Kinetics of Reduction of Cytochrome \u3ci\u3ec\u3c/i\u3e Oxidase by Dithionite and the Effect of Hydrogen Peroxide

The reduction of cytochrome c oxidase by dithionite was reinvestigated with a flow-flash technique and with varied enzyme preparations. Since cytochrome a3 may be defined as the heme in oxidase which can form a photolabile CO adduct in the reduced state, it is possible to follow the time course of cytochrome a3 reduction by monitoring the onset of photosensitivity. The onset of photosensitivity and the overall rate of heme reduction were compared for Yonetani and Hartzell-Beinert preparations of cytochrome c oxidase and for the enzyme isolated from blue marlin and hammer-head shark. For all of these preparations the faster phase of heme reduction, which is dithionite concentration-dependent, is almost completed when the fraction of photosensitive material is still small. We conclude that cytochrome a3 in the resting enzyme is consistently reduced by an intramolecular electron transfer mechanism. To determine if this is true also for the pulsed enzyme, we examined the time course of dithionite reduction of the peroxide complex of the pulsed enzyme. It has been previously shown that pulsed cytochrome c oxidase can interact with H2O2 and form a stable room temperature peroxide adduct (Bickar, D., Bonaventura, J., and Bonaventura, C. (1982) Biochemistry 21, 2661-2666). Rather complex kinetics of heme reduction are observed when dithionite is added to enzyme preparations that contain H2O2. The time courses observed provide unequivocal evidence that H2O2 can, under these conditions, be used by cytochrome c oxidase as an electron acceptor. Experiments carried out in the presence of CO show that a direct dithionite reduction of cytochrome a3 in the peroxide complex of the pulsed enzyme does not occur

Cannabinoids in the treatment of epilepsy: current status and future prospects

Cannabidiol (CBD) is one of the prominent phytocannabinoids found in Cannabis sativa, differentiating from Δ9-tetrahydrocannabinol (THC) for its non-intoxicating profile and its antianxiety/antipsychotic effects. CBD is a multi-target drug whose anti-convulsant properties are supposed to be independent of endocannabinoid receptor CB1 and might be related to several underlying mechanisms, such as antagonism on the orphan GPR55 receptor, regulation of adenosine tone, activation of 5HT1A receptors and modulation of calcium intracellular levels. CBD is a lipophilic compound with low oral bioavailability (6%) due to poor intestinal absorption and high first-pass metabolism. Its exposure parameters are greatly influenced by feeding status (ie, high fatcontaining meals). It is mainly metabolized by cytochrome P 450 (CYP) 3A4 and 2C19, which it strongly inhibits. A proprietary formulation of highly purified, plant-derived CBD has been recently licensed as an adjunctive treatment for Dravet syndrome (DS) and Lennox-Gastaut syndrome (LGS), while it is being currently investigated in tuberous sclerosis complex. The regulatory agencies’ approval was granted based on four pivotal double-blind, placebocontrolled, randomized clinical trials (RCTs) on overall 154 DS patients and 396 LGS ones, receiving CBD 10 or 20 mg/kg/day BID as active treatment. The primary endpoint (reduction in monthly seizure frequency) was met by both CBD doses. Most patients reported adverse events (AEs), generally from mild to moderate and transient, which mainly consisted of somnolence, sedation, decreased appetite, diarrhea and elevation in aminotransferase levels, the last being documented only in subjects on concomitant valproate therapy. The interaction between CBD and clobazam, likely due to CYP2C19 inhibition, might contribute to some AEs, especially somnolence, but also to CBD clinical effectiveness. Cannabidivarin (CBDV), the propyl analogue of CBD, showed anti-convulsant properties in pre-clinical studies, but a plant-derived, purified proprietary formulation of CBDV recently failed the Phase II RCT in patients with uncontrolled focal seizures

Dynamic models for Large Eddy Simulation of compressible flows with a high order DG method

The impact of dynamic models for applications to LES of compressible flows is assessed in the framework of a numerical model based on high order discontinuous finite elements. The projections onto lower dimensional subspaces associated with lower degree basis functions are used as LES filter, along the lines proposed in Variational Multiscale templates. Comparisons with DNS results available in the literature for plane and constricted channel flows at Mach numbers 0.2, 0.7 and 1.5 show clearly that the dynamic models are able to improve the prediction of most key features of the flow with respect to the Smagorinsky models employed so far in a VMS-DG context

Keck Imaging of Binary L Dwarfs

We present Keck near-infrared imaging of three binary L dwarf systems, all of which are likely to be sub-stellar. Two are lithium dwarfs, and a third exhibits an L7 spectral type, making it the coolest binary known to date. All have component flux ratios near 1 and projected physical separations between 5 and 10 AU, assuming distances of 18 to 26 pc from recent measurements of trigonometric parallax. These surprisingly similar binaries represent the sole detections of companions in ten L dwarf systems which were analyzed in the preliminary phase of a much larger dual-epoch imaging survey. The detection rate prompts us to speculate that binary companions to L dwarfs are common, that similar-mass systems predominate, and that their distribution peaks at radial distances in accord both with M dwarf binaries and with the radial location of Jovian planets in our own solar system. To fully establish these conjectures against doubts raised by biases inherent in this small preliminary survey, however, will require quantitative analysis of a larger volume-limited sample which has been observed with high resolution and dynamic range.Comment: LaTex manuscript in 13 pages, 3 postscript figures, Accepted for publication in the Letters of the Astrophysical Journal; Postscript pre-print version available at: http://www.hep.upenn.edu/PORG/papers/koerner99a.p

Effect of heme and non-heme ligands on subunit dissociation of normal and carboxypeptidase-digested hemoglobin. Gel filtration and flash photolysis studies.

The dissociation of normal and carboxypeptidase-digested human hemoglobin has been studied by gel filtration under several experimental conditions. These include (a) different derivatives, notably deoxy-, oxy-, and CO-hemoglobin, (b) changes in solvent composition and in pH, and (c) addition of inositol hexaphosphate. In normal hemoglobin, in agreement with previous results, the deoxygenated derivative is much less dissociated than the oxy or CO ones. This difference is observed also in some of the digested hemoglobins, but tends to vanish in those proteins in which, as a result of extensive digestion, the conformational change accompanying ligand binding is abolished. The dissociation of normal and digested hemoglobins is affected by solvent composition, is at a minimum at pH near 8, and is decreased by the addition of inositol hexaphosphate. Parallel flash photolysis experiments, performed under conditions identical with those used in the gel filtration studies, indicate that the appearance of quickly reacting material parallels dissociation into dimers in normal hemoglobin. Both in normal and digested hemoglobins conditions which decrease dissociation decrease the fraction of rapidly reacting material. In the digested hemoglobins the fraction of rapidly reacting material may be much higher than can be accounted for by the amount of dimers, indicating in these cases that the tetramers may be rapidly reacting. The data point once again to the critical role of the COOH-terminal residues in maintaining the subunit structure of hemoglobin and the interaction effects associated with it

The Forecasting Performance of Dynamic Factor Models with Vintage Data

We present a comparative analysis of the forecasting performance of two dynamic factor models, the Stock and Watson (2002a, b) model and the Forni, Hallin, Lippi and Reichlin (2005) model, based on vintage data. Our dataset that contains 107 monthly US “first release” macroeconomic and financial vintage time series, spanning the 1996:12 to 2017:6 period with monthly periodicity, extracted from the Bloomberg database† . We compute real-time one-month-ahead forecasts with both models for four key macroeconomic variables: the month-on-month change in industrial production, the unemployment rate, the core consumer price index and the ISM Purchasing Managers’ Index. First, we find that both the Stock and Watson and the Forni, Hallin, Lippi and Reichlin models outperform simple autoregressions for industrial production, unemployment rate and consumer prices, but that only the first model does so for the PMI. Second, we find that neither models always outperform the other. While Forni, Hallin, Lippi and Reichlin’s beats Stock and Watson’s in forecasting industrial production and consumer prices, the opposite happens for the unemployment rate and the PMI