classified using a neural network. The HoloCam was deployed on two separate cruises in a Scottish sea loch (Loch Etive) to a depth of 100 m and over 300 holograms were recorded