Association of hepatitis C virus NS5B variants with resistance to new antiviral drugs among untreated patients

Mutations located in the 109-amino acid fragment of NS5B are typically associated with resistance to interferon (IFN) and ribavirin (RIB) and to new antiviral drugs. The prevalence of these mutations was examined in 69 drug-naïve individuals with hepatitis C virus (HCV) infections in Rio de Janeiro, Brazil. Mutations related to non-response to IFN/RIB were observed in all subtypes studied (1a, 1b, 2b, 3a and 4). The most common mutation was Q309R, present in all subtypes, except subtype 2b with frequency above 20%. D244N was detected only in subtype 3a and A333E was detected only in subtype 2b. We did not detect the S282T, S326G or T329I mutations in any of the samples analysed. Of note, the C316N mutation, previously related to a new non-nucleoside compound (HCV796 and AG-021541), was observed in only eight of 33 (24%) samples from subtype 1b. Site 316 was under positive selection in this HCV variant. Our data highlight the presence of previously described resistance mutations in HCV genotypes from drug-naïve patients.

The first report of the qnrB19, qnrS1 and aac(6´)-Ib-cr genes in urinary isolates of ciprofloxacin-resistant Escherichia coli in Brazil

In this study, we investigated the presence of plasmid-mediated quinolone resistance (PMQR) genes among 101 ciprofloxacin-resistant urinary Escherichia coli isolates and searched for mutations in the quinolone-resistance-determining regions (QRDRs) of the DNA gyrase and topoisomerase IV genes in PMQR-carrying isolates. Eight isolates harboured the qnr and aac(6')-Ib-cr genes (3 qnrS1, 1 qnrB19 and 4 aac(6')-Ib-cr). A mutational analysis of the QRDRs in qnr and aac(6')-Ib-cr-positive isolates revealed mutations in gyrA, parC and parE that might be associated with high levels of resistance to quinolones. No mutation was detected in gyrB. Rare gyrA, parC and parE mutations were detected outside of the QRDRs. This is the first report of qnrB19, qnrS1 and aac(6')-Ib-cr -carrying E. coli isolates in Brazil.

Genetic diversity and antimicrobial resistance in Streptococcus agalactiae strains recovered from female carriers in the Bucharest area

For the first time, we used multilocus sequence typing (MLST) to understand how Romanian group B streptococcus (GBS) strains fit into the global GBS population structure. Colonising isolates recovered from adult human females were tested for antibiotic resistance, were molecularly serotyped based on the capsular polysaccharide synthesis (cps) gene cluster and further characterised using a set of molecular markers (surface protein genes, pilus-encoded islands and mobile genetic elements inserted in the scpB-lmb intergenic region). Pulsed-field gel electrophoresis was used to complement the MLST clonal distribution pattern of selected strains. Among the 55 strains assigned to six cps types (Ia, Ib, II-V), 18 sequence types (STs) were identified by MLST. Five STs represented new entries to the MLST database. The prevalent STs were ST-1, ST-17, ST-19 and ST-28. Twenty molecular marker profiles were identified. The most common profiles (rib+GBSi1+PI-1, rib+GBSi1+PI-1, PI-2b and alp2/3+PI-1, PI-2a) were associated with the cps III/ST-17 and cps V/ST-1 strains. A cluster of fluoroquinolone-resistant strains was detected among the cps V/ST-19 members; these strains shared alp1 and IS1548 and carried PI-1, PI-2a or both. Our results support the usefulness of implementing an integrated genotyping system at the reference laboratory level to obtain the reliable data required to make comparisons between countries.

Genetic associations in the detection of QTLs for wheat spike-related traits

The objective of this work was to assess the genetic diversity and population structure of wheat genotypes, to detect significant and stable genetic associations, as well as to evaluate the efficiency of statistical models to identify chromosome regions responsible for the expression of spike-related traits. Eight important spike characteristics were measured during five growing seasons in Serbia. A set of 30 microsatellite markers positioned near important agronomic loci was used to evaluate genetic diversity, resulting in a total of 349 alleles. The marker-trait associations were analyzed using the general linear and mixed linear models. The results obtained for number of allelic variants per locus (11.5), average polymorphic information content value (0.68), and average gene diversity (0.722) showed that the exceptional level of polymorphism in the genotypes is the main requirement for association studies. The population structure estimated by model-based clustering distributed the genotypes into six subpopulations according to log probability of data. Significant and stable associations were detected on chromosomes 1B, 2A, 2B, 2D, and 6D, which explained from 4.7 to 40.7% of total phenotypic variations. The general linear model identified a significantly larger number of marker-trait associations (192) than the mixed linear model (76). The mixed linear model identified nine markers associated to six traits.

Um panorama atual da química e da farmacologia de naftoquinonas, com ênfase na beta-lapachona e derivados

Naphthoquinones have been extensively studied due to their activity as topoisomerase inhibitors. These enzymes are critical to DNA replication in cells. In addition, naphthoquinones have been shown to induce what are known as "reactive oxygen species" that can cause damage to cells. beta-Lapachone is a very important pyranaphthoquinone obtained from the heartwood of the lapacho tree, Tabebuia avellanedae Lorentz ex. Griseb. (Bignoniaceae), and other Tabebuia trees native to Central and South America and chemically from lapachol. beta-Lapachone has a diversity of useful biological activities against various cancer cell lines such as human ovarian and prostate tumors and, at lower doses is a radiosensitizer of several human cancer cell lines. It gives rise to a variety of effects in vitro including the inhibition or activation of topoisomerase I an II in a distinct manner from that of other topoisomerase inhibitors. This review intend to discuss some details of the mechanisms of quinone-induced cell damage and death, and we also summarize results of the literature indicating that b-Lapachone may take part in quinone-elicited apoptosis despite the fact that its mechanism of action in vivo and its targets are still unknown.

Síntese, caracterização e estudos de interação de um análogo da antitoxina CcdA empregando fluorescência no estado estacionário

Toxin-antitoxin (TA) systems contribute to plasmid stability by a mechanism called post-segregational killing. The ccd was the first TA system to be discovered with CcdB being the toxin and CcdA the antitoxin. CcdA, an 8.3 kDa protein, interacts with CcdB (11.7 kDa), preventing the cytotoxic activity of CcdB on the DNA gyrase. As an approach to understanding this interaction, CcdA41, a polypeptide derived from CcdA, was synthesized by solid-phase methodology and its interaction with CcdB was analyzed by steady state fluorescence. CcdA41 formed a stable complex with CcdBET2, a peptide based on CcdB, the more recently described bacterial topoisomerase inhibitor.

Validation of a spectrophotometric method to estimate the adsorption on nanoemulsions of an antimalarial oligonucleotide

This study describes the validation of a spectrophotometric method to estimate oligonucleotides association with cationic nanoemulsions. Phosphodiester and phosphorothioate oligonucleotides targeting Plasmodium falciparum topoisomerase II were analyzed at 262 nm. Linear response (r > 0.998) was observed from 0.4 to 1.0 nmol/mL, the relative standard deviation values for the intra- and inter-days precision were lower than 2.6% and the recovery ranged from 98.8 to 103.6% for both oligonucleotides. The association efficiency was estimated based on an ultrafiltration/centrifugation method. Oligonucleotides recovery through 30 kDa-membranes was higher than 92%. The extent of oligonucleotides association (42 to 98%) varied with the composition of nanoemulsions

Aplysfistularine: a novel dibromotyrosine derivative isolated from Aplysina fistularis

The new dibromotyrosine derivative 3,5-dibromo-4-[3'dimethylamonium]propoxyphenyl]-N,N,N-trimethylethanamonium, here referred to as aplysfistularine (1), was isolated from the marine sponge Aplysina fistularis along with 2-(3,5-dibromo-4methoxyphenyl)-N,N,N-trimethylethanamonium (2), aplysterol (3) and 24,28-didehydroaplysterol (4). Their identification was performed by mass spectrometry, infrared, ¹H and 13C NMR, and by comparison with literature data. Compound 2 and the mixture of 3 and 4 were tested in vitro (inhibitory activity) with supercoiled DNA relaxation techniques, and showed inhibitory activity on human DNA topoisomerase II-α. Compound 1 was not tested due to paucity of the material.

Phytochemical investigation of Wissadula periplocifolia (L.) C. Presl and evaluation of its antibacterial activity

A phytochemical study on the aerial parts of Wissadula periplocifolia using chromatographic techniques has led to the isolation of sitosterol (1a), stigmasterol (1b), sitosterol 3-O-β-D-glucopyranoside (2a), stigmasterol 3-O-β-D-glucopyranoside (2b), phaeophytin A (3), 13²-hydroxy-(13²-S)-phaeophytin A (4), phaeophytin B (5), 17³-ethoxyphaeophorbide (6), 3,4-seco-urs-4(23),20(30)-dien-3-oic acid (7), 3-oxo-21β-H-hop-22(29)-ene (8), dammaradienone (9a), and taraxastenone (9b). The isolated compounds were characterised by spectroscopic analysis. A preliminary assay to evaluate the antibacterial activity of W. periplocifolia extracts and fractions showed that the dichloromethane, ethyl acetate, and n-butanol fractions were active against Enterococcus faecalis.

Anacardic acid derivatives from Brazilian propolis and their antibacterial activity

Propolis is a sticky, gummy, resinous substance collected by honeybees (Apis mellifera L.) from various plant sources, which has excellent medicinal properties. This paper describes the isolation and identification of triterpenoids and anacardic acid derivatives from Brazilian propolis and their antibacterial activity. Their structures were elucidated by ¹H and 13C NMR, including uni- and bidimensional techniques; in addition, comparisons were made with data from academic literature. These compounds were identified as: cardanols (1a + 1b), cardols (2a + 2b), monoene anacardic acid (3), a-amirine (4), b-amirine (5), cycloartenol (6), 24-methylene-cycloartenol (7) and lupeol (8). The determination of the position of the double bond after a reaction with Dimethyl disulfide (DMDS) is described for the phenol derivatives. The ethanolic extract was tested in vitro for antimicrobial activity by using the disc diffusion method and it showed significant results against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Shigella spp.

Bound state solutions of schrödinger equation for a more general exponential screened coulomb potential via Nikiforov-Uvarov method

The arbitrary angular momentum solutions of the Schrödinger equation for a diatomic molecule with the general exponential screened coulomb potential of the form V(r) = (- a / r){1+ (1+ b )e-2b } has been presented. The energy eigenvalues and the corresponding eigenfunctions are calculated analytically by the use of Nikiforov-Uvarov (NU) method which is related to the solutions in terms of Jacobi polynomials. The bounded state eigenvalues are calculated numerically for the 1s state of N2 CO and NO

Geostatistics and remote sensing methods in the classification of images of areas cultivated with citrus

This study compares the precision of three image classification methods, two of remote sensing and one of geostatistics applied to areas cultivated with citrus. The 5,296.52ha area of study is located in the city of Araraquara - central region of the state of São Paulo (SP), Brazil. The multispectral image from the CCD/CBERS-2B satellite was acquired in 2009 and processed through the Geographic Information System (GIS) SPRING. Three classification methods were used, one unsupervised (Cluster), and two supervised (Indicator Kriging/IK and Maximum Likelihood/Maxver), in addition to the screen classification taken as field checking.. Reliability of classifications was evaluated by Kappa index. In accordance with the Kappa index, the Indicator kriging method obtained the highest degree of reliability for bands 2 and 4. Moreover the Cluster method applied to band 2 (green) was the best quality classification between all the methods. Indicator Kriging was the classifier that presented the citrus total area closest to the field check estimated by -3.01%, whereas Maxver overestimated the total citrus area by 42.94%.

Avaliação da reinfecção peritoneal após peritonite fecal em ratos

OBJETIVO: Há poucos estudos sobre os efeitos de uma nova infecção após peritonite séptica. Com o objetivo de compreender melhor esta situação, realizou-se o presente experimento tendo como parâmetro o papel do tempo neste fenômeno. MÉTODO: Foram utilizadas 36 ratas Wistar adultas, submetidas a peritonite fecal com injeção intraperitoneal de uma solução de fezes de ratos. Os animais foram divididos em quatro grupos (n = 9): Grupo 1A - controle: injeção intraperitoneal de solução de fezes com uma quantidade sabidamente letal (10 ml/kg); Grupo 1B - reinfecção: injeção intraperitoneal de solução de fezes com uma quantidade sabidamente não letal (2 ml/kg) e, após 30 dias, injeção de solução de fezes (10 ml/kg); Grupo 2A - controle da reinfecção tardia: injeção intraperitoneal de fezes a 10 ml/kg; Grupo 2B - reinfecção tardia: injeção intraperitoneal de fezes a 2ml/kg e, após quatro meses, injeção de 10ml/kg. RESULTADOS: Todos os nove animais do Grupo 1A morreram no período de sete dias após a injeção da solução de fezes. Já no Grupo 1B, pré-infectado, apenas um animal morreu, 24 horas após a injeção da solução de fezes a 10 ml/kg (p < 0,001). Em relação ao Grupo 2, oito dos nove animais de cada subgrupo morreram no período de sete dias. CONCLUSÕES: Uma sepse peritoneal menor por fezes eleva a resistência orgânica a nova contaminação fecal mais intensa que ocorra após um período curto. Contudo, essa defesa não persiste por tempo mais prolongado.

Avaliação da influência do tempo e do diâmetro na obstrução da janela pleuropericárdica

OBJETIVO: Avaliar a influência do diâmetro e do tempo na obstrução da janela pleuro-pericárdica em cães com pericárdio normal. MÉTODO: Trinta e seis cães mestiços foram divididos em seis grupos: 1 a, 1 b,1 c, 2 a, 2 b, 2 c; n=6 por grupo. Nos grupos 1 a , 1 b, 1 c, à janela foi de 2cm de diâmetro e nos grupos 2 a, 2 b, 2 c, foi de 4cm. Os animais foram reoperados respectivamente após 2, 8 e 12 semanas. Na re-operação, avaliou-se o grau de obstrução através de uma escala de aderência pericárdio-epicárdica e a histopatologia das bordas do pericárdio. RESULTADOS: Numa análise global, observou-se 89% de janelas abertas e 11% de janelas totalmente obstruídas. No grupo com 2cm de diâmetro original, na reoperação, encontrou-se um diâmetro maior em 89% dos cães, enquanto que nos cães com janela original de 4cm, isto ocorreu em 61%. Quando comparou-se os resultados nos cães com janelas de diâmetro igual, mas re-operados em diferentes tempos de pós-operatório, não se observou diferença estatisticamente significante. O mesmo ocorreu quando comparou-se os cães com janelas de diâmetro diferente e re-operados em tempos iguais de pós-operatório. O grau de aderência pericárdio-epicárdica, de acordo com a escala de gradação, não foi diferente entre os vários grupos em função do tempo e diâmetro com exceção do grupo com janela de 4cm e re-operado com 8 semanas (Grupo 2b). As alterações histopatológicas não foram estatisticamente significantes entre os grupos. Em nenhum cão observou-se obstrução da janela pleuropericárdica pelo pulmão. CONCLUSÃO: o tempo e o diâmetro da janela pleuropericardica não influenciaram na obstrução da mesma.

Morfologia e resistência cicatriciais da parede abdominal após laparotomias longitudinal e transversal em coelhos

OBJETIVO: Foram estudadas comparativamente as laparotomias longitudinais paramediana e transversal avaliando-se a resistência cicatricial da parede abdominal e seu aspecto histológico, com peritônio suturado ou não. MÉTODO: 30 coelhos foram distribuídos em dois grupos: Grupo 1 (n = 10) - laparotomia longitudinal, Subgrupo 1A (n = 5) sutura das bainhas anterior e posterior do músculo reto abdominal, bem como do peritônio, Subgrupo 1B (n = 5) sutura da bainha anterior do músculo reto abdominal. Grupo 2 (n = 20) - laparotomia transversa, Subgrupo 2A (n = 5) sutura das bainhas anterior e posterior do músculo reto abdominal, bem como do peritônio, Subgrupo 2B (n = 5) sutura apenas da bainha anterior do músculo reto abdominal, Subgrupo 2C (n = 5) sutura, em plano único, do músculo reto abdominal e de sua bainha anterior, Subgrupo 2D (n = 5) síntese, da bainha posterior do músculo reto abdominal junto com o peritônio e, em seguida, sutura do músculo reto abdominal, complementada por sutura da bainha anterior desse músculo. Após 17 dias, foram retirados dois segmentos peritonio-aponeurotico-musculares da cicatriz para avaliação da resistência e de seus aspectos histológicos. RESULTADOS: O valor da resistência para cada um dos grupos avaliados mostrou 1A > 1B, 1A > 2A e 1B > 2B, e 2B > 2C > 2D > 2A (p = 0,014). Deiscência, infecção e aderências foram mais freqüentes no Grupo 2. A histologia mostrou degeneração e necrose muscular, com sua substituição por tecido conjuntivo fibroso maduro cicatricial. CONCLUSÃO: Esses dados indicam que o corte muscular transversal provoca maior enfraquecimento muscular e que o peritônio deixado aberto não altera a resistência cicatricial.