Effect of simultaneous coffee/caffeine and ethanol administration on sperm quality and reproductive hormones: an experimental study in Sprague Dawley rats.

Chronic alcohol or coffee/caffeine consumption has been associated with negative effects on male reproductive system. The present study evaluated the possible interaction (combination treatment) of coffee/caffeine and alcohol and its resultant consequence on sperm parameters in rats. Ten groups of adult Sprague Dawley rats (n = 5 per group) were orally gavaged with different quantities of coffee (≈5, 10, and 20 mg/kg caffeine); 30% v/v ethanol (0.5, 1, and 2 g/kg); coffee + ethanol or vehicle (distilled water) daily for 60 days. Sperm parameters, serum sex hormone levels, together with antioxidants and lipid peroxidation status of testis homogenate were evaluated. Single and combined administration of coffee/caffeine and ethanol caused a reduction in sperm count dose-dependently (p = 0.0002), but this effect was highest in the combined treatment. Sperm viability, motility, and morphology were unaffected by coffee or ethanol administration, but the combination produced detrimental effects on them. While changes in follicle stimulating hormone were not significant, testosterone and luteinizing hormone levels were decreased significantly and dose-dependently in all experimental groups (p < 0.05-0.0001). This effect was most pronounced in the groups that received the combination. Testicular superoxide dismutase and catalase activities, and reduced glutathione and malondiadehye (MDA) levels were not affected by coffee/caffeine, but ethanol decreased (p < 0.05) the antioxidants and elevated MDA levels mostly at 2 g/kg. The combination reduced (p < 0.0001) the antioxidants and increased MDA levels in testis homogenates at all doses. Combination of coffee/caffeine and ethanol potentiates their detrimental effects on sperm quality and reproductive hormones via reduction in testicular antioxidants activities and elevation of lipid peroxidation in rats.

El consumo de café: su asociación con el riesgo de padecer cáncer / Coffee consumption: its association with the risk of cancer

RESUMEN El café y su impacto en la salud es un tema en el que resulta válido profundizar. Históricamente, el consumo de café se ha asociado con efectos adversos, como problemas cardiovasculares y varios tipos de cáncer. Pero en gran cantidad de fuentes bibliográficas contemporáneas se enfatiza en los efectos beneficiosos de su consumo, sin mencionar los daños que puede ocasionar a la salud. Se hace esta revisión bibliográfica con el objetivo de profundizar en lo más actualizado sobre los beneficios y perjuicios del consumo del café y su relación con la aparición del cáncer. En la revisión se consultaron artículos de las bases de datos PubMed, SciELO, ClinicalKey y LILACS. Se constató que el consumo de café no se asocia con la aparición de diferentes tipos de cánceres, y que el consumo moderado aporta propiedades protectoras para la salud. Teniendo en cuenta el carácter multifactorial del cáncer, los autores consideran que suponer que el consumo de esta bebida puede impedir carcinogénesis, es una tesis que debe ser interpretada con cautela (AU).

ABSTRACT Coffee and its impact on health is a topic on which it is valid to deepen. Historically, coffee consumption has been associated with side effects, such as cardiovascular problems and several types of cancer. But many contemporary bibliographic sources emphasize the beneficial effects of its consumption, without mentioning the damage it can cause to health. This bibliographic review is done with the aim of deepening into the most updated knowledge about the benefits and harms of coffee consumption and its relationship with the appearance of cancer. Articles from PubMed, SciELO, ClinicalKey and LILACS databases were reviewed. It was found that coffee consumption is not associated with the appearance of different types of cancers, and that moderate consumption provides protective properties for health. In view of the multifactorial character of cancer, the authors consider that assuming that the consumption of this drink can prevent carcinogenesis is a thesis that should be taken with caution (AU).

Chemical Analysis, Toxicity Study, and Free-Radical Scavenging and Iron-Binding Assays Involving Coffee (Coffea arabica) Extracts.

We aimed to analyze the chemical compositions in Arabica coffee bean extracts, assess the relevant antioxidant and iron-chelating activities in coffee extracts and instant coffee, and evaluate the toxicity in roasted coffee. Coffee beans were extracted using boiling, drip-filtered and espresso brewing methods. Certain phenolics were investigated including trigonelline, caffeic acid and their derivatives, gallic acid, epicatechin, chlorogenic acid (CGA) and their derivatives, p-coumaroylquinic acid, p-coumaroyl glucoside, the rutin and syringic acid that exist in green and roasted coffee extracts, along with dimethoxycinnamic acid, caffeoylarbutin and cymaroside that may be present in green coffee bean extracts. Different phytochemicals were also detected in all of the coffee extracts. Roasted coffee extracts and instant coffees exhibited free-radical scavenging properties in a dose-dependent manner, for which drip coffee was observed to be the most effective (p < 0.05). All coffee extracts, instant coffee varieties and CGA could effectively bind ferric ion in a concentration-dependent manner resulting in an iron-bound complex. Roasted coffee extracts were neither toxic to normal mononuclear cells nor breast cancer cells. The findings indicate that phenolics, particularly CGA, could effectively contribute to the iron-chelating and free-radical scavenging properties observed in coffee brews. Thus, coffee may possess high pharmacological value and could be utilized as a health beverage.

Risk assessment of coffees of different qualities and degrees of roasting.

During the coffee beans roasting process, occurs the formation of polycyclic aromatic hydrocarbons, which are associated with the incidence of cancer in humans. This study aimed to evaluate the influence of coffee bean quality and roasting degree regarding mutagenicity, cytotoxicity and genotoxicity. Six samples of coffee drink made with roasted and ground Coffea arabica beans from different qualities and roast degrees were used after freeze-drying. Both commercial and special quality grains suffered light, medium and dark roasting. According to the Salmonella/microsome assay, the highest concentration of commercial grain sample (dark roast) significantly increased the number of revertants of the TA98 strain in the absence of metabolization. All the samples induced cytotoxicity to HepG2 cells. These effects can be ranked in the following order from most to least toxic: medium roast - special grain > light roast - special grain > dark roast - commercial grain > dark roast - special grain > light roast - commercial grain > medium roast - commercial grain. None of the samples induced genotoxicity in HepG2 cells. Our findings show that the harmful effects of coffee depend not only on the degree of roasting but also on the grain quality.

Exploration of the cellular effects of the high-dose, long-term exposure to coffee roasting product furan and its by-product cis-2-butene-1,4-dial on human and rat hepatocytes.

Coffee is the most popular hot beverage and caffeine is the most used psychoactive drug in the world. Roasting of coffee beans leads to the generation of minute quantities of undesirable compounds, such as furan. It is now thought that the toxicity of furan derives from its processing by CYP450 family of detoxifying enzymes, leading to the formation of cis-2-butene-1,4-dial (BDA). BDA has known cytotoxicity capacities, binding to proteins, nucleic acids, and glutathione (GSH). BDA also appears to mediate furan's toxic effects, since the inhibition of CYP450 family impedes the aforementioned toxicological effects of furan. There are some studies performed on furan's toxicity, but very few on BDA. Furthermore, the doses used in these studies appear to be fairly high when compared with the expected dosage one could be exposed to in a standard day. As such, to understand if furan and BDA could have toxic effects using more realistic doses and longer time frames, human and rat hepatocytes were exposed to furan or BDA for up to 96 h, and several biochemical parameters were assessed. We report here that human hepatocytes were more sensitive than rat's, in particular to furan, for we show a decrease in MTT reduction, ATP levels and increase in carbonyl formation and 8-OHdG accumulation in the longer time points. BDA was mostly ineffective, which we attribute to a low import rate into the cells. In conclusion, we show that there is potential for harm from furan in high doses, which should be carefully addressed.

Effects of Coffee Extracts with Different Roasting Degrees on Antioxidant and Anti-Inflammatory Systems in Mice.

Coffee roasting affects the taste, color, and aroma of coffee. The Maillard reaction, a major reaction during the roasting process, produces melanoidin, which affects the overall antioxidant capacity and anti-inflammatory effects of coffee. In this experiment, coffee roasting was divided into four degrees: Light, Medium, City, and French. To examine the in vivo antioxidant and anti-inflammatory effects of coffee extracts with different roasting degrees, we used 10-week-old male C57BL/6 mice. Mice were pre-treated with coffee extracts for 10 days by oral gavage (300 mg/Kg.B.W). After the last pre-treatment, lipopolysaccharide (LPS, 15 mg/Kg.B.W) was injected intraperitoneally for immune stimulation. Histopathological analysis showed that hepatic portal vein invasion and liver necrosis were severe in the LPS-treated group. However, these phenomena were greatly ameliorated when mice were pre-treated with Light- or Medium-roasted coffee extracts. Hepatic glutathione level was increased in the French group but decreased in the LPS-stimulated group. When mice were treated with LPS, mRNA expression level of tumor necrosis factor-alpha (TNF-α) was increased, whereas TNF-α expression was significantly reduced in the Light and Medium groups. Treatment with coffee extracts decreased the mRNA expression levels of interleukin 6 (IL-6) in mice stimulated by LPS, regardless of coffee roasting degrees. These effects decreased with the increasing coffee roasting degree. Results of luciferase reporter assay revealed that these effects of coffee extracts were transcriptionally regulated by the NF-κB pathway. Taken together, these results suggest that the roasting degree affects the antioxidant and anti-inflammatory effects of coffee extracts.

Roasted and green coffee extracts show antioxidant and cytotoxic activity in myoblast and endothelial cell lines in a cell specific manner.

Coffee is one of the most highly consumed beverages with potential beneficial health implications, however its molecular mechanism of action has not been completely elucidated yet. To that cause, the polyphenolic composition of different coffee extracts (from Light, Medium and Dark roasts as well as green beans) was examined by UHPLC-HRMS analysis, indicating chlorogenic acids isomers as the main constituents. In the following step, the toxicity of the extracts was tested in myoblasts and endothelial cells and differential toxicity of green and roasted samples was displayed as the myoblasts were more sensitive to green coffee extracts, in contrast to the endothelial cells. Subsequently, biologically relevant, non-cytotoxic extract concentrations were administered to explore their potential effect on cell redox status using flow cytometry and spectrophotometric assays. The results indicated that all coffee extracts improved cell redox status, however differences were observed between the two different cell lines tested, implying that coffee compounds display cell- and tissue-specificity. Glutathione levels were increased in almost all cases up to 70%, while the roasting degree affected the free radical scavenging potential of the extracts and their ability to protect from macromolecular oxidation as exhibited by the differences in ROS, CARB and TBARS levels, especially in the myoblasts.

Cells Deficient in the Fanconi Anemia Protein FANCD2 are Hypersensitive to the Cytotoxicity and DNA Damage Induced by Coffee and Caffeic Acid.

Epidemiological studies have found a positive association between coffee consumption and a lower risk of cardiovascular disorders, some cancers, diabetes, Parkinson and Alzheimer disease. Coffee consumption, however, has also been linked to an increased risk of developing some types of cancer, including bladder cancer in adults and leukemia in children of mothers who drink coffee during pregnancy. Since cancer is driven by the accumulation of DNA alterations, the ability of the coffee constituent caffeic acid to induce DNA damage in cells may play a role in the carcinogenic potential of this beverage. This carcinogenic potential may be exacerbated in cells with DNA repair defects. People with the genetic disease Fanconi Anemia have DNA repair deficiencies and are predisposed to several cancers, particularly acute myeloid leukemia. Defects in the DNA repair protein Fanconi Anemia D2 (FANCD2) also play an important role in the development of a variety of cancers (e.g., bladder cancer) in people without this genetic disease. This communication shows that cells deficient in FANCD2 are hypersensitive to the cytotoxicity (clonogenic assay) and DNA damage (γ-H2AX and 53BP1 focus assay) induced by caffeic acid and by a commercial lyophilized coffee extract. These data suggest that people with Fanconi Anemia, or healthy people who develop sporadic mutations in FANCD2, may be hypersensitive to the carcinogenic activity of coffee.

Coffee drinking and colorectal cancer risk: an evaluation based on a systematic review and meta-analysis among the Japanese population.

OBJECTIVE: It remains unclear whether coffee drinking is associated with colorectal cancer risk. We performed a systematic review and meta-analysis of epidemiologic studies on this issue among the Japanese population. METHODS: Original data were obtained from MEDLINE searches using PubMed or from searches of the 'Ichushi' database, complemented with manual searches. Meta-analysis was performed by using the random effects model to estimate the summary relative risk with 95% confidence interval according to the study design. The final judgment was made based on a consensus of the research group members with consideration for both epidemiological evidence and biological plausibility. RESULTS: We identified five cohort studies and nine case-control studies. Of these, one cohort study reported a strong inverse association (in women only), whereas three case-control studies reported a strong inverse association with colon or rectal cancer. In meta-analysis, high consumption of coffee was not appreciably associated with colorectal cancer risk among cohort studies, whereas it was associated with significantly lower risk of colorectal or colon cancer among case-control studies. The summary relative risk/odds ratio (95% confidence interval) for the highest versus lowest categories of coffee consumption was 0.95 (0.77-1.17) and 0.78 (0.65-0.95) for cohort and case-control studies, respectively. CONCLUSIONS: The evidence is insufficient to support that coffee drinking increases or decreases the risk of colorectal cancer among the Japanese population.

Analysis of mycotoxins in coffee and risk assessment in Spanish adolescents and adults.

Mycotoxins are toxic compounds produced by fungal secondary metabolism that cause toxicological effects. Coffee is a highly popular beverage that is susceptible to contamination by mycotoxigenic fungi. The aim of the present study was to determine the presence of the following 21 mycotoxins in coffee using liquid chromatography tandem mass spectrometry (LC-MS/MS-IT): aflatoxin B1, B2, G1 and G2; ochratoxin A; nivalenol; deoxynivalenol; 3-acetyldeoxynivalenol; 15-acetyldeoxynivalenol; diacetoxyscirpenol; neosolaniol; T-2 and HT-2 toxin; sterigmatocystin; enniatin A, A1, B, and B1; beauvericin; and fumonisin B1 and B2. We aimed to determine differences by coffee process (coffee maker, electrical machine, soluble and traditional Turkish process) and to calculate the estimated daily intake (EDI) and risk assessment of mycotoxins from coffee consumption using deterministic approach at various scenarios of food consumption in Spanish adolescents and adults. The results demonstrate that all studied mycotoxins were detected in samples with mean concentrations ranging from 0.69 µg/kg to 282.89 µg/kg. Eleven percent of samples did not show contamination with legislated mycotoxins. Only 15-acetyldeoxynivalenol, deoxynivalenol, neosolaniol, fumonisin B1, and ochratoxin A exhibited significant differences between methods of coffee brewing. The results show that coffee intake does not represent a potential risk for consumers with respect to individual mycotoxin contamination.

Furan in heat-treated foods: formation, exposure, toxicity, and aspects of risk assessment.

Furan is formed in a variety of heat-treated foods through thermal degradation of natural food constituents. Relatively high levels of furan contamination are found in ground roasted coffee, instant coffee, and processed baby foods. European exposure estimates suggest that mean dietary exposure to furan may be as high as 1.23 and 1.01 µg/kg bw/day for adults and 3- to 12-month-old infants, respectively. Furan is a potent hepatotoxin and hepatocarcinogen in rodents, causing hepatocellular adenomas and carcinomas in rats and mice, and high incidences of cholangiocarcinomas in rats at doses ≥ 2 mg/kg bw. There is therefore a relatively low margin of exposure between estimated human exposure and doses that cause a high tumor incidence in rodents. Since a genotoxic mode of action cannot be excluded for furan-induced tumor formation, the present exposures may indicate a risk to human health and need for mitigation. This review summarizes the current knowledge on mechanisms of furan formation in food, human dietary exposure to furan, and furan toxicity, and highlights the need to establish the risk resulting from the genotoxic and carcinogenic properties of furan at doses lower than 2 mg/kg bw.

Safety studies on products from whole coffee fruit.

The fruit of the coffee plant, Coffea arabica, has high phenolic antioxidant and phytonutrient content and could be a beneficial food ingredient. However, the fruit has historically been discarded for the favored harvesting of the coffee bean alone. CoffeeBerry products are derived from the whole fruit and include a ground whole powder, a water extract, and a more recently developed water-ethanol extract. The safety of CoffeeBerry products was evaluated in three genotoxicity studies, three short-term oral toxicity studies, and a 90-day dietary toxicity study. Bacterial mutagenicity studies and a micronucleus test using murine peripheral cells demonstrated that none of the three products showed mutagenic or genotoxic potential. In the short-term studies, despite palatability issues, female rats showed a tolerance for whole powder and ethanol extract at doses up to 8800 mg/kg bw/day. Male rats also exhibited palatability issues and tolerated lower doses of approximately 4000 mg/kg bw/day ethanol extract via gavage and approximately 2100 mg/kg bw/day whole powder or water extract in the diet. When fed in the diet to Sprague-Dawley rats for 90 days, ethanol extract showed no adverse effects at dietary concentrations of up to 5% (approximately 3446 and 4087 mg/kg bw/day for male and female rats, respectively).

Coffee consumption, genetic susceptibility and bladder cancer risk.

OBJECTIVE: We evaluated the bladder cancer risk associated with coffee consumption in a case-control study in Spain and examined the gene-environment interactions for genetic variants of caffeine-metabolizing enzymes. METHODS: The analyses included 1,136 incident cases with urothelial carcinoma of the urinary bladder and 1,138 controls. Odds ratios (OR) and 95% confidence intervals (CI) were adjusted for area, age, gender, amount of cigarette smoking, and years since quitting among former smokers. RESULTS: The OR (95% CI) for ever consumed coffee was 1.25 (0.95-1.64). For consumers of 1, 2, 3, and 4 or more cups/day relative to never drinkers, OR were, respectively, 1.24 (0.92-1.66), 1.11 (95% CI 0.82-1.51), 1.57 (1.13-2.19), and 1.27 (0.88-1.81). Coffee consumption was higher in smokers compared to never smokers. The OR for drinking at least 4 cups/day was 1.13 (0.61-2.09) in current smokers, 1.57 (0.86-2.90) in former smokers, and 1.23 (0.55-2.76) in never smokers. Gene-coffee interactions evaluated in NAT2, CYP1A2, and CYP2E1-02 and CYP1A1 were not identified after adjusting for multiple testing. CONCLUSION: We observed a modest increased bladder cancer risk among coffee drinkers that may, in part, be explained by residual confounding by smoking. The findings from the gene-coffee interactions need replication in further studies.

Coffee consumption and risk of type 2 diabetes, cardiovascular diseases, and cancer.

Numerous epidemiological studies have evaluated the association between coffee consumption and risk of type 2 diabetes, coronary heart disease, and various cancers. This paper briefly reviews the evidence for a relation between coffee consumption and these conditions, with particular attention to methodological issues. Several early studies suggested that coffee consumption could result in a marked increase in risk of coronary heart disease and several types of cancer. However, more recent prospective cohort studies that are less prone to selection and information bias have not confirmed these findings. High consumption of unfiltered types of coffee, such as French press and boiled coffee, has been shown to increase low-density-lipoprotein-cholesterol concentrations. In addition, limiting caffeinated coffee intake during pregnancy seems a prudent choice. However, evidence has been accumulating that frequent consumption of coffee may reduce risk of type 2 diabetes and liver cancer. Further experimental studies are warranted to elucidate the underlying mechanisms and possibly identify the components in coffee that are responsible for these putative effects. In sum, the currently available evidence on coffee and risk of cardiovascular diseases and cancer is largely reassuring, and suggests that, for the general population, addressing other health-related behaviors has priority for the prevention of chronic diseases.

Categorizing mistaken false positives in regulation of human and environmental health.

One of the concerns often voiced by critics of the precautionary principle is that a widespread regulatory application of the principle will lead to a large number of false positives (i.e., over-regulation of minor risks and regulation of nonexisting risks). The present article proposes a general definition of a regulatory false positive, and seeks to identify case studies that can be considered authentic regulatory false positives. Through a comprehensive review of the science policy literature for proclaimed false positives and interviews with authorities on regulation and the precautionary principle we identified 88 cases. Following a detailed analysis of these cases, we found that few of the cases mentioned in the literature can be considered to be authentic false positives. As a result, we have developed a number of different categories for these cases of "mistaken false positives," including: real risks, "The jury is still out," nonregulated proclaimed risks, "Too narrow a definition of risk," and risk-risk tradeoffs. These categories are defined and examples are presented in order to illustrate their key characteristics. On the basis of our analysis, we were able to identify only four cases that could be defined as regulatory false positives in the light of today's knowledge and recognized uncertainty: the Southern Corn Leaf Blight, the Swine Flu, Saccharin, and Food Irradiation in relation to consumer health. We conclude that concerns about false positives do not represent a reasonable argument against future application of the precautionary principle.

La acrilamida, contaminante químico de procesado: revisión / Acrylamide formation in foods: a review Acrylamide

La acrilamida, 'probable carcinógeno para los humanos',mutágeno de categoría 2 y tóxico para la reproducción de categoría 3según la UE, se comporta como neurotóxico tras exposiciones agudas. A pesar de que se recomienda disminuir los niveles de exposición, el tabaquismo, la exposición ocupacional y la exposición dietética son fuentes de acrilamida para el hombre. De entre todos los alimentos, son los ricos en carbohidratos y los elaborados a altas temperaturas, los que mayores niveles de este tóxico presentan. En la presente revisión se explica la formación de acrilamida en los alimentos, se describen sus efectos tóxicos, se citan los métodos analíticos usados en su determinación, se recopilan datos sobre los niveles detectados en distintos alimentos y se enumeran los datos más recientes sobre la ingesta en distintas poblaciones (AU)

Acrylamide formation in foods: A review Acrylamide,'probable carcinogenic for humans', mutagenic type 2 and toxic for reproduction type 3 for the European Union, produces neurotoxicity after acute expositions. Although recommendations are given to minimize the exposition levels to this compound, smoking and occupational and dietary exposures are important acrylamide sources. Among foods, those rich in carbohydrates and those cooked at high temperatures present the highest acrylamide concentrations. The present revision points out the acrylamide formation in foods, its toxic effects and the analytical methods used in its determination. Moreover, data are given about acrylamide levels in foods and dietary intakes in different populations (AU)

Coffee consumption and stomach cancer risk in a cohort of Swedish women.

Few prospective studies have examined the relationship between coffee consumption and risk of stomach cancer, and the findings have been inconsistent. We prospectively investigated the association of long-term coffee consumption with risk of stomach cancer in a population-based cohort study of 61,433 Swedish women. Information on coffee consumption was collected with a food-frequency questionnaire at baseline (1987-1990) and updated in 1997. During a mean follow-up of 15.7 years from 1987 through June 2005, 160 incident cases of stomach cancer were diagnosed. Coffee consumption was positively associated with the risk of stomach cancer. Compared to women who consumed 1 or fewer cups of coffee per day, the multivariate hazard ratios were 1.49 (95% = 0.97-2.27) for women who drank 2-3 cups per day and 1.86 (95% CI = 1.07-3.25) for those who drank 4 or more cups per day (p for trend = 0.01). An increase of 1 cup of coffee per day was associated with a statistically significant 22% increased risk of stomach cancer (hazard ratio = 1.22; 95% CI = 1.05-1.42). These prospective data suggest that coffee consumption may increase the risk of stomach cancer in a dose-response manner. This finding needs to be confirmed in other prospective studies.

Coffee increases levels of urinary 8-hydroxydeoxyguanosine in rats.

We examined whether coffee or chlorogenic acid inhibits 8-hydroxydeoxyguanosine (8-OHdG), one of the major forms of oxidative DNA damage, in vivo and in vitro. Forty-eight male Wistar rats were assigned to three treatment groups: a control-diet group (n=16; coffee-free diet), a 0.62% coffee-diet group (n=16, dose of coffee consumed 125 mg/day), and a 1.36% coffee-diet group (n=16, dose of coffee consumed 275 mg/day) and were maintained on an experimental diet for 130 days. The coffee-diet resulted in significantly increased excretion of urinary chlorogenic acid, with the 0.62 and 1.36% coffee-diets resulting in 14.00+/-0.94 and 15.80+/-0.41 ng/mg creatinine, respectively, whereas in control rats it was not detected. Using monoclonal antibody to measure 8-OHdG, it was revealed that coffee led to a significant increase in excretion of urinary 8-OHdG on day 130 (46.62+/-13.42 ng/mg creatinine in 0.62% coffee-diet group and 64.58+/-20.15 ng/mg creatinine in 1.36% coffee-diet group, P<0.05 vs. control; control group 10.89+/-2.59 ng/mg creatinine). Furthermore, to clarify the mechanism of 8-OHdG formation by coffee, we investigated the in vitro effect of chlorogenic acid on 8-OHdG formation in human placental DNA. Chlorogenic acid alone did not lead to an increase of 8-OHdG formation, but dramatically increased it in the presence of cupric chloride and H(2)O(2). However, chlorogenic acid and cupric chloride decreased the formation of 8-OHdG in the presence of H(2)O(2). Based on these results, a possible mechanism of 8-OHdG formation in vivo by chlorogenic acid is discussed.