A novel diagnostic test detects a low frequency of the hemicentin Gln5345Arg variant among Northern Irish age related macular degeneration patients.

Purpose: Age related macular degeneration (AMD) is a common cause of severe vision loss. Identification of genes involved in AMD will facilitate early detection and ultimately help to identify pathways for treatment for this disorder. The A16,263G mutation in the HEMICENTIN-1 gene produces a non-conservative substitution of arginine for glutamine at codon 5345 which has been implicated in familial AMD. The aim of this study is to develop a rapid diagnostic assay for the detection of this mutation and to evaluate its frequency in a sample of AMD patients. Methods: A primer probe set was designed from exon 104 of the HEMICENTIN-1 gene to differentiate between mutant and wild type alleles. A region spanning the mutation was amplified by PCR using a LightCycler (Roche Diagnostic). The mutation was then detected by melt curve analysis of the hybrid formed between the PCR product and a specific fluorescent probe. The frequency of the mutation within the Northern Ireland population was evaluated by assaying 508 affected AMD patients, 25 possibly affected and 163 controls. Results: This assay clearly discriminates between the A16,263G mutant and wild type HEMICENTIN-1 alleles. The wild type sequence has a single base mismatch with the probe which decreases the stability of the hybrid, resulting in a lower TM (TM=51.27 °C) than that observed for the perfectly matched mutant allele (TM=59.9 °C). The mutant allele was detected in only one of the 696 subjects, an affected AMD patient. Conclusions: We describe a rapid assay for the genotyping of the Gln5345Arg mutation using real-time fluorescence PCR to facilitate rapid processing of samples through combined amplification and detection steps. These characteristics are suitable for a clinical setting where high throughput diagnostic procedures are required. The frequency of this mutation within the Northern Ireland population has been estimated at 0.2%, concurring with previous findings that this mutation is a rare variant associated with AMD. A rapid diagnostic assay will facilitate a reliable and convenient evaluation of the frequency of the Gln5345Arg mutation and its association with AMD within other populations.

Cognitivismo y deficiencia mental.

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Leite cru de quatro regiões leiteiras brasileiras: perspectivas de atendimento dos requisitos microbiológicos estabelecidos pela Instrução Normativa 51

Evidências que o leite produzido e consumido no Brasil nem sempre apresenta a qualidade desejada têm gerado a discussão e desenvolvimento de novas políticas de incentivo à produção leiteira, resultando no desenvolvimento do Programa Nacional de Melhoria da Qualidade do Leite. em complementação, em 2002 o Ministério da Agricultura publicou a Instrução Normativa 51 (IN51), com importantes inovações em relação à conservação e transporte do leite cru, além de estabelecimento de um padrão de qualidade para esse tipo de leite (10(6) UFC/mL), a ser implantado em diferentes prazos nas diferentes regiões do país, a partir de 2005. O presente trabalho teve como objetivo verificar se o leite cru produzido em quatro áreas de quatro estados produtores de leite no Brasil estaria, nesse momento, em condições de cumprir o estabelecido na IN 51, especialmente quanto ao atendimento dos padrões microbiológicos previstos. Amostras de leite cru, coletadas em 210 diferentes propriedades nas regiões de Viçosa, MG (47), Pelotas, RS (50), Londrina, PR (63) e Botucatu, SP (50), foram analisadas quanto aos níveis de contaminação por aeróbios mesófilos, utilizando o PetrifilmTM AC. Parcela significativa das amostras (48,6%) apresentaram contagens acima do determinado pela IN51, sendo 21,3% na região de Viçosa (MG), 56,0% na região de Pelotas (RS), 47,6% na região de Londrina (PR) e 68,0% na região de Botucatu (SP). Considerando as diferenças de cada região, foi possível observar a importância da refrigeração na conservação e transporte da produção, bem como da implantação de boas práticas e assistência técnica nas propriedades. Os resultados obtidos permitem concluir que a adequação às normas estabelecidas pela IN51 pode ser mais difícil em algumas regiões do que em outras, sendo fundamental a adoção da refrigeração na conservação e no transporte da produção, e de programas regionais de assistência a produtores leiteiros.

Tm and Tm-Tb-doped germanate glasses for S-band amplifiers

The mechanism involved in the Tm(3+)((3)F(4)) -> Tb(3+)((7)F(0,1,2)) energy transfer as a function of the Tb concentration was investigated in Tm:Tb-doped germanate (GLKZ) glass. The experimental transfer rate was determined from the best fit of the (3)F(4) luminescence decay due to the Tm -> Tb energy transfer using the Burshtein model. The result showed that the 1700 nm emission from (3)F(4) can be completely quenched by 0.8 mol% of Tb(3+). As a consequence, the (7)F(3) state of Tb(3+) interacts with the (3)H(4) upper excited state of TM(3+) slighting decreasing its population. The effective amplification coefficient beta(cm(-1)) that depends on the population density difference Delta n = n((3)H(4))-n((3)F(4)) involved in the optical transition of Tm(3+) (S-band) was calculated by solving the rate equations of the system for continuous pumping with laser at 792 nm, using the Runge-Kutta numerical method including terms of fourth order. The population density inversion An as a function of Tb(3+) concentration was calculated by computational simulation for three pumping intensities, 0.2, 2.2 and 4.4 kWcm(-2). These calculations were performed using the experimental Tm -> Tb transfer rates and the optical constants of the Tm (0.1 mol%) system. It was demonstrated that 0.2 mol% of Tb(3+) propitiates best population density inversion of Tin(3+) maximizing the amplification coefficient of Tm-doped (0.1 mol%) GLKZ glass when operating as laser intensity amplification at 1.47 mu m. (C) 2007 Elsevier B.V. All rights reserved.