Caveolae (CAV) constitute a novel subcellular transport vesicle that has received special attention based on its proven and postulated participation in transcytosis, potocytosis, and in cell signaling events. One of the principal components of CAV are caveolin protein isoforms. Here, we have undertaken the immunochemical identification of CAV and the known caveolin isoforms (1alpha, 1beta, 2 and 3) in cultured rat C6 glioma cells. Immunoblot analysis revealed that particulate fractions from rat C6 glioma cells express caveolin-1 and caveolin-2. The relative detergent-insolubility of these caveolin isoforms was also determined by Western blot analysis. Indirect immunofluorescence analysis with caveolin-1 and -2 antibodies revealed staining patterns typical of CAV's known subcellular distribution and localization. For both caveolin isoforms immunocytochemical staining was characterized by intensely fluorescent puncta throughout the cytoplasm and diffuse micropatches at the level of the plasmalemma. Perinuclear staining was also detected, consistent and suggestive of caveolin's localization in the trans Golgi region. The caveolin-1 and -2 immunoreactivity seen in Western blots and immunocytochemically is related to structurally relevant CAV as supported by the isolation of caveolin-enriched membrane complexes using two different methods. Light-density, Triton X-100-insoluble caveolin-1- and caveolin-2-enriched fractions were obtained after fractionation of rat C6 glioma cells and their separation over 5-40% discontinuous sucrose-density gradients. Similar fractions were obtained using a detergent-free, sodium carbonate-based fractionation method. These results further support the localization of CAV and caveolins in glial cells. In addition, they demonstrate that cultured C6 glioma cells can be useful as a model system to study the role of CAV and caveolins in subcellular transport and signal transduction events in glial cells and the brain.
Temporary abdominal closure (TAC) has increasingly been employed in the management of severely injured patients to avoid abdominal compartment syndrome (ACS) and as part of damage control surgery (DCS). Although the use of TAC has received great interest, few data exist describing the morbidity and mortality associated with its use in trauma victims. The main goal of this study is to describe the incidence of surgical complications following the use of TAC as well as to define the mortality associated with this procedure. A retrospective review of patients admitted to a state-designated level 1 trauma center from April 2000 to February 2003 was performed. Inclusion criteria were age >18 years, traumatic injury, and need for exploratory laparotomy and use of TAC. A total of 120 patients were included in the study. The overall mortality of trauma patients requiring TAC was 59.2 per cent. The most common causes of death were acute inflammatory process (50.7%), followed by hypovolemic shock (43.7%). The incidence of surgical complications was 26.6 per cent. Intra-abdominal abscesses were the most frequent surgical complication (10%). After multiple logistic regression analysis, increasing age and a numerically greater initial base deficit were found to be independent predictors of mortality in trauma patients that require TAC.
In this group of patients, growth of bacteria in simultaneous cultures was associated with higher mortality-a factor that may be considered a poor prognostic indicator. This factor requires further studies to explore the relation with survival in critically injured patients.
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