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Biomedical subjects

S D Aust

Publications and source records attributed to S D Aust.

At least 145 records · Page 8Linked to original sources
PubMed1988

Reconstituted microsomal lipid peroxidation: ADP-Fe3+-dependent peroxidation of phospholipid vesicles containing NADPH-cytochrome P450 reductase and cytochrome P450.

A reconstituted lipid peroxidation system consisting of rat liver microsomal NADPH-cytochrome P450 reductase and cytochrome P450 incorporated into phospholipid vesicles was developed and characterized. Peroxidation of the vesicles required NADPH and ADP-Fe3+, just as in the NADPH-dependent peroxidation of microsomes. The peroxidation of the vesicles was inhibited 30-50% by superoxide dismutase, depending upon their cytochrome P450 content: those with higher cytochrome P450 contents exhibited greater rates of malondialdehyde formation which were less sensitive to inhibition by superoxide dismutase. When cytochrome P450 was incorporated into vesicles, EDTA-Fe3+ was not required for lipid peroxidation, distinguishing this system from the one previously described by Pederson and Aust [Biochem. Biophys. Res. Comm. 48, 789; 1972]. Since at least 50% of the malondialdehyde formation in the vesicular system was not inhibited by superoxide dismutase, alternative means of iron reduction (O2-.-independent) were examined. It was found that rat liver microsomes or a reconstituted mixed function oxidase system consisting of NADPH-cytochrome P450 reductase and cytochrome P450 in dilauroylphosphatidylcholine micelles reduced ADP-Fe3+ under anaerobic conditions.

Adenosine Diphosphate↗
PubMed1988

Inhibition of 2-aminofluorene mutagenesis in bacteria by inducers of cytochrome P-450d.

Certain chemical inducers of rat liver microsomal cytochrome P-450d are tightly bound to the cytochrome. We investigated the ability of two inducers of cytochrome P-450d, Aroclor 1254 and isosafrole, to inhibit the microsomal activation of 2-aminofluorene to a mutagen as measured in Salmonella typhimurium. Butanol treatment of microsomes from isosafrole-treated rats removed an inhibitory metabolite of isosafrole and increased 2-aminofluorene mutagenesis by approximately 2-fold over controls. Butanol treatment of microsomes from Aroclor 1254-treated rats failed to either remove any of the Aroclor 1254 associated with microsomal cytochrome P-450 or affect 2-aminofluorene-induced mutagenesis. However, addition of Aroclor 1254 to butanol-treated microsomes from isosafrole-treated rats almost completely inhibited 2-aminofluorene mutagenesis. Aroclor 1254 completely inhibited the cytochrome P-450d-dependent estradiol 2-hydroxylase activity of butanol-treated microsomes from isosafrole-treated rats. Thus, we suspect that certain congeners from Aroclor 1254, a widely used mixture for induction of cytochrome P-450 activities, could inhibit cytochrome P-450d and partially mask its ability to metabolize some chemicals to mutagens.

Animals↗
PubMed1988

Biodegradation of pentachlorophenol by the white rot fungus Phanerochaete chrysosporium.

Extensive biodegradation of pentachlorophenol (PCP) by the white rot fungus Phanerochaete chrysosporium was demonstrated by the disappearance and mineralization of [14C]PCP in nutrient nitrogen-limited culture. Mass balance analyses demonstrated the formation of water-soluble metabolites of [14C]PCP during degradation. Involvement of the lignin-degrading system of this fungus was suggested by the fact the time of onset, time course, and eventual decline in the rate of PCP mineralization were similar to those observed for [14C]lignin degradation. Also, a purified ligninase was shown to be able to catalyze the initial oxidation of PCP. Although biodegradation of PCP was decreased in nutrient nitrogen-sufficient (i.e., nonligninolytic) cultures of P. chrysosporium, substantial biodegradation of PCP did occur, suggesting that in addition to the lignin-degrading system, another degradation system may also be responsible for some of the PCP degradation observed. Toxicity studies showed that PCP concentrations above 4 mg/liter (15 microM) prevented growth when fungal cultures were initiated by inoculation with spores. The lethal effects of PCP could, however, be circumvented by allowing the fungus to establish a mycelial mat before adding PCP. With this procedure, the fungus was able to grow and mineralize [14C]PCP at concentrations as high as 500 mg/liter (1.9 mM).

Biodegradation, Environmental↗
PubMed1987

Superoxide-dependent redox cycling of citrate-Fe3+: evidence for a superoxide dismutaselike activity.

Citrate-Fe3+, reportedly a physiological chelate, exhibits superoxide dismutaselike activity, as evidenced by the inhibition of xanthine oxidase-dependent cytochrome c reduction; the dismutation of xanthine oxidase-generated superoxide to hydrogen peroxide and oxygen, and the enhanced disproportionation of potassium superoxide. The catalytic activity of citrate-Fe3+ corresponds, on a molar basis, to 0.03% of that of copper- and zinc-containing superoxide dismutase. Although weak, this activity enables citrate-Fe3+ to inhibit superoxide and ADP-Fe3+ -dependent peroxidation of extracted microsomal lipids. Also, the dismutase activity of citrate-Fe3+ interferes with its ability to promote lipid peroxidation. It is proposed that chelation of Fe3+ by citrate may represent a protective mechanism against the deleterious consequences of superoxide generation.

Chelating Agents↗
PubMed1987

The requirement for iron (III) in the initiation of lipid peroxidation by iron (II) and hydrogen peroxide.

The initiation of lipid peroxidation by Fe2+ and H2O2 (Fenton's reagent) is often proposed to be mediated by the highly reactive hydroxyl radical. Using Fe2+, H2O2, and phospholipid liposomes as a model system, we have found that lipid peroxidation, as assessed by malondialdehyde formation, is not initiated by the hydroxyl radical, but rather requires Fe3+ and Fe2+. EPR spin trapping with 5,5-dimethyl-1-pyrroline-N-oxide and the bleaching of para-nitrosodimethylaniline confirmed the generation of the hydroxyl radical in this system. Accordingly, catalase and the hydroxyl radical scavengers mannitol and benzoate efficiently inhibited the generation and the detection of hydroxyl radical. However, catalase, mannitol, and benzoate could either stimulate or inhibit lipid peroxidation. These unusual effects were found to be consistent with their ability to modulate the extent of Fe2+ oxidation by H2O2 and demonstrated that lipid peroxidation depends on the Fe3+:Fe2+ ratio, maximal initial rates occurring at 1:1. These studies suggest that the initiation of liposomal peroxidation by Fe2+ and H2O2 is mediated by an oxidant which requires both Fe3+ and Fe2+ and that the rate of the reaction is determined by the absolute Fe3+:Fe2+ ratio.

Animals↗
PubMed1987

Relationship of basic research in toxicology to environmental standard setting: the case of polybrominated biphenyls in Michigan.

The accidental contamination of dairy cattle feed in Michigan in 1973-74 with polybrominated biphenyls (PBB) led to the contamination of cattle and people consuming their products. This led to an extensive animal and product monitoring and disposal program conducted by the Michigan Department of Agriculture and the Department of Natural Resources. It also led to several studies of the people of Michigan, extensive research on the chemicals, and an unprecedented establishment by the Legislature of a Toxic Substance Control Commission. Only a few relatively minor components of the PBB mixture that contaminated Michigan are metabolized and another group of minor components seem responsible for the toxicity, which, similar to that caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), includes induction of microsomal enzymes, liver hypertrophy, thymic involution, porphyria, anorexia and chloracne. PBB were found to produce the "dioxin-like" toxicity with roughly 0.01% the potency of TCDD. Both non-toxic as well as toxic congeners were found to be tumor promotors. To date it is impossible to unequivocally conclude that any human health effects can be attributed to PBB. The Toxic Substance Control Commission was established as an independent oversight body with responsibility to gather information, investigate, coordinate and make recommendations concerning toxic substances and the handling of toxic substances incidents. The Commission has declared two toxic substances emergencies and made several recommendations for regulating and solving toxic substances problems but its major activities have evolved towards a role as an environmental ombudsman.

Animals↗
PubMed1987

The effect of 3,4,3',4'-tetrachlorobiphenyl on plasma retinol and hepatic retinyl palmitate hydrolase activity in female Sprague-Dawley rats.

The Effect of 3,4,3',4'-Tetrachlorobiphenyl on Plasma Retinol and Hepatic Retinyl Palmitate Hydrolase Activity in Female Sprague-Dawley Rats. Powers, R.H., Gilbert, L.C., and Aust, S.D. (1987). Toxicol Appl. Pharmacol. 89, 370-377. A single ip dose of 1, 5, or 15 mg/kg 3,4,3',4'-tetrachlorobiphenyl (TCB) caused a dose-dependent depression of plasma retinol levels 24 hr after treatment of female Sprague-Dawley rats. The loss of plasma retinol appeared to be a function of depressed levels of the retinol-retinol-binding protein (RBP)-transthyretin ternary complex. No free retinol-RBP was observed in plasma from treated animals. Hepatic retinyl palmitate hydrolase (RPH) activity was also depressed and highly and positively correlated to the plasma retinol levels. TCB was determined to be a noncompetitive inhibitor of partially purified RPH with a KI of 91 microM. Incubation of TCB with liver microsomes and NADPH decreased the inhibition of RPH. Doses of either 2,4,5,2',4',5'-hexachlorobiphenyl (HCB) or 3,4,5,3',4',5'-HCB equimolar to the 15 mg/kg TCB dose failed to cause a similar depression of plasma retinol in treated female rats. We conclude that, unlike other polychlorinated biphenyl congeners, TCB causes a depression of plasma retinol by inhibition of hepatic RPH.

Animals↗
PubMed1987

Specific binding of polyhalogenated aromatic hydrocarbon inducers of cytochrome P-450d to the cytochrome and inhibition of its estradiol 2-hydroxylase activity.

Treatment of male Sprague-Dawley rats with 3,4,5,3',4',5'-hexabromobiphenyl (HBB) at 10 mumol/kg followed by purification of hepatic microsomal cytochrome P-450d revealed that HBB remained specifically bound to P-450d throughout purification. Binding was noncovalent since HBB was removed by extraction with dichloromethane. Although HBB induced both cytochrome P-450c and P-450d, specific immunoprecipitation of these isozymes from HBB-treated rats showed that HBB was associated only with cytochrome P-450d. Quantitation of HBB and cytochrome P-450d in microsomes from HBB-treated rats suggested a 0.9:1 ratio of HBB to cytochrome P-450d. Five other halogenated aromatic hydrocarbon inducers of cytochrome P-450d, bearing steric similarity to HBB (including 2,3,7,8-tetrachlorodibenzo-p-dioxin), were associated with cytochrome P-450d when used to induce cytochrome P-450d in rats. HBB inhibited estradiol 2-hydroxylase activity of purified cytochrome P-450d in a noncompetitive manner with an I50 of 38 nM for 50 nM P-450d whereas its noncoplanar isomer, 2,4,5,2',4',5'-hexabromobiphenyl, had an I50 over 700-fold higher. Thus certain polyhalogenated aromatic hydrocarbons, with the capacity to induce cytochrome P-450d also bind to the cytochrome when used as inducing agents and inhibit catalytic activity of the cytochrome.

Animals↗
PubMed1987

Brain cortex tissue Ca, Mg, Fe, Na, and K following resuscitation from cardiac arrest in dogs.

Recent evidence suggests that ultimate neurologic injury following cardiac arrest and resuscitation may be largely determined by biochemical events occurring during reperfusion. To test this hypothesis and further characterize the time course of some of these events, we examined tissue samples from the parietal cortex for their total content of calcium (Ca), magnesium (Mg), iron (Fe), sodium (Na), and potassium (K) after 10 minutes, two hours, four hours, and eight hours of reperfusion following a 15-minute cardiac arrest in dogs. After 10 minutes of reperfusion, there were relatively small, but significant, increases in the total tissue content of Ca and Na, as compared to nonischemic controls. All values had returned to normal at two hours and remained normal at four hours of reperfusion. However, at eight hours of reperfusion, Ca and Na content approximately doubled and K content was reduced by half. There were no significant changes at any time in the tissue content of Fe or Mg. We conclude that with between four and eight hours of reperfusion following a 15-minute cardiac arrest, a major defect occurs in many cells of the cortex with respect to their ability to control ionic balances of Ca, Na, and K. This might be explained either by failure of the energy-dependent ionic pumps or by more generalized damage to the membrane permeability barrier by a process such as lipid peroxidation.

Animals↗
PubMed1987

An investigation into the mechanism of citrate-Fe2+-dependent lipid peroxidation.

Chelation by citrate was found to promote the autoxidation of Fe2+, measured as the disappearance of 1,10-phenanthroline-chelatable Fe2+. The autoxidation of citrate-Fe2+ could in turn promote the peroxidation of microsomal phospholipid liposomes, as judged by malondialdehyde formation. At low citrate-Fe2+ ratios the autoxidation of Fe2+ was slow and the formation of malondialdehyde was preceded by a lag phase. The lag phase was eliminated by increasing the citrate-Fe2+ ratio, which also increased the rate of Fe2+ autoxidation. The Fe2+ autoxidation product required for the initiation of lipid peroxidation was characterized as being Fe3+. As direct evidence of this, linear initial rates of lipid peroxidation were obtained via the combination of citrate-Fe2+ and citrate-Fe3+, optimum activity occurring at a Fe3+-Fe2+ ratio of 1:1. Evidence is also presented to suggest that the superoxide and the hydrogen peroxide that are formed during the autoxidation of citrate-Fe2+ can either stimulate or inhibit lipid peroxidation by affecting the yield of citrate-Fe3+ from citrate-Fe2+. No evidence was obtained for the participation of the hydroxyl radical in the initiation of lipid peroxidation by citrate-Fe2+.

Animals↗
PubMed1987

Natural course of iron delocalization and lipid peroxidation during the first eight hours following a 15-minute cardiac arrest in dogs.

Lipid peroxidation is thought to be a major contributing factor in neurological injury following cardiac arrest. Because iron availability is a prerequisite for lipid peroxidation, this experiment was designed to examine the natural time course of iron release, lipid peroxidation, and cerebral polyunsaturated fatty acid content following a 15-minute cardiac arrest in dogs. Large mongrel dogs were anesthetized with ketamine and halothane and divided into three groups of five each. In two groups, cardiac arrest was induced with KCl. After 15 minutes of cardiac arrest, the dogs were resuscitated by five minutes of internal cardiac massage, epinephrine, bicarbonate, and internal defibrillation. All ten dogs were resuscitated and supported by a standard intensive care protocol until tissue harvest. A 3-g portion of parietal cerebral cortex was obtained from the nonischemic dogs (n = 5), or at two hours (n = 5), or eight hours (n = 5) after resuscitation. Total tissue iron was measured by an atomic emission spectrometer; low molecular weight species (LMWS) iron by the o-phenanthroline test on an ultrafiltered sample; and lipid peroxidation by both the thiobarbituric acid test (TBARS) and determination of the tissue content of lipid double bonds, calculated by first fractionating the lipids by gas-liquid chromatography and then measuring the double bonds in each fraction by spectrometry and summing the results. Univariate ANOVA demonstrated all variables except total tissue iron to have significance at P less than .02. At two hours of reperfusion, LMWS iron and TBARS were significantly elevated above nonischemic control levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Short-term oral administration of polybrominated biphenyls enhances the development of hepatic enzyme-altered foci in initiated rats.

FireMaster BP-6 (FM), a commercial mixture of polybrominated biphenyls (PBB), has been shown to act as a tumor promoter in hepatocarcinogenesis assays in rats. Most hepatic tumor promoters must be administered for many weeks or months. Because FM is highly persistent in animal tissues, it was hypothesized that very short-term administration of FM would result in tumor promotion. Female Sprague-Dawley rats weighing 185-215 g were initiated by a two-thirds partial hepatectomy followed by 10 mg diethylnitrosamine/kg body weight (BW) 24 h later. Thirty days later, rats were gavaged with FM in corn oil, at total doses of 0, 13, or 130 mg FM/kg BW. Half the dose was given on d 30, and the remaining half was given 24 h later. At 120 d after gavage the rats were killed and necropsied. Five liver sections from each animal were histochemically stained for gamma-glutamyl transpeptidase-positive enzyme-altered foci (EAF). EAF were significantly increased over control values in initiated rats given 130 mg FM/kg. In animals given 13 mg FM/kg, EAF were increased to a lesser extent but not significantly above controls. Enhancement of these EAF in initiated rats reflects tumor-promoting activity. In this study, 24-h administration of FM in initiated rats was sufficient to enhance hepatic EAF measured 120 d later in an rats was sufficient to enhance hepatic EAF measured 120 d later in an initiation-promotion protocol, and a dose of 13 mg FM/kg was apparently close to a possible no-effect threshold level for enhancement of EAF.

Administration, Oral↗
PubMed1987

Biodegradation of DDT [1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane] by the white rot fungus Phanerochaete chrysosporium.

Extensive biodegradation of 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane (DDT) by the white rot fungus Phanerochaete chrysosporium was demonstrated by disappearance and mineralization of [14C]DDT in nutrient nitrogen-deficient cultures. Mass balance studies demonstrated the formation of polar and water-soluble metabolites during degradation. Hexane-extractable metabolites identified by gas chromatography-mass spectrometry included 1,1,-dichloro-2,2-bis(4-chlorophenyl)ethane (DDD), 2,2,2-trichloro-1,1-bis(4-chlorophenyl)ethanol (dicofol), 2,2-dichloro-1,1-bis(4-chlorophenyl)ethanol (FW-152), and 4,4'-dichlorobenzophenone (DBP). DDD was the first metabolite observed; it appeared after 3 days of incubation and disappeared from culture upon continued incubation. This, as well as the fact that [14C]dicofol was mineralized, demonstrates that intermediates formed during DDT degradation are also metabolized. These results demonstrate that the pathway for DDT degradation in P. chrysosporium is clearly different from the major pathway proposed for microbial or environmental degradation of DDT. Like P. chrysosporium ME-446 and BKM-F-1767, the white rot fungi Pleurotus ostreatus, Phellinus weirii, and Polyporus versicolor also mineralized DDT.

Basidiomycota↗
PubMed1986

Reductive release of iron from ferritin by cation free radicals of paraquat and other bipyridyls.

NADPH-cytochrome P-450 reductase-catalyzed reduction of paraquat promoted the release of iron from ferritin. Aerobically, iron release was inhibited approximately 60% by superoxide dismutase, whereas xanthine oxidase-dependent iron release was inhibited nearly 100%. This suggests that both superoxide and the paraquat cation radical can catalyze the release of iron from ferritin. Accordingly, under anaerobic conditions, the paraquat radical mediated a very rapid, complete release of iron from ferritin. Similarly, the cation free radicals of the closely related chemicals, diquat and benzyl viologen, also promoted iron release. ESR studies demonstrated that electron transfer from the paraquat cation radical to ferritin accounts for the reductive release of iron. The ferritin structure was not altered by exposure to the paraquat radical and also retained its ability to re-incorporate iron. These studies indicate that release of iron from ferritin may be a common feature contributing to free radical-mediated toxicities.

Animals↗
PubMed1986

Release of iron from ferritin by cardiotoxic anthracycline antibiotics.

The use of the extremely effective anthracycline antitumor drugs adriamycin and daunomycin is limited by a severe, dose-dependent cardiomyopathy. Anthracycline-induced toxicity has been proposed to involve iron-dependent oxidative damage to biological macromolecules yet little is known regarding the availability of physiologic iron. We now report that, in the presence of NADPH-cytochrome P-450 reductase, these drugs undergo redox cycling to generate superoxide which mediates a slow, reductive release of iron from ferritin, the major intracellular iron storage protein. Anaerobically, the semiquinone free radical forms of adriamycin and daunomycin catalyze a very rapid, extensive release of iron from ferritin. In contrast, diaziquone, an aziridinyl quinone antitumorigenic agent which is less cardiotoxic, is unable to release iron from ferritin. Thus, the present studies suggest that the cardiomyopathy observed with the anthracyclines, and perhaps their antineoplastic activity as well, may be related to their ability to delocalize tissue iron, thereby contributing to the formation of strong oxidants capable of damaging critical cellular constituents.

Antibiotics, Antineoplastic↗
PubMed1986

The effects of nonadecafluoro-n-decanoic acid on serum retinol and hepatic retinyl palmitate hydrolase activity in male Sprague-Dawley rats.

The effects of nonadecafluoro-n-decanoic acid (NDFDA) on serum retinol levels and hepatic retinyl palmitate hydrolase (RPH) activity were investigated in male Sprague-Dawley rats given a single intraperitoneal (IP) dose of 0, 50, or 100 mg/kg NDFDA and sacrificed at two, eight, or 11 days. Treated animals exhibited depressed serum retinol levels, lymphoid involution, and failure to gain weight in proportion to the dose. Hepatic RPH activities were depressed in both treatment groups at all time points and correlated with serum retinol levels. Hepatic retinol levels were also depressed by Day 11. Extraction of hepatic homogenates with acetone removed NDFDA and increased RPH activities twofold and threefold for the low- and high-dose groups, respectively. Analysis of partially purified RPH showed both NDFDA and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to be noncompetitive inhibitors: KI = 450 and 750 microM, respectively. We conclude that NDFDA causes a decrease in the mobilization of vitamin A from the liver by noncompetitive inhibition of RPH.

Animals↗