Tumour necrosis factor -308 and -238 promoter polymorphisms are predictors of a null virological response in the treatment of Brazilian hepatitis C patients

Certain host single nucleotide polymorphisms (SNPs) affect the likelihood of a sustained virological response (SVR) to treatment in subjects infected with hepatitis C virus (HCV). SNPs in the promoters of interleukin (IL)-10 (-1082 A/G, rs1800896), myxovirus resistance protein 1 (-123 C/A, rs17000900 and -88 G/T, rs2071430) and tumour necrosis factor (TNF) (-308 G/A, rs1800629 and -238 G/A, rs361525) genes and the outcome of PEGylated α-interferon plus ribavirin therapy were investigated. This analysis was performed in 114 Brazilian, HCV genotype 1-infected patients who had a SVR and in 85 non-responders and 64 relapsers. A significantly increased risk of having a null virological response was observed in patients carrying at least one A allele at positions -308 [odds ratios (OR) = 2.58, 95% confidence intervals (CI) = 1.44-4.63, p = 0.001] or -238 (OR = 7.33, 95% CI = 3.59-14.93, p < 0.001) in the TNF promoter. The risk of relapsing was also elevated (-308: OR = 2.87, 95% CI = 1.51-5.44, p = 0.001; -238: OR = 4.20, 95% CI = 1.93-9.10, p < 0.001). Multiple logistic regression of TNF diplotypes showed that patients with at least two copies of the A allele had an even higher risk of having a null virological response (OR = 16.43, 95% CI = 5.70-47.34, p < 0.001) or relapsing (OR = 6.71, 95% CI = 2.18-20.66, p = 0.001). No statistically significant association was found between the other SNPs under study and anti-HCV therapy response.

Diagnostic challenges of single plaque-like lesion paucibacillary leprosy

The diagnosis of single-lesion paucibacillary leprosy remains a challenge. Reviews by expert dermatopathologists and quantitative polymerase chain reaction (qPCR) results obtained from 66 single-plaque biopsy samples were compared. Histological findings were graded as high (HP), medium (MP) or low (LP) probability of leprosy or other dermatopathy (OD). Mycobacterium leprae-specific genes were detected using qPCR. The biopsies of 47 out of 57 clinically diagnosed patients who received multidrug therapy were classified as HP/MP, eight of which were qPCR negative. In the LP/OD (n = 19), two out of eight untreated patients showed positive qPCR results. In the absence of typical histopathological features, qPCR may be utilised to aid in final patient diagnosis, thus reducing overtreatment and delay in diagnosis.

Using a single tablet daily to treat latent tuberculosis infection in Brazil: bioequivalence of two different isoniazid formulations (300 mg and 100 mg) demonstrated by a sensitive and rapid high-performance liquid chromatography-tandem mass spectrometry method in a randomised, crossover study

The recommended treatment for latent tuberculosis (TB) infection in adults is a daily dose of isoniazid (INH) 300 mg for six months. In Brazil, INH was formulated as 100 mg tablets. The treatment duration and the high pill burden compromised patient adherence to the treatment. The Brazilian National Programme for Tuberculosis requested a new 300 mg INH formulation. The aim of our study was to compare the bioavailability of the new INH 300 mg formulation and three 100 mg tablets of the reference formulation. We conducted a randomised, single dose, open label, two-phase crossover bioequivalence study in 28 healthy human volunteers. The 90% confidence interval for the INH maximum concentration of drug observed in plasma and area under the plasma concentration vs. time curve from time zero to the last measurable concentration “time t” was 89.61-115.92 and 94.82-119.44, respectively. The main limitation of our study was that neither adherence nor the safety profile of multiple doses was evaluated. To determine the level of INH in human plasma, we developed and validated a sensitive, simple and rapid high-performance liquid chromatography-tandem mass spectrometry method. Our results showed that the new formulation was bioequivalent to the 100 mg reference product. This finding supports the use of a single 300 mg tablet daily strategy to treat latent TB. This new formulation may increase patients’ adherence to the treatment and quality of life.

A molecular platform for the diagnosis of multidrug-resistant and pre-extensively drug-resistant tuberculosis based on single nucleotide polymorphism mutations present in Colombian isolates of Mycobacterium tuberculosis

Developing a fast, inexpensive, and specific test that reflects the mutations present in Mycobacterium tuberculosis isolates according to geographic region is the main challenge for drug-resistant tuberculosis (TB) control. The objective of this study was to develop a molecular platform to make a rapid diagnosis of multidrug-resistant (MDR) and extensively drug-resistant TB based on single nucleotide polymorphism (SNP) mutations present in therpoB, katG, inhA,ahpC, and gyrA genes from Colombian M. tuberculosis isolates. The amplification and sequencing of each target gene was performed. Capture oligonucleotides, which were tested before being used with isolates to assess the performance, were designed for wild type and mutated codons, and the platform was standardised based on the reverse hybridisation principle. This method was tested on DNA samples extracted from clinical isolates from 160 Colombian patients who were previously phenotypically and genotypically characterised as having susceptible or MDR M. tuberculosis. For our method, the kappa index of the sequencing results was 0,966, 0,825, 0,766, 0,740, and 0,625 forrpoB, katG, inhA,ahpC, and gyrA, respectively. Sensitivity and specificity were ranked between 90-100% compared with those of phenotypic drug susceptibility testing. Our assay helps to pave the way for implementation locally and for specifically adapted methods that can simultaneously detect drug resistance mutations to first and second-line drugs within a few hours.

Exposure of a single host (Chrysomya megacephala) (Calliphoridae) to different quantities of female parasitoids (Nasonia vitripennis) (Pteromalidae)

The aim of this study was to verify the duration of the development period, number of parasitoids produced per pupa, parasitism rate and sex ratio of Nasonia vitripennis (Hymenoptera, Pteromalidae), when they were exposed to a single host: Chrysomya megacephala (Diptera, Calliphoridae). One pupa was exposed in glass tubes to different numbers of female parasitoids (1, 3, 5, 7, 9 and 11) during 48 h. Twenty replications/treatment were used, under controlled conditions (T= 27 °C day/ 25 °C night, 60 ± 10% RH). Statistical analysis of the data was made using the ANOVA test and the "a posteriori" comparisons were made using the Tukey-HSD test (both tests with a significance level of 5%). The duration of the development period was longer in treatments where a higher density of females per host was used. When five females per host were used, the mean number of parasitoids that emerged per pupa was higher. The data showed a tendency to a decrease in the amount of parasitoids emerged per host, especially of female, when used high quantities of female per host. Higher parasitism rates were observed in the 3:1 and 5:1 treatments and an increase in the percentage of unviable pupae was observed, probably due to an increase of female densities in the treatments, possibly a consequence of superparasitism.

Infestation and natural parasitism of aphids in single and mixed pastures of black oats and ryegrass

Some species of aphids are major pests on cereal crops and grass pastures. Usually these pests are not adequately controlled in pasture lands that become sources of aphid infestations to cereal crops. The dynamics of aphids and the incidence of natural enemies are less known in pasture systems than in cereal fields. The objective of this work was to assess the aphid infestation and natural aphid parasitism in different pasture composition. Three hypotheses were tested: 1- the aphid species composition in pastures may vary according to the cereal species in the field; 2- the mixture of two plant species can modify the amount and diversity of aphids; 3- the plant species composition of pasture fields influences the parasitism of aphids. Empirical data were obtained from three Poaceae fields: black oats (Avena strigosa L.), ryegrass (Lolium multiflorum L.), and a mixed field of black oats and ryegrass. The most abundant aphid species was Rhopalosiphum padi followed by Sitobion avenae. Plant species composition increases the amount and the parasitism rates of aphids. The mixture of heavily infested black oats with a poorly infested ryegrass resulted in reduction of aphid infestation in comparison with heavily-infested single plant species field. This is possible because the conditions are favorable for the development of parasitoid populations. Aphidius colemani was the main parasitoid found in these areas.

Método convencional e "bootstrap" para estimar o número de observações na determinação dos parâmetros da função K(Θ)

A variabilidade natural do solo dificulta a obtenção de valores que representem adequadamente as propriedades do solo em determinada área. O conhecimento do número mínimo de observações, que devem ser realizadas para representar, com um erro aceitável, uma propriedade ou característica do solo, é fundamental para que os resultados experimentais possam ser aplicados com segurança. No presente trabalho, comparou-se o método convencional (teste-t) com o método "bootstrap", com vistas em estimar o número de observações necessárias para calcular os parâmetros que caracterizam a relação entre a condutividade hidráulica e o teor de água do solo, determinada pelo método do perfil instantâneo. Realizou-se um experimento de drenagem num Latossolo Vermelho-Amarelo em Piracicaba (SP), numa parcela experimental com 45 pontos de observação distanciados de 1 m entre si. Observaram-se a umidade (com TDR) e o potencial mátrico (com tensiômetros) durante três semanas de redistribuição da água. Após processamento dos dados, o conjunto de valores mostrou uma distribuição não-normal, fazendo-se necessária a eliminação de "outliers" para a aplicação do método convencional, normalizando a distribuição. Assim, o uso do método tradicional só é recomendado após a confirmação da pertinência da eliminação dos "outliers". Ambos os métodos de análise requerem grande número de repetições, reafirmando que determinações da função condutividade hidráulica com poucas repetições não podem ser extrapoladas para áreas maiores.

Influence of organic-mineral fertilization of an oxisol on soil chemical properties and Bracharia brizantha production

The use of organic-mineral fertilizer produced by the manufacturing industry of lysine and threonine amino acids can improve the fertility of tropical soils. The objective of this study was to evaluate the influence of different doses of the organic-mineral fertilizer named Ajifer L-14 on chemical properties and on the response with increased production of a forage on a Red Latosol in the northwestern region of São Paulo State, Brazil. A randomized block design was used with seven treatments and four replications. The treatments consisted of: T1- control (without application of Ajifer L-14); T2- control (natural vegetation); T3- mineral fertilization according to crop requirements and soil analysis (application of 1.35 kg plot-1 of urea, 2.20 single superphosphate, and 0.51 KCl, corresponding to 60 of N, 40 P2O5 and 30 kg ha-1 of K2O); T4- fertilization with Ajifer L-14 according to the recommendation resulting from the soil chemical analysis (40 L plot-1, corresponding to 60 kg ha-1 N); T5- fertilization with Ajifer L-14, at a rate of 150 % of the recommended values (60 L plot-1, corresponding to 90 kg ha-1 N); T6- fertilization with Ajifer L-14 at a rate of 50 % of the recommended values (20 L plot-1, corresponding to 30 kg ha-1 N); T7- fertilization with Ajifer L-14 at a rate of 125 % of the recommended values (50 L plot-1, corresponding to 75 kg ha-1 N); T8- fertilization with Ajifer L-14 at a rate of 75 % of the recommended values (30 L plot-1, corresponding to 45 kg ha-1 N). The following soil chemical properties were evaluated (layers 0.0-0.1 and 0.1-0.2 m): P, organic matter, pH, K+, Ca2+, Mg2+, cation exchange capacity, potential acidity, and base saturation. The application of this organic-mineral fertilizer does not influence the soil chemical properties. Regression analysis indicated a polynomial relationship between the application rates of organic-mineral fertilizer and the production of dry matter and crude protein of Bracharia Brizantha.

Proposal for an index to classify irrigation water quality: a case study in northeastern Brazil

One way of classifying water quality is by means of indices, in which a series of parameters analyzed are joined a single value, facilitating the interpretation of extensive lists of variables or indicators, underlying the classification of water quality. The objective of this study was to develop a statistically based index to classify water according to the Irrigation Water Quality Index (IWQI), to evaluate the ionic composition of water for use in irrigation and classify it by its source. For this purpose, the database generated during the Technology Generation and Adaptation (GAT) program was used, in which, as of 1988, water samples were collected monthly from water sources in the states of Paraíba, Rio Grande do Norte and Ceará. To evaluate water quality, the electrical conductivity (EC) of irrigation water was taken as a reference, with values corresponding to 0.7 dS m-1. The chemical variables used in this study were: pH, EC, Ca, Mg, Na, K, Cl, HCO3, CO3, and SO4. The data of all characteristics evaluated were standardized and data normality was confirmed by Lilliefors test. Then the irrigation water quality index was determined by an equation that relates the standardized value of the variable with the number of characteristics evaluated. Thus, the IWQI was classified based on indices, considering normal distribution. Finally, these indices were subjected to regression analysis. The method proposed for the IWQI allowed a satisfactory classification of the irrigation water quality, being able to estimate it as a function of EC for the three water sources. Variation in the ionic composition was observed among the three sources and within a single source. Although the water quality differed, it was good in most cases, with the classification IWQI II.

Soil thermal diffusivity estimated from data of soil temperature and single soil component properties

Under field conditions, thermal diffusivity can be estimated from soil temperature data but also from the properties of soil components together with their spatial organization. We aimed to determine soil thermal diffusivity from half-hourly temperature measurements in a Rhodic Kanhapludalf, using three calculation procedures (the amplitude ratio, phase lag and Seemann procedures), as well as from soil component properties, for a comparison of procedures and methods. To determine thermal conductivity for short wave periods (one day), the phase lag method was more reliable than the amplitude ratio or the Seemann method, especially in deeper layers, where temperature variations are small. The phase lag method resulted in coherent values of thermal diffusivity. The method using properties of single soil components with the values of thermal conductivity for sandstone and kaolinite resulted in thermal diffusivity values of the same order. In the observed water content range (0.26-0.34 m³ m-3), the average thermal diffusivity was 0.034 m² d-1 in the top layer (0.05-0.15 m) and 0.027 m² d-1 in the subsurface layer (0.15-0.30 m).

Can Soil Penetration Resistance and Bulk Density Be Determined in a Single Undisturbed Sample?

Soil quality indicators such as penetration resistance (PR) and bulk density (BD) are traditionally determined in a single undisturbed soil sample. The aim of this study was to assess the effect of PR measurements of undisturbed samples on the determination of BD in the same sample of two soils differing in clay contents. To this end, samples were collected from the 0.00-0.10 and 0.10-0.20 m layers of two soils of clayey and very clayey texture. Volumetric rings were used to collect a total of 120 undisturbed soil samples from each soil layer that were divided into two subsets containing 60 units each. One sample set, designated “perforated samples”, was used to determine PR and BD in the same undisturbed sample; the other, named “intact samples”, was used to determine BD only. Bulk density values for perforated and intact samples were compared by analysis of variance, using a completely randomized experimental design. Means were compared by the t-test at 5 %. The BD values for the clayey soil were similar in perforated and intact samples from the two layers. However, BD of the very clayey soil was lower in the perforated than in the intact samples at both depths. Therefore, PR and BD in clayey soils can be accurately determined in the same undisturbed sample whereas in very clayey soils, different samples are required for this purpose.