CHARACTERIZATION OF CYTOCHROME P-450 MIXED FUNCTION OXIDASE OF RAT COLON MUCOSA

Non-pregnant, female adult rats pretreated with either phenobarbital (PB) or (beta)-naphthoflavone ((beta)NF) through short-course intraperitoneal injections were shown by sodium dithionite-reduced carbon monoxide difference spectroscopy and NADPH-cytochrome c in vitro assay to contain cytochrome P-450 and NADPH-dependent reductase associated with the microsomal fraction of colon mucosa. These two protein components of the mixed function oxidase system were released from the microsomal membrane, resolved from each other, and partially purified by using a combination of techniques including solubilization in nonionic detergent followed by ultracentrifugation, anion exchange and adsorption column chromatographies, native gel electrophoresis, polyethylene glycol fractionation and ultrafiltration.^ In vitro reconstitution assays demonstrated the cytochrome P-450 fraction as the site of substrate and molecular oxygen binding. By the use of immunochemical techniques including radial immunodiffusion, Ouchterlony double diffusion and protein electroblotting, the cytochrome P-450 fraction was shown to contain at least 5 forms of the protein, having molecular weights as determined by SDS gel electrophoresis identical to the corresponding hepatic cytochrome P-450. Estimation of total cytochrome P-450 content confirmed the preferential induction of particular forms in response to the appropriate drug pretreatment.^ The colonic NADPH-dependent reductase was isolated from native gel electrophoresis and second dimensional SDS gel electrophoresis was performed in parallel to that for purified reductase from liver. Comparative electrophoretic mobilities together with immunochemical analysis, as with the cytochrome P-450s, reconstitution assays, and kinetic characterization using artificial electron acceptors, gave conclusive proof of the structural and functional homology between the colon and liver sources of the enzyme.^ Drug metabolism was performed in the reconstituted mixed function oxidase system containing a particular purified liver cytochrome P-450 form or partially pure colon cytochrome P-450 fraction plus colon or liver reductase and synthetic lipid vesicles. The two drugs, benzo{(alpha)}pyrene and benzphetamine, which are most representative of the action of system in liver, lung and kidney, were tested to determine the specificity of the reconstituted system. The kinetics of benzo{(alpha)}pyrene hydroxylation were followed fluorimetrically for 3-hydroxybenzo{(alpha)}pyrene production. . . . (Author's abstract exceeds stipulated maximum length. Discontinued here with permission of author.) UMI ^

Na 50 Nachlass Arthur Schopenhauer - 'Schopenhauer-Archiv', 708 - Nachfrage zur vollzogenen Auszahlung von 450 Thalern

u.a.: Erbschaft; Finanzen; Ohra;

Na 50 Nachlass Arthur Schopenhauer - 'Schopenhauer-Archiv', 450 - Auszug aus John Herrschels "Treatise on Astronomy" in Lardner's Cyclopedia

2 Seiten

Romanian Volach

by Abe Schwartz

Ms.Ff.F.Stoltze 3. 450 - Brief von Friedrich Stoltze an Laura Stoltze

Frankfurter Latern, Produktion

Factors influencing time to HIV disease progression in Romanian children

The purpose of this research was two-fold; to investigate the effect of institutionalization on death and CD4 decline in a cohort of 325 HIV-infected Romanian children, and to investigate the effect of disclosure of the child's own HIV status in this cohort. All children were treated with Kaletra-based highly active antiretroviral therapy, and were followed from November, 2001 through October, 2004. The mean age of the children included in the cohort is 13. The study found that children in biological families were more likely to experience disease progression through either death or CD4 decline than children in institutions (p=0.04). The family home-style institution may prove to be a replicable model for the safe and appropriate care of HIV-infected orphaned and abandoned children and teens. The study also found that children who do not know their own HIV infection status were more likely to experience disease progression through either death or CD4 decline than children who know their HIV diagnosis (p=0.03). This evidence suggests that, in the context of highly active anti retroviral therapy, knowledge of one's own HIV infection status is associated with delayed HIV disease progression. ^

PREPARATION AND CHARACTERIZATION OF FAD DEPENDENT NADPH CYTOCHROME P-450 REDUCTASE (FLAVOPROTEINS, APOFLAVOPROTEIN)

NADPH cytochrome P-450 reductase releases FMN and FAD upon dilution into slightly acidic potassium bromide. The flavins are released with positive cooperativity. Dithiothreitol protects the FAD dependent cytochrome c reductase activity against inactivation by free radicals. Behavior in potassium bromide is sensitive to changes in the pH. High performance hydroxylapatite resolved the FAD dependent reductase from holoreductase. For 96% FAD dependent reductase, the overall yield was 12%.^ High FAD dependence was matched by a low FAD content, with FAD/FMN as low as 0.015. There were three molecules of FMN for every four molecules of reductase. The aporeductase had negligible activity towards cytochrome c, ferricyanide, menadione, dichlorophenolindophenol, nitro blue tetrazolium, oxygen and acetyl pyridine adenine dinucleotide phosphate. A four minute incubation in FAD reconstituted one half to all of the specific activity, per milligram protein, of untreated reductase, depending upon the substrate. After a two hour reconstitution, the reductase eluted from hydroxylapatite at the location of holoreductase. It had little flavin dependence, was equimolar in FMN and FAD, and had nearly the specific activity (per mole flavin) of untreated reductase.^ The lack of activity and the ability of FMN to also reconstitute suggest that the redox center of FAD is essential for catalysis, rather than for structure. Dependence upon FAD is consistent with existing hypotheses for the catalytic cycle of the reductase. ^

Abundance and biomass of mesozooplankton from the Romanian littoral during DANUBS_2002 in spring and autumn 2002

The Danubs 2002 dataset contains zooplankton data collected in April, June,September and October 2002 in 11 station allong 5 transect in front of the Romanian littoral. Zooplankton sampling was undertaken at 11 stations where samples were collected using a Juday closing net in the 0-10, 10-25, and 25-50m layer (depending also on the water masses). The dataset includes samples analysed for mesozooplankton species composition and abundance. Sampling volume was estimated by multiplying the mouth area with the wire length. Taxon-specific mesozooplankton abundance was count under microscope. Total abundance is the sum of the counted individuals. Total biomass Fodder, Rotifera , Ctenophora and Noctiluca was estimated using a tabel with wet weight for each species an stage.

Ponte, Jorge Ricardo, La fragilidad de la memoria. Representaciones, prensa y poder de una ciudad latinoamericana en tiempos del modernismo. Mendoza, 1885/1910. Mendoza, Ed. Fundación CRICYT, 1999,450 p.

Fil: Arpini, Adriana.

Abundance of mesozooplankton in the SESAME Italian cruise S-IT2 in the Ionian Sea in March 2008

The "SESAME_IT2_ZooAbundance_0-50-100m_SZN" dataset contains data of mesozooplankton species composition and abundance (ind. m-3) from samples collected in the Ionian Sea in the late winter (2-8 March) of 2008 during the SESAME-WP2 cruise IT2. Samples were collected by vertical tows with a closing WP2 net (56 cm diameter, 200 ?m mesh size) in the following depth layers: 100-200 m, 50-100 m, 0-50 m. Sampling was always performed in light hours. A flowmeter was applied to the mouth of the net, however, due to its malfunctioning, the volume of filtered seawater was calculated by multiplying the the area by the height of the sampled layer from winch readings. After collection, each sample was split in two halves (1/2) after careful mixing with graduated beakers. Half sample was immediately fixed and preserved in a formaldehyde-seawater solution (4% final concentration) for species composition and abundance. The other half sample was kept fresh for biomass measurements (data already submitted to SESAME database in different files).Here, only the zooplankton abundance of samples in the upper layers 0-50 m and 50-100 m are presented. The abundance data of the samples in the layer 50-100 m will be submitted later in a separate file. The volume of filtered seawater was estimated by multiplying the the area by the height of the sampled layer from winch readings. Identification and counts of specimens were performed on aliquots (1/20-1/5) of the fixed sample or on the total sample (half of the original sample) by using a graduate large-bore pipette. Copepods were identified to the species level and separated into females, males and juveniles (copepodites). All other taxa were identified at the species level when possible, or at higher taxonomic levels. Taxonomic identification was done according to the most relevant and updated taxonomic literature. Total mesozooplankton abundance was computed as sum of all specific abundances determined as explained above.