Avaliação das estruturas de cobertura em madeira de um galpão de estoque de produtos químicos

Este trabalho tem com objetivo avaliar a integridade das estruturas que fazem parte da cobertura de um galpão de 70 anos, destinado ao estoque de produtos químicos, localizada no Estado de São Paulo (Brasil), e apontar possíveis soluções para os problemas identificados. Buscou-se estimar a influência dos resíduos dos produtos químicos depositados nas superfícies dos elementos estruturais e identificar a espécie de madeira utilizada no projeto, por meio de ensaios físicos (densidade aparente) e mecânicos (compressão e cisalhamento paralelo às fibras. Os problemas identificados foram: (a) umidade devido a telhas quebradas; (b) deterioração superficial da madeira em função de reação química entre os produtos armazenados e a estrutura; (c) cavilhas de madeira com instalação incompleta; (d) falta de mãos-francesas em algumas terças; (e) travamento inadequado das treliças; e (f) deterioração de pregos de fixação por oxidação. Pelas observações realizadas, as soluções indicadas são de fácil execução, pois as estruturas não foram severamente prejudicadas. Com relação à espécie de madeira, os resultados das propriedades mecânicas obtidas associadas à densidade aparente apontaram para a espécie Peroba-Rosa (Aspidosperma polyneuron). A análise estatística de resultados apontou para a não influência do produto químico no desempenho da estrutura do telhado

Intermediate Tyrosyl Radical and Amyloid Structure in Peroxide-Activated Cytoglobin.

We characterized the peroxidase mechanism of recombinant rat brain cytoglobin (Cygb) challenged by hydrogen peroxide, tert-butylhydroperoxide and by cumene hydroperoxide. The peroxidase mechanism of Cygb is similar to that of myoglobin. Cygb challenged by hydrogen peroxide is converted to a Fe4+ oxoferryl π cation, which is converted to Fe4+ oxoferryl and tyrosyl radical detected by direct continuous wave-electron paramagnetic resonance and by 3,5-dibromo-4-nitrosobenzene sulfonate spin trapping. When organic peroxides are used as substrates at initial reaction times, and given an excess of peroxide present, the EPR signals of the corresponding peroxyl radicals precede those of the direct tyrosyl radical. This result is consistent with the use of peroxide as a reducing agent for the recycling of Cygb high-valence species. Furthermore, we found that the Cygb oxidation by peroxides leads to the formation of amyloid fibrils. This result suggests that Cygb possibly participates in the development of degenerative diseases; our findings also support the possible biological role of Cygb related to peroxidase activity

Changes in the EA spectrum of Cygb during the reaction with <i>t</i>-BuOOH.

<p>A) Bleaching of Soret and Q bands of EA spectra of Cygb in the course of the reaction with <i>t</i>-BuOOH. The black line represents the EA spectrum of resting Cygb, red, green and blue lines corresponds to the spectra obtained at 30, 60 and 200 s after addition of <i>t</i>-BuOOH and indicated by the arrows. B) Normalized spectra of Cygb resting form and 200 s after <i>t</i>-BuOOH addition. C) Differential spectra of Cygb obtained 30 and 200 s after the addition of <i>t</i>-BuOOH. The experiments of EA spectroscopy were performed using 65 μmol.L^-1 Cygb and 0.1 cm optical length. When present, the concentration of peroxides was 650 μmol.L^-1. These results are representative of three independent replicates.</p

Formation of Cygb amyloid structure after challenge by peroxides.

<p>A), B) and C) show, respectively the epifluorescence images of Cygb control, control plus GSH and challenged by hydrogen peroxide obtained immediately (left panels) 24 h (right panels) after incubation and staining by thioflavine-T. For the low-vacuum SEM experiments, it was used 7 μmol.L^-1 cygb solution with 70 μmol.L^-1 peroxide solutions. For the epifluorescence experiments 70 μmol.L^-1 protein solution was incubated for 1 h with 700 μmol.L^-1 peroxide solution in the presence of thioflavin-T. For FTIR measurements, 7 μmol.L^-1 protein solution was incubated with 70 μmol.L^-1 peroxide solutions for 1 h. The results are representative of three independent experiments.</p

Changes in the EA spectrum of Cygb during the reaction with hydrogen peroxide.

<p>A) Bleaching of Soret and Q bands of EA spectra of Cygb in the course of the reaction with hydrogen peroxide. The black line represents the EA spectrum of resting Cygb, red, green and blue lines corresponds to the spectra obtained at 30, 60 and 200 s after addition of hydrogen peroxide and indicated by the arrows. B) Normalized spectra of Cygb resting form and 200 s after hydrogen peroxide addition. C) Differential spectra of Cygb obtained 30 and 200 s after the addition of hydrogen peroxide The experiments of EA spectroscopy were performed using 65 μmol.L^-1 Cygb and 0.1 cm optical length. When present, the concentration of peroxide was 650 μmol.L^-1. These results are representative of three independent replicates.</p

Interatoma of rat Cygb with hydrogen peroxide.

<p>The network shows, in each node, a protein predicted to have functional links with Cygb and hydrogen peroxide. Inside the figure the abbreviations are SOD1 (superoxide dismutase [Cu-Zn]), Hmox2 (heme oxygenase 2 [HO-2]), Mb (myoglobin), Mpo (myeloperoxidase), cat (catalase), Cygb (cytoglobin), Prdx1 (peroxyredoxin-1), Prdx5 (peroxyredoxin-5) and Srxn1 (Ab2-390). In the figure light green, cyan and magenta lines correspond, respectively, to textmining, databases and experiments supporting the relationship among the proteins and hydrogen peroxide.</p

EPR spectrum of resting Cygb and the spectral components obtained by simulation.

<p>The red line corresponds to simulation of the composite spectrum and the blue and green lines correspond to the high and low spin components, respectively. The g values of the low spin state component are: g1 = 3.228, g2 = 2.033 and g3 = 1.385 with rhombic distortion and for the high spin state the g values are: g1 = 6.062, g2 = 5.785 and g3 = 2.0409. The simulation was done by the software Symphonia. For EPR experiments, the protein concentration was of 1.2 mmol.L^-1. These results are representative of three independent replicates.</p

Spectroscopic characteristics of Cygb.

<p>The upper panel shows the EA spectrum of Cygb and the respective inset the corresponding far-UV CD spectrum. The lower panel shows the CD (light gray line) and the MCD spectra of Cygb obtained by addition and subtraction of the original spectra obtained at positive and negative magnetic fields. MCD is shown at increasing magnetic fields and the respective inset shows the linear increase of Soret band intensity promoted by increasing the magnetic field. The experiments of EA spectroscopy were performed with 65 μmol.L^-1 Cygb using 0.1 cm optical length. The experiments of CD and MCD were performed using 20 μmol.L^-1 protein solution in 20 mmol.L^-1 phosphate buffer, pH 7.4. These results are representative of three independent replicates.</p

Changes in the EPR spectrum of resting Cygb during the reaction with hydrogen peroxide.

<p>The spectra marked as a, b and c were obtained at 30, 60 and 210 s after the addition of hydrogen peroxide. The inset shows a zoom in the spectra of the free radical produced concomitantly with the formation of high valence species. For EPR experiments, the protein concentration was of 1.2 mmol.L^-1and when present, the peroxide concentration was of 12 mmol.L^-1. These results are representative of three independent replicates.</p

Leviserpin: A Serine Peptidase Inhibitor (Serpin) from the Sugarcane Weevil Sphenophorus levis

Serine peptidase inhibitors (serpins) form a superfamily of proteins covering abroad spectrum of different biological functions. Here we describe the inhibitory characterization of leviserpin, the first serpin from the sugar cane weevil Sphenophorus levis. Leviserpin was able to inhibit bovine trypsin by the formation of the covalent complex serpin-peptidase, demonstrated by SDS-PAGE and mass spectroscopy analysis. We also have determined the cleavage site at the reactive center loop, by the analysis of the polypeptides released from de C-terminus of leviserpin. Moreover we investigated the mRNA expression of leviserpin in different stages of S. levis development. Thus the specificity of leviserpin, in addition with its mRNA coding being transcribed through all lifecycle of the insect, can suggest a possible role in defense mechanism by regulating the action of prophenoloxidase (proPO) activating enzyme.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fed Univ ABC, Ctr Nat & Human Sci, Biol Chem Lab, BR-09210170 Santo Andre, SP, BrazilUniv Fed Sao Carlos, Mol Biol Lab, Dept Genet & Evolut, BR-13565905 Sao Carlos, SP, BrazilUniversidade Federal de São Paulo, Dept Biophys, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biophys, São Paulo, BrazilFAPESP: 98/14138-2FAPESP: 06/53607-6CNPq: 312701/2009-8Web of Scienc