Ferredoxin reductase catalyzes styrene oxidation to styrene oxide

Summary The flavoprotein ferredoxin reductase catalyzed the oxidation of styrene to styrene oxide in the presence of NADPH. This reaction was inhibited by the addition of catalase and superoxide dismutase. The addition of the nonheme iron protein ferredoxin partially inhibited styrene oxidation. H₂O₂ was also able to catalyze this reaction when added to the enzyme in the absence of NADPH.Acknowledgments. This work was supported by C.N.R. (National Research Council), Rome, Italy contract No. 79.03197.04.     

Elektronenspinresonanzmessungen zur Untersuchung von Kinetik und Mechanismus von Cu2+-katalysierten Reaktionen

Summary Cu²⁺-complexes with different monodentate ligands PYR, e.g. pyridine, 2,4,6-collidine and imidazole, catalyse the oxidation ofo-phenylenediamine (H₂B) to 3,5-dihydro-2-amino-3-iminophenazine (PHEN) by O₂. Investigation of the electron paramagnetic resonance during reaction gives interesting details on the function of Cu²⁺ as a catalyser. The formation of mixed complexes (H₂B)Cu²⁺(PYR) and its influence on the reaction rated[PHEN]/dt is demonstrated. In the ratedetermining reaction, Cu²⁺ is reduced to Cu⁺, which is reoxidized by O₂. During reaction the ratio [Cu²⁺]/[Cu⁺] is determined by means of e.p.r. measurements.     

Die katalytische Mutation der Ionen Al3+ und MoO

Summary The catalytic mutation of the ions Al³⁺ and MoO ₄ ^2– on the mixed carrier Cd(OH)₂/Co(OH)₂(Tr) is recognizable by the fact that the combination Tr+Al³⁺+MoO ₄ ^2– is more active in the decomposition of H₂O₂ than the combination Tr+MoO ₄ ^2– +Al³⁺.     

Hydrogen peroxide generation inTrypanosoma cruzi

Summary Homogenates from T. cruzi epimastigotes produced 3.4 pmoles H₂O₂/min 10⁶ cells, as detected by the cytochrome c peroxidase assay. Addition of NADH or NADPH increased H₂O₂ production by a factor of 3 and 5, respectively. When supplemented with NADH and NADPH, the mitochondrial, microsomal, and supernatant fractions produced H₂O₂, the soluble fraction and the mitochondrial membranes being apparently the main generators of H₂O₂. The epimastigote homogenates showed cyanide-sensitive superoxide dismutase activity, equivalent to 0.28 g bovine superoxide dismutase per mg homogenate protein.This investigation was supported by grants from Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET) Argentina and the Scientific Office, American States Organization.Career Investigator of CONICET.     

Non-linear kinetics of microsomal styrene monooxygenase after phenobarbital pre-treatment

Summary Pretreatment of rats with phenobarbital, but not with 3-methylcholanthrene, induces liver microsomal styrene monooxygenase. Under these conditions the kinetic profile is not linear and can be divided into 2 distinct curves. Evidence is presented indicating that the combined treatment with phenobarbital and CoCl₂ destroy the high affinity enzyme, suggesting that the native cytochrome is less sensitive to the action of CoCl₂.This paper was supported by grant No. 181-77-ENVI I awarded by EEC.     

Über Struktur und Aktivität der den H2O2-Zerfall katalysierenden Cu2+-Komplexe. — VI. Differenzierung von nativer RNS und nativer DNS bzw. denaturierter DNS auf Grund der katalytischen Eigenschaften

Summary The catalysis of H₂O₂ decomposition by Cu²⁺-complexes of RNA and DNA has been investigated. It is shown that both complexes decompose H₂O₂, but only the Cu²⁺-RNA-system shows peroxidative activity too, e.g. only in this case the nucleotide bases are degraded. Thermal denaturation of DNA also leads to a Cu²⁺-complex with peroxidative activity, the latter being dependent on the degree of denaturation.

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V. Mitteilung:S. Petri, H. Sigel undH. Erlenmeyer, Helv. chim. Acta49, 1778 (1966).

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12. Mitteilung überMetallionen und H ₂ O ₂; 11. Mitteilung:H. Ch. Curtius, P. Anders, R. Zell, H. Sigel undH. Erlenmeyer, Helv. chim. Acta49, 2256 (1966).     

Radical yields in X-irradiated aqueous solutions

Résumé La valeur deGh+Goh peut être obtenue en utilisant des solutions aqueuses de certains composés organiques contenant Cu²⁺ et/ou Fe³⁺. Les ions métalliques sont réduits par H et par les radicaux produits par l'oxydation des composés organiques par OH. A concentration convenable des réactifs, on a;G _Fe ²⁺ =G _H+G _OH.     

Comparative study of oxalate oxidase in three genotypes ofSorghum vulgare

Summary The presence of an oxalate oxidase (EC 1.2.3.4) has been demonstrated in 15,000×g supernatants prepared from 10-day-old seedlings of three genotypes ofSorghum vulgare: grain sorghum hybrid (CSH-5), grain-cum-forage sorghum (PC-6) and forage sorghum (PC-1). The specific activity of the enzyme in the different tissues of seedlings was found to be present in the order leaves > stems > roots in PC-6 and PC-1, but this order was reversed in CSH-5. A comparison of the different properties of the leaf enzyme of these three genotypes of sorghum revealed that the enzyme has maximum activity in the acidic pH range from 4.0 to 5.0 and in the temperature range from 37°C to 40°C. The enzyme was stimulated by Cu²⁺ and Fe²⁺. The rate of H₂O₂ formation in the enzyme reaction was linear up to 5 min and was stoichiometrically related to oxalate consumption. The enzyme is unaffected by Na⁺ at physiological concentration (0.15 M). The superiority of this enzyme over moss and other plant enzymes for enzymic determination of urinary oxalate is discussed.     

Effects of scavengers of superoxide radicals,hydrogen peroxide,singlet oxygen and hydroxyl radicals on malondialdehyde generation from arachidonic acid by bovine seminal vesicle microsomes

Summary Superoxide dismutase, catalase and sodium formate did not inhibit the formation of malondialdehyde (MDA) from arachidonic acid, suggesting that O ₂ , H₂O₂ and OH^. are not involved in the enzymatical oxidation of arachidonic acid. Sodium azide was found to be an inhibitor of MDA production.     

Kinetic behaviour of microsomal styrene monooxygenase and styrene epoxide hydratase in different animal species

Summary The apparent K_m and V_max values of styrene epoxide forming monooxygenase and styrene epoxide hydratase have been evaluated in the liver microsomes of male rats, mice, guinea-pigs and rabbits. Epoxide hydratase gave much higher and more uniform K_m values than the monooxygenase in the species considered.Acknowledgments. This work has been partially supported by the Fondazione Valenti (Milano). To whom enquiries should be addressed.     

Induction of nuclear styrene monooxygenase and epoxide hydrolase in rat liver

Summary The apparent K_m and V_max of styrene monooxygenase and styrene epoxide hydrolase were determined in intact nuclear preparations from male rat liver after in vivo treatment with phenobarbital and -naphthoflavone, which are known to induce microsomal cytochrome P-450 and cytochrome P-448 respectively. Treatment with phenobarbital does not alter the apparent K_m, but greatly increases the V_max of both nuclear styrene monooxygenase and styrene epoxide hydrolase. Almost the same pattern is observed for styrene monooxygenase after treatment with -naphthoflavone, whereas the same treatment slightly increases both the V_max and K_m value of styrene epoxide hydrolase.This work was supported by the CNR (National Research Council) Contract No. 78.02864.96 within the special program Control of Cancer Growth.     

Functional and biochemical characteristics of mitochondrial fractions from rat liver in cold-induced oxidative stress

We determined characteristics of rat liver mitochondrial fractions, resolved at 1000 (M₁), 3000 (M₃), and 10,000 g (M₁₀) after 2 and 10 days cold exposure. In all groups, the M₁ fraction exhibited the highest oxidative capacity, oxidative damage, H₂O₂ production rate, and susceptibility to stress conditions, and the lowest antioxidant levels. Cold exposure increased cytochrome oxidase activity in all fractions and succinate-supported O₂ consumption in the M₁ and M₁₀ fractions during state 3 and state 4 respiration, respectively. With succinate, the H₂O₂ release rate increased in all fractions during state 4 and state 3 respiration, whereas with pyruvate/malate, it increased only during state 4 respiration. Increases in tissue mitochondrial proteins caused a faster H₂O₂ flow from the mitochondrial to cytosolic compartment, which was limited by the reduction in the M₁ fraction. Despite increased liposoluble antioxidant levels, cold also caused enhanced oxidative damage and susceptibility to oxidative challenge and Ca²⁺-induced swelling in all fractions. These changes leading to elimination of H₂O₂-overproducing mitochondria avoided excessive tissue damage. We propose that triiodothyronine, whose levels increase in the cold environment, brings about the biochemical changes producing oxidative damage and those limiting its extent.Received 16 July 2004; received after revision 27 September 2004; accepted 18 October 2004     

Strukturspezifischer Abbau von Polypeptid-Metall-Komplexen. V. Abbau des Cu2+-Polymyxin-B-Komplexes durch NH2OH

Summary NH₂OH as well as H₂O₂ (previous results^3,4) decompose the Cu₂₊ complex of polymyxin B as can be shown with thin layer chromatography; the spots of the peptide fragments of both reactions were compared.     

Involvement of reactive oxygen species in the microsomal S-oxidation of thiobenzamide

Summary Superoxide dismutase, catalase and methional proved capable of inhibiting the microsomal oxidation of thiobenzamide, which is most probably catalyzed by the flavin-containing monooxygenase. This indicates that excited oxygen species (e. g.·O ₂ ^– , H₂O₂, ·OH) are involved in the catalytic cycle of this enzymatic reaction. CO, which inhibits the cytochrome P-450-dependent oxygen radical formation, had no effect on the oxidation reaction, suggesting that the source of the reactive oxygen species is not the microsomal mixed-function oxidase.     

Obtention d'un dérivé du sclaréol à partir de l'ambréine

Summary One of the products of the chromic acid oxidation of the animal triterpene ambreine (V) has been identified with the lactone C₁₆H₂₆O₂ (III) whichRuzicka and coll. had isolated by oxidation of the plant diterpene sclareol (II). This identification confirms recent findings of the authors and ofRuzicka, Dürst andJeger and shows that the hydroxyle of ambreine (V) is at the same place as the hydroxyle of sclareol which is attached to the perhydronaphtalene ring.

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4^me communication sur les constituants de l'ambre gris     

Biochemical events associated with rapid cellular damage during the oxygen-and calcium-paradoxes of the mammalian heart

Summary The O_2– and Ca²⁺-paradoxes have a number of features in common and it is suggested that release of cytosolic proteins in both paradoxes is initiated by the activation of a sarcolemma NAD(P)H dehydrogenase which can generate a transmembrane flow of H⁺ and e^– and also oxygen radicals or recox cycling which damage ion channels and membrane proteins (phase I). Entry of Ca²⁺ through the damaged ion channels then exacerbates the damage by further activating this system, either directly or indirectly, and the redox cycling and/or oxygen radicals cause further damage to integral and cytoskeletal proteins of the sarcolemma resulting in microdamage to the integrity of the membrane (phase II) and the consequent release or exocytosis of cytoplasmic proteins and, under specialised condition, the blebbing of the sarcolemma. The system may be primed either by removal of extracellular Ca²⁺ or by raising [Ca²⁺]_i by a variety of measures, these two actions being synergistic. The system is initially activated in the Ca²⁺-paradox by the membrane perturbation associated with removal of extracellular Ca²⁺; prolonged anoxia in the metabolically active cardiac muscle causes a depletion of the ATP supply, particularly in the absence of glucose, and hence a rise in [Ca²⁺]_i in phase I of the oxygen paradox with the consequent activation of the NAD(P)H oxidase at the sarcolemma. Oxygen radicals are probably generated in both paradoxes and may have a partial role in the genesis of damage, but are not essential in the Ca²⁺-paradox which continues under anoxia. Massive entry of Ca²⁺ also activates an intracellularly localised dehydrogenase (probably at the SR) which produces myofilament damage by redox cycling.     

Bactericidal activity againstPseudomonas aeruginosa is acquired by cultured human monocyte-derived macrophages after uptake of myeloperoxidase

Myeloperoxidase (MPO) is an enzyme located within polymorphonuclear neutrophils capable of producting cytotoxic oxidant species that are particularly active against bacteria with polysaccharide capsules.Pseudomonas aeruginosa (10⁶ bacteria per 1 ml) are killed within 1 h in vitro by a MPO/H₂O₂/Cl^– system (48 mU=132 ng of MPO). The question arose as to whether human macrophages would acquire cytotoxic activity when loaded with this enzyme. Monocytes were therefore isolated from human blood and cultured for up to ten days to induce maturation to macrophages. These cells lost endogenous MPO within five days while H₂O₂ production in response to stimulation by phorbol myristate acetate (10^–6M) decreased to 23% within ten days. On the other hand, their capacity to take up exogenous MPO increased fourfold from day three to day ten. Human macrophages cultured from eight days (when both H₂O₂ production and MPO uptake were sufficient) were therefore used to study the effects of MPO uptake on cytocidal activity againstPseudomonas aeruginosa. After a 1 h MPO loading period, macrophages (5×10⁵ cells per ml) were incubated in the presence of bacteria (0.5 to 2×10⁶ bacteria per ml) for 2 h at 37°C. At a bacteria/macrophage ratio of 1, only 34.8±7.0% of bacteria survived (compared to killing by non-loaded macrophages), while 74.4±9.3% survived at a ratio of 4. From these results, we conclude that loading macrophages with exogenous MPO could enhance their microbicidal activity, suggesting a potentially useful therapeutic application.     

Alcaloïde des Canneberges du Nouveau-Brunswick

Summary A new alkaloid C₂₀H₂₂O₂N₂ (cannagunine) obtained from canadian Vaccinium Oxycoccus is shown to have structure 1. The structure of this compound was supported by NMR-, SM-, IR- and UV-spectra.     

Ginkgo biloba extract inhibits oxygen species production generated by phorbol myristate acetate stimulated human leukocytes

Summary A Ginkgo biloba extract (Gbe) containing flavonoids, among other compounds, was tested for the release of activated oxygen species ( , H₂O₂, OH^.) during the stimulation of human neutrophils (PMNs) by a soluble agonist. The extract slows down O₂-consumption (respiratory burst) of stimulated cells by its inhibitory action on NADPH-oxidase, the enzyme responsible for the reduction of O₂ to . Consequently, superoxide anion ( ) and hydrogen peroxide (H₂O₂) production is significantly decreased when the PMNs stimulation is done in the presence of the extract at concentrations of 500, 250 and 125 g/ml. Moreover, the hydroxyl radical generation (OH^.) is very much decreased at concentrations as low as 15.6 g Gbe/ml, which indicates that the extract also has free radical scavenging activity. Gbe is able at least to reduce very severel the activity of myeloperoxidase contained in neutrophils. This enzyme, secreted into the intra and extracellular medium, catalyzes the oxidation of chloride (Cl^–) by H₂O₂ to yield strong oxidants (HOCl, chloramines) which are implicated in inflammatory processes