[Development of the Saxon Health Target "Active aging - aging in health, autonomy, and participation"]. / Entwicklung des Sächsischen Gesundheitsziels "Aktives Altern - Altern in Gesundheit, Autonomie und Mitverantwortlichkeit."

In Saxony, the consequences of demographic aging are observable already today. To manage the implications on the health sector, the Saxon Health Targets Steering Committee decided in March 2008 to develop a health target "Active Aging - Aging in Health, Autonomy, and Participation". Target development was based on a 7-level approach (fields of action, main goals, target areas, targets, strategies, intervention measures, indicators for evaluation). A quantitative content analysis was used to reveal 10 potential relevant fields of action, three of which were selected for target development. Targets were developed by 53 stakeholders in multiprofessional working groups. Criteria-based analyses were performed to assure appropriate scientific evidence and feasibility of targets and intervention measures. Over a period of 9 months, 24 targets were defined referring to the main goals "needs-based health care structures", "multiprofessional qualification", "self-rated health" and "intergenerational solidarity". Thirteen targets were developed into recommendations for specific intervention measures. Most of the proposed interventions aim to modify health-related structures or psychosocial determinants of health in the elderly. The best recommendations for intervention measures shall be implemented in cooperation with interested decision-makers.

4'-Amino-benzamido-taurocholic acid selectively solubilizes glycosyl-phosphatidylinositol-anchored membrane proteins and improves lipolytic cleavage of their membrane anchors by specific phospholipases.

Glycosyl-phosphatidylinositol-anchored membrane proteins (GPI-proteins) are normally identified either by cleavage of the lipid anchor using (glycosyl)phosphatidylinositol-specific phospholipases C or D (GPI-PLs) or by metabolic labeling of the lipid moiety with specific building blocks. Therefore, methods for discrimination between transmembrane proteins and GPI-proteins on the basis of their physicochemical properties are desirable. Here we are presenting a selective extraction method for typical well-characterized mammalian GPI-proteins, e.g., acetylcholine esterase, alkaline phosphatase, 5'-nucleotidase, and lipoprotein lipase, using a derivative of taurocholate. The results were compared to those obtained with well-characterized transmembrane proteins, e.g., insulin receptor and hydroxymethyl glutaryl coenzyme A-reductase, glucose transporters, or aminopeptidase M and several commercially available detergents. With regard to total membrane proteins, it was possible to selectively enrich GPI-proteins up to 8- to 14-fold by using concentrations between 0.1 and 0.3% of 4'-NH2-amino-7 beta-benzamido-taurocholic acid (BATC). In addition, the cleavage specificity and efficiency of (G)PI-PLs were increased in the presence of identical concentrations of BATC compared to commonly used detergents, e.g., Nonidet P-40. Therefore, the present study shows that the use of BATC facilitates the identification of glycosyl-phosphatidylinositol-anchored membrane proteins.

Charge heterogeneity of insulin fusion proteins expressed in Escherichia coli is not due to proteolytic degradation.

Evaluation of the yield of expression of exogeneous protein in transformed Escherichia coli cells by means of one-dimensional SDS-PAGE often leads to overestimation and miscalculation. For example, it is possible that proteins of similar size comigrate and thus mask the overexpressed product band. Therefore, two-dimensional electrophoresis was used to analyze two types of recombinant fusion proteins, i.e., a beta-galactosidase insulin fusion protein and a interleukin II insulin fusion protein, directly after fermentation. We found that production scale expression products show charge and size heterogeneity. The heterogeneous protein spots were characterized by subsequent blotting onto Immobilon membrane and by N-terminal sequencing. Some of the separated spots were either N-terminally blocked or already degraded to some extent. The integrity of the actual product component of the fusion protein was examined with a C-terminus-specific antibody and by Western blot analysis of the 2D gels.