Wine, alcohol and atherosclerosis: clinical evidences and mechanisms

Atherosclerosis is a chronic inflammatory disease which may cause obstructions of the coronary, cerebral and peripheral arteries. It is typically multifactorial, most often dependent on risk factors such as hypercholesterolemia, diabetes, smoking, hypertension, sedentarism, and obesity. It is the single main cause of death in most developed countries due to myocardial infarction, angina, sudden death, and heart failure. Several epidemiological studies suggest that moderate alcohol intake, especially red wine, decrease cardiac mortality due to atherosclerosis. The alcohol effect is described by a J curve, suggesting that moderate drinkers may benefit while abstainers and heavy drinkers are at higher risk. Experimental studies indicate that most beneficial effects of drinking are attributable to flavonoids that are present in red wine, purple grape juice and several fruits and vegetables. The mechanisms include antiplatelet actions, increases in high-density lipoprotein, antioxidation, reduced endothelin-1 production, and increased endothelial nitric oxide synthase expression which causes augmented nitric oxide production by endothelial cells. These findings lead to the concept that moderate red wine drinking, in the absence of contraindications, may be beneficial to patients who are at risk of atherosclerotic cardiovascular events. Moreover, a diet based on fruits and vegetables containing flavonoids may be even more beneficial.

The action of red wine and purple grape juice on vascular reactivity is independent of plasma lipids in hypercholesterolemic patients

Although red wine (RW) reduces cardiovascular risk, the mechanisms underlying the effect have not been identified. Correction of endothelial dysfunction by RW flavonoids could be one mechanism. We measured brachial artery reactivity by high-resolution ultrasonography, plasma lipids, glucose, adhesion molecules (ICAM-1 and VCAM), and platelet function in 16 hypercholesterolemic individuals (8 men and 8 women; mean age 51.6 ± 8.1 years) without other risk factors. Twenty-four normal subjects were used as controls for vascular reactivity. Subjects randomly received RW, 250 ml/day, or purple grape juice (GJ), 500 ml/day, for 14 days with an equal wash-out period. At baseline, all 16 subjects were hypercholesterolemic (mean LDL = 181.0 ± 28.7 mg/dl) but HDL, triglycerides, glucose, adhesion molecules, and platelet function were within normal limits. Brachial artery flow-mediated dilation was significantly decreased compared to controls (9.0 ± 7.1 vs 12.1 ± 4.5%; P < 0.05) and increased with both GJ (10.1 ± 7.1 before vs 16.9 ± 6.7% after: P < 0.05) and RW (10.1 ± 6.4 before vs 15.6 ± 4.6% after; P < 0.05). RW, but not GJ, also significantly increased endothelium-independent vasodilation (17.0 ± 8.6 before vs 23.0 ± 12.0% after; P < 0.01). GJ reduced ICAM-1 but not VCAM and RW had no effect on either molecule. No significant alterations were observed in plasma lipids, glucose or platelet aggregability with RW or GJ. Both RW and GJ similarly improved flow-mediated dilation, but RW also enhanced endothelium-independent vasodilation in hypercholesterolemic patients despite the increased plasma cholesterol. Thus, we conclude that GJ may protect against coronary artery disease without the additional negative effects of alcohol despite the gender.

Coronary artery plaque burden and calcium scores in healthy men adhering to long-term wine drinking or alcohol abstinence

Observational studies suggest there are clinical benefits to moderate red wine (RW) consumption. However, the effects on coronary vasculature and overall lifestyle are unclear. We investigated whether a lifestyle of regular long-term RW consumption is associated with changes in coronary plaque burden, calcium score, carotid intima/media thickness, endothelial function, and metabolic variables, compared with alcohol abstinence. Healthy volunteers were evaluated by coronary computed tomography angiography (CTA) as well as carotid and brachial artery ultrasound. Nutritional status, psychological status, and metabolic variables were assessed. The study included 101 drinkers [aged 58.9±7.3 years (means±SD)], from wine brotherhoods, and 104 abstainers, from Anglican, Evangelical and Catholic churches both in the city of São Paulo, Brazil. No significant differences in demographics were noted. Lesion prevalence per patient assessed by coronary CTA and classified as absent (0), 1-25, 26-49, and ≥50% stenosis was similar between groups. When analyzed by individual arteries, i.e., left anterior descending, circumflex, and right coronary, prevalence was also not different. On the other hand, calcium scores were higher among drinkers than abstainers (144.4±362.2 vs 122.0±370.3; P<0.01). However, drinkers reported less history of diabetes and exercised more. RW drinkers consumed 2127.9±387.7 kcal/day while abstainers consumed 1836.0±305.0 (P<0.0001). HDL cholesterol was significantly higher among drinkers compared to abstainers (46.9±10.9 vs 39.5±9.0 mg/dL; P<0.001), while fasting plasma glucose was lower (97.6±18.2 vs 118.4±29.6 mg/dL; P<0.02). Liver enzymes were normal in both groups. In conclusion, long-term wine drinkers displayed a similar plaque burden but greater calcium score than abstainers, despite a more atherogenic diet, and the mechanisms for the increased calcium scores in the former remain speculative.

Monitoring of wine aging process by electrospray ionization mass spectrometry

The characterization of wine samples by direct insertion electrospray ionization mass spectrometry (ESI-MS), without pre-treatment or chromatographic separation, in a process denominated fingerprinting, has been applied to several samples of wine produced with grapes of the Pinot noir, Merlot and Cabernet Sauvignon varieties from the state o Rio Grande do Sul, in Brazil. The ESI-MS fingerprints of the samples detected changes which occurred during the aging process in the three grape varieties. Principal Component Analysis (PCA) of the negative ion mode fingerprints was used to group the samples, pinpoint the main changes in their composition, and indicate marker ions for each group of samples.

Stability of the anthocyanins extracted from residues of the wine industry

Anthocyanins are highly important due to their antioxidant capacity. They are the most important among the phenolic compounds and one of the main natural dyes used in the food industry. In this research, residue of processed grapes was used to investigate the presence of anthocyanins, the possibility of their extraction from the residue, and their stability. The extraction solution consisted of 70 mL of ethanol 70% and 30 mL of HCl 0.1% at pH 2.0. The results found for the processed grapes residue was 26.20 mg.100 g-1. In order to evaluate stability, caffeic acid was added at 0.5:1 w/v; 0.8:1 w/v; and 1:1 w/v concentrations. Anthocyanins extract reached the greatest stability at 0.5:1 w/v concentration, with 82.47% color retention and a half-life period of 15 days. Therefore, the use of this organic acid as a stabilizer for anthocyanins is feasible.

Effect of trellising system on grape and wine composition of Syrah vines grown in the cerrado region of Minas Gerais

Based on the concept that the trellising system affects not only sunlight interception and carbon assimilation, but also the fruitzone microclimate, which has a great impact on fruit composition and consequently on wine quality, the effect of two trellising systems - Vertical Shoot Position (VSP) and modified Geneva Double Curtain (GDC) - on wine and berry composition of Syrah grapes grown in João Pinheiro, Northeast region of Minas Gerais State, Brazil was investigated. The parameters such as pH, berry size and weight, and seeds total phenolic contents were not affected by the training system. The GDC system produced fruits with the highest Brix and lowest titratable acidity. Berries from the VSP system presented lower anthocyanin concentration than those from the GDC system. Similar results were found for the total phenolic content of the skin of grape berries from the VSP system. GDC wines were characterized by high anthocyanin content and red color, resulting in wines with high color intensity. These data suggest that in the tropical region of Minas Gerais state, with high temperature and high sunlight intensity, the trellising system, which protects bunches against excessive radiation, should be chosen.

Conversion of hydroxycinnamic acids into volatile phenols in a synthetic medium and in red wine by Dekkera bruxellensis

The conversion of p-coumaric acid, ferulic acid, and caffeic acid into 4-ethylphenol, 4-ethylguaiacol and 4-ethylcatechol was studied in Dekkera bruxellensis ISA 1791 under defined conditions in a synthetic medium and in a red wine. Liquid chromatography (HPLC-DAD) was used to quantify the phenolic acids, and gas chromatography (GC) coupled to a FID detector was used to quantify volatile phenols using a novel analytical methodology that does not require sample derivatization. Identification was achieved by gas chromatography-mass detection (GC-MS). The results show that phenolic acids concentration decreases while volatile phenols concentration increases. The proportion of caffeic acid taken up by Dekkera bruxellensis is lower than that for p-coumaric or ferulic acid; therefore less 4-ethylcatechol is formed. More important, 4-ethylcathecol synthesis by Dekkera bruxellensis in wine has never been demonstrated so far. These results contribute decisively to a better understanding of the origin of the volatile phenols in wines. The accumulation of these compounds in wine is nowadays regarded as one of the key factors of quality control.

Phenolic compounds in Merlot wines from two wine regions of Rio Grande do Sul, Brazil

In Brazil, the grape and wine production takes place mainly in the state of Rio Grande do Sul, and the region "Serra" is known as the traditional wine region. In the last years, new areas have emerged, with emphasis for the Campanha region; the red wines from this region have low acidity, little color intensity, and are wines to drink while young, even when produced from grape varieties such as Merlot and Cabernet Sauvignon. The objective of this study was to evaluate the influence of different maceration types on the phenolic compounds of Merlot wines made with grapes produced in two regions of Rio Grande do Sul, Serra and Campanha, as well as to identify the key differences between the wines produced. The localization of the vineyards seems to have more influence on the wine characteristics than the maceration type. The color due copigmentation was an important aspect in the wines made with short maceration. The effect of extended maceration was different than the expected for the Campanha region wines; the extended maceration increased the extraction of tannins resulting in greater color intensity and a greater amount of anthocyanins. The pH control seems to be a key factor for the Campanha region wines.

Use of simulated annealing in standardization and optimization of the acerola wine production

In this study, seven wine samples were prepared varying the amount of pulp of acerola fruits and the sugar content using the simulated annealing technique to obtain the optimal sensory qualities and cost for the wine produced. S. cerevisiae yeast was used in the fermentation process and the sensory attributes were evaluated using a hedonic scale. Acerola wines were classified as sweet, with 11°GL of alcohol concentration and with aroma, taste, and color characteristics of the acerola fruit. The simulated annealing experiments showed that the best conditions were found at mass ratio between 1/7.5-1/6 and total soluble solids between 28.6-29.0 °Brix, from which the sensory acceptance scores of 6.9, 6.8, and 8.8 were obtained for color, aroma, and flavor, respectively, with a production cost 43-45% lower than the cost of traditional wines commercialized in Brazil.

Research on the quality of the wine grapes in corridor area of China

The corridor area of Gansu Province is one of the most important wine grape growing regions in China, and this strip of land results in a significant difference in terms of terroir between its regions. Chemical composition and antioxidant capacity of the main wine grape varieties (Vitis vinifera L.) cultivated in the corridor area of Gansu Province in northwest China were compared. Three regions (Zhangye, Wuwei, and Jiayuguan) were selected to explain the influence of soil and climate conditions on the quality of wine grapes. This study aims to investigate the effect of different regions on berry composition and antioxidant capacity, providing a general evaluation of red and white wine grapes quality in the corridor area of China. The results showed that ‘Merlot’ grapes grown in Zhangye had the best quality among the different varieties in the three regions of Gansu evaluated. The moderate temperature and nitrogen deficiency were associated with improved fruit quality. It was identified that the most suitable grape variety from Zhangye is ‘Merlot’, and that ‘Cabernet Sauvignon’ and ‘Italian Resling’ are the most suitable varieties from Wuwei and Jiayuguan, respectively.

Fresh meat quality of pigs fed diets with different fatty acid profiles and supplemented with red wine solids

Abstract Three groups of pigs were fed three different diets, namely a diet rich in saturated fatty acids (palm oil-based, PO), a polyunsaturated fatty acid (PUFA)-rich diet (corn oil-based, CO), and a PUFA-rich diet (corn oil-based) supplemented with red wine solids (RWS), which was added to the diet (CO+RWS) in order to assess the protective effect on the oxidative status of the pork meat. The addition of corn oil favourably modulates the FAs profile of the backfat, and to a lesser extent of the intramuscular fat of semimembranosus muscle, without causing adverse effects on the meat quality or on its oxidative stability. Moreover, these parameters were not affected by the addition of the RWS in the CO+RWS diet.

Timing of di-ammonium phosphate addition to fermenting chardonnay must : effect on nitrogen utilization, ethyl-carbamate formation, and CAR1 expression by yeast /

The maximum amount of ethyl carbamate (EC), a known animal carcinogen produced by the reaction of urea and ethanol, allowed in alcoholic beverages is regulated by legislation in many countries. Wine yeast produce urea by the metabolism of arginine, the predominant assimilable amino acid in must. This action is due to arginase (encoded by CARl). Regulation of CARl, and other genes in this pathway, is often attributed to a well-documented phenomenon known as nitrogen catabolite repression. The effect of the timing of di-ammonium phosphate (DAP) additions on the nitrogen utilization, regulation of CARl, and EC production was investigated. A correlation was found between the timing of DAP addition and the utilization of nitrogen. When DAP was added earlier in the fermentations, less amino nitrogen and more ammonia nitrogen was sequestered from the media by the cells. It was also seen that early DAP addition led to more total nitrogen being used, with a maximal difference of ~25% between fermentations where no DAP was added versus addition at the start of the fermentation. The effect of the timing ofDAP addition on the expression of CARJ during fermentation was analyzed via northern transfer and the relative levels of CARl expression were determined. The trends in expression can be correlated to the nitrogen data and be used to partially explain differences in EC formation between the treatments. EC was quantified at the end of fermentation by GC/MS. In Montrachet yeast, a significant positive correlation was found between the timing of DAP addition, from early to late, and the final EC concentration m the wine (r = 0.9226). In one of the fermentations, EC levels of 30.5 ppb was foimd when DAP was added at the onset of fermentation. A twofold increase (69.5 ppb) was observed when DAP was added after 75% of the sugars were metabolized. When no DAP was added, the ethyl carbamate levels are comparable at a value of 38 ppb. In contrast, the timing of DAP additions do not affect the level EC produced by the yeast ECU 18 in this manner. The study of additional yeast strains shows that the effect of DAP addition to fermentations is strain dependent. Our results reveal the potential importance of the timing of DAP addition to grape must with respect to EC production, and the regulatory effect of DAP additions on the expression of genes in the pathway for arginine metabolism in certain wine yeast strains.

The osmoadaptive response of the wine yeast Saccharomyces cerevisiae K1-V1116 during icewine fermentation

The adapted metabolic response of commercial wine yeast under prolonged exposure to concentrated solutes present in Icewine juice is not fully understood. Presently, there is no information regarding the transcriptomic changes in gene expression associated with the adaptive stress response ofwine yeast during Icewine fermentation compared to table wine fermentation. To understand how and why wine yeast respond differently at the genomic level and ultimately at the metabolic level during Icewine fermentation, the focus ofthis project was to identify and compare these differences in the wine yeast Saccharomyces cerevisiae KI-Vll16 using cDNA microarray technology during the first five days of fermentation. Significant differences in yeast gene expression patterns between fermentation conditions were correlated to differences in nutrient utilization and metabolite production. Sugar consumption, nitrogen usage and metabolite levels were measured using enzyme assays and HPLC. Also, a small subset of differentially expressed genes was verified using Northern analysis. The high osmotic stress experienced by wine yeast throughout Icewine fermentation elicited changes in cell growth and metabolism correlating to several fermentation difficulties, including reduced biomass accumulation and fermentation rate. Genes associated with carbohydrate and nitrogen transport and metabolism were expressed at lower levels in Icewine juice fermenting cells compared to dilute juice fermenting cells. Osmotic stress, not nutrient availability during Icewine fermentation appears to impede sugar and nitrogen utilization. Previous studies have established that glycerol and acetic acid production are increased in yeast during Icewine fermentation. A gene encoding for a glycerollW symporter (STL1) was found to be highly expressed up to 25-fold in the i Icewine juice condition using microarray and Northern analysis. Active glycerol transport by yeast under hyperosmotic conditions to increase cytosolic glycerol concentration may contribute to reduced cell growth observed in the Icewine juice condition. Additionally, genes encoding for two acetyl CoA synthetase isoforms (ACSl and ACS2) were found to be highly expressed, 19- and II-fold respectively, in dilute juice fermenting cells relative to the Icewine juice condition. Therefore, decreased conversion of acetate to acetyl-CoA may contribute to increased acetic acid production during Icewine fermentation. These results further help to explain the response of wine yeast as they adapt to Icewine juice fermentation. ii

The effect of pH and ethanol on the astringent sub-qualities of red wine, and the implications for optimum grape maturity /

The first objective of this study was to identify appropriate sensory descriptors to assess the astringent sub-qualities of red wine. The influence of pH and ethanol on the sensation of astringency in red wine was evaluated, using a de-alcoholized red wine. A portion of the wine was adjusted to the pH values of 3.2, 3.4, 3.6 and 3.8, and another portion was adjusted to ethanol concentrations of 0%, 6%, 12%, and 15%. In addition, the pH 3.4 and 3.6 treatments were adjusted to an ethanol concentration of 12% and 15% all wines were then assessed sensorially and seventeen terms were identified, through panel discussion, to describe the mouth-feel and taste qualities: velvet, aggressive, silk/satin, dry, fleshy, unripe, pucker viscosity, abrasive, heat, chewy, acidity, grippy/adhesive, bitter, balance, overall astringency, and mouth-coat. Descriptive analysis profiling techniques were used to train the panel and measure the intensity of these attributes. It was found that decreasing pH values (averaged across all ethanol concentrations) showed an increase in the overall astringency of the wine. The combined treatments of ethanol and pH, real wine parameters (pH 3.4 and 3.6; 12% and 15% ethanol) did not have an effect on the perception of the astringent sub-qualities of the wine. A time intensity study was also included using the pH and ethanol adjusted wines, which showed that as the ethanol level of the wines increased so did the time to maximum intensity. The second objective was to identify appropriate sensory descriptors to evaluate the influence of grape maturity and maceration technique (grape skin contact) on the astringency sub-qualities of red vinifera wines from Niagara. The grapes were harvested across two dates, representing an early harvest and a late harvest. A portion of the Cabernet Sauvignon grapes wine was divided into three maceration treatments of oneweek maceration, standard two-week maceration, three-week maceration, and MCM. Another portion of both the early and late harvest Cabernet Sauvignon grapes were chaptalized to yield a final ethanol concentration of 14.5%. The wines were assessed sensorially and thirteen terms were identified, through panel discussion, to describe the mouth-feel and taste qualities: carbon dioxide, pucker, acidity, silk/chamois, dusty/chalky/powdery, sandpaper, numbing, grippy/adhesive, dry, mouthcoat, bitter, balance and, overall astringency. Descriptive analysis techniques were used to train the panel and measure the intensity of these attributes. The data revealed few significant differences in the mouth-feel of the wines with respect to maturity; which included differences in overall astringency and balance. There were varietal differences between Cabernet Sauvignon, Cabernet Franc, and Pinot Noir and differences for Cabernet Sauvignon wines due to the length and manner of maceration and as a result of chaptalization. Statistical analysis revealed a more complex mouth-feel for the Pinot Noir wines; and an increase in the intensity of the astringent sub-qualities as a result of the addition of sugar to the wines. These findings have implications for how processing decisions, such as optimum grape maturity and vinification methods may affect red wine quality.

Investigations of 2-alkyl-3-methoxypyrazines in three ladybug species and wine /

Investigations of 2-alkyl-3-methoxypyrazines (2-isopropyl-3-methoxypyra2ine, 2- secbutyl-3-methoxypyrazine and 2-isobutyl-3-niethoxypyrazine) in ladybug species {Coleoptera: Coccinellidae) and wine samples have been conducted. Headspace sampling coupled with gas chromatography-mass spectrometry was used to determine amounts of 2-alkyl-3-methoxypyra2ines in the ladybug species. Hippodamia convergens had the highest amount of alkybnethoxypyrazines, followed by Harmoma axyridis and the least in Coccinella septempunctata. Using a solvent extraction method, the precoccinelline alkaloid was found present in Hippodamia convergens and Coccinella septempunctata but not Harmonia axyridis. Steam distillation followed by a soHd phase extraction method as a sample preparation technique, enhanced detection while the isotope dilution method afforded accurate quantitation of the alkyknethoxypyrazines in the wine samples. Both ladybug-tainted and commercial wine samples were found to contain the 2- alkyl-3-methoxypyrazines. Wine samples prepared in 2001 generally contained higher levels than the corresponding 2003 samples. Levels of the 2-alkyl-3-methoxypyrazines found in the commercial wines ranged from a minimum value of 6 ng/L to 260 ±10 ng/L. Analyses revealed that for both ladybug species and wine samples, the 2- isopropyl-3-methoxypyrazine had the highest concentration, followed by 2-isobutyl- 3-methoxypyrazine and the least being the 2-secbutyl-3-methoxypyrazine. Possible contamination of the wine samples by ladybugs is thoroughly discussed. Furthermore, attempts to remove or reduce the levels of the alkylmethoxypyrazines with molecularly imprinted polymers from wine samples are presented in detail.