Vitellin-binding proteins in the nematode Oscheius tipulae (Nematoda, Rhabditida)

We describe the first application of a non-radioactive ligand-blotting technique to the characterization of proteins interacting with nematode vitellins. Chromatographically purified vitellins from the free-living nematode Oscheius tipulae were labeled with fluorescein in vitro. Ligand-blotting assays with horseradish peroxidase-conjugated anti-fluorescein antibodies showed that labeled vitellins reacted specifically with a polypeptide of approximately 100 kDa, which we named P100. This polypeptide is a specific worm`s vitellin-binding protein that is present only in adult worms. Blots containing purified O. tipulae vitellin preparations showed no detectable signal in the 100 kDa region, ruling out any possibility of yolk polypeptides self-assembling under the conditions used in our assay. Experiments done in the presence of alpha-methyl mannoside ruled out the possibility of vitellins binding to P100 through mannose residues. Triton X-114 fractionation of whole worm extracts showed that P100 is either a membrane protein or has highly hydrophobic regions. (C) 2008 Elsevier Inc. All rights reserved.

Oscheius tipulae as an example of eEF1A gene diversity in nematodes

We characterized four eEF1A genes in the alternative rhabditid nematode model organism Oscheius tipulae. This is twice the copy number of eEF1A genes in C. elegans, C. briggsae, and, probably, many other free-living and parasitic nematodes. The introns show features remarkably different from those of other metazoan eEF1A genes. Most of the introns in the eEF1A genes are specific to O. tipulae and are not shared with any of the other genes described in metazoans. Most of the introns are phase 0 (inserted between two codons), and few are inserted in protosplice sites (introns inserted between the nucleotide sequence A/CAG and G/A). Two of these phase 0 introns are conserved in sequence in two or more of the four eEF1A gene copies, and are inserted in the same position in the genes. Neither of these characteristics has been detected in any of the nematode eEF1A genes characterized to date. The coding sequences were also compared with other eEF1A cDNAs from 11 different nematodes to determine the variability of these genes within the phylum Nematoda. Parsimony and distance trees yielded similar topologies, which were similar to those created using other molecular markers. The presence of more than one copy of the eEF1A gene with nearly identical coding regions makes it difficult to define the orthologous cDNAs. As shown by our data on O. tipulae, careful and extensive examination of intron positions in the eEF1A gene across the phylum is necessary to define their potential for use as valid phylogenetic markers.

Regulation of Radial Glial Motility by Visual Experience

Radial glia in the developing optic tectum express the key guidance molecules responsible for topographic targeting of retinal axons. However, the extent to which the radial glia are themselves influenced by retinal inputs and visual experience remains unknown. Using multiphoton live imaging of radial glia in the optic tectum of intact Xenopus laevis tadpoles in conjunction with manipulations of neural activity and sensory stimuli, radial glia were observed to exhibit spontaneous calcium transients that were modulated by visual stimulation. Structurally, radial glia extended and retracted many filopodial processes within the tectal neuropil over minutes. These processes interacted with retinotectal synapses and their motility was modulated by nitric oxide (NO) signaling downstream of neuronal NMDA receptor (NMDAR) activation and visual stimulation. These findings provide the first in vivo demonstration that radial glia actively respond both structurally and functionally to neural activity, via NMDAR-dependent NO release during the period of retinal axon ingrowth.

A metodologia de análise de dados espaciais aplicada a plantas forrageiras

Dois experimentos e um levantamento por amostragem foram analisados no contexto de dados espaciais. Os experimentos foram delineados em blocos completos casualizados sendo que no experimento um (EXP 1) foram avaliados oito cultivares de trevo branco, sendo estudadas as variáveis Matéria Seca Total (MST) e Matéria Seca de Gramíneas (MSGRAM) e no experimento dois (EXP 2) 20 cultivares de espécies forrageiras, onde foi estudada a variável Percentagem de Implantação (%IMPL). As variáveis foram analisadas no contexto de modelos mistos, sendo modelada a variabilidade espacial através de semivariogramas exponencias, esféricos e gaussianos. Verificou-se uma diminuição em média de 19% e 14% do Coeficiente de Variação (CV) das medias dos cultivares, e uma diminuição em média de 24,6% e 33,3% nos erros padrões dos contrastes ortogonais propostos em MST e MSGRAM. No levantamento por amostragem, estudou-se a associação espacial em Aristida laevis (Nees) Kunth , Paspalum notatum Fl e Demodium incanum DC, amostrados em uma transecção fixa de quadros contiguos, a quatro tamanhos de unidades amostrais (0,1x0,1m; 0,1x0,3m; 0,1x0,5m; e 0,1x1,0m). Nas espécies Aristida laevis (Nees) Kunth e Paspalum notatum Fl, existiu um bom ajuste dos semivariogramas a tamanhos menores das unidades amostrais, diminuíndo quando a unidade amostral foi maior. Desmodium incanum DC apresentou comportamento contrario, ajustando melhor os semivariogramas a tamanhos maiores das unidades amostrais.

Combinação dos efeitos da carga animal com métodos físicos e químicos para o controle de plantas indesejáveis em pastagem nativa da Serra do Sudeste do RS

O experimento foi conduzido no município de Cachoeira do Sul de out/2001 a out/2002, objetivando avaliar a dinâmica vegetacional e o desempenho animal (31/05/02 a 16/09/02), em uma pastagem nativa submetida a diferentes métodos de controle de plantas indesejáveis. O local corresponde à região fisiográfica denominada Serra do Sudeste, com predominância das espécies Paspalum notatum (grama forquilha), Andropogon lateralis (capim caninha), além de Arachis burkartii e Desmodium incanum. Como plantas classificadas como indesejáveis encontram-se Baccharis trimera (carqueja), Vernonia nudiflora (alecrim) e Psidium luridum (araçá do campo). O delineamento experimental utilizado foi um fatorial em blocos (4 x 2), com duas repetições de campo. Os tratamentos foram os seguintes: testemunha (T); roçada de primavera (P); roçada de primavera + outono (O) e roçada de primavera + controle químico (Q), todos em duas ofertas de forragem, média (8%) e alta (14%). Os animais testes foram três novilhas por repetição, mantidas em pastejo contínuo com lotação variável. A massa de forragem foi obtida através de avaliações visuais usando padrões de referência, o acúmulo de forragem através do uso de gaiolas de exclusão e a composição botânica pelo método do ponto. Avaliaram-se os parâmetros de desempenho individual, carga animal, ganho de peso vivo por área e a dinâmica vegetacional. Os dados foram submetidos a análise de variância através do pacote estatístico SAS. O nível alto de oferta permitiu uma menor perda de peso vivo por área. Os tratamentos P e Q proporcionaram um melhor desempenho individual e menor perda de peso vivo por área, quando comparados a T. Os tratamentos P, O e Q foram eficientes no controle de Baccharis trimera. A. burkartii e Eriyngium horridum foram incrementados pela média intensidade de pastejo. Aristida laevis teve sua participação aumentada pela interação tratamento Q e baixa intensidade de pastejo.

Processos de degradação e regeneração da vegetação campestre do entorno de areais do sudoeste do Rio Grande do Sul

No Sudoeste do Rio Grande do Sul ocorrem manchas de substrato arenoso sem cobertura vegetal, conhecidos regionalmente como areais. A tese reexamina o problema e suas causas, enfatizando o papel da vegetação natural e propondo estratégias para a revegetação e prevenção à arenização em sistemas pastoris. Evidências foram obtidas em levantamentos e experimento avaliando processos de degradação e regeneração da vegetação campestre do entorno de areais. Levantamento da vegetação, realizado em 41 parcelas de 4,5 x 9,0 m na borda de 11 areais indicou a existência de dois tipos de comunidades. Areais de Manoel Viana e aqueles usados pelo gado em São Francisco de Assis, com alto percentual de substrato exposto, são caracterizados principalmente por Elyonurus sp., Axonopus pressus e Butia paraguayensis. Areais de Alegrete e aqueles excluídos de pastejo em São Francisco de Assis, com menor percentual de substrato exposto, são caracterizados principalmente por Andropogon lateralis e Aristida laevis. Foi também avaliada a dinâmica da vegetação em gradientes de arenização, usando quadros (0,25 m2) contíguos em 16 transecções de 10 m localizadas em cinco areais. A dinâmica da vegetação foi associada ao uso das áreas pelo gado, pois houve aumento, após 14 meses, de substrato exposto em comunidades de areais sob pastoreio, enquanto que aquelas sem gado apresentaram uma dinâmica espacial-temporal de maior estabilidade da cobertura vegetal. Um experimento foi realizado para avaliar, durante 8 meses, o efeito de níveis controlados de soterramento por areia (0, 5, 10 e 20 cm) em comunidades do entorno de dois areais sob pastoreio. Comunidades caracterizadas por Elyonurus sp. e Axonopus pressus foram mais tolerantes ao soterramento. As evidências indicam que a exclusão do gado de areais pode ser uma alternativa eficaz para a revegetação de areais por espécies das comunidades naturais do entorno. Ademais, a arenização pode ser prevenida pelo uso adequado dos campos que mantenha a cobertura vegetal natural protegendo o solo dos processos erosivos hídrico e eólico.

Caracterização e composição florística de uma comunidade savânica no Rio Grande do Norte, Brasil : subsídios para a conservação

Studies on Brazilian biodiversity are still very few and can observe the difference of knowledge between the different regions of the country. This affirmation can be verified in the investigation to identify the rare species in Brazil and the key biodiversity areas (ACBs). In that study were identified for Brasil 2.256 rare species and 752 ACBs. The Rio Grande do Norte (RN) was the only Brazilian state that has not been identified any rare species and no ACBs, possibly due to the lack of floristic studies in this state. A particular area was selected for this study: an area of ecological tension with savanna physiognomy in Rio do Fogo, RN. This savanna community is represented in RN in a fragment and immersed in restinga and caatinga and was identified and described only through of radar imagens there are no studies to date in loco . We have prepared the following questions about this community savanna: 1) The region delimited and described by through of radar images by the RADAMBRASIL, 1976, can be associated of the Cerrado, in terms of floristic ?; 2) What is the floristic composition of this area? This area includes rare species, endemic or endangered? 3) What is the geographical and phytogeographical distribution of plant species registered in this area? 4) Those plant species registered are endemic or have affinity with other areas phytoecological Brazilian? To answer these questions we performed a floristic inventory of the August 2007 to September/2009. The results are presented in two chapters (manuscripts). The first chapter, titled "The Savannah Rio Grande do Norte: floristic links with other plant formations in the Northeast and Center-West Brazil" was submitted to the Revista Brasileira de Botância. Chapter 1 discusses the phytogeographical distribution of the species, by comparing floristic studies conducted in the Cerrado, Caatinga and Restinga in the Northeast and Cerrado of the Central Brazil. The analysis of data of this study and compilation with other studies indicated that: i) the record of 94 plant species; ii) of total species, about 64% are associated with the Cerrado, the second specialized bibliography, and about 78% as the List of Species of Flora of Brazil. However, about 73% of total species (94) are also distributed in the Caatinga, the Atlantic forest 64%, the Amazon forest 64%, the Pantanal 15% and the Pampa 12%. Floristically the data show that the community studied is influenced by other floras, has a structure where grasses dominate and also because of his appearance the same savanna then be classified as a Savana gramíneo-lenhosa do tabuleiro . Chapter 2, titled "Considerations on the flora of a savanna community in Rio Grande do Norte, northeastern Brazil: Subsidy key area for conservation" was submitted to the Revista Natureza e Conservação. This has the objective of improve the knowledge of flora of Rio Grande do Norte and to identify possible rare species and consequently increase the key areas of biodiversity in Brazil. The data indicated that: i) of 94 species registered in the study area, 40 were new records for the Rio Grande do Norte state; ii) These citations to unpublished state, Stylosanthes montevidensis Vogel (Fabaceae) and Aristida laevis (Nees) Kunth (Poaceae) are indicated for the first time to the Northeast of Brazil; iii) are registered in the area 24 species endemic to Brazil and 63 non-endemic; iv) Aspilia procumbens Baker (Asteraceae) registered in the area is considered a restricted species and micro endemic Rio Grande do Norte, ie rare species; v) Aspilia procumbens is also cited in the category of critically endangered species and Stilpnopappus cearensis Hubber (Asteraceae) a species vulnerable to extinction. This study shows a new area phytoecological in Rio Grande do Norte and indicates the area's potential to contribute with the sites of global significance for biodiversity conservation, either locally, regionally and nationally. This will certainly contribute to respond some targets set by the Global Strategy for Plant Conservation and the Convention on Biological Diversity such as the inventory of vegetal diversity in a region with little collection, which will provide data that contributes to questions and themes related to biodiversity.

Structural basis for the specificity of bipartite nuclear localization sequence binding by importin-alpha

Importin-alpha is the nuclear import receptor that recognizes cargo proteins carrying conventional basic monopartite and bipartite nuclear localization sequences (NLSs) and facilitates their transport into the nucleus. Bipartite NLSs contain two clusters of basic residues, connected by linkers of variable lengths. To determine the structural basis of the recognition of diverse bipartite NLSs by mammalian importin-alpha, we co-crystallized a non-autoinhibited mouse receptor protein with peptides corresponding to the NLSs from human retinoblastoma protein and Xenopus laevis phosphoprotein N1N2, containing diverse sequences and lengths of the linker. We show that the basic clusters interact analogously in both NLSs, but the linker sequences adopt different conformations, whereas both make specific contacts with the receptor. The available data allow us to draw general conclusions about the specificity of NLS binding by importin-alpha and facilitate an improved definition of the consensus sequence of a conventional basic/bipartite NLS (KRX10-12KRRK) that can be used to identify novel nuclear proteins.

Contrasting responses to induction cues by ants inhabiting Maieta guianensis (Melastomataceae)

As duas formigas que mais comumente se associam com a mirmecofita de sub-bosque Maieta guianensis (Melastomataceac) respondem de maneiras diferentes a simulac (o) over tilde es experimentais de herbivoria. Enquanto Pheidole minutula & induzida tanto por danos fisicos e extratos de tecido foliar, Crematogaster laevis 6 induzida apenas pelos danos fisicos. Sugerimos que tal variac (a) over tildeo interespecifica em respostas induzidas e comum e pode influenciar a qualidade das defesas contra herbivoros que as formigas oferecem as plantas que elas ocupam.

Interspecific variation in the defensive responses of obligate plant-ants: experimental tests and consequences for herbivory

The aggressive behavior of ants that protect plants from herbivores in exchange for rewards such as shelter or food is thought to be an important form of biotic defense against herbivory, particularly in tropical systems. To date, however, no one has compared the defensive responses of different ant taxa associated with the same plant species, and attempted to relate these differences to longer-term efficacy of ant defense. We used experimental cues associated with herbivory-physical damage and extracts of chemical volatiles from leaf tissue-to compare the aggressive responses of two ant species obligately associated with the Amazonian myrmecophyte Tococa bullifera (Melastomataceae). We also conducted a colony removal experiment to quantify the level of resistance from herbivores provided to plants by each ant species. Our experiments demonstrate that some cues eliciting a strong response from one ant species elicited no response by the other. For cues that do elicit responses, the magnitude of these responses can vary interspecifically. These patterns were consistent with the level of resistance provided from herbivores to plants. The colony removal experiment showed that both ant species defend plants from herbivores: however, herbivory was higher on plants colonized by the less aggressive ant species. Our results add to the growing body of literature indicating defensive ant responses are stimulated by cues associated with herbivory. However, they also suggest the local and regional variation in the composition of potential partner taxa could influence the ecology and evolution of defensive mutualisms in ways that have previously remained unexplored.

Autonomic control of heart rate is virtually independent of temperature but seems related to the neuroanatomy of the efferent vagal supply to the heart in the bullfrog, Lithobathes catesbeianus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ant-tended hemiptera in Amazonian myrmecophytes: Patterns of abundance and implications for mutualism function (Hymenoptera: Formicidae)

We assessed how the abundance of ant-tended Hemiptera associated with two Amazonian myrmecophytes, Tococa bullifera and Maieta guianensis, varied as a function of resident ant species. We collected five species or morpho-species of adult hemiptera in the domatia of M. guianensis, with four of these species also found in Tococa bullifera. Maieta guianensis plants inhabited by Crematogaster laevis had over four-fold more hemiptera in them than plants inhabited by Pheidole minutula. In contrast, the density of hemiptera in Tococa bullifera domatia was independent of the species of ant resident. For each of the two ant species inhabiting Maieta guianensis, there was a positive and significant relationship between the abundance of hemiptera and workers inhabiting a plant. This relationship was also significant and positive for the Tococa bullifera plants inhabited by C. laevis. However, there was no relationship between Azteca worker and hemipteran density, although there was a trend towards a positive relationship. Our results indicate that hemipteran abundance can vary significantly between different myrmecophyte species, but that the nature of this relationship is mediated by the identity of the ant associate. Because hemipterans are herbivores, the costs and benefits of different ant partners to the host plant may vary in ways that are often overlooked.

Big, bad, and beautiful: phylogenetic relationships of the horned frogs (Anura: Ceratophryidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diversificação de anuros no Chaco Sul-Americano

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The breathing pattern and the ventilatory response to aquatic and aerial hypoxia and hypercarbia in the frog Pipa carvalhoi

Anuran amphibians are known to exhibit an intermittent pattern of pulmonary ventilation and to exhibit an increased ventilatory response to hypoxia and hypercarbia. However, only a few species have been studied to date. The aquatic frog Pipa carvalhoi inhabits lakes, ponds and marshes that are rich in nutrients but low in O-2. There are no studies of the respiratory pattern of this species and its ventilation during hypoxia or hypercarbia. Accordingly, the aim of the present study was to characterize the breathing pattern and the ventilatory response to aquatic and aerial hypoxia and hypercarbia in this species. With this purpose, pulmonary ventilation (V-1) was directly measured by the pneumotachograph method during normocapnic normoxia to determine the basal respiratory pattern and during aerial and aquatic hypercarbia (5% CO2) and hypoxia (5% O-2). Our data demonstrate that P. carvalhoi exhibits a periodic breathing pattern composed of single events (single breaths) of pulmonary ventilation separated by periods of apnea. The animals had an enhanced V-1 during aerial hypoxia, but not during aquatic hypoxia. This increase was strictly the result of an increase in the breathing frequency. A pronounced increase in V-1 was observed if the animals were simultaneously exposed to aerial and aquatic hypercarbia, whereas small or no ventilatory responses were observed during separately administered aerial or aquatic hypercarbia. P. carvalhoi primarily inhabits an aquatic environment. Nevertheless, it does not respond to low O-2 levels in water, although it does so in air. The observed ventilatory responses to hypercarbia may indicate that this species is similar to other anurans in possessing central chemoreceptors. (C) 2012 Elsevier Inc. All rights reserved.