Screening of Phosphodiesterase-5 Inhibitors and Their Analogs in Dietary Supplements by Liquid Chromatography-Hybrid Ion Trap-Time of Flight Mass Spectrometry.

An accurate and reliable method based on ion trap-time of flight mass spectrometry (IT-TOF MS) was developed for screening phosphodiesterase-5 inhibitors, including sildenafil, vardenafil, and tadalafil, and their analogs in dietary supplements. Various parameters affecting liquid chromatographic separation and IT-TOF detection were investigated, and the optimal conditions were determined. The separation was achieved on a reversed-phase column under gradient elution using acetonitrile and water containing 0.2% acetic acid at a flow rate of 0.2 mL/min. The chromatographic eluents were directly ionized in the IT-TOF system equipped with an electrospray ion source operating in the positive ion mode. The proposed screening method was validated by assessing its linearity, precision, and accuracy. Sequential tandem MS was conducted to obtain structural information of the references, and the fragmentation mechanism of each reference was proposed for providing spectral insight for newly synthesized analogs. Structural information, including accurate masses of both parent and fragment ions, was incorporated into the MSn spectral library. The developed method was successfully applied for screening adulterated dietary supplement samples.

Effects of glutathione s-transferase (GST) M1 and T1 polymorphisms on antioxidant vitamins and oxidative stress-related parameters in Korean subclinical hypertensive subjects after kale juice (Brassica oleracea acephala) supplementation.

BACKGROUND/OBJECTIVES: Glutathione s-transferase (GST) is involved in the formation of a multigene family comprising phase II detoxification enzymes, involved in the detoxification of reactive oxygen species. This study evaluated whether daily supplementation with kale juice could modulate levels of plasma antioxidant vitamins and oxidative stress-related parameters. We further examined whether this modulation was affected by combined GSTM1 and T1 polymorphisms. SUBJECTS/METHODS: Totally, 84 subclinical hypertensive patients having systolic blood pressure (BP) over 130 mmHg or diastolic BP over 85 mmHg, received 300 mL of kale juice daily for 6 weeks. Blood samples were drawn before start of study and after completion of 6 weeks. RESULTS: After supplementation, we observed significant decrease in DNA damage and increase in erythrocyte catalase activity in all genotypes. Plasma level of vitamin C was significantly increased in the wild/null and double null genotypes. The plasma levels of ß-carotene, erythrocyte glutathione peroxidase activity, and nitric oxide were increased only in the wild/null genotype after kale juice supplementation. CONCLUSIONS: The effect of kale juice was significantly greater in the GSTM1 null genotype and wild/null genotype groups, suggesting possibility of personalized nutritional prescriptions based on personal genetics.

Purple grape juice supplementation in smokers and antioxidant status according to different types of GST polymorphisms.

DNA damages and antioxidant status was assessed after 8 weeks of purple grape juice supplementation in male smokers depending on the glutathione S-transferase polymorphisms. Ninety-five smokers consumed 480 ml of purple grape juice for 8 weeks. The blood samples were collected before and after supplementation to measure lymphocyte DNA damages, plasma antioxidants, conjugated diene, and the erythrocyte antioxidant enzymes. The diastolic pressure, lymphocyte DNA damage, and plasma conjugated diene were significantly decreased but the plasma γ-tocopherol was increased in GSTM1-null genotype, while increased blood glutathione and decreased lymphocyte DNA damage were observed in GSTM1-present genotype. In case of GSTT1 on the other hand, the decrease in diastolic pressure and lymphocyte DNA damage was observed in both null types and present types, but the erythrocyte catalase activity was decreased in GSTT1-null type and the plasma vitamin C level was increased in GSTT1-present type, suggesting that, the antioxidant effect of grape juice was greater in GSTT1-present type compared to GSTT1-null type. The intakes of 8-week purple grape juice affected diastolic blood pressures, DNA damage reductions and antioxidant status in smokers, mainly greater in GSTM1-null type and GSTT1-present type.

Association between kimchi intake and asthma in Korean adults: the fourth and fifth Korea National Health and Nutrition Examination Survey (2007-2011).

This study was performed to investigate the relationship between dietary factors and asthma in a representative population-based sample of 19,659 men and women, aged 19-64 years, using data from the fourth and fifth Korean National Health and Nutrition Examination Survey (KNHANES), 2007-2011. The presence of asthma was based on self-reported physician diagnosis of asthma in the Health Interview Surveys. Food intake was estimated by trained interviewers using a 24-h recall method. The prevalence of asthma in Korean adults was 2.4%. Adults with asthma consumed fewer amounts of kimchi (P=.0444) and fish (P=.0175) but had a higher cereal intake than those without asthma (P=.0056). Multiple logistic regression analysis after controlling for confounding factors showed a significant inverse relationship between kimchi consumption and the prevalence of asthma [odds ratio (95% confidence interval) for subjects consuming 1 to <2 servings (40-79.9 g), 2 to <3 servings (80-119.9 g), and ≥3 servings (≥120 g), relative to those consuming <1 serving (<40 g): 0.726 (0.534-0.987), 0.506 (0.348-0.736), and 0.678 (0.502-0.916), respectively; P for trend=0.0131]. These results warrant future studies to explore the mechanisms responsible for the association between kimchi consumption and asthma.

Folic acid supplementation reduces oxidative stress and hepatic toxicity in rats treated chronically with ethanol.

Folate deficiency and hyperhomocysteinemia are found in most patients with alcoholic liver disease. Oxidative stress is one of the most important mechanisms contributing to homocysteine (Hcy)-induced tissue injury. However it has not been examined whether exogenous administration of folic acid attenuates oxidative stress and hepatic toxicity. The aim of this study was to investigate the in vivo effect of folic acid supplementation on oxidative stress and hepatic toxicity induced by chronic ethanol consumption. Wistar rats (n = 32) were divided into four groups and fed 0%, 12%, 36% ethanol, or 36% ethanol plus folic acid (10 mg folic acid/L) diets. After 5 weeks, chronic consumption of the 36% ethanol diet significantly increased plasma alanine transaminase (ALT) (P < 0.05) and aspartate transaminase (AST) (P < 0.05), triglycerides (TG) (P < 0.05), Hcy (P < 0.001), and low density lipoprotein conjugated dienes (CD) (P < 0.05) but decreased total radical-trapping antioxidant potential (TRAP) (P < 0.001). These changes were prevented partially by folic acid supplementation. The 12% ethanol diet had no apparent effect on most parameters. Plasma Hcy concentration was well correlated with plasma ALT (r = 0.612(**)), AST (r = 0.652(*)), CD (r = 0.495(*)), and TRAP (r = -0.486(*)). The results indicate that moderately elevated Hcy is associated with increased oxidative stress and liver injury in alcohol-fed rats, and suggests that folic acid supplementation appears to attenuate hepatic toxicity induced by chronic ethanol consumption possibly by decreasing oxidative stress.

The effect of carrot juice, ß-carotene supplementation on lymphocyte DNA damage, erythrocyte antioxidant enzymes and plasma lipid profiles in Korean smoker.

High consumption of fruits and vegetables has been suggested to provide some protection to smokers who are exposed to an increased risk of numerous cancers and other degenerative diseases. Carrot is the most important source of dietary ß-carotene. Therefore, the objective of this study was to investigate whether carrot juice supplementation to smokers can protect against lymphocyte DNA damage and to compare the effect of supplementation of capsules containing purified ß-carotene or a placebo (simple lactose). The study was conducted in a randomized and placebo-controlled design. After a depletion period of 14 days, 48 smokers were supplemented with either carrot juice (n = 18), purified ß-carotene (n = 16) or placebo (n = 14). Each group was supplemented for 8 weeks with approximately 20.49 mg of ß-carotene/day and 1.2 mg of vitamin C/day, as carrot juice (300 ml/day) or purified ß-carotene (20.49 mg of ß-carotene, 1 capsule/day). Lymphocyte DNA damage was determined using the COMET assay under alkaline conditions and damage was quantified by measuring tail moment (TM), tail length (TL), and% DNA in the tail. Lymphocyte DNA damage was significantly decreased in the carrot juice group in all three measurements. The group that received purified ß-carotene also showed a significant decrease in lymphocyte DNA damage in all three measurements. However, no significant changes in DNA damage was observed for the placebo group except TM (P = 0.016). Erythrocyte antioxidant enzyme was not significantly changed after supplementation. Similarly plasma lipid profiles were not different after carrot juice, ß-carotene and placebo supplementation. These results suggest that while the placebo group failed to show any protective effect, carrot juice containing beta-carotene or purified ß-carotene itself had great antioxidative potential in preventing damage to lymphocyte DNA in smokers.

Sociodemographic and lifestyle factors are associated with the use of dietary supplements in a Korean population.

OBJECTIVE: The use of dietary supplements has been increasing rapidly in Korea over the last decade. The aim of this study was to investigate associations between the pattern of dietary supplement use and the sociodemographic/lifestyle characteristics of Korean consumers. METHODS: Participants were asked to complete a self-administered questionnaire on their sociodemographic and lifestyle characteristics, perceived health status, and regular dietary supplement use. RESULTS: A total of 697 men and 832 women completed the questionnaire. Of the respondents, 44.3% of the men and 53.2% of the women used some kind of dietary supplement regularly. Dietary supplement users were more likely to be women (P < 0.001), to be older than 50 years (P < 0.001), to have a higher household income (P = 0.003), to engage in moderate or vigorous physical activity (P < 0.032), to perceive themselves as healthy (P = 0.026), and to have received a diagnosis of a chronic disease (P < 0.001). In addition, the type of dietary supplements used varied with respect to sociodemographic and lifestyle factors. Among dietary supplement users, men preferred ginseng, and older respondents were more likely to use carbohydrate supplements and less likely to use lipid supplements. Those who had a lower BMI, were ex-smokers, or were nondrinkers preferred either vitamins or minerals. Those who were highly physical active or were nondrinkers tended to prefer either vitamin/mineral complexes or carbohydrate supplements. CONCLUSIONS: The use of dietary supplements was related to sociodemographic and lifestyle factors in a Korean population.

Six-week supplementation with Chlorella has favorable impact on antioxidant status in Korean male smokers.

OBJECTIVE: Chlorella vulgaris is a popular food supplement in Asia and is currently marketed as a nutritional supplement. However, available scientific studies do not support its effectiveness for preventing or treating any disease in humans. Because Chlorella contains numerous nutrients, including antioxidants, it is thought to exert antioxidative functions by scavenging free radicals created by various environmental factors such as smoking. The purpose of this study was to investigate whether 6 wk of Chlorella supplementation to smokers is protective against oxidative damage in a randomized, double-blinded, placebo-controlled trial. METHODS: Fifty-two smokers, aged 20-65 y, were given 6.3g of Chlorella or placebo every day for 6 wk. Blood samples were drawn at the beginning and after the supplementation. Plasma antioxidant vitamin levels and lipid peroxidation levels were measured. As a marker of oxidative stress, lymphocyte DNA damage was measured. RESULTS: Chlorella supplementation increased plasma vitamin C (44.4%), alpha-tocopherol (15.7%), and erythrocyte catalase and superoxide dismutase activities. Although 6 wk of Chlorella supplementation resulted in a significant decrease in lymphocyte DNA damage, as measured by comet assay, placebo supplementation also decreased the measured amount of lymphocyte DNA damage. CONCLUSION: Chlorella supplementation resulted in the conservation of plasma antioxidant nutrient status and improvement in erythrocyte antioxidant enzyme activities in subjects. Therefore, our results are supportive of an antioxidant role for Chlorella and indicate that Chlorella is an important whole-food supplement that should be included as a key component of a healthy diet.

Induction of apoptosis in human colon cancer HCT-116 cells by anthocyanins through suppression of Akt and activation of p38-MAPK.

Anthocyanins belong to a class of flavonoids that exhibit important anti-oxidant and anti-inflammatory actions as well as chemotherapeutic effects. However, little is known concerning the molecular mechanisms by which these activities are exerted. In this study, we investigated the anthocyanins isolated from Vitis coignetiae Pulliat for their potential anti-proliferative and apoptotic effects on human colon cancer HCT-116 cells. These anthocyanins inhibited cell viability and induce apoptotic cell death of HCT-116 cells in a dose-dependent manner. The apoptotic cell death was caspase-dependent and the anthocyanins regulated anti-apoptotic proteins (IAPs). In addition, apoptosis was associated with activation of p38-MAPK and suppression of Akt. In conclusion, this study suggests that the anthocyanins isolated from Vitis coignetiae Pulliat induce apoptosis might at least in part through activating p38-MAPK and suppressing Akt in human colon cancer HCT-116 cells.

Changes in antioxidant status, blood pressure, and lymphocyte DNA damage from grape juice supplementation.

In a placebo-controlled double-blinded study, the effect of 8 weeks of grape juice was compared to the effect of isocaloric placebo juice. Volunteers with a systolic blood pressure > 130 mmHg and diastolic blood pressure > 90 mmHg were recruited. A total of 40 healthy subjects were randomized to receive isocaloric juices for 8 weeks. Twenty-one subjects were instructed to consume 5.5 mL/kg daily of grape juice (GJ), and 19 subjects consumed placebo juice (PJ). Plasma antioxidant vitamin C, total radical trapping antioxidant capacity, blood pressure, and lymphocyte DNA damage were assessed pre- and postsupplementation. Plasma total radical-trapping antioxidant potential showed an increase at the level of 1.31 +/- 0.01 (postsupplementation) versus 1.33 +/- 0.01 (presupplementation) (P < 0.1). Grape juice consumption resulted in a 26% decrease in lymphocyte DNA (both hydrogen peroxide treated or spontaneous) in the grape juice group, while no difference was found in the PJ group. Consuming moderate amounts of daily grape juice may favorably affect antioxidant defense systems and lymphocyte DNA damage in hypertensive individuals.

Combined effects of swim training and ginseng supplementation on exercise performance time, ROS, lymphocyte proliferation, and DNA damage following exhaustive exercise stress.

The purpose of this study was to analyze the combined effects of regular exercise and ginseng supplementation on peritoneal exudate ROS (reactive oxygen species), lymphocyte proliferation by splenocytes, and DNA damage following exhaustive exercise stress. Thirty-six female BALB/c mice were randomly divided into control (UT, n = 12), trained (TR, n = 12), and ginseng supplemented and trained (GT, n = 12) groups. Each group was divided into two equal subgroups where mice were studied at rest (UTre, TRre, and GTre) or immediately after exhaustive exercise stress (UTex, TRex, and GTex). Animals were bred in the animal facility, where they were housed at 22-24 degrees C and relative humidity (RH) 50-60% in a controlled environment with a 12-hour photoperiod, and provided food and water ad libitum. The trained mice underwent 10 weeks of endurance swim training (5 times/week) in water at 27-30 degrees C for 60 minutes. The analytical items examined were weight, proliferative activity, the production of ROS from peritoneal exudate cells, and DNA damage following exhaustive exercise stress (2 h exercise stress). Significant level was set at p < 0.05. The results obtained showed that the trained group had a significantly lower mean body weight than the untrained group (p < 0.05). However, there was no significant difference between UT and GT. Swim training increased swim survival time in TRex and GTex, and TRex showed the highest swim survival time. With regard to mitogenic activities of splenocytes in response to exhaustive exercise stress, all groups showed much lower lymphocyte proliferative activity when stimulated with media (Med), concanavalin A (ConA), or lipopolysaccharide (LPS) after exhaustive exercise stress. However, GTex had a higher proliferative activity than the other groups. Trained and ginseng-supplemented groups showed lower peritoneal ROS responses and lymphocyte DNA damage levels after exhaustive exercise. These findings suggest that the combined effect of swim training and ginseng supplementation sustain lymphocyte function in the presence of reduced ROS production and DNA damage following acute exercise stress.

Soy isoflavone supplementation alleviates oxidative stress and improves systolic blood pressure in male spontaneously hypertensive rats.

The aim of this study was to investigate the protective effect of isoflavone against hypertension, via the mitigation of oxidative stress and prevention of nitric oxide (NO, a potent vasodilator) reduction, in spontaneously hypertensive rats (SHR). The 8 wk-old male SHR were divided into two groups, and fed a casein-based high fat diet (120 g fat, 1 g cholesterol/kg diet) for 30 d, either with or without 10 g of soy powder (containing 31.2% of isoflavones)/kg. During the 30-d study period, tail systolic blood pressures (BP) in the control SHR group increased, from 162.4 +/- 2.3 to 177.9 +/- 5.4 mmHg (p<0.05), while the isoflavone-supplemented group benefited from a clear antihypertensive effect (160.1 +/- 1.8 to 160.2 +/- 4.9 mmHg). The serum NO and total radical trapping antioxidant potential (TRAP) were elevated in the isoflavone group. The isoflavone group also experienced a significant decrease in oxidative DNA damage in leukocytes, using comet assay. DNA damage correlated positively with incremental BP during the study, and systolic BP at the end of the study (p<0.01). Our results indicate that soy isoflavone has an antihypertensive effect, possibly through the amelioration of oxidative stress, and the augmentation of NO production, in SHR.

Concord grape juice supplementation reduces blood pressure in Korean hypertensive men: double-blind, placebo controlled intervention trial.

Many of the flavonoids found in grapes and grape products such as juice or wine have been known to exert antioxidant, anti-inflammatory, platelet inhibitory and arterial relaxing effects either in vitro, in animal studies and in human trials. This study was designed to test the effect of Concord grape juice consumption on altering blood pressure in hypertensive patients. Forty subjects were given 5.5 ml/kg body weight/day of either Concord grape juice (CGJ) or a calorie-matched placebo drink every day for 8 weeks. Blood pressure (BP) was measured on weeks 0, 4 and 8. Compared to baseline, in the CGJ group systolic BP was reduced on average by 7.2 mm Hg (p = 0.005) and diastolic BP was reduced on average by 6.2 mm Hg (p = 0.001) at the end of 8 weeks. Comparable changes in the group getting the placebo product were -3.5 mm Hg (NS) and -3.2 mm Hg (p = 0.05) Consuming Concord grape juice, which is high in polyphenolic compounds, may favorably affect BP in hypertensive individuals.