Repositório RCAAP

Background: The magnetic albumin nanosphere (MAN), encapsulating maghemite nanoparticles, was designed as a magnetic drug delivery system (MDDS) able to perform a variety of biomedical applications. It is noteworthy that MAN was efficient in treating Ehrlich’s tumors by the magnetohyperthermia procedure. Methods and materials: In this study, several nanotoxicity tests were systematically carried out in mice from 30 minutes until 30 days after MAN injection to investigate their biocompatibility status. Cytometry analysis, viability tests, micronucleus assay, and histological analysis were performed. Results: Cytometry analysis and viability tests revealed MAN promotes only slight and temporary alterations in the frequency of both leukocyte populations and viable peritoneal cells, respectively. Micronucleus assay showed absolutely no genotoxicity or cytotoxicity effects and histological analysis showed no alterations or even nanoparticle clusters in several investigated organs but, interestingly, revealed the presence of MAN clusters in the central nervous system (CNS). Conclusion: The results showed that MAN has desirable in vivo biocompatibility, presenting potential for use as a MDDS, especially in CNS disease therapy.

O artigo analisa características do Estado desenvolvimentista no período pós-neoliberal no Brasil e na Argentina. A emergência de uma fase pós-neoliberal na América Latina está associada, ainda que sob diferentes matizes ideológicas, à ideia de expansão de políticas sociais e à retomada do papel do Estado na formulação e incentivo a estratégias de desenvolvimento econômico. Um aspecto fundamental para a eficácia das estratégias de desenvolvimento são as instituições e organizações envolvidas politica e economicamente. Como mostra a literatura, o desafio de construir um Estado desenvolvimentista requer a combinação de um Estado facilitador com uma estratégia de upgrading industrial acompanhada de aumento da inclusão social. Uma diferença relevante no caso brasileiro em relação ao caso argentino foi a permanência de instituições que foram construídas durante o período desenvolvimentista, com especial destaque para o BNDES. Ao verificar a relação entre Estado e desenvolvimento nos dois países, foi possível identificar quatro diferenças fundamentais do Brasil em relação à Argentina; uma forte instituição financeira de desenvolvimento e instituições financeiras públicas, uma maior capacidade estatal com uma burocracia estável e meritocrática no caso brasileiro, políticas sociais de longo prazo e uma relação mais cooperativa entre Estado e empresariado. _______________________________________________________________________________________ ABSTRACT

A gene encoding the entire highly expressed protein previously identified in the proteome of Paracoccidioides brasiliensis yeast cells as PbY20 has been isolated. The pby20 sequence reveals an open reading frame of 1364 bp and a deduced amino acid sequence of 203 residues, which shows high identity to benzoquinone reductase from Phanerochaete chrysosporium (72.0%), Saccharomyces cerevisiae Ycp4 (65%), and Schizosaccharomyces pombe p25 (59%), and to allergens from Alternaria alternata Alt a7 (70%) and from Cladosporium herbarum, Cla h5 (68%). Low levels of the pby20 transcript in the mycelium and highly induced ones in infective yeast cells during the transition of this dimorphic fungus indicate transcriptional control of its expression. PbY20 was immunologically detected only in yeast cell extract, suggesting an important role in cell differentiation or even in the maintenance of the yeast form. Immunoelectron microscopy showed that PbY20 is found inside large granules and vacuoles, in the nucleus, and also in the cytoplasm. Through sequence comparisons analysis and fluorescence emission assay, PbY20 was recognized as a member of the flavin mononucleotide flavodoxin-like WrbA family, which are involved in heat shock and oxidative stress in biological systems. Assuming that PbY20 belongs to this family, a similar role could be attributed to this protein.

O Censo e a pesquisa amostral nos projetos de reforma agrária originaram-se de uma demanda do Incra/Ministério de Política Fundiária em agosto de 1996, tendo em vista a necessidade de identificar todas as famílias assentadas em áreas de projetos de reforma agrária do Governo Federal, bem como traçar o perfil sócio-econômico das mesmas. A controvérsia sobre o número efetivo de famílias assentadas sempre representou um debate vazio devido à ausência de informações objetivas e atualizadas. Governo e organizações/movimentos sociais vinham apresentando dados discrepantes sobre o assunto, principalmente sobre as famílias efetivamente assentadas e suas condições reais de vida. Além disso, o Incra identificou a necessidade de atualizar seu cadastro de famílias assentadas, defasado em muitos estados. Em função da premência do levantamento de dados e da agilidade exigida percebeu-se que o quadro de técnicos do Incra era insuficiente para atender tal demanda. O envolvimento das universidades brasileiras ocorreu a partir de um debate já iniciado e coordenado pelo Forum de Reforma Agrária, realizado na Universidade de Brasília (UnB) nos meses de junho e julho de 1996. As universidades representadas naquele Forum apresentaram seu interesse em participar mais ativamente e se dispuseram a atender as demandas do Ministério Extraordinário de Política Fundiária, principalmente no que se referia a pesquisas. Desse modo, a participação das universidades no projeto do Censo e Pesquisa Amostral se deu através do Conselho de Reitores das Universidades Brasileiras (Crub), colocando professores e estudantes à disposição do projeto. As universidades brasileiras, ao defenderem sua participação, destacavam a importância de envolver estudantes e professores nas atividades mais práticas e realistas do cotidiano do processo de reforma agrária, apostando no efeito multiplicador da experiência tanto em termos de formação de uma cultura a respeito do assunto, como no fortalecimento do debate teórico sobre a reforma agrária. A Universidade de Brasília foi convidada pelo Incra para coordenar o projeto, definindo a metodologia em conjunto com seus técnicos, assumir a tarefa de administrar os recursos, bem como designar os supervisores estaduais a partir de um perfil de pesquisador/professor em questões agrárias e possibilitar todas as condições técnicas e materiais ao desenvolvimento do Projeto. Participaram, efetivamente, 29 universidades entre federais, estaduais e comunitárias, 41 professores/ supervisores e 1.800 estudantes.

Power, the central notion in this text, has many definitions. My own conception is based on a combination of three different sources. For Richard Adams (1967), power is the control that one party posseses over another party’s environment. Of the several visions of Max Weber, I will retain that of power as the capacity to make people do things they do not want to do. Eric Wolf’s (1999) notion of structural power underscores the capacity historical relationships and forces—especially those that define access to social labour—have to create and organize settings that constrain people’s possibilities for action, and to specify the direction and distribution of energy flows. Power, thus, is about (a) to be the subject of one’s own environment, to be able to control one’s own destiny, i.e., the course of action or events that will keep one’s life as it is or will modify it, or (b) to prevent people from becoming such empowered actors. Since development is always about transformation (Berman, 1987), and typically occurs through encounters between insiders and outsiders located in different power positions, ownership of development initiatives is anchored in and influenced by situations where power inequalities abound. The difficulty of implementing change within the development community is intimately related to the fact that it is a power field.

Este artigo analisa os resultados do primeiro e segundo turnos da eleição municipal de 2000 no Brasil. Enfoca o desempenho de cada um dos principais partidos e de alguns dos principais candidatos, nas regiões do país e nas principais cidades, analisa o desempenho geral das candidatas mulheres, e procura traçar considerações sobre o impacto dos resultados no cenário político das eleições gerais de 2002. _______________________________________________________________________________________ ABSTRACT

The objective was to determine the morphological and ultrastructural features of sheep primordial follicles preserved in either 0.9% saline solution or TCM 199 at different temperatures. Soon after death, the ovarian pair of each ewe (n=5) was divided into 25 fragments. One fragment was immediately fixed for morphological evaluation (control). The other 24 fragments were randomly distributed in tubes containing 2 ml of 0.9% saline solution or TCM 199 and maintained at 4, 20 or 39 °C for 2, 4, 12, or 24 h. Based on histological assessment, storage of ovarian fragments in 0.9% saline solution at 20 °C for up to 24 h and in both solutions at 39 °C for 4, 12 or 24 h increased (P<0.01) the percentage of degenerate primordial follicles compared with controls. In contrast, preservation at 4 °C in both solutions, kept the percentage of morphologically normal primordial follicles similar to control values. Although histological integrity of primordial follicles was maintained in fragments stored at 20 °C for up to 24 h in TCM 199, these results were not confirmed by ultrastructural analysis. Based on transmission electron microscopy, only primordial follicles stored at 4 °C for up to 24 h, at 20 °C for up to 12 h and at 39 °C for up to 2 h in both solutions were ultrastructurally normal. In conclusion, sheep primordial follicles were successfully preserved at 4 °C for up to 24 h, at 20 °C for up to 12 h and at 39 °C for 2 h in 0.9% saline solution or TCM 199.

Este projeto faz parte de uma antropologia crítica da antropologia, uma perspectiva que descentraliza, re-historiciza, e pluraliza o que tem sido considerado “antropologia” até então. Ele questiona não só os conteúdos, mas também os termos e as condições dos encontros antropológicos. “Antropologias Mundiais” têm como objetivo a construção de um cânone policêntrico, que, de forma parecida ao multiculturalismo policêntrico (Shohat e Stam citado em Turner 1994), implica em uma reconceitualização dos relacionamentos entre comunidades antropológicas. A observação introdutória refere-se ao meu entendimento da antropologia como uma cosmopolítica. A noção de cosmopolítica procura prover uma perspectiva crítica e plural sobre as possibilidades de articulações supra e transnacionais. Ela é baseada, por um lado, nas evocações positivas associadas historicamente à noção de cosmopolitismo e, por outro lado, em análises nas quais assimetrias de poder são de fundamental importância (sobre cosmopolítica veja Cheah e Robbins 1998, e Ribeiro 2003).

In this study, the interaction of DMSA-coated magnetite nanoparticles (5 and 10 nm core-size) with Saccharomyces cerevisae was investigated using magnetic resonance (MR) and transmission electron microscopy (TEM). The TEM micrographs revealed magnetite nanoparticles attached externally to the cell wall. The MR data support the strong interaction among the nanoparticles supported by the cells. A remarkable shift in the resonance field was used as signature of particle attachment to the cell wall.

Sperm ultrastructure of five teiid lizards (Callopistes flavipunctatus, Crocodilurus amazonicus, Dicrodon guttulatum, Dracaena guianensis, and Teius oculatus), and the gymnophthalmid Cercosaura ocellata is described for the first time. Comparisons of sperm ultrastructure among these species and with those of previously examined teiids and gymnophthalmids revealed that the two groups of Teiioidea (Gymnophthalmidae and Teiidae), and the two subfamilies of Teiidae (Teiinae and Tupinambinae) could be distinguished on the basis of sperm ultrastructure data. Significant differences in sperm dimensions between Cnemidophorus and Aspidoscelis support the recent splitting of these two lineages into different genera. Our results revealed high levels of inter-generic variability in sperm ultrastructure within Teiidae, which produces a data set useful in analyzing relationships between genera and families. In phylogenetic analyses, however, sampling multiple species within teiid genera is essential and recording sperm measurements may profitably complement qualitative ultrastructural characters, maximizing the information content of these structures.

Freeze-drying sperm is an alternative to cryopreservation. Although sperm from various species has been freeze-dried, there are few reports for bovine sperm. The primary objective of this study was to evaluate the protective effect of various freeze-drying media on the structural and functional components of bovine sperm. The media tested were composed of TCM 199 with Hanks salts supplemented with 10% fetal calf serum (FCS) and TCM 199 with Hanks salts supplemented with 10% FCS and 0.2 M trehalose and EGTA solution. The efficiency of each medium on the preservation of freeze-dried sperm structures was evaluated with conventional and electron microscopy, DNA integrity was analyzed by a TUNEL assay, and fertilizing ability of lyophilized sperm was determined with ICSI. Although the plasma membrane was damaged in all media tested, mitochondria were similarly preserved in all freeze-drying treatments. The acrosome was best preserved in the media that contained trehalose (other treatments also conserved this structure). In contrast, media containing EGTA or trehalose most effectively preserved the nuclei in freeze-dried sperm, with only 2 and 5%, respectively, of cells with fragmented DNA. Furthermore, sperm conserved with these media also had higher (P < 0.05) rates of sperm head decondensation (32.5 and 27.5%), pronucleus formation (37.5 and 45.0%) and blastocyst formation (19.4 and 18.3%) than medium supplemented with FCS (15.0, 20.0 and 10.2%, respectively). In conclusion, media with EGTA and trehalose adequately protected bovine sperm during freeze-drying by preserving the viability of their nuclei.

The aim of this study was to verify the histological and ultrastructural characteristics of sheep preantral follicles after exposure of ovarian tissue to cryopreservation in glycerol (GLY), ethylene glycol (EG), propanediol (PROH) or dimethyl sulfoxide (DMSO) in order to determine the optimum method to store sheep ovarian tissue for later experimental or clinical use. Each ovarian pair from five mixed-breed ewes was divided into 17 fragments. One (control) fragment was immediately fixed for routine histological and ultrastructural studies and the remaining (test) fragments were randomly distributed in cryotubes, equilibrated at 20 °C/20 min in 1.8 mL of minimal essential medium (MEM) containing 1.5 or 3 M GLY, EG, PROH or DMSO and then either fixed for morphological studies to determine their possible toxic effect or frozen/thawed and then fixed to test the effect of cryopreservation on preantral follicles. Histological analysis showed that, compared to control fragments, all cryoprotectants at both concentrations significantly reduced the percentage of normal preantral follicles in ovarian fragments prior to or after cryopreservation. PROH 3.0 M appeared to exert a more toxic effect (P < 0.05) than the other cryoprotectants in noncryopreserved tissues. After freezing/thawing, the highest (P < 0.05) percentages of lightmicroscopical normal preantral follicles were observed in ovarian fragments cryopreserved in EG (1.5 and 3 M) or DMSO (1.5 M). However, transmission electronic microscopical (TEM) examination showed that only the DMSO-cryopreserved preantral follicles had normal ultrastructure. The data suggest that sheep preantral follicles should be cryopreserved with 1.5 M DMSO for later clinical or experimental application.

Tese (doutorado)—Universidade de Brasília, Faculdade de Tecnologia, Departamento de Engenharia Civil e Ambiental, 2008.

The spermatozoa of Chrysomya megacephala are similar to those described for other Brachycera. In this species, the spermatozoa are long and thin, measuring about 590 μm in length, of which the head region measures approximately 60 μm. The head includes a monolayered acrosome with electron-lucid material, and the shape of the nucleus, in cross-sections, varies from circular to oval with completely condensed chromatin. The centriole was observed in the zone of flagellar implantation, below the “peg” region. In the region of overlap, the followings structures are observed: nucleus, centriolar adjunct, mitochondrial derivatives and axoneme. The two mitochondrial derivatives are of different lengths but similar diameter. The axoneme is of a conventional insectan type with a 9 + 9 + 2 microtubular arrangement, with accessory tubules flanked by the electron-dense intertubular material. The male internal reproductive tract consists of testis, vas deferens, seminal vesicle, accessory glands and ejaculatory duct.

The objective of this study was to characterize the morphometry and ultrastructure of porcine preantral and antral follicles, especially the lipid component evolution. Ovarian tissue was processed for light microscopy. Ovarian tissue and dissected antral follicles (< 2, 2–4, and 4–6 mm) were also processed for transmission electron microscopy using routine methods and using an osmium-imidazole method for lipid detection. Primordial follicles (34 ± 5 μm in diameter, mean ± SD) had one layer of flattened-cuboidal granulosa cells around the oocyte, primary follicles (40 ± 7 μm) had a single layer of cuboidal granulosa cells around the oocyte, and secondary follicles (102 ± 58 μm) had two or more layers of cuboidal granulosa cells around the oocyte. Preantral follicle oocytes had many round mitochondria and both rough and smooth endoplasmic reticulum. In oocytes of primordial and primary follicles, lipid droplets were abundant and were mostly located at the cell poles. In secondary and antral follicles, the zona pellucida completely surrounded the oocyte, whereas some microvilli and granulosa cells projected through it. Numerous electron-lucent vesicles and vacuoles were present in the oolemma of secondary and antral follicles. Based on osmium-imidazole staining, most of these structures were shown to be lipid droplets. As the follicle developed, the appearance of the lipid droplets changed from small and black to large and gray, dark or dark with light streaks, suggesting that their nature may change over time. In summary, although porcine follicles and oocytes had many similarities to those of other mammalian species, they were rich in lipids, with lipid droplets with varying morphological patterns as the follicle developed.

Iron is essential for the proliferation of fungal pathogens during infection. The availability of iron is limited due to its association with host proteins. Fungal pathogens have evolved different mechanisms to acquire iron from host; however, little is known regarding how Paracoccidioides species incorporate and metabolize this ion. In this work, host iron sources that are used by Paracoccidioides spp. were investigated. Robust fungal growth in the presence of the iron-containing molecules hemin and hemoglobin was observed. Paracoccidioides spp. present hemolytic activity and have the ability to internalize a protoporphyrin ring. Using real-time PCR and nanoUPLC-MSE proteomic approaches, fungal growth in the presence of hemoglobin was shown to result in the positive regulation of transcripts that encode putative hemoglobin receptors, in addition to the induction of proteins that are required for amino acid metabolism and vacuolar protein degradation. In fact, one hemoglobin receptor ortholog, Rbt5, was identified as a surface GPI-anchored protein that recognized hemin, protoporphyrin and hemoglobin in vitro. Antisense RNA technology and Agrobacterium tumefaciensmediated transformation were used to generate mitotically stable Pbrbt5 mutants. The knockdown strain had a lower survival inside macrophages and in mouse spleen when compared with the parental strain, which suggested that Rbt5 could act as a virulence factor. In summary, our data indicate that Paracoccidioides spp. can use hemoglobin as an iron source most likely through receptor-mediated pathways that might be relevant for pathogenic mechanisms.

Cryopreservation of ovarian tissue may be a potential alternative for the conservation of genetically superior animals, including high milk- and meat-producing goat breeds. However, until now, no information was available concerning the cryopreservation of preantral follicles (PF) enclosed in caprine ovarian tissue. The objective of the present study was to evaluate the structural and ultrastructural characteristics of caprine PF after exposure to and cryopreservation of ovarian tissue in 1.5 and 3 M glycerol (GLY) and ethylene glycol (EG). At the slaughterhouse, each ovarian pair from five adult mixed breed goats was divided into nine fragments and randomly distributed into treatment groups. One fragment was immediately fixed for histological examination and ultrastructural analysis, after slaughter (control). Four of the ovarian fragments were equilibrated at 20 °C for 20 min in 1.8 ml of MEM containing 1.5 or 3 M GLY or EG for a toxicity test and the final four fragments were slowly frozen using these cryoprotectants at the concentrations above. After toxicity testing and freezing/thawing, the ovarian fragments were fixed for histological examination. Histological analysis showed that after toxicity testing and cryopreservation of the ovarian tissue in GLY or EG at both concentrations, the percentage of normal PF was significantly lower than controls. Ultrastructural analysis of PF frozen in 1.5 and 3 M GLY, as well as 3 M EG demonstrated that these follicles remained morphologically normal. In conclusion, we demonstrated cryopreservation of caprine PF in ovarian tissue.

Bovine preantral follicles within ovarian fragments were exposed and cryopreserved in absence or presence of 1.5 M glycerol (GLY), ethylene glycol (EG), propanediol (PROH) or dimethyl sulfoxide (DMSO), undergoing a previous cooling at 20 °C for 1 h (protocol 1) or at 4 °C for 24 h (protocol 2) in 0.9% saline solution. At the end of each treatment, preantral follicles were classified as non-viable/viable when they were stained/not stained with trypan blue, respectively. To confirm viability staining, ultra-structure of the follicles was evaluated by transmission electronic microscopy (TEM). Data were compared by Chi-square test (P < 0.05). The storage of the ovaries at 20 °C for 1 h (78%) and 4 °C for 24 h (80%) did not reduce significantly the percentage of viable preantral follicles when compared to the control (75%). Similar results were obtained when ovarian fragments, respectively, for protocols 1 and 2, were exposed to MEM (78 and 77%), 1.5 M EG (78 and 71%), as well as frozen in 1.5 M EG (74 and 77%). Percentages of viable follicles in control were similar to those observed after exposure (75%) and freezing (76%) in presence of 1.5 M DMSO only when protocol 1 was used. The increase of the concentration from 1.5 to 3.0 M, for all cryoprotectants, reduced significantly the percentage of viable preantral follicles after freezing. Ultra-structural analysis has confirmed trypan blue results, showing that not only basement membrane, but also organelles, were intact in viable preantral follicles. In conclusion, ovarian tissue cooling at 4 °C for 24 h before cryopreservation (protocol 2) does not affect the viability of bovine preantral follicles when 1.5 M EG is present in the cryopreservation medium.

The aim of this study was to investigate the effects of chronic exposure to soy isoflavones concentrate on the morphology of reproductive organs, semen quality, puberty age, serum levels of testosterone and sexual behavior of male rabbits. With this purpose, pregnant female rabbits were randomly assigned to receive orally 2.5mg (ISF 2.5) or 10mg (ISF 10) of soy isoflavones/kg of body wt/day. The animals of control group were manipulated as the other groups and received placebo (corn starch). All the rabbits were maintained on a soy-and alfafa-free diet throughout the gestation and lactation. Their male offspring received the same treatments from weaning to 33 weeks of age. Chronic exposure to isoflavones did not induce statistically significant alteration in the age at puberty, semen volume, daily sperm output, sperm concentration, motility, vigor and abnormalities. Also, isoflavones exposure had no effects on serum testosterone levels or sexual behavior in any group. Histopathologic evaluation did not reveal alterations in the testis, epididymis, prostate and pro-prostate glands of the rabbits. Taken together, these results show that gestational, lactational and post-lactational exposure to soy isoflavones, in doses comparable to those found in soy-containing animal and human diets, has no adverse effects on the reproductive parameters of male rabbits.

Cryopreservation of ovarian tissue is a new and promising technique for germ-line storage. The objective of this study was to evaluate the effect of four cryoprotectants (at two concentrations each) on the preservation of zebu bovine preantral follicles after ovarian cryostorage. Strips of ovarian cortex were cryopreserved using glycerol (GLY; 10 or 20%), ethylene glycol (EG), propanediol (PROH) or dimethylsulphoxide (DMSO; 1.5 or 3 M). In addition, a toxicity test was performed for each cryoprotectant by exposing the ovarian tissue to them without freezing. Tissues were analyzed by histology and transmission electron microscopy. Ovarian tissue frozen in either concentration of DMSO or PROH or in 10% GLY retained a higher percentage of morphologically normal follicles (73–88%) than tissue frozen in 20% GLY or in either concentration of EG (16–52%). In the toxicity test, exposure of tissues to DMSO, PROH or GLY resulted in higher percentages of normal follicles (80–97%) than exposure to EG (49%). Electron microscopy revealed damage to the ultrastructure of follicles frozen in 10% GLY, while follicles cryopreserved in DMSO and PROH at either concentration exhibited normal ultrastructure. In conclusion, DMSO and PROH were the most effective cryoprotectants for zebu ovarian tissue, preserving the structural integrity of somatic and reproductive cells within the ovary.