Osmotically-induced genes are controlled by the transcription factor TonEBP in cultured cardiomyocytes.

Changes in cardiac osmolarity occur in myocardial infarction. Osmoregulatory mechanisms may, therefore, play a crucial role in cardiomyocyte survival. Tonicity-responsive enhancer binding protein (TonEBP) is a key transcription factor participating in the adaptation of cells to increases in tonicity. However, it is unknown whether cardiac TonEBP is activated by tonicity. Hypertonicity activated transcriptional activity of TonEBP, increased the amounts of both TonEBP mRNA and protein, and induced both the mRNA and protein of TonEBP target genes (aldose reductase and heat shock protein-70). Hypotonicity decreased the amount of TonEBP protein indicating bidirectional osmoregulation of this transcription factor. Adenoviral expression of a dominant negative TonEBP suppressed the hypertonicity-dependent increase of aldose reductase protein. These results indicated that TonEBP controls osmoregulatory mechanisms in cardiomyocytes.

Adherencia a la guía de manejo de sepsis severa y choque séptico en pacientes mayores de 65 años que ingresan a UCI / Adherence to the guidelines for the management of severe sepsis and septic shock in patients over 65 years of age admitted to the ICU

Introduction: The use of guidelines for goal-oriented resuscitation in patients with severe sepsis and septic shock has a positive impact on multiple organ failure and mortality outcomes. However, in patients over 65, adherence to the guidelines may be less stringent because of considerations of lower functional, cardiac, pulmonary and renal reserve. This study compares adherence to the guidelines and compliance with resuscitation goals by the healthcare staff in populations over and under 65 years of age. Objective: To determine adherence to the guidelines by the healthcare staff in the treatment of severe sepsis and septic shock in the group over 65 years of age, compared with patients under 65. Materials and methods: Observational, analytical, cross-sectional study of patients under and over 65 years of age admitted to the Intensive Care Unit (ICU) after having been diagnosed with severe sepsis and septic shock. Frequency of adherence to the guidelines was determined and a comparison of the different resuscitation goals in relation to adherence to the guidelines was made. Results: Except for adherence to early initiation of antibiotic therapy, there was better adherence to the other goals in the patients under 65 when compared to patients over 65. Adherence to the guidelines declined progressively as the number of goals increased. Conclusions: There was a lower adherence to resuscitation guidelines in the group of patients over 65 with severe sepsis and septic shock when compared with the group of patients under 65.

Introducción: El uso de guías de reanimación por metas en los pacientes con sepsis severa y choque séptico, genera un impacto benéfico en los desenlaces de disfunción orgánica múltiple y de la mortalidad, sin embargo en los pacientes mayores de 65 años, la adherencia a las guías puede ser menor, por la menor reserva funcional, cardíaca, pulmonar y renal. En el presente estudio comparamos la adherencia a la guía y el cumplimiento de las metas de reanimación por parte del personal de salud en la población mayor y menor de 65 años. Objetivo: Determinar la adherencia a las guías para el tratamiento de la sepsis severa y choque séptico por parte del personal de salud en el grupo mayor de 65 años comparado con el grupo de pacientes menores de 65 años.. Materiales y métodos: Estudio observacional analítico de corte transversal de los pacientes menores y mayores de 65 años ingresados a la Unidad de Cuidados Intensivos (UCI), con diagnóstico de sepsis severa y choque séptico, se determinó la frecuencia de adherencia a la guía para cada grupo de edades y se compararon las diferentes metas de reanimación en relación a la adherencia. Resultados: Excepto por la adherencia del inicio temprano de la antibiótico terapia, las demás metas presentaron una mejor adherencia, en el grupo de pacientes menores de 65 años cuando se compararon con los pacientes mayores de 65 años, la adherencia para ambos grupos disminuyo progresivamente a medida que se consideraron mayor número de metas. Conclusiones: Existe una menor adherencia a las guías de reanimación, en el grupo de grupo de pacientes mayores de 65 años con sepsis severa y choque séptico cuando se compara con el grupo de pacientes menores de 65 años.

Copper-binding proteins in human erythrocytes: searching for potential biomarkers of copper over-exposure.

The recognition that copper is essential but also potentially toxic to humans has prompted the search for biomarkers of copper excess. The experimental approach followed here involves the isolation and subsequent characterization of copper-binding molecules (CuBP) from human erythrocytes. Incubation (0-60 min) of freshly obtained erythrocytes in the presence of increasing concentrations of copper (10-50 microM; as 64Cu-histidine) led to time- and concentration-dependent uptake of the radioisotope. A near-maximal incorporation was attained after 20 min, with 45-55% of the radioactivity being recovered in 20,000 x g hemolysate supernatants (S-20). 64CuBP from S-20 were separated by size exclusion and metal-affinity chromatography. Most radioactivity loaded into a Sephadex G-75 column was recovered in association with molecules of MMr greater than 60 KDa (largely accounted for by hemoglobin; Hb). Only negligible amounts of radioactive Cu were associated with metallothionein. With further purification, the higher MMr 64Cu-binding fractions were resolved by Sephadex G-200 into two major peaks. The cpm/microg protein ratios of the first peak (high MMr) were proportional to the concentrations of copper presented to the erythrocytes. The second one contained mostly Hb molecules. Proteins from the first peak were concentrated in an affinity chromatography mini-column, suited to trap CuBP. The higher-affinity CuBP were eluted as a single peak which comprised around 60% of the load. An SDS-PAGE analysis of such peak reveals the presence of three bands, of which two are non-hemoglobin Cu-binding proteins. The latter, whose identity remains to be established, had MMr of approximately 30 and 40 KDa, respectively. Preliminary data indicate that the two bands bind 64Cu within a range of concentrations, relevant to those expected to occur during copper over-exposure conditions.