Biomedical subjects

M L Hu

Publications and source records attributed to M L Hu.

At least 37 records · Page 2Linked to original sources

PubMed1994

Interactions of human blood plasma with hydrogen peroxide and hypochlorous acid.

Activated neutrophils produce both hydrogen peroxide (H2O2) and hypochlorous acid (HOCl). Previous work has shown that HOCl depletes antioxidants, modifies proteins, and forms fatty acid chlorohydrins but does not cause significant lipid peroxidation in human plasma. Because activated phagocytes have been claimed to stimulate lipid peroxidation in plasma, we examined the effects of H2O2 and HOCl alone and in combination on plasma constituents. Hydrogen peroxide at concentrations below 0.5 mmol/L had little effect, but 1 to 2 mmol/L H2O2 caused loss of ascorbic acid and protein thiol groups, an effect potentiated by preincubation of the plasma with sodium azide to inhibit catalase. H2O2 caused no detectable lipid peroxidation or loss of alpha-tocopherol in plasma, but some depletion of ubiquinol occurred. The combination of HOCl and H2O2 caused more lipid peroxidation than either agent alone. Peroxidation was not inhibited by the metal chelators ethylenediaminetetraacetic acid and deferoxamine or by the singlet O2/hydroxyl radical scavenger histidine. We hypothesize that the phagocyte-derived H2O2 and HOCl could interact in the microenvironment of the activated leukocyte to induce lipid peroxidation of plasma lipoproteins or cell membranes (or both).

Antioxidants↗

PubMed1993

Cigarette smoke oxidation of human plasma constituents.

In vitro exposure of fresh human plasma to cigarette smoke (CS) was used as a model for reactions that could be occurring in CS-exposed respiratory tract lining fluids (RTLFs) and lung parenchyma. The central focus of this model was to characterize the consumption of endogenous plasma antioxidants in relationship to the appearance of oxidized proteins and lipids as a consequence of exposure to CS, or to aldehydes present in CS. The amelioration of CS-induced protein and lipid oxidation in plasma by the addition of selective exogenous antioxidants was also assessed. We found that: (i) exposure of human plasma to gas phase CS causes both lipid peroxidation and protein oxidation, and endogenous ascorbic acid protects against lipid, but not protein, oxidation; (ii) whole CS causes protein oxidation, but does not induce lipid peroxidation; (iii) addition to plasma of aldehydes known to be present in CS causes protein damage, but does not induce either lipid peroxidation or oxidation of ascorbic acid; and (iv) exogenously added dihydrolipoic acid (DHLA) preserves ascorbic acid levels in plasma exposed to the gas phase of CS, and protects, to some extent, against lipid peroxidation; DHLA also protects against protein oxidation, whereas added glutathione (GSH) only protects against protein, but not lipid, oxidation.

Adult↗

PubMed1993

No detrimental effects in delaying initiation of gonadotropin administration after pituitary desensitization with gonadotropin-releasing hormone agonist.

OBJECTIVE: To determine if delaying initiation of exogenous gonadotropin administration after pituitary desensitization with gonadotropin-releasing hormone agonist (GnRH-a) is a realistic option to avoid scheduling clinical and laboratory work on weekends/holidays. DESIGN, PATIENTS: A review of 57 in vitro fertilization (IVF) cycles in which, after pituitary desensitization with GnRH-a, initiation of gonadotropin administration were delayed in an attempt to avoid off-hour work. Thirty-eight IVF cohort cycles served as control. SETTING: Tertiary medical center. RESULTS: There were no statistically significant differences in ovarian response, dose of gonadotropin required, oocytes and embryos obtained, pregnancy rates, and abortion rates between groups. Eighty-three percent of the delayed cycles had clinical and laboratory work that fell within weekdays. CONCLUSION: Delaying initiation of exogenous gonadotropin administration after pituitary desensitization had no detrimental effects on IVF outcomes. It may be used to avoid scheduling work on weekends/holidays.

Adult↗

PubMed1993

Oxidation of biologic molecules by ozone: the effect of pH.

Ozone (O3) is a powerfully oxidizing pollutant gas. Its toxic effects to animals appear to be worsened by coexposures to acid-generating compounds such as oxides of nitrogen and sulfur. Ozone (16 ppm) oxidizes ascorbic acid and uric acid (two important antioxidants in lung lining fluids) at equal rates at pH 5.0 or pH 7.4. Loss of intrinsic fluorescence and formation of carbonyls in albumin exposed to O3 are similar at both pH values. However, albumin-SH groups are lost much faster on exposure to O3 at pH 7.4 than at acidic pH values. A similar slower rate of -SH group disappearance at acidic pH is seen when cysteine or reduced glutathione are exposed to O3. We suggest that the ability of reduced glutathione, albumin, and other proteins containing -SH groups to scavenge O3 in the respiratory tract is impaired at low pH and that this effect could contribute to the aggravation of O3 toxicity.

Albumins↗

PubMed1993

Antioxidant protection against hypochlorous acid in human plasma.

Hypochlorous acid (HOCI) is a powerful oxidizing and chlorinating agent produced by the neutrophil enzyme myeloperoxidase. The antioxidant defenses of freshly prepared human plasma against HOCI/OCI- were explored. Addition of HOCI/OCI- to plasma caused rapid oxidation of ascorbic acid and thiol (-SH) groups but not of uric acid. Plasma -SH groups (which are known to be largely located on albumin) were quantitatively the most important scavenger of HOCI/OCI-, but adding extra ascorbate to plasma caused this molecule to have a more important scavenging role against HOCI/OCI-. Added HOCI/OCI- produced no detectable lipid peroxidation in plasma or depletion of lipid-soluble antioxidants (alpha-tocopherol or ubiquinol-10). No evidence of oxidative damage to protein amino acid residues (other than -SH) was detected by the carbonyl assay. It seems that -SH groups are a major target of attack by HOCI/OCI- in vivo, and plasma albumin may be an important protective antioxidant. Ascorbic acid might also play a protective role, especially in individuals supplemented with this vitamin. Ascorbate might also be important in extracellular fluids with low albumin concentrations such as synovial, respiratory tract lining, and cerebrospinal fluids.

Antioxidants↗

PubMed1992

Interaction of nitrogen dioxide with human plasma. Antioxidant depletion and oxidative damage.

Nitrogen dioxide (NO2.) is often present in inhaled air and may be generated in vivo from nitric oxide. Exposure of human blood plasma to NO2. caused rapid losses of ascorbic acid, uric acid and protein thiol groups, as well as lipid peroxidation and depletions of alpha-tocopherol, bilirubin and ubiquinol-10. No increase in protein carbonyls was detected. Supplementation of plasma with ascorbate decreased the rates of lipid peroxidation, alpha-tocopherol depletion and loss of uric acid. Uric acid supplementation decreased rates of lipid peroxidation but not the loss of alpha-tocopherol. We conclude that ascorbic acid, protein -SH groups, uric acid and alpha-tocopherol may be important agents protecting against NO2. in vivo. If these antioxidants are depleted, peroxidation of lipids occurs and might contribute to the toxicity of NO2..

Adult↗

PubMed1992

Modification of plasma proteins by cigarette smoke as measured by protein carbonyl formation.

Exposure of human plasma to gas-phase (but not to whole) cigarette smoke (CS) produces oxidative damage to lipids [Frei, Forte, Ames & Cross (1991) Biochem. J. 277, 133-138], which is prevented by ascorbic acid. The ability of CS to induce protein damage was measured by the carbonyl assay and by loss of enzyme activity and protein -SH groups. Both whole and gas-phase CS caused formation of carbonyls in human plasma, which was partially inhibited by GSH but not by ascorbic acid or metal-ion-chelating agents. Isolated albumin exposed to CS showed much faster carbonyl formation (per unit protein) than did whole plasma; damage to isolated albumin was partially prevented by chelating agents. Isolated creatine kinase (CK) lost activity upon exposure to CS much faster than did CK in plasma. Direct addition to plasma of mixtures of some or all of the aldehydes reported to be present in CS caused protein carbonyl formation and inactivation of CK, but neither occurred to the extent produced by CS exposure.

Adult↗

PubMed1992

Glutathione and antioxidants protect microsomes against lipid peroxidation and enzyme inactivation.

The study investigated the relationship between lipid peroxidation and enzyme inactivation in rat hepatic microsomes and whether prior inactivation of aldehyde dehydrogenase (ALDH) exacerbated inactivation of other enzymes. In microsomes incubated with 2.5 microM iron as ferric sulfate and 50 microM ascorbate, ALDH, glucose-6-phosphatase (G6Pase) and cytochrome P450 (Cyt-P450) levels decreased rapidly and concurrently with increased levels of thiobarbituric acid-reactive substances. Microsomal glutathione S-transferase and nicotinamide adenine dinucleotide phosphate-cytochrome c reductase were little affected during 1 hr of incubation. Addition of reduced glutathione partially protected and N,N'-diphenyl-p-phenylenediamine and butylated hydroxytoluene completely protected microsomes against inactivation of ALDH, G6Pase and Cyt-P450, as well as lipid peroxidation induced by iron and ascorbate. ALDH was more susceptible than G6Pase to inactivation by iron and ascorbate, and was thus an excellent marker for oxidative stress. Inhibition of ALDH by cyanamide injection of rats exacerbated the inactivation of G6Pase in microsomes incubated with 0.1 mM, but not 25 microM 4-hydroxynonenal (4-HN). 4-HN did not stimulate lipid peroxidation. Thus, 4-HN may play a minor role in microsomal enzyme inactivation. In contrast, lipid peroxyl radicals play an important role in microsomal enzyme inactivation, as evidenced by the prevention of both lipid peroxidation and enzyme inactivation by chain-breaking antioxidants.

Aldehyde Dehydrogenase↗

PubMed1992

Potentiation of oxidative damage to proteins by ultraviolet-A and protection by antioxidants.

We have studied the damage of alcohol dehydrogenase (ADH) and glyceraldehyde 3-phosphate dehydrogenase (GAPD) induced by Fe++/EDTA + H2O2 in combination with UV-A (main output at 365 nm). Enzyme inactivation, formation of hydroxyl radicals (measured in the absence of enzymes), increase in protein carbonyls, oxidation of sulfhydryl (SH) groups, loss of native protein fluorescence, and enhanced protease degradation were used to determine protein damage. Hydroxyl radical production was greatly enhanced by the combination of UV-A with Fe++/EDTA + H2O2. The combined treatment increased protein carbonyls but decreased native protein fluorescence and SH groups. The combined treatment caused turbidity in GAPD but not in ADH, whereas trypsin susceptibility was increased more in ADH than in GAPD. These measurements of protein oxidation correlated well with enzyme activities. Glyceraldehyde 3-phosphate dehydrogenase and dithiothreitol were most protective against such damage, while hydroxyl radical and singlet oxygen scavengers were partially effective. Superoxide dismutase had no effect. Thus, UV-A potentiation of protein damage induced by FE++/EDTA + H2O2 appeared to involve hydroxyl radicals and perhaps singlet oxygen but not superoxide radicals. The damage to proteins induced by combination of UV-A with physiological oxidants, iron ions and H2O2 may be relevant to UV-A-induced skin and tissue damage.

Alcohol Dehydrogenase↗

PubMed1991

Vitamin E, diethylmaleate and bromotrichloromethane interactions in oxidative damage in vivo.

In vivo interactions of vitamin E with diethylmaleate (DEM) and bromotrichloromethane (CBrCl3) were examined in rats fed a diet either without vitamin E or supplemented with 30 IU dl-alpha-tocopheryl acetate/kg. Groups of rats within each dietary group were given two injections 30 min apart. One group received two injections of the mineral oil carrier. The other groups were injected with either DEM and mineral oil, mineral oil and CBrCl3, or DEM and CBrCl3. The rats were killed 10 min after the second injection. Measurements were made of hepatic GSH, thiobarbituric acid-reactive substances (TBARS) as a lipid peroxidation index, and 11 enzymes as potential markers of oxidant damage. Special focus was placed on reactive cysteine-containing aldehyde dehydrogenase (ALDH). Although dietary vitamin E protected ALDH, the enzyme was highly susceptible to oxidant damage. ALDH activity was correlated with GSH (r = 0.83, p less than 0.001) and there was an inverse relationship between the logarithmic values of ALDH activity and TBARS (r = 0.78, p less than 0.001). Similar results were observed for a number of other enzymes when GSH depletion preceded oxidant treatment. Two-way analysis of variance revealed significant effects of vitamin E and of injection treatments on hepatic GSH. There was a significant interaction between vitamin E and the injection treatments on the activities of five enzymes. The results suggested that vitamin E and GSH functioned together to protect sensitive enzymes against oxidant stress. The sensitive enzymes may be useful markers of hepatic damage in vivo.

Aldehyde Dehydrogenase↗

PubMed1990

Effect of dietary menhaden oil and vitamin E on in vivo lipid peroxidation induced by iron.

Weanling rats were fed diets containing 10% menhaden oil (MO) or 10% corn oil-lard (1:1, COL) with low (less than or equal to 5 IU/kg) or supplementary (35 IU/kg) vitamin E for six weeks. The rats were killed 30 min after injection with 24 mg iron/kg as ferrous chloride because thiobarbituric acid-reactive substances (TBARS) in liver homogenates were highest at 30 min after injection of iron into rats fed a standard diet. Tissue homogenates were used either without incubation (zero-time) or after incubation at 37 degrees C for 1 hr. In addition to TBARS and conjugated dienes, headspace hexanal and total volatiles (TOV) determined by capillary gas chromatography were useful indices of lipid peroxidation since they were decreased by vitamin E supplementation and were increased with increasing iron dose. Regardless of the dietary lipid used, vitamin E supplementation decreased headspace hexanal, TOV, TBARS and conjugated dienes in both zero-time and incubated homogenates of liver and kidney. Dietary MO increased TBARS in both zero-time and incubated homogenates of tissue from rats injected with iron. In contrast, dietary MO decreased hexanal and TOV in incubated tissue homogenates. The study demonstrated the usefulness and limitations of using hexanal and TOV as indices of lipid peroxidation.

Animals↗

PubMed1990

Lipid peroxidation in rat tissue slices: effect of dietary vitamin E, corn oil-lard and menhaden oil.

Rats were fed for 5 weeks either 10% (w/w) menhaden oil (MO) or a 10% corn oil-lard (COL) mixture (1:1) in diets with less than or equal to 5 IU or less than or equal to 2 IU/kg vitamin E, respectively, or the same diets supplemented with d-alpha-tocopheryl succinate to a total of 35 and 180 IU vitamin E/kg, respectively. Slices of liver and heart from these rats were used to study lipid peroxidation in vitro. Thiobarbituric acid-reactive substances (TBARS) were measured in the medium after incubation of the slices at 37 degrees C for 1 hr in the absence (uninduced) and presence of 0.5 mM tert-butyl hydroperoxide (induced). The release of TBARS from slices of heart and liver from rats fed either lipid decreased with increasing levels of dietary vitamin E. At the same level of dietary vitamin E, TBARS release was greater for slices of liver and heart from the MO-fed rats than from the COL-fed rats. Application of the TBARS data to a model simulating the experimental conditions showed a good correlation (r = 0.95, p less than 0.001) between experimental and simulated values. Of the 16:0-22:6 fatty acids measured in liver from MO-fed rats, 15.4% was n-6 fatty acids and 29.9% was n-3 fatty acids; in liver from COL-fed rats, the respective values were 37.4% and 3.7%. Liver and kidney vitamin E levels were unaffected by the dietary lipid.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1990

Oxidant stress inhibits the endogenous production of lipoxygenase metabolites in rat lungs and fish gills.

Hydroperoxides are potent initiators of lipid peroxidation in vivo. Acyl hydroperoxides may also regulate various aspects of lipid metabolism. In this study we investigated the regulation of the endogenous 12 lipoxygenase in trout gill and rat lung, a prominent acyl hydroperoxide catalyst in these tissues. Initial experiments revealed that the enzyme from trout gill was activated by hydroperoxides at low levels and inactivated by the same hydroperoxides at high levels. Homogenization of these tissues resulted in the production of a predominant metabolite class from released endogenous polyunsaturated fatty acids, the 12 lipoxygenase products. In rat lung, arachidonic acid was the major polyunsaturated fatty acid released and 12 (S) HETE was the major metabolite. In trout gill 20:4, 20:5 n3, and 22:6 n3 were released and the 12(S), 12, and 14 hydroxy derivatives the corresponding metabolites. Computer simulations of the sensitivity of these enzymes to hydroperoxides predicted that exogenous oxidant stress would reduce significantly the production of HETEs. Tertiary butyl hydroperoxide was added to tissue homogenates and resulted in elimination of greater than 95% of the lipoxygenase activity. These results suggest that the lipoxygenase enzyme in lung and gill tissue is a major potential source for acyl hydroperoxides in vivo, but is also very sensitive to oxidant stresses including the acyl hydroperoxides themselves. This enzyme could thus be an important focus for oxidant injury in lungs.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗

PubMed1990

Dietary supplements of vitamin E, beta-carotene, coenzyme Q10 and selenium protect tissues against lipid peroxidation in rat tissue slices.

A tissue slice model was employed to assess the effects of dietary antioxidant supplements on lipid peroxidation. In one experiment, rats were fed diets containing, either alone or in combination, vitamin E, selenium, beta-carotene or coenzyme Q10 for 42 d, and the extent of spontaneous and induced lipid peroxidation was determined by release of thiobarbituric acid-reactive substances (TBARS) into the medium. Vitamin E exhibited the greatest protection against lipid peroxidation in liver, heart and spleen; in kidney, selenium was most protective. Coenzyme Q10 was active against lipid peroxidation induced by tertbutyl hydroperoxide (t-BHP). In a second experiment, rats were fed diets containing varying amounts of vitamin E, selenium, beta-carotene and coenzyme Q10 for 30 d. Spontaneous lipid peroxidation in liver, kidney and heart decreased with increasing levels of dietary antioxidants. With increasing amounts of antioxidants, there was a diminution in TBARS released by liver and kidney slices incubated with t-BHP; in heart, only the highest levels of antioxidants significantly decreased production of TBARS. Inverse correlations between dietary vitamin E and TBARS, tissue vitamin E and TBARS, and tissue selenium-glutathione peroxidase and TBARS were highly significant. The procedure used here can evaluate dietary supplements that may find practical applications in decreasing the oxidant radical portion of disease processes.

Animals↗

PubMed1989

Rapid headspace gas chromatography of hexanal as a measure of lipid peroxidation in biological samples.

A rapid, sensitive and convenient capillary gas chromatographic-headspace method was developed to determine hexanal as an important volatile decomposition product of hydroperoxides formed from n-6 polyunsaturated fatty acids in rat liver samples. Total volatiles were also determined as a measure of overall lipid peroxidation. Samples of headspace taken from sealed serum bottles incubated at 37 degrees C were injected into a gas chromatograph. It was possible to make 15 determinations per hour. This method is convenient because no special sample manipulations are necessary. The addition of 0.5 mM ascorbic acid prior to gas chromatographic analysis significantly increased hexanal production. The applicability of the method was demonstrated in studies of the effect of iron in the presence or absence of hydroperoxides of methyl linoleate and methyl linolenate and tert-butyl hydroperoxide on rat liver homogenates, slices and microsomes. A rapid silica cartridge chromatographic procedure was used to purify hydroperoxides from autoxidized methyl linoleate and methyl linolenate, and hydroperoxy epidioxides (cyclic peroxides) from autoxidized methyl linolenate in 20-40 mg quantities. The hydroperoxides and hydroperoxy epidioxides of methyl linolenate were effective inducers of n-6 polyunsaturated fatty acid peroxidation in liver homogenates. Hexanal and thiobarbituric acid-reacting substances were significantly correlated in liver homogenates and microsomes but not in slices. This specific method for hexanal, a known product of peroxidation of n-6 polyunsaturated fatty acids, can be used as a good measure of lipid peroxidation.

Aldehydes↗

PubMed1989

Effect of dietary lipids and vitamin E on in vitro lipid peroxidation in rat liver and kidney homogenates.

Rats were fed for 5 wk 10% (wt/wt) menhaden oil (MO) or a 10% corn oil-lard (COL) mixture (1:1) in diets with a low vitamin E content (less than or equal to 5 mg/kg) or supplemented with d-alpha-tocopheryl succinate to a total of 30 or 150 mg per kg. Thiobarbituric acid-reactive substances (TBARS), conjugated dienes (CD), hexanal and total volatiles (TOV) were measured in tissue homogenates incubated at 37 degrees C for 1 h in the absence (uninduced) and presence of 15 microM ferrous sulfate (induced). The fatty acid composition of liver and kidney reflected that of dietary lipids. For uninduced peroxidation, there was in general a significant inverse correlation of TBARS, CD and TOV with the log of dietary vitamin E content for liver and kidney from rats fed either lipid. For induced peroxidation, the inverse correlation was significant for liver, but not for kidney, from rats fed either lipid. The correlation was generally higher for liver and kidney from rats fed COL than for tissues from rats fed MO. Vitamin E was thus a more effective antioxidant for liver than for kidney regardless of the dietary lipid, and for liver and kidney from rats fed COL than from rats fed MO. Dietary MO enhanced tissue susceptibility to both peroxidation systems. A simulation model developed to mimic the experiments showed good correlations between experimental data and simulated values.

Analysis of Variance↗

PubMed1988

In vivo effects of aurothioglucose and sodium thioglucose on rat tissue sulfhydryl levels and plasma sulfhydryl reactivity.

The active component of aurothioglucose (ATG) in effecting changes in plasma sulfhydryl (SH) levels and plasma SH reactivity with 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) was determined. These two measurements are applied clinically to rheumatoid arthritis patients undergoing chrysotherapy. Normal rats were injected intramuscularly daily for seven days with 30 mumol of either ATG or sodium thioglucose (STG)/kg body wt or with an equivalent volume of the carrier, 0.05% benzyl alcohol. ATG but not STG significantly increased total SH levels in plasma, liver, and kidney. The seven-day treatment with ATG significantly increased glutathione levels in kidney but not in liver or plasma. Thus, gold(I) rather than thioglucose was the active moiety that affected SH levels in ATG-injected rats. In vivo, gold(I) was also the active moiety that stimulated plasma SH reactions with DTNB at pH 7.4, since injection of ATG but not STG stimulated the SH reactivity in fresh plasma. In vitro, ATG increased the rate of plasma reaction with DTNB at pH 7.4, thus, gold(I) ions acted as a catalyst in the SH-disulfide exchange reaction. This study demonstrates that gold(I) but not its thiol ligand strongly interacts with protein SH groups in the rat tissues. Such an interaction may play an important role in the biological actions of gold.

Albumins↗

PubMed1988

Effect of chronic aurothioglucose treatment of rats on kidney catalase and cytochromes.

Catalase activity and cytochrome content were measured in kidneys of Fisher 344 rats injected with aurothioglucose (ATG) either daily for 3 days or 5 days a week for up to 8 wk. Catalase activity was decreased 39%, 59%, and 48% (all p less than 0.001) after 3 days, 2 wk, and 8 wk, respectively. Microsomal cytochrome P-450 levels decreased 71%, 86%, and 80% (all p less than 0.001) after 3 days, 2 wk, and 8 wk, respectively. In contrast, cytochrome b5 was significantly increased at 3 days and 2 wk, but not at 8 wk. Microsomal heme contents decreased 44% (p less than 0.001), 34% (p less than 0.001), and 22% (p greater than 0.05) at 3 days, 2 wk, and 8 wk, respectively. The content of mitochondrial cytochromes aa3, b, c1, and c were not affected after 8 wk of ATG treatment. In vitro inhibition of the heme-containing enzyme delta-aminolevulinic acid dehydratase by ATG was reversible in the presence of physiological concentrations of small thiols. Although the activity of this enzyme in kidneys of ATG-treated rats was not measured, its significant inhibition in vivo by ATG appears unlikely. This study demonstrates that there were differential effects of gold on the various cytochromes and that changes in catalase activity paralleled changes in cytochrome P-450 and heme contents in the kidneys of ATG-treated rats. The findings are relevant to nephrotoxicity during chrysotherapy.

Animals↗