Trypanosoma cruzi: establishment of permeable cells for RNA processing analysis with drugs

Pre-mRNA maturation in trypanosomatids occurs through a process called trans-splicing which involves excision of introns and union of exons in two independent transcripts. For the first time, we present the standardization of Trypanosoma cruzi permeable cells (Y strain) as a model for trans-splicing study of mRNAs in trypanosomes, following by RNase protection reaction, which localizes the SL exon and intron. This trans-splicing reaction in vitro was also used to analyze the influence of NFOH-121, a nitrofurazone-derivative, on this mechanism. The results suggested that the prodrug affects the RNA processing in these parasites, but the trans-splicing reaction still occurred.

The sample processing time interval as an influential factor in flow cytometry analysis of lymphocyte subsets

The objective of this paper is to propose a protocol to analyze blood samples in yellow fever 17DD vaccinated which developed serious adverse events. We investigated whether or not the time between sample collection and sample processing could interfere in lymphocyte subset percentage, for it is often impossible to analyze blood samples immediately after collection due to transport delay from collection places to the flow cytometry facility. CD4+CD38+ T, CD8+CD38+ T, CD3+ T, CD19+ B lymphocyte subsets were analyzed by flow cytometry in nine healthy volunteers immediately after blood collection and after intervals of 24 and 48 h. The whole blood lysis method and gradient sedimentation by Histopaque were applied to isolate peripheral blood mononuclear cells for flow cytometry analyses. With the lysis method, there was no significant change in lymphocyte subset percentage between the two time intervals (24 and 48 h). In contrast, when blood samples were processed by Histopaque gradient sedimentation, time intervals for sample processing influenced the percentage in T lymphocyte subsets but not in B cells. From the results obtained, we could conclude that the whole blood lysis method is more appropriate than gradient sedimentation by Histopaque for immunophenotyping of blood samples collected after serious adverse events, due to less variation in the lymphocyte subset levels with respect to the time factor.

Anti-mycobacterial treatment reduces high plasma levels of CXC-chemokines detected in active tuberculosis by cytometric bead array

Chemokines recruit and activate leukocytes, assisting granuloma formation. Herein, we evaluated plasma chemokines in patients with active tuberculosis (ATB) and after completing treatment (TTB) and compared them to BCG-vaccinated healthy controls (HC). Levels of chemokines were measured by cytometric bead array. Levels of CXCL8, CXCL9 and CXCL10 were higher in ATB patients compared to HC, but they decreased in TTB. Levels of CCL2 and CCL5 in ATB patients were similar to those observed in HC. Thus, the high levels of CXC-chemokines detected during ATB, which can modulate the trafficking of immune cells from the periphery to the site of infection, were reversed by anti-mycobacterial treatment.

Footprints of a trypanosomatid RNA world: pre-small subunit rRNA processing by spliced leader addition trans-splicing

The addition of a capped mini-exon [spliced leader (SL)] through trans-splicing is essential for the maturation of RNA polymerase (pol) II-transcribed polycistronic pre-mRNAs in all members of the Trypanosomatidae family. This process is an inter-molecular splicing reaction that follows the same basic rules of cis-splicing reactions. In this study, we demonstrated that mini-exons were added to precursor ribosomal RNA (pre-rRNA) are transcribed by RNA pol I, including the 5' external transcribed spacer (ETS) region. Additionally, we detected the SL-5'ETS molecule using three distinct methods and located the acceptor site between two known 5'ETS rRNA processing sites (A' and A1) in four different trypanosomatids. Moreover, we detected a polyadenylated 5'ETS upstream of the trans-splicing acceptor site, which also occurs in pre-mRNA trans-splicing. After treatment with an indirect trans-splicing inhibitor (sinefungin), we observed SL-5'ETS decay. However, treatment with 5-fluorouracil (a precursor of RNA synthesis that inhibits the degradation of pre-rRNA) led to the accumulation of SL-5'ETS, suggesting that the molecule may play a role in rRNA degradation. The detection of trans-splicing in these molecules may indicate broad RNA-joining properties, regardless of the polymerase used for transcription.

Digital soil mapping: strategy for data pre-processing

The region of greatest variability on soil maps is along the edge of their polygons, causing disagreement among pedologists about the appropriate description of soil classes at these locations. The objective of this work was to propose a strategy for data pre-processing applied to digital soil mapping (DSM). Soil polygons on a training map were shrunk by 100 and 160 m. This strategy prevented the use of covariates located near the edge of the soil classes for the Decision Tree (DT) models. Three DT models derived from eight predictive covariates, related to relief and organism factors sampled on the original polygons of a soil map and on polygons shrunk by 100 and 160 m were used to predict soil classes. The DT model derived from observations 160 m away from the edge of the polygons on the original map is less complex and has a better predictive performance.

Optical crop sensor for variable-rate nitrogen fertilization in corn: i - plant nutrition and dry matter production

Variable-rate nitrogen fertilization (VRF) based on optical spectrometry sensors of crops is a technological innovation capable of improving the nutrient use efficiency (NUE) and mitigate environmental impacts. However, studies addressing fertilization based on crop sensors are still scarce in Brazilian agriculture. This study aims to evaluate the efficiency of an optical crop sensor to assess the nutritional status of corn and compare VRF with the standard strategy of traditional single-rate N fertilization (TSF) used by farmers. With this purpose, three experiments were conducted at different locations in Southern Brazil, in the growing seasons 2008/09 and 2010/11. The following crop properties were evaluated: above-ground dry matter production, nitrogen (N) content, N uptake, relative chlorophyll content (SPAD) reading, and a vegetation index measured by the optical sensor N-Sensor® ALS. The plants were evaluated in the stages V4, V6, V8, V10, V12 and at corn flowering. The experiments had a completely randomized design at three different sites that were analyzed separately. The vegetation index was directly related to above-ground dry matter production (R² = 0.91; p<0.0001), total N uptake (R² = 0.87; p<0.0001) and SPAD reading (R² = 0.63; p<0.0001) and inversely related to plant N content (R² = 0.53; p<0.0001). The efficiency of VRF for plant nutrition was influenced by the specific climatic conditions of each site. Therefore, the efficiency of the VRF strategy was similar to that of the standard farmer fertilizer strategy at sites 1 and 2. However, at site 3 where the climatic conditions were favorable for corn growth, the use of optical sensors to determine VRF resulted in a 12 % increase in N plant uptake in relation to the standard fertilization, indicating the potential of this technology to improve NUE.

Optical crop sensor for variable-rate nitrogen fertilization in corn: II - indices of fertilizer efficiency and corn yield

Generally, in tropical and subtropical agroecosystems, the efficiency of nitrogen (N) fertilization is low, inducing a temporal variability of crop yield, economic losses, and environmental impacts. Variable-rate N fertilization (VRF), based on optical spectrometry crop sensors, could increase the N use efficiency (NUE). The objective of this study was to evaluate the corn grain yield and N fertilization efficiency under VRF determined by an optical sensor in comparison to the traditional single-application N fertilization (TSF). With this purpose, three experiments with no-tillage corn were carried out in the 2008/09 and 2010/11 growing seasons on a Hapludox in South Brazil, in a completely randomized design, at three different sites that were analyzed separately. The following crop properties were evaluated: aboveground dry matter production and quantity of N uptake at corn flowering, grain yield, and vegetation index determined by an N-Sensor® ALS optical sensor. Across the sites, the corn N fertilizer had a positive effect on corn N uptake, resulting in increased corn dry matter and grain yield. However, N fertilization induced lower increases of corn grain yield at site 2, where there was a severe drought during the growing period. The VRF defined by the optical crop sensor increased the apparent N recovery (NRE) and agronomic efficiency of N (NAE) compared to the traditional fertilizer strategy. In the average of sites 1 and 3, which were not affected by drought, VRF promoted an increase of 28.0 and 41.3 % in NAE and NRE, respectively. Despite these results, no increases in corn grain yield were observed by the use of VRF compared to TSF.

Determinação da tensão de água em solo agrícola usando um sensor de dissipação de calor

Um estudo foi realizado sob condições de laboratório e de campo objetivando avaliar um sensor de solo utilizado para determinação da temperatura e da tensão da água no solo. Devido à solubilidade do gesso, material poroso utilizado na fabricação dos sensores, houve redução em área da seção transversal e volume, e aumento da porosidade efetiva desses sensores, em relação aos valores observados antes do início dos testes. Os valores médios da temperatura observados com os sensores foram praticamente iguais aos observados com um termômetro de referência com precisão de 0,1°C. Quando sensores saturados foram instalados a 5 cm de profundidade em um solo franco-argilo-siltoso secado ao ar, em relação a valores de Delta T, o tempo de resposta do sensor à secagem foi um pouco superior a 24 horas. Valores de Delta T observados, tanto no laboratório quanto no campo, apresentaram maior variabilidade na faixa representativa de solo seco, em comparação com o solo úmido. Sob condições de campo, não houve diferença estatística (teste F; 5%) entre as médias de Delta T obtidas com os sensores de dissipação de calor instalados a 10 cm de profundidade e as médias de tensão obtidas com tensiômetros e transformadas para Delta T.

Imagens do sensor MODIS associadas a um modelo agronômico para estimar a produtividade de soja

O objetivo deste trabalho foi estimar a produtividade de soja no Rio Grande do Sul, nas safras de 2000/2001 a 2002/2003, por meio de um modelo agronômico implementado em um Sistema de Informação Geográfica (SIG). Duas abordagens foram utilizadas: o modelo agronômico (AGRO), com valores de índice de área foliar (IAF) obtidos da literatura; e o modelo agronômico-espectral (AGROESPEC), com valores de IAF estimados a partir das imagens MODIS (moderate resolution imaging spectroradiometer). As estimativas de produtividade obtidas pelo modelo foram comparadas àquelas fornecidas pelo Instituto Brasileiro de Geografia e Estatística (IBGE), com o uso do teste t para pares de observação. Nas safras 2000/2001 e 2001/2002, não foram observadas diferenças significativas. Para 2002/2003, o modelo subestimou o valor de produtividade em 7,87 e 7,04%, nas abordagens AGRO e AGROESPEC, respectivamente, em comparação à produtividade fornecida pelo IBGE. Ambas as abordagens do modelo permitiram avaliação objetiva e quantitativa do efeito das condições meteorológicas sobre a produtividade de soja. Entretanto, o AGROESPEC forneceu estimativas mais detalhadas, no que se refere à variação espacial da produtividade, em razão do emprego dos valores de IAF estimados a partir das imagens MODIS.

Mapeamento da temperatura da superfície terrestre com uso do sensor AVHRR/NOAA

O objetivo deste trabalho foi avaliar a adequação do uso do sensor AVHRR/NOAA (Advanced Very High Resolution Radiometer/National Oceanic and Atmospheric Administration) para mapeamento da temperatura da superfície terrestre (TST) no Estado do Rio Grande do Sul, por meio da comparação entre três algoritmos clássicos. Foram comparados os métodos de Becker & Li, Sobrino et al. e Kerr et al. para estimativa das TST mínimas, utilizando imagens noturnas e logo após o amanhecer. Os dados de emissividade e TST foram obtidos por meio de combinações matemáticas da radiação detectada nas faixas do visível, infravermelho próximo e termal do sensor AVHRR/NOAA. O sensor AVHRR é adequado para o mapeamento de TST para as condições do tipo de cobertura do solo que predominam no Rio Grande do Sul, visto que a TST estimada pelos três métodos testados foi próxima à temperatura do ar medida nos locais selecionados. O método de Sobrino et al. foi o mais adequado.

Comportamento do NDVI obtido por sensor ótico ativo em cereais

O objetivo deste trabalho foi avaliar, com um sensor ótico ativo, o comportamento do índice de vegetação por diferença normalizada (NDVI - "normalized difference vegetation index"), nas culturas de trigo, triticale, cevada e milho. Cinco experimentos foram conduzidos no Paraná e São Paulo, com variação de classes de solo, doses e fontes de N, e variedades de trigo. As seguintes variáveis foram avaliadas: NDVI, teor de N foliar, matéria seca e produtividade das culturas. Análises de regressões foram realizadas entre as doses de N aplicadas e NDVI, teor de N foliar, matéria seca e produtividade. Análises de correlação entre as variáveis foram realizadas. O trigo, triticale e cevada apresentaram resposta às aplicações de doses crescentes de N, pelo aumento nas leituras do NDVI, no teor de N foliar e na produtividade. Medido pelo sensor ótico ativo utilizado, o NDVI apresenta alto potencial para manejo do N nas culturas do trigo, triticale e cevada, e baixo potencial para a cultura do milho. Há interferência das variedades de trigo nas leituras do sensor ótico ativo.

Chemical and physical composition of grain-type and food-type soybean for food processing

The objective of this work was to evaluate the chemical and physical characteristics of grains of soybean (Glycine max) cultivars for food processing. The soybean cultivars evaluated were: grain-type - BRS 133 and BRS 258; food-type - BRS 213 (null lipoxygenases), BRS 267 (vegetable-type) and BRS 216 (small grain size). BRS 267 and BRS 216 cultivars showed higher protein content, indicating that they could promote superior nutritional value. BRS 213 cultivar showed the lowest lipoxygenase activity, and BRS 267, the lowest hexanal content. These characteristics can improve soyfood flavor. After cooking, BRS 267 cultivar grains presented a higher content of aglycones (more biologically active form of isoflavones) and oleic acid, which makes it proper for functional foods and with better stability for processing, and also showed high content of fructose, glutamic acid and alanine, compounds related to the soybean mild flavor. Because of its large grain size, BRS 267 is suitable for tofu and edamame, while small-grain-sized BRS 216 is good for natto and for soybean sprouts production. BRS 216 and BRS 213 cultivars presented shorter cooking time, which may be effective for reducing processing costs.

Avaliação de fluxos de calor e evapotranspiração pelo modelo SEBAL com uso de dados do sensor ASTER

O objetivo deste trabalho foi avaliar a eficiência da aplicação do modelo SEBAL em estimar os fluxos de energia em superfície e a evapotranspiração diária, numa extensa área de cultivo de arroz irrigado, no município de Paraíso do Sul, RS, tendo como parâmetros dados do sensor ASTER. As variáveis estudadas constituem importantes parâmetros do tempo e do clima em estudos agrometeorológicos e de racionalização no uso da água. As metodologias convencionais de estimativa desses parâmetros são pontuais e geralmente apresentam incertezas, que aumentam quando o interesse é o comportamento espacial desses parâmetros. Aplicou-se o algoritmo "Surface Energy Balance Algorithm for Land" (SEBAL), em uma imagem do sensor "Advanced Spaceborne Thermal Emission and Reflection Radiometer" (ASTER). As estimativas obtidas foram comparadas com medições em campo, realizadas por uma estação micrometeorológica localizada no interior da área de estudo. As estimativas mais precisas foram as de fluxo de calor sensível e de evapotranspiração diária, e a estimativa que apresentou maior erro foi a do fluxo de calor no solo. A metodologia empregada foi capaz de reproduzir os fluxos de energia em superfície de maneira satisfatória para estudos agrometeorológicos e de rendimento de culturas.

Validação de modelo para predição do potencial produtivo de trigo com sensor óptico ativo

O objetivo deste trabalho foi validar um modelo baseado no índice de vegetação por diferença normalizada (NDVI), para a predição da variabilidade espacial da biomassa acumulada na parte aérea e da produtividade de grãos de trigo. Foram realizadas leituras georreferenciadas de NDVI e biomassa, no final do afilhamento, e produtividade. O modelo proposto permitiu relacionar classes de variabilidade espacial do NDVI e da biomassa, com 81% de correspondência entre elas. Entretanto, a relação entre as classes de variabilidade espacial da produtividade e do NDVI foi menor, com 48% de correspondência.

Changes in the isoflavone profile and in the chemical composition of tempeh during processing and refrigeration

The objective of this work was to analyze changes in the isoflavone profile, determined by high performance liquid chromatography, at different processing stages and after refrigeration of tempeh. For tempeh production, clean soybean grains from cultivars BR 36 (low isoflavone content) and IAS 5 (high) were dehulled, and the separated cotyledons were hydrated and then cooked in boiling water for 30 min. Spores of the fungus Rhizopus microsporus var. oligosporus were inoculated in the cooked and cooled cotyledons, and incubated at 32ºC for 6, 12, 18, and 24 hours in perforated polypropylene bags, for fermentation. The resulting tempeh was stored at 4ºC for 6, 12, 18, and 24 hours. After 24-hour fermentation, isoflavone glucosides were 50% reduced, and the aglycone forms in the tempeh from both cultivars was increased. The malonyl forms reduced 83% after cooking. Less than 24 hours of refrigeration did not affect the isoflavone profile of tempeh from either cultivar, which is a good indicator of its quality. The tempeh maintains the high and low isoflavone content of the cultivars, which indicates that cultivar differences in this trait should be considered when processing tempeh.