Identification of a cyclase-associated protein (CAP) homologue in Dictyostelium discoideum and characterization of its interaction with actin

In search for novel actin binding proteins in Dictyostelium discoideum we have isolated a cDNA clone coding for a protein of approximately 50 kDa that is highly homologous to the class of adenylyl cyclase-associated proteins (CAP). In Saccharomyces cerevisiae the amino-terminal part of CAP is involved in the regulation of the adenylyl cyclase whereas the loss of the carboxyl-terminal domain results in morphological and nutritional defects. To study the interaction of Dictyostelium CAP with actin, the complete protein and its amino-terminal and carboxyl-terminal domains were expressed in Escherichia coli and used in actin binding assays. CAP sequestered actin in a Ca2+ independent way. This activity was localized to the carboxyl-terminal domain. CAP and its carboxyl-terminal domain led to a fluorescence enhancement of pyrene-labeled G-actin up to 50% indicating a direct interaction, whereas the amino-terminal domain did not enhance. In polymerization as well as in viscometric assays the ability of the carboxyl-terminal domain to sequester actin and to prevent F-actin formation was approximately two times higher than that of intact CAP. The sequestering activity of full length CAP could be inhibited by phosphatidylinositol 4,5-bisphosphate (PIP2), whereas the activity of the carboxyl-terminal domain alone was not influenced, suggesting that the amino-terminal half of the protein is required for the PIP2 modulation of the CAP function. In profilin-minus cells the CAP concentration is increased by approximately 73%, indicating that CAP may compensate some profilin functions in vivo. In migrating D. discoideum cells CAP was enriched at anterior and posterior plasma membrane regions. Only a weak staining of the cytoplasm was observed. In chemotactically stimulated cells the protein was very prominent in leading fronts. The data suggest an involvement of D. discoideum CAP in microfilament reorganization near the plasma membrane in a PIP2-regulated manner.     

Plasma Metabolomics Analysis of Polyvinyl Chloride Workers Identifies Altered Processes and Candidate Biomarkers for Hepatic Hemangiosarcoma and Its Development

Background: High-level occupational vinyl chloride (VC) exposures have been associated with hepatic hemangiosarcoma, which typically develops following a long latency period. Although VC is genotoxic, a more comprehensive mode of action has not been determined and diagnostic biomarkers have not been established. The purpose of this study is to address these knowledge gaps through plasma metabolomics. Methods: Plasma samples from polyvinyl chloride polymerization workers who developed hemangiosarcoma (cases, n = 15) and VC exposure-matched controls (n = 17) underwent metabolomic analysis. Random forest and bioinformatic analyses were performed. Results: Cases and controls had similar demographics and routine liver biochemistries. Mass spectroscopy identified 606 known metabolites. Random forest analysis had an 82% predictive accuracy for group classification. 60 metabolites were significantly increased and 44 were decreased vs. controls. Taurocholate, bradykinin and fibrin degradation product 2 were up-regulated by greater than 80-fold. The naturally occurring anti-angiogenic phenol, 4-hydroxybenzyl alcohol, was down-regulated 5-fold. Top affected ontologies involved: (i) metabolism of bile acids, taurine, cholesterol, fatty acids and amino acids; (ii) inflammation and oxidative stress; and (iii) nicotinic cholinergic signaling. Conclusions: The plasma metabolome was differentially regulated in polyvinyl chloride workers who developed hepatic hemangiosarcoma. Ontologies potentially involved in hemangiosarcoma pathogenesis and candidate biomarkers were identified.     

Hydroisomerization of Renewable and Fossil n-Alkanes over Bifunctional Dealuminated ZSM-5 Catalysts

The hydroisomerization of two long-chain n-alkane mixtures was investigated over bifunctional Pt/H-ZSM-5 catalysts before and after dealumination of preshaped zeolite/binder pellets. The hydroisomerization over the dealuminated catalysts leads to more isomers and less cracking products. Consequently, higher ratios of multi- to mono-branched isomerization products are formed as expected for large- rather than medium-pore zeolites. This indicates a higher availability of space in the vicinity of the active sites and provides an attractive route to make medium-pore zeolites suitable for upgrading higher boiling hydrocarbon feeds.     

Personalisierte Medizin als Orphanisierung: rechtliche und ethische Fragen

Definition of the problem The development of personalized medicine is currently a popular topic. Particularly the personalized pharmacotherapy is a matter of relative importance due to pharmacological and molecular–genetic developments. For this reason it is required to examine the implications of personalized pharmacotherapy to statutory health insurance and health care of patients. Arguments In this context the question of orphanization arises: If medical drugs of personalized medicine could be regularly indicated as orphan drugs, i.e., as drugs to medicate orphan diseases, it would not be necessary according to the “Arzneimittelneuordnungsgesetz” (AMNOG) to prove an auxiliary value within the assessment of the “Gemeinsamer Bundesausschuss” (GBA). Crucial to the drug’s indication as an orphan drug is among other things the rate of prevalence of the patient group, whose disease should be medicated with the drug. In this connection, there is importance attached to the stratification of patient groups within personalized medicine. Conclusion An orphanization causes consequences in terms of prices to the statutory health insurance due to the lack of knowledge about an auxiliary value. For patients, the implications must be considered in a more differentiated manner.     

Beam purification by photodetachment (invited)

Ion beam purity is of crucial importance to many basic and applied studies in nuclear science. Selective photodetachment has been proposed to suppress unwanted species in negative ion beams while preserving the intensity of the species of interest. A highly efficient technique based on photodetachment in a gas-filled radio frequency quadrupole ion cooler has been demonstrated. In off-line experiments with stable ions, up to 10(4) times suppression of the isobar contaminants in a number of interesting radioactive negative ion beams has been demonstrated. For selected species, this technique promises new experimental possibilities in studies on exotic nuclei, accelerator mass spectrometry, and fundamental properties of negative atomic and molecular ions.     

Clotrimazole-loaded Eudragit® RS100 nanocapsules: Preparation,characterization and in vitro evaluation of antifungal activity against Candida species

Clotrimazole is a common choice for the treatment of vulvovaginal infections, but its low solubility and some side effects pose a challenge to its application. This work evaluated the feasibility to formulate clotrimazole-loaded cationic nanocapsules using Eudragit® RS100 and medium chain triglycerides as polymer and oily core, respectively, by the method of interfacial deposition of a preformed polymer. The physicochemical characteristics of nanocapsule formulations were evaluated at 0 day and 60 days after preparation. Particle size, zeta potential, polydispersity index, pH and drug content were stable during this period. In addition, nanocapsules were able to protect clotrimazole from photodegradation under UV radiation. By the dialysis bag diffusion technique, the nanosized formulations showed prolonged release of clotrimazole by anomalous transport and first order kinetics. A microbiological study was carried out by the microdilution method and showed that nanocapsules (mean size: 144 nm; zeta potential: + 12 mV) maintained the antifungal activity of clotrimazole against Candida albicans and Candida glabrata strains susceptible and resistant to fluconazole.     

An empirical investigation on how portfolio risk management influences project portfolio success

Project risk management is recognized as essential in order to cope with the challenges arising from the environment. Literature suggests a portfolio-wide perspective for managing risks in project portfolios. However, research on risk management and its success in a project portfolio context is scarce. This study examines how portfolio risk management influences project portfolio success. Using a sample of 176 firms, this study provides evidence that portfolio risk identification, the formalization of the portfolio risk management process, and risk management culture directly influence risk transparency, whereas risk prevention, risk monitoring, and the integration of risk management into project portfolio management are directly connected to risk coping capacity. The findings also suggest that both risk transparency and risk coping capacity have a direct impact on project portfolio success. However, the results did not confirm the hypothesis that risk transparency and risk coping capacity have a complementary effect on success. Implications for scholars and project portfolio managers are discussed.     

Grauwasserreinigung mit einer Sumpfpflanzenmatte unter Praxisbedingungen

There is increasing interest worldwide in the decentralized treatment of gray water. Since ground‐level areas are generally costly, gray‐water treatment can also be carried out using helophyte mats on the roofs of buildings as an alternative. Alongside the water treatment itself, this process also has a positive effect on the indoor climate in the building. In a test carried out under realistic conditions, it was shown that a helophyte mat with a root‐layer depth of 0.1 m and with hydraulic loads per unit up to 15 L m^?2d^?1 is suitable for treating typical gray water from a residential building.     

Interleukin‐4‐Clicked Surfaces Drive M2 Macrophage Polarization

Driving macrophage (M?) polarization into the M2 phenotype provides potential against inflammatory diseases. Interleukin‐4 (IL‐4) promotes polarization into the M2‐M? phenotype, but its systemic use is constrained by dose‐limiting toxicity. Consequently, we developed IL‐4‐decorated surfaces aiming at sustained and localized activity. IL‐4 muteins were generated by genetic code expansion; Lys42 was replaced by unnatural amino acids (uAAs). Both muteins showed cell‐stimulation ability and binding affinity to IL4Rα similar to those of wt‐IL‐4. Copper‐catalyzed (CuAAC) and copper‐free strain‐promoted (SPAAC) 1,3‐dipolar azide–alkyne cycloadditions were used to site‐selectively anchor IL‐4 to agarose surfaces. These surfaces had sustained IL‐4 activity, as demonstrated by TF‐1 cell proliferation and M2, but not M1, polarization of M‐CSF‐generated human M?. The approach provides a blueprint for the engineering of cytokine‐activated surfaces profiled for sustained and spatially controlled activity.