Differential tissue tropism of Trypanosoma cruzi strains: an in vitro study

We have previously demonstrated selection favoring the JG strain of Trypanosoma cruziin hearts of BALB/c mice that were chronically infected with an equal mixture of the monoclonal JG strain and a clone of the Colombian strain, Col1.7G2. To evaluate whether cell invasion efficiency drives this selection, we infected primary cultures of BALB/c cardiomyocytes using these same T. cruzi populations. Contrary to expectation, Col1.7G2 parasites invaded heart cell cultures in higher numbers than JG parasites; however, intracellular multiplication of JG parasites was more efficient than that of Col1.7G2 parasites. This phenomenon was only observed for cardiomyocytes and not for cultured Vero cells. Double infections (Col1.7G2 + JG) showed similar results. Even though invasion might influence tissue selection, our data strongly suggest that intracellular development is important to determine parasite tissue tropism.

Immunostimulatory property of a synthetic peptide belonging to the soluble ATP diphosphohydro-lase isoform (SmATPDase 2) and immunolocalisation of this protein in the Schistosoma mansoni egg

A peptide (SmB2LJ; r175-194) that belongs to a conserved domain from Schistosoma mansoni SmATPDase 2 and is shared with potato apyrase, as predicted by in silico analysis as antigenic, was synthesised and its immunostimulatory property was analysed. When inoculated in BALB/c mice, this peptide induced high levels of SmB2LJ-specific IgG1 and IgG2a subtypes, as detected by enzyme linked immunosorbent assay. In addition, dot blots were found to be positive for immune sera against potato apyrase and SmB2LJ. These results suggest that the conserved domain r175-194 from the S. mansoni SmATPDase 2 is antigenic. Western blots were performed and the anti-SmB2LJ antibody recognised in adult worm (soluble worm antigen preparation) or soluble egg antigen antigenic preparations two bands of approximately 63 and 55 kDa, molecular masses similar to those predicted for adult worm SmATPDase 2. This finding strongly suggests the expression of this same isoform in S. mansoni eggs. To assess localisation of SmATPDase 2, confocal fluorescence microscopy was performed using cryostat sections of infected mouse liver and polyclonal antiserum against SmB2LJ. Positive reactions were identified on the external surface from the miracidium in von Lichtenberg's envelope and, in the outer side of the egg-shell, showing that this soluble isoform is secreted from the S. mansoni eggs.

Differential in vitro activity of the DNA topoisomerase inhibitor idarubicin against Trypanosoma rangeli and Trypanosoma cruzi

In this study the effect of eight DNA topoisomerase inhibitors on the growth Trypanosoma rangeli epimastigotes in cell culture was investigated. Among the eight compounds tested, idarubicin was the only compound that displayed promising trypanocidal activity with a half-maximal growth inhibition (GI50) value in the sub-micromolar range. Fluorescence-activated cell sorting analysis showed a reduction in DNA content in T. rangeli epimastigotes when treated with idarubicin. In contrast to T. rangeli, against Trypanosoma cruzi epimastigotes idarubicin was much less effective exhibiting a GI50 value in the mid-micromolar range. This result indicates that idarubicin displays differential toxic effects in T. rangeli and T. cruzi. Compared with African trypanosomes, it seems that American trypanosomes are generally less susceptible to DNA topoisomerase inhibitors.

Differential expression of the costimulatory molecules CD86, CD28, CD152 and PD-1 correlates with the host-parasite outcome in leprosy

Leprosy is a spectral disease exhibiting two polar sides, namely, lepromatous leprosy (LL) characterised by impaired T-cell responses and tuberculoid leprosy in which T-cell responses are strong. Proper T-cell activation requires signalling through costimulatory molecules expressed by antigen presenting cells and their ligands on T-cells. We studied the influence of costimulatory molecules on the immune responses of subjects along the leprosy spectrum. The expression of the costimulatory molecules was evaluated in in vitro-stimulated peripheral blood mononuclear cells of lepromatous and tuberculoid patients and healthy exposed individuals (contacts). We show that LL patients have defective monocyte CD86 expression, which likely contributes to the impairment of the antigen presentation process and to patients anergy. Accordingly, CD86 but not CD80 blockade inhibited the lymphoproliferative response to Mycobacterium leprae. Consistent with the LL anergy, there was reduced expression of the positive signalling costimulatory molecules CD28 and CD86 on the T-cells in these patients. In contrast, tuberculoid leprosy patients displayed increased expression of the negative signalling molecules CD152 and programmed death-1 (PD-1), which represents a probable means of modulating an exacerbated immune response and avoiding immunopathology. Notably, the contacts exhibited proper CD86 and CD28 expression but not exacerbated CD152 or PD-1 expression, suggesting that they tend to develop a balanced immunity without requiring immunosuppressive costimulatory signalling.

Immunologic evaluation and validation of methods using synthetic peptides derived from Mycobacterium tuberculosis for the diagnosis of tuberculosis infection

The goal of this study was to demonstrate the usefulness of an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of pulmonary tuberculosis (PTB) and extrapulmonary TB (EPTB). This assay used 20 amino acid-long, non-overlapped synthetic peptides that spanned the complete Mycobacterium tuberculosis ESAT-6 and Ag85A sequences. The validation cohort consisted of 1,102 individuals who were grouped into the following five diagnostic groups: 455 patients with PTB, 60 patients with EPTB, 40 individuals with non-EPTB, 33 individuals with leprosy and 514 healthy controls. For the PTB group, two ESAT-6 peptides (12033 and 12034) had the highest sensitivity levels of 96.9% and 96.2%, respectively, and an Ag85A-peptide (29878) was the most specific (97.4%) in the PTB groups. For the EPTB group, two Ag85A peptides (11005 and 11006) were observed to have a sensitivity of 98.3% and an Ag85A-peptide (29878) was also the most specific (96.4%). When combinations of peptides were used, such as 12033 and 12034 or 11005 and 11006, 99.5% and 100% sensitivities in the PTB and EPTB groups were observed, respectively. In conclusion, for a cohort that consists entirely of individuals from Venezuela, a multi-antigen immunoassay using highly sensitive ESAT-6 and Ag85A peptides alone and in combination could be used to more rapidly diagnose PTB and EPTB infection.

Primary dengue virus infections induce differential cytokine production in Mexican patients

Severe dengue pathogenesis is not fully understood, but high levels of proinflammatory cytokines have been associated with dengue disease severity. In this study, the cytokine levels in 171 sera from Mexican patients with primary dengue fever (DF) and dengue haemorrhagic fever (DHF) from dengue virus (DENV) 1 (n = 116) or 2 (n = 55) were compared. DF and DHF were defined according to the patient’s clinical condition, the primary infections as indicated by IgG enzymatic immunoassay negative results, and the infecting serotype as assessed by real-time reverse transcription-polymerase chain reaction. Samples were analysed for circulating levels of interleukin (IL)-12p70, interferon (IFN)-γ, tumour necrosis factor (TNF)-α, IL-6, and IL-8 using a commercial cytometric bead array. Significantly higher IFN-γ levels were found in patients with DHF than those with DF. However, significantly higher IL-12p70, TNF-α, and IL-6 levels were associated with DHF only in patients who were infected with DENV2 but not with DENV1. Moreover, patients with DF who were infected with DENV1 showed higher levels of IL-12p70, TNF-α, and IL-6 than patients with DHF early after-fever onset. The IL-8 levels were similar in all cases regardless of the clinical condition or infection serotype. These results suggest that the association between high proinflammatory cytokine levels and dengue disease severity does not always stand, and it once again highlights the complex nature of DHF pathogenesis.

The use of expert systems on the differential diagnosis of urinary incontinence

The differential diagnosis of urinary incontinence classes is sometimes difficult to establish. As a rule, only the results of urodynamic testing allow an accurate diagnosis. However, this exam is not always feasible, because it requires special equipment, and also trained personnel to lead and interpret the exam. Some expert systems have been developed to assist health professionals in this field. Therefore, the aims of this paper are to present the definition of Artificial Intelligence; to explain what Expert System and System for Decision Support are and its application in the field of health and to discuss some expert systems for differential diagnosis of urinary incontinence. It is concluded that expert systems may be useful not only for teaching purposes, but also as decision support in daily clinical practice. Despite this, for several reasons, health professionals usually hesitate to use the computer expert system to support their decision making process.

Adaptations of two specialist herbivores to movement on the hairy leaf surface of their host, Solanum guaraniticum Hassl (Solanaceae)

Plant trichomes can difficult the attachment and movement of small insects. Here, we examine the hypothesis that the success on the use of densely haired hosts by two cassidine species is determined by differential morphology and behavior. Larvae of Gratiana graminea (Klug, 1829) and Gratiana conformis (Boheman, 1854) move on the leaf surface of their host, Solanum guaraniticum Hassl by anchoring their tarsungulus on the trichome rays or by inserting the tarsungulus tip directly into epidermis. This kind of movement is only possible due to a similar tarsungulus shape among the species. Tarsungulus growth pattern is also similar between species, being relatively small on the posterior aperture, matching the diameter of the host plant trichome rays. The tarsungulus shape associated with differences on ontogenetic growth and attachment pattern allow these two Cassidinae larvae to efficiently move on the pubescent leaf surface of their host.

Estimativa do teor de água no solo utilizando dados de radar de abertura sintética

Em virtude da crescente demanda mundial por alimentos, um monitoramento eficaz e em larga escala da umidade do solo constitui fator de grande importância para a previsão de safras. Este trabalho teve por objetivo apresentar uma técnica para o cálculo do teor de água no solo, utilizando modelos preditivos de umidade do solo, baseados em dados de radar de abertura sintética (SAR). Foram utilizados dados do SAR a bordo do JERS-1 ("Japanese Earth Resources Satellite") e dois modelos empíricos. O primeiro relaciona o coeficiente de retroespalhamento com a permissividade complexa (modelo de Dubois), e o segundo relaciona a permissividade complexa com o teor de água do solo e algumas de suas características físico-hídricas, tais como percentagem de areia e argila (modelo de Hallikainen). Inicialmente, os dados do SAR/JERS-1 foram calibrados e, por meio do modelo de Dubois, foram calculados os valores de permissividade complexa. Para tanto, foi necessário inserir níveis estimados de rugosidade do solo. A partir destes resultados, utilizou-se o modelo de Hallikainen para calcular a umidade volumétrica. A análise geral dos resultados indica que a técnica de estimação de umidade do solo a partir de imagens de radar de abertura sintética, utilizada neste estudo, mostrou-se física e matematicamente exeqüível. No entanto, apresentou uma precisão moderada, não sendo ainda recomendada para o uso operacional no mapeamento de umidade do solo. A análise dos resultados revelou também que a precisão dos dados é bastante influenciada pela precisão dos valores de rugosidade introduzidos.

Uso do radar penetrante no solo (GPR) na investigação dos solos dos tabuleiros costeiros no litoral norte do estado da Bahia

Foram estudados nove perfis ao longo de uma toposseqüência sobre os sedimentos do Grupo Barreiras, na Fazenda Rio Negro, município de Entre Rios (BA), utilizando a prospecção eletromagnética por meio do Radar Penetrante no Solo - "Ground-penetrating radar - GPR", objetivando analisar a utilização dessa ferramenta na aquisição de informações sobre as feições que ocorrem no solo, mediante a comparação entre os radargramas obtidos e a descrição pedológica. O equipamento utilizado foi um Geophysical Survey System modelo GPR SR system-2, com antena de 80 MHz. A análise radargramétrica confirmou o aparecimento dos fragipãs e duripãs em profundidade, que ocorrem sempre acompanhados de um processo de transformação dos solos do tipo Latossolo Amarelo e Argissolo Amarelo em Espodossolo. Os padrões de reflexão mostram claramente os domínios dos solos argilosos e dos solos arenosos, com e sem a presença dos horizontes endurecidos.

Activity of phosphorus of synthetic Fe-K-P compounds in superphosphate fertilizers as affected by pH and ionic strength

The concentration of orthophosphate ions released from Fe-K-P compounds (Fe3KH8(PO4)6 .6H2O and Fe3KH14(PO4)8 .4H2O) present in superphosphates increases with pH, which initially suggests that the agronomic effectiveness of P fertilizers containing high amounts of these compounds would also increase with soil pH but studies considering activity, instead of concentration, are necessary. With this purpose, both compounds were synthesized under laboratory conditions, characterized by elemental chemical analysis, optical microscopy, X ray diffractometry, scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS), and used in a solubility study. Solutions of 0.01, 0.05 and 0.1 mol L-1 NaCl with pH adjusted to 3.0, 4.0, 5.0, 5.5, 6.0, 6.5, 7.0 and 7.5 were prepared for the solubility study of H8-syn, H14-syn and a phosphate rock (PR) from Brazil. The orthophosphate activity as H2PO4- and HPO4(2-) was calculated in each situation as related to pH and ionic strength using software MINTEQ. The remaining precipitates after equilibrium were chemically analyzed and subjected to X ray, SEM and EDS. Results of chemical analysis and instrumental techniques confirmed the preparation method. The activity of orthophosphate ions of both compounds tended to decrease under increasing pH and/or ionic strength of the solution, which in turn suggests that an increase in the solution pH does not necessarily promote an increase in the P bioavailability for plant uptake. This can be important when evaluating agronomic data of P fertilizers with high contents of these two Fe-K-P compounds.

Cristallochemical characterization of synthetic Zn-substituted maghemites (g-Fe2-xZn xO3)

Maghemite (g-Fe2O3) is the most usually found ferrimagnetic oxide in red basalt-derived soils. The variable degrees of ionic substitution of Fe3+ for different metals (e.g. Ti4+, Al3+, Mg2+, Zn2+, and Mn2+) and non-metals in the maghemite structure influence some cristallochemical features of this iron oxide. In this study, synthetic Zn-substituted maghemites were prepared by co-precipitation in alkaline aqueous media of FeSO4.7H2O with increasing amounts of ZnSO4.7H2O to obtain the following sequence of Fe3+ for Zn2+ substitutions: 0.0, 0.025, 0.05, 0.10, 0.15, 0.20, and 0.30 mol mol-1. The objective of this work was to evaluate the cristallochemical alterations of synthetic Zn-substituted maghemites. The dark black synthetic precipitated material was heated to 250 °C during 4 h forming a brownish maghemite that was characterized by chemical analysis as well as X ray diffraction (XRD), specific surface area and mass-specific magnetic susceptibility. The isomorphic substitution levels observed were of 0.0013, 0.0297, 0.0590, 0.1145, 0.1764, 0.2292 and 0.3404 mol mol-1, with the formation of a series of maghemites from Fe2Zn0O3 to Fe(1.49)Zn(0.770)O3 . The increase in Fe3+ for Zn2+ substitution, [Zn mol mol-1] increased the dimension a0 of the cubic unit cells of the studied maghemites according to the regression equation: a0 = 0.8343 + 0.02591Zn (R² = 0.98). On the other hand, the mean crystallite dimension and mass-specific magnetic susceptibility of the studied maghemites decreased with increasing isomorphic substitution.

Uso de sistema GPR (Ground Penetring Radar) na avaliação de atributos de um solo sob plantio de cana-de-açúcar

O GPR utiliza técnica indireta para investigação das estruturas de subsuperfície, sendo caracterizado como método não-invasivo, o qual permite extrair informações ao longo do perfil de solo sem perfurar, sondar ou escavar. Este trabalho teve o objetivo de avaliar a utilização do GPR no estabelecimento de atributos físicos do solo em área de cultivo com cana-de-açúcar, além de estabelecer sua correlação com o método do cilindro volumétrico. A área de estudo está localizada no município de Bom Sucesso - PR, sendo utilizada com cultura da cana há mais de 20 anos. Foram obtidas amostras de solos indeformadas com cilindro volumétrico nas profundidades de 0-0,1, 0,1-0,2, 0,2-0,3, 0,3-0,4 e 0,4-0,5 m, ao longo de três transeptos paralelos, sendo o central referente à linha de plantio (transepto 2) e os outros dois correspondentes às entrelinhas laterais (transeptos 1 e 3). Os valores de densidade e porosidade total, obtidos por meio das amostras físicas, foram inicialmente interpretados para verificar o estado do solo nas linhas e nas entrelinhas de cana, nas cinco profundidades. Posteriormente, foram correlacionados com os valores das constantes dielétricas (K) adquiridos pelo georradar, porém apenas para a camada de 0,4-0,5 m, uma vez que as camadas superficiais até a profundidade de 0,4 m não puderam ser avaliadas devido às limitações técnicas do equipamento, aliadas às condições físicas e químicas do solo, que acabaram por prejudicar as reflexões. A densidade mostrou-se menor nas linhas, quando comparada às entrelinhas, para as camadas de 0-0,1 e 0,2-0,3 m. A porosidade total foi menor nas entrelinhas do que nas linhas na camada de 0-0,1 m. Os valores de K foram maiores nas linhas, onde a umidade e a porosidade total foram maiores, e menores nas entrelinhas, onde as densidades encontravam se próximas ao limite crítico. A correlação de K com a densidade e porosidade total não foi significativa. Pequenas modificações na porosidade do solo em função do aumento da densidade foram suficientes para diminuir o valor de K nas entrelinhas, devido à redução na quantidade de água nos poros. Esse fato pode ser confirmado pela correlação de K com a umidade. Assim, pode-se dizer que o georradar foi extremamente sensível à variação de umidade no solo, apresentando melhores resultados para esta variável do que para a densidade e porosidade total determinada por meio do cilindro volumétrico.

Differential Behavior of Young Eucalyptus Clones in Response to Nitrogen Supply

Eucalyptus requires large amounts of nitrogen (N); however, it responds in diverse manners to the application of this nutrient. The aim of this study was to evaluate the differential performance in growth, mineral nutrition, and gas exchanges of N-fertilized Eucalyptus clones. The treatments consisted of two Eucalyptus clones (VM-01 and I-144) and six N application rates (0, 0.74, 2.93, 4.39, 5.85, and 8 mmol L-1 NH4NO3) arranged in a randomized complete block design with five replications. VM-01 had greater plant height and greater height/collar diameter ratio, as well as higher leaf concentrations of all macronutrients and of Cu, Fe, Mo, and Zn. In terms of total and root dry matter production, root/shoot ratio, and collar diameter, as well as stomatal conductance and transpiration, I-144 performed better. The performance of the clones was clearly differentiated, and the growth of I-144, despite lower leaf N concentration, was in general better than VM-01.

Differential vegetative and reproductive performances among fifteen guinea grass hybrids

The main scope of this work was to detect (Panicum maximum Jacq.) genotype differences as to morphoagronomic and seed quality indices, and to establish character correlations useful for determining vegetative and reproductive trends. Besides the flowering cycle, eight phenological and two seed quality traits were scored in a greenhouse randomized complete block experiment, as follows: plant height (PH), reproductive tiller number/overall tiller number (RTN/OTN), panicle number/reproductive tillers (PN/RT), leaf length (LL), leaf width (LW), panicle length (PL), fresh weight (FW), dry weight (DW), number of seeds/g (NS/G) and seed sample physical purity (SPP). Very-early and early-flowering hybrids consistently showed the highest correlation values among flowering cycle and RTN/OTN (r = -0.59**), PN/RT (r = -0.48**), NS/G (r = -0.88**) and SPP (r = -0.80**) (reproductive parameters) while intermediate and late-flowering hybrids presented the highest values for LL (r = 0.53**), LW (r = 0.60**), PL (r = 0.77**), FW (r = 0.78**) and DW (r = 0.85**) (vegetative traits). The implications of these results for plant breeding and forage management purposes are discussed.