Continuous-wave quantum yields of various cobalamins are influenced by competition between geminate recombination and cage escape

Quantum yields of photolysis of the cobalt-carbon bond for three cobalamin compounds were measured with a continuous-wave laser at 442 nm under both aerobic and anaerobic conditions. Aerobically, the initial homolysis product, Co(II) cobalamin, is trapped by oxygen to form aquocobalamin. Use of an excess of the radical trapping reagent 2,2,6,6-tetramethyl-1-piperidinyloxyl, under anaerobic conditions, scavenges the carbon radical and allows detection of the cobalt(II) photoproduct. Quantum yields measured under anaerobic conditions for 5'-deoxyadenosylcobalamin (phi (Co-C alpha),442 = 0.20 +/- 0.03) and methylcobalamin (phi (Co-C alpha),442 = 0.35 +/- 0.03) are in agreement with the values obtained under aerobic conditions (phi (Co-C alpha),442 = 0.19 +/- 0.04 and phi (Co-C alpha),442 = 0.36 +/- 0.04, respectively). Additionally, the quantum yield values for 5'-deoxyadenosylcobalamin and its base-off derivative (phi (Co-C alpha),442 = 0.045 +/- 0.015) match those obtained on a nanosecond time scale [Chen, E., & Chance, M. R. (1990) J. Biol. Chem. 256, 12987-12994]. A comparison of quantum yields obtained anaerobically for 5'-deoxyadenosylcobalamin and methylcobalamin in H2O versus ethylene glycol shows a 4-fold decrease for the former cobalamin and no change for the latter. These quantum yields are evaluated in terms of time-independent radical separation distances.     

Effects of the glucocorticoid agonist, RU28362, and the antagonist RU486 on lung phosphatidylcholine and antioxidant enzyme development in the genetically obese Zucker rat

The biochemical maturation of the lung in late gestation and in the young animal is regulated by glucocorticoids. The present study was aimed at dissociating the different glucocorticoid receptor sites involved in these regulatory functions. The obese Zucker rat was selected as a model for this study as it exhibits hypersensitivity to glucocorticoid hormone action by virtue of its elevated receptor numbers and activity. Two synthetic steroid analogues were administered to obese animals; RU28362, a specific type II receptor agonist, and the type II antagonist RU486. RU28362 promoted a strong catabolic effect, which was associated with reduced food intake and the abolition of growth in the rats. The agonist, RU28362, attenuated developmental increases in antioxidant enzyme activities, and altered the growth of the tissue. At the age studied, development of the lung phosphatidylcholine (PC) system was almost complete, but RU28362 increased disaturated PC 16:0/16:0 concentrations by almost 2-fold, and altered the molecular composition of total pulmonary PC. RU486 attenuated the growth of the rats and reduced their food intake. Treatment with the type II antagonist attenuated lung growth and increased the activities of pulmonary copper zinc (Cu/Zn) and manganese (Mn) superoxide dismutases. RU486 had no effect on lung PC concentrations and molecular composition. The data suggest a role for type I glucocorticoid receptors in the regulation of the antioxidant enzyme system in the lung, as type II antagonism will channel endogenous glucocorticoid binding to the type I site. Type II receptor binding would appear to play a role in regulating the lung PC content.     

Kinetic analysis of the coding properties of O6-methylguanine in DNA: the crucial role of the conformation of the phosphodiester bond

Production by N-nitroso compounds of O6-alkylguanine (O6-alkylG) in DNA directs the misincorporation of thymine during DNA replication, leading to G:C to A:T transition mutations, despite the fact that DNA containing O6-alkylG:T base pairs is less stable than that containing O6-alkylG:C pairs. We have examined the kinetics of incorporation by Klenow fragment (KF) of Escherichia coli DNA polymerase I of thymine (T) and of cytosine (C) opposite O6-MeG in the template DNA strand. Both T and C were incorporated opposite O6-MeG much slower than nucleotides forming regular A:T or G:C base pairs. Using various concentrations of dTTP, dCTP, or their phosphorothioate (Sp)-dNTP alpha S analogues, or a mixture of dTTP and dCTP, the progress of incorporation of a single nucleotide in a single catalytic cycle of a preformed KF-DNA complex was measured (pre-steady-state kinetics). The results were consistent with the kinetic scheme (Kuchta, R. D., Benkovic, P., & Benkovic, S. J. (1988) Biochemistry 27, 6716-6725): (1) binding of dNTP to polymerase-DNA; (2) conformational change in polymerase; (3) formation of phosphodiester between the dNTP and the 3'-OH of the primer; (4) conformational change of polymerase; (5) release of pyrophosphate. The results were analyzed mathematically to identify the steps at which the rate constants differ significantly between the incorporation of T and C. The only significant difference was the 5-fold difference in the rates of formation of the phosphodiester bond (for dTTP, kforward = 3.9 s-1 and kback = 1.9 s-1; for dCTP, kforward = 0.7 s-1 and kback = 0.9 s-1). These pre-steady-state progress curves were biphasic with a rapid initial burst followed by an apparently steady-state rise. Deconvolution of these curves gave direct evidence for the importance of the conformational change after polymerization by showing that the curves represented the sum of the rapid accumulation of the product of step 3 followed by the slow conversion of that to the product of step 5 (because of the rapidity of the release of pyrophosphate there was no significant accumulation of the product of step 4). The equilibrium constants for each step suggest that the greatest change in the Gibbs free energy occurs at the conformational change after polymerization and that while the formation of the phosphodiester bond to T is slightly exothermic, that to C is slightly endothermic.(ABSTRACT TRUNCATED AT 400 WORDS)     

Frictional assessment of magnesium stearate and palmitate lubricant powders

A method has been developed for the frictional assessment of powder lubricants using a shear-box tester similar to that employed by Jenike. One commercial magnesium stearate, three high-purity magnesium stearates and three high-purity magnesium palmitates were assessed using this method. Powders with a well-ordered crystal structure and particle shape have a lower initial maximum coefficient of friction μ_a. The difference between μ_a and μ_b, the equilibrium dynamic friction coefficient, gives an indication of lubricant film-forming propensity. The basic friction equation is obeyed by the powders for most of the load range studied but deviates slightly approaching zero normal load.     

Time-dependent optical spectroscopy and imaging for biomedicalapplications

The application of optical spectroscopy and imaging in living tissue is complicated by multiple scattering of light. In spectroscopy, this scattering causes uncertainty in the pathlength traveled by photons in the tissue, while images suffer reduced resolution and contrast. Picosecond light sources and fast detectors have made it possible to address these problems by direct measurement of the photon time-of-flight. Diffusion models of light propagation can be used to relate the measured distribution of photon transit times to the scattering and absorption coefficients of the tissue. The advantages of absolute absorption measurement are demonstrated for two problems: determination of hemoglobin oxygenation in tissue and in vivo measurement of the uptake of an exogenous chromosphere such as photosensitizer. Optical imaging may also be improved by the elimination of multiply scattered photons or by selective detection of photons arriving from a given region of the tissue. The potential advantages of these techniques are discussed and illustrated with experimental data     

Reaction of hexamethylphosphorous triamide with cotton cellulose

The reactivity of hexamethylphosphorus triamide, (P[N? (CH₃)₂]₃) (HPT), with cotton cellulose was examined under various conditions. HPT was found to react readily with the cellulose to produce a cellulose phosphite and/or phosphate ester having approximately one acidic hydrogen atom per phosphorus atom. The reaction of HPT with cotton cellulose was efficiently catalyzed by dimethylamine hydrochloride (1%). The phosphorus content imparted by the catalyzed reaction on cotton fabrics was approximately twice that imparted by the uncatalyzed reaction. Flame resistance of the treated fabrics ranged from fair to excellent. The treated fabrics also exhibited ion exchange properties and some crosslinking.     

Fluorescent heterogeneities in turbid media: limits for detection, characterization, and comparison with absorption

The fundamental limits for detection and characterization of fluorescent (phosphorescent) inhomogeneities embedded in tissuelike highly scattering turbid media are investigated. The absorption and fluorescence contrast introduced by exogenous fluorophores are also compared. Both analyses are based on practical signal-to-noise ratio considerations. For an object with fivefold fluorophore concentration and lifetime contrast with respect to the background tissue, we find the smallest detectable fluorescent object at 3-cm depth in tissuelike turbid media to be ~0.25 cm in radius, whereas the smallest characterizable object size is ~0.75 cm in radius, given a model with 1% amplitude and 0.5 degrees phase noise. We also find that, for fluorescence extinction coefficients epsilon 相似文献   

Fluorescent diffuse photon density waves in homogeneous and heterogeneous turbid media: analytic solutions and applications

We present analytic solutions for fluorescent diffuse photon density waves originating from fluorophores distributed in thick turbid media. Solutions are derived for a homogeneous turbid medium containing a uniform distribution of fluorophores and for a system that is homogeneous except for the presence of a single spherical inhomogeneity Generally the inhomogeneity has fluorophore concentration, and lifetime and optical properties that differ from those of the background. The analytic solutions are verified by numerical calculations and are used to determine the fluorophore lifetime and concentration changes required for the accurate detection of inhomogeneities in biologically relevant systems. The relative sensitivities of absorption and fluorescence methods are compared.     

Enhancing encoding capacity of combinational test stimulus decompressors

While scan-based compression is widely utilized in order to alleviate the test time and data volume problems,the overall compression level is dictated not only by the chain to channel ratio but also the ratio of encodable patterns.Aggressively increasing the number of scan chains in an effort to raise the compression levels may reduce the ratio of encodable patterns,degrading the overall compression level.In this paper,we present various methods to improve the ratio of encodable patterns.These methods are b...     

Rechtliche Fragen zur Praxis IT-forensischer Analysen in Organisationen

Der Einsatz von Informationstechnologie (IT) in Organisationen bietet viele M?glichkeiten des Missbrauchs. Besch?ftigte k?nnen sich die besonderen Eigenschaften der IT zunutze machen, um sich etwa in gesch?ftssch?digender Weise Vorteile zu verschaffen. Gleichzeitig erm?glicht IT aber auch, verbotene Handlungen aufzuspüren und zu belegen (IT-Forensik). Der Beitrag beschreibt die technische Praxis von IT-forensischen Analysen und geht auf damit verbundene rechtliche Fragestellungen ein.