Rapid characterization and partial purification of various animal amine oxidases

Summary The use of chromatofocusing to obtain a rapid characterization of tissue amine oxidases from various mammals is proposed. This technique yields partially purified enzymes well suited for immunological studies. Chromatofocusing can be also used in a three-step purification of pig kidney diamine oxidase.Acknowledgments. This work has been in part supported by CNR, Special Project Chimica Fine Contract No. 80.02.107.95.     

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     

Synthesis of nuclear and mitochondrial DNA in rat liver after injection of the carcinogenic compound diethylnitrosamine

Résumé L'injection de diéthylnitrosamine à des rats provoque, dans les heures qui suivent, une inhibition de synthèse plus forte au niveau du DNA nucléaire du foie qu'au niveau du DNA mitochondrial. A la phase d'inhibition succède une stimulation de synthèse de nDNA, avec retour à un niveau normal de la synthèse de mtDNA.

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Grants from the Fonds de la Recherche Scientifique Médicale and Fonds de la Recherche Fondamentale Collective are gratefully acknowledged. The authors thank MissR. Lemestrée for skillfull technical assistance.     

Effect of some monoamine oxidase inhibitors on deamination of biogenic monoamines by rat liver mitochondrial monoamine oxidase

N--N- MAO , , .     

Separation of germ layers in presomite rat embryos

Résumé Les feuillets du cylindre-Æuf du rat ont été disjoints par une solution composée de deux enzymes: 0,5% de trypsine et 2,5% de pancréatine. La séparation a été achevée par une aiguille de tungstène. Les feuillets ainsi détachés retiennent leur pouvoir de croissance et de différenciation comme homogreffes, sous la capsule rénale.     

On the radio-sensitivity of rat liver

Résumé Les lobes antérieurs du foie de rats ont été prélevés et exposés aux rayons-X jusqu'à des doses de 91 kR et en employant comme indicateur biochimique de survivance l'incorporation de³H-leucine. Une dose de 45 kR fit diminuer l'incorporation au cours des 3 premiers jours suivant l'exposition; ce type de blessure de radiation nécessite donc plusieurs jours pour se manifester. Ces rêsultats montrent qu'au commencement de l'interphase l'hépatocyte est extrêmement résistant à la mort par radiation.     

Antigen-distribution in rat liver mitochondria

Riassunto I mitocondri di fegato di ratto possiedono almeno tre gruppi di antigeni, dei quali uno è esclusivamente legato alle membrane mitocondriali, uno è presente nella frazione solubile in 0,3% desossicolato di sodio ed un altro, che, sebbene principalmente localizzato nelle membrane, è anche dimostrabile nella frazione desossicolato solubile.     

Regulation of mitochondrial aconitase by phosphorylation in diabetic rat heart

Mitochondrial dysfunction and protein kinase C (PKC) activation are consistently found in diabetic cardiomyopathy but their relationship remains unclear. This study identified mitochondrial aconitase as a downstream target of PKC activation using immunoblotting and mass spectrometry, and then characterized phosphorylation-induced changes in its activity in hearts from type 1 diabetic rats. PKCβ₂ co-immunoprecipitated with phosphorylated aconitase from mitochondria isolated from diabetic hearts. Augmented phosphorylation of mitochondrial aconitase in diabetic hearts was found to be associated with an increase in its reverse activity (isocitrate to aconitate), while the rate of the forward activity was unchanged. Similar results were obtained on phosphorylation of mitochondrial aconitase by PKCβ₂ in vitro. These results demonstrate the regulation of mitochondrial aconitase activity by PKC-dependent phosphorylation. This may influence the activity of the tricarboxylic acid cycle, and contribute to impaired mitochondrial function and energy metabolism in diabetic hearts. Received 31 October 2008; received after revision 17 December 2008; accepted 2 January 2009