Ecuador Paraiso Escondido virus, a new flavivirus isolated from New World sand flies in Ecuador, is the first representative of a novel clade in the genus flavivirus

A new flavivirus, Ecuador Paraiso Escondido virus (EPEV), named after the village where it was discovered, was isolated from sand flies (Psathyromyia abonnenci, formerly Lutzomyia abonnenci) that are unique to the New World. This represents the first sand fly-borne flavivirus identified in the New World. EPEV exhibited a typical flavivirus genome organization. Nevertheless, the maximum pairwise amino acid sequence identity with currently recognized flaviviruses was 52.8%. Phylogenetic analysis of the complete coding sequence showed that EPEV represents a distinct clade which diverged from a lineage that was ancestral to the nonvectored flaviviruses Entebbe bat virus, Yokose virus, and Sokoluk virus and also the Aedes-associated mosquito-borne flaviviruses, which include yellow fever virus, Sepik virus, Saboya virus, and others. EPEV replicated in C6/36 mosquito cells, yielding high infectious titers, but failed to reproduce either in vertebrate cell lines (Vero, BHK, SW13, and XTC cells) or in suckling mouse brains. This surprising result, which appears to eliminate an association with vertebrate hosts in the life cycle of EPEV, is discussed in the context of the evolutionary origins of EPEV in the New World.The flaviviruses are rarely (if ever) vectored by sand fly species, at least in the Old World. We have identified the first representative of a sand fly-associated flavivirus, Ecuador Paraiso Escondido virus (EPEV), in the New World. EPEV constitutes a novel clade according to current knowledge of the flaviviruses. Phylogenetic analysis of the virus genome showed that EPEV roots the Aedes-associated mosquito-borne flaviviruses, including yellow fever virus. In light of this new discovery, the New World origin of EPEV is discussed together with that of the other flaviviruses.

An assessment of the latent and sensible heat flux on the simulated regional climate over Southwestern South Atlantic Ocean

A Regional Climate Model (RegCM3) 10-year (1990-1999) simulation over southwestern South Atlantic Ocean (SAO) is evaluated to assess the mean climatology and the simulation errors of turbulent fluxes over the sea. Moreover, the relationship between these fluxes and the rainfall over some cyclogenetic areas is also analyzed. The RegCM3 results are validated using some reanalyses datasets (ERA40, R2, GPCP and WHOI). The summer and winter spatial patterns of latent and sensible heat fluxes simulated by the RegCM3 are in agreement with the reanalyses (WHOI, R2 and ERA40). They show large latent heat fluxes exchange in the subtropical SAO and at higher latitudes in the warm waters of Brazil Current. In particular, the magnitude of RegCM3 latent heat fluxes is similar to the WHOI, which is probably related to two factors: (a) small specific humidity bias, and (b) the RegCM3 flux algorithm. In contrast, the RegCM3 presents large overestimation of sensible heat flux, though it simulates well their spatial pattern. This simulation error is associated with the RegCM3 underestimation of the 2-m air temperature. In southwestern SAO, in three known cyclogenetic areas, the reanalyses and the RegCM3 show the existence of different physical mechanisms that control the annual cycles of latent/sensible heating and rainfall. It is shown that over the eastern coast of Uruguay (35A degrees-43A degrees S) and the southeastern coast of Argentina (44A degrees-52A degrees S) the sea-air moisture and heat exchange play an important role to control the annual cycle of precipitation. This does not happen on the south/southeastern coast of Brazil.

IPCC global coupled model simulations of the South America monsoon system

This study examines the variability of the South America monsoon system (SAMS) over tropical South America (SA). The onset, end, and total rainfall during the summer monsoon are investigated using precipitation pentad estimates from the global precipitation climatology project (GPCP) 1979-2006. Likewise, the variability of SAMS characteristics is examined in ten Intergovernmental Panel on Climate Change (IPCC) global coupled climate models in the twentieth century (1981-2000) and in a future scenario of global change (A1B) (2081-2100). It is shown that most IPCC models misrepresent the intertropical convergence zone and therefore do not capture the actual annual cycle of precipitation over the Amazon and northwest SA. Most models can correctly represent the spatiotemporal variability of the annual cycle of precipitation in central and eastern Brazil such as the correct phase of dry and wet seasons, onset dates, duration of rainy season and total accumulated precipitation during the summer monsoon for the twentieth century runs. Nevertheless, poor representation of the total monsoonal precipitation over the Amazon and northeast Brazil is observed in a large majority of the models. Overall, MI-ROC3.2-hires, MIROC3.2-medres and MRI-CGCM3.2.3 show the most realistic representation of SAMS`s characteristics such as onset, duration, total monsoonal precipitation, and its interannual variability. On the other hand, ECHAM5, GFDL-CM2.0 and GFDL-CM2.1 have the least realistic representation of the same characteristics. For the A1B scenario the most coherent feature observed in the IPCC models is a reduction in precipitation over central-eastern Brazil during the summer monsoon, comparatively with the present climate. The IPCC models do not indicate statistically significant changes in SAMS onset and demise dates for the same scenario.

Habitat Split as a Cause of Local Population Declines of Amphibians with Aquatic Larvae

Most amphibian species have biphasic life histories and undergo an ontogenetic shift from aquatic to terrestrial habitats. In deforested landscapes, streams and forest fragments are frequently disjunct, jeopardizing the life cycle of forest-associated amphibians with aquatic larvae. We tested the impact of habitat split-defined as human-induced disconnection between habitats used by different life-history stages of a species-on four forest-associated amphibian species in a severely fragmented landscape of the Brazilian Atlantic Forest. We surveyed amphibians in forest fragments with and without streams (referred to as wet and dry fragments, respectively), including the adjacent grass-field matrix. Our comparison of capture rates in dry fragments and nearby streams in the matrix allowed us to evaluate the number of individuals that engaged in high-risk migrations through nonforested habitats. Adult amphibians moved from dry fragments to matrix streams at the beginning of the rainy season, reproduced, and returned at the end of the breeding period. Juveniles of the year moved to dry fragments along with adults. These risky reproductive migrations through nonforested habitats that expose individuals to dehydration, predation, and other hazards may cause population declines in dry fragments. Indeed, capture rates were significantly lower in dry fragments compared with wet fragments. Declining amphibians would strongly benefit from investments in the conservation and restoration of riparian vegetation and corridors linking breeding and nonbreeding areas.

Domestic Cats Constitute a Natural Reservoir of Human Enteropathogenic Escherichia coli Types

Feces of 70 diarrhoeic and 230 non-diarrhoeic domestic cats from Sao Paulo, Brazil were investigated for enteropathogenic (EPEC), enterohaemorrhagic (EHEC) and enterotoxigenic (ETEC) Escherichia coli types. While ETEC and EHEC strains were not found, 15 EPEC strains were isolated from 14 cats, of which 13 were non-diarrhoeic, and one diarrhoeic. None of 15 EPEC strains carried the bfpA gene or the EPEC adherence factor plasmid, indicating atypical EPEC types. The EPEC strains were heterogeneous with regard to intimin types, such as eae-theta (three strains), eae-kappa (n = 3), eae-alpha 1 (n = 2), eae-iota (n = 2), one eae-alpha 2, eae-beta 1 and eae-eta each, and two were not typeable. The majority of the EPEC isolates adhered to HEp-2 cells in a localized adherence-like pattern and were positive for fluorescence actin staining. The EPEC strains belonged to 12 different serotypes, including O111:H25 and O125:H6, which are known to be pathogens in humans. Multi locus sequence typing revealed a close genetic similarity between the O111:H25 and O125:H6 strains from cats, dogs and humans. Our results show that domestic cats are colonized by EPEC, including serotypes previously described as human pathogens. As these EPEC strains are also isolated from humans, a cycle of mutual infection by EPEC between cats and its households cannot be ruled out, though the transmission dynamics among the reservoirs are not yet understood clearly.

Rural Triatoma rubrovaria from Southern Brazil harbors Trypanosoma cruzi of lineage IIc

Triatoma infestans, the main vector of Chagas disease, has nearly been eliminated from Brazil. Nevertheless, other triatominae species are involved in the domiciliation process, including Triatoma rubrovaria in Rio Grande do Sul State (RS). Previous studies showed that 1.6% of the T rubrovaria specimens collected at the rural district of Quarai, RS, were naturally infected by Trypanosoma cruzi. In this study, five T. cruzi isolates obtained from infected triatomines were characterized molecularly and biologically. Genotyping of the T cruzi isolates showed that they belong to lineage IIc of T cruzi (TCIIc). Biological characterization showed miotropism and myositis during acute and chronic phases of infection, respectively. Virulence and mortality rates were variable among isolates. To our knowledge, this study corresponds to the first characterization of T cruzi isolates from T rubrovaria and the first description of TCIIc in the sylvatic cycle of T cruzi from the southern region of Brazil.

The Estrous Cycle Modulates Small Leucine-Rich Proteoglycans Expression in Mouse Uterine Tissues

In the pregnant mouse uterus, small leucine-rich proteoglycans (SLRPs) are drastically remodeled within a few hours after fertilization, suggesting that ovarian hormone levels modulate their synthesis and degradation. In this study, we followed by immunoperoxidase approach, the presence of four members of the SLRP family (decorin, lumican, biglycan, and fibromodulin) in the uterine tissues along the estrous cycle of the mouse. All molecules except fibromodulin, which predominates in the myometrium, showed a striking modulation in their distribution in the endometrial stroma, following the rise in the level of estrogen. Moreover, notable differences in the distribution of SLRPs were observed between superficial and deep stroma, as well as between the internal and external layers of the myometrium. Only biglycan and fibromodulin were expressed in the luminal and glandular epithelia. All four SLRPs were found in cytoplasmic granules of mononucleated cells. The pattern of distribution of the immunoreaction for these molecules in the uterine tissues was found to be estrous cycle-stage dependent, suggesting that these molecules undergo ovarian hormonal control and probably participate in the preparation of the uterus for decidualization and embryo implantation. In addition, this and previous results from our laboratory suggest the existence of two subpopulations of endometrial fibroblasts that may be related to the centrifugal development of the decidua. Anat Rec, 292:138-153, 2009. (c) 2008 Wiley-Liss, Inc.

Proteins and Peptidases from Conidia and Mycelia of Scedosporium apiospermum Strain HLPB

The conidia-mycelia transformation is an essential step during the life cycle of the fungal human pathogens of the Pseudallescheria boydii complex. In the present study, we have analyzed the protein and peptidase profiles in two distinct morphological stages, conidia and mycelia, of Scedosporium apiospermum sensu stricto. Proteins synthesized by the mycelia, migrating at the ranges of 62-48 and 22-18 kDa, were not detected from the conidial extract. Conidia produced a single cellular peptidase of 28 kDa able to digest copolymerized albumin, while mycelia yielded 6 distinct peptidases ranging from 90 to 28 kDa. All proteolytic enzymes were active at acidic pH and fully inhibited by 1,10-phenanthroline, characterizing these activities as metallo-type peptidases. Quantitative peptidase assay, using soluble albumin, showed a high metallopeptidase production in mycelial cells in comparison with conidia. The regulated expression of proteins and peptidases in different morphological stages of S. apiospermum represents a potential target for isolation of stage-specific markers for biochemical and immunological analysis.

Discovery of novel Trypanosoma cruzi glyceraldehyde-3-phosphate dehydrogenase inhibitors

Based on its essential role in the life cycle of Trypanosoma cruzi, the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH) has been considered a promising target for the development of novel chemotherapeutic agents for the treatment of Chagas` disease. In the course of our research program to discover novel inhibitors of this trypanosomatid enzyme, we have explored a combination of structure and ligand-based virtual screening techniques as a complementary approach to a biochemical screening of natural products using a standard biochemical assay. Seven natural products, including anacardic acids,. avonoid derivatives, and one glucosylxanthone were identified as novel inhibitors of T. cruzi GAPDH. Promiscuous inhibition induced by nonspecific aggregation has been discarded as specific inhibition was not reversed or affected in all cases in the presence of Triton X-100, demonstrating the ability of the assay to find authentic inhibitors of the enzyme. The structural diversity of this series of promising natural products is of special interest in drug design, and should therefore be useful in future medicinal chemistry efforts aimed at the development of new GAPDH inhibitors having increased potency. (C) 2009 Elsevier Ltd. All rights reserved.

Anacardic acid derivatives as inhibitors of glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma cruzi

Chagas` disease, a parasitic infection caused by the flagellate protozoan Trypanosoma cruzi, is a major public health problem affecting millions of individuals in Latin America. On the basis of the essential role in the life cycle of T. cruzi, the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has been considered an attractive target for the development of novel antitrypanosomatid agents. In the present work, we describe the inhibitory effects of a small library of natural and synthetic anacardic acid derivatives against the target enzyme. The most potent inhibitors, 6-n-pentadecyl-(1) and 6-n-dodecylsalicilic acids (10e), have IC(50) values of 28 and 55 mu M, respectively. The inhibition was not reversed or prevented by the addition of Triton X-100, indicating that aggregate-based inhibition did not occur. In addition, detailed mechanistic characterization of the effects of these compounds on the T. cruzi GAPDH-catalyzed reaction showed clear noncompetitive inhibition with respect to both substrate and cofactor. (C) 2008 Elsevier Ltd. All rights reserved.

Glucose uptake in the mammalian stages of Trypanosoma cruzi

Trypanosoma cruzi, the agent of Chagas` disease, alternates between different morphogenetic stages that face distinct physiological conditions in their invertebrate and vertebrate hosts, likely in the availability of glucose. While the glucose transport is well characterized in epimastigotes of T cruzi, nothing is known about how the mammalian stages acquire this molecule. Herein glucose transport activity and expression were analyzed in the three developmental stages present in the vertebrate cycle of T cruzi. The infective trypomastigotes showed the highest transport activity (V(max) = 5.34 +/- 0.54 nmol/min per mg of protein: K(m) = 0.38 +/- 0.01 mM) when compared to intracellular epimastigotes (V(max) = 2.18 +/- 0.20 nmol/min per mg of protein; K(m) = 0.39 +/- 0.01 mM). Under the conditions employed no transport activity could be detected in amastigotes. The gene of the glucose transporter is expressed at the mRNA level in trypomastigotes and in intracellular epimastigotes but not in amastigotes, as revealed by real-time PCR. In both trypomastigotes and intracellular epimastigotes protein expression could be detected by Western blot with an antibody raised against the glucose transporter correlating well with the transport activity measured experimentally. Interestingly, anti-glucose transporter antibodies showed a strong reactivity with glycosome and reservosome organelles. A comparison between proline and glucose transport among the intracellular differentiation forms is presented. The data suggest that the regulation of glucose transporter reflects different energy and carbon requirements along the intracellular life cycle of T cruzi. (C) 2009 Elsevier B.V. All rights reserved.

Implementing stepwise solvent elution in sequential injection chromatography for fluorimetric determination of intracellular free amino acids in the microalgae Tetraselmis gracilis

The concept of sequential injection chromatography (SIC) was exploited to automate the fluorimetric determination of amino acids after pre-column derivatization with ophthaldialdehyde (OPA) in presence of 2-mercaptoethanol (2MCE) using a reverse phase monolithic C(18) stationary phase. The method is low-priced and based on five steps of isocratic elutions. The first step employs the mixture methanol: tetrahydrofuran: 10 mmol L(-1) phosphate buffer (pH 7.2) at the volumetric ratio of 8:1:91; the other steps use methanol: 10 mmol L-1 phosphate buffer (pH 7.2) at volumetric ratios of 20:80, 35:65, SO:SO and 65:35. At a flow rate of 10 mu L s(-1) a 25 mm long-column was able to separate aspartic acid (Asp), glutamic acid (Glu), asparagine (Asn), serine (Ser), glutamine (Gln), glycine (Gly), threonine (Thr), citruline (Ctr), arginine (Arg), alanine (Ala), tyrosine (Tyr), phenylalanine (Phe), ornithine (Orn) and lysine (Lys) with resolution >1.2 as well as methionine (Met) and valine (Val) with resolution of 0.6. Under these conditions isoleucine (Ile) and leucine (Leu) co-eluted. The entire cycle of amino acids derivatization, chromatographic separation and column conditioning at the end of separation lasted 25 min. At a flow rate of 40 mu L s(-1) such time was reduced to 10 min at the cost of resolution worsening for the pairs Ctr/Arg and Orn/Lys. The detection limits varied from 0.092 mu mol L(-1) for Tyr to 0.51 mu mol L(-1) for Orn. The method was successfully applied to the determination of intracellular free amino acids in the green alga Tetraselmis gracilis during a period of seven days of cultivation. Samples spiked with known amounts of amino acids resulted in recoveries between 94 and 112%. (C) 2008 Elsevier B.V. All rights reserved.

Natural Theology: A Reevaluation of Christianity's Relationship to the Natural World

In this thesis I offer two separate arguments for the creation of an environmentally friendly Christian theology. These arguments, although interconnected, are roughly divided into the main chapters of the thesis. I will begin in Chapter Two by offering a negative argument against the assumption that the natural world is sinful. In their article Hauerwas and Berkman suggest that the suffering of animals is both an example of the sinful state of the environment and a justification for human separation from an unholy natural environment. In response to this view I will argue in the second chapter that the suffering of animals can be seen as part of God's intentions for our world. Suffering, in both the human and the larger world, is not evidence of a fundamental flaw in natural systems. Instead, the cycle of death and life found in the natural world can be profoundly spiritual.

Development Of Climate Change Information Database And Its Use In Civic Consciousness Enlightenment

GCM outputs such as CMIP3 are available via network access to PCMDI web site. Meteorological researchers are familiar with the usage of the GCM data, but the most of researchers other than meteorology such as agriculture, civil engineering, etc., and general people are not familiar with the GCM. There are some difficulties to use GCM; 1) to download the enormous quantity of data, 2) to understand the GCM methodology, parameters and grids. In order to provide a quick access way to GCM, Climate Change Information Database has been developed. The purpose of the database is to bridge the users and meteorological specialists and to facilitate the understanding the climate changes. The resolution of the data is unified, and climate change amount or factors for each meteorological element are provided from the database. All data in the database are interpolated on the same 80km mesh. Available data are the present-future projections of 27 GCMs, 16 meteorological elements (precipitation, temperature, etc.), 3 emission scenarios (A1B, A2, B1). We showed the summary of this database to residents in Toyama prefecture and measured the effect of showing and grasped the image for the climate change by using the Internet questionary survey. The persons who feel a climate change at the present tend to feel the additional changes in the future. It is important to show the monitoring results of climate change for a citizen and promote the understanding for the climate change that had already occurred. It has been shown that general images for the climate change promote to understand the need of the mitigation, and that it is important to explain about the climate change that might occur in the future even if it did not occur at the present in order to have people recognize widely the need of the adaptation.

Cozinha da Madeira website: the online touchpoint for a sustainability oriented service of Madeira Island

Over the past several years technology has been evolving in a way that it has become crucial for most businesses and companies to have interactive technology enabled touchpoints available online. Such interactive touchpoints can be developed as mobile application, webpages, or even through social networks. In the end such touchpoints will most surely represent the most easily reachable and marketable side of the business. Today selling a product alone is no longer enough to make consumers satisfied and complete, businesses and business models are changing. Increasingly, companies are choosing to not just sell products but to combine both sale and service. These service-based approaches will provide the client with a unique and personalized experience of what the company is selling. By selling a service the company transmits values that are more complex than the simple selling of a product. A service is something immaterial, happens over time and exists in the moment of the delivery. When conceiving and designing services, the use of the new technologies becomes a crucial step in order to craft touchpoints that facilitate the whole experience cycle of the service, from attracting, orienting, interacting and retaining the client, as well as providing later support to the consumer to advocate for the service itself. This thesis reports on the design and implementation of the online touchpoint of Cozinha da Madeira, which is a service designed to support tourism, specifically promoting the discovery of tradition and landscapes in the island of Madeira. Such touchpoint developed in the form of a website, embodies completely or partially various stages of the Service Experience cycle, from attracting and connecting, orienting, interacting as well as retaining and advocating. Through this thesis we will describe the design and implementation of such touchpoint as well as the evaluation and possible future implications.