摘要:

A large number of publications recently have drawn strong analogies between the production of active oxygen species in plant cells and the“oxidative burst”of the phagocyte, even to the point of constructing elaborate models involving receptor mediated G-protein activation of a plasmalemma NADPH oxidase in plant cells. However there are potentially other active oxygen species generating systems at the plant cell surface. The present work examines these alternatives with particular emphasis on the rapid production of active oxygen species, in common with a number of other systems, by suspension-cultured cells of French bean on exposure to an elicitor preparation from the fungal pathogenColletotrichum lindemuthianum. The cells show a rapid increase in oxygen uptake which is followed shortly afterwards by the appearance of a burst of these active oxygen species, as measured by a luminescence assay, which is probably all accounted for by hydrogen peroxide. An essential factor in this production of H2O2appears to be a transient alkalinization of the apoplast where the pH rises to 7.0-7.2. Dissipation of this pH change with a number of treatments, including ionophores and strong buffers, substantially inhibits the oxidative burst. Little evidence was found for enhanced activation of a membrane-bound NADPH oxidase. However the production of H2O2under alkaline conditions can be modelledin vitrowith a number of peroxidases, one of which, an Mr46,000 wall-bound cationic peroxidase, is able to sustain H2O2production at neutral pH unlike the other peroxidases which only show low levels of this reaction under such conditions and have pH optima at values greater than 8.0. On the basis of such comparative pH profiles between the cells and the purified peroxidase and further inhibition studies a direct production of H2O2from the wall peroxidase in French bean cells is proposed. These experiments may mimic some of the responses to plant pathogens, particularly the hypersensitive response, which is an important feature of resistance. A cell wall peroxidase-origin for the oxidative burst is clearly different from a model consisting of receptor activation of a plasmaiemma-localised NADPH oxidase generating superoxide. An alternative simple and rapid mechanism thus exists for the generation of H2O2which does not require such multiple proteinaceous components.

ISSN:1071-5762    

DOI:10.3109/10715769509065273

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Removal of adventitious redox-active metals from buffers by treatment with Chelex resin is a widely used procedure in free radical research. Use of a new batch of Chelex-100 resin in our laboratory coincided with a sudden inability to oxidise low-density lipoprotein with copper. We found that copper-mediated oxidation of ascorbate in water treated with the same batch of Chelex was inhibited when compared with untreated water and water treated with a different batch of the resin. Washing the Chelex removed the inhibitory effect suggesting that material was leaching from the resin. The washing procedure for Chelex-100 described is simple and can be scaled up. Oxidation of ascorbate with low concentrations of copper can be used to test the quality of batches of the resin.

ISSN:1071-5762    

DOI:10.3109/10715769509065274

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Peroxynitrite anion is a powerful oxidant which can initiate nitration and hydroxylation of aromatic rings. Peroxynitrite can be formed in several ways, e.g. from the reaction of nitric oxide with superoxide or from hydrogen peroxide and nitrite at acidic pH. We investigated pH dependent nitration and hydroxylation resulting from the reaction of hydrogen peroxide and nitrite to determine if this reaction proceeds at pH values which are known to occurin vivo. Nitration and hydroxylation products of tyrosine and salicylic acid were separated with an HPLC column and measured using ultraviolet and electrochemical detectors. These studies revealed that this reaction favored hydroxylation between pH 2 and pH4, while nitration was predominant between pH 5 and pH 6. Peroxynitrite is presumed to be an intermediate in this reaction as the hydroxylation and nitration profiles of authentic peroxynitrite showed similar pH dependence. These findings indicate that hydrogen peroxide and nitrite interact at hydrogen ion concentrations present under some physiologic conditions. This interaction can initiate nitration and hydroxylation of aromatic molecules such as tyrosine residues and may thereby contribute to the biochemical and toxic effects of the molecules.

ISSN:1071-5762    

DOI:10.3109/10715769509065275

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Estimations ofα-tocopherol content were made on a series of human necropsy samples of normal arterial wall and of atherosclerotic lesions. The results were compared with stage of lesion, shown by histology, and with the amounts of cholesterol and hydroxycholesterols in the same lesions. The ratio ofα-tocopherol to cholesterol levels varied widely in normal arterial wall but was consistently low in lesions, especially in lesions rich in macrophage foam cells. The results suggested that significant accumulation of hydroxycholesterols, found almost exclusively in lesions, only occurred whenα-tocopherol levels were low in relation to the cholesterol content. This suggests that oxidative activity in the lesion may lead to significant oxidation of constituents of low-density lipoprotein only afterα-tocopherol has been depleted.

ISSN:1071-5762    

DOI:10.3109/10715769509065276

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Hemopexin, a heme-binding serum glycoprotein, is thought to play an important role in the prevention of oxidative damage that may be catalysed by free heme. Through the use of EPR techniques, the generation of free radicals from organic hydroperoxides by heme and heme-hemopexin complexes, and the concomitant formation of high oxidation-state iron species has been studied; these species are implicated as causative agents in processes such as cardiovascular disease and carcinogenesis. From the rates of production of these species from both n-alkyl and branched hydroperoxides, it has been inferred that the dramatic reduction in the yield of oxidising species generated by heme upon its complexation with hemopexin arises from steric hindrance of the access of hydroperoxide to the bound heme.

ISSN:1071-5762    

DOI:10.3109/10715769509065277

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Performing the deoxyribose (DR) assay for determination of the rate constants for reaction of non steroidal antiinflammatory drugs with hydroxyl radicals led to some unusual competition plots. The molecules from the arylpropionic family of drugs: ibuprofen, flurbiprofen, ketoprofen and naproxen produced the linear relationship. However, acemetacin, diclofenac Na, flufenamic acid, indometacin, niflumic acid, tolmetin Na and sulindac presented non linear competition plots manifesting at relatively low drug concentrations. This effect was corrected by increasing DR concentrations from 2.8 mM to 15 mM. The modification did not affect rate constants values for those derivatives which already presented a linear plot at 2.8 mM, but allowed to calculate rate constants for other compounds. It is suggested that the experimental conditions have to be adapted particularly for those derivatives with a relatively high. rate constant for reaction with the radical species. The oxicam derivatives (tenoxicam and piroxicam) presented another kind of deviation that revealed a prooxidant effect in this system: non linear plots were also observed at relatively low drug concentrations, but in the opposite direction than for the other molecules. This last effect was independent of DR concentration but could be corrected by increasing ascorbate concentration in the system.

ISSN:1071-5762    

DOI:10.3109/10715769509065278

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

U937 is a monocytic cell line dependent on low density lipoprotein (LDL) receptor-mediated uptake of cholesterol for proliferation. However, exposure of U937 cells to LDL also results in an oxidative modification of LDL. We report here that the oxidative modification of LDL by U937 cells results in inhibition of growth and cell death. This finding suggests that analysis of U937 cell growth in presence of LDL may be used to determine the susceptibility of LDL to biological oxidative modification. There was an inverse association between the effect of LDL on U937 cell growth and the rate of degradation of U937 cell-modified LDL in mouse peritoneal macrophages (r=-0.82, p<0.05) suggesting a coupling between proneness of LDL to develop cytotoxicity and affinity for scavenger receptors. In a group of young post-infarction patients (n=18) the susceptibility of LDL to chemical oxidation as determined by analysis of the lag phase for formation of conjugated diens in presence of copper ions was compared with the biological modification of LDL as assessed by analysis of U937 cell growth in presence of LDL. The results demonstrated a close relation between the estimates of chemical oxidation and biological modification (r=0.86, p<0.005) suggesting that LDL, which is prone to become oxidised by copper also is more prone to become modified by cells in vivo.

ISSN:1071-5762    

DOI:10.3109/10715769509065279

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

The potential for free radical release has been measured by means of the spin trapping technique on three kinds of iron containing particulate: two asbestos fibers (chrysotile and crocidolite); an iron-exchanged zeolite and two iron oxides (magnetite and haematite). DMPO (5,5′-dimethyl-1 -pirroline-N-oxide), used as spin trap in aqueous suspensions of the solids, reveals the presence of the hydroxyl and carboxylate radicals giving rise respectively to the two adducts [DMPO-OH] and [DMPO-CO2], each characterized by a well-defined EPR spectrum. Two target molecules have been considered: the formate ion to evidence potential for hydrogen abstraction in any biological compartment and hydrogen peroxide, always present in the phagosome during phagocytosis. The kinetics of decomposition of hydrogen peroxide has also been measured on all solids. Ferrozine and desferrioxamine, specific chelators of Fe(II) and Fe(III) respectively, have been used to remove selectively iron ions. Iron is implicated in free radical release but the amount of iron at the surface is unrelated to the amount of radicals formed. Only few surface ions in a particular redox and coordination state are active. Three different kinds of sites have been evidenced: one acting as H abstractor, the other as a heterogeneous catalyst for hydroxyl radical release, the third one related to catalysis of hydrogen peroxide disproportionation. In both mechanisms of free radical release, the Fe-exchanged zeolite mimics the behaviour of asbestos whereas the two oxides are mostly inert. Conversely magnetite turns out to be an excellent catalyst for hydrogen peroxide disproportionation while haematite is inactive also in this reaction. The results agree with the implication of a radicalic mechanism in thein vitroDNA damage and in thein vivotoxicity of asbestos.

ISSN:1071-5762    

DOI:10.3109/10715769509065280

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Our earlier studiesin vitrohave shown that eugenol inhibits liver microsomal monooxygenase activities and carbon tetrachloride (CCl4)-induced lipid peroxidation (Free Rad. Res. 20,253-266,1994). The objective of the present investigation was to study thein vivoprotective effect of eugenol against CCI4toxicity. Eugenol (5 or 25 mg/kg body wt) given orally for 3 consecutive days did not alter the levels of serum glutamic oxalacetic transaminase (SGOTJ, microsomal enzymes such as cytochrome P450 reductase, glucose-6-phosphatase (G-6-Pase) xenobiotic-metabolizing enzymes (aminopyrine-N-demethylase, N-nitrosodimethylamine-demethylase and ethoxyresorufin-O-deethylase) and liver histology. Doses of eugenol (5 or 25 mg/kg) administered intragastrically to each rat on three consecutive days i.e. 48 hr, 24 hr and 30 min before a single oral dose of CCU (2.5 ml/kg body wt) prevented the rise in SGOT level without appreciable improvement in morphological changes in liver. Eugenol pretreatment also did not influence the decrease in microsomal cytochrome P450content, G-6-Pase and xenobiotic-metabolizing enzymes brought about by CCI4. Since eugenol is metabolized and cleared rapidly from the body, the dose schedule was modified in another experiment. Eugenol (0.2,1.0,5.0 or 25 mg/kg) when given thrice orally i.e. prior to (-1 hr) along with (0 hr) and after (+ 3 hr) the i.p. administration of CCI4(0.4 ml/kg) prevented significantly the rise in SGOT activity as well as liver necrosis. The protective effect was more evident at 1 mg and 5 mg eugenol doses. However, the decrease in microsomal G-6-Pase activity by CCI4treatment was not prevented by eugenol suggesting that the damage to endoplasmic reticulum is not protected. The protective effect of eugenol against CC14induced hepatotoxicity is more evident when it is given concurrently or soon after rather than much before CCU treatment.

ISSN:1071-5762    

DOI:10.3109/10715769509065281

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

ISSN:1071-5762    

DOI:10.3109/10715769509065282

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor