The fluorolysis assay, a highly sensitive method for measuring the cytolytic activity of T cells at very low numbers

We have developed a highly sensitive cytolysis test, the fluorolysis assay, as a simple nonradioactive and inexpensive alternative to the standard Cr-51-release assay. P815 cells were stably transfected with a plasmid expressing the enhanced green fluorescent protein (EGFP) gene. These target cells were coated with or without cognate peptide or anti-CD3 Ab and then incubated with CD8(+) T cells to allow antigen-specific or nonspecific lysis. The degree of target cell lysis was measured using flow cytometry to count the percentage of viable propidium iodide(-) EGFP(+) cells, whose numbers were standardized to a reference number of fluorochrome-linked beads. By using small numbers of target cells (200-800 per reaction) and extended incubation times (up to 2 days), the antigen-specific cytolytic activity of one to two activated CD8(+) T cells of a CTL line could be detected. The redirected fluorolysis assay also measured the activity of very few ( greater than or equal to6) primary CD8(+) T cells following polyclonal activation. Importantly, antigen-specific lysis by small numbers ( greater than or equal to 25) of primary CD8(+) T cells could be directly measured ex vivo. This exquisite sensitivity of the fluorolysis assay, which was at least 8-33-folds higher than an optimized 51 Cr-release assay, allows in vitro and ex vivo studies of immune responses that would otherwise not be possible due to low CTL numbers or frequencies. (C) 2002 Elsevier Science B.V. All rights reserved.

Comparison of an ELISA and an LC/MS/MS method for measuring tacrolimus concentrations and making dosage decisions in transplant recipients

This study compared an enzyme-linked immunosorbent assay (ELISA) to a liquid chromatography-tandem mass spectrometry (LC/MS/MS) technique for measurement of tacrolimus concentrations in adult kidney and liver transplant recipients, and investigated how assay choice influenced pharmacokinetic parameter estimates and drug dosage decisions. Tacrolimus concentrations measured by both ELISA and LC/MS/MS from 29 kidney (n = 98 samples) and 27 liver (n = 97 samples) transplant recipients were used to evaluate the performance of these methods in the clinical setting. Tacrolimus concentrations measured by the two techniques were compared via regression analysis. Population pharmacokinetic models were developed independently using ELISA and LC/MS/MS data from 76 kidney recipients. Derived kinetic parameters were used to formulate typical dosing regimens for concentration targeting. Dosage recommendations for the two assays were compared. The relation between LC/MS/MS and ELISA measurements was best described by the regression equation ELISA = 1.02 . (LC/MS/MS) + 0.14 in kidney recipients, and ELISA = 1.12 . (LC/MS/MS) - 0.87 in liver recipients. ELISA displayed less accuracy than LC/MS/MS at lower tacrolimus concentrations. Population pharmacokinetic models based on ELISA and LC/MS/MS data were similar with residual random errors of 4.1 ng/mL and 3.7 ng/mL, respectively. Assay choice gave rise to dosage prediction differences ranging from 0% to 30%. ELISA measurements of tacrolimus are not automatically interchangeable with LC/MS/MS values. Assay differences were greatest in adult liver recipients, probably reflecting periods of liver dysfunction and impaired biliary secretion of metabolites. While the majority of data collected in this study suggested assay differences in adult kidney recipients were minimal, findings of ELISA dosage underpredictions of up to 25% in the long term must be investigated further.

Measuring range of active cervical rotation in a position of full head flexion using the 3D Fastrak measurement system: an intra-tester reliability study

Most external assessments of cervical range of motion assess the upper and lower cervical regions simultaneously. This study investigated the within and between days reliability of the clinical method used to bias this movement to the upper cervical region, namely measuring rotation of the head and neck in a position of full cervical flexion. Measurements were made using the Fastrak measurement system and were conducted by one operator. Results indicated high levels of within and between days repeatability (range of ICC2,1 values: 0.85-0.95). The ranges of axial rotation to right and left, measured with the neck positioned in full flexion, were approximately 56% and 50%, respectively of total cervical rotation, which relates well to the proportional division of rotation in the upper and lower cervical regions. These results suggest that this method of measuring rotation would be appropriate for use in subject studies where movement dysfunction is present in the upper cervical region, such as those with cervicogenic headache. (C) 2003 Elsevier Science Ltd. All rights reserved.

On measuring quality of a probabilistic commodity forecast for a system that incorporates seasonal climate forecasts

Regional commodity forecasts are being used increasingly in agricultural industries to enhance their risk management and decision-making processes. These commodity forecasts are probabilistic in nature and are often integrated with a seasonal climate forecast system. The climate forecast system is based on a subset of analogue years drawn from the full climatological distribution. In this study we sought to measure forecast quality for such an integrated system. We investigated the quality of a commodity (i.e. wheat and sugar) forecast based on a subset of analogue years in relation to a standard reference forecast based on the full climatological set. We derived three key dimensions of forecast quality for such probabilistic forecasts: reliability, distribution shift, and change in dispersion. A measure of reliability was required to ensure no bias in the forecast distribution. This was assessed via the slope of the reliability plot, which was derived from examination of probability levels of forecasts and associated frequencies of realizations. The other two dimensions related to changes in features of the forecast distribution relative to the reference distribution. The relationship of 13 published accuracy/skill measures to these dimensions of forecast quality was assessed using principal component analysis in case studies of commodity forecasting using seasonal climate forecasting for the wheat and sugar industries in Australia. There were two orthogonal dimensions of forecast quality: one associated with distribution shift relative to the reference distribution and the other associated with relative distribution dispersion. Although the conventional quality measures aligned with these dimensions, none measured both adequately. We conclude that a multi-dimensional approach to assessment of forecast quality is required and that simple measures of reliability, distribution shift, and change in dispersion provide a means for such assessment. The analysis presented was also relevant to measuring quality of probabilistic seasonal climate forecasting systems. The importance of retaining a focus on the probabilistic nature of the forecast and avoiding simplifying, but erroneous, distortions was discussed in relation to applying this new forecast quality assessment paradigm to seasonal climate forecasts. Copyright (K) 2003 Royal Meteorological Society.

Measuring the accumulated hazards of smoking: global and regional estimates for 2000

Objective: Current prevalence of smoking, even where data are available, is a poor proxy for cumulative hazards of smoking, which depend on several factors including the age at which smoking began, duration of smoking, number of cigarettes smoked per day, degree of inhalation, and cigarette characteristics such as tar and nicotine content or filter type. Methods: We extended the Peto-Lopez smoking impact ratio method to estimate accumulated hazards of smoking for different regions of the world. Lung cancer mortality data were obtained from the Global Burden of Disease mortality database. The American Cancer Society Cancer Prevention Study, phase 11 (CPS-II) with follow up for the years 1982 to 1988 was the reference population. For the global application of the method, never-smoker lung cancer mortality rates were chosen based on the estimated use of coal for household energy in each region. Results: Men in industrialised countries of Europe, North America, and the Western Pacific had the largest accumulated hazards of smoking. Young and middle age males in many regions of the developing world also had large smoking risks. The accumulated hazards of smoking for women were highest in North America followed by Europe. Conclusions: In the absence of detailed data on smoking prevalence and history, lung cancer mortality provides a robust indicator of the accumulated hazards of smoking. These hazards in developing countries are currently more concentrated among young and middle aged males.

Sistema de Controle log??stico de Medicamentos Antirretrovirais (SICLOM)

O Sistema de Controle Log??stico de Medicamentos Antirretrovirais (Siclom) ?? uma ferramenta criada com o objetivo de gerenciamento log??stico dos medicamentos antirretrovirais (ARV) e de aprimoramento da qualifica????o da dispensa????o. Esse sistema permite que o departamento de DST, Aids e Hepatites Virais, secretarias estaduais e municipais de Sa??de mantenham-se atualizados em rela????o ao fornecimento, dispensa????o de medicamentos aos pacientes em tratamento conforme as recomenda????es existentes no Consenso Terap??utico Brasileiro, al??m do controle dos estoques de cada medicamento nas v??rias regi??es do pa??s. As informa????es de consumo e estoques s??o fundamentais para que o adequado suprimento aos estados ocorra evitando, assim, a ruptura dos estoques locais e preju??zo no atendimento aos pacientes. Atualmente utilizam o sistema 670 unidades dispensadoras de medicamentos, distribu??das em todo o Brasil; 27 coordena????es estaduais de DST/Aids; 61 almoxarifados; e 936 maternidades

Log??stica de distribui????o e entrega dos objetos do Programa Nacional do Livro Did??tico (PNLD)

O Programa Nacional do Livro Did??tico ??? PNLD, administrado pelo Fundo Nacional de Desenvolvimento da Educa????o ??? FNDE, utiliza, desde 1994, os servi??os da Empresa Brasileira de Correios e Tel??grafos - ECT, para levar ??s escolas p??blicas mais de uma centena de milh??es de livros did??ticos. V??rios eram os problemas a serem resolvidos. Antes de a ECT assumir os servi??os, somente 25% dos livros eram entregues antes do in??cio do ano letivo e o restante entregue durante ano, prejudicando de uma sobre maneira, n??o s?? alunos e professores, mas todo o planejamento de ensino. A aus??ncia de um controle efetivo da entrega dos livros, aliado ao endere??amento incorreto, gerava um de alto ??ndice de reclama????o quanto ao n??o recebimento dos objetos. Com as a????es descritas nesse relato, foi poss??vel aumentar o n??mero de crian??as/adolescentes estudantes de escolas p??blicas, com cobertura de todos os munic??pios brasileiros, os quais s??o na realidade os grandes clientes de todo o processo. Certamente que o sucesso da opera????o envolve a FNDE e a ECT, sendo que nesta ??ltima ?? imprescind??vel a perfeita sintonia entre todas as ??reas

Cataloga????o, uma ferramenta para a log??stica

A cataloga????o consiste numa ferramenta de aplica????o nos ??rg??os p??blicos federais, que permite o uso de linguagem ??nica de identifica????o de materiais, de forma que possa ser entendida e manipulada por qualquer sistema, desde que este conhe??a as regras da linguagem. Essa ferramenta, entre outras utilidades, possibilita a execu????o de boa gest??o de material, por meio da intera????o de todos os ??rg??os do governo, permitindo o apoio m??tuo entre eles, e da nacionaliza????o de itens importados, contribuindo para a redu????o de nossa depend??ncia externa e para o incentivo ?? ind??stria nacional. A cataloga????o possibilita, ainda, a descoberta de utiliza????o de itens comuns de material, a perfeita identifica????o de um item, a otimiza????o do planejamento or??ament??rio e a redu????o do custo log??stico dos ??rg??os por melhor compra, armazenagem, aplica????o, destina????o de excessos e otimiza????o de estoques

Sistema de avalia????o log??stica variada

As normas que regem a gratifica????o de Desempenho e Produtividade ??? GDP ??? recebidas pelos servidores da Carreira de Finan??as e Controle da Secretaria Federal de Controle, estabelecem limites pares ao n??mero de servidores que podem ser avaliados com notas superiores a 8. Essa limita????o est?? fixada em 20% para as notas na faixa de 8 a 9 e 10% para as notas acima de 9. O fato da Secretaria Federal de Controle possuir muitas unidades, inviabilizou a escolha dos servidores que, devido ao seu melhor desempenho, mereceriam receber as notas mais altas. A solu????o deste e de outros problemas dele decorrentes foi a implementa????o do Sistema de Avalia????o Log??stica Variada ??? SALVA, que operacionaliza a distribui????o dos valores n??o inteiros da gratifica????o de desempenho e produtividade com exatid??o e igualdade

M??dulo 1 - a import??ncia e a necessidade da lei de licita????es: curso legisla????o aplicada ?? log??stica de suprimentos lei 8.666/93, preg??o e registro de pre??os

Este documento trata de: aspectos conceituais da lei: finalidade, import??ncia e hierarquia da lei.; no????es gerais da lei de licita????es - Lei n?? 8.666/93; tipos de licita????o:menor pre??o, melhor t??cnica, t??cnica e pre??o e maior lance ou oferta; modalidades de licita????o: concorr??ncia, tomada de pre??os, convite, concurso e leil??o; exce????es ?? obrigatoriedade de licita????o: dispensa e inexigibilidade; regime de execu????o indireta; comiss??o de licita????o; etapas do processo licitat??rio: edital, procedimentos/documentos do certame, registro cadastral, habilita????o dos interessados, julgamento e encerramento; preg??o; registro de pre??os