Aurora Kinase A Is Involved in Controlling the Localization of Aquaporin-2 in Renal Principal Cells

The cAMP-dependent aquaporin-2 (AQP2) redistribution from intracellular vesicles into the plasma membrane of renal collecting duct principal cells induces water reabsorption and fine-tunes body water homeostasis. However, the mechanisms controlling the localization of AQP2 are not understood in detail. Using immortalized mouse medullary collecting duct (MCD4) and primary rat inner medullary collecting duct (IMCD) cells as model systems, we here discovered a key regulatory role of Aurora kinase A (AURKA) in the control of AQP2. The AURKA-selective inhibitor Aurora-A inhibitor I and novel derivatives as well as a structurally different inhibitor, Alisertib, prevented the cAMP-induced redistribution of AQP2. Aurora-A inhibitor I led to a depolymerization of actin stress fibers, which serve as tracks for the translocation of AQP2-bearing vesicles to the plasma membrane. The phosphorylation of cofilin-1 (CFL1) inactivates the actin-depolymerizing function of CFL1. Aurora-A inhibitor I decreased the CFL1 phosphorylation, accounting for the removal of the actin stress fibers and the inhibition of the redistribution of AQP2. Surprisingly, Alisertib caused an increase in actin stress fibers and did not affect CFL1 phosphorylation, indicating that AURKA exerts its control over AQP2 through different mechanisms. An involvement of AURKA and CFL1 in the control of the localization of AQP2 was hitherto unknown.     

Surface Acoustic Wave (SAW) Biosensor for Rapid and Label-Free Detection of Penicillin G in Milk

Surface acoustic wave (SAW) biosensors were introduced for rapid and label-free detection of penicillin G in milk. This is the first time that the use of SAW biosensors for antibiotic detection is reported. Penicillin G belongs to the β-lactam antibiotics which are commonly used in human and veterinary medicine. Particularly because of the latter, the detection of antibiotics in foodstuffs of animal origin is essential. Current methods for specific antibiotic detection often require complex laboratory equipment and procedures. SAW biosensors, however, offer rapid detection of analyte concentrations with a minimum of experimental effort. They use label-free acoustic (gravimetric) detection. Owing to the small mass of antibiotics, detection via binding inhibition assay was preferred to direct detection in this work. Samples containing penicillin G were preincubated with the corresponding antibody, and SAW biosensor surfaces were coated with penicillin G epitopes. The antibody in the sample bound to the biosensor surface, unless it was inhibited by penicillin G. The assay allowed the detection of 2 ng/ml penicillin G in buffer and 2.2 ng/ml in low-fat milk. This is below the maximum residue limit of 4 ng/ml given by the European Commission for penicillin G and other β-lactam residues in food.     

Einchecken in der Business Class!

Praxis&Wissen

Einchecken in der Business Class! Lage- und Qualit?tsprüfung bei Zuführsystemen     

Carotenoids from tomatoes inhibit heterocyclic amine formation

Tomatoes are a rich source of antioxidants and they are commonly used for meat cooking in Mediterranean countries. The effects of tomato antioxidants on the formation of carcinogenic/mutagenic heterocyclic amines were investigated. A liquid model system containing as precursors creatinine, glucose and glycine in molar concentrations comparable to those present in bovine meat (chemical model system) was employed. A freeze-dried bovine meat juice (meat juice model system) was also used for some experiments. In both model systems, an inhibiting effect of tomato carotenoid fraction on the formation of imidazoquinolines (IQx, MeIQx and DiMeIQx) was observed. Using carotenoid extract at a concentration of 1000 ppm, inhibitions of 36% and 11% of IQx and MeIQx formation respectively in the chemical system and of 13% of MeIQx and of 5% of 4,8-DiMeIQx in the meat juice system was observed. The effect of the main tomato flavonoid, quercetin, was investigated using the meat juice system. Quercetin gave an inhibition of MeIQx formation between 9% and 57% with a maximum effect of 67% at 10 ppm.     

SWOF - an open framework for shared workspaces to support different cooperation tasks

Computer supported collaborative work (CSCW) allows people to cooperate by computers from different places and at different times. To enable an easier integration of such collaborative components into web-based communities and portals, a Shared Workspace Open Framework (SWOF) was developed. This framework provides the basic features of shared workspaces and can be customized to different cooperation cases. High usability is an important aspect of the implementation. To achieve these aims SWOF focuses on an information space with more task-suited item-types that can help to pre-structure the information. Thus, on the one hand the system can help the users to write down the needed information in a consistent way and, on the other, could reduce the arguments between the group members on how to structure their workspace. As a use case for SWOF the development of a Web-based portal for the community of man-machine interaction was chosen. In the project MMI-Interaktiv, a portal is built with a SWOF-based shared workspace component. An evaluation for this use case is presented.     

Heterocyclic amines in some Swedish cooked foods industrially prepared or from fast food outlets and restaurants

Accurate assessment of human intake of mutagenic/carcinogenic heterocyclic amines (HAs) is necessary for epidemiological studies and future risk assessment. Using questionnaires, the frequency of consumption of specific dishes can be obtained at an individual level and linked to analyzed concentrations of different compounds in corresponding dishes. Some typical Swedish cooked meat dishes, hamburgers and kebab, industrially prepared or from fast food outlets and restaurants, were analyzed regarding their content of 11 different HAs. The amount of each of these compounds was below 0.1 ng/g cooked weight in most of the industrially prepared products. The total amount of HAs was highest in the kebab samples. The intake of HAs from 200 g of the dishes was estimated to range from not detectable levels to 0.6 microg. The results of the present study indicate that the content of HAs in a specific dish may vary with origin, and that the concentrations of HAs in commercial fried meat products are generally low, although some of these food items may contain elevated amounts.     

Determining the performance of website-based relationship marketing

Company websites are an important instrument for relationship marketing activities. We present a methodological framework that aligns website performance assessment and marketing intelligence for evaluating the performance of relationship marketing activities. In this context, we develop an extended web mining approach that integrates managerial perspectives in the analyst’s investigation of the customer-website interaction based on historical clickstream data.This approach enables quantification of the moderating effect of a website’s structure and content regarding website-based relationship marketing. The applicability of our approach is demonstrated by clickstream data of 477,471 visitor sessions on a software developer’s website. The results provide detailed insights into the usage behavior on a website and the mechanisms to enhance e-commerce efficiency via website optimization.     

Several ways to produce porous carbon monoliths by template assisted routes

Several ways to produce microporous and mesoporous carbon monoliths by template assisted synthesis routes are described. Pressed zeolite (HY) and silica powder tablets have been used as templates. Sucrose acts as a carbon precursor and as a binder between the carbon particles as well. In the case of zeolite template, the carbon porosity could be increased by additional loading of the template with ethene. Due to the balance between mechanic stability and porosity of the carbon monolith, an optimal amount of sucrose is required. The final products were characterized by low temperature nitrogen adsorption.