Biochemical and biophysical properties of a highly active recombinant arginase from Leishmania (Leishmania) amazonensis and subcellular localization of native enzyme

Arginase (L-arginine amidinohydrolase, E.C. 3.5.3.1) is a metalloenzyme that catalyses the hydrolysis Of L-arginine to L-ornithine and urea. In Leishmania spp., the biological role of the enzyme may be involved in modulating NO production upon macrophage infection. Previously, we cloned and characterized the arginase gene from Leishmania (Leishmania) amazonensis. In the present work, we successfully expressed the recombinant enzyme in E. coli and performed biochemical and biophysical characterization of both the native and recombinant enzymes. We obtained K-M and V-max. values of 23.9(+/- 0.96) mM and 192.3 mu mol/min mg protein (+/- 14.3), respectively, for the native enzyme. For the recombinant counterpart, K-M was 21.5(+/- 0.90) mM and V-max was 144.9(+/- 8.9) mu mol/min mg. Antibody against the recombinant protein confirmed a glycosomal cellular localization of the enzyme in promastigotes. Data from light scattering and small angle X-ray scattering showed that a trimeric state is the active form of the protein. We determined empirically that a manganese wash at room temperature is the best condition to purify active enzyme. The interaction of the recombinant protein with the immobilized nickel also allowed us to confirm the structural disposition of histidine at positions 3 and 324. The determined structural parameters provide substantial data to facilitate the search for selective inhibitors of parasitic sources of arginase, which could subsequently point to a candidate for leishmaniasis therapy. (c) 2008 Elsevier B.V. All rights reserved.

Assessing the impact of deforestation and climate change on the range size and environmental niche of bird species in the Atlantic forests, Brazil

Aim Habitat loss and climate change are two major drivers of biological diversity. Here we quantify how deforestation has already changed, and how future climate scenarios may change, environmental conditions within the highly disturbed Atlantic forests of Brazil. We also examine how environmental conditions have been altered within the range of selected bird species. Location Atlantic forests of south-eastern Brazil. Methods The historical distribution of 21 bird species was estimated using Maxent. After superimposing the present-day forest cover, we examined the environmental niches hypothesized to be occupied by these birds pre- and post-deforestation using environmental niche factor analysis (ENFA). ENFA was also used to compare conditions in the entire Atlantic forest ecosystem pre- and post-deforestation. The relative influence of land use and climate change on environmental conditions was examined using analysis of similarity and principal components analysis. Results Deforestation in the region has resulted in a decrease in suitable habitat of between 78% and 93% for the Atlantic forest birds included here. Further, Atlantic forest birds today experience generally wetter and less seasonal forest environments than they did historically. Models of future environmental conditions within forest remnants suggest generally warmer conditions and lower annual variation in rainfall due to greater precipitation in the driest quarter of the year. We found that deforestation resulted in a greater divergence of environmental conditions within Atlantic forests than that predicted by climate change. Main conclusions The changes in environmental conditions that have occurred with large-scale deforestation suggest that selective regimes may have shifted and, as a consequence, spatial patterns of intra-specific variation in morphology, behaviour and genes have probably been altered. Although the observed shifts in available environmental conditions resulting from deforestation are greater than those predicted by climate change, the latter will result in novel environments that exceed temperatures in any present-day climates and may lead to biotic attrition unless organisms can adapt to these warmer conditions. Conserving intra-specific diversity over the long term will require considering both how changes in the recent past have influenced contemporary populations and the impact of future environmental change.

Using ecological niche modelling to infer past, present and future environmental suitability for Leiopelma hochstetteri, an endangered New Zealand native frog

Leiopelma hochstetteri is an endangered New Zealand frog now confined to isolated populations scattered across the North Island. A better understanding of its past, current and predicted future environmental suitability will contribute to its conservation which is in jeopardy due to human activities, feral predators, disease and climate change. Here we use ecological niche modelling with all known occurrence data (N = 1708) and six determinant environmental variables to elucidate current, pre-human and future environmental suitability of this species. Comparison among independent runs, subfossil records and a clamping method allow validation of models. Many areas identified as currently suitable do not host any known populations. This apparent discrepancy could be explained by several non exclusive hypotheses: the areas have not been adequately surveyed and undiscovered populations still remain, the model is over simplistic; the species` sensitivity to fragmentation and small population size; biotic interactions; historical events. An additional outcome is that apparently suitable, but frog-less areas could be targeted for future translocations. Surprisingly, pre-human conditions do not differ markedly highlighting the possibility that the range of the species was broadly fragmented before human arrival. Nevertheless, some populations, particularly on the west of the North Island may have disappeared as a result of human mediated habitat modification. Future conditions are marked with higher temperatures, which are predicted to be favourable to the species. However, such virtual gain in suitable range will probably not benefit the species given the highly fragmented nature of existing habitat and the low dispersal ability of this species. (C) 2010 Elsevier Ltd. All rights reserved.

Refolded dengue virus type 2 NS1 protein expressed in Escherichia coli preserves structural and immunological properties of the native protein

The dengue virus NS1 protein has been shown to be a protective antigen under different experimental conditions but the recombinant protein produced in bacterial expression systems is usually not soluble and loses structural and immunological features of the native viral protein In the present study, experimental conditions leading to purification and refolding of the recombinant dengue virus type 2 (DENV-2) NS1 protein expressed in Escherichia coil are described The refolded recombinant protein was recovered as heat-stable soluble dimers with preserved structural features, as demonstrated by spectroscopic methods In addition, antibodies against epitopes of the NS1 protein expressed in eukaryotic cells recognized the refolded protein expressed in E coli but not the denatured form or the same protein submitted to a different refolding condition Collectively, the results demonstrate that the recombinant NS1 protein preserved important conformation and antigenic determinants of the native virus protein and represents a valuable reagent either for the development of vaccines or for diagnostic methods. (C) 2010 Elsevier B V All rights reserved

Racial Conflict in the United States of America : A Deconstructive Perspective on Native Speaker by Changrae Lee

Written about the time of the Golden Venture incident, Chang-rae Lee’s Native Speaker makes a particular reference to that incident, whereby implying that particular immigrants, on the grounds of their racial identities, are mistreated and considered as aliens by some Americas. While some whites discriminate against immigrants, there is widespread ethnic tension between Korean Americans and African Americans. Significantly, racial conflict between Koreans and blacks and the racist attitude of some whites toward immigrants are mirrored in the relationship between the Korean-American protagonist Henry and his American wife Lelia. That is, due to their different racial identities they do not understand each other and they always argue. However, toward the end of the novel, Henry and Lelia come to understand each other. While ethnic conflict between Koreans and blacks and certain whites’ discriminatory attitudes toward immigrants is serious one, the novel suggests the unimportance of racial identity. In other words, the novel concludes that there is no discriminatory treatment of immigrants and, in fact, every one is a native Speaker in America. In the novel there is no message of how racial conflict could be resolved. However, this essay suggests that by investigating how the tension between Henry and Lelia is resolved, one could suggest a solution for the ethnicity problem in America and in real life.

Acoustic Variability in the Production of English Vowels by Native and Non-Native Speakers

The features of non-native speech which distinguish it from native speech are often difficult to pin down. It is possible to be a native speaker of any of a vast number of varieties of English. These varieties each have their phonetic characteristics which allow them to be identified by speakers of the varieties in question and by others. The phonetic differences between the accents represented by these varieties are very great. It is impossible to indicate any particular configuration of vowels in the acoustic vowel space or set of consonant articulations which all native-speaker varieties of English have in common and which non-native speakers do not share. This study considers the vowel quality in a single word by native and non-native speakers.

Relative durations of post-vocalic consonants in readaloud Spanish by native Swedish L2-learners

Prosody is said to be a persistent feature in foreign accents, particularly in the speech of adult learners. Native Swedish speakers have been reported to lengthen vowels in Spanish as well as post-vocalic consonants in English and German, more than native speakers of the respective languages. The aim of the present study is to examine whether native Swedish learners of Spanish produce increased postvocalic consonant durations in a reading aloud exercise. The text contains certain words that could be expected to trigger complementary consonant lengthening in native Swedish speakers. The result shows that there is no general tendency for native Swedes to lengthen post-vocalic consonants more than native speakers of Spanish in the present speech material. There are examples of longer consonant durations in the speech of the native,Swedish subjects, but it could be a coincidence. A conclusion is that the text used in the study, and probably Spanish as a language, contains few words that are ideal for triggering lengthening of post-vocalic consonants in native Swedish speakers.

November 2007: National American Indian & Alaska Native Heritage Month

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Processos produtivos e trabalho-educação: a incorporação do caboclo catarinense na indústria madeireira

The object theme of the present study is a population of caboclos that absorbed as manual workers in the saw-mills which were mounted in the highland region of Santa Catarina since 1950. The abundance of araucaria (native kind of pine) the opening of markets, and the corroboration of other industrial exploration conditions encouraged a great crowd of small en.- trepreneurs, coming basically from the state of Rio Grande do Sul, to migrate and settle down, building up a lot of saw-mills near rich forests and fields. The saw-mills started aprosperous production of timber sawn in planks. The process of industriali zation was so intensive, the destruction of pine woods soviolent that, in less than three decades, the forests ran out of tree reserves. The caboclos, absorbed as manual workers in production of timber, lived traditionally in an system of subsistense, either from the cultivation of pine the economic their land (planter caboclo) or as labourers in cattle-growing farms (farm hand caboclos). Nevertheless, the 'advantages' that were offered them by the new-comer entrepreneurs (a salary paid in money, a new house in a village, and other favours) helped the great majority of caboclos to abandon their traditional work and enlist as "workmen" in saw-mills. The new job, besides being a novelty, was an opportunity for a change in status. Subsequently, the running out of forests of araucaria and the resultant progressive shut-down of saw-mills caused the crowd of workmen to be out of imployment and to form to form a migratory flood toward the most important town of the region, Lages. The town of Lages, however, having made of the timber i ts main economic support wi thout the implantation of an alternative industry, was unable to offer the migrantssufficient 'work places'. In this way, the 'marginal crowd' began to settle down in the suburbs of the city. This study, in the context of the object theme, analyses two main questions related to the reality 'WOlLQ' and to the economic exploitation forms: ~) the relations of production in the economic regime of subsistence and in the capitalist regime of industrial production with the consequent 'positions' of the workman in the productive processj ~~) the deriving educative effects of the productive process, either in the economic regime of subsistence, or in the capitalist industrial regime. The two questions are theoretically debated andconfro~ ted with the proposed reality, giving origin to conclusions that, in a general formulation, can be summarized as follows: a) the caboclos of the highland region of Santa Catari na, when under an economic regime of subsistence, held in fee the productive processj there was a social division of the work and aclimate of freedom which made possible the development of knowledge from their life and work experience, the production of most of their tools, and the making of necessary manufactures adapted to their own surrounding ditionsi -- - --- other con- ------ b) however, these same caboclos, when absorbed by the capitalist industrial process of production - tipified by the work in saw-mills - lost the control of the productive processj this was caused by the technologic division of the work, since each man began to perform a dull and repeti tive action, directed by the speed of the 'major-saw' j man resigned form his skill and inventive power and surrendered to an executive authority which turned him into a 'collective worker'j the new productive process, besides rnaking each rnan a copy of a pattern, put the caboclos in a situation in which the daily work experiencedidn't add anything in terrns of autogenesis of knowledgei and even the environrnental educative rneans were reduced to new forrns of adaptation to the productive process, relegating rnan's inventive power to inertia.

Composição da fauna de formigas (Hymenoptera: Formicidae) de serapilheira em florestas semidecídua e de Eucalyptus spp., na região sudeste do Brasil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)