[Modeling of cutaneous radio-contamination: effects of washings by soap and by solutions of DTPA]. / Modélisation des radiocontaminations cutanées: effets de lavages par le savon et par les solutions de DTPA.

Some neutral chirurgical soap, a solution of diethylen penta acetic acid (DTPA) at 1% and a solution at 25% of monocalcic trisodium salt of DTPA in serum have been studied on models of cutaneous contamination. Cesium 137 (137Cs) and Plutonium 239 (239Pu) have been used for the contamination. This research gives a protocol for external cutaneous decontamination which could be used in reception units for radiocontaminated wounded. Even the radioelement is unknown, DTPA at 25% represents a good solution for the treatment of general contamination, which is not the case for DTPA at 1%. A soap in which DTPA at 1% has been added is adapted for large cutaneous contamination. The treatment of ocular contamination is also studied in this article.

Immunological relevance of malonic dialdehyde (MDA): III. Immuno-enzymatic determination of human immunoglobulin binding to MDA-crosslinked proteins.

Using immunoenzymatic reactions (ELISA) with MDA-crosslinked lysozyme (ML) or poly-L-lysine (MP) and with the corresponding native protein or polypeptide, we showed that sera from normal human subjects contain immunoglobulins with antibody-like specificity for MDA-modified proteins. Inhibition studies with ML and MP showed that the chemical structures recognized by these immunoglobulins include 1-amino-3-iminopropene (AIP) bridges resulting from reactions of MDA with primary amino groups. An ELISA technique for quantitation of these immunoglobulins was developed and applied to 32 sera from healthy humans, which provided an estimation of their normal levels. Variations of these levels under pathological conditions are now being investigated.

Immunological relevance of malonic dialdehyde (MDA): II. Precipitation reactions of human sera with MDA-crosslinked proteins.

Using precipitation reactions in agarose gels (Bidimensional double diffusion: Ouchterlony. Counterimmunoelectrophoresis: CEP), we showed that: a) sera from normal human subjects contain components able to bind and precipitate with MDA-crosslinked lysozyme (ML) and not with native lysozyme, which indicates that the chemical structures involved in such bindings arise from reaction of MDA with lysozyme and probably include 1-amino-3-iminopropene (AIP) bridges. b) some if not all of these seric components are immunoglobulins. c) the F(ab')2 regions of these immunoglobulins are involved in their binding and precipitating properties. These results lead us to assume that sera from normal human subjects contain immunoglobulins with antibody-like specificity for MDA-crosslinked proteins. Nevertheless, this assumption remains to be assessed by further studies, especially about the "epitopes" involved in such reactions.

Modulation of leucocyte activation in the early phase of the rabbit burn injury.

Previous studies in burned patients have shown an early enhanced polymorphonuclear leucocyte (PMN) generating capacity for superoxide radical (O2.-), for the arachidonic acid (AA) lipoxygenase metabolite leukotriene B4 (LTB4) and for platelet activating factor-acether (PAF). These findings have been confirmed on a burn injury rabbit model. As we have suggested a pivotal role for an exaggerated initial (less than 36-48 h) neutrophil stimulation leading to a later (greater than 72 h) immuno-depression and anergy, we tried to modulate the early phase by drug therapy. A Ginkgo biloba extract (IPS200) injected i.v. in burned rabbits greatly reduced O2.- and LTB4 generation on A23187 challenge. IPS200 includes flavonoids and other polyphenols, inhibiting either arachidonic acid metabolism or PAF receptors, and may thus exert their modulating effect on PMN function in thermal injury.

Immunological relevance of malonic dialdehyde. I. Preparation of Schiff's bases from lysozyme or polylysine reacted with malonic dialdehyde.

Malonic dialdehyde (MDA) is produced in all mammalian tissues either as an end product of lipid peroxidation or as a by-product of arachidonic acid metabolism. It may either be quickly oxidized to carbon dioxide or combine covalently with primary amino groups of proteins, phospholipids or nucleic acids. In the latter case, fluorescent Schiff's bases with 1-amino-3-iminopropene (AIP) bridges are produced. MDA metabolism is now fairly well elucidated, while that of MDA-cross-linked biological molecules remains unknown. Aiming at investigating the fate of such cross-linked molecules in mammalian organisms, and their biological relevance, we tried in the present study to prepare reproducibly Schiff's bases from chicken egg white lysozyme reacted with MDA. The resulting mixture of different Schiff's bases (ML) was fractionated into single oligomeric fractions by gel-filtration chromatography. ML and the single oligomeric fractions obtained from this mixture were controlled by fluorescence measurements for their content of AIP bridges, and by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) for their content of different oligomers. ML contained monomers, dimers, trimers and other oligomers, as shown by SDS-PAGE. The corresponding single oligomeric fractions were satisfactorily separated by gel-filtration chromatography (purity better than 94%, as determined by SDS-PAGE). Schiff's bases from poly-L-lysine reacted with MDA (MP) were also prepared. Their fluorescence emission spectrum was similar to that of ML and to that of the single oligomeric fractions obtained from ML.

Radiosensitive antioxidant membrane-bound factors in rat liver microsomes: I. The roles of glutathione and vitamin E.

In vitro lipid peroxidation initiated by NADPH/ADP/Fe3+ reveals an alteration of rat liver microsomal antioxidant factors at day D+4 after whole-body gamma irradiation (8Gy). This alteration is partly reversed by GSH, and more efficiently by Trolox C, a water-soluble analog of vitamin E. This reversion by Trolox C, together with the observed 50% decrease in vitamin E content in microsomes of irradiated rats as compared to those of control animals, indicate that Trolox C acts as a free-radical scavenger like and in place of vitamin E. The antioxidant action of Trolox C is not improved in the presence of GSH, which suggests that the former acts earlier than the latter on the autoxidative free-radical chain reactions. Neither GSH, nor Trolox C, nor both antioxidants totally inhibit in vitro lipid peroxidation, which appeals attention on the possible role of extra-microsomal antioxidant factors, especially cytosolic ones.

Radiosensitive antioxidant membrane-bound factors in rat liver microsomes: II. The role of cytosol.

In vitro lipid peroxidation initiated by NADPH/ADP/Fe3+ reveals an alteration of rat liver microsomal membrane at day D+4 after whole-body gamma irradiation (8Gy). This alteration is reversed by Trolox-C 100 microM plus GSH 2mM in association with the soluble supernatant from irradiated or control rats. The antioxidant capacity of the soluble supernatant is not inactivated by gamma irradiation. We conclude that the control of in vivo lipid peroxidation is membranous and cytosolic, and that the preservation of this activity is GSH and Vitamin E dependent.

Influence of whole-body gamma-irradiation upon rat erythrocyte: lipid peroxidation and osmotic fragility.

The effects of whole-body gamma-irradiation of rats (8 Gy) on erythrocyte enzymes and biochemical components involved in lipid peroxidation were studied. Decreased superoxide dismutase and glutathione reductase activities, and lowered concentrations of reduced glutathione, were found to be the main factors responsible for the observed increase in lipid peroxidation in the erythrocytes of irradiated rats. This increased lipid peroxidation did not result in a greater tendency to hemolysis in hypotonic media; on the contrary, the mean osmotic fragility was decreased at days D + 1 and D + 3 after irradiation. The behavior of the erythrocyte populations towards hemolysis in hypotonic media appeared to be most homogeneous at days D + 4 and D + 8 after irradiation, which correspond to maxima of malonic dialdehyde concentrations in erythrocytes. Such a synchrony of variations suggests that crosslinking of primary amino groups of proteins or phospholipids by malonic dialdehyde might produce a rigidification in erythrocyte membranes, possibly leading to a more homogeneous behavior of the erythrocyte populations towards hemolysis in hypotonic media.

Influence of whole-body gamma irradiation upon arachidonic acid metabolism in rat platelets.

The effects of whole-body gamma irradiation (8.4 Gy) were studied on arachidonic acid (AA) metabolism in rats' blood platelets, from day D + 1 to day D + 10 after irradiation. AA conversion into thromboxane B2 (TxB2) increased at D + 1 and then gradually decreased to very low values from D + 7 to D + 10. This decrease in the conversion of exogenous AA into TxB2 was due to a lower AA incorporation into platelets and not to a decrease of cyclooxygenase and thromboxane-synthetase activities. AA incorporation into membrane phospholipids of blood platelets was much more decreased than AA incorporation into whole platelets; moreover, the lipid composition of the platelet membranes was markedly modified after irradiation, which must have resulted in structural and functional changes in these membranes; from these effects of whole-body gamma irradiation on platelets, the latter's membranes appeared as a major site of in vivo radiation damage in these cells.

Leukocytic functions in burn-injured patients.

Anergy associated with an increase in suppressor helper T cell (Tc) ratio and a decrease in natural killer (NK) is one main cause of death following thermal injury (Tl). Recently, in vitro studies have shown that LTB4 can induce human Tc to exert suppressor cell activity, and incubation of lymphocytes with LTB4 for 24 hours significantly suppressed NK cell activity. Thus, we undertook an investigation of both AA metabolism and immunologic response in 20 patients who suffered 40-90% total body surface area (TBSA) burns. Cyclooxygenase (CO:RIA) and lipoxygenase (LO;HPLC det.) metabolites and superoxide (O2 X-) production were measured in stimulated polymorphonuclear cells (PMNL) (A 23187 +/- AA for icosanoid release; phorbol myristate acetate for O2 X-production). Lyso-paf-acether (P-LPA) was measured in plasma samples. Ca2+-dependent K+permeability in PMNL was measured by the cell K+ release induced by A 23187. Tc and Tc subsets were determined using monoclonal antibodies (OKT3+, OKT4+ and OKT8+). A biphasic sequential release of the different substances (leukocytic icosanoids and O2 X-) was monitored: increase (approximately 36-48 h after Tl) and decrease (greater than or equal to 72 h after Tl). The increase in AA stimulation was more transient than that of O2 X -. The decline in the release of AA metabolites and O2 X-production was associated with the anergic phase (decrease OKT4+/OKT8+ ratio) and with the clinical outcome of the patients. The decrease in LTB4 and other LO metabolites could explain the impairment of neutrophil chemotaxis. Ca2+-dependent K+ permeability increased early up to 2 or 3 times normal. In order to go further with the mechanism of inhibition of LTB4 and O X-release, the effect of Tl plasma was assayed on normal leukocytes: a 10 min incubation with such plasma was sufficient to abolish LTB4 secretion. A less important inhibition was observed with O2 X-release (-32%) and Ca2+-dependent K+ permeability (-30%). Plasma inhibition seems to be due to a thermolabile factor(s) [protein(s): "suppressive factor(s) of membrane activation "SFMA] which is (are) under active investigation using gel-filtration chromatography and fast protein liquid chromatography (FPLC). Among the SFMAs, certain acute phase proteins could play a key role: i.e., incubation (10 min) of normal PMNL with ceruloplasmin (1 mg/ml) abolished LO products and O X 2-release.

[Effect of irradiation on the in vivo and in vitro subcellular distribution of radiostrontium in rat liver]. / Effet de l'irradiation sur la distribution subcellulaire du radiostrontium dans le foie de rat in vivo et in vitro.

Hepatic mitochondrial strontium uptake is enhanced in whole body irradiated rats. Effect is observed on animals but not on irradiated organ slices. It is likely that a hypophyso - adrenal axis hormone plays a role in changes observed in vivo.