RESUMO
Peripheral arterial disease (PAD) remains underdiagnosed by primary care and cardiovascular physicians. The office-based assessment of PAD is limited by the need for specialized equipment and the time required for performance of the ankle-brachial index (ABI). We explored whether the accuracy of automated ABI measurement by oscillometry compared favorably with the gold-standard method using continuous-wave Doppler ultrasound. Consecutive patients referred to our university hospital noninvasive vascular laboratory for ABI measurement were invited for participation. Of 205 patients, 201 participated, including 55 with PAD. The ABI was measured by automated oscillometry and Doppler ultrasound. The test of trends revealed a correlation coefficient of 0.78 in the left leg and 0.78 in the right leg (P<0.01 for both). The mean ABI difference between methods was 0.04+/-0.01 and 0.06+/-0.01, respectively, in the left and right legs. The differences between the methods followed a normal distribution. Oscillometric determination of the ABI provides an accurate determination of the ABI in an outpatient population. Our findings show automated oscillometry to be a reliable and easier method of ABI measurement, lowering the barrier to incorporation of this diagnostic test into clinical practice.
Assuntos
Tornozelo/irrigação sanguínea , Determinação da Pressão Arterial/métodos , Pressão Sanguínea , Artéria Braquial/fisiopatologia , Oscilometria , Doenças Vasculares Periféricas/diagnóstico , Doenças Vasculares Periféricas/fisiopatologia , Idoso , Tornozelo/diagnóstico por imagem , Automação , Determinação da Pressão Arterial/normas , Artéria Braquial/diagnóstico por imagem , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Oscilometria/métodos , Oscilometria/normas , Pacientes Ambulatoriais , Doenças Vasculares Periféricas/diagnóstico por imagem , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Ultrassonografia DopplerRESUMO
Here we describe lentivirus-infected cell microarrays for the high-throughput screening of gene function in mammalian cells. To create these arrays, we cultured mammalian cells on glass slides 'printed' with lentiviruses pseudotyped as vesicular stomatitis virus glycoprotein, which encode short hairpin RNA or cDNA. Cells that land on the printed 'features' become infected with lentivirus, creating a living array of stably transduced cell clusters within a monolayer of uninfected cells. The small size of the features of the microarrays (300 microm in diameter) allows high-density spotting of lentivirus, permitting thousands of distinct parallel infections on a single glass slide. Because lentiviruses have a wide cellular tropism, including primary cells, lentivirus-infected cell microarrays can be used as a platform for high-throughput screening in a variety of cell types.
Assuntos
Proteínas de Fluorescência Verde/metabolismo , Lentivirus/genética , Análise em Microsséries/métodos , Animais , Linhagem Celular Transformada , Linhagem Celular Tumoral , Células Cultivadas , Células Dendríticas/metabolismo , Fibroblastos/metabolismo , Regulação da Expressão Gênica/genética , Proteínas de Fluorescência Verde/genética , Células HeLa , Humanos , Processamento de Imagem Assistida por Computador , Lamina Tipo A/genética , Lamina Tipo A/metabolismo , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Glicoproteínas de Membrana/genética , Camundongos , Microscopia de Fluorescência , Fosforilação , Regiões Promotoras Genéticas/genética , Proteínas Quinases/genética , Proteínas Quinases/metabolismo , Interferência de RNA , RNA Interferente Pequeno/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Quinases S6 Ribossômicas/metabolismo , Serina-Treonina Quinases TOR , Antígenos Thy-1/genética , Antígenos Thy-1/metabolismo , Transfecção , Proteínas do Envelope Viral/genética , Proteína Vermelha FluorescenteRESUMO
RNA interference (RNAi)-mediated loss-of-function screening in Drosophila melanogaster tissue culture cells is a powerful method for identifying the genes underlying cell biological functions and for annotating the fly genome. Here we describe the development of living-cell microarrays for screening large collections of RNAi-inducing double-stranded RNAs (dsRNAs) in Drosophila cells. The features of the microarrays consist of clusters of cells 200 mum in diameter, each with an RNAi-mediated depletion of a specific gene product. Because of the small size of the features, thousands of distinct dsRNAs can be screened on a single chip. The microarrays are suitable for quantitative and high-content cellular phenotyping and, in combination screens, for the identification of genetic suppressors, enhancers and synthetic lethal interactions. We used a prototype cell microarray with 384 different dsRNAs to identify previously unknown genes that affect cell proliferation and morphology, and, in a combination screen, that regulate dAkt/dPKB phosphorylation in the absence of dPTEN expression.