Structure and function of type I copper in multicopper oxidases

The type I copper center in multicopper oxidases is constructed from 1Cys2His and weakly coordinating 1Met or the non-coordinating 1Phe/1Leu, and it exhibits spectral properties and an alkaline transition similar to those of the blue copper center in blue copper proteins. Since the type I copper center in multicopper oxidases is deeply buried inside the protein molecule, electron transfers to and from type I copper are performed through specific pathways: the hydrogen bond between an amino acid located at the substrate binding site and a His residue coordinating type I copper, and the His-Cys-His sequence connecting the type I copper center and the trinuclear copper center comprised of a type II copper and a pair of type III coppers. The intramolecular electron transfer rates can be tuned by mutating the fourth ligand of type I copper. Further, mutation at the Cys ligand gives a vacant type I copper center and traps the reaction intermediate during the four-electron reduction of dioxygen.     

Decreased activities of liver microsomal drug-metabolizing enzymes in the rats bearing walker carcinosarcoma

Riassunto Ratti portatori di carcinosarcoma di Walker, presentano una notevole diminuzione di attività di quel gruppo di enzimi microsomici epatici da cui dipende il metabolismo di una numerosa serie di farmaci. I nostri rilievi sono stati fatti prendendo in considerazione il metabolismo dell'esobarbital, della stricnina e del meprobamato.     

Carbon tetrachloride modulates the rat hepatic microsomal UDP-glucuronyl transferase activity and membrane fluidity

Summary Modulations in rat hepatic microsomal UDP-glucuronyl transferase activity have been observed during carbon tetrachloride (CCl₄) poisoning, with a large decrease in the enzyme cooperativity and increase in the membrane fluidity, occurring 30 min after administration. The results strengthen the possibility that an increase in microsomal membrane fluidity may be an early key event in liver injury induced by CCl₄.Acknowledgments. This work was supported by funds of the University of Athens.     

Interaction of organic dyes with hepatic microsomal drug-metabolizing monoöxygenases in vitro

Summary Organic dyes such as malachite green, methylene blue, fuchsin, safranine T, neutral red, phenazine methosulphate, riboflavin, dichlorophenolindophenol, phenolphthalein, and fluorescein, inhibit hepatic microsomal mixedfunction oxidases and, partly, enhance, partly, inhibit hepatic microsomal NADPH-dependent cytochrome c and neotetrazolium reductases, in contrast to other inhibitors of drug metabolism which do not affect cytochrome c reductase but only interact with cytochrome P-450.Dedicated to Prof. G. Pfleiderer on the occasion of his 60th birthday.Acknowledgment. The author thanks Mrs Fey, Mrs Meier, and Mr Weinrauch, for their skilful technical assistance; he is grateful to Dr A. Sinharay (Hoechst AG) for providing him with anisic ester and methylayapanine.     

Inhibition of cytosolic rat hepatic glutathione S-transferase activities by bromosulfophthaleins

Summary The enzymatically catalyzed conjugation of glutathione with 1-chloro-2,4-dinitrobenzene and styrene-7,8-oxide is inhibited by tetrabromosulfophthalein in a non-competetive way, although tetrabromosulfophthalein itself is a substrate for glutathione S-transferases. Dibromosulfophthalein, which is not a substrate for glutathione S-transferases, inhibits the 1-chloro-2,4-dinitrobenzene conjugation competetively and the styrene-7,8-oxide conjugation non-competetively.     

Diphasic action of the aliphatic amide,HOE 17879, on hepatic microsomal drug metabolizing enzymes in the mouse

Summary -Hydroxy--ethylbutyryl-diethylamide (HOE 17879) influences the hepatic microsomal drug hydroxylating enzyme system of mice in a two-phasic way: First it reduces the overall reaction of drug hydroxylation but not the single enzymes whilst in the second period it increases enzyme activities.Acknowledgments. The authors are indebted to their colleague, Dr.K. Schmitt, for making the substance HOE 17879 available. They wish to thank to Mr.Mayer, Mr.Paulusch Mrs.Fey, Mrs.Stein, and MissFehlings for their skilful assistance in the project.     

Release of oxidases from the roots of plants

Summary In this paper, we report on the isolation and purification of the oxidase released from the roots ofLactuca sativa seedlings. This is the first time that the release of oxidases from the roots of plants into their rhizosphere has been reported.     

Electron microscopic studies on hepatic cells in mice after the administration of glutathione mixtures

Zusammenfassung Anschwellung und Lysis von Mitochondrien im Reagenzglas (in vivo), durch Zugabe von Glutathiongemisch zur Nährlösung verursacht, wurde elektronenmikroskopisch geprüft. Die Mitochondrien wurden in Mäuseleberzellen nach i.p. Injektion zusammen mit dem Glutathiongemisch untersucht.     

The absence of melanopsin alters retinal clock function and dopamine regulation by light

The retinal circadian clock is crucial for optimal regulation of retinal physiology and function, yet its cellular location in mammals is still controversial. We used laser microdissection to investigate the circadian profiles and phase relations of clock gene expression and Period gene induction by light in the isolated outer (rods/cones) and inner (inner nuclear and ganglion cell layers) regions in wild-type and melanopsin-knockout (Opn ₄ ^?/? ) mouse retinas. In the wild-type mouse, all clock genes are rhythmically expressed in the photoreceptor layer but not in the inner retina. For clock genes that are rhythmic in both retinal compartments, the circadian profiles are out of phase. These results are consistent with the view that photoreceptors are a potential site of circadian rhythm generation. In mice lacking melanopsin, we found an unexpected loss of clock gene rhythms and of the photic induction of Per1-Per2 mRNAs only in the outer retina. Since melanopsin ganglion cells are known to provide a feed-back signalling pathway for photic information to dopaminergic cells, we further examined dopamine (DA) synthesis in Opn ₄ ^?/? mice. The lack of melanopsin prevented the light-dependent increase of tyrosine hydroxylase (TH) mRNA and of DA and, in constant darkness, led to comparatively high levels of both components. These results suggest that melanopsin is required for molecular clock function and DA regulation in the retina, and that Period gene induction by light is mediated by a melanopsin-dependent, DA-driven signal acting on retinal photoreceptors.     

Nicotinamide desamidase in the culture medium of neuroblastoma cells]

Important amounts of nicotinic acid appear in the growth medium of neuroblastoma cell cultures. It is shown that an enzyme released by the cells into the growth medium deamidates the nicotinamide supplied by the growth medium to nicotinic acid.     

Changes in efferent activities of the gastric vagus nerve by administration of glucose in the portal vein

Summary Injection of glucose in the portal vein, as well as in the left carotid artery, brought a transient decrease in efferent discharges of the gastric vagus nerve, whereas venous injection of regular insulin brought a gradual increase in the discharges.Acknowledgment. The authors thank Prof. A. Niijima for his helpful advice.     

Inhibition of thyroid function following the administration of metopiron (SU-4885)

Zusammenfassung Die Goitrogen-Wirkung des Metopirons (SU-4885, CIBA) wurde in Feld- und Wüstenmaus mittels radioaktiven I¹³¹ untersucht. Das radioiodingebundene Protein (Pb I¹³¹) war nach SU-4885 vermindert. Die Zunahme der gesamten Radioaktivität in der Schilddrüse lag bei 49%. Metopiron wirkt als Schilddrüsen-Hemmer.

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We are grateful to Prof.P. N. Srivastava for providing facilities. The investigation was supported by University Grants Commission, New Delhi, India.     

Targeting NADPH oxidases for the treatment of cancer and inflammation

NADPH oxidases are a family of oxidases that utilize molecular oxygen to generate hydrogen peroxide and superoxide, thus indicating physiological functions of these highly reactive and short-lived species. The regulation of these NADPH oxidases (nox) enzymes is complex, with many members of this family exhibiting complexity in terms of subunit composition, cellular location, and tissue-specific expression. While the complexity of the nox family (Nox1-5, Duox1, 2) is daunting, the complexity also allows for targeting of NADPH oxidases in disease states. In this review, we discuss which inflammatory and malignant disorders can be targeted by nox inhibitors, as well as clinical experience in the use of such inhibitors.     

Changes in efferent activities of the gastric vagus nerve by administration of glucose in the portal vein.

Injection of glucose in the portal vein, as well as in the left carotid artery, brought a transient decrease in efferent discharges of the gastric vagus nerve, whereas venous injection of regular insulin brought a gradual increase in the discharges.     

Inhibitory effects of nitrososarcosine on mouse liver mixed function oxidase activity

Zusammenfassung Nachweis, dass Nitrosarcosin im Gegensatz zu Dimethyl- und Dibutylnitrosamin die nichtfunktionelle Oxidase in der Mäuseleber hemmt.

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This work was supported by N.I.E.H.S. grant No. ES 00713.

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We are indebted to the technical help of Ms.Heather McClanahan.