Transposon tools for recombinant DNA manipulation: characterization of transcriptional regulators from yeast, Xenopus, and mouse.

Transposon Tn1000 has been adapted to deliver novel DNA sequences for manipulating recombinant DNA. The transposition procedure for these "tagged" Tn1000s is simple and applicable to most plasmids in current use. For yeast molecular biology, tagged Tn1000s introduce a variety of yeast selective markers and replication origins into plasmids and cosmids. In addition, the beta-globin minimal promoter and lacZ gene of Tn(beta)lac serve as a mobile reporter of eukaryotic enhancer activity. In this paper, Tn(beta)lac was used to localize a mouse HoxB-complex enhancer in transgenic mice. Other tagged transposons create Gal4 DNA-binding-domain fusions, in either Escherichia coli or yeast plasmids, for use in one- and two-hybrid tests of transcriptional activation and protein-protein interaction, respectively. With such fusions, the Saccharomyces cerevisiae Swi6 G1/S-phase transcription factor and the Xenopus laevis Pintallavis developmental regulator are shown to activate transcription. Furthermore, the same transposon insertions also facilitated mapping of the Swi6 and Pintallavis domains responsible for transcriptional activation. Thus, as well as introducing novel sequences, tagged transposons share the numerous other applications of transposition such as producing insertional mutations, creating deletion series, or serving as mobile primer sites for DNA sequencing.

Postseptational chromosome partitioning in bacteria.

Mutations in the spoIIIE gene prevent proper partitioning of one chromosome into the developing prespore during sporulation but have no overt effect on partitioning in vegetatively dividing cells. However, the expression of spoIIIE in vegetative cells and the occurrence of genes closely related to spoIIIE in a range of nonsporulating eubacteria suggested a more general function for the protein. Here we show that SpoIIIE protein is needed for optimal chromosome partitioning in vegetative cells of Bacillus subtilis when the normal tight coordination between septation and nucleoid partitioning is perturbed or when septum positioning is altered. A functional SpoIIIE protein allows cells to recover from a state in which their chromosome has been trapped by a closing septum. By analogy to its function during sporulation, we suggest that SpoIIIE facilitates partitioning by actively translocating the chromosome out of the septum. In addition to enhancing the fidelity of nucleoid partitioning, SpoIIIE also seems to be required for maximal resistance to antibiotics that interfere with DNA metabolism. The results have important implications for our understanding of the functions of genes involved in the primary partitioning machinery in bacteria and of how septum placement is controlled.

Developing Civil Society and a New Leadership Class in Iraq: A Comprehensive Analysis and Business Plan

Following De-Ba'athification, forming a new leadership class will be critical to the success of creating a strong civil society in modern-day Iraq. Implementation of youth educational exchange programs, specifically promoting leadership skills, is a significant part of the solution to stimulating a new generation of leaders. Using the reign of Saddam Hussein and his toppling as a frame of reference, a brief history of Iraq's civil society reveals a need for a new leadership class through the lens of democratic transition and consolidation. After exploring the leadership challenges of post-war nation building, the proposed business plan focuses on implementing a youth leadership program in Iraq, employing a wider participation model, and a lengthier, more involved learning model from existing programs.

Contarlo como es

Las historias transmitidas oralmente constituyen un medio para incluir de forma productiva a las personas desplazadas forzadamente, mediante sus opiniones, en el trabajo y las prácticas de quienes buscan soluciones para las crisis de desplazamiento.

Biometrika.

v.2 (1902-1903)

Efeito das taxas moderadoras no consumo de cuidados de saúde primários

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Fatores de risco para a progressão da gamapatia monoclonal de significado indeterminado para mieloma múltiplo

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Taquicardia ventricular idiopática : estudo de correlação do ECG com o local de origem

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Comparação de modelos estáticos, otimização dos determinantes da estrutura de capital das PME's portuguesas

Dissertação, Mestrado, Contabilidade e Fiscalidade, Instituto Politécnico de Santarém, Escola Superior de Gestão e Tecnologia, 2016

Imunosenescência

Dissertação para obtenção do grau de Mestre no Instituto Superior de Ciências da Saúde Egas Moniz

Light sheet fluorescence microscopy for in situ cell interaction analysis in mouse lymph nodes.

Reactive lymph nodes (LNs) are sites where pMHC-loaded dendritic cells (DCs) interact with rare cognate T cells, leading to their clonal expansion. While DC interactions with T cell subsets critically shape the ensuing immune response, surprisingly little is known on their spatial orchestration at physiologically T cell low precursor frequencies. Light sheet fluorescence microscopy and one of its implementations, selective plane illumination microscopy (SPIM), is a powerful method to obtain precise spatial information of entire organs of 0.5-10mm diameter, the size range of murine LNs. Yet, its usefulness for immunological research has thus far not been comprehensively explored. Here, we have tested and defined protocols that preserve fluorescent protein function during lymphoid tissue clearing required for SPIM. Reconstructions of SPIM-generated 3D data sets revealed that calibrated numbers of adoptively transferred T cells and DCs are successfully detected at a single cell level within optically cleared murine LNs. Finally, we define parameters to quantify specific interactions between antigen-specific T cells and pMHC-bearing DCs in murine LNs. In sum, our studies describe the successful application of light sheet fluorescence microscopy to immunologically relevant tissues.

Commemoração do centenário da Praça do Commercio do Pará, 1819-1919 /

Cover title.

1966 UM football players from Chicago area

back row L-R: Royce Spencer, Robert Baumgartner, Robert Kieta, Robert Mielke, David Fisher, Ray Phillips. front row L-R: John Lynch, Gerald Miklos, Ernest Sharpe, Richard Yanz,

UM Women's Track Team, 1979

Back Row: Dede Key, Julie Clifford, Tina Smith, Debbie Williams, Penny Neer, Sherrie King, coach Red Simmons

Front Row: Pam Moore, Kathy Kampen, Dawn Woodruff, Catherine Sharpe,

UM Women's Track Team, 1980

Front Row (left to right): Cathy Sharpe, Brenda Kazinec, Renee Turner, Lorrie Thornton, Dawn Woodruff, Sue Frederick, Julie Clifford.

Middle Row: Coach Ken Simmons, Kathy Kampen, Tina Smith, Cornelia Kaufmann, Nanette Feleccia, Maureen Miner, Dana Loesche, Kathy Benner, Sheila Mayberry.

Back Row: Assistant Coach Scott Hubbard, Deb Williams, Janet Hallfriach, Karen Perry, Joanna Bullard, Melanie Weaver, Sheila Shatter

Missing: Lynn Fudala, Dede Key, Sherrie King, Penny Neer, Sharon Wigglesworth.