Helium ion microscopy visualizes lipid nanodomains in mammalian cells

Cell membranes are composed of two-dimensional bilayers of amphipathic lipids, which allow a lateral movement of the respective membrane components. These components are arranged in an inhomogeneous manner as transient micro- and nanodomains, which are believed to be crucially involved in the regulation of signal transduction pathways in mammalian cells. Because of their small size (diameter 10-200 nm), membrane nanodomains cannot be directly imaged using conventional light microscopy. Here, we present direct visualization of cell membrane nanodomains by helium ion microscopy (HIM). We show that HIM is capable to image biological specimens without any conductive coating, and that HIM images clearly allow the identification of nanodomains in the ultrastructure of membranes with 1.5 nm resolution. The shape of these nanodomains is preserved by fixation of the surrounding unsaturated fatty acids while saturated fatty acids inside the nanodomains are selectively removed. Atomic force microscopy, fluorescence microscopy, 3D structured illumination microscopy and direct stochastic optical reconstruction microscopy provide additional evidence that the structures in the HIM images of cell membranes originate from membrane nanodomains. The nanodomains observed by HIM have an average diameter of 20 nm and are densely arranged with a minimal nearest neighbor distance of ~15 nm.

Neotypifications of Linnaean names in Hybanthus (Violaceae)

Current studies in South American Violaceae detected the necessity of neotypes for Viola calceolaria L. (equivalent to Hybanthus calceolaria (L.) Oken) and Viola oppositifolia L. (equivalent to Hvbanthus oppositifolius (L.) Taub.), names of species originally described in Loefling`s her Hispanicum, but without validly published names.

Melatonin Protects CD4(+) T Cells from Activation-Induced Cell Death by Blocking NFAT-Mediated CD95 Ligand Upregulation

Over the past 20 y, the hormone melatonin was found to be produced in extrapineal sites, including cells of the immune system. Despite the increasing data regarding the biological effects of melatonin on the regulation of the immune system, the effect of this molecule on T cell survival remains largely unknown. Activation-induced cell death plays a critical role in the maintenance of the homeostasis of the immune system by eliminating self-reactive or chronically stimulated T cells. Because activated T cells not only synthesize melatonin but also respond to it, we investigated whether melatonin could modulate activation-induced cell death. We found that melatonin protects human and murine CD4(+) T cells from apoptosis by inhibiting CD95 ligand mRNA and protein upregulation in response to TCR/CD3 stimulation. This inhibition is a result of the interference with calmodulin/calcineurin activation of NFAT that prevents the translocation of NFAT to the nucleus. Accordingly, melatonin has no effect on T cells transfected with a constitutively active form of NFAT capable of migrating to the nucleus and transactivating target genes in the absence of calcineurin activity. Our results revealed a novel biochemical pathway that regulates the expression of CD95 ligand and potentially other downstream targets of NFAT activation. The Journal of Immunology, 2010, 184: 3487-3494.

Phylogeographical, ecological and biological patterns shown by nuclear (ssrRNA and gGAPDH) and mitochondrial (Cyt b) genes of trypanosomes of the subgenus Schizotrypanum parasitic in Brazilian bats

The genetic diversity and phylogeographical patterns of Trypanosoma species that infect Brazilian bats were evaluated by examining 1043 bats from 63 species of seven families captured in Amazonia, the Pantanal, Cerrado and the Atlantic Forest biomes of Brazil. The prevalence of trypanosonne-infected bats, as estimated by haemoculture, was 12.9%, resulting in 77 Cultures of isolates, most morphologically identified as Trypanosoma cf. cruzi, classified by barcoding using partial sequences from ssrRNA gene into the subgenus Schizotrypanum and identified as T. cruzi (15), T cruzi marinkellei (37) or T. cf. dionisii (25). Phylogenetic analyses using nuclear ssrRNA, glycosomal glyceraldehyde 3-phosphate dehydrogenase (gGAPDH) and mitochondrial cytochrome b (Cyt b) gene sequences generated three clades, which clustered together forming the subgenus Schizotrypanum. In addition to vector association, bat trypanosomes were related by the evolutionary history, ecology and phylogeography of the bats. Tryponosoma cf. dionisii trypanosomes (32.4%) infected 12 species from four bat families captured in all biomes, from North to South Brazil, and clustered with T. dionisii from Europe despite being separated by some genetic distance. Trypanosoma cruzi marinkellei (49.3%) was restricted to phyllostomid bats from Amazonia to the Pantanal (North to Central). Trypanosoma cruzi (18.2%) was found mainly in vespertilionid and phyllostomid bats from the Pantanal/Cerrado and the Atlantic Forest (Central to Southeast), with a few isolates from Amazonia. (C) 2009 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Phylogenetic Analyses Based on Small Subunit rRNA and Glycosomal Glyceraldehyde-3-Phosphate Dehydrogenase Genes and Ultrastructural Characterization of Two Snake Trypanosomes: Trypanosoma serpentis n. sp from Pseudoboa nigra and Trypanosoma cascavelli from Crotalus durissus terrificus

We sequenced the small subunit (SSU) rRNA and glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) genes of two trypanosomes isolated from the Brazilian snakes Pseudoboa nigra and Crotalus durissus terrificus. Trypanosomes were cultured and their morphometrical and ultrastructural features were characterized by light microscopy and scanning and transmission electron microscopy. Phylogenetic trees inferred using independent or combined SSU rRNA and gGAPDH data sets always clustered the snake trypanosomes together in a clade closest to lizard trypanosomes, forming a strongly supported monophyletic assemblage (i.e. lizard-snake clade). The positioning in the phylogenetic trees and the barcoding based on the variable V7-V8 region of the SSU rRNA, which showed high sequence divergences, allowed us to classify the isolates from distinct snake species as separate species. The isolate from P. nigra is described as a new species, Trypanosoma serpentis n. sp., whereas the isolate from C. d. terrificus is redescribed here as Trypanosoma cascavelli.

Phylogenetic analysis of Trypanosoma vivax supports the separation of South American/West African from East African isolates and a new T-vivax-like genotype infecting a nyala antelope from Mozambique

In this study, we addressed the phylogenetic and taxonomic relationships of Trypanosoma vivax and related trypanosomes nested in the subgenus Duttonella through combined morphological and phylogeographical analyses. We previously demonstrated that the clade T. vivax harbours a homogeneous clade comprising West African/South American isolates and the heterogeneous East African isolates. Herein we characterized a trypanosome isolated from a nyala antelope (Tragelaphus angasi) wild-caught in Mozambique (East Africa) and diagnosed as T. vivax-like based on biological, morphological and molecular data. Phylogenetic relationships, phylogeographical patterns and estimates of genetic divergence were based on SSU and ITS rDNA sequences of T. vivax from Brazil and Venezuela (South America), Nigeria (West Africa), and from T. vivax-like trypanosomes from Mozambique, Kenya and Tanzania (East Africa). Despite being well-supported within the T. vivax clade, the nyala trypanosome was highly divergent from all other T. vivax and T. vivax-like trypanosomes, even those from East Africa. Considering its host origin, morphological features, behaviour in experimentally infected goats, phylogenetic placement, and genetic divergence this isolate represents a new genotype of trypanosome closely phylogenetically related to T. vivax. This study corroborated the high complexity and the existence of distinct genotypes yet undescribed within the subgenus Duttonella.

Evolutionary history of trypanosomes from South American caiman (Caiman yacare) and African crocodiles inferred by phylogenetic analyses using SSU rDNA and gGAPDH genes

In this study, using a combined data set of SSU rDNA and gGAPDH gene sequences, we provide phylogenetic evidence that supports Clustering of crocodilian trypanosomes from the Brazilian Caiman yacare (Alligatoridae) and Trypanosoma grayi, a species that Circulates between African crocodiles (Crocodilydae) and tsetse flies. In a survey of trypanosomes in Caiman yacare from the Brazilian Pantanal, the prevalence of trypanosome infection was 35% as determined by microhaematocrit and haemoculture, and 9 cultures were obtained. The morphology of trypomastigotes from caiman blood and tissue imprints was compared with those described for other crocodilian trypanosomes. Differences in morphology and growth behaviour of caiman trypanosomes were corroborated by molecular polymorphism that revealed 2 genotypes. Eight isolates were ascribed to genotype Cay01 and 1 to genotype Cay02. Phylogenetic inferences based on concatenated SSU rDNA and gGAPDII sequences showed that caiman isolates are closely related to T. grayi, constituting a well-supported monophyletic assemblage (clade T. grayi). Divergence time estimates based on clade composition, and biogeographical and geological events were used to discuss the relationships between the evolutionary histories of crocodilian trypanosomes and their hosts.

Phylogeny of snake trypanosomes inferred by SSU rDNA sequences, their possible transmission by phlebotomines, and taxonomic appraisal by molecular, cross-infection and morphological analysis

Blood examination by microhaematocrit and haemoculture of 459 snakes belonging to 37 species revealed 24% trypanosome prevalence in species of Viperidae (Crotalus durissus and Bothrops jararaca) and Colubridae (Pseudoboa nigra). Trypanosome cultures from C. durissus and P. nigra were behaviourally and morphologically indistinguishable. In addition, the growth and morphological features of a trypanosome from the sand fly Viannaniyia tuberculata were similar to those of snake isolates. Cross-infection experiments revealed a lack of host restriction, as snakes of 3 species were infected with the trypanosome from C. durissus. Phylogeny based on ribosomal sequences revealed that snake trypanosomes clustered together with the sand fly trypanosome, forming a new phylogenetic lineage within Trypanosoma closest to a clade of lizard trypanosomes transmitted by sand flies dagger. The clade of trypanosomes from snakes and lizards suggests an association between the evolutionary histories of these trypanosomes and their squamate hosts. Moreover, data strongly indicated that these trypanosomes are transmitted by sand flies. The flaws of the current taxonomy of snake trypanosomes are discussed, and the need for molecular parameters to be adopted is emphasized. To our knowledge, this is the first molecular phylogenetic study of snake trypanosomes.

Study of the gamma radiation response of watch glasses

Some dosimetric properties of watch glasses were studied applying the thermoluminescence technique. The watch glass samples were powdered, and the selected grains were mixed with Teflon (TM). The mixture was pressed and sintered to produce pellets of watch glass-Teflon (TM) composites. The glow curves of the pellets show two peaks at 130 and 195 degrees C. Reproducibility of TL response was estimated to have a maximum coefficient of variation of 4.0%. The dose-response curve is sublinear between 0.5 and 20.0kGy. The calibration curve is linear between 1.0Gy and 1.0kGy. The minimum detection limits were also determined. The gamma radiation dose response and the thermal stability of the materials were studied with the purpose to establish the best conditions of watch glasses for use in gamma radiation dosimetry. (C) 2007 Elsevier Ltd. All rights reserved.

Influence of a prudent diet on circulating cathepsin S in humans

BACKGROUND: Increased circulating cathepsin S levels have been linked to increased risk of cardiometabolic diseases and cancer. However, whether cathepsin S is a modifiable risk factor is unclear. We aimed to investigate the effects of a prudent diet on plasma cathepsin S levels in healthy individuals. FINDINGS: Explorative analyses of a randomized study were performed in 88 normal to slightly overweight and hyperlipidemic men and women (aged 25 to 65) that were randomly assigned to ad libitum prudent diet, i.e. healthy Nordic diet (ND) or a control group (habitual Western diet) for 6 weeks. Whereas all foods in the ND were provided, the control group was advised to consume their habitual diet throughout the study. The ND was in line with dietary recommendations, e.g. low in saturated fats, sugars and salt, but high in plant-based foods rich in fibre and unsaturated fats.The ND significantly decreased cathepsin S levels (from 20.1 (+/-4.0 SD) to 19.7 μg/L (+/-4.3 SD)) compared with control group (from 18.2 (+/-2.9 SD) to 19.1 μg/L (+/-3.8 SD)). This difference remained after adjusting for sex and change in insulin sensitivity (P = 0.03), and near significant after adjusting for baseline cathepsin S levels (P = 0.06), but not for change in weight or LDL-C. Changes in cathepsin S levels were directly correlated with change in LDL-C. CONCLUSIONS: Compared with a habitual control diet, a provided ad libitum healthy Nordic diet decreased cathepsin S levels in healthy individuals, possibly mediated by weight loss or lowered LDL-C. These differences between groups in cathepsin S were however not robust and therefore need further investigation.

O dano moral nas relações de trabalho

O presente trabalho consiste em conceituar, demonstrar as características, objetividade do Dano e sua indenização. Por tratar-se de uma relação onde um lado é hipossuficiente, destaca-se, em face do presente estudo, a prática do ato ilícito pelo empregador. O Dano Moral, no Direito do Trabalho nasceu juntamente com a revolução industrial, onde constatamos mais efetivamente o aumento da subordinação e inicio das relações entre empregador e empregado, pois a partir daí surgem ambientes propícios para causar prejuízo a um bem extrapatrimonial, uma vez que tais relações se dão entre seres humanos capazes de ferir e de serem feridos na sua esfera intima, subjetiva. O estudo do presente tema, nos demonstra que os sujeitos da relação de trabalho são diferenciados um, o empregador, detentor do poder de comando, disciplina e fiscalização e o outro, o empregado, sujeito à subordinação, à disciplina e fiscalização. É justamente nesta diferenciação entre os sujeitos de tais relações que reside a origem do Dano Moral, pois, o mais fraco, o empregado, acaba sendo, pela prática do ato ilícito, prejudicado numa relação em que não respeita-se os limites entre as partes. Sendo o Dano Moral o resultado de uma ação ou omissão não estribada em exercício regular de um direito, em que o agente causa prejuízo ou viola direito de outrem, tem o autor da conduta lesionante a obrigação de repará-lo seja qual for a modalidade o dano. Assim, tanto os danos patrimoniais quanto os danos morais estão abrangidos no comando do artigo 927 do Código Civil.

São Pedro do Itabapoana: memória e identidade sul capixaba

Este trabalho tem como objeto de estudo São Pedro do Itabapoana, distrito pertencente ao município de Mimoso do Sul, localizado ao sul do estado do Espírito Santo. O objetivo da pesquisa é analisar três momentos importantes da sua história: a Revolução de 1930, o processo de tombamento do distrito como Sítio Histórico ocorrido em 1987 e o Festival de Inverno de Sanfona e Viola, criado em 1998. Através de pesquisa bibliográfica, jornais de época e, sobretudo, entrevistas de História Oral, foi possível explorar a relação entre patrimônio, memória e identidade. Ao mesmo tempo consegui identificar e reunir um acervo com informações pertinentes à região, que poderá contribuir para futuras pesquisas.

Práticas inovadoras utilizando a informática como expediente de otimização e modernização do poder judiciário: a utilização da videoconferência

Esta dissertação insere-se na linha de pesquisa Justiça e Sociedade do Programa de Pós-Graduação da Fundação Getúlio Vargas, Escola de Direito do Rio de Janeiro, Mestrado Profissional em Poder Judiciário, Área multidisciplinar. Teve como objeto de estudo o uso da videoconferência, analisando-a como prática inovadora de informática como expediente de otimização e modernização do Poder Judiciário. Quanto a metodologia empregou-se a pesquisa bibliográfica e de campo analisando medidas já implantadas em alguns Tribunais que possam ser utilizadas e aprimoradas em âmbito nacional. Examinou-se a controvérsia existente sobre a aplicação da videoconferência e conseqüências jurídicas e sociais do uso desta tecnologia. Com relação aos resultados concluiu-se que a utilização deste instrumento não viola princípios processuais, bem como não se trata de prática inconstitucional, mas sim gera benefícios para os cofres públicos e para o jurisdicionado, acarretando melhoria de qualidade na prestação jurisdicional

Improvisação teatral e descentração

Esta pesquisa tem por objetivo refletir sobre os processos cognitivos que envolvem a construção da improvisação teatral a partir do trabalho do ator, com base na teoria de Piaget e nos trabalhos de Viola Spolin. Como forma de compreender os processos cognitivos na construção da improvisação teatral, essa foi estudada a partir dos conceitos de egocentrismo e descentração propostos por Piaget. Através desses conceitos é possível traçar uma trajetória da construção da improvisação a partir do trabalho do ator. Esse estudo também se propõe a entender como ocorre a improvisação no que se refere às relações entre os atores e como esses constróem o jogo com os demais colegas. Para esta pesquisa, foi realizada uma oficina teatral como meio de recolher e registrar os dados necessários para as análises. Além da oficina teatral, foram feitas entrevistas semi-estruturadas e coletados dados através de cadernos de notas dos atores, o que possibilitou uma avaliação a partir da perspectiva, não só da pesquisadora, mas dos sujeitos envolvidos. As análises foram feitas a partir do trabalho empírico com base no quadro teórico escolhido. Dessa forma, foi possível identificar o percurso de construção do conhecimento em improvisação teatral, que se inicia com o jogo egocêntrico e evolui para o jogo cada vez mais descentrado e cooperativo.

Traduzindo Moll Flanders : as versões de Lucio Cardoso e Antônio Alves Cury sob a perspectiva dos estudos de tradução

Os fenômenos da tradução, assim como vários dos diversos percursos históricos e teóricos apresentados ao longo da História sobre o tema, serão explicitados e discutidos. A avaliação comparatista de duas traduções de Moll Flanders, de autoria de Daniel Defoe, será desenvolvida, levando-se em consideração as concepções mais atuais dos Estudos de Tradução, como a teoria da Reescritura de Andre Lefevere. Aspectos como o status do tradutor, assim como o sistema literário do qual este provém, são fatores que também serão pertinentes às referidas análises críticas.