Exercise performance, red blood cell deformability, and lipid peroxidation: effects of fish oil and vitamin E

Oostenbrug, G. S., R. P. Mensink, M. R. Hardeman, T. DeVries, F. Brouns, and G. Hornstra. Exercise performance, red bloodcell deformability, and lipid peroxidation: effects of fish oil andvitamin E. J. Appl. Physiol. 83(3):746-752, 1997.Previous studies have indicated that fish oilsupplementation increases red blood cell (RBC) deformability, which mayimprove exercise performance. Exercise alone, or in combination with anincrease in fatty acid unsaturation, however, may enhance lipidperoxidation. Effects of a bicycle time trial of ~1 h on RBCcharacteristics and lipid peroxidation were, therefore, studied in 24 trained cyclists. After 3 wk of fish oil supplementation (6 g/day),without or with vitamin E (300 IU/day), trial performance,RBC characteristics, and lipid peroxidation were measuredagain. RBC deformability appeared to decrease duringendurance exercise. After correction for hemoconcentration, plasmatotal tocopherol concentrations decreased by 0.77 µmol/l(P = 0.012) or 2.9% and carotenoidconcentrations by 0.08 µmol/l (P = 0.0008) or 4.5%. Endurance exercise did not affect the lag time andrate of in vitro oxidation of low-density lipoproteins (LDLs), but themaximum amount of conjugated dienes formed decreased by 2.1 ± 1.0 µmol/mmol LDL cholesterol (P = 0.042) or 1.2%. Fish oil supplementation with andwithout vitamin E did not affect RBC characteristics or exerciseperformance. Both supplements decreased the rate of LDL oxidation, andfish oil supplementation with vitamin E delayed oxidation. The amountof dienes, however, was not affected. The supplements also did notchange effects of exercise. We conclude that the changes observedduring endurance exercise may indicate increased oxidative stress, butfurther research is necessary to confirm this. Fish oil supplementation does not improve endurance performance, but it also does not cause oraugment changes in antioxidant levels or LDL oxidation during exercise.

     

Effects of antioxidant vitamin supplements on Helicobacter pylori-induced gastritis in Mongolian gerbils

BACKGROUND: Epidemiological studies show that high intake of food-bound vitamin C and E reduces the risk of gastric cancer. Whether dietary supplementation with antioxidant micronutrients interferes with Helicobacter pylori infection and associated diseases is unclear. The aim of this study was to investigate if dietary vitamin C or E supplementation influences the progression of gastritis, gastric mucosal nitrosative and oxidative protein damage, gastric mucosal lipid peroxidation, or gastric mucosal oxidative DNA damage in H. pylori-infected Mongolian gerbils. MATERIALS AND METHODS: Gerbils were divided into four groups: H. pylori-infected animals fed with vitamin C- or vitamin E-supplemented food, and infected and uninfected animals given standard rodent food. Subgroups of animals were killed at different time-points until 52 weeks postinfection. Concentrations of 3-nitrotyrosine and thiobarbituric acid-reactive substances (TBARS) in the gastric mucosa were determined with an immunodot blot and a fluorometric method, respectively. Mucosal concentrations of carbonyl carbons on proteins and 8-hydroxydeoxyguanosine were determined by enzyme-linked immunosorbent assay. Gastritis was scored semiquantitatively. RESULTS: Vitamin supplements had no effect on the colonization with H. pylori. Vitamin C as well as vitamin E supplements reduced mucosal 3-nitrotyrosine concentrations to normal levels in infected animals. Vitamin E supplements decreased mucosal protein carbonyls and TBARS in short-term gastritis. In addition, vitamin C supplements caused attenuated mucosal oxidative DNA damage and milder mucosal inflammation in short-term gastritis. CONCLUSION: Vitamin C or vitamin E supplementation leads to some short-term protective effects on H. pylori-induced gastritis in Mongolian gerbils. These effects seem to subside over time when the infection persists.     

Effect of vitamin E supplementation on arsenic induced alteration in blood biochemical profile,oxidant/antioxidant status,serum cortisol level and retention of arsenic and selenium in goats

Arsenic (As) exerts oxidative stress with depletion of body selenium in monogastric animals. But in ruminants this fact is not yet verified. Vitamin E is an effective dietary antioxidant. Thus, in this experiment, the protective effect of vitamin E against arsenic toxicity induced by sodium arsenite (60 mg As/kg diet) was investigated in goat kids. For this, 21 male kids were divided into three equal groups and fed either basal diet as such (control), or supplemented with 60 mg As/kg diet and 60 mg As/kg diet + 250 IU vitamin E/kg diet for 180 days. Vitamin E supplementation alleviated the toxic effects caused by arsenic on serum alanine aminotransferase and aspartate aminotransferase and lipid peroxidation. It also prevented the depletion of reduced glutathione content and reduction in activity of catalase, superoxide dismutase and glutathione-s-transferase in erythrocytes resulted from arsenic intoxication. The elevated levels of arsenic and reduced levels of selenium in the serum and tissues in arsenic treated animals were attenuated by vitamin E supplementation, though not completely. However, serum cortisol level was not affected by arsenic. It was concluded that arsenic exerts cortisol independent stressor mechanism and supplementation of vitamin E at a level of 250 IU/kg diet was partially effective in reducing tissue accumulation of arsenic in the body and protect the kids from oxidative stress induced by arsenic.     

Effects of supplementation with vitamin E on the performance and the tissue peroxidation of broiler chicks and the stability of thigh meat against oxidative deterioration

Two experiments were conducted: Expt 1 determined the optimal allowance of vitamin E in the diet for broiler chicks aged 0–3 weeks; Expt 2 investigated the effects of different dietary levels of vitamin E (α-tocopherol) on the performance and the oxidative stability of thigh meat of broiler chicks during storage. In Expt 1, 1-day-old 900 broiler chicks were allocated to five treatments, each with six replicates (cages) of 22 as-hatched chicks for performance evaluation, and another cage of 45 male chicks for determining plasma and hepatic α-tocopherol and thiobarbituric acid reactive substances (TBARS) concentration in blood and liver. The basal dietary α-tocopherol concentration was 13 mg/kg, and the five α-tocopherol acetate supplementation levels were 0, 5, 10, 50 and 100 mg/kg. For 0–3-week-old broiler chicks fed with maize–soya bean meal–soya oil type diet, supplementation of vitamin E did not influence the feed intake, but tended to improve growth and feed utilization, however there was no significant correlation between performance and vitamin E supplementation level. Significant positive correlations existed between dietary supplemental vitamin E level and plasma or hepatic α-tocopherol concentrations (P<0.05), and a negative correlation with hepatic TBARS levels no matter at what age (11, 16 and 21 days). In Expt 2, 2200 broiler chicks were randomly allocated to five treatments with four replicates (pens) in each. Chicks were fed ad libitum five pellet diets supplemented with vitamin E at 5, 10, 20, 50 and 100 mg/kg of diet, respectively. The basal dietary α-tocopherol level of grower and finisher diets were 7 and 6 mg/kg, respectively. Supplementation of vitamin E tended to improve growth and feed utilization of birds during 0–3 weeks of age, but the performance from 0 to 6 weeks of age were not influenced. The hepatic α-tocopherol concentrations of 6-week-old chicks linearly increased with the dietary vitamin E levels (R²=0.98, P<0.001). The content of TBARS in the thigh meat over 4 days of storage under 4°C was significantly decreased by increasing dietary vitamin E level (P<0.05). There was a significant inverse relationship between TBARS value in the thigh meat and the dietary vitamin E level (R²=0.93, P<0.01). Supplementation of vitamin E significantly improved the meat quality stability substantially against oxidative deterioration. Comparing the hepatic α-tocopherol levels of chicks in Expts 1 and 2, total allowance of dietary α-tocopherol of 20–30 mg/kg could sustain relatively constant hepatic α-tocopherol level at round about 2–2.5 μg/kg.     

Regulation of expression of antioxidant enzymes by vitamin E and curcumin in <Emphasis Type="SmallCaps">l</Emphasis>-thyroxine-induced oxidative stress in rat renal cortex

The present study investigates the antioxidative effects of vitamin E and curcumin against l-thyroxine (T₄)-induced oxidative stress in renal cortex of adult male rats. Rats were made hyperthyroid by administration of l-thyroxine (0.0012%) in their drinking water for 30 days. Vitamin E (200 mg/kg body weight/day) and curcumin (30 mg/kg body weight/day) were supplemented singly or in combination orally for 30 days along with l-thyroxine treatment. The elevated level of oxidative stress parameters (lipid peroxidation and protein carbonylation) and decline level of small antioxidant molecules (reduced glutathione and ascorbic acid) in renal cortex of T₄-treated rats were restored back by supplementation of vitamin E or/and curcumin. Increased superoxide dismutase and catalase activities in kidney cortex of T₄-treated rats were ameliorated in response to vitamin E or/and curcumin treatment. The elevated translated product of Cu/Zn-SOD, Mn-SOD and catalase in T₄-treated rats were differentially reduced by the administration of vitamin E and curcumin independently or in combination. Cu/Zn-SOD expression was ameliorated by both vitamin E and curcumin independently or in combination, whereas Mn-SOD expression was ameliorated by the supplementation of vitamin E or curcumin independently. However, the expression of catalase was alleviated by only supplementation of vitamin E to T₄-treated rats. The results suggest that both vitamin E and curcumin may play an important role in protecting T₄-induced oxidative stress in rat renal cortex by differentially modulating the activities of antioxidant enzymes and oxidative stress parameters.     

Influence of dietary vitamin E and C supplementation on vitamin E and C content and thiobarbituric acid reactive substances (TBARS) in different tissues of growing pigs

To investigate the influence and possible interactions of dietary vitamin E and C supplementation on vitamin content of both vitamins and oxidative stability of different pork tissues 40 Large White barrows from 25?kg to 106?kg were allocated to four different cereal based diets: Basal diet (B), dl-α-tocopherylacetate?+?200?mg/kg (E), crystalline ascorbic acid?+?300?mg/kg (C) or both vitamins (EC). At slaughtering samples of liver, spleen, heart, kidney, backfat outer layer, ham and M. longissimus dorsi were obtained. Growth performance of the pigs and carcass characteristics were not influenced by feeding treatments. Dietary vitamin E supplementation had a significant effect on the vitamin E and α-tocopherol concentration in all investigated tissues. Backfat outer layer, liver, spleen, kidney and heart had higher vitamin E concentrations than ham and M. longissimus dorsi. Dietary vitamin C supplementation tended towards enhanced vitamin E levels except for ham samples. Therefore, some synergistic actions without dietary vitamin E supplementation between the two vitamins could be shown. The vitamin C concentration and TBARS were increased or at least equal in all tissues due to vitamin C supplementation. Dietary α-tocopherol supplementation resulted in lower TBARS in backfat outer layer (malondialdehyde 0.35?mg/kg in B vs. 0.28?mg/kg in E), but increased in heart and ham. When both vitamins were supplemented (EC) TBARS were lower in M. longissimus dorsi and backfat outer layer, equal in heart and higher in liver and ham compared to a single vitamin C supplementation. Rancimat induction time of backfat outer layer was 0.3?h higher in C compared to B and 0.17?h higher in EC than in E. Correlations between levels of both vitamins were positive for kidney (r?=?0.169), M. longissimus dorsi (r?=?0.499) and ham (r?=?0.361) and negative for heart (r?=???0.350). In liver and spleen no interaction could be found. In backfat outer layer vitamin E was positively correlated with rancimat induction time (r?=?0.550) and negatively with TBARS (r?=???0.202), but provided no evidence that dietary vitamin E supply led to better oxidative stability.     

Phospholipid Hydroperoxides and Lipid Peroxidation in Liver and Plasma of ODS Rats Supplemented with α-Tocopherol and Ascorbic Acid

Forty-five mutant male ODs rats, unable to synthesize ascorbic acid, were fed nine diets containing 5, 50 or 250 mg of vitamin E/kg diet and 150,300 or 900 mg of vitamin C/kg diet for 21 days. The concentrations of vitamins C and E increased in liver and plasma in relation to the level of these vitamins in the diet. Vitamin C dietary supplementation increased the plasma vitamin E content at low levels of vitamin E intake, supporting the concept of an in vivo synergism between both antioxidant vitamins. Vitamin C, at the dietary levels studied, did not affect the lipid peroxidation. Vitamin E decreased liver and plasma endogenous levels of thiobarbituric acid-reactive substances and liver sensitivity to non-enzymatic lipid peroxidation. This was confirmed by a highly specific assay of lipid hydroperoxides using high performance liquid chromatography with chemiluminescence detection. The hepatic concentration of both phosphatidylcholine and phosphatidylethanolamine hydroperoxides decreased as the vitamin E content of the diet increased. The results show for the first time the capacity of vitamin E to protect against peroxidation of major phospho-lipids in vivo under basal unstressed conditions.     

Influence of dietary vitamin E and C supplementation on vitamin E and C content and thiobarbituric acid reactive substances (TBARS) in different tissues of growing pigs

To investigate the influence and possible interactions of dietary vitamin E and C supplementation on vitamin content of both vitamins and oxidative stability of different pork tissues 40 Large White barrows from 25 kg to 106 kg were allocated to four different cereal based diets: Basal diet (B), dl-alpha-tocopherylacetate + 200 mg/kg (E), crystalline ascorbic acid + 300 mg/kg (C) or both vitamins (EC). At slaughtering samples of liver, spleen, heart, kidney, backfat outer layer, ham and M. tongissimus dorsi were obtained. Growth performance of the pigs and carcass characteristics were not influenced by feeding treatments. Dietary vitamin E supplementation had a significant effect on the vitamin E and alpha-tocopherol concentration in all investigated tissues. Backfat outer layer, liver, spleen, kidney and heart had higher vitamin E concentrations than ham and M. longissimus dorsi. Dietary vitamin C supplementation tended towards enhanced vitamin E levels except for ham samples. Therefore, some synergistic actions without dietary vitamin E supplementation between the two vitamins could be shown. The vitamin C concentration and TBARS were increased or at least equal in all tissues due to vitamin C supplementation. Dietary alpha-tocopherol supplementation resulted in lower TBARS in backfat outer layer (malondialdehyde 0.35 mg/kg in B vs. 0.28 mg/kg in E), but increased in heart and ham. When both vitamins were supplemented (EC) TBARS were lower in M. longissimus dorsi and backfat outer layer, equal in heart and higher in liver and ham compared to a single vitamin C supplementation. Rancimat induction time of backfat outer layer was 0.3 h higher in C compared to B and 0.17 h higher in EC than in E. Correlations between levels of both vitamins were positive for kidney (r = 0.169), M. longissimus dorsi (r = 0.499) and ham (r = 0.361) and negative for heart (r = -0.350). In liver and spleen no interaction could be found. In backfat outer layer vitamin E was positively correlated with rancimat induction time (r = 0.550) and negatively with TBARS (r = -0.202), but provided no evidence that dietary vitamin E supply led to better oxidative stability.     

A combination of ascorbic acid and α-tocopherol or a combination of Mg and Zn are both able to reduce the adverse effects of lindane-poisoning on rat brain and liver

The purpose of this study, carried out on male Wistar rats, was to evaluate the beneficial effects of supplementation with ascorbic acid (Vit C) and α-tocopherol (Vit E) or with Mg and Zn upon lindane-induced damages in liver and brain. Under our experimental conditions, lindane poisoning (5 mg/kg body weight per day for 3 days) resulted in (1) an increased level of plasma glucose, cholesterol and triglycerides, (2) an increased activity of LDH, ALP, AST, ALT, (3) an oxidative stress in liver and brain as revealed by an increased level of lipids peroxidation (TBARS) and a decrease of glutathione-peroxidase, superoxide dismutase and catalase activities in liver and brain. In conclusion, both Vit C + E or Mg + Zn treatments display beneficial effects upon oxidative stress induced by lindane treatment in liver and brain.     

Exercise training improves lusitropy by isoproterenol in papillary muscles from aged rats

Taffet, George E., Lloyd A. Michael, and Charlotte A. Tate.Exercise training improves lusitropy by isoproterenol in papillarymuscles from aged rats. J. Appl.Physiol. 81(4): 1488-1494, 1996.Aging isassociated with a decreased cardiac responsiveness to -adrenergicstimulation. We examined the effect of endurance exercise training ofold Fischer 344 male rats on -adrenergic stimulation of the functionof isolated left ventricular papillary muscle. Three groups wereexamined: sedentary mature (SM; 12-mo old), sedentary old (SO;23-24 mo old), and exercised old (EO; 23-24 mo old) that weretreadmill trained for 4-8 wk. The isometric contractile propertieswere studied at 0.2 Hz and 0.75 mM calcium. Without -adrenergicstimulation, there were no group differences for peak tension, maximumrate of tension development(+dP/dt), or maximum rateof tension dissipation(dP/dt). The time to peak tension was longer (P < 0.05) forboth EO and SO than for SM rats. Half relaxation time(RT_1/2) was prolonged(P < 0.05) for SO compared with SMand EO (which did not differ). The three groups did not differ in the-adrenergic stimulation by isoproterenol of peak tension,dP/dt, time to peak tension, orcontraction duration. The inotropic response(+dP/dt) of SM was greater(P < 0.05) than that in SO or EOrats (which did not differ); however, the lusitropic response(RT_1/2) was lesser(P < 0.05) in SO than in SM or EO rats (which did not differ). Thus exercise training of old rats improved the lusitropic response to isoproterenol without altering theage-associated impairment in inotropic response.

     

Synaptosomes isolated from Goto-Kakizaki diabetic rat brain exhibit increased resistance to oxidative stress: role of vitamin E

Increased oxidative stress is believed to be an important factor in the development of diabetic complications. In this study, the effect of diabetes on the susceptibility of synaptosomes to oxidative stress, induced by the oxidizing system ascorbate/Fe2+, on the activity of antioxidant enzymes and on the levels of glutathione and vitamin E was investigated. Synaptosomes were isolated from brain of 29-weeks-old Goto-Kakizaki (GK) rats, a model of non-insulin dependent diabetes mellitus and from normal Wistar rats. Synaptosomes isolated from GK rats displayed a lower susceptibility to lipid peroxidation, as assessed by quantifying thiobarbituric acid reactive substances (TBARS), than normal rats (5.33 +/- 0.79 and 7.58 +/- 0.7 nmol TBARS/mg protein, respectively). In the absence of oxidants, no significant differences were found between the levels of peroxidation in synaptosomes of diabetic or control rats. Superoxide dismutase (SOD), glutathione peroxidase and glutathione reductase activities were unaltered in the brain of diabetic rats. There were no statistically significant differences in fatty acid composition of total lipids and reduced glutathione levels in synaptosomes of diabetic and control rats. The decreased susceptibility to membrane lipid peroxidation of diabetic rats synaptosomes correlated with a 1.3-fold increase in synaptosomal vitamin E levels. Vitamin E levels in plasma were also higher in diabetic rats (21.32 micromol/l) as compared to normal rats (15.13 micromol/l). We conclude that the increased resistance to lipid peroxidation in GK rat brain synaptosomes may be due to the increased vitamin E content, suggesting that diabetic animals might develop enhanced defense systems against brain oxidative stress.     

Comparative study of oxidative stress and antioxidative markers in patients infected with Toxoplasma gondii

Recently, oxidative stress and antioxidative compounds have been described as potential biomarkers. However, there is no consensus on the most appropriate oxidative and antioxidative biomarkers for patients with Toxoplasma gondii. In the present study, we evaluated the levels of lipid, protein, DNA oxidative damage and antioxidants in samples from patients infected with T. gondii with and without ocular toxoplasmosis. The levels of MDA, TBARS, micronuclei, carbonyl, GSH, vitamin C and vitamin E were measured on samples from 8 patients positive for T. gondii antibodies with ocular toxoplasmosis (OT), 20 patients positive for T. gondii antibodies without ocular toxoplasmosis (non OT), and 12 healthy individuals negative for T. gondii antibodies. The levels of MDA, TBARS, carbonyl and micronuclei were significantly higher in non OT patients, while MDA and TBARS levels were lower in OT patients. In contrast, the antioxidative factors, GSH and vitamin E levels were significantly lower in non OT patients, while vitamin C was lower in non OT and OT patients. Additionally, non OT patients were indicated to be high producers of oxidative markers (TBARS, MDA, micronuclei and carbonyl), while control group was indicated to be high producer of antioxidative markers (GSH, vitamins C and E). However, OT patients were not found as high producers of oxidative nor antioxidative markers. Our results provide a starting point of possible markers to better understand the disease pathogenesis in patients infected with T. gondii. Additional studies are needed to clarify the potential contribution of oxidative and antioxidative markers in these patients population.     

Vitamin E and beta-carotene status of dairy cows: a survey of plasma levels and supplementation practices

Culling rate in dairy cattle has increased considerably, thereby reducing cowś longevity and raising sustainability concerns worldwide. In the last decades, feeding systems have changed towards larger inclusion of preserved forages and reduced fresh herbage, which may limit vitamin E and beta-carotene dietary supply to dairy cows. Because of higher oxidative stress, engendered by greater milk production of modern genetics, the requirement for these nutrients is increased. Therefore, this study aimed to assess the current status of vitamin E and beta-carotene of commercial dairy cows. Blood vitamin E and beta-carotene concentrations were measured in 2 467 dairy cows from 127 farms in Belgium, Germany, Iberia and The Netherlands, that were visited once. Five cows were randomly selected per lactation stage per farm: Dry (between 30 and 1 day(s) before calving), Very-early (from calving until 15 days in milk (DIM)), Early (between 16 and 119 DIM), and Mid-late (from 120 DIM onwards). In addition, a survey was conducted to retrieve data on vitamin E and beta-carotene supplementation and feeding practices. Vitamin E and beta-carotene blood concentrations dropped considerably around calving. Among all surveyed cows, more than 75 and 44% were deficient in vitamin E and beta-carotene (i.e., blood concentration below 3.0 and 3.5 mg/l, respectively). Of the Very-early group, more than 97 and 78% of the cows were deficient in vitamin E and beta-carotene, respectively, with respective blood concentrations of 1.15 and 2.71 mg/l, which was significantly lower than the other lactation stages. Vitamin E and beta-carotene blood concentrations, as well as their supplementation levels, significantly varied among countries. Vitamin E and beta-carotene blood concentrations were positively related to the total estimated daily intakes of vitamin E and beta-carotene. Therefore, blood concentrations of vitamin E and beta-carotene depend on their respective level of intake, which is generally below recommendations and varies greatly between countries. Supplementation could contribute to provide cows with adequate amounts of vitamin E and beta-carotene all along the lactation, to ensure their lifetime performance and improve their fertility.     

Age-associated increase in PGE2 synthesis and COX activity in murine macrophages is reversed by vitamin E

We previouslyshowed that increased macrophage andPGE₂ production with age is due toenhanced cyclooxygenase (COX) activity and COX-2 expression. This studydetermined the effect of vitamin E supplementation on macrophagePGE₂ synthesis in young and old mice and its underlying mechanism. Mice were fed 30 or 500 parts permillion vitamin E for 30 days. Lipopolysaccharide (LPS)-stimulated macrophages from old mice produced significantly morePGE₂ than those from young mice.Vitamin E supplementation reversed the increasedPGE₂ production in old mice buthad no effect on macrophage PGE₂production in young mice. In both LPS-stimulated and unstimulated macrophages, COX activity was significantly higher in old than in youngmice at all intervals. Vitamin E supplementation completely reversedthe increased COX activity in old mice to levels comparable to those ofyoung mice but had no effect on macrophage COX activity of young miceor on COX-1 and COX-2 protein or COX-2 mRNA expression in young or oldmice. Thus vitamin E reverses the age-associated increase in macrophagePGE₂ production and COX activity.Vitamin E exerts its effect posttranslationally, by inhibiting COXactivity.

     

Effect of leucine metabolite beta -hydroxy-beta -methylbutyrate on muscle metabolism during resistance-exercise training

Nissen, S., R. Sharp, M. Ray, J. A. Rathmacher, D. Rice, J. C. Fuller, Jr., A. S. Connelly, and N. Abumrad. Effect of leucinemetabolite -hydroxy--methylbutyrate on muscle metabolism duringresistance-exercise training. J. Appl.Physiol. 81(5): 2095-2104, 1996.The effects ofdietary supplementation with the leucine metabolite-hydroxy--methylbutyrate (HMB) were studied in two experiments.In study 1, subjects(n = 41) were randomized among threelevels of HMB supplementation (0, 1.5 or 3.0 g HMB/day) and two proteinlevels (normal, 117 g/day, or high, 175 g/day) and weight lifted for1.5 h 3 days/wk for 3 wk. In study 2,subjects (n = 28) were fed either 0 or3.0 g HMB/day and weight lifted for 2-3 h 6 days/wk for 7 wk. Instudy 1, HMB significantly decreased the exercise-induced rise in muscle proteolysis as measured by urine3-methylhistidine during the first 2 wk of exercise (linear decrease,P < 0.04). Plasma creatinephosphokinase was also decreased with HMB supplementation(week 3, linear decrease,P < 0.05). Weight lifted wasincreased by HMB supplementation when compared with the unsupplementedsubjects during each week of the study (linear increase,P < 0.02). In study2, fat-free mass was significantly increased inHMB-supplemented subjects compared with the unsupplemented group at 2 and 4-6 wk of the study (P < 0.05). In conclusion, supplementation with either 1.5 or 3 g HMB/daycan partly prevent exercise-induced proteolysis and/or muscledamage and result in larger gains in muscle function associated withresistance training.

     

Dietary supplementation of pyrroloquinoline quinone disodium protects against oxidative stress and liver damage in laying hens fed an oxidized sunflower oil-added diet

The protective effects of dietary pyrroloquinoline quinone disodium (PQQ.Na₂) supplementation against oxidized sunflower oil-induced oxidative stress and liver injury in laying hens were examined. Three hundred and sixty 53-week-old Hy-Line Gray laying hens were randomly allocated into one of the five dietary treatments. The treatments included: (1) a diet containing 2% fresh sunflower oil; (2) a diet containing 2% thermally oxidized sunflower oil; (3) an oxidized sunflower oil diet with 100 mg/kg of added vitamin E; (4) an oxidized sunflower oil diet with 0.08 mg/kg of PQQ.Na₂; and (5) an oxidized sunflower oil diet with 0.12 mg/kg of PQQ.Na₂. Birds fed the oxidized sunflower oil diet showed a lower feed intake compared to birds fed the fresh oil diet or oxidized oil diet supplemented with vitamin E (P=0.009). Exposure to oxidized sunflower oil increased plasma malondialdehyde (P<0.001), hepatic reactive oxygen species (P<0.05) and carbonyl group levels (P<0.001), but decreased plasma glutathione levels (P=0.006) in laying hens. These unfavorable changes induced by the oxidized sunflower oil diet were modulated by dietary vitamin E or PQQ.Na₂ supplementation to levels comparable to the fresh oil group. Dietary supplementation with PQQ.Na₂ or vitamin E increased the activities of total superoxide dismutase and glutathione peroxidase in plasma and the liver, when compared with the oxidized sunflower oil group (P<0.05). PQQ.Na₂ or vitamin E diminished the oxidized sunflower oil diet induced elevation of liver weight (P=0.026), liver to BW ratio (P=0.001) and plasma activities of alanine aminotransferase (P=0.001) and aspartate aminotransferase (P<0.001) and maintained these indices at the similar levels to the fresh oil diet. Furthermore, oxidized sunflower oil increased hepatic DNA tail length (P<0.05) and tail moment (P<0.05) compared with the fresh oil group. Dietary supplementation of PQQ.Na₂ or vitamin E decreased the oxidized oil diet induced DNA tail length and tail moment to the basal levels in fresh oil diet. These results indicate that PQQ.Na₂ is a potential antioxidant and is as effective against oxidized oil-related liver injury in laying hens as vitamin E. The protective effects of PQQ.Na₂ against liver damage induced by oxidized oil may be partially due to its role in the scavenging of free radicals, inhibiting of lipid peroxidation and enhancing of antioxidant defense systems.     

Release of glutathione from erythrocytes and other markers of oxidative stress in carbon monoxide poisoning

Thom, Stephen R., Melissa Kang, Donald Fisher, and HarryIschiropoulos. Release of glutathione from erythrocytes and othermarkers of oxidative stress in carbon monoxide poisoning. J. Appl. Physiol. 82(5):1424-1432, 1997.Rats exposed to CO in a manner known to causeoxidative stress in brain exhibited a twofold increase in plasma levelsof oxidized proteins, thiobarbituric acid-reactive substances (TBARS),oxidized glutathione (GSSG), and reduced glutathione(GSH). Changes were neither directly related to hypoxicstress from carboxyhemoglobin nor significantly influenced bycirculating platelets or neutrophils. Treatment with the nitric oxidesynthase inhibitorN-nitro-L-arginine methylester inhibited elevations in GSH and GSSG but not changesin oxidized proteins or TBARS, suggesting that two oxidative mechanismsmay be operating in this model and that GSH and GSSG elevationsinvolved nitric oxide-derived oxidants. Elevations of blood GSH andGSSG occurred at different anatomic sites, indicating that no singleorgan was the source of the increased peptides. Animals that underwentexchange transfusion with a hemoglobin-containing saline solution didnot exhibit elevations in GSH and GSSG, suggesting that blood-bornecells released these peptides in response to oxidative stress. In invitro studies, erythrocytes, but not platelets and leukocytes,responded to oxidative stress from peroxynitrite by releasing GSH,whereas no release was observed in response to nitric oxide orsuperoxide. Glucose, maltose, and cytochalasin B, agents that protectextracellular components of the hexose transport protein complex fromoxidative stress, prevented GSH release. The data indicate that nitricoxide-derived oxidants are involved in CO-mediated oxidative stresswithin the vascular compartment and that elevations of severalcompounds may be useful for identifying exposures to CO likely toprecipitate brain injury.

     

Protein kinase C modulation of ventilatory response to hypoxia in nucleus tractus solitarii of conscious rats

This study aimedto determine the role of protein kinase C (PKC) in signal transductionmechanisms underlying ventilatory regulation in the nucleus tractussolitarii (NTS). Microinjection of phorbol 12-myristate 13-acetate intothe commissural NTS of nine chronically instrumented, unrestrained ratselicited significant cardiorespiratory enhancements that lasted for atleast 4 h, whereas administration of vehicle(n = 15) or the inactive phorbol ester 4-phorbol 12,13-didecanoate (n = 7)did not elicit minute ventilation (E)changes. Peak hypoxic Eresponses (10% O₂-balanceN₂) were measured in 19 additional animals after NTS microinjection of bisindolylmaleimide(BIM) I, a selective PKC inhibitor (n = 12), BIM V (inactive analog; n = 7),or vehicle (Con; n = 19). In Con,E increased from 139 ± 9 to 285 ± 26 ml/min in room air and hypoxia, respectively, and similarresponses occurred after BIM V. BIM I did not affect room airE but markedly attenuated hypoxia-induced E increases (128 ± 12 to 167 ± 18 ml/min; P < 0.02 vs. Con and BIM V). When BIM I was microinjected into the cerebellum(n = 4), cortex(n = 4), or spinal cord(n = 4),E responses were similar to Con.Western blots of subcellular fractions of dorsocaudal brain stemlysates revealed translocation of PKC, , , , , and  isoenzymes during acute hypoxia, and enhanced overall PKC activity wasconfirmed in the particulate fraction of dorsocaudal brain stem lysatesharvested after acute hypoxia. These studies suggest that, in the adultrat, PKC activation in the NTS mediates essential components of theacute hypoxic ventilatory response.

     

Estradiol protects cultured articular chondrocytes from oxygen-radical-induced damage

Osteoarthritis (OA) is aggravated in menopausal women possibly because of changed serum estrogen levels. Estradiol has been postulated to affect oxidative stress induced by reactive oxygen species (ROS) in articular chondrocytes. We generated ROS in cultured bovine articular chondrocytes by incubating them with combined Fe₂SO₄, vitamin C, and hydrogen peroxide. The release of thiobarbituric-acid-reactive substances (TBARS, lipid peroxidation) and lactate dehydrogenase (LDH, membrane damage) was measured photometrically. Various estradiol doses and vitamin E, serving as control with an established anti-oxidative capacity, were applied either upon each exchange of medium and during radical production (strategy 1) or only during radical production (strategy 2). In chondrocytes incubated according to strategy 1, the production of TBARS and LDH release were significantly suppressed by 10^–10–10^–4 M estradiol or by vitamin E. Under strategy 2, the production of TBARS was significantly suppressed at estradiol concentrations higher than 10^–6 M, whereas LDH release was inhibited at concentrations of 10^–6–10^–4 M. Vitamin E showed no significant effects. As repeated application of estradiol and vitamin E produced the best results, estradiol, like vitamin E, was speculated to accumulate in the plasma membrane and to decrease membrane fluidity resulting in protection against lipid peroxidation (non-genomic effect). Thus, in contrast to the neuroprotective effect of 17-estradiol in supraphysiological doses reported recently, the anti-oxidative potential of estradiol appears to protect articular chondrocytes from ROS-induced damage when the hormone is given repeatedly in a physiological range. Decreased estradiol levels may therefore contribute to menopausal OA in the long term.