The Yck2 Yeast Casein Kinase 1 Isoform Shows Cell Cycle-specific Localization to Sites of Polarized Growth and Is Required for Proper Septin Organization

Casein kinase 1 protein kinases are ubiquitous and abundant Ser/Thr-specific protein kinases with activity on acidic substrates. In yeast, the products of the redundant YCK1 and YCK2 genes are together essential for cell viability. Mutants deficient for these proteins display defects in cellular morphogenesis, cytokinesis, and endocytosis. Yck1p and Yck2p are peripheral plasma membrane proteins, and we report here that the localization of Yck2p within the membrane is dynamic through the cell cycle. Using a functional green fluorescent protein (GFP) fusion, we have observed that Yck2p is concentrated at sites of polarized growth during bud morphogenesis. At cytokinesis, GFP–Yck2p becomes associated with a ring at the bud neck and then appears as a patch of fluorescence, apparently coincident with the dividing membranes. The bud neck association of Yck2p at cytokinesis does not require an intact septin ring, and septin assembly is altered in a Yck-deficient mutant. The sites of GFP–Yck2p concentration and the defects observed for Yck-deficient cells together suggest that Yck plays distinct roles in morphogenesis and cytokinesis that are effected by differential localization.

Characterization of a replication-incompetent adenovirus type 5 mutant deleted for the preterminal protein gene

An adenovirus type 5 mutant deleted for the preterminal protein (pTP) gene was constructed using cell lines that express pTP. The pTP deletion mutant virus is incapable of replicating in the absence of complementation and does not express detectable levels of viral mRNAs that are expressed only after the onset of replication. Accumulation of early-region mRNAs, including that for E1A, exhibits a lag relative to that observed from the wild-type virus. However, E1A mRNA accumulation attains a steady-state level similar to the level of expression during the early phase of infection with the wild-type virus. In 293-pTP cells (human embryonic kidney cells that express pTP in addition to high levels of adenovirus E1A and E1B proteins), the pTP deletion mutant virus replicates efficiently and yields infectious titers within 5-fold of that of the wild-type virus. The deletion of 1.2 kb of pTP-encoding sequence increases the size of foreign DNA that can be introduced into the virus and, with an absolute block to replication, makes this virus an important tool for gene therapy.

Information needs of clinical teams: analysis of questions received by the Clinical Informatics Consult Service

Objectives: To examine the types of questions received by Clinical Informatics Consult Service (CICS) librarians from clinicians on rounds and to analyze the number of clearly differentiated viewpoints provided in response.

CML66, a broadly immunogenic tumor antigen, elicits a humoral immune response associated with remission of chronic myelogenous leukemia

This report describes a tumor-associated antigen, termed CML66, initially cloned from a chronic myelogenous leukemia (CML) cDNA expression library. CML66 encodes a 583-aa protein with a molecular mass of 66 kDa and no significant homology to other known genes. CML66 gene is localized to human chromosome 8q23, but the function of this gene is unknown. CML66 is expressed in leukemias and a variety of solid tumor cell lines. When examined by Northern blot, expression in normal tissues was restricted to testis and heart, and no expression was found in hematopoietic tissues. When examined by quantitative reverse transcription–PCR, expression in CML cells was 1.5-fold higher than in normal peripheral blood mononuclear cells. The presence of CML66-specific antibody in patient serum was confirmed by Western blot and the development of high titer IgG antibody specific for CML66 correlated with immune induced remission of CML in a patient who received infusion of normal donor lymphocytes for treatment of relapse. CML66 antibody also was found in sera from 18–38% of patients with lung cancer, melanoma, and prostate cancer. These findings suggest that CML66 may be immunogenic in a wide variety of malignancies and may be a target for antigen-specific immunotherapy.

Cloning and characterization of ERG25, the Saccharomyces cerevisiae gene encoding C-4 sterol methyl oxidase.

We have cloned the Saccharomyces cerevisiae C-4 sterol methyl oxidase ERG25 gene. The sterol methyl oxidase performs the first of three enzymic steps required to remove the two C-4 methyl groups leading to cholesterol (animal), ergosterol (fungal), and stigmasterol (plant) biosynthesis. An ergosterol auxotroph, erg25, which fails to demethylate and concomitantly accumulates 4,4-dimethylzy-mosterol, was isolated after mutagenesis. A complementing clone consisting of a 1.35-kb Dra I fragment encoded a 309-amino acid polypeptide (calculated molecular mass, 36.48 kDa). The amino acid sequence shows a C-terminal endoplasmic reticulum retrieval signal KKXX and three histidine-rich clusters found in eukaryotic membrane desaturases and in a bacterial alkane hydroxylase and xylene monooxygenase. The sterol profile of an ERG25 disruptant was consistent with the erg25 allele obtained by mutagenesis.

Transforming growth factor beta mediates the progesterone suppression of an epithelial metalloproteinase by adjacent stroma in the human endometrium.

Unlike most normal adult tissues, cyclic growth and tissue remodeling occur within the uterine endometrium throughout the reproductive years. The matrix metalloproteinases (MMPs), a family of structurally related enzymes that degrade specific components of the extracellular matrix are thought to be the physiologically relevant mediators of extracellular matrix composition and turnover. Our laboratory has identified MMPs of the stromelysin family in the cycling human endometrium, implicating these enzymes in mediating the extensive remodeling that occurs in this tissue. While the stromelysins are expressed in vivo during proliferation-associated remodeling and menstruation-associated endometrial breakdown, none of the stromelysins are expressed during the progesterone-dominated secretory phase of the cycle. Our in vitro studies of isolated cell types have confirmed progesterone suppression of stromal MMPs, but a stromal-derived paracrine factor was found necessary for suppression of the epithelial-specific MMP matrilysin. In this report, we demonstrate that transforming growth factor beta (TGF-beta) is produced by endometrial stroma in response to progesterone and can suppress expression of epithelial matrilysin independent of progesterone. Additionally, we find that an antibody directed against the mammalian isoforms of TGF-beta abolishes progesterone suppression of matrilysin in stromal-epithelial cocultures, implicating TGF-beta as the principal mediator of matrilysin suppression in the human endometrium.

Antropología educativa aplicada a las prácticas clínicas en Enfermería: las diferencias culturales implicadas en el proceso de prácticas clínicas en Enfermería. Una aportación desde la antropología de la complejidad y el pensamiento crítico

En los estudios de la Diplomatura de Enfermería existe una división, tanto a nivel secuencial-cronológico (transmisión gradual y lógica de los contenidos), como en la dimensión espacio-ambiental (aula, sala simulación y servicio sanitario: comunitario u hospitalario). Estas divisiones tienen consecuencias en la estructura curricular de la disciplina: módulo teórico, módulo práctico y módulo práctico-clínico, confiriéndole a estos estudios una peculiaridad asentada en las diferencias en los tipos de escenarios, ambientes, instituciones, normas, y personas que se manifiestan en los contrastes percibidos (Merleau Ponty, 1988) por los alumnos en su proceso de enseñanza-aprendizaje según se dé éste en un módulo u otro. La enorme variabilidad de la práctica se ve también afectada por las características del escenario en el que tienen lugar las prácticas clínicas: centros de atención primaria y/o instituciones hospitalarias, en los que el alumno debe iniciar sus prácticas de enfermería con el que va a constituir su objeto-sujeto de trabajo durante toda su vida profesional: el ser humano (individuo, familia y comunidad) atendiendo a sus necesidades fisiológicas, psicológicas y sociales desde una perspectiva holística. Los contrastes entre el aula y el servicio sanitario suponen a veces contradicciones que desenfocan la visión lógica con la que el alumno pretende interpretar su realidad formativa en esta compleja fase de sus estudios, y esta visión borrosa puede derivar en situaciones de disminución o pérdida de significado que esta experiencia educativa debe tener en el alumno (Bruner, 1991).

Mathematical notebook, and commonplace book, circa 1689-1769

Manuscript notebook, possibly kept by Harvard students, containing 17th century English transcriptions of arithmetic and geometry texts, one of which is dated 1689-1690; 18th century transcriptions from John Ward’s “The Young Mathematician’s Guide”; and notes on physics lectures delivered by John Winthrop, the Hollis Professor of Mathematics and Natural Philosophy at Harvard from 1738 to 1779. The notebook also contains 18th century reading notes on Henry VIII, Tudor succession, and English history from Daniel Neal’s “The History of the Puritans” and David Hume’s “History of England,” and notes on Ancient history, taken mainly from Charles Rollin’s “The Ancient History of the Egyptians, Carthaginians, Assyrians, Babylonians, Medes and Persians, Macedonians and Grecians.” Additionally included are an excerpt from Plutarch’s “Lives” and transcriptions of three articles from “The Gentleman’s Magazine, and Historical Chronicle,” published in 1769: “A Critique on the Works of Ovid”; a book review of “A New Voyage to the West-Indies”; and “Genuine Anecdotes of Celebrated Writers, &.” The flyleaf contains the inscription “Semper boni aliquid operis facito ut diabolus te semper inveniat occupatum,” a variation on a quote of Saint Jerome that translates approximately as “Always good to do some work so that the devil may always find you occupied.” In the seventeenth and eighteenth centuries, Harvard College undergraduates often copied academic texts and lecture notes into personal notebooks in place of printed textbooks. Winthrop used Ward’s textbook in his class, while the books of Hume, Neal, and Rollin were used in history courses taught at Harvard in the 18th century.

Tracing the Origins of Athlete Development Models in Sport: a Citation Path Analysis

Reviews of the sport psychology literature have identified a number of models of athlete development in sport (Alfermann & Stambulova, 2007; Durand-Bush &Salmela, 2001). However, minimal research has investigated the origins of knowledge from which each model was developed. The purpose of this study was to systematically examine the influential texts responsible for providing the basis of athlete development models in sport. A citation path analysis of the sport psychology literature was used to generate a knowledge development path of seven athlete development models in sport. The analysis identified influential texts and authors in the conceptualization of athlete development. The popula-tion of 229 texts (articles, books, book chapters) was selected in two phases. Phase1 texts were articles citing seven articles depicting models of athlete development(n  75). Phase 2 included texts cited three or more times by Phase 1 articles (n  154). The analysis revealed how the scholarship of Benjamin Bloom (1985) has been integrated into the field of sport psychology, and how two articles appearing in 1993 and 2003 helped shape present conceptualizations of athlete development

Gaining Insight into Actual and Preferred Sources of Coaching Knowledge

Previous research has suggested that current formal coach education programs do not fully meet the learning needs of coaches. The purpose of the present study was to examine actual and preferred sources of coaching knowledge for developmental-level coaches. Structured quantitative interviews were conducted with coaches (N = 44) from a variety of sports. Learning by doing, interaction with coaching peers, and formal coach education were the top actual sources of coaching knowledge. Discrepancies were found between actual and preferred usage of learning by doing, formal coach education, and mentoring. Coaches indicated they would prefer more guided learning and less self-directed learning by doing. Further, differences in preferred sources were identified between coaches wishing to move to an elite level versus coaches wishing to stay at a developmental level. Findings highlight the importance of both experiential and formally guided sources of coaching knowledge and the context-specific nature of coach learning.