Fat body cells of Amblyomma cajennense partially engorged females (Acari: Ixodidae) and their role on vitellogenesis process

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ultrastructural study of the ovary of the sugarcane spittlebug Mahanarva fimbriolata (Hemiptera)

The present study describes the ultrastructure of meroistic telotrophic ovaries of the sugarcane spittlebug Mahanarva fimbriolata. In this type of ovary, nurse cells, oogonia, and prefollicular tissue are located at the terminal (distal) regions or tropharium of ovarioles. Oocytes in different developmental stages, classified from I to V, are observed in the vitellarium. Stage I oocytes do not exhibit intercellular spaces in the follicular epithelium, suggesting that synthesis and production of yolk during this stage occurs only through endogenous processes. Small yolk granules of different electron densities are present in the cytoplasm. Few lipid droplets are observed. Stage 11 oocytes exhibit small intercellular spaces in the follicular epithelium. More protein as well as lipid yolk granules are observed in the cytoplasm. In stage III oocytes, intercellular spaces in the follicular epithelium are larger than those observed in the previous stage. Electrondense protein granules of various sizes, larger than those observed in stage 11 oocytes predominate in the cytoplasm. Smaller lipid droplets are also present. In stage IV oocytes, the follicular epithelium exhibits large intercellular spaces. Our data clearly indicate that the opening of these spaces in the follicular epithelium of M. fimbriolata oocytes increases as the intake of exogenous proteins intensifies, that is, in stages IV and Voocytes. During these stages, granular yolk becomes viscous due to the lysis of granules. In stage Voocytes, viscous yolk predominates in the cytoplasm. This type of yolk, however, has not been described for other orders of insects. The chorion of M. fimbriolata oocytes consists of an external layer (exochorion) and an internal one (endochorion), which is in direct contact with the oocyte. Numerous small pores that probably facilitate oxygenation of the internal structures inside the eggs are observed in the exochorion. (c) 2006 Elsevier Ltd. All rights reserved.

Female reproductive system of the sugarcane spittlebug Mahanarva fimbriolata (Auchenorrhyncha): Vitellogenesis dynamics and protein quantification

The present study aimed describing the ovaries of the sugarcane spittlebug Mahanarva fimbriolata which are meroistic telotrophic with nurse cells and oocytes located in the tropharium. SEM revealed paired ovaries located dorsolaterally around the intestine, and oocytes exhibiting shapes ranging from round (less developed) to elliptic (more developed), suggesting a simultaneous, although, asynchronous development. Based on histological data we classified the oocytes in stages from I to V. Stage I oocytes exhibit follicular epithelium with cubic and/or prismatic cells, fine cytoplasmic granules. Stage II oocytes present intercellular spaces in the follicular epithelium due to the incorporation of yolk elements from the hemolymph. Small granules are present in the periphery of oocytes while larger granules are observed in the center. Stage III oocytes are larger and intercellular spaces in the follicular epithelium are evident, as well as the interface between follicular epithelium and oocyte. Yolk granules of different sizes are present in the cytoplasm. During this stage, chorion deposition initiates. Stage IV oocytes exhibit squamous follicular cells and larger intercellular spaces when compared to those observed in the previous stage. The oocyte cytoplasm present granular and viscous yolk, the latter is the result of the breakdown of granules. Stage V oocytes exhibit a follicular epithelium almost completely degenerated, smaller quantities of granular yolk and large amounts of viscous yolk. Based on our findings we established the sequence of yolk deposition in M. fimbriolata oocyte as follows: proteins and lipids, which are first produced by endogenous processes in stages I and II oocytes. Exogenous incorporation begins in stage III. In stages I and II oocytes, lipids are also produced by follicular epithelial cells. The third element to be deposited is polysaccharides, mainly found as complexes. Therefore, the yolk present in the oocytes of this species consists of glycolipoproteins. Molecular weights of proteins present in M. fimbriolata oocytes ranged from 10 to 92 KDa, differently from vitellogenin, the most common protein present in insect oocytes, weighing approximately 180 KDa. (c) 2006 Elsevier Ltd. All rights reserved.

Interaction of human neutrophils with endothelial cells regulates the expression of endogenous proteins annexin 1, galectin-1 and galectin-3

Annexin 1 (ANXA1), galectin-1 (Gal-1) and galectin-3 (Gal-3) proteins have been identified as important mediators that promote or inhibit leukocyte migration. The expression of these proteins was studied in human neutrophils and endothelial cells (ECs) during a transmigration process induced by IL-8. Upon neutrophil adhesion to EC, a significant increase in the cleaved ANXA1 (LCS3, raised against all ANXA1 isoforms) expression was detected in the plasma membrane of adhered neutrophils and ECs compared to intact ANXA1 isoform (LCPS1, against N-terminus of protein). Adherent neutrophils had elevated Gal-3 levels in the nucleus and cytoplasm, and ECs in their plasma membranes. In contrast, a decrease in the total amounts of Gal-1 was detected in migrated compared to non-migrated neutrophils. Therefore, ANXA1 and Gal-3 changed in their content and localization when neutrophils adhere to endothelia, suggesting a process of sensitive-balance between two endogenous anti- and pro-inflammatory mediators. (c) 2006 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

Subcellular localization of proteins labeled with GFP in Xanthomonas citri ssp citri: targeting the division septum

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

Chromatoid body: Remnants of nucleolar proteins during spermatogenesis in triatomine (Heteroptera, Triatominae)

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

High-Fat Diet Obesity Associated With Insulin Resistance Increases Cell Proliferation, Estrogen Receptor, and PI3K Proteins in Rat Ventral Prostate

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

p53, p16 and Fhit Proteins Expressions in Chronic Esophagitis and Chagas Disease

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

The Venomics of Bothrops alternatus is a Pool of Acidic Proteins with Predominant Hemorrhagic and Coagulopathic Activities

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

Photoacoustic spectroscopy of aromatic amino acids in proteins

This paper concerns the use of photoacoustic spectroscopy (PAS) to study the presence of aromatic amino acid in proteins. We examined the aromatic amino acids in six proteins with well-known structures using absorption spectra of near ultraviolet PAS over the wavelength range 240-320 nm. The fundamental understanding of the physical and chemical properties that govern the absorption of light and a subsequent release of heat to generate a transient pressure wave was used to test the concept of monitoring aromatic amino acids with this method. Second derivative spectroscopy in the ultraviolet region of proteins was also used to study the regions surrounding the aromatics and the percentage area in each band was related in order to determine the contribution in function of the respective molar extinction coefficients for each residue. Further investigation was conducted into the interaction between sodium dodecyl sulphate (SDS) and bothropstoxin-I (BthTx-I), with the purpose of identifying the aromatics that participate in the interaction. The clear changes in the second derivative and curve-fitting procedures suggest that initial SDS binding to the tryptophan located in the dimer interface and above 10 SDS an increased intensity between 260 and 320 nm, demonstrating that the more widespread tyrosine and phenylalanine residues contribute to the SDS/BthTx-I interactions. These results demonstrate the potential of near UV-PAS for the investigation of membrane proteins/detergent complexes in which light scattering is significant.

The membranotropic activity of N-terminal peptides from the pore-forming proteins sticholysin I and II is modulated by hydrophobic and electrostatic interactions as well as lipid composition

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A systematic approach to identify STRE-binding proteins of the gsn glycogen synthase gene promoter in Neurospora crassa

The gene encoding glycogen synthase in Neurospora crassa (gsn) is transcriptionally down-regulated when mycelium is exposed to a heat shock from 30 to 45 degrees C. The gsn promoter has one stress response element (STRE) motif that is specifically bound by heat shock activated nuclear proteins. In this work, we used biochemical approaches together with mass spectrometric analysis to identify the proteins that bind to the STRE motif and could participate in the gsn transcription regulation during heat shock. Crude nuclear extract of heat-shocked mycelium was prepared and fractionated by affinity chromatography. The fractions exhibiting DNA-binding activity were identified by electrophoretic mobility shift assay (EMSA) using as probe a DNA fragment containing the STRE motif DNA-protein binding activity was confirmed by Southwestern analysis. The molecular mass (MM) of proteins was estimated by fractionating the crude nuclear extract by SDS-PAGE followed by EMSA analysis of the proteins corresponding to different MM intervals. Binding activity was detected at the 30-50 MM kDa interval. Fractionation of the crude nuclear proteins by IEF followed by EMSA analysis led to the identification of two active fractions belonging to the pIs intervals 3.54-4.08 and 6.77-7.31. The proteins comprising the MM and pI intervals previously identified were excised from a 2-DE gel, and subjected to mass spectrometric analysis (MALDI-TOF/TOF) after tryptic digestion. The proteins were identified by search against the MIPS and MIT N. crassa databases and five promising candidates were identified. Their structural characteristics and putative roles in the gsn transcription regulation are discussed.

Differential proteomic analysis of Acidithiobacillus ferrooxidans cells maintained in contact with bornite or chalcopyrite: Proteins involved with the early bacterial response

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Study of the proteins in the defatted flour and protein concentrate of baru nuts (Dipteryx alata Vog)

O baru (Dipteryx alata Vog.) é uma leguminosa abundante no Cerrado brasileiro, cuja castanha pode ser explorada através do uso sustentável para o aproveitamento das frações proteicas e lipídicas. Este trabalho teve como objetivo estudar as proteínas desta castanha, presentes na farinha desengordurada e no concentrado proteico, quanto as suas propriedades funcionais, ao perfil das frações proteicas e à digestibilidade in vitro. A farinha desengordurada com hexano foi submetida à extração no pH de maior solubilidade das proteínas, obtendo-se o concentrado proteico. O perfil eletroforético das frações proteicas foi avaliado em gel de SDS-PAGE. As propriedades funcionais indicaram a possibilidade de emprego em diversos alimentos, assim como a soja, conferindo capacidade de absorção de água, capacidade de absorção de óleo, propriedades emulsificantes e espumabilidade. As globulinas, seguidas das albuminas, são as frações majoritárias da farinha e do concentrado proteico, respectivamente. A digestibilidade foi superior no concentrado em relação à farinha desengordurada.

Dynamics and cytochemistry of oogenesis in Astyanax fasciatus (Cuvier) (Teleostei, Characiformes, Characidae) from Rio Sapucaí, Minas Gerais State, Brazil

Oogenesis involves a set of transformations which are undergone by female germ cells These cells change into oogonias and then into mature oocytes. Sexually mature female fish were collected monthly, during one year, from the Sapucaí River, a tributary of the Rio Grande, which is part of the Furnas Reservoir in the state of Minas Gerais. During the several stages of maturation, we observed small round oogonias with a large nucleus, a single nucleolus, and weakly stained cytoplasm with eosinophilic granules. The primary oocytes showed a large basophilic nucleus, with a developed peripheral nucleolus and a reduced cytoplasm. The previtellogenic oocytes presented voluminous cytoplasm and nucleus with several small peripheral nucleoli. The oocytes underwent vitellogenesis with the development of the zona radiata and the follicle cells. Their cytochemical reactions indicated that the two layers of the zona radiata of A. fasciatus contained proteins and polysaccharides. The initially squamous follicle cells, became cuboidal. In mature oocytes, the nucleus moved toward the periphery, next to the micropyle, and the yolk granules formed by proteins, fulfilled the cytoplasm. The clear unstained vesicles are likely to be the cortical alveoli in the perivitelline region.