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The determination of low-molecular weight substances (haptens) is demonstrated with a homogeneous time-resolved immunoassay using antibody-induced luminescence quenching. Our novel assay technology uses the newly developed monoclonal antibody (G24-BA9) to quench the luminescence of europium trisbipyridine (EuTBP). We performed a competitive biotin immunoassay including an EuTBP-biotin conjugate, the anti-EuTBP antibody G24-BA9 and streptavidin as assay components. Steric hindrance allows only the binding of either G24-BA9 (to the EuTBP moiety) or streptavidin (to the biotin moiety) to the EuTBP-biotin conjugate. Addition of the analyte biotin resulted in the binding of streptavidin to biotin and a concomitant preferred binding of G24-BA9 to EuTBP-biotin. Since G24-BA9 quenches the luminescence of EuTBP within the conjugate, the luminescence signal could be used to indicate and quantify the presence of free biotin in the system. All experiments were carried out in solution in the presence of 5% serum demonstrating the possibility of using our novel assay for a very fast determination of low molecular weight substances in biological fluids.
In this article, five cases of odontogenous dysfunctions and musculoskeletal complaints are presented. A common finding in all patients of this study was that the presence of joint complaints was related to deficits in the corresponding muscular function. These deficits were determined by manual muscle tests as described by Kendall et al. [Muscles - Testing and Function, ed 4. Baltimore, Williams and Wilkins, 1993] and were eliminated immediately by a neural therapeutic test injection into the disturbed dental region. The therapy provided solely aimed to eliminate the odontogenous dysfunction. No other therapeutic measures were carried out with regard to the patients' respective muscle, tendon, or joint complaints.
Understanding the interactions between the different processes that control the geothermal and fluid flow fields in sedimentary basins is crucial for exploitation of geothermal energy. Numerical models provide predictive and feasible information for a correct assessment of geothermal resources especially in areas where data acquisition is demanding. Here, we present results from numerical efforts to characterize the thermal structure and its interaction with the fluid system for the area of the North East German Basin (NEGB). The relative impact of the different (diffusive and advective) processes affecting the hydrothermal setting of the basin are investigated by means of three- dimensional numerical simulations. Lithospheric-scale numerical models are evaluated to understand the specific thermal signature of the relevant factors influencing the present-day conductive geothermal field in the NEGB. Shallow and deep structural controls on the thermal configuration of the basin are addressed and quantified. Interaction between the resulting thermal field and the active fluid system is investigated by means of three-dimensional simulations of coupled fluid flow and heat transport. Factors influencing stability and reliability of modeling predictions are discussed. The main effort is to build a physically consistent model for the basin which integrates the impacts of thermal gradients on the regional fluid regime and their coupling with the main geological units defining the basin.
Three dimensional modelling of fractured and faulted reservoirs : framework and implementation
(2010)
Modelling of coupled physical processes in fractured and faulted media is a major challenge for the geoscience community. Due to the complexity related to the geometry of real fracture networks and fault systems, modelling studies have been mainly restricted either to two dimensional cases or to simplified orthogonal fracture systems consisting of vertical and horizontal fractures. An approach to generate three dimensional meshes for realistic fault geometries is presented. The method enables representation of faults in an arbitrary incline as two dimensional planes within a three dimensional, stratified porous matrix of a generic geometry. Based on a structural geological model, the method creates three dimensional unstructured tetrahedral meshes. These meshes can be used for finite element and finite volume numerical simulations. A simulation of a coupled fluid flow and heat transport problem for a two layered porous medium cut by two crossing faults is presented to test the reliability of the method.
A monoclonal antibody against the potential tumor suppressor kinase-enhanced protein phosphatase 1 (PP1) inhibitor KEPI (PPP1R14C) was generated and characterized. Human KEPI was expressed in Escherichia coli and used to immunize Balb/c mice. Using hybridoma technology, one clone, G18AF8, was isolated producing antibodies which bound specifically to the KEPI protein in ELISA, immunoblotting and flow cytometry. The antibody was also successfully applied to stain KEPI protein in paraffin sections of human brain. The epitope was mapped using peptide array technology and confirmed as GARVFFQSPR. This corresponds to the N-terminal region of KEPI. Amino acid substitution analysis revealed that two residues, F and Q, are essential for binding. Affinity of binding was determined by competitive ELISA as 1 mu M. In Western blot assays testing G18AF8 antibody on brain samples of several species, reactivity with hamster, rat and chicken samples was found, suggesting a broad homology of this KEPI epitope in vertebrates. This antibody could be used in expression studies at the protein level e.g. in tumor tissues.