A Novel Diagnostic Target in the Hepatitis C Virus Genome

BackgroundDetection and quantification of hepatitis C virus (HCV) RNA is integral to diagnostic and therapeutic regimens. All molecular assays target the viral 5'-noncoding region (59-NCR), and all show genotype-dependent variation of sensitivities and viral load results. Non-western HCV genotypes have been under-represented in evaluation studies. An alternative diagnostic target region within the HCV genome could facilitate a new generation of assays.Methods and FindingsIn this study we determined by de novo sequencing that the 3'-X-tail element, characterized significantly later than the rest of the genome, is highly conserved across genotypes. To prove its clinical utility as a molecular diagnostic target, a prototype qualitative and quantitative test was developed and evaluated multicentrically on a large and complete panel of 725 clinical plasma samples, covering HCV genotypes 1-6, from four continents (Germany, UK, Brazil, South Africa, Singapore). To our knowledge, this is the most diversified and comprehensive panel of clinical and genotype specimens used in HCV nucleic acid testing (NAT) validation to date. The lower limit of detection (LOD) was 18.4 IU/ml (95% confidence interval, 15.3-24.1 IU/ml), suggesting applicability in donor blood screening. The upper LOD exceeded 10(-9) IU/ml, facilitating viral load monitoring within a wide dynamic range. In 598 genotyped samples, quantified by Bayer VERSANT 3.0 branched DNA (bDNA), X-tail-based viral loads were highly concordant with bDNA for all genotypes. Correlation coefficients between bDNA and X-tail NAT, for genotypes 1-6, were: 0.92, 0.85, 0.95, 0.91, 0.95, and 0.96, respectively; X-tail-based viral loads deviated by more than 0.5 log10 from 5'-NCR-based viral loads in only 12% of samples (maximum deviation, 0.85 log10). The successful introduction of X-tail NAT in a Brazilian laboratory confirmed the practical stability and robustness of the X-tail-based protocol. The assay was implemented at low reaction costs (US$8.70 per sample), short turnover times (2.5 h for up to 96 samples), and without technical difficulties.ConclusionThis study indicates a way to fundamentally improve HCV viral load monitoring and infection screening. Our prototype assay can serve as a template for a new generation of viral load assays. Additionally, to our knowledge this study provides the first open protocol to permit industry-grade HCV detection and quantification in resource-limited settings.

Photoluminescence properties of PZT 52/48 synthesized by microwave hydrothermal method using PVA with template

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

Preparation of hierarchically structured porous aluminas by a dual soft template method

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ischemia and reperfusion in skin flaps: effects of mannitol and vitamin C in reducing necrosis area in a rat experimental model

OBJETIVO: Neste trabalho foi padronizado modelo experimental de isquemia e reperfusão em retalho cutâneo em ratos no qual estudou-se possibilidade de uma solução antioxidante, composta por Ringer lactato, vitamina C e manitol de reduzir a área de necrose. MÉTODOS: O modelo consistiu de levantamento de retalho cutâneo axial de 6,0 x 3,0cm, submetido à isquemia de 8 horas e reperfusão de 7 dias. Os animais foram divididos em quatro grupos: grupos S1, S2 (10 animais cada), C e T (14 animais cada). Nos grupos S1 e S2 todos os procedimentos dos demais grupos foram efetuados, exceto a isquemia e reperfusão: S1 recebeu apenas Ringer lactato e S2 a solução antioxidante. Os grupos C e T foram submetidos à isquemia. O grupo C recebeu somente Ringer lactato e o grupo T a solução antioxidante. No 7(0) dia de pós-operatório as áreas de necrose e pele viável do retalho foram delineadas em decalque de acetato, os quais foram por sua vez analisados em sistema computadorizado KS-300 (Carl Zeiss). RESULTADOS: A análise estatística mostrou que não houve diferenças significativas entre o grupo tratado e controle quanto à área de necrose. CONCLUSÃO: Concluiu-se que o modelo experimental é consistente, determinando área de necrose limitada e uniforme nos animais não tratados e que as drogas usadas, nessa posologia e modo de aplicação, não foram efetivas em diminuir a área de necrose no modelo experimental em questão.

Controlling the growth of zirconia needles precursor from a liquid crystal template

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Zirconia foams prepared by integration of the sol-gel method and dual soft template techniques

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Template-based fluoropolymer ferroelectrets with multiple layers of tubular channels

Significant efforts are devoted to developing new ferroelectrets with well-controlled void distributions or uniform voids and with good long-term and thermal stability of the piezoelectricity. Here, we describe the concept, the fabrication, and the most relevant properties of fluoropolymer ferroelectret systems with three separate films of fluoroethylenepropylene (FEP), alternating with two polytetrafluoroethylene (PTFE) templates. The FEP films are selectively fused by means of a lamination process. Two practically identical PTFE templates are used, which have parallel rectangular openings (1.5×30 mm 2) separated by PTFE ridges of 1.5 mm width. After removing the PTFE templates, a three-layer FEP-film sandwich with tubular channels is obtained. We demonstrate that such FEP-film systems exhibit significant and stable piezoelectricity after charging under a high DC voltage. The resulting piezoelectric effect may be further improved by carefully assembling and arranging the PTFE templates during preparation. ©2010 IEEE.

On the application of nanostructured electrodes prepared by Ti/TiO2/WO3 template : A case study of removing toxicity of indigo using visible irradiation

New assays with HepG2 cells indicate that Indigo Carmine (IC), a dye that is widely used as additive in many food and pharmaceutical industries exhibited cytotoxic effects. This work describes the development of a bicomponent nanostructured Ti/TiO2/WO3 electrode prepared by template method and investigates its efficiency in a photoelectrocatalytic method by using visible light irradiation and applied potential of 1V. After 2h of treatment there are reduction of 97% discoloration, 62% of mineralization and formation of three byproducts assigned as: 2-amine-5-sulfo-benzoic acid, 2,3-dioxo-14-indole-5-sulfonic acid, and 2-amino-α-oxo-5-sulfo-benzeneacetic acid were identified by HPLC-MS/MS. But, cytotoxicity was completely removed after 120min of treatment. © 2013 Elsevier Ltd.

LaNiO3 nanotubes produced using a template-assisted method

We have studied the experimental conditions needed to produce LaNiO3 (LNO) nanostructures using a template-assisted method. In this route, a mesoporous anodic aluminum oxide template was filled with a chemical solution that had been prepared with polymeric precursors route. The precursor solutions and synthesized samples were characterized by X-ray diffraction (XRD), thermogravimetric analysis, infrared spectroscopy and high-resolution scanning electron microscopy (HRSEM). The XRD results for the samples that were heat-treated at 700 degrees C revealed that these samples crystallize in a perovskite-like LaNiO3 structure. HRSEM images revealed that the samples prepared with different deposition times (0.5, 1 and 2 h) promoted the formation of LaNiO3 nanotubes with different wall thicknesses.

The putative Leishmania telomerase RNA (leishTER) undergoes trans-splicing and contains a conserved template sequence

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