Intercellular adhesion and membrane polarity in rat hepatocytes: Localization of cell -CAM 105 and other domain-specific membrane proteins {\it in situ\/} and {\it in vitro\/} by electron microscopy

Cell-CAM 105 has been identified as a cell adhesion molecule (CAM) based on the ability of monospecific and monovalent anti-cell-CAM 105 antibodies to inhibit the reaggregation of rat hepatocytes. Although one would expect to find CAMs concentrated in the lateral membrane domain where adhesive interactions predominate, immunofluorescence analysis of rat liver frozen sections revealed that cell-CAM 105 was present exclusively in the bile canalicular (BC) domain of the hepatocyte. To more precisely define the in situ localization of cell-CAM 105, immunoperoxidase and electron microscopy were used to analyze intact and mechanically dissociated fixed liver tissue. Results indicate that although cell-CAM 105 is apparently restricted to the BC domain in situ, it can be detected in the pericanalicular region of the lateral membranes when accessibility to lateral membranes is provided by mechanical dissociation. In contrast, when hepatocytes were labeled following incubation in vitro under conditions used during adhesion assays, cell-CAM 105 had redistributed to all areas of the plasma membrane. Immunofluorescence analysis of primary hepatocyte cultures revealed that cell-CAM 105 and two other BC proteins were localized in discrete domains reminscent of BC while cell-CAM 105 was also present in regions of intercellular contact. These results indicate that the distribution of cell-CAM 105 under the experimental conditions used for cell adhesion assays differs from that in situ and raises the possibility that its adhesive function may be modulated by its cell surface distribution. The implications of these and other findings are discussed with regard to a model for BC formation.^ Analysis of molecular events involved in BC formation would be accelerated if an in vitro model system were available. Although BC formation in culture has previously been observed, repolarization of cell-CAM 105 and two other domain-specific membrane proteins was incomplete. Since DMSO had been used by Isom et al. to maintain liver-specific gene expression in vitro, the effect of this differentiation system on the polarity of these membrane proteins was examined. Based on findings presented here, DMSO apparently prolongs the expression and facilitates polarization of hepatocyte membrane proteins in vitro. ^

Climas motivacionales, liderazgo y cohesión grupal en contexto deportivo universitario

Objetivos: Conocer las interacciones entrenador-atleta y comprender las practicas de liderazgo, de generación de climas motivacionales y de comunicación entre líderes deportivos y jugadores. Metodología: Metodología de cohorte mixto. Se realizó correlación de variables cuantitativas y se analizaron las experiencias y los sentidos del contexto deportivo con una aproximación cualitativa. Se aplicaron a 31 deportistas universitarios los instrumentos: Clima Motivacional Percibido en el Deporte (PMCSQ-2), Clima en el Deporte (SCQ) y Orientación al Ego y a la Tarea en el Deporte. Para profundizar la información obtenida, se realizaron entrevistas semi-estructuradas a 6 deportistas y 2 entrenadores universitarios. RESULTADOS: Que el deportista sienta confianza en su entrenador se encuentra asociado a que se sienta comprendido y aceptado por él. Que el entrenador genere un clima motivacional orientado hacia el ego está relacionado con que los deportistas tengan orientaciones de meta ego. Los entrenadores utilizan dos estilos de liderazgo opuestos: liderazgo democrático (entrenamientos) y liderazgo autocrático (competiciones) Conclusiones: Cuando el atleta confía en la persona que lo dirige deportivamente, presenta mayor satisfacción deportiva. También, que el entrenador fomente un ambiente de comparación social propicia que los deportistas rivalicen con compañeros de equipo y basen su rendimiento en resultados deportivos obtenidos

Climas motivacionales, liderazgo y cohesión grupal en contexto deportivo universitario

Objetivos: Conocer las interacciones entrenador-atleta y comprender las practicas de liderazgo, de generación de climas motivacionales y de comunicación entre líderes deportivos y jugadores. Metodología: Metodología de cohorte mixto. Se realizó correlación de variables cuantitativas y se analizaron las experiencias y los sentidos del contexto deportivo con una aproximación cualitativa. Se aplicaron a 31 deportistas universitarios los instrumentos: Clima Motivacional Percibido en el Deporte (PMCSQ-2), Clima en el Deporte (SCQ) y Orientación al Ego y a la Tarea en el Deporte. Para profundizar la información obtenida, se realizaron entrevistas semi-estructuradas a 6 deportistas y 2 entrenadores universitarios. RESULTADOS: Que el deportista sienta confianza en su entrenador se encuentra asociado a que se sienta comprendido y aceptado por él. Que el entrenador genere un clima motivacional orientado hacia el ego está relacionado con que los deportistas tengan orientaciones de meta ego. Los entrenadores utilizan dos estilos de liderazgo opuestos: liderazgo democrático (entrenamientos) y liderazgo autocrático (competiciones) Conclusiones: Cuando el atleta confía en la persona que lo dirige deportivamente, presenta mayor satisfacción deportiva. También, que el entrenador fomente un ambiente de comparación social propicia que los deportistas rivalicen con compañeros de equipo y basen su rendimiento en resultados deportivos obtenidos

Climas motivacionales, liderazgo y cohesión grupal en contexto deportivo universitario

Objetivos: Conocer las interacciones entrenador-atleta y comprender las practicas de liderazgo, de generación de climas motivacionales y de comunicación entre líderes deportivos y jugadores. Metodología: Metodología de cohorte mixto. Se realizó correlación de variables cuantitativas y se analizaron las experiencias y los sentidos del contexto deportivo con una aproximación cualitativa. Se aplicaron a 31 deportistas universitarios los instrumentos: Clima Motivacional Percibido en el Deporte (PMCSQ-2), Clima en el Deporte (SCQ) y Orientación al Ego y a la Tarea en el Deporte. Para profundizar la información obtenida, se realizaron entrevistas semi-estructuradas a 6 deportistas y 2 entrenadores universitarios. RESULTADOS: Que el deportista sienta confianza en su entrenador se encuentra asociado a que se sienta comprendido y aceptado por él. Que el entrenador genere un clima motivacional orientado hacia el ego está relacionado con que los deportistas tengan orientaciones de meta ego. Los entrenadores utilizan dos estilos de liderazgo opuestos: liderazgo democrático (entrenamientos) y liderazgo autocrático (competiciones) Conclusiones: Cuando el atleta confía en la persona que lo dirige deportivamente, presenta mayor satisfacción deportiva. También, que el entrenador fomente un ambiente de comparación social propicia que los deportistas rivalicen con compañeros de equipo y basen su rendimiento en resultados deportivos obtenidos

Climas motivacionales, liderazgo y cohesión grupal en contexto deportivo universitario

Objetivos: Conocer las interacciones entrenador-atleta y comprender las practicas de liderazgo, de generación de climas motivacionales y de comunicación entre líderes deportivos y jugadores. Metodología: Metodología de cohorte mixto. Se realizó correlación de variables cuantitativas y se analizaron las experiencias y los sentidos del contexto deportivo con una aproximación cualitativa. Se aplicaron a 31 deportistas universitarios los instrumentos: Clima Motivacional Percibido en el Deporte (PMCSQ-2), Clima en el Deporte (SCQ) y Orientación al Ego y a la Tarea en el Deporte. Para profundizar la información obtenida, se realizaron entrevistas semi-estructuradas a 6 deportistas y 2 entrenadores universitarios. RESULTADOS: Que el deportista sienta confianza en su entrenador se encuentra asociado a que se sienta comprendido y aceptado por él. Que el entrenador genere un clima motivacional orientado hacia el ego está relacionado con que los deportistas tengan orientaciones de meta ego. Los entrenadores utilizan dos estilos de liderazgo opuestos: liderazgo democrático (entrenamientos) y liderazgo autocrático (competiciones) Conclusiones: Cuando el atleta confía en la persona que lo dirige deportivamente, presenta mayor satisfacción deportiva. También, que el entrenador fomente un ambiente de comparación social propicia que los deportistas rivalicen con compañeros de equipo y basen su rendimiento en resultados deportivos obtenidos

HLA alleles determine human T-lymphotropic virus-I (HTLV-I) proviral load and the risk of HTLV-I-associated myelopathy

The risk of disease associated with persistent virus infections such as HIV-I, hepatitis B and C, and human T-lymphotropic virus-I (HTLV-I) is strongly determined by the virus load. However, it is not known whether a persistent class I HLA-restricted antiviral cytotoxic T lymphocyte (CTL) response reduces viral load and is therefore beneficial or causes tissue damage and contributes to disease pathogenesis. HTLV-I-associated myelopathy (HAM/TSP) patients have a high virus load compared with asymptomatic HTLV-I carriers. We hypothesized that HLA alleles control HTLV-I provirus load and thus influence susceptibility to HAM/TSP. Here we show that, after infection with HTLV-I, the class I allele HLA-A*02 halves the odds of HAM/TSP (P < 0.0001), preventing 28% of potential cases of HAM/TSP. Furthermore, HLA-A*02+ healthy HTLV-I carriers have a proviral load one-third that (P = 0.014) of HLA-A*02− HTLV-I carriers. An association of HLA-DRB1*0101 with disease susceptibility also was identified, which doubled the odds of HAM/TSP in the absence of the protective effect of HLA-A*02. These data have implications for other persistent virus infections in which virus load is associated with prognosis and imply that an efficient antiviral CTL response can reduce virus load and so prevent disease in persistent virus infections.

Induction of Purkinje fiber differentiation by coronary arterialization

A synchronized heart beat is controlled by pacemaking impulses conducted through Purkinje fibers. In chicks, these impulse-conducting cells are recruited during embryogenesis from myocytes in direct association with developing coronary arteries. In culture, the vascular cytokine endothelin converts embryonic myocytes to Purkinje cells, implying that selection of conduction phenotype may be mediated by an instructive cue from arteries. To investigate this hypothesis, coronary arterial development in the chicken embryo was either inhibited by neural crest ablation or activated by ectopic expression of fibroblast growth factor (FGF). Ablation of cardiac neural crest resulted in ≈70% reductions (P < 0.01) in the density of intramural coronary arteries and associated Purkinje fibers. Activation of coronary arterial branching was induced by retrovirus-mediated overexpression of FGF. At sites of FGF-induced hypervascularization, ectopic Purkinje fibers differentiated adjacent to newly induced coronary arteries. Our data indicate the necessity and sufficiency of developing arterial bed for converting a juxtaposed myocyte into a Purkinje fiber cell and provide evidence for an inductive function for arteriogenesis in heart development distinct from its role in establishing coronary blood circulation.

The neuropeptide Y/agouti gene-related protein (AGRP) brain circuitry in normal, anorectic, and monosodium glutamate-treated mice

Neuropeptide Y (NPY) and the endogenous melanocortin receptor antagonist, agouti gene-related protein (AGRP), coexist in the arcuate nucleus, and both exert orexigenic effects. The present study aimed primarily at determining the brain distribution of AGRP. AGRP mRNA-expressing cells were limited to the arcuate nucleus, representing a major subpopulation (95%) of the NPY neurons, which also was confirmed with immunohistochemistry. AGRP-immunoreactive (-ir) terminals all contained NPY and were observed in many brain regions extending from the rostral telencephalon to the pons, including the parabrachial nucleus. NPY-positive, AGRP-negative terminals were observed in many areas. AGRP-ir terminals were reduced dramatically in all brain regions of mice treated neonatally with monosodium glutamate as well as of mice homozygous for the anorexia mutation. Terminals immunoreactive for the melanocortin peptide α-melanocyte-stimulating hormone formed a population separate from, but parallel to, the AGRP-ir terminals. Our results show that arcuate NPY neurons, identified by the presence of AGRP, project more extensively in the brain than previously known and indicate that the feeding regulatory actions of NPY may extend beyond the hypothalamus.

Differential regulation of transcription: Repression by unactivated mitogen-activated protein kinase Kss1 requires the Dig1 and Dig2 proteins

Kss1, a yeast mitogen-activated protein kinase (MAPK), in its unphosphorylated (unactivated) state binds directly to and represses Ste12, a transcription factor necessary for expression of genes whose promoters contain filamentous response elements (FREs) and genes whose promoters contain pheromone response elements (PREs). Herein we show that two nuclear proteins, Dig1 and Dig2, are required cofactors in Kss1-imposed repression. Dig1 and Dig2 cooperate with Kss1 to repress Ste12 action at FREs and regulate invasive growth in a naturally invasive strain. Kss1-imposed Dig-dependent repression of Ste12 also occurs at PREs. However, maintenance of repression at PREs is more dependent on Dig1 and/or Dig2 and less dependent on Kss1 than repression at FREs. In addition, derepression at PREs is more dependent on MAPK-mediated phosphorylation than is derepression at FREs. Differential utilization of two types of MAPK-mediated regulation (binding-imposed repression and phosphorylation-dependent activation), in combination with distinct Ste12-containing complexes, contributes to the mechanisms by which separate extracellular stimuli that use the same MAPK cascade can elicit two different transcriptional responses.

Immunization of metastatic breast cancer patients with a fully synthetic globo H conjugate: A phase I trial

The carbohydrate antigen globo H commonly found on breast cancer cells is a potential target for vaccine therapy. The objectives of this trial were to determine the toxicity and immunogenicity of three synthetic globo H-keyhole limpet hemocyanin conjugates plus the immunologic adjuvant QS-21. Twenty-seven metastatic breast cancer patients received five vaccinations each. The vaccine was well tolerated, and no definite differences were observed among the three formulations. Serologic analyses demonstrated the generation of IgM antibody titers in most patients, with minimal IgG antibody stimulation. There was significant binding of IgM antibodies to MCF-7 tumor cells in 16 patients, whereas IgG antibody reactivity was observed in a few patients. There was evidence of complement-dependent cytotoxicity in several patients. Affinity column purification supported the specificity of IgM antibodies for globo H. On the basis of these data, globo H will constitute one component of a polyvalent vaccine for evaluation in high-risk breast cancer patients.

A role for estrogen receptor β in the regulation of growth of the ventral prostate

In normal rats and mice, immunostaining with specific antibodies revealed that nuclei of most prostatic epithelial cells harbor estrogen receptor β (ERβ). In rat ventral prostate, 530- and 549-aa isoforms of the receptor were identified. These sediment in the 4S region of low-salt sucrose gradients, indicating that prostatic ERβ does not contain the same protein chaperones that are associated with ERα. Estradiol (E2) binding and ERβ immunoreactivity coincide on the gradient, with no indication of ERα. In prostates from mice in which the ERβ gene has been inactivated (BERKO), androgen receptor (AR) levels are elevated, and the tissue contains multiple hyperplastic foci. Most epithelial cells express the proliferation antigen Ki-67. In contrast, prostatic epithelium from wild-type littermates is single layered with no hyperplasia, and very few cells express Ki-67. Rat ventral prostate contains an estrogenic component, which comigrates on HPLC with the testosterone metabolite 5α-androstane-3β,17β-diol (3βAdiol). This compound, which competes with E2 for binding to ERβ and elicits an estrogenic response in the aorta but not in the pituitary, decreases the AR content in prostates of wild-type mice but does not affect the elevated levels seen in ERβ knockout (BERKO) mice. Thus ERβ, probably as a complex with 3βAdiol, is involved in regulating the AR content of the rodent prostate and in restraining epithelial growth. These findings suggest that ligands specific for ERβ may be useful in the prevention and/or clinical management of prostatic hyperplasia and neoplasia.

Induction of Acclimative Proteolysis of the Light-Harvesting Chlorophyll a/b Protein of Photosystem II in Response to Elevated Light Intensities1

Most plants have the ability to respond to fluctuations in light to minimize damage to the photosynthetic apparatus. A proteolytic activity has been discovered that is involved in the degradation of the major light-harvesting chlorophyll a/b-binding protein of photosystem II (LHCII) when the antenna size of photosystem II is reduced upon acclimation of plants from low to high light intensities. This ATP-dependent proteolytic activity is of the serine or cysteine type and is associated with the outer membrane surface of the stroma-exposed thylakoid regions. The identity of the protease is not known, but it does not correspond to the recently identified chloroplast ATP-dependent proteases Clp and FtsH, which are homologs to bacterial enzymes. The acclimative response shows a delay of 2 d after transfer of the leaves to high light. This lag period was shown to be attributed to expression or activation of the responsible protease. Furthermore, the LHCII degradation was found to be regulated at the substrate level. The degradation process involves lateral migration of LHCII from the appressed to the nonappressed thylakoid regions, which is the location for the responsible protease. Phosphorylated LHCII was found to be a poor substrate for degradation in comparison with the unphosphorylated form of the protein. The relationship between LHCII degradation and other regulatory proteolytic processes in the thylakoid membrane, such as D1-protein degradation, is discussed.

Commercial work and trainng for girls

by Jeannette Eaton... [and] Bertha M. Stevens...

Forecasting trade and the merchant fleet. Final report.

Maritime Administration, Washington, D.C.