CoREST is an integral component of the CoREST- human histone deacetylase complex

Here we describe the components of a histone deacetylase (HDAC) complex that we term the CoREST-HDAC complex. CoREST-HDAC is composed of polypeptides distinct from previously characterized HDAC1/2-containing complexes such as the mSin3 and nucleosome remodeling and deacetylating (NRD, also named NURD, NuRD) complex. Interestingly, we do not observe RbAp46 and RbAp48 in this complex, although these proteins have been observed in all previously identified complexes and are thought to be part of an HDAC1/2 core. We identify the transcriptional corepressor CoREST and a protein with homology to polyamine oxidases as components of CoREST-HDAC. The HDAC1/2-interacting region of CoREST is mapped to a 179-aa region containing a SANT domain, a domain found in other HDAC1/2-interacting proteins such as NCoR, MTA1, and MTA2. Furthermore, we demonstrate that the corepressor function of CoREST depends on this region. Although CoREST initially was cloned as a corepressor to REST (RE1 silencing transcription factor/neural restrictive silencing factor), we find no evidence for the existence of the eight-zinc finger REST transcription factor as an interacting partner in this complex; however, we do find evidence for association of the putative oncogene ZNF 217 that contains eight zinc fingers.

Protein kinase Cdc15 activates the Dbf2-Mob1 kinase complex

Exit from mitosis in budding yeast requires inactivation of cyclin-dependent kinases through mechanisms triggered by the protein phosphatase Cdc14. Cdc14 activity, in turn, is regulated by a group of proteins, the mitotic exit network (MEN), which includes Lte1, Tem1, Cdc5, Cdc15, Dbf2/Dbf20, and Mob1. The direct biochemical interactions between the components of the MEN remain largely unresolved. Here, we investigate the mechanisms that underlie activation of the protein kinase Dbf2. Dbf2 kinase activity depended on Tem1, Cdc15, and Mob1 in vivo. In vitro, recombinant protein kinase Cdc15 activated recombinant Dbf2, but only when Dbf2 was bound to Mob1. Conserved phosphorylation sites Ser-374 and Thr-544 (present in the human, Caenorhabditis elegans, and Drosophila melanogaster relatives of Dbf2) were required for DBF2 function in vivo, and activation of Dbf2-Mob1 by Cdc15 in vitro. Although Cdc15 phosphorylated Dbf2, Dbf2–Mob1, and Dbf2(S374A/T544A)–Mob1, the pattern of phosphate incorporation into Dbf2 was substantially altered by either the S374A T544A mutations or omission of Mob1. Thus, Cdc15 promotes the exit from mitosis by directly switching on the kinase activity of Dbf2. We propose that Mob1 promotes this activation process by enabling Cdc15 to phosphorylate the critical Ser-374 and Thr-544 phosphoacceptor sites of Dbf2.

A quantitative test to estimate neutralizing antibodies to the hepatitis C virus: cytofluorimetric assessment of envelope glycoprotein 2 binding to target cells.

Hepatitis C virus (HCV) is a major cause of chronic hepatitis. The virus does not replicate efficiently in cell cultures, and it is therefore difficult to assess infection-neutralizing antibodies and to evaluate protective immunity in vitro. To study the binding of the HCV envelope to cell-surface receptors, we developed an assay to assess specific binding of recombinant envelope proteins to human cells and neutralization thereof. HCV recombinant envelope proteins expressed in various systems were incubated with human cells, and binding was assessed by flow cytometry using anti-envelope antibodies. Envelope glycoprotein 2 (E2) expressed in mammalian cells, but not in yeast or insect cells, binds human cells with high affinity (Kd approximately 10(-8) M). We then assessed antibodies able to neutralize E2 binding in the sera of both vaccinated and carrier chimpanzees, as well as in the sera of humans infected with various HCV genotypes. Vaccination with recombinant envelope proteins expressed in mammalian cells elicited high titers of neutralizing antibodies that correlated with protection from HCV challenge. HCV infection does not elicit neutralizing antibodies in most chimpanzees and humans, although low titers of neutralizing antibodies were detectable in a minority of infections. The ability to neutralize binding of E2 derived from the HCV-1 genotype was equally distributed among sera from patients infected with HCV genotypes 1, 2, and 3, demonstrating that binding of E2 is partly independent of E2 hypervariable regions. However, a mouse monoclonal antibody raised against the E2 hypervariable region 1 can partially neutralize binding of E2, indicating that at least two neutralizing epitopes, one of which is hypervariable, should exist on the E2 protein. The neutralization-of-binding assay described will be useful to study protective immunity to HCV infection and for vaccine development.

Spa holístico na esfera urbana portuense: o bem-estar e as leis da natureza

Durante a vida acumula-se, pelos sentidos, um conjunto de sensações necessárias e impreteríveis ao ato de reflexão, por quanto: sem a existência de um reservatório de impressões sensoriais, a mente não tem como refletir sobre elas. Durante a reflexão, temos uma resposta imediata a um estímulo condicionado pelo conhecimento prévio (experiências passadas) e pela identificação deste estímulo com um tempo e espaço em particular. Estas perceções também estão sujeitas aos fatores culturais, portanto, os estereótipos das coisas e eventos do processo de socialização que influenciam fortemente a perceção humana. Ao contrário, a nossa perceção espacial que envolve uma construção gradual e certamente não existe uma perceção previamente feito do nosso desenvolvimento mental. Neste caso, a perceção humana é uma resposta a um campo de informação interrelacionada e não só de fatores de informação individual. A mente organiza esses elementos sensoriais, de modo a formar uma experiência coerente. Os cinco sistemas não trabalham em exclusivo; eles se inter-relacionam recolhendo a mesma informação. Casa sistema percetual colabora com os estímulos parciais entre si e o sistema sensorial com o sistema motor que, em uma constelação fisiológica variável, mantém constante a sensação, o que, portanto, proíbe que o processo nervoso aja como uma simples transmissão de uma mensagem dada. Na época renascentista acreditava-se que os sistemas percetuais estavam relacionados com a imagem do corpo cósmico; a visão estava correlacionada com o fogo e a luz, a audição com o ar, o olfato com o vapor, o gosto com a água e o toque com a terra. Esta intervenção projetual adotou essa relação atribuindo a cada piso uma referência sensorial. Existem várias ciências holísticas que estudam as vertentes que podem influenciar na hora de edificar. Em combinação com os sentidos, os princípios da ciência védica podem delimitar o perímetro construído e as proporções que conferem a harmonia, a base da beleza e a funcionalidade. Deste modo, honram-se as bases da consciência do bem-estar de quem o utilizará. A colaboração do arquiteto ou designer e o utilizador é crucial para determinar as sensações que o utilizador irá encontrar no espaço. com a ajuda de várias referências de obras e autores, propõe-se intervir num edifício localizado na rua de Santa Catarina, entre as ruas Passos Manuel e 31 de Janeiro, na cidade do Porto. O imóvel conta com quatro pisos, com uma cave destinada a arrumos. A proposta de intervenção pretende aplicar a teoria do design sensorial a um projeto de spa de conceito inovador. O utilizador poderá navegar pelo edifício de princípio ao fim, através de diversos tratamentos e terapias, para atingir o bem-estar em cada experiência. O objetivo é despertar os sentidos subjugados pelo frenesim do quotidiano citadino

1994 UM Women's Tennis team

1993 UM Women's Tennis Team

1993 UM Women's Tennis Team

1994 UM Women's Tennis Team

1995 UM Women's Tennis Team

University of Michigan Men's Basketball Team, 2003

Back Row:: academic advisor Angie Heath, head coach Tommy Amaker, director of basketball operations Kirsten Green, asst. coach Billy Schmidt, asst. coach Charles E. Ramsey, Lester Abram, Chuck Bailey, J.C. Mathis, Graham Brown, Chris Hunter, Amadou Ba, Colin Dill, Bernard Robinson Jr., admin. asst. Andy Moore, strength coach Jim Plocki, asst. coach Chuck Swenson, equip. mngr. Bob Bland.

Front Row: trainer Eric Broekhuizen, sr. mngr. Michael Oh, Sherrod Harrell, Dommanic Ingerson, Gavin Groninger, LaVell Blanchard, Rotolu Adebiyi, Daniel Horton, Avery Queen, sr. mngr. Jesse Grasty, sr. mngr. Lucien St. Gerard.

University of Michigan Men's Basketball Team, 2004

Back Row: coach Tommy Amaker, director of basketball operations Kirsten Green, asst. coach Chuck Swenson, asst. coach Charles E. Ramsey, Lester Abram, Brent Petway, Amadou Ba, Chris Hunter, Courtney Sims, Graham Brown, John Andrews, asst. coach Andrew Moore, strength coach Jim Plocki, academic advisor Support Angie Heath, equip. mngr. Bob Bland.

Front Row: trainer Eric Broekhuizen, mngr. Michael Oh, Sherrod Harrell, Daniel Horton, Colin Dill, Bernard Robinson Jr., J.C. Mathis, Dion Harris, Dani Wohl, Manager Matt Duprey, admin. asst. Lucien St. Gerard.

University of Michigan Men's Basketball Team, 2005

Back Row: head coach Tommy Amaker, director of basketball operations Kirsten Green, asst. coach Chuck Swenson, asst. coach Charles Ramsey, John Andrews, Brent Petway, Courtney Sims, Chris Hunter, Amadou Ba, Graham Brown, Ronald Coleman, asst. coach Andrew More academic advisor Angie Beck, strength coach Jim Plocki, equip. mngr. Bob Bland.

Front Row: administrative asst. Lucien St. Gerard, Dani Wohl, Dion Harris, Sherrod Harrell, J.C. Mathis, Lester Abram, Daniel Bell, sr. mngr. Matt Duprey.

1993/1994 UM Women's Basketball Team

Back Row: head coach Trish Roberts, assistant coach Kathy LaBarge Miles, manager Ngina Colston, Catherine DiGiacinto, Jennifer Brzezinski, Rhonda Jokisch, Angie Mustonen, Yeshimbra Gray, manager Danielle Hill, assistant coach Sandy Thomas, assistant coach Carol Owens

Front Row: Tannisha Stevens, Silver Shellman, Amy Johnson, Sonya Mays, Mekisha Ross, Jennifer Keifer

1996/1997 UM Women's Basketball Team

Back Row: ass't coach Eileen Shea, manager Ngina Colston, Molly Murray, Silver Shellman, Shauna Sikorski, Anne Poglits, Catherine DiGiacinto, Tiffany Willard, Kenisha Walker, Pollyanna Johns, ass't coach Yvette Harris

Middle Row: trainer Susan Graham, Amy Johnson, Ann Lemire, Jennifer Kiefer, Akisha Franklin, Mekisha Ross, Stacy Thomas, Jaye Peterson, Angie Phillips

Front Row: ass't coach Yulonda Wimbish, head coach Sue Guevara