Unique features of odorant-binding proteins of the parasitoid wasp Nasonia vitripennis revealed by genome annotation and comparative analyses

Insects are the most diverse group of animals on the planet, comprising over 90% of all metazoan life forms, and have adapted to a wide diversity of ecosystems in nearly all environments. They have evolved highly sensitive chemical senses that are central to their interaction with their environment and to communication between individuals. Understanding the molecular bases of insect olfaction is therefore of great importance from both a basic and applied perspective. Odorant binding proteins (OBPs) are some of most abundant proteins found in insect olfactory organs, where they are the first component of the olfactory transduction cascade, carrying odorant molecules to the olfactory receptors. We carried out a search for OBPs in the genome of the parasitoid wasp Nasonia vitripennis and identified 90 sequences encoding putative OBPs. This is the largest OBP family so far reported in insects. We report unique features of the N. vitripennis OBPs, including the presence and evolutionary origin of a new subfamily of double-domain OBPs (consisting of two concatenated OBP domains), the loss of conserved cysteine residues and the expression of pseudogenes. This study also demonstrates the extremely dynamic evolution of the insect OBP family: (i) the number of different OBPs can vary greatly between species; (ii) the sequences are highly diverse, sometimes as a result of positive selection pressure with even the canonical cysteines being lost; (iii) new lineage specific domain arrangements can arise, such as the double domain OBP subfamily of wasps and mosquitoes.

Cloning of human acetyl-CoA carboxylase-beta and its unique features.

Acetyl-CoA carboxylase, which has a molecular mass of 265 kDa (ACC-alpha), catalyzes the rate-limiting step in the biosynthesis of long-chain fatty acids. In this study we report the complete amino acid sequence and unique features of an isoform of ACC with a molecular mass of 275 kDa (ACC-beta), which is primarily expressed in heart and skeletal muscles. In these tissues, ACC-beta may be involved in the regulation of fatty acid oxidation, rather than fatty acid biosynthesis. ACC-beta contains an amino acid sequence at the N terminus which is about 200 amino acids long and may be uniquely related to the role of ACC-beta in controlling carnitine palmitoyltransferase I activity and fatty acid oxidation by mitochondria. If we exclude this unique sequence at the N terminus the two forms of ACC show about 75% amino acid identity. All of the known functional domains of ACC are found in the homologous regions. Human ACC-beta cDNA has an open reading frame of 7,343 bases, encoding a protein of 2,458 amino acids, with a calculated molecular mass of 276,638 Da. The mRNA size of human ACC-beta is approximately 10 kb and is primarily expressed in heart and skeletal muscle tissues, whereas ACC-alpha mRNA is detected in all tissues tested. A fragment of ACC-beta cDNA was expressed in Escherichia coli and antibodies against the peptide were generated to establish that the cDNA sequence that we cloned is that for ACC-beta.

The uncharged surface features surrounding the active site of Escherichia coli DsbA are conserved and are implicated in peptide binding

DsbA is a protein-folding catalyst from the periplasm of Escherichia coli that interacts with newly translocated polypeptide substrate and catalyzes the formation of disulfide bonds in these secreted proteins. The precise nature of the interaction between DsbA and unfolded substrate is not known. Here, we give a detailed analysis of the DsbA crystal structure, now refined to 1.7 Angstrom, and present a proposal for its interaction with peptide. The crystal structure of DsbA implies flexibility between the thioredoxin and helical domains that may be an important feature for the disulfide transfer reaction. A hinge point for domain motion is identified-the typo IV beta-turn Phe 63-Met 64-Gly 65-Gly 66, which connects the two domains. Three unique features on the active site surface of the DsbA molecule-a groove, hydrophobic pocket, and hydrophobic patch-form an extensive uncharged surface surrounding the active-sits disulfide. Residues that contribute to these surface features are shown to be generally conserved in eight DsbA homologues. Furthermore, the residues immediately surrounding the active-site disulfide are uncharged in all nine DsbA proteins. A model for DsbA-peptide interaction has been derived from the structure of a human thioredoxin:peptide complex. This shows that peptide could interact with DsbA in a manner similar to that with thioredoxin. The active-site disulfide and all three surrounding uncharged surface features of DsbA could, in principle, participate in the binding or stabilization of peptide.

Characterization of a unique variant of bat rabies virus responsible for newly emerging human cases in North America.

The silver-haired bat variant of rabies virus (SHBRV) has been identified as the etiological agent of a number of recent human rabies cases in the United States that are unusual in not having been associated with any known history of conventional exposure. Comparison of the different biological and biochemical properties of isolates of this virus with those of a coyote street rabies virus (COSRV) revealed that there are unique features associated with SHBRV. In vitro studies showed that, while the susceptibility of neuroblastoma cells to infection by both viruses was similar, the infectivity of SHBRV was much higher than that of COSRV in fibroblasts (BHK-21) and epithelial cells (MA-104), particularly when these cells were kept at 34 degrees C. At this temperature, low pH-dependent fusion and cell-to-cell spread of virus is seen in BHK-21 cells infected with SHBRV but not with COSRV. It appears that SHBRV may possess an unique cellular tropism and the ability to replicate at lower temperature, allowing a more effective local replication in the dermis. This hypothesis is supported by in vivo results which showed that while SHBRV is less neurovirulent than COSRV when administered via the intramuscular or intranasal routes, both viruses are equally neuroinvasive if injected intracranially or intradermally. Consistent with the above findings, the amino acid sequences of the glycoproteins of SHBRV and COSRV were found to have substantial differences, particularly in the region that contains the putative toxic loop, which are reflected in marked differences in their antigenic composition. Nevertheless, an experimental rabies vaccine based on the Pittman Moore vaccine strain protected mice equally well from lethal doses of SHBRV and COSRV, suggesting that currently used vaccines should be effective in the postexposure prophylaxis of rabies due to SHBRV.

An unusual, dwarf new species of Neotropical freshwater stingray, Plesiotrygon nana sp. nov., from the upper and mid Amazon basin: the second species of Plesiotrygon (Chondrichthyes: Potamotrygonidae)

A new species of the relatively poorly known Neotropical freshwater stingray genus Plesiotrygon Rosa, Castello & Thorson, 1987 is described from the main channel and smaller tributaries (Ríos Itaya and Pachitea) of the upper Amazon basin in Peru. The first specimen to be collected, however, was from much farther east in Rio Solimões in 1996, just down-river from Rio Purus (specimen unavailable for this study). Plesiotrygon nana sp. nov., is a very distinctive and unusually small species of freshwater stingray (Potamotrygonidae), described here mostly from three specimens representing different size classes and stages of sexual maturity. Plesiotrygon nana sp. nov., is distinguished from its only congener, P. iwamae Rosa, Castello & Thorson, 1987, by numerous unique features, including: dorsal coloration composed of very fine rosettes or a combination of spots and irregular ocelli; very circular disc and snout; very small and less rhomboidal spiracles; short snout and anterior disc region; narrow mouth and nostrils; denticles on dorsal tail small, scattered, not forming row of enlarged spines; adult and preadult specimens with significantly fewer tooth rows; fewer caudal vertebrae; higher total pectoral radials; very small size, probably not surpassing 250 mm disc length or width, males maturing sexually at around 180 mm disc length and 175 mm disc width; distal coloration of tail posterior to caudal stings usually dark purplish-brown; and features of the ventral lateral-line canals (hyomandibular canal very narrow, infraorbital and supraorbital canals not undulated, supraorbital and infraorbital loops small and narrow, supraorbital loop very short, not extending posteriorly to level of mouth, jugular and posterior infraorbital canals short, not extending caudally to first gill slits, subpleural loop very narrow posteriorly; absence of anterior and posterior subpleural tubules). To provide a foundation for the description of P. nana sp. nov., morphological variation in P. iwamae was examined based on all type specimens as well as newly collected and previously unreported material. Two specimens topotypic with the male paratype of P. nana sp. nov., referred to here as Plesiotrygon cf. iwamae, are also reported. Relationships of the new species to P. iwamae are discussed; further characters indicative of Plesiotrygon monophyly are proposed, but the genus may still not be valid. Plesiotrygon nana sp. nov., is commercialized with some regularity in the international aquarium trade from Iquitos (Peru), an alarming circumstance because nothing is known of its biology or conservation requirements.

Response of a two-level atom to a narrow-bandwidth squeezed-vacuum excitation

Using the coupled-system approach we calculate the optical spectra of the fluorescence and transmitted fields of a two-level atom driven by a squeezed vacuum of bandwidths smaller than the natural atomic linewidth. We find that in this regime of squeezing bandwidths the spectra exhibit unique features, such as a hole burning and a three-peak structure, which do not appear for a broadband excitation. We show that the features are unique to the quantum nature of the driving squeezed vacuum field and donor appear when the atom is driven by a classically squeezed field. We find that a quantum squeezed-vacuum field produces squeezing in the emitted fluorescence field which appears only in the squeezing spectrum while there is no squeezing in the total field. We also discuss a nonresonant excitation and find that depending on the squeezing bandwidth there is a peak or a hole in the spectrum at a frequency corresponding to a three-wave-mixing process. The hole appears only for a broadband excitation and results from the strong correlations between squeezed-vacuum photons.

Targeting the Acute Myeloid Leukemia Stem Cells

The idea that within the bulk of leukemic cells there are immature progenitors which are intrinsically resistant to chemotherapy and able to repopulate the tumor after treatment is not recent. Nevertheless, the term leukemia stem cells (LSCs) has been adopted recently to describe these immature progenitors based on the fact that they share the most relevant features of the normal hematopoetic stem cells (HSCs), i.e. the self-renewal potential and quiescent status. LSCs differ from their normal counterparts and from the more differentiated leukemic cells regarding the default status of pathways regulating apoptosis, cell cycle, telomere maintenance and transport pumps activity. In addition, unique features regarding the interaction of these cells with the microenvironment have been characterized. Therapeutic strategies targeting these unique features are at different stages of development but the reported results are promising. The aim of this review is, by taking acute myeloid leukemia (AML) as a bona fide example, to discuss some of the mechanisms used by the LSCs to survive and the strategies which could be used to eradicate these cells.

STANDARDIZATION OF SOME ELECTROCARDIOGRAPHIC PARAMETERS OF CAPTIVE LEOPARD CATS (LEOPARDUS TIGRINUS)

Thirty-three captive leopard cats, Leopardus tigrinus, were anesthetized with xylazine (1-2 mg/kg) and ketamine (10 mg/kg), and electrocardiograph (ECG) tests were recorded in all leads with 1 cm = 1 mV sensibility and 25 mm/sec speed repeating DII lead at 50 mm/sec speed with the same sensibility. Results expressed by mean and standard deviation were: heart rate (HR) = 107 +/- 17 (bpm); P-wave = 0.048 +/- 0.072 (s) x 0.128 +/- 0.048 (mV); PR interval = 0.101 +/- 0.081 (s); QRS compound = 0.053 +/- 0.012 (s) x 1.446 +/- 0.602 (mV); QT interval = 0.231 +/- 0.028 (s); R-wave (CV(6)LL) = 1.574 +/- 0.527 (mV); R-wave (CV(6)LU) = 1.583 +/- 0.818 (mV); heart rhythm: normal sinus rhythm (15.2%), sinus rhythm with wandering pacemaker (WPM) (60.6%), sinus arrhythmia with WPM (24.2%); electric axis: between +30 degrees and +60 degrees (6.1%), +60 (6.1%), between +60 degrees and +90 degrees (57.6%), +90 degrees (9%), between +90 degrees and +120 degrees (21.2%); ST segment: normal (75.7%), elevation (18.2%), depression (6.1%); T-wave polarity (DII): positive (100%); T-wave (V(10)): absent (6.1%), negative (63.6%), positive (18.2%), and with interference (12.1%). Through ECG data comparison with other species, unique features of Leopardus tigrinus` (leopard cat) ECG parameters were detected. Some of the study animals presented with an R-Wave amplitude that was indicative of left ventricle overload according to patterns for normal domestic cats (Felis cati). Echocardiographic exams revealed normal heart cavities` function and morphology. The aim of this study was to establish some electrocardiographic parameters of captive L. tigrinus.

Is Cadinella ornatissima a chomodorid? Sperm ultra structure in an enigmatic nudibranch (Opisthobranchia Mollusca)

Sperm ultrastructure is described for the nudibranch gastropod Cadlinella ornatissima, type species of the genus Cadlinella (Thiele). Although C. ornatissima exhibits most of the sperm features characteristic of other Opisthobranchia and the Pulmonata (a small, rounded acrosomal vesicle, a complex, helical, mitochondrial derivative - partially paracrystalline, coarse fibres associated with the axoneme), it also possesses a number of previously undescribed and possibly unique features (a longitudinally inrolled acrosomal pedestal, an axial structure within the cavity of the acrosomal pedestal, an electron-dense collar at the anterior region of the acrosomal pedestal, the presence of crystalloid bodies within the glycogen helices of the mitochondrial derivative). To our knowledge this is the first report of crystalloid bodies in mature sperm of any mollusc. Collectively this evidence raises questions concerning the affinities and systematic position of Cadlinella within the Nudibranchia. The peculiar nature of the sperm differences, in comparison with other investigated nudibranchs, suggest that Cadlinella is not easily linked to either the Cadlinidae or Chromodorididae, and should be considered incertae sedis.

Candidatus "Scalindua brodae", spec. nov., Candidatus "Scalindua wagneri", spec. nov., two new species of anaerobic ammonium oxidizing bacteria

Anaerobic ammonium oxidation (anammox) is both a promising process in wastewater treatment and a long overlooked microbial physiology that can contribute significantly to biological nitrogen cycling in the world's oceans. Anammox is mediated by a monophyletic group of bacteria that branches deeply in the Planctomycetales. Here we describe a new genus and species of anaerobic ammonium oxidizing planctomycetes, discovered in a wastewater treatment plant (wwtp) treating landfill leachate in Pitsea, UK. The biomass from this wwtp showed high anammox activity (5.0 +/- 0.5 nmol/mg protein/min) and produced hydrazine from hydroxylamine, one of the unique features of anammox bacteria. Eight new planctomycete 16S rRNA gene sequences were present in the 16S rRNA gene clone library generated from the biomass. Four of these were affiliated to known anammox 16S rRNA gene sequences, but branched much closer to the root of the planctomycete line of descent. Fluorescence in situ hybridization (FISH) with oligonucleotide probes specific for these new sequences showed that two species (belonging to the same genus) together made up > 99% of the planctomycete population which constituted 20% of the total microbial community. The identification of these organisms as typical anammox bacteria was confirmed with electron microscopy and lipid analysis. The new species, provisionally named Candidatus Scalindua brodae and Scalindua wagneri considerably extend the biodiversity of the anammox lineage on the 16S rRNA gene level, but otherwise resemble known anammox bacteria. Simultaneously, another new species of the same genus, Candidatus Scalindua sorokinii, was detected in the water column of the Black Sea, making this genus the most widespread of all anammox bacteria described so far.

Evas??o em cursos de educa????o continuada a dist??ncia: um estudo na Escola Nacional de Administra????o P??blica

A an??lise da literatura brasileira sobre educa????o a dist??ncia ?? ainda limitada no que diz respeito a estudos que investiguem o fen??meno da evas??o, em especial, em contextos de escolas de governo. Este artigo procura identificar e analisar os fatores que contribuem para a evas??o de participantes nos cursos de educa????o continuada a dist??ncia em uma organiza????o governamental respons??vel pela capacita????o dos servidores da administra????o p??blica federal. A ??nfase das investiga????es encontra-se nas vari??veis referentes ??s dificuldades pessoais, profissionais e tecnol??gicas, ??s peculiaridades do curso e caracter??sticas da clientela, contrastadas com as vari??veis demogr??ficas referentes ?? evas??o em cursos a dist??ncia mediados por tecnologias da informa????o e da comunica????o. O trabalho ?? embasado em metodologia quantitativa, por meio de pesquisa descritiva de campo junto aos participantes, valendo-se do uso de question??rio eletr??nico. Procura contribuir para investiga????es sobre o tema, em especial no que se refere aos servidores p??blicos.

Mob1: defining cell polarity for proper cell division

Mob1 is a component of both the mitotic exit network and Hippo pathway, being required for cytokinesis, control of cell proliferation and apoptosis. Cell division accuracy is crucial in maintaining cell ploidy and genomic stability and relies on the correct establishment of the cell division axis, which is under the control of the cell's environment and its intrinsic polarity. The ciliate Tetrahymena thermophila possesses a permanent anterior-posterior axis, left-right asymmetry and divides symmetrically. These unique features of Tetrahymena prompted us to investigate the role of Tetrahymena Mob1. Unexpectedly, we found that Mob1 accumulated in basal bodies at the posterior pole of the cell, and is the first molecular polarity marker so far described in Tetrahymena. In addition, Mob1 depletion caused the abnormal establishment of the cell division plane, providing clear evidence that Mob1 is important for its definition. Furthermore, cytokinesis was arrested and ciliogenesis delayed in Tetrahymena cells depleted of Mob1. This is the first evidence for an involvement of Mob1 in cilia biology. In conclusion, we show that Mob1 is an important cell polarity marker that is crucial for correct division plane placement, for cytokinesis completion and for normal cilia growth rates.

Optimized near real-time remote device tracking and monitoring system

Os serviços baseados em localização vieram dar um novo alento à criatividade dos programadores de aplicações móveis. A vulgarização de dispositivos com capacidades de localização integradas deu origem ao desenvolvimento de aplicações que gerem e apresentam informação baseada na posição do utilizador. Desde então, o mercado móvel tem assistido ao aparecimento de novas categorias de aplicações que tiram proveito desta capacidade. Entre elas, destaca-se a monitorização remota de dispositivos, que tem vindo a assumir uma importância crescente, tanto no sector particular como no sector empresarial. Esta dissertação começa por apresentar o estado da arte sobre os diferentes sistemas de posicionamento, categorizados pela sua eficácia em ambientes internos ou externos, assim como diferentes protocolos de comunicação em tempo quase-real. É também feita uma análise ao estado actual do mercado móvel. Actualmente o mercado possui diferentes plataformas móveis com características únicas que as fazem rivalizar entre si, com vista a expandirem a sua quota de mercado. É por isso elaborado um breve estudo sobre os sistemas operativos móveis mais relevantes da actualidade. É igualmente feita uma abordagem mais profunda à arquitectura da plataforma móvel da Apple - o iOS – que serviu de base ao desenvolvimento de uma solução optimizada para localização e monitorização de dispositivos móveis. A monitorização implica uma utilização intensiva de recursos energéticos e de largura de banda que os dispositivos móveis da actualidade não estão aptos a suportar. Dado o grande consumo energético do GPS face à precária autonomia destes dispositivos, é apresentado um estudo em que se expõem soluções que permitem gerir de forma optimizada a utilização do GPS. O elevado custo dos planos de dados facultados pelas operadoras móveis é também considerado, pelo que são exploradas soluções que visam minimizar a utilização de largura de banda. Deste trabalho, nasce a aplicação EyeGotcha, que para além de permitir localizar outros utilizadores de dispositivos móveis de forma optimizada, permite também monitorizar as suas acções baseando-se num conjunto de regras pré-definidas. Estas acções são reportadas às entidades monitoras, de modo automatizado e sob a forma de alertas. Visionando-se a comercialização da aplicação, é portanto apresentado um modelo de negócio que permite obter receitas capazes de cobrirem os custos de manutenção de serviços, aos quais o funcionamento da aplicação móvel está subjugado.

A passagem à reforma : um estudo exploratório sobre mulheres profissionalizadas na sociedade portuguesa

RESUMO: Devido às mudanças políticas e sociais que ocorreram no passado, a proporção de mulheres activas no mercado de trabalho tem vindo a aumentar, e neste sentido, cada vez mais mulheres têm vindo a entrar na reforma. As recentes evoluções demográficas mostram um crescente envelhecimento populacional caracterizado por um aumento da proporção de pessoas idosas e pela sua maior longevidade. As mulheres são em número superior, no entanto, a realidade da mulher portuguesa reformada tem sido pouco avaliada sob o ponto de vista em que decorre esta transição. A passagem à reforma é um momento fulcral para conhecer como se adaptam os indivíduos a uma nova etapa da sua vida que é actualmente vivida por mais tempo, e que representa também a passagem para outra categoria social, a categoria de reformado. Condicionantes sociais, culturais e individuais, contribuem para modelar esta transição e o ajustamento à mesma. A reforma para as mulheres deverá corresponder a uma etapa com características únicas, devido às particularidades em termos profissionais e sociais que as distinguem dos homens. Pretende-se neste trabalho “dar voz” às mulheres portuguesas que tiveram uma carreira profissional e conhecer as suas experiências de transição para a reforma e a forma como vivem esta condição. Foram realizadas entrevistas em profundidade com mulheres portuguesas profissionais reformadas, cujos conteúdos foram analisados em torno das seguintes categorias: sentimentos vividos; planeamento e motivações para a passagem à reforma; relação com o trabalho; noção de si própria; gestão de tempo e organização quotidiana e interacções familiares e sociais.-------- ABSTRACT: Due to past political and social changes the number of women working actively in the labor market is growing. This implies that, more women are also entering in the retirement period. Recent demographic trends show an increasing ageing population, characterized by a higher proportion of elderly people, and a higher longevity. Women’s proportion outnumbers older men, yet the reality of Portuguese retired women has been poorly evaluated in regard to this transition process. Retirement transition is a crucial period to understand how individuals adapt to a new stage in their life, that is actually being enjoyed for a longer period and that also represents the transition to retiree’s social role. Social, cultural and individual conditions help to shape this transition and adjustment to it. Retirement for women should be an event with unique features, mostly because of the peculiarities in professional and social relationships, distinct from men. Through in-depth interviews, we explored how Portuguese women, who had a professional career, experience the retirement transition and how they live this new condition. The women’s narratives were analyzed within the following categories: experienced feelings, planning and motivation for retirement; notion of self; time management and daily organization; family and social interactions.

Heavy-metal resistance in Marinobacter aquaeolei 617 insights into copper resistance

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Biotecnologia