Effects of myocardial ischemia and reperfusion on mitochondrial function and susceptibility to oxidative stress

We investigated the effects of ischemia duration on the functional response of mitochondria to reperfusion and its relationship with changes in mitochondrial susceptibility to oxidative stress. Mitochondria were isolated from hearts perfused by the Langendorff technique immediately after different periods of global ischemia or reperfusion following such ischemia periods. Rates of O₂ consumption and H₂O₂ release with complex I- and complex II-linked substrates, lipid peroxidation, overall antioxidant capacity, capacity to remove H₂O₂, and susceptibility to oxidative stress were determined. The effects of ischemia on some parameters were time dependent so that the changes were greater after 45 than after 20 min of ischemia, or were significantly different to the nonischemic control only after 45 min of ischemia. Thus, succinate-supported state 3 respiration exhibited a significant decrease after 20 min of ischemia and a greater decrease after 45 min, while pyruvate malate-supported respiration showed a significant decrease only after 45 min of ischemia, indicating an ischemia-induced early inhibition of complex II and a late inhibition of complex I. Furthermore, both succinate and pyruvate malate-supported H₂O₂ release showed significant increases only after 45 min of ischemia. Similarly, whole antioxidant capacity significantly increased and susceptibility to oxidants significantly decreased after 45 min of ischemia. Such changes were likely due to the accumulation of reducing equivalents, which are able to remove peroxides and maintain thiols in a reduced state. This condition, which protects mitochondria against oxidants, increases mitochondrial production of oxyradicals and oxidative damage during reperfusion. This could explain the smaller functional recovery of the tissue and the further decline of the mitochondrial function after reperfusion following the longer period of oxygen deprivation. Received 18 May 2001; received after revision 17 July 2001; accepted 24 July 2001     

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     

Zur Gestalt des Influenza-Virus

Summary Influenza-Virus particles are generally considered to be of spherical shape after conventional fixation in OsO₄. However, after fixation with KMnO₄, the elementary bodies of the Influenza-Virus (Type A Singapur I/57) — adsorbed on the surface of erythrocytes — exhibit hexagonal shape in ultrathin-sections.     

The effect of glutamate,cysteate and related amino acids on insect muscle

Summary Application of Glu, CySO₂ or CySO₃ to blowfly larvae caused paralysis and increased the membrane potential of larval muscle. The contents of Glu and CySO₃ in larval muscle containing motor nerve terminals were markedly decreased after the perfusion of high K⁺ saline solution.     

Oxygen consumption of the rat small intestine during infection with nematospiroides dubius

Summary Qo₂ of jejunal rings did not differ significantly between uninfected rats and rats infected for 7 days withNematospiroides dubius. Qo₂ of isolated jejunal mucosal epithelial cells was significantly greater 7 days after infection than in uninfected controls or at 29–36 days after infection.     

Arterial and mixed venous blood gases following DNP infusions in rabbits

Summary Hypocapnia and respiratory alkalosis was found in arterial and mixed venous blood after i.v. administration of DNP. Stimulation of ventilation resulted in steady PO₂, when PvO₂ decreased, and seemed to be independent on CO₂.     

Differences in luteinizing hormone release stimulated in rat anterior pituitary cells by leukotriene C4 and by gonadotropin-releasing hormone in vitro

Summary Continuous administration of leukotriene C₄ (LTC₄, 10^–10 M) to superfused rat anterior pituitary cells increased LH release for 40 min only, whereas in a parallel experiment gonadotropin-releasing hormone (GnRH, 10^–9 M) evoked a continuous increase in hormone secretion. In contrast to GnRH, LTC₄ did not desensitize rat anterior pituitary cells. The secretory action resulting from the administration of LTC₄ (10^–10 M) was abolished for 40 min after previous stimulation. The results documented the dual action of LTC₄ on LH exocytosis.     

The effect of hydrogen peroxide on the cyclin D expression in fibroblasts

Activation of mitogen-activated protein (MAP) kinase is essential for cyclin D1 expression and provides a link between mitogenic signalling and cell cycle progression. Hydrogen peroxide (H₂ O₂ ) activates MAP kinase; however, it is not known whether this leads to cyclin D expression. Sustained expression of cyclin D1 and D2 was observed when Her14 fibroblasts were incu-bated with 3 mM or higher H₂ O₂ concentrations. Similar results were obtained when cells were incubated in the presence of serum (FCS). However, the sustained expres-complex sion of cyclin D1 and D2 upon H₂ O₂ treatment was not due to the MAP kinase pathway, because MAP kinase kinase inhibitors did not inhibit cyclin D expression. Furthermore, cyclin D1 and D2 levels remained constant even after addition of a protein synthesis inhibitor, indicating that the effect of H₂ O₂ was not due to induction of protein synthesis. These results indicate that H₂ O₂ reversibly inhibits the ubiquitin-proteasome dependent degra-dation of cyclin D1 and D2, probably by transiently in-hibiting ubiquitination and/or the proteasome. Received 12 March 2001; received after revision 5 April 2001; accepted 9 April 2001     

IgG purification to measure the level of an iodinated thyroglobulin peptide,the 3,5,3′,5′ tetraiodo-l-tyrosyl-l-tyrosine in human serum

Summary Antibodies reacting with 3,5,3,5 tetraiodo-l-tyrosyl-l-tyrosine (I₂Tyr-I₂Tyr) were elicited in rabbits by immunization with an oxidized yeast conjugate coupled with I₂Tyr-I₂Tyr. Ion-exchange chromatography was used to purify immunoglobulins, in order to improve the specificity in measurement of I₂Tyr-I₂Tyr level in patient serum. IgG binding capacity versus I₂Tyr-I₂Tyr was considerably increased after immunoglobulin purification.Acknowledgments. The authors with to thank Mrs M. Ollier for her valuable technical assistance.     

Comparaison de l'activité respiratoire d'Esch. coli pendant la bactériolyse due aux bactériophages de la série T1 à T7

Summary The course of bacteriolysis as determinated by the oxygen consumption of the bacteria is different during bacteriolysis produced by the phagesT _1,3,5,7 as compared with that produced by phagesT _2,4,6. With phagesT _1,3,5,7, the bacteriolysis occurs, rapidly, but incompletely after a latent period. With the phagesT _2,4,6 bacteriolysis occurs slowly and the course of the respiration shows a « plateau » but goes to completion. The biochemical mechanism of the bacteriolysis therefore seems to differ in both cases.