Protein kinase C (PKC) plays a major role in regulating cell growth, transformation, and gene expression; however, identifying phosphorylation events that mediate these responses has been difficult. We expressioncloned a group of PKC-binding proteins and identified a high molecular weight, heat-soluble protein as the major PKC-binding protein in REF52 fibroblasts (Chapline, C., Mousseau, B., Ramsay, K., Duddy, S., Li, Y., Kiley, S. C., and Jaken, S. (1996) J. Biol. Chem. 271, 6417-6422). In this study, we demonstrate that this PKC-binding protein, clone 72, is also a PKC substrate in vitro and in vivo. Using a combination of phosphopeptide mapping, Edman degradation, and electrospray mass spectrometry, serine residues 283, 300, 507, and 515 were identified as the major in vitro PKC phosphorylation sites in clone 72. Phosphorylation state-selective antibodies were raised against phosphopeptides encompassing each of the four phosphorylation sites. These antibodies were used to determine that phorbol esters stimulate phosphorylation of serines 283, 300, 507, and 515 in cultured cells, indicating that clone 72 is directly phosphorylated by PKC in living cells. Phosphorylated clone 72 preferentially accumulates in membrane protrusions and ruffles, indicating that PKC activation and clone 72 phosphorylation are involved in membrane-cytoskeleton remodeling. These data lend further evidence to the model that PKCs directly interact with, phosphorylate, and modify the functions of a group of substrate proteins, STICKs (substrates that interact with C-kinase). Protein kinase C (PKC)1 is a family of phospholipid-dependent protein kinases expressed in all cells and tissues (1). PKC activity has been associated with a variety of growth and pathological defects, indicating that PKCs are involved in regulating fundamental cellular processes. PKC is the major cellular receptor for tumor-promoting phorbol esters, and consequently, PKC activation has been linked to later events in multistage carcinogenesis, i.e. tumor promotion/progression. Phorbol esters and other PKC activators rapidly induce a variety of cellular responses including cell shape changes, cytoskeletal remodeling, decreased cell-cell communication, and increased exocytosis. The diversity of the observed responses has made it complicated to focus on particular phosphorylation events that mediate changes in biological activity. Identifying target proteins and determining how PKC phosphorylation alters their activities are keys to understanding the role of PKC in cell regulation.Several approaches have been used to identify PKC-binding proteins. PICKs (proteins that interact with C-kinase) are PKC substrates identified by yeast two-hybrid screening (2). RACKs (receptors for activated C-kinase) are non-substrate proteins that bind to catalytically active PKC. These may target active PKCs to the vicinity of appropriate substrate proteins (3). Several years ago, we began using a PKC overlay assay to identify PKC-interacting proteins (4 -6). We found that many of the bind...

Analysis 1.1. Comparison 1 Cognitive-behavioural coping skills training (CBCST) versus twelve-step facilitation (TSF) programme, Outcome 1 Alcohol abstinence as number achieving 3 or more weeks of consecutive alcohol abstinence during treatment.

Background Although it is accepted that providing medical students with opportunities to engage in research activity is beneficial, little data has been collated on how medical degree curricula may address this issue. This review aims to address this knowledge gap by conducting a scoping review examining curriculum initiatives that seek to enhance research experience for medical students. Methods This review looks to specifically look at ’doing research’ as defined by the MEDINE 2 consensus rather than ‘using research’ for the bachelor component of the Bologna Cycle. The framework developed by Arksey & O’Malley was utilised and a consultation with stakeholders was incorporated to clarify and enhance the framework. Results A total of 120 articles were included in this scoping review; 26 related to intercalated degree options and 94 to non-intercalated degree options. Research initiatives from the United States were most common (53/120 articles). For non-intercalated research options, mandatory and elective research projects predominated. The included studies were heterogeneous in their methodology. The main outcomes reported were student research output, description of curriculum initiative(s) and self-reported research skills acquisition. For intercalated degree options, the three main findings were descriptions of more ‘novel’ intercalated degree options than the traditional BSc, student perspectives on intercalating and the effect of intercalating on medical student performance and careers. Conclusions There are several options available to faculty involved in planning medical degree programmes but further research is needed to determine whether research activity should be optional or mandatory. For now, flexibility is probably appropriate depending on a medical school’s resources, curriculum, educational culture and population needs.

Background: Addiction is a context-specific but common and devastating condition. Though several evidence-based treatments are available, many of them remain under-utilized, among others due to the lack of adequate training in addiction medicine (AM). AM Training needs may differ across countries because of difference in discipline and level of prior AM training or contextual factors like epidemiology and availability of treatment. For appropriate testing of training needs, reliability and validity are key issues. The aim of this study was to evaluate the psychometric properties of the AM-TNA Scale: an instrument specifically designed to develop the competence-based curriculum of the Indonesian AM course. Methods: In a cross-sectional study in Indonesia, Ireland, Lithuania, and the Netherlands the AM-TNA was distributed among a convenience sample of health professionals working in addiction care in The Netherlands, Lithuania, Indonesia, and General Practitioners in-training in Ireland. 428 Respondents completed the AM-TNA scale. To assess the factor structure, we used explorative factor analysis. Reliability was tested using Cronbach's Alpha, ANOVA determined the discriminative validity. Results: Validity: factor analysis revealed a twofactor structure: One on providing direct patient treatment and care (Factor 1: clinical) and one factor on facilitating/supporting direct patient treatment and care (Factor 2: non-clinical) AM competencies and a cumulative 76% explained variance. Reliability: Factor 1 α=.983 and Factor 2: α=.956), while overall reliability was (α=.986). The AM-TNA was able to differentiate training needs across groups of AM professionals on all 30 addiction medicine competencies (P=.001). Conclusions: In our study, the AM-TNA scale had a strong two-factor structure and proofed to be a reliable and valid instrument. The next step should be the testing external validity, strengthening discriminant validity and assessing the re-test effect and measuring changes over time.