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Biomedical subjects

I F Benzie

Publications and source records attributed to I F Benzie.

At least 19 recordsLinked to original sources
PubMed2001

Plasma ascorbic acid: measurement, stability and clinical utility revisited.

AIMS: To compare plasma ascorbic acid results by the colorimetric FRASC (Ferric Reducing/Antioxidant and Ascorbic Acid) assay and a reference HPLC method; to re-examine plasma ascorbic acid stability, and anticoagulant effect. DESIGN AND METHODS: For method comparison, 31 plasma samples were tested by both methods. For stability, matching EDTA, heparin, citrate and fluoride/oxalate plasma, stored under different conditions of time and temperature, was measured. RESULTS: FRASC is an acceptable alternative to HPLC for plasma ascorbic acid: precision, limit of detection and recovery were similar, and results by the two methods were indistinguishable: mean (95% CI) difference:1.8 (-1.1-4.6; n = 31) micromol/L. Ascorbic acid was most stable in heparinized plasma. Marked loss (p < 0.05) in EDTA plasma occurred within 30 min of blood collection. CONCLUSIONS: FRASC offers a speedy and reliable alternative to HPLC for plasma ascorbic acid. Heparin is proposed as the anticoagulant of choice; loss of ascorbic acid is rapid in EDTA plasma ex vivo.

Adult↗
PubMed2001

Water-soluble antioxidants in human tears: effect of the collection method.

PURPOSE: To resolve differences in published data on tear antioxidant levels by comparing the concentration of water-soluble antioxidants in human reflex tears collected by capillary tube and by the Schirmer strip collection method and in basal and reflex tears collected using the Schirmer strip method. METHODS: Yawn-induced reflex tears (collected simultaneously by capillary tubes and by Schirmer strips) and basal tears (by Schirmer strips and using local anesthetic) were collected from 12 healthy subjects. Tear cysteine, ascorbate, glutathione, urate, and tyrosine were measured by high-performance liquid chromatography within a few minutes of collection. RESULTS: Cysteine, ascorbate, glutathione, and tyrosine were 5 to 10 times higher (P < 0.01) in both reflex and basal tears collected by Schirmer strip compared with reflex tears collected by capillary tube from the same subject. Urate levels were slightly but nonsignificantly higher in Schirmer strip samples (P > 0.05). CONCLUSIONS: The conflict in published data on tear antioxidants is caused by differences in collection methods. With the exception of urate, antioxidants accumulate to very high levels in corneal cells. Spuriously high antioxidant levels in tears collected using Schirmer strips, therefore, are most probably caused by contamination with intracellular constituents. The capillary tube collection method is proposed as the method of choice for reflex tear collection for biochemical studies. This less-invasive method facilitates the evaluation of tear antioxidant levels as a biomonitoring tool for corneal health. Although moderately increased antioxidant levels may be beneficial, the authors hypothesize that marked increases may indicate damage to the ocular surface.

Adult↗
PubMed2000

Probe-assisted flow cytometric analysis of erythrocyte membrane response to site-specific oxidant stress.

BACKGROUND: Probe-assisted flow cytometry was used to monitor the response of membranes of living cells to oxidant stress in the presence and absence of antioxidants. Test conditions (fluorophore loading, oxidant concentration) were investigated and storage-related changes in erythrocyte response to oxidant stress explored. METHODS: Erythrocytes were incubated with a lipophilic fluorescent probe and exposed to site-specific oxidant challenge, induced by cumene hydroperoxide, in the presence and absence of urate, ascorbate, or alpha tocopherol in physiological amounts. Fluorescence of labeled and treated erythrocytes was measured for 120 min using a Coulter EPICS Elite ESP flow cytometer. RESULTS: Probe loading was dose and time dependent. Cumene hydroperoxide exhibited a potent and dose-dependent oxidant effect on erythrocyte membranes. Alpha tocopherol slowed, but did not prevent, membrane oxidation. Ascorbate appeared to have no effect on peroxidation initially, but then slowed and stopped propagation of membrane oxidation. The effect of urate was slight. CONCLUSIONS: This technique can provide insight into oxidative processes at the cellular level. Results indicated that lipophilic alpha tocopherol was the most effective antioxidant in slowing membrane peroxidation, but ascorbate appears to stop chain propagation. This effect may be owing to vitamin C/E interaction. Further study is needed.

Antioxidants↗
PubMed2000

Evolution of antioxidant defence mechanisms.

The metabolic strengths, weaknesses, opportunities and threats of the metabolic ability to split water brought about a proliferation of biological systems, produced a toxic oxygenic environment, and were responsible for the development of antioxidant defence mechanisms. Evolution is driven by heritable adaptations which improve environmental 'fit'. Hence aerobic respiration, using oxygen as a nutrient, came to predominate in biological systems, and antioxidant defence mechanisms which prevent and neutralise toxic oxygen intermediates have become widespread, varied, coordinated and effective. Antioxidant defences are not infallible however. In humans, reactive oxygen species-induced damage is associated with the ageing process, and with chronic diseases including cancer and coronary heart disease. Interestingly, some important antioxidants, including ascorbic acid and the tocopherols, cannot be synthesised by humans and must be taken in the diet. Another antioxidant, uric acid, is found in much higher concentrations in humans than in other mammals, and levels are also affected by diet. In humans, therefore, antioxidant defence against toxic oxygen intermediates is species specific and heavily influenced by nutrition. In this article, the atmospheric and metabolic changes which produced both the threat and opportunity offered by an oxygenic environment are outlined. An overview of oxygen toxicity, and adaptations to oxidative stress in terms of evolution of antioxidant defences, is presented. Finally, suggested benefits underlying our curious inability to manufacture ascorbic acid, and the possible role of uric acid in human antioxidant defence, are briefly discussed with particular reference to nutrition and toxicology.

Adaptation, Biological↗
PubMed2000

Interactions between vitamins C and E in human subjects.

Despite convincing in vitro evidence, a vitamin C-E interaction has not been confirmed in vivo. This study was designed to examine the effects of supplementation with either vitamin C or E on their respective plasma concentrations, other antioxidants, lipids and some haemostatic variables. Fasting blood was collected before and after intervention from thirty healthy adults in a double-blinded crossover study. Baselines for measured variables were established after 2 weeks of placebo supplementation, followed by daily supplementation with 73.5 mg RRR-alpha-tocopherol acetate or 500 mg ascorbic acid, and placebo, for 6 weeks. A 2 month washout preceded supplement crossover. Mean values showed that plasma lipid standardised alpha-tocopherol increased with ascorbic acid supplementation: from 4.09 (sem 0.51) to 4.53 (sem 0.66) micromol/mmol total cholesterol plus triacylglycerol (P < 0.05), and plasma ascorbic acid increased from 62.8 (sem 14.9) to 101.3 (sem 22. 2) micromol/l (P < 0.005). Supplementation with (RRR)-alpha-tocopherol acetate increased plasma alpha-tocopherol from 26.8 (sem 3.9) to 32.2 (sem 3.8) micromol/l (P < 0.05), and lipid-standardised alpha-tocopherol from 4.12 (sem 0.48) to 5.38 (sem 0.52) micromol/mmol (P < 0.001). Mean plasma ascorbic acid also increased with vitamin E supplementation, from 64.4 (sem 13.3) to 76. 4 (sem 18.4) micromol/l (P < 0.05). Plasma ferric reducing (antioxidant) power and glutathione peroxidase (U/g haemoglobin) increased in both groups, while urate, total cholesterol and triacylglycerol levels decreased (P < 0.05 throughout). Results are supportive of an in vivo interaction between vitamins C and E.

Adult↗
PubMed2000

Ascorbic acid concentration and total antioxidant activity of human tear fluid measured using the FRASC assay.

PURPOSE: To evaluate a novel method (FRASC) for total ferric reducing (antioxidant) activity and ascorbic acid concentration applied to human tears, to investigate the stability of ascorbic acid, and to determine the antioxidant status of human reflex tears. METHODS: Linearity, sensitivity, and precision of FRASC and ascorbic acid loss during 7 days' storage were assessed; total antioxidant activity and ascorbic acid and uric acid concentrations of reflex tears from 47 healthy subjects were measured. RESULTS: FRASC has good precision, linearity, and sensitivity. Ascorbic acid is stable for at least 7 days at moderately acidic pH (pH 3.6) and low temperature. Total antioxidant activity and ascorbic acid and uric acid concentrations (mean +/- SD) in reflex tears were 409 +/- 162, 23 +/- 9.6, and 68 +/- 46 microM, respectively. Ascorbic acid and uric acid constituted around half the total antioxidant activity measured. There was a significant correlation between uric acid and total antioxidant activity (r = 0.754; P: < 0.0001). Men had significantly (P: = 0.0045) higher tear ascorbic acid concentrations than women. CONCLUSIONS: FRASC is suitable for measuring total antioxidant activity and ascorbic acid in human tears. Further clinical study is needed to investigate the male-female difference seen, to characterize the remaining 50% antioxidant activity, and to investigate the effects of environmental conditions, antioxidant supplementation, age, and ocular disease on tear antioxidant status.

Adult↗
PubMed1999

"Antioxidant" (reducing) efficiency of ascorbate in plasma is not affected by concentration.

Ascorbate (vitamin C), an important dietary derived antioxidant, reportedly shows decreasing "antioxidant efficiency" with increasing concentrations in indirect radical trapping methods of antioxidant capacity. This study investigated the effect of concentration on antioxidant efficiency of ascorbate using a direct test of antioxidant capacity, the ferric reducing/antioxidant power test (FRAP assay). Results showed that the antioxidant efficiency factor of ascorbate was 2 and was constant over a wide concentration range in both plasma and pure aqueous solution. However, the absolute amount of ascorbate lost per unit of time increased with concentration. Furthermore, ascorbate was less stable in plasma than in aqueous solutions of similar pH and less stable in ethylenediamine tetraacetic acid (EDTA) than in heparinized plasma. Results indicate that previously reported concentration-dependent changes in antioxidant efficiency of ascorbate may have been caused by loss of ascorbate prior to and during testing, and by methodologic characteristics of indirect peroxyl radical trapping tests of antioxidant capacity. Therefore, it is suggested that the premise that the antioxidant efficiency of ascorbate is concentration-dependent is largely methodologically derived and does not reflect the antioxidant behavior of ascorbate per se.

Journal Article↗
PubMed1999

Vitamin C: prospective functional markers for defining optimal nutritional status.

Most species of plants and animals synthesize ascorbic acid, but human subjects cannot, making vitamin C an essential component of our diet. Relationships between vitamin C intake and status, and between status and health are not yet clear. There is evidence, however, that higher intake of vitamin C is associated with lower risk of disease, supporting the concept that optimal intake is needed for optimal vitamin C status, and that both factors are required for optimal health. Vitamin C has low toxicity in healthy subjects, but a clear definition of optimal status and the dietary intake required to meet and maintain this status is needed before a change in the current recommended intake can be considered. Available evidence suggests that intake of 200 mg vitamin C/d saturates tissues and maintains fasting plasma levels above the proposed threshold (50 mumol/l) for minimum risk of CHD. However, the issue of whether or not these levels produce 'optimal vitamin C status' awaits the clear and accepted definition of the term. This definition in turn awaits the development of reliable functional markers capable of assessing the effects of varying levels of vitamin C nutriture. In the present paper the relationship between intake and body stores of vitamin C and the role of vitamin C in human health are reviewed briefly. The requirements of a reliable functional marker of human vitamin C status are defined, three classes of functional markers (molecular, biochemical and physiological) are described, and possible candidate markers are examined.

Ascorbic Acid↗
PubMed1999

Total antioxidant capacity of teas by the ferric reducing/antioxidant power assay.

This study aimed to compare in vitro antioxidant power of different types of tea (Camellia sinensis). The ferric reducing/antioxidant power (FRAP) assay was used to measure the total antioxidant power of freshly prepared infusions of 25 types of teas. Results showed that different teas had widely different in vitro antioxidant power and that the antioxidant capacity was strongly correlated (r = 0. 956) with the total phenolics content of the tea. Expressed as micromol of antioxidant power/g of dried tea leaves, values ranged as 132-654 micromol/g for black ("fermented") teas, 233-532 micromol/g for Oolong ("semifermented") teas, and 272-1144 micromol/g for green ("nonfermented") teas. One cup of tea of usual strength (1-2%), therefore, can provide the same potential for improving antioxidant status as around 150 mg of pure ascorbic acid (vitamin C).

Antioxidants↗
PubMed1999

Interference of Hb-H disease in automated reticulocyte counting.

Hb-H disease is a form of alpha-thalassemia commonly found in south-east Asia. In this condition, numerous Hb-H bodies are found in erythrocytes using reticulocyte staining preparations. This is the first study addressing the possible interference of Hb-H inclusions in automated reticulocyte counting. In this study, seven Hb-H disease samples were tested. Results obtained by the visual method and the Technicon H*3 automated method agreed relatively well. This was in contrast to the Coulter STKS automated method which generally gave lower results. Furthermore, when the incubation time was extended to 180 min in the Coulter STKS method, two Hb-H disease samples gave results several-fold higher than those obtained using the 60 min incubation time. These discrepant results were likely to have been caused by the analyser using the wrong threshold to separate reticulocytes and mature erythrocytes. High and low interference in Hb-H disease samples was observed in the Coulter STKS automated reticulocyte method. Laboratories should be aware of this potential problem, particularly when samples are from patients of Asian origin.

Autoanalysis↗
PubMed1999

Consumption of green tea causes rapid increase in plasma antioxidant power in humans.

Green tea contains polyphenolic antioxidants that have shown anticarcinogenic properties in animal and in vitro experimental studies. Current data regarding absorption and bioavailability of tea antioxidants in humans, however, are conflicting. In this study, plasma and urine antioxidant power after ingestion of green tea was measured using the ferric reducing/antioxidant power (FRAP) assay (US patent pending) to assess absorption, systemic distribution, and renal excretion of green tea antioxidants in healthy adults. Results showed that absorption of green tea antioxidants was rapid, with peak increase in plasma FRAP of around 4% at 40 minutes after ingestion: mean increase was 44 +/- 9 (SE) mumol/l. Excretion of polyphenolic antioxidants was also fast, peaking at 60-90 minutes, with significant correlation between urinary FRAP values and urinary total phenolic concentrations (r = 0.845, p < 0.001). In control studies, no increase in plasma or urine FRAP values was seen after intake of water. Although the amount of antioxidants absorbed was relatively small and the increase in plasma antioxidant power was of short duration, results demonstrate that some potentially anticarcinogenic polyphenolic antioxidants in green tea enter the systemic circulation soon after ingestion and cause a significant increase in plasma antioxidant status. This increase may, in turn, lower oxidative damage to DNA and so decrease risk of cancer.

Adult↗
PubMed1998

Plasma ascorbate and vitamin E levels in Hong Kong Chinese.

OBJECTIVE: To describe fasting plasma total- and lipid standardised-vitamin E and ascorbate concentrations in Hong Kong Chinese subjects, and to explore age-, diet- and sex-related differences. DESIGN: Observational study. SETTING: Hong Kong. SUBJECTS: One hundred and fifty randomly selected Chinese subjects. INTERVENTIONS: Fasting plasma concentrations of ascorbate, total vitamin E and lipid standardised vitamin E (Vit E(LS), expressed as micromol vitamin E/mmol total cholesterol plus triglycerides) were measured. RESULTS: Total vitamin E concentrations ranged from 6-53 micromol/l (mean 24; s.d. 8.5), and Vit ELS from 0.85-7.09 micromol/mmol (mean 3.61; s.d. 1.19). Plasma Vit E(LS) concentrations in women were higher (P = 0.02) than in men: mean (median) concentrations 3.82 (3.88) and 3.37 (3.28) micromol/mmol, respectively. Vit ELS concentrations in men decreased with age. Mean (s.d.) plasma ascorbate concentrations in men and women were 49.7 (14.0) and 51.6 (21.6) micromol/l respectively, and did not show the significant age- and sex- differences reportedly found in Western studies. CONCLUSIONS: The hypothesis that Hong Kong Chinese, with their low risk of CHD, might have high Vit E(LS) levels in fasting plasma is not supported by the results obtained. Moreover, results showed male female differences and, in men, a previously unreported age-related decrease in plasma Vit E(LS) concentration which cannot be accounted for by concomitant differences in dietary intake of vitamin E alone. Results also suggest that the age- and sex- differences in plasma ascorbate seen in Western populations can be avoided by increased vitamin C intake.

Adult↗
PubMed1998

Antioxidant power of angiotensin-converting enzyme inhibitors in vitro.

AIMS: There is controversy regarding the potential antioxidant effect of captopril, therefore this study was performed to compare the in vitro antioxidant power of captopril with other angiotensin-converting enzyme (ACE) inhibitors. METHODS: Antioxidant power of captopril, enalapril, fosinopril, perindopril, quinapril and ramipril in aqueous solution was measured using the ferric reducing (antioxidant) power (FRAP) assay; captopril was also measured in ethanolic solution. RESULTS: Only captopril showed significant antioxidant power, demonstrating a stoichiometric factor of 1.0 in this assay. Concentration-related antioxidant power was seen in both aqueous and ethanolic solutions. CONCLUSIONS: Captopril shows antioxidant activity in vitro. This property could be relevant in vivo if captopril is concentrated in membranes, lipoproteins or at other important sites.

Angiotensin-Converting Enzyme Inhibitors↗