Chicken oviductal ecto-ATP-diphosphohydrolase. Purification and characterization

An ecto-ATP diphosphohydrolase (ATPDase) was purified to homogeneity from vesiculosomes shed from chicken oviduct. First, the ecto-ATPDase-enriched vesiculosomes were concentrated by filtration, differential centrifugation, and exclusion chromatography. Next, the nonionic detergent, Nonidet P-40, was used to extract the ecto-ATPDase from vesiculosomal membranes, and the solubilized enzyme was further purified by ion exchange (DEAE-Bio-Gel) and lentil-lectin-Sepharose 4B chromatography. In the final stage, immunoaffinity chromatography was utilized to obtain purified ecto-ATPDase. More than 25,000-fold purification was achieved. Specific activity of the purified enzyme was greater than 800 micronol/min/mg of protein with MgATP as the substrate, the highest ever reported for an ATPDase. The enzyme also hydrolyzed other nucleoside triphosphates in the presence of magnesium at similar rates and CaATP and MgADP at lower rates. The molecular mass of the purified glycoprotein was 80 kDa as determined by SDS-polyacrylamide gel electrophoresis and Western blot analysis. Based on its enzymatic properties, the relationship of the chicken oviduct ecto-ATPDase with other reported ATPDases and ecto-ATPases is discussed.     

Decisive arrival law parameters and a general finite capacity queueing problem

This paper addresses the problem of finding the parameters of the arrival law which most significantly influence expected occupation and loss of a finite capacity queue. The input process is supposed to be ergodic and wide sense stationary. We show that it is mostly possible to fit an MMPP(2) to the decisive parameters of observational data. Numerical examples illustrate the importance of the decisive parameters, called key parameters, and also show the accuracy of the proposed fitting procedure. Finally, in the appendix we present the solution of the finite capacity queueing problem with Special Semi Markov Process (SSMP) arrivals and a general service strategy.     

The branch site adenosine is recognized differently for the two steps of pre-mRNA splicing

The functional groups responsible for branch site identity in the two steps of pre-mRNA splicing as well as for spliceosome assembly were tested by incorporation of site-specific modifications at the branch site of a pre-mRNA. These results show that recognition of the adenosine occurs early in complex formation and that the branch site adenosine is recognized differently before the first step and for the second step. Further, direct UV cross-linking with these modified RNAs was used to identify a factor whose interaction was dependent upon the identity of the branch site nucleotide.     

A cross validation study of deep brain stimulation targeting: from experts to atlas-based, segmentation-based and automatic registration algorithms

Validation of image registration algorithms is a difficult task and open-ended problem, usually application-dependent. In this paper, we focus on deep brain stimulation (DBS) targeting for the treatment of movement disorders like Parkinson's disease and essential tremor. DBS involves implantation of an electrode deep inside the brain to electrically stimulate specific areas shutting down the disease's symptoms. The subthalamic nucleus (STN) has turned out to be the optimal target for this kind of surgery. Unfortunately, the STN is in general not clearly distinguishable in common medical imaging modalities. Usual techniques to infer its location are the use of anatomical atlases and visible surrounding landmarks. Surgeons have to adjust the electrode intraoperatively using electrophysiological recordings and macrostimulation tests. We constructed a ground truth derived from specific patients whose STNs are clearly visible on magnetic resonance (MR) T2-weighted images. A patient is chosen as atlas both for the right and left sides. Then, by registering each patient with the atlas using different methods, several estimations of the STN location are obtained. Two studies are driven using our proposed validation scheme. First, a comparison between different atlas-based and nonrigid registration algorithms with a evaluation of their performance and usability to locate the STN automatically. Second, a study of which visible surrounding structures influence the STN location. The two studies are cross validated between them and against expert's variability. Using this scheme, we evaluated the expert's ability against the estimation error provided by the tested algorithms and we demonstrated that automatic STN targeting is possible and as accurate as the expert-driven techniques currently used. We also show which structures have to be taken into account to accurately estimate the STN location.     

Pocket pagers in lots of one CIM

The Fusion program, a method for agile, flexible computer integrated manufacturing (CIM) at Motorola's Paging Products Group, is discussed. Fusion's CIM and automated assembly system can manufacture a wide variety of different products on the same production line. The development of the Fusion program and how it differs from its predecessor, the Bandit program, are described     

SOAT1: A Suitable Target for Therapy in High-Grade Astrocytic Glioma?

Targeting molecular alterations as an effective treatment for isocitrate dehydrogenase-wildtype glioblastoma (GBM) patients has not yet been established. Sterol-O-Acyl Transferase 1 (SOAT1), a key enzyme in the conversion of endoplasmic reticulum cholesterol to esters for storage in lipid droplets (LD), serves as a target for the orphan drug mitotane to treat adrenocortical carcinoma. Inhibition of SOAT1 also suppresses GBM growth. Here, we refined SOAT1-expression in GBM and IDH-mutant astrocytoma, CNS WHO grade 4 (HGA), and assessed the distribution of LD in these tumors. Twenty-seven GBM and three HGA specimens were evaluated by multiple GFAP, Iba1, IDH1 R132H, and SOAT1 immunofluorescence labeling as well as Oil Red O staining. To a small extent SOAT1 was expressed by tumor cells in both tumor entities. In contrast, strong expression was observed in glioma-associated macrophages. Triple immunofluorescence labeling revealed, for the first time, evidence for SOAT1 colocalization with Iba1 and IDH1 R132H, respectively. Furthermore, a notable difference in the amount of LD between GBM and HGA was observed. Therefore, SOAT1 suppression might be a therapeutic option to target GBM and HGA growth and invasiveness. In addition, the high expression in cells related to neuroinflammation could be beneficial for a concomitant suppression of protumoral microglia/macrophages.     

Verfahrenstechnische und wirtschaftliche Vorteile der Kochextrusion im Vergleich zu herkömmlichen Verfahren der Lebensmittel-Industrie

Economic and process engineering advantages of extrusion cooking in comparison with conventional processes in the food industry . Extrusion cooking is introduced as a modern high-temperature short-time process. Possible process steps in extruder machine systems, such as continuous conveying, mixing, homogenizing and reactions, mechanical and thermal energy dissipation for plasticizing and modifying biopolymers, are explained. This process creates new products with completely new textures, for example in the snack and breakfast cereal sector, while on the other hand the extrusion process competes with long-standing conventional processes. The latter work slowly with low pressures, low temperatures and energy dissipation with high water contents in large batch-operated machines. The quality of extruder-cooked products depends on the extrusion system. The self-cleaning corotating twin-screw extruder with its narrow residence time spectrum is the optimum system. However, the counter-rotating intermeshing machine has certain advantages for products of low viscosity. The cooking time at high temperature is a matter of seconds, which serves to maintain the properties of the ingredients and active substances, while ensuring fast destruction of microorganisms. The end-products have a long shelf-life on account of their low process moisture content. Continuous extrusion cooking has economic advantages mainly because it replaces many batch processes and because extrusion is carried out entirely or almost entirely with the final moisture content, thus avoiding the necessity to evaporate huge quantities of water.     

Toxicity of fatty ozonides and peroxides

Studies on the acute toxicity of the ozonides and hydroperoxides of methyl linoleate are reported. High purity preparations of these compounds were injected intravenously or administered orally to adult male rats. The lethal dose by iv injection of these compounds was virtually the same −0.07 mmol/100 g body wt. No deaths were caused in a 24 hr period by single oral dosages of these compounds of ca. 10-fold that causing death by the iv route. The major effect of these compounds was on the lungs. The lungs became enlarged from edema and accumulation of fluid, and the animals died of lung congestion and injury similar to the effects of ozone toxicity. There was no destruction of vitamin E in the tissues of animals given lethal dosages of ozonides or peroxides intravenously, but significant changes occurred in fatty acid composition of the lipids of the serum and lung. Arachidonic acid increased at the expense of linoleic and oleic acids in these tissues. Only small amounts of peroxidic and TBA positive substances were detected in lung and serum, indicating that the injected ozonides and hydroperoxides were destroyed in the tissues.     

New carbon- and sulfur-based ligands in catalysis

Homogeneous catalysis is a field of research that has gained central importance in both organic and inorganic chemistry and the use of well-defined ligand systems in the synthesis of transition metal complexes has had an enormous impact on the development of such catalysts. Neutral, two-electron donor ligands based on phosphorous and nitrogen have been tremendously successful as ancillary entities for late-transition metal (LTM) catalysts, whereas ligands based on anionic nitrogen, oxygen and the cyclopentadienyl motif (Cp(-)) have propelled early-transition metal (ETM) catalysis forward. We believe that expanding the ligand families capable of acting as successful entities in metal-mediated reactivity and catalysis is crucial for future discoveries in this field. Research in our group therefore tries to identify new non-chiral and chiral ligands for late-transition metal chemistry that are based on neutral, two-electron carbon and sulfur donor atoms. In particular, we have until now focused on the development of modular, monodentate N-heterocyclic carbene ligands (NHCs) that can serve as a basis for the development of chiral ligand frameworks for the application to asymmetric catalytic transformations. In the second major research project developed over the last six years, we have started an investigation on the use of chelating sulfoxide-based ligands in asymmetric late transition-metal based catalysis.