Direct Evidence of Free Radical Production after Ischaemia and Reperfusion and Protective Effect of Desferrioxamine: Esr and Vitamin E Studies

in European Journal of Vascular Surgery (1994), 8(5), 537-43

After surgical renal revascularisation, warm renal ischaemia due to renal artery cross-clamping contributes to postoperative renal dysfunction. After reperfusion, free radicals are thought to be a ... [more ▼]

After surgical renal revascularisation, warm renal ischaemia due to renal artery cross-clamping contributes to postoperative renal dysfunction. After reperfusion, free radicals are thought to be a significant cause of injury. Nevertheless, indisputable proof of free radical production is scarce, partly because of their transient nature. In this study, electron paramagnetic resonance and vitamin E levels were used to demonstrate the free radical production after renal ischaemia and reperfusion. Rabbit kidneys were submitted either to 15 or 60 minutes of ischaemia followed by reperfusion. A spin trap agent (alpha-phenyl-N-tert-butyl nitrone (PBN), 20mg/ml, 1 ml/min) was infused during reperfusion directly into the left renal artery via an aortic catheter before declamping. Blood samples were selectively drawn from the left renal vein for ESR analysis (Varian spectrometer E109) of lipidic residues extracted from blood samples. The vitamin E content of the left renal cortex was determined by HPLC procedure. The right renal cortex was used as a control for the vitamin E values. In the venous effluent, ESR analysis revealed the formation of a spectrum consisting of a triplet of asymmetric doublets. This signal resulted from the spin trapping by PBN of a mixture of both oxygen- and carbon- centred lipidic radicals. The amplitude of the signal which is proportional to the amount of free radicals was significantly higher after 60 minutes ischaemia than after 15 minutes.(ABSTRACT TRUNCATED AT 250 WORDS) [less ▲]

Evidence for Free Radical Formation During Human Kidney Transplantation

in Free Radical Biology & Medicine (1993), 15(3), 343-8

Fourteen patients undergoing kidney transplantation were studied for evidence of the production of free radicals as assessed by the measurement of vitamin E (an index of lipid peroxidation) and of ... [more ▼]

Fourteen patients undergoing kidney transplantation were studied for evidence of the production of free radicals as assessed by the measurement of vitamin E (an index of lipid peroxidation) and of myeloperoxidase (a marker of neutrophil activation) in the systemic blood. Early (2 min) and late revascularization (30 min) of the kidney were respectively associated with a significant decrease of 35.5 and 40% of the initial level of plasma vitamin E. This consumption paralleled to the decrease of the vitamin E/total lipids ratio, a better indicator of vitamin E status. Heparin administration preceding renal artery clamping resulted in a twofold significant increase of baseline plasma myeloperoxidase (MPO) level (523 +/- 214 ng/ml). At kidney reperfusion, MPO concentration rose again and reached a maximum value of 1,653 +/- 882 ng/ml, indicating the presence of considerable neutrophil activation. A return to the baseline value was observed after 30 min of reperfusion. A short discussion about the possible origin of this MPO increase is given. Taken together, these data strongly suggest that free radical production, leading to lipid peroxidation phenomena, can occur within the early phase of kidney revascularization. Preliminary data using electron spin resonance with the spin-trapping technique strengthen this hypothesis. [less ▲]

Ischémie - reperfusion rénale: des altérations biochimiques aux lésions anatomo-pathologiques et aux troubles fonctionnels

in Kieffer (Ed.) Chirurgie de l'artère rénale (1993)

Plasma myeloperoxidase and vitamin E levels in head injury: preliminary results related to outcome.

in Journal of Neurosurgical Anesthesiology (1992), 4(1), 26-30

This preliminary study was designed to assess a possible role of neutrophil activation and to determine the prognostic value of plasma myeloperoxidase (MPO) and vitamin E (Vit. E) levels in severe head ... [more ▼]

This preliminary study was designed to assess a possible role of neutrophil activation and to determine the prognostic value of plasma myeloperoxidase (MPO) and vitamin E (Vit. E) levels in severe head injury. Plasma MPO and Vit. E levels were measured in nine severely head-injured patients (Glasgow Coma Score </=8) (ages 12-80 years) 6, 12. 18, 24, and 30 h after trauma. Patients were classified into two groups according to outcome after discharge from the ICU: group D (death; n = 5) and group S (survival; n = 4). Plasma MPO levels were increased immediately after trauma and then decreased. The MPO peak observed after 6 h was significantly higher (p < 0.05) in group D (mean +/- SEM: 1,237 +/- 122 ng/ml) than in group S (mean +/- SEM: 543 +/- 148 ng/ml). Plasma Vit. E levels were lower than normal values and decreased over time. They were always significantly lower (p < 0.05) in group D than in group S, except for the first sample. These differences cannot be explained entirely by total plasma lipid (TL) values since no statistical difference in TL concentrations was found between the two groups during the course of study. The ratio of Vit. E to TL. considered as the best index of Vit. E status, was lower in group D than in group S. and the difference reached statistical significance (p < 0.05) 12 h after trauma. In conclusion, in spite of the limited number of patients included in this study, it appears that severe head injury is associated with an increase in MPO and a decrease in Vit. E levels. These biochemical changes are of greater magnitude in group D than in group S; they suggest neutrophil activation and lipoperoxidation processes. Finally, plasma MPO and Vit. E seem to be new discriminant factors of outcome in head-injured patients. [less ▲]

Self-Labeling of Human Polymorphonuclear Leucocyte Myeloperoxidase with 125iodine

in Experientia (1991), 47(9), 952-7

In order to obtain a radioimmunoassay (RIA) technique for the measurement of human plasma myeloperoxidase (MPO), we purified the enzyme from polymorphonuclear granulocytes (neutrophils), and compared ... [more ▼]

In order to obtain a radioimmunoassay (RIA) technique for the measurement of human plasma myeloperoxidase (MPO), we purified the enzyme from polymorphonuclear granulocytes (neutrophils), and compared three methods of labeling it with 125Iodine:chloramine T, lactoperoxidase, and an original technique of 'self labeling' based on the ability of the enzyme to oxidize and bind 125I in the presence of H2O2. The chloramine T technique produced a degraded protein, as well shown by a high non-specific binding of tracer to antibody. The lactoperoxidase technique did not succeed in labeling MPO with an adequate specific activity. In contrast, the self-labeling method gave a stable tracer with a specific activity of 23 microCi/micrograms MPO (85 MBq), a satisfactory level of immunoreactivity, and a low-specific binding (less than or equal to 3%). After labeling, purification of tracer was achieved by gel filtration chromatography in phosphate buffer (0.05 M; pH7) to which 0.1% poly-L-lysine was added. The labeled molecule remained stable for 40 days and could be used for RIA with a polyclonal antibody raised in rabbits. [less ▲]

Fast double antibody radioimmunoassay of human granulocyte myeloperoxidase and its application to plasma.

in Journal of Immunological Methods (1991), 137(2), 181-191

The haem enzyme myeloperoxidase (MPO) (EC 1.11.1.7) with a spectral A430/A280 ratio greater than 0.7 and a specific activity of 125 U/mg was purified from isolated human neutrophils. To obtain a ... [more ▼]

The haem enzyme myeloperoxidase (MPO) (EC 1.11.1.7) with a spectral A430/A280 ratio greater than 0.7 and a specific activity of 125 U/mg was purified from isolated human neutrophils. To obtain a radioimmunoassay (RIA) for this enzyme, a specific antiserum against human neutrophil MPO was raised in rabbits and used at an initial dilution of 1/10,000. MPO labelled with 125iodine by a technique of self-labelling in the presence of H2O2, had a specific activity of 24 mCi/mg. After incubation at room temperature (2 h) and separation by double antibody precipitation in the presence of polyethylene glycol, the sensitivity of the RIA was 21 ng/ml. The RIA showed good precision and accuracy with intra- and interassay coefficients of variation of less than 7% for MPO concentrations ranging from 100 to 800 ng/ml, and satisfactory recoveries of known amounts of exogenous MPO in plasma. For the measurement of MPO in blood, the best sampling technique was to collect blood into EDTA. Rapid centrifugation (within 20 min) was necessary for blood collected into heparin. Mean MPO values in normal individuals were 340 +/- 98 ng/ml in EDTA plasma (n = 152) and 332 +/- 82 ng/ml in heparinized plasma (n = 34). When MPO was measured 12-6 h after injury in critically ill patients high values (above 1000 ng/ml) were found in 6/15 patients with multiple injuries. In patients with sepsis (n = 22), MPO values were always above 1000 ng/ml. [less ▲]

Activation of human immunodeficiency virus type 1 by an oxidative stress

in AIDS Research and Human Retroviruses (1990)

Equine postanaesthetic myositis: thromboxanes, prostacyclin and prostaglandin E2 production

in Veterinary Research Communications (1988), 12

1019-Pentane measurement in man as an index of lipoperoxidation

in Bioelectrochemistry and Bioenergetics (Lausanne, Switzerland) (1987), 18

La troxérutine et la rutine sulfate sodique possèdent-elles encore les propriétés de la rutine?

in Annales Pharmaceutiques Françaises (1987), 45(4), 329-334

Rutin, sodium rutin sulfate and troxerutin were tested for their antiradicalar, antilipoperoxidant, antioxygen properties and for their activity on in vivo prostaglandin biosynthesis.In each test, rutin ... [more ▼]

Rutin, sodium rutin sulfate and troxerutin were tested for their antiradicalar, antilipoperoxidant, antioxygen properties and for their activity on in vivo prostaglandin biosynthesis.In each test, rutin is more powerful than its derivatives. This fact results mainly on a free orthodiphenolic group ( 3'-4'-hydroxyls on ring B . [less ▲]