Pharmakogenetik der oralen Antikoagulation mit Cumarinen

The recent identification of VKORC1 has made important contributions to our understanding of the vitamin K cycle. The VKORC1 enzyme was shown to be the molecular target of coumarin drugs. Mutations and polymorphisms in coding and noncoding regions of the VKORC1 gene have been shown to cause both a partial to total coumarin resistance and coumarin sensitivity. Availability of molecular diagnostics (VKORC1, CYP2C9) and drug monitoring by HCPLC (determination of coumarin, vitamin K, and vitamin K epoxide levels) is helpful for detecting hereditary and acquired factors influencing coumarin therapy. In the future, these tools may be instrumental in designing individualized oral anticoagulation therapy regimens.     

Evaluation of the efficiency of measures for sulphidic mine waste mitigation

To control the environmentally detrimental impact of acid rock drainage, two different countermeasures, layers of acid-buffering materials and sodium dodecyl sulphate addition, were tested for their efficiency in laboratory percolation experiments. In the experiment with a layer of calcium bentonite, only the iron output was reduced. The experiments with layers of concrete grains demonstrated a decrease of the microbial activity as well as a precipitation of heavy-metal ions, whereas the cell numbers did not decrease. Furthermore, finely grained concrete (1–5 mm) formed a water-tight hardpan (self-sealing layer). In the experiment with 1 mM sodium dodecyl sulphate, all the microorganisms were killed and hence metal sulphide dissolution was stopped. With 0.1 mM sodium dodecyl sulphate only a short, transient inhibition of leaching was achieved. The bacteria remained alive. Received: 16 February 1998 / Accepted: 23 February 1998     

Transposition-mediated transcriptional overexpression as a mechanism of insecticide resistance

It has been proposed that amplification of genes for esterase that provide resistance to insecticides may originate from transposition events. To test this hypothesis, we have constructed a minigene coding for a soluble acetylcholinesterase under the control of a nontissue-specific promoter (hsp70). When introduced into Drosophila, the gene is expressed in all tissues and the extra acetylcholinesterase produced confers a low level of insecticide resistance (twofold). The minigene was mobilized by crossing the initial transformant with a strain providing a source of P-element transposase. After 34 generations of exposure to the organophosphate parathion, we obtained a strain with a higher resistance (fivefold). This strain had only one extra Ace gene, which overexpressed acetylcholinesterase. Thus, following transposition, resistance resulted from the overexpression of a single copy of the gene and not from gene amplification. Received: 9 August 1996 / Accepted: 27 May 1997     

Human acetylcholinesterase. Immunochemical studies with monoclonal antibodies

Monoclonal antibodies were used to investigate the immunochemistry of human erythrocyte acetylcholinesterase (acetylcholine acetylhydrolase, EC 3.1.1.7). A series of experiments on the sedimentation velocity and Stokes radius of acetylcholinesterase and its immune complexes indicated that each antibody recognized a single high-affinity binding site (epitope) on the monomeric enzyme. Further analysis suggested that the antibody-binding sites were replicated on multimeric enzyme forms but were subject to steric hindrance between nearby IgG molecules or adjacent enzyme subunits. The cellular localization of the epitopes was studied by measuring the binding of monoclonal antibodies to the cholinesterase of intact erythrocytes. The results implied that most of the epitopes are exposed to the external media. However, one antibody failed to bind to intact cells, despite a relatively high affinity for detergent-solubilized antigen, possibly because its epitope is buried in the lipid bilayer.     

IgG production kinetics in serum free media

Summary A murine hybridoma (455) was cultured in four different serum free media formulations, and a newborn calf serum supplemented medium was used as a basis of comparison. The serum supplemented medium supports a higher cell growth rate and results in a higher IgG titer. However, the antibody secretion rate on a per cell basis is higher in the serum free media, indicating that serum could be inhibitory to antibody secretion. The results identify the possibility of a least eliminating serum during the monoclonal antibody production phase.     

Respiration rates of some greenhouse cultivated tropical and subtropical species

A comparative study of dark respiration and effects of high and low temperature on dark respiration in five greenhouse cultivated tropical and sub-tropical species was made. The respiration rates determined manometrically were low in all species. Respiration rates increased with increasing temperature. Low temperature treatment accelerated the rate of respiration at 25 °G except in two species. An attempt has been made to determine the critical and optimum temperature for these species. The respiratory behaviour of these species has been discussed with respect to their original habitats and prevailing environmental conditions.     

Involvement of the galactosyl-1-phosphate transferase encoded by the Salmonella enterica rfbP gene in O-antigen subunit processing.

rfbT of Salmonella enterica LT2 was previously thought, together with rfaL, to be involved in the ligation of polymerized O antigen to core-lipid A, and three mutants were known. We report the mapping of the mutations to rfbP, the galactosyl-1-phosphate transferase gene, which is now shown to encode a bifunctional protein. The mutations which have the former rfbT phenotype are referred to as rfbP(T). We also show that rfbP(T) mutants are not blocked in the ligation step as previously believed but in an earlier step, possibly in flipping the O-antigen subunit on undecaprenyl pyrophosphate from the cytoplasmic to periplasmic face of the cytoplasmic membrane.