Gp74 a membrane glycoprotein of the cis-Golgi network that cycles through the endoplasmic reticulum and intermediate compartment

A monoclonal antibody CC92 (IgM), raised against a fraction of rat liver enriched in Golgi membranes, recognizes a novel Endo H-resistant 74-kD membrane glycoprotein (gp74). The bulk of gp74 is confined to the cis-Golgi network (CGN). Outside the Golgi gp74 is found in tubulovesicular structures and ER foci. In cells incubated at 37 degrees C the majority of gp74 is segregated from the intermediate compartment (IC) marker p58. However, in cells treated with organelle perturbants such as low temperature, BFA, and [AIF4]- the patterns of the two proteins become indistinguishable. Both proteins are retained in the Golgi complex at 20 degrees C and in the IC at 15 degrees C. Incubation of cells with BFA results in relocation of gp74 to p58 positive IC elements. [AIF4]- induces the redistribution of gp74 from the Golgi to p58-positive vesicles and does not retard the translocation of gp74 to IC elements in cells treated with BFA. Disruption of microtubules by nocodazol results in the rapid disappearance of the Golgi elements stained by gp74 and redistribution of the protein into vesicle-like structures. The responses of gp74 to cell perturbants are in sharp contrast with those of cis/middle and trans-Golgi resident proteins whose location is not affected by low temperatures or [AIF4]-, are translocated to the ER upon addition of BFA, and stay in slow disintegrating Golgi elements in cells treated with nocodazol. The results suggest that gp74 is an itinerant protein that resides most of the time in the CGN and cycles through the ER/IC following the pathway used by p58.

Developmental expression of the oligodendrocyte myelin glycoprotein in the mouse telencephalon

The oligodendrocyte myelin glycoprotein is a glycosylphosphatidylinositol-anchored protein expressed by neurons and oligodendrocytes in the CNS. Attempts have been made to identify the functions of the myelin-associated inhibitory proteins (MAIPs) after axonal lesion or in neurodegeneration. However, the developmental roles of some of these proteins and their receptors remain elusive. Recent studies indicate that NgR1 and the recently discovered receptor PirB restrict cortical synaptic plasticity. However, the putative factors that trigger these effects are unknown. Since Nogo-A is mostly associated with the endoplasmic reticulum and MAG appears late during development, the putative participation of OMgp should be considered. Here we examine the pattern of development of OMgp immunoreactive elements during mouse telencephalic development. OMgp immunoreactivity in the developing cortex follows the establishment of the thalamo-cortical barrel-field. At cellular level, we located OMgp neuronal membranes in dendrites and axons as well as in brain synaptosome fractions and axon varicosities. Lastly, the analysis of the barrel-field in OMgp-deficient mice revealed that although thalamo-cortical connections were formed, their targeting in layer IV was altered and numerous axons ectopically invaded layer II-III. Our data support the idea that early-expressed MAIPs play an active role during development and point to OMgp participating in thalamo-cortical connections.

Different Storing and Processing Conditions of Human Lymphocytes do not Alter P-Glycoprotein Rhodamine 123 Efflux.

P-glycoprotein (Pgp), a protein codified by Multi Drug Resistance (MDR1) gene, has a detoxifying function and might influence the toxicity and pharmacokinetics and pharmacodynamics of drugs. Sampling strategies to improve Pgp studies could be useful to optimize the sensitivity and the reproducibility of efflux assays. This study aimed to compare Pgp expression and efflux activity by measuring Rhodamine123 (Rh123) retention in lymphocytes stored under different conditions, in order to evaluate the potential utility of any of the storing conditions in Pgp functionality. Our results show no change in protein expression of Pgp by confocal studies and Western blotting, nor changes at the mRNA level (qRT-PCR). No differences in Rh123 efflux by Pgp activity assays were found between fresh and frozen lymphocytes after 24 hours of blood extraction, using either of the two Pgp specific inhibitors (VP and PSC833). Different working conditions in the 24 hours post blood extraction do not affect Rh123 efflux. These results allow standardization of Pgp activity measurement in different individuals with different timing of blood sampling and in different geographic areas. _______________

Investigación evaluativa sobre el programa "LÍMITES"

Desde la comunidad científica se ha puesto de manifiesto la necesidad de evaluar los programas de intervención para la mejora de la parentalidad, especialmente en el contexto de la infancia y la adolescencia en riesgo. En esta investigación se presentan los resultados de la evaluación del programa Límites aportando evidencias de eficacia y efectividad, que permiten identificar y describir sus fortalezas, así como también aquellos elementos que pueden ser objeto de mejora. El programa Límites es una propuesta estructurada de intervención preventiva de carácter selectivo, con grupos de familias de jóvenes de entre 14 y 18 años que pasan por el circuito de la justicia juvenil. El programa ha estado funcionando desde 2007 en todo el territorio catalán, y ha incidido en un total de 351 personas y 245 familias. El programa consta de ocho sesiones en grupo en las que se trabajan contenidos relacionados con las relaciones y los vínculos, la comunicación, los conflictos, el establecimiento de normas y la disciplina. Para llevar a cabo la evaluación del programa Límites se ha partido de un diseño cuasi-experimental, pretest-postest aplicado a un grupo programa (GP) y grupo colaborador (GC) y de manera diferida (dos meses después de su finalización) sólo en el GP, para constatar si se han alcanzado los resultados previstos. Cuando se analiza la evolución del GP se observan mejoras en las habilidades de autocontrol de las emociones, el refuerzo positivo y las relaciones familiares. Sin embargo, desde el punto de vista de las familias no se han observado cambios significativos en la mejora de la percepción de auto eficacia del rol parental respecto al GC antes y después del programa. La percepción mayoritaria es que los objetivos que plantea el programa son excesivamente ambiciosos y en consecuencia, difícilmente alcanzables. Los responsables consideran que, en algunos casos, el perfil de familias derivadas al programa no es el más adecuado puesto que presenten problemáticas mucho más agudas a las deseables para este tipo de intervención. Como puntos fuertes, se destaca que el programa es útil a los ojos de los responsables y técnicos porque genera una toma de conciencia del problema y de la necesidad de cambio por parte de las familias. Los participantes dicen transferir a la vida real algunas habilidades aprendidas, especialmente la comunicación y mejora de las relaciones familiares. Los técnicos aplicadores coinciden con la percepción de las familias, a pesar de declarar porcentajes ligeramente inferiores. También es visto como un complemento profesional a su tarea que ayuda a hacer un mejor seguimiento de los casos, y permite seguir trabajando con posterioridad con las familias.