183 research outputs found
J Vis Exp
No full textBy virtue of advances in next generation sequencing technologies, we have access to new genome sequences almost daily. The tempo of these advances is accelerating, promising greater depth and breadth. In light of these extraordinary advances, the need for fast, parallel methods to define gene function becomes ever more important. Collections of genome-wide deletion mutants in yeasts and E. coli have served as workhorses for functional characterization of gene function, but this approach is not scalable, current gene-deletion approaches require each of the thousands of genes that comprise a genome to be deleted and verified. Only after this work is complete can we pursue high-throughput phenotyping. Over the past decade, our laboratory has refined a portfolio of competitive, miniaturized, high-throughput genome-wide assays that can be performed in parallel. This parallelization is possible because of the inclusion of DNA 'tags', or 'barcodes,' into each mutant, with the barcode serving as a proxy for the mutation and one can measure the barcode abundance to assess mutant fitness. In this study, we seek to fill the gap between DNA sequence and barcoded mutant collections. To accomplish this we introduce a combined transposon disruption-barcoding approach that opens up parallel barcode assays to newly sequenced, but poorly characterized microbes. To illustrate this approach we present a new Candida albicans barcoded disruption collection and describe how both microarray-based and next generation sequencing-based platforms can be used to collect 10,000-1,000,000 gene-gene and drug-gene interactions in a single experiment.HG000205/HG/NHGRI NIH HHS/United StatesMOP-84305/Canadian Institutes of Health Research/CanadaP01 GH000205/GH/CGH CDC HHS/United StatesR01 HG003317/HG/NHGRI NIH HHS/United StatesT32 HG00044/HG/NHGRI NIH HHS/United States2011-08-11T00:00:00Z21860376PMC321112
Evolução química e história de formação estelar no universo local
No full textTese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Físicas e Matemáticas, Programa de Pós-Graduação em Física, Florianópolis, 2010Analisamos as galáxias do Sloan Digital Sky Survey com o nosso código de síntese espectral Starlight. O Starlight acha a combinação de populações estelares simples de diferentes idades e metalicidades que melhor modela o espectro de uma galáxia. Ele permite derivar vários parâmetros associados a uma galáxia, como a massa em estrelas, a história de formação estelar e a evolução química. A partir do espectro residual puramente nebular (subtraindo do espectro observado o modelado), medimos as linhas de emissão, das quais derivamos propriedades do gás dentro das galáxias. Estudamos a evolução das galáxias com formação estelar. Constatamos que as galáxias de maior massa formaram suas estrelas e seus metais mais rapidamente. A evolução da metalicidade das estrelas é estudada diretamente. Calibramos também a taxa de formação estelar atual medida pela síntese com a medida pela luminosidade de Halfa. Derivamos a relação massa estelar-metalicidade estelar (M*-Z*) em diferentes redshifts. Esta é a primeira vez que a relação M*-Z* é calculada para o mesmo conjunto de galáxias. Observamos que a metalicidade estelar observada tem uma evolução compatível com um modelo simples de evolução química de caixa fechada. Para as galáxias classificadas como LINERs, encontramos que a luminosidade observada em Halfa é compatível com o número de fótons ionizantes emitidos pelas populações estelares velhas dessas galáxias. Este resultado implica em uma profunda revisão da taxa de atividade nuclear nas galáxias do Universo local
Hay - Charles A. Hay
No full textA.B.; D.D., 1859; salutatorian; Philomathaean, orator, anniversary, 1839; Linnaean; entered sophomore class, 1836. Prepared at schools in York and under Dr. Morris. Grad. Gettysburg Seminary, 1841; student, Berlin and Halle, German, 1841-43. Born Feb. 11, 1821, York. Parents, John and Eliza. Lutheran clergyman: Middletown, 1844; Hanover, 1848-49; Harrisburg, 1849-65; Christ, Gettysburg, 1866-93. Prof. of Biblical Literature and German, Gettysburg Seminary, 1844-48; prof. of German, Gettysburg College, 1844-48; prof. of Hebrew, Old Testament Theology and German, Gettysburg Sem., 1865-93. Pres., General Syn., 1881; trustee, Gettysburg College, 1852-93; sec. of board, same, 1867-80. author, Life Sketches of Lutheran Ministers in America. Translator: Luther's Commentary on the Sermon on the Mount; Schmid's Dogmatic Theology. Died June 26, 1893. Married May 5, 1845, Sarah Rebecca Barnitz, York. Children: John W. (class of 1868); Charles E. (class of 1871), Edward G. (class of 1875); Mrs. M.L. Heisler; Mrs. J.A. Himes
Influência da vegetação na iluminação natural de interiores: elementos de controle e entorno
No full textDissertação (mestrado) - Universidade Federal de Santa Catarina, Centro Tecnológico, Programa de Pós-Graduação em Arquitetura e Urbanismo, Florianópolis, 2013.Com o avanço da tecnologia vieram os problemas com o meio ambiente, recursos naturais foram esgotados e uma grande quantidade de resíduo tóxico passou a ser descartada na natureza. Sendo assim, atualmente, a sustentabilidade é o novo ideal a ser seguido pela sociedade, que voltou a entender a importância de integrar a vegetação ao meio em que vivemos. No universo da arquitetura, voltaram a ser integrados ao projeto conceitos como a iluminação natural, mas sua incorporação ao projeto de iluminação deve ser realizada de maneira controlada, de forma a proporcionar um conforto ambiental ao usuário. A vegetação na perspectiva do conforto ambiental tem, atualmente, como foco principal, amenizar a incidência de calor no interior de um edifício. No entanto, seu aproveitamento como contribuinte lumínica é inexistente, sabendo-se apenas que ela possui a capacidade de reduzir a incidência solar direta. O objetivo deste estudo é analisar o comportamento da vegetação como elemento de controle e fonte de luz natural para um ambiente. Para isso, foi desenvolvida uma metodologia onde foram realizados estudos de caso analisando a influência de uma massa vegetal, presente na paisagem, na iluminância de um ponto em um ambiente externo e interno, e em seguida foi elaborado um experimento em um modelo físico em escala reduzida, onde se verificou a iluminância interna com três tipologias de fachadas distintas (um modelo sem nenhuma proteção solar, um modelo com uma obstrução bloqueadora na cor preta e um modelo com obstrução bloqueadora vegetal). Analisando os resultados obtidos, através dos estudos de caso realizados, pôde-se observar que a vegetação possui características importantes que devem ser consideradas durante a elaboração de um projeto lumínico que ter como um elemento de controle uma massa vegetal. Isto porque ela é capaz de refletir, transmitir e difundir a luz natural, ao mesmo tempo em que filtra os raios solares. Abstract : With the advancement of technology came the problems with the natural environment, natural resources have been exhausted and a large amount of toxic residue has to be disposed of in nature. Therefore, sustainability is the new ideal to be followed for the society, which returned to understand the importance of integrating the vegetation in the urban environment. In the world of architecture, return to being integrated the design concepts such as natural lighting. But their incorporation into the lighting design should be performed in a controlled way, in order to provide comfort for the environment user. The vegetation in the perspective of environmental comfort has as its main focus, to reduce the incidence of heat. However, their use as a luminous contributor is nonexistent, the only knowledge is that she has the ability to reduce direct sunlight. The aim of this study is to analyze the behavior of vegetation as an element of control and source of natural light to environment. For this, a methodology was developed where case studies were performed analyzing the influence of a plant mass, present in the landscape, in the illuminance of a point on an external and internal environment, and then was drawn up an experiment in a scaled physical model reduced, where was detected the internal illuminance with three distinct types of facades (a model without any sunscreen, a model with an obstruction blocking black and a model with obstruction blocking plant mass). Analyzing the results obtained, through case studies, it was observed that the vegetation has important features that should be considered during the preparation of a lighting project who have a vegetation as an element of control. This because, like this research verified,a plant mass is able to reflect, transmit and distribute natural light, while filtering the sunlight that affect the environment directly
A comparative analysis of DNA barcode microarray feature size
No full textAbstract Background Microarrays are an invaluable tool in many modern genomic studies. It is generally perceived that decreasing the size of microarray features leads to arrays with higher resolution (due to greater feature density), but this increase in resolution can compromise sensitivity. Results We demonstrate that barcode microarrays with smaller features are equally capable of detecting variation in DNA barcode intensity when compared to larger feature sizes within a specific microarray platform. The barcodes used in this study are the well-characterized set derived from the Yeast KnockOut (YKO) collection used for screens of pooled yeast (Saccharomyces cerevisiae) deletion mutants. We treated these pools with the glycosylation inhibitor tunicamycin as a test compound. Three generations of barcode microarrays at 30, 8 and 5 μm features sizes independently identified the primary target of tunicamycin to be ALG7. Conclusion We show that the data obtained with 5 μm feature size is of comparable quality to the 30 μm size and propose that further shrinking of features could yield barcode microarrays with equal or greater resolving power and, more importantly, higher density.</p
Luz e calor no ambiente construído escolar e o sombreamento arbóreo :: conflito ou compromisso com a conservação de energia ? /
No full textTese (Doutorado) - Universidade Federal de Santa Catarina, Centro Tecnológico
Off-Target Effects of Psychoactive Drugs Revealed by Genome-Wide Assays in Yeast
No full textTo better understand off-target effects of widely prescribed psychoactive drugs, we performed a comprehensive series of chemogenomic screens using the budding yeast Saccharomyces cerevisiae as a model system. Because the known human targets of these drugs do not exist in yeast, we could employ the yeast gene deletion collections and parallel fitness profiling to explore potential off-target effects in a genome-wide manner. Among 214 tested, documented psychoactive drugs, we identified 81 compounds that inhibited wild-type yeast growth and were thus selected for genome-wide fitness profiling. Many of these drugs had a propensity to affect multiple cellular functions. The sensitivity profiles of half of the analyzed drugs were enriched for core cellular processes such as secretion, protein folding, RNA processing, and chromatin structure. Interestingly, fluoxetine (Prozac) interfered with establishment of cell polarity, cyproheptadine (Periactin) targeted essential genes with chromatin-remodeling roles, while paroxetine (Paxil) interfered with essential RNA metabolism genes, suggesting potential secondary drug targets. We also found that the more recently developed atypical antipsychotic clozapine (Clozaril) had no fewer off-target effects in yeast than the typical antipsychotics haloperidol (Haldol) and pimozide (Orap). Our results suggest that model organism pharmacogenetic studies provide a rational foundation for understanding the off-target effects of clinically important psychoactive agents and suggest a rational means both for devising compound derivatives with fewer side effects and for tailoring drug treatment to individual patient genotypes.</p
Multiple means to the same end: the genetic basis of acquired stress resistance in yeast.
No full textIn nature, stressful environments often occur in combination or close succession, and thus the ability to prepare for impending stress likely provides a significant fitness advantage. Organisms exposed to a mild dose of stress can become tolerant to what would otherwise be a lethal dose of subsequent stress; however, the mechanism of this acquired stress tolerance is poorly understood. To explore this, we exposed the yeast gene-deletion libraries, which interrogate all essential and non-essential genes, to successive stress treatments and identified genes necessary for acquiring subsequent stress resistance. Cells were exposed to one of three different mild stress pretreatments (salt, DTT, or heat shock) and then challenged with a severe dose of hydrogen peroxide (H(2)O(2)). Surprisingly, there was little overlap in the genes required for acquisition of H(2)O(2) tolerance after different mild-stress pretreatments, revealing distinct mechanisms of surviving H(2)O(2) in each case. Integrative network analysis of these results with respect to protein-protein interactions, synthetic-genetic interactions, and functional annotations identified many processes not previously linked to H(2)O(2) tolerance. We tested and present several models that explain the lack of overlap in genes required for H(2)O(2) tolerance after each of the three pretreatments. Together, this work shows that acquired tolerance to the same severe stress occurs by different mechanisms depending on prior cellular experiences, underscoring the context-dependent nature of stress tolerance
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