Impact of short-term bilberry supplementation on glycemic control, cardiovascular disease risk factors, and antioxidant status in Chinese patients with type 2 diabetes.

Bilberry (Vaccinium myrtillus L.) is one of the richest natural sources of anthocyanins which are powerful antioxidants and reported to have antiinflammatory, antidyslipidemic, antihypertensive, and hypoglycemic effects. The objective of this study was to assess the effect of bilberry supplementation on biomarkers of glycemic control, lipid profile, antioxidant, and inflammatory status in patients with type 2 diabetes in a randomized, double-blind, placebo-controlled cross-over study. Twenty patients were randomized to receive either bilberry supplementation (1.4 g/day of extract) daily for 4 weeks followed by 6 weeks of washout and then an additional 4 weeks of matching placebo or vice versa. Blood pressure, metabolic parameters, antioxidant status, and oxidative stress were measured before and after each period. Results showed no effect on body weight, blood pressure, or lipid profile. HbA1c was reduced by 0.31 ± 0.58% during bilberry supplementation, but this change was not significantly different from that with placebo. Antioxidant status, oxidative stress, and inflammatory status showed no significant differences across treatments. This short-term study of bilberry supplementation did not show significant effects on cardiovascular risk factors or antioxidant status, but the tendency for improved glycemic control may suggest a longer treatment period may be effective in diabetic patients.

Vitamin D deficiency, oxidative stress and antioxidant status: only weak association seen in the absence of advanced age, obesity or pre-existing disease.

Vitamin D deficiency (plasma 25-hydroxycholecalciferol (25(OH)D)70 % of participants were vitamin D deficient. No significant correlations and no biomarker differences across 25(OH)D quartiles or groups were seen except for total antioxidant status. A weak direct association (r 0·252, P<0·05) was observed between 25(OH)D and FRAP, and those in the lowest 25(OH)D quartile and group had significantly lower FRAP values. Results did not reveal a clear link between vitamin D status and oxidative stress biomarkers in the absence of advanced age, obesity and disease, though some evidence of depleted antioxidant status in those with vitamin D deficiency was seen. Poor antioxidant status may pre-date increased oxidative stress. Study of effects of correction of deficiency on antioxidant status and oxidative stress in vitamin D-deficient but otherwise healthy subjects is needed.

Vitamin D and oxidation-induced DNA damage: is there a connection?

Oxidation-induced damage to DNA can cause mutations, phenotypic changes and apoptosis. Agents that oppose such damage offer potential therapies for disease prevention. Vitamin D administration reportedly lowered DNA damage in type 2 diabetic mice, and higher DNA damage was reported in mononuclear cells of severely asthmatic patients who were vitamin D deficient. We hypothesised that lower vitamin D status associates with higher oxidation-induced DNA damage. Vitamin D deficiency (plasma 25(OH)D < 50 nmol/l) is highly prevalent worldwide, and association with DNA damage has high potential importance and impact in regard to the future health of vitamin D deficient young adults. In this study, oxidation-induced DNA damage in peripheral lymphocytes of 121 young (18-26 years) adults was measured using the formamidopyrimidine DNA glycosylase (FPG)-assisted comet assay. Plasma 25(OH)D was measured by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Correlational analysis was performed between 25(OH)D and DNA damage. Differences in DNA damage across tertiles of 25(OH)D were explored using analysis of variance. DNA damage in those with 25(OH)D <50 nmol and ≥50 nmol/l was compared using the unpaired t-test. Mean (SD) DNA damage (as %DNA in comet tail) and plasma 25(OH)D were, respectively, 18.58 (3.39)% and 44.7 (13.03) nmol/l. Most (82/121; 68%) of the subjects were deficient in vitamin D (25(OH)D <50nmol/l). No significant correlation was seen between 25(OH)D and DNA damage (r = -0.0824; P > 0.05). No significant difference was seen across 25(OH)D tertiles: mean (SD) %DNA in comet tail/25(OH)D nmol/l values in lowest, middle and highest tertiles were, respectively, 18.64 (3.30)/31.6 (4.4), 18.90 (3.98)/42.9 (3.5), 18.19 (2.84)/59.9 (8.5), nor across the binary divide: 18.73 (3.63)% in <50nmol/l group vs. 18.27 (2.84)% in the ≥50 nmol/l group. No association between vitamin D and oxidation-induced DNA damage was observed, but vitamin D deficiency was highly prevalent in the young adults studied, and we cannot rule out an ameliorative effect of correction of vitamin D deficiency on DNA damage.

Redox-linked effects of green tea on DNA damage and repair, and influence of microsatellite polymorphism in HMOX-1: results of a human intervention trial.

Green tea has many reported health benefits, including genoprotective and antioxidant effects, but green tea has pro-oxidant activity in vitro. A tea-induced pro-oxidant shift that triggers cytoprotective adaptations has been postulated, but human data are lacking. We investigated effects on oxidation-induced DNA damage and redox-linked cytoprotective factors, including 8-oxoguanine glycosylase (hOGG1) and heme oxygenase 1 (HMOX-1) in lymphocytes in a randomised, placebo-controlled, cross-over supplementation trial. hOGG1 catalyses the first step in base excision repair; increased HMOX-1 is a sign of cytoprotective response to pro-oxidant change. The influence of microsatellite polymorphisms in the HMOX-1 promoter region was also explored. Higher numbers of GT repeats [GT(n)] in this region reportedly diminish response to pro-oxidant change. Green tea [2 × 150 ml of 1% w/v tea/day (or water as control)] was taken for 12 weeks by 43 Type 2 diabetes subjects {20 with short [S/S; GT(n) < 25] and 23 with long [L/L; GT(n) ≥ 25]}. Fasting venous blood was collected before and after each treatment. The formamidopyrimidine DNA glycosylase-assisted comet assay was used to measure DNA damage in lymphocytes. For measuring hOGG1 activity, we used photo-damaged HeLa cells incubated with lymphocyte extracts from test subjects, in combination with the comet assay. Lymphocyte HMOX-1 and hOGG1 protein concentrations and expression (mRNA) of redox-sensitive genes, including HMOX-1 and hOGG1, were also investigated. Results showed significantly (P < 0.01) lower (~15%) DNA damage, higher (~50%) hOGG1 activity and higher (~40%) HMOX-1 protein concentration after tea. No changes in mRNA expression were seen. Baseline HMOX-1 protein and hOGG1 activity were higher (P < 0.05) in the S/S group, but tea-associated responses were similar in both GT(n) groups. Green tea is clearly associated with lowered DNA damage, increased hOGG1 activity and higher HMOX-1 protein levels. Further study is needed to confirm a cause and effect relationship and to establish if these effects are mediated by post-translational changes in proteins or by increased gene expression.

Desacyl ghrelin prevents doxorubicin-induced myocardial fibrosis and apoptosis via the GHSR-independent pathway.

Doxorubicin is an effective chemotherapeutic agent used to treat malignancies, but it causes cardiomyopathy. Preliminary evidence suggests that desacyl ghrelin might have protective effects on doxorubicin cardiotoxicity. This study examined the cellular effects of desacyl ghrelin on myocardial fibrosis and apoptosis in a doxorubicin cardiomyopathy experimental model. Adult C57BL/6 mice received an intraperitoneal injection of doxorubicin to induce cardiomyopathy, followed by 4-day treatment of saline (control) or desacyl ghrelin with or without [d-Lys3]-GHRP-6 (a growth hormone secretagogue receptor or GHSR1a antagonist). Ventricular structural and functional parameters were evaluated by transthoracic echocardiography. Molecular and cellular measurements were performed in ventricular muscle to examine myocardial fibrosis and apoptosis. Cardiac dysfunction was induced by doxorubicin, as indicated by significant decreases in ventricular fractional shortening and ejection fraction. This doxorubicin-induced cardiac dysfunction was prevented by the treatment of desacyl ghrelin no matter with or without the presence of [d-Lys3]-GHRP-6. Doxorubicin induced fibrosis (accumulated collagen deposition and increased CTGF), activated apoptosis (increased TUNEL index, apoptotic DNA fragmentation, and caspase-3 activity and decreased Bcl-2/Bax ratio), and suppressed phosphorylation status of prosurvival signals (ERK1/2 and Akt) in ventricular muscles. All these molecular and cellular alterations induced by doxorubicin were not found in the animals treated with desacyl ghrelin. Notably, the changes in the major markers of apoptosis, fibrosis, and Akt phosphorylation were found to be similar in the animals following the treatment of desacyl ghrelin with and without GHSR antagonist [d-Lys3]-GHRP-6. These findings demonstrate clearly that desacyl ghrelin protects the cardiomyocytes against the doxorubicin-induced cardiomyopathy by preventing the activation of cardiac fibrosis and apoptosis, and the effects are probably mediated through GHSR-independent mechanism.

Comparison of catechin profiles in human plasma and urine after single dosing and regular intake of green tea (Camellia sinensis).

Green tea (Camellia sinensis) catechin profiles in plasma and urine following single dosing and regular ingestion of green tea are not clear. We performed a placebo-controlled intervention study with sixteen healthy volunteers to determine changes in total and free catechins after a single dose and following 1 week of twice-daily green tea. Blood and urine samples were collected before (fasting) and after (60 and 120 min for blood; 90 and 180 min for urine) drinking 200 ml of 1.5% (w/v) green tea or water (n 8 each), and fasting samples were again collected after 7 d of 150 ml of 1% (w/v) supplemental green tea or water twice daily. After a 4-week washout, subjects were crossed onto the other treatment and procedures repeated. Plasma results at 1 h post-ingestion showed elevated (P < 0.05) mean epigallocatechin gallate (EGCG; 310 (SD 117) nmol/l; all in free form), epigallocatechin (EGC; 192 (SD 67) nmol/l; 30% free) and epicatechin gallate (ECG; 134 (SD 51) nmol/l; 75% free). Fasting plasma after 7 d of regular intake showed increased (P < 0.05) EGCG (80 v. 15 nmol/l at baseline) and ECG (120 v. 40 nmol/l), with > or =90% of both in their conjugated forms. Total EGC was < 10 nmol/l. Post-ingestion conjugation and renal loss of EGC and epicatechin were rapid and high, but were negligible for EGCG and ECG. In the green tea consumed, the content was EGCG > EGC > ECG, and the acute plasma response mirrored this. However, after chronic consumption there was almost no EGC found in fasting plasma, some EGCG was present, but a rather high level of ECG was maintained.

Study of potential cardioprotective effects of Ganoderma lucidum (Lingzhi): results of a controlled human intervention trial.

Previous studies have suggested that Lingzhi (Ganoderma lucidum) has antioxidant effects and possibly beneficial effects on blood pressure, plasma lipids and glucose, but these have not been confirmed in subjects with mild hypertension or hyperlipidaemia. The objective of the present study was to assess the cardiovascular, metabolic, antioxidant and immunomodulatory responses to therapy with Lingzhi in patients with borderline elevations of blood pressure and/or cholesterol in a controlled cross-over trial. A total of twenty-six patients received 1·44 g Lingzhi daily or matching placebo for 12 weeks in a randomised, double-blind, cross-over study with placebo-controlled run-in and cross-over periods. Body weight, blood pressure, metabolic parameters, urine catecholamines and cortisol, antioxidant status and lymphocyte subsets were measured after each period. Lingzhi was well tolerated and data from twenty-three evaluable subjects showed no changes in BMI or blood pressure when treated with Lingzhi or placebo. Plasma insulin and homeostasis model assessment-insulin resistance were lower after treatment with Lingzhi than after placebo. TAG decreased and HDL-cholesterol increased with Lingzhi but not with placebo in the first treatment period, but significant carry-over effects prevented complete analysis of these parameters. Urine catecholamines and cortisol, plasma antioxidant status and blood lymphocyte subsets showed no significant differences across treatments. Results indicate that Lingzhi might have mild antidiabetic effects and potentially improve the dyslipidaemia of diabetes, as shown previously in some animal studies. Further studies are desirable in patients with hyperglycaemia.

Increasing the antioxidant content of food: a personal view on whether this is possible or desirable.

A commonly held belief is that higher intake of antioxidants will promote better health through enhanced antioxidant status and lowered oxidative stress. However, the benefits of antioxidant-rich foods have not been reproduced in supplementation trials with pure antioxidants. This has driven research and commercial interest in foods, including traditional foods and their components, with enhanced antioxidant content and improved antioxidant bioavailability, which in many cases is very low. In this paper, evidence for the health benefits of antioxidant-rich foods and methods to increase the antioxidant content and bioavailability of food antioxidants are reviewed briefly, and the concept that increased food antioxidant content/intake per se is beneficial is examined from a cautionary perspective, considering issues of low bioavailability, rapid catabolism, biotransformation and the paradoxical pro-oxidant effects of dietary antioxidants.

Protective effect of caspase inhibition on compression-induced muscle damage.

There are currently no effective therapies for treating pressure-induced deep tissue injury. This study tested the efficacy of pharmacological inhibition of caspase in preventing muscle damage following sustained moderate compression. Adult Sprague-Dawley rats were subjected to prolonged moderate compression. Static pressure of 100 mm Hg compression was applied to an area of 1.5 cm2 in the tibialis region of the right limb of the rats for 6 h each day for two consecutive days. The left uncompressed limb served as intra-animal control. Rats were randomized to receive either vehicle (DMSO) as control treatment (n =8) or 6 mg kg⁻¹ of caspase inhibitor (z-VAD-fmk; n =8) prior to the 6 h compression on the two consecutive days.Muscle tissues directly underneath the compression region of the compressed limb and the same region of control limb were harvested after the compression procedure.Histological examination and biochemical/molecular measurement of apoptosis and autophagy were performed. Caspase inhibition was effective in alleviating the compression-induced pathohistology of muscle. The increases in caspase-3 protease activity, TUNEL index, apoptotic DNA fragmentation and pro-apoptotic factors (Bax, p53 and EndoG) and the decreases in anti-apoptotic factors (XIAP and HSP70) observed in compressed muscle of DMSO-treated animals were not found in animals treated with caspase inhibitor. The mRNA content of autophagic factors (Beclin-1, Atg5 and Atg12) and the protein content of LC3, FoxO3 and phospho-FoxO3 that were down-regulated in compressed muscle of DMSO-treated animals were all maintained at their basal level in the caspase inhibitor treated animals. Our data provide evidence that caspase inhibition attenuates compression-induced muscle apoptosis and maintains the basal autophagy level. These findings demonstrate that pharmacological inhibition of caspase/apoptosis is effective in alleviating muscle damage as induced by prolonged compression.

Habitual exercise increases resistance of lymphocytes to oxidant-induced DNA damage by upregulating expression of antioxidant and DNA repairing enzymes.

The underlying mechanisms of adaptation from staying physically active are not completely revealed. This study examined the effects of 8 and 20 weeks of habitual voluntary exercise on the susceptibility of lymphocytes to oxidant-induced DNA damage, antioxidant enzyme activities in cardiac and skeletal muscles, and circulatory antioxidant profile. Forty young adult rats were randomly assigned to sedentary control and exercise groups for an experimental period of 8 or 20 weeks. Animals assigned to exercise groups were subjected to 24 h daily free access to an in-cage running wheel with circumference of 1.19 m. A magnetic digital counter was attached to the running wheel to record daily exercise distance run by the animals. Control rats were housed in cages without a running wheel, located next to the exercised animals. Body weight and food intake were recorded weekly. After the experimental periods of 8 and 20 weeks, blood, left ventricle, soleus and plantaris muscles were collected for analysis. No significant difference was found in plasma total antioxidant capacity between exercised and control animals in the 8 and 20 week groups according to our ferric reducing/antioxidant power (FRAP) analysis. However, modified FRAP for ascorbic acid (FRASC) analysis indicated that plasma ascorbic acid content was significantly increased by 46 and 34% in 8 and 20 week exercise groups, respectively, when compared with the corresponding control groups. Superoxide dismutase (SOD) activity was significantly elevated by 39% in erythrocytes of animals exercised for 8 weeks relative to control animals. In the 20 week exercise group, Glutathione peroxidase (GPx) activity in ventricle and plantaris was significantly upregulated by 477 and 290%, respectively, relative to control values. As demonstrated by comet assay, the oxidant-induced DNA damage was significantly reduced by 21 and 45% in lymphocytes of animals exercised for 8 and 20 weeks, respectively, when compared with the corresponding control lymphocytes. Our qRT-PCR analysis showed that the transcript expression of SOD2 was significantly elevated by 939% in lymphocytes of animals exercised for 8 weeks relative to control animals. Increased expressions of SOD2 (by 19%), catalase (25%), APEX nuclease (multifunctional DNA repair enzyme) 1 (APEX1; 46%), Protein kinase, DNA-activated, catalytic polypeptide (Prkdc; 9%) and O-6-methylguanine-DNA methyltransferase (Mgmt; 26%) were found in lymphocytes of animals exercised for 20 weeks relative to control rats. These results demonstrate that habitual exercise confers increased resistance of lymphocytes to oxidant-induced DNA damage, and this protective effect is possibly attributed to the regular exercise-induced elevated expression of antioxidant and DNA repairing enzymes.

Genoprotective effects of green tea (Camellia sinensis) in human subjects: results of a controlled supplementation trial.

Green tea is rich in polyphenolic antioxidants and has widely reported but largely unsubstantiated health benefits. In the present study, genoprotective effects of two types of green tea were studied both in an in vitro and in a human supplementation trial. For the in vitro study, human lymphocytes were pre-incubated in tea (0·005-0·1 %, w/v), washed and subjected to oxidant challenge induced by H2O2. In a placebo-controlled, cross-over supplementation study, eighteen healthy volunteers took 2 x 150 ml/d of 1% (w/v) green tea ('Longjing' green tea or 'screw-shaped' green tea) or water (control) for 4 weeks (n 6). Subjects took all the three treatments in a random order, with 6 weeks' washout between each treatment. Fasting blood and urine were collected before and after each treatment. The comet assay was used to measure the resistance of lymphocytic DNA to H2O2-induced challenge. Basal oxidation-induced DNA damage was measured using the formamidopyrimidine glycosylase (Fpg) enzyme-assisted comet assay. Urine 7,8-dihydro-2-deoxyguanosine (8-oxodG, mol/mmol creatinine), a biomarker of whole-body oxidative stress, was measured by liquid chromatography with tandem MS. In vitro testing results of tea-treated cells showed increased (P < 0·05) resistance of DNA to the challenge. In the supplementation trial, a significant (P < 0·05) increase in resistance was also observed. Furthermore, the FPg comet data showed .20% decrease in DNA damage with tea supplementation: mean and standard deviation changes in %DNA in comet tail in the Fpg-assisted comet assay were: -5·96 (SD 3·83) % after Longjing tea; -6·22 (SD 3·34) % after screw-shaped tea; +0·91 (SD 5·79) % after water (P < 0·05). No significant changes in urine 8-oxodG were seen. The results indicate that green tea has significant genoprotective effects and provide evidence for green tea as a 'functional food'.

Antioxidant content and ultraviolet absorption characteristics of human tears.

PURPOSE: Dry eye syndrome is a common age-related disorder, and decreased antioxidant/ultraviolet (UV) radiation protection in tears may be part of the cause. This study aimed to compare the tear antioxidant content and flow rate in young and older adults. The total antioxidant content and UV absorbing properties of various commercially available ophthalmic solutions used to alleviate dry eye symptoms were also examined. METHODS: Minimally stimulated tears were collected from 120 healthy Chinese adults with no ocular pathology. Two age groups were studied: 19 to 29 years (n = 58) and 50 to 75 years (n = 62). Tear samples from each subject and 13 ophthalmic solutions were analyzed for total antioxidant content (as the Ferric Reducing/Antioxidant Power value). Tear flow rates were estimated from time taken to collect a fixed volume of tear fluid. UV absorbance spectra of pooled fresh reflex tear fluid and the ophthalmic solutions were determined. RESULTS: Results showed that the antioxidant content of minimally stimulated tears from older subjects (398 ± 160 µmol/l) was not significantly lower than that of younger subjects (348 ± 159 µmol/l; p = 0.0915). However, there was a significant difference in the tear flow rates between the two groups (p < 0.0001), with the younger group having three to four fold higher flow rate. None of the commercial preparations tested had detectable antioxidant content, and none showed the UV absorption characteristics of natural reflex tears. CONCLUSIONS: The effect of low flow rate on the dynamic antioxidant supply to the corneal surface indicates that older subjects have poorer overall defense against photooxidative and other oxidative processes. This could predispose older persons to corneal stress and development of dry eye syndrome. The commercially available artificial tears tested lack both the antioxidant content and UV absorbing characteristics of natural tears. Artificial tears formulations that help restore natural antioxidant and UV absorbing properties to the tear film of the aging eye may help prevent or improve dry eye symptoms and promote ocular health.

Relationships among diabetic retinopathy, antioxidants, and glycemic control.

PURPOSE: Type 2 Diabetes Mellitus (DM) is increasing worldwide and affects ∼11% of the Hong Kong population. Diabetic retinopathy (DR) is a common cause of vision loss in type 2 DM. Risk of DR is increased by poor glycemic control, elevated lipids, and blood pressure, but it is not possible to predict the development or progression of DR at an individual level. Increased oxidative stress is thought to play a role. The use of a wider biomarker profile incorporating biomarkers of antioxidant status and oxidative stress may aid identification of individuals at higher risk or at very early stages of developing DR. METHODS: Four hundred twenty type 2 DM subjects without diabetic complications were investigated. Eyes were examined for DR and anterior and posterior ocular segment pathology. DR was graded according to Early Treatment Diabetic Retinopathy Study criteria. Demographic data were collected. Traditional risk factors plus biomarkers of antioxidant status and oxidative stress in fasting blood and urine were determined. RESULTS: Overall DR prevalence was 89%. No significant differences in any demographic measures or biomarkers were found among those subjects with different DR grades, or in those without DR. Significant correlations (p < 0.0001) between HbA1c and DNA damage, (ρ = 0.32) and fasting plasma glucose and DNA damage (ρ = 0.52) were seen. DNA damage was also significantly and inversely correlated (p < 0.0001) with both plasma ascorbic acid (ρ = -0.41) and plasma total antioxidant level (ρ = -0.21). CONCLUSIONS: DR prevalence was very high in this group, but no biomarker differences were seen in those with DR compared to those free of DR, or in those with different degrees of severity of DR. This group of 420 subjects is being followed up to investigate whether the extended biomarker profile at baseline is related to progression of and/or incident DR.

DNA protective effect of ginseng and the antagonistic effect of Chinese turnip: a preliminary study.

The genoprotective effect of American and Asian ginseng on human lymphocytic DNA was studied. Using the comet assay, aqueous extracts of both types of ginseng were shown to diminish hydrogen peroxide-induced DNA damage. In contrast, and in accordance with traditional Chinese medicine beliefs, addition of the juice from Chinese turnip counteracted the beneficial effect of ginseng. Results showed that incubating ginseng along with turnip juice abolished the DNA protective effect of both American and Asian ginseng. Although the exact mechanism has not been elucidated, the counteracting effect of turnip on ginseng seems unlikely to be mediated by enzymatic action as the effect was seen with boiled as well as unboiled turnip extract.

Effects of incense smoke on human lymphocyte DNA.

Incense burning is common in Southeast Asia, where it is a traditional and ceremonial practice in deity worship and paying respect to ancestors. However, incense emissions are an important source of indoor air pollution in Asia, and may induce health problems to those exposed. In this in vitro study the effects of incense emissions on human DNA were investigated using the comet assay. Particulates in smoke from six kinds of incense were trapped in saline or ethanol and human lymphocytes were exposed under controlled conditions. Results showed that DNA damage, including strand breaks, was induced by both aqueous and ethanolic extracts of two samples. The ethanolic extract of one sample induced DNA damage, while no significant DNA damage was found in the remaining three samples. The mechanisms underlying DNA damage induced by incense emissions were also investigated. Catalase (CAT), sodium azide, and superoxide dismutase (SOD) were co-incubated with extract, which exerted significant DNA damaging effects. Results showed that CAT with or without SOD diminished DNA damage, whereas sodium azide did not seem able to reduce DNA damage. Data indicate there are potential adverse health effects of such exposure, particularly for temple workers.

Do multipurpose solutions damage porcine corneal epithelial cells?

PURPOSE: To determine and compare the cytotoxic effects on porcine corneal epithelial cells of commercially available multipurpose solutions (MPS) using fluorescein staining and flow cytometry (FCM). METHODS: Effects of exposure time of 10 s to 10 min of MPS containing polyquaternium-1 (MPS-A), polyaminopropyl biguanide (MPS-B), and polyhexanide (MPS-C), on porcine corneal epithelial cells were determined. Cell viability and membrane integrity were assessed by Annexin V-FITC/7-AAD staining and FCM. In further trials, whole corneas were immersed in MPS and control (buffered saline), and corneal fluorescein staining assessed before FCM analysis. RESULTS: Significantly higher percentages of 7-AAD-stained cells (early necrosis) were observed at all exposure times for MPS-A than for other solutions and control (p < 0.05). Exposure time in MPS-A and 7-AAD-stained cell proportions showed significant correlation (r = 0.9957; p < 0.0001). Significantly more cells dual-stained with Annexin V-FITC/7-AAD (late necrosis) after 5 min MPS-A exposure (11.8 +/- 1.1%), compared with 1.2 +/- 0.9% (MPS-B), 0.9 +/- 0.5% (MPS-C), and 1.8 +/- 0.2% (control). However, only 10 min exposure resulted in significant increases in fluorescein grades (p < 0.001), with median grade 0.75 for MPS-A, and 0.50 for the other MPS. CONCLUSIONS: MPS exposure, especially MPS-A, affected the viability and integrity of porcine corneal epithelial cells. Furthermore, our results confirmed that fluorescein staining correlates poorly with cytotoxicity. As fluorescein staining lacks sensitivity to determine cytotoxic effects of ophthalmic solutions, more objective and sensitive assessment methods such as differential staining and FCM should be developed.

Acute hyperglycemia and oxidative stress: direct cause and effect?

Oxidative stress is increased in Type 2 diabetes and this appears to underlie the development of diabetic complications. Increased oxidative stress is claimed to be triggered directly by acute (sudden-onset) hyperglycemia, but published data do not clearly support a direct cause and effect relationship. In this article, published evidence of a direct prooxidant effect of acute hyperglycemia is presented and discussed in some detail, and conflicts, controversies, and problems are highlighted. Evidence for glucose variability as a possible important trigger of oxidative stress in diabetes is reviewed, with some speculation as to how the field would be advanced if there were more widespread recognition about the role that wide fluctuations in glucose concentration play in diabetic complications. Possible direct or indirect antioxidative effects of various drugs used in the treatment of diabetic subjects are discussed because these may have influenced current understanding of the link between hyperglycemia and oxidative stress. The aims are to reveal the divergence between the available evidence and the accepted view that acute hyperglycemia is a direct trigger of oxidative stress and to suggest areas of research that will help resolve current controversies in this important and challenging area.

Is the yin-yang nature of Chinese herbal medicine equivalent to antioxidation-oxidation?

It has been suggested that yin-yang theory described in traditional Chinese medicine is somewhat equivalent to the modern theory of antioxidant-oxidant balance. Some yin-tonic Chinese herbal medicines possess antioxidant properties. In this context, the DNA protective effect of 12 yin-tonic and 13 yang-tonic herbs were tested using the single cell gel electrophoresis (comet) assay. Lymphocytes from three healthy subjects were pre-incubated with aqueous herb extract, and the comet assay was performed on treated, untreated, challenged and unchallenged cells in parallel, oxidant challenge being induced by 5 min exposure to hydrogen peroxide. Results using this ex vivo cellular assay showed protection by some herbs. Seven out of 12 yin-tonic Chinese herbs demonstrated decreased DNA damage after treatment while 10 out of 13 yang-tonic herbs showed protection. Among 25 herbs tested, rhizome of Ligusticum sinensis Oliv. and aerial part of Artemisia annua L. demonstrated greatest DNA protective effect. Results indicated that the yin nature of herbs may not be necessarily associated with superior antioxidative effect to yang-tonic herbs, at least in terms of DNA protection against oxidant challenge.

Affective images: relieving chronic pain and enhancing quality of life for older persons.

With increasing life expectancy, the incidence of chronic illness and chronic pain also increase. Chronic pain robs older people of their quality of life, and limits functional mobility and ambulation, which leads to muscle atrophy. Older people are often reluctant to request pain relief, attempting to endure pain as a "normal" part of ageing. Innovative non-pharmacological interventions in pain management are appealing. In this study, affective images and pictures were used in 15 elderly patients (12 female and three male) in chronic pain who needed to perform stretching and standing exercises in their physiotherapy sessions. Pain scores were measured by the Visual Analogue Scale (VAS), and health-related quality of life was measured by the Medical Outcomes Study Short Form 36 (SF-36). VAS and SF-36 were administered in weeks 1 and in 6. Pain scores were measured during physiotherapy session in the following weeks. There was a significant decreased in VAS from week 1 to week 6 (t = 3.607; df = 14; p < 0.05). Also, a significant decrease in VAS occurred when the patients were watching affective pictures while performing the physiotherapy exercise in weeks 2 and 3, but not in weeks 4 and 5. SF-36 increased in week 6, which indicated an increase in health-related quality of life, though not statistically significant. Affective images and pictures appear to be an effective non-pharmacological intervention in pain management for the older person.

The effect of music therapy on postoperative pain, heart rate, systolic blood pressures and analgesic use following nasal surgery.

The prevalence of unrelieved postoperative pain is high and may lead to adverse effects including prolonged hospitalization and delayed recovery. Distraction may be an effective pain-relieving strategy, and can be implemented by several means including affective imaging, games, and possibly music. The aim of this study was to explore the effect of music therapy on postoperative pain. Fifty-seven patients (24 females, 33 males; mean +/- SD age 39.9 +/- 14.35 years [range 15 to 69 years] were matched for age and sex and then nonselectively assigned to either an experimental (n = 27) or a control (n = 30) group. Music was played intermittently to members of the experimental group during the first 24 hour postoperative period. Pain intensity was measured using the Pain Verbal Rating Scales (VRS). Significant decreases in pain intensity over time were found in the experimental group compared to the control group (p < 0.0001). In addition, the experimental group had a lower systolic blood pressure and heart rate, and took fewer oral analgesics for pain. These findings suggest that music therapy is an effective nonpharmacologic approach for postoperative pain management.