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PubMed2005

Rejuvenation of antioxidant system in central nervous system of aged rats by grape seed extract.

Oxidative stress is considered as a major risk factor that contributes to age-related increase in lipid peroxidation and declined antioxidants in the central nervous system during aging. Grape seed extract, one of the bioflavonoid, is widely used for its medicinal properties. In the present study, we evaluated the role of grape seed extract on lipid peroxidation and antioxidant status in discrete regions of the central nervous system of young and aged rats. Male albino rats of Wistar strain were divided into four groups: Group I-control young rats, Group II-young rats treated with grape seed extract (100 mg/kg body weight) for 30 days, Group III-aged control rats and Group IV-aged rats supplemented with grape seed extract (100 mg/kg body weight) for 30 days. Age-associated increase in lipid peroxidation was observed in the spinal cord, cerebral cortex, striatum and the hippocampus regions of aged rats (Group III). Activities of antioxidant enzymes like superoxide dismutase, catalase, glutathione peroxidase and levels of non-enzymic antioxidants like reduced glutathione, Vitamin C and Vitamin E were found to be significantly decreased in all the brain regions studied in aged rats when compared to young rats. However, normalized lipid peroxidation and antioxidant defenses were reported in the grape seed extract-supplemented aged rats. These findings demonstrated that grape seed extract enhanced the antioxidant status and decreased the incidence of free radical-induced lipid peroxidation in the central nervous system of aged rats.

Aging↗
PubMed2006

Muscadine grape products intake, diet and blood constituents of non-diabetic and type 2 diabetic subjects.

OBJECTIVE: Red wines and grape juices contain polyphenolics with antioxidant and antiplatelet properties that may be protective against oxidative stress leading to hypertension, insulin resistance, and type 2 diabetes (T2D). This study evaluated the effects of supplementing meals of subjects with 150 mL of muscadine grape juice (MJ), muscadine grape wine (MW), and dealcoholized muscadine grape wine (Dz-W) on glycemic indices, blood constituents, lipid profile, anthropometric, and nutrient intakes of healthy and T2D subjects over a 28-d period. METHODS: Subjects with T2D were assigned to take MJ, MW, or Dz-W. Non-diabetics consumed MJ and controls were given no test drinks. Several metabolic indicators associated with diabetic conditions were measured at baseline and repeated after 28 d. RESULTS: Diabetics given MW and Dz-W showed lower levels of blood glucose, insulin, and glycated hemoglobin, indicating better glycemic control. Elevated dietary vitamin C and E levels were observed in diabetics given Dz-W, indicating improved antioxidant status. Decreased red blood cell membrane saturated fatty acids and increased mono- and polyunsaturated fatty acids for subjects with T2D given MW suggested improved membrane fluidity. Lower sodium and chloride values for subjects T2D given MW suggested lower risk for developing hypertension. Improved hepatic conditions were noted by decreases in alanine aminotransferase and aspartate aminotransferase among subjects with T2D given MW, indicating better insulin sensitivity and decreased tendency toward impaired liver function. CONCLUSION: Daily intake of 150 mL of MW or Dz-W with meals improved several metabolic responses among diabetics compared with diabetics given MJ.

Antioxidants↗
PubMed2006

Expression of the chalcone synthase gene from grape and preparation of an anti-CHS antibody.

Flavonoids are closely related to a plant's antioxidative ability. Because chalcone synthase (CHS) is the first enzyme to act as part of the flavonoid biosynthesis pathway, its expression and regulation are important. Here we present the expression of a full-length chs cDNA with 1225bp from grape seedlings as well as the preparation of an antibody against the expressed protein. A full-length chs cDNA was introduced into an expressed plasmid pET-30a(+) vector at the EcoRI and SalI restriction sites. pET-chs was found to be highly expressed in Escherichia coli BL21(DE3) pLysS cells with isopropyl-beta-d-thiogalactoside (IPTG) induction. A fusion protein with the His.tag label was purified by Ni-NTA His. Bind Resin and then used as the antigen to immunize a New Zealand rabbit. The resulting antiserum was then further precipitated by 50% saturated ammonium sulfate and DEAE-Sepharose FF column chromatography to obtain the immunoglobulin G (IgG) fraction. The resulting antibody was found capable of immuno-recognizing the CHS of the crude protein extracts from different grape tissues with a molecular mass of 43kDa.

Acyltransferases↗
PubMed2005

Protective effect of red grape seeds proanthocyanidins against induction of diabetes by alloxan in rats.

It has been documented that impaired homeostasis in diabetes mellitus is associated with increased production of reactive oxygen species and depletion of the antioxidant defense systems. Natural grape seed proanthocyanidins (GSP) are potent free radical scavengers and hence provide significant protection against oxidative stress. Accordingly, the present study focused on investigating the possible protective role of GSP against free radical-mediated damage in pancreatic tissues of alloxan-induced diabetes in rats. The results revealed that oral administration of 50 and 100 mg kg(-1) (body weight) of GSP for 72 h significantly increased pancreatic glutathione (GSH) levels and inhibited the increase in lipid peroxidation caused by alloxan (p < 0.001). On the other hand, a significant reduction in pancreatic total nitrate/nitrite content (p < 0.001) was observed. Furthermore, GSP caused significant decline in the hyperglycemia induced by alloxan (p < 0.001). Such antihyperglycemic effect of GSP was accompanied by a significant increase in serum insulin levels in diabetic rats following 72 h of administration (p < 0.001). In conclusion, the study suggests that GSP are effective in ameliorating the damage to pancreatic tissue in experimental diabetes mellitus. Such effect may be related to their potent antioxidant properties as evidenced by the increase in pancreatic GSH and reduction of lipid peroxidation as well as total nitrate/nitrite levels.

Alloxan↗
PubMed2006

Synergism and redundancy in a plant volatile blend attracting grapevine moth females.

A flight tunnel study was done to decipher the behavioral effect of grape odor in grapevine moth Lobesia botrana. A blend of 10 volatile compounds, which all elicit a strong antennal response, attracts mated grapevine moth females from a distance, by upwind orientation flight. These 10 grape volatiles are in part behaviorally redundant, since attraction to a 3-component blend of beta-caryophyllene, (E)-beta-farnesene and (E)-4,8-dimethyl-1,3,7-nonatriene was not significantly different from the 10-component blend. Blending these three compounds had a strong synergistic effect on female attraction, and omission of any one compound from this 3-component blend almost abolished attraction. It was nonetheless possible to substitute the three compounds with the other grape volatiles which are perceived by the female antenna, to partly restore attraction. Several blends, of varying composition, elicited significant attraction. The observed behavioral plasticity in response to grape volatile blends probably reflects the variation of the natural plant signal, since females oviposit on different grape varieties, in different phenological stages.

Animals↗
PubMed2006

The diversity of pathogenesis-related proteins decreases during grape maturation.

It was recently shown that wines contain typically a huge diversity of structurally similar polypeptides that exhibit a high degree of homology to pathogenesis-related (PR) proteins. This observation suggested the existence of one or a few precursors in mature grapes, common to most or all the wine PR proteins. Limited proteolysis and chemical modification of the precursor(s) during fruit ripening and winemaking could then generate the large number of distinct wine polypeptides. However, the patterns of PR proteins extracted from grape berries regularly harvested from the onset of development until maturity did not confirm the previous hypothesis. Two different methodologies, involving 2-D immunoblotting and a combination of FPLC cation/anion exchange chromatographies with 1-D immunoblotting, indicate that the total concentration of PR proteins is increased but its diversity is reduced from the early stages of berry development until maturity. These results indicate that PR proteins are synthesized in a wide variety of forms from the early stages of grape development, eliminating the hypothesis previously formulated on the existence of one or few precursors common to the wine proteins.

Amino Acid Sequence↗
PubMed2005

Antifungal effects of cysteine towards Eutypa lata, a pathogen of vineyards.

Cysteine inhibited mycelial growth of the pathogenic fungus affecting grapevines Eutypa lata Pers. Fr. Tul. and C. Tul. in a concentration-dependent manner. The threshold value (defined by the concentration inducing a growth inhibition higher than 5%) was 0.5 mM. A 10 mM concentration induced a complete inhibition of growth and triggered necrotic processes as evidenced by an increasing number of nuclei stained by propidium iodide. In conditions mimicking the plant environment (in particular, a pH near the apoplastic value, i.e. 5.5), 6 mM cysteine induced dramatic modifications in the structural organization of the mycelium (wall, mitochondria, vacuoles and nucleus) leading to death of the hyphae. The antifungal effect of the molecule increased at the acidic experimental pH (pH 4.1). The effect was highly specific to cysteine since modifying the molecular arrangement or masking the SH-function hindered the antifungal efficiency. Cysteine spectrum of action was broad among the various strains of E. lata tested. However, a lower efficiency was observed against fungal species intervening in other grapevine diseases (esca, black dead arm). Besides its direct antifungal effect, the role of cysteine presents particular interest in the fight against fungal pathogens since it triggered an excretion of ergosterol, a compound with elicitor properties. Therefore, cysteine may indirectly increase plant defense reactions.

Antifungal Agents↗
PubMed2005

DNA-based characterization of ochratoxin-A-producing and non-producing Aspergillus carbonarius strains from grapes.

Using molecular methods, a total of 189 strains of black aspergilli, including Aspergillus carbonarius and uniseriate species (Aspergillus aculeatus, Aspergillus japonicus), were studied in order to characterize species responsible for ochratoxin A (OTA) contamination of grapes from Europe and Israel. Sixty-six strains were morphologically identified as belonging to the uniseriate species and 123 as A. carbonarius. None of the uniseriate species were able to produce OTA. From the A. carbonarius strains, 96.7% were OTA producers (0.1-654.3 microg/g). We characterized 53 strains of A. carbonarius from different countries by RAPD and ITS-5.8S rDNA sequencing analysis. Forty-nine strains had a similar RAPD pattern and identical ITS-5.8S rDNA sequences. They produced OTA at different levels. No correlation was observed between the obtained clusters and the OTA production level or origin. Only four strains, morphologically identified as A. carbonarius, were unable to produce OTA. These strains showed a different RAPD pattern, and the section of DNA sequenced differed from the sequence of the other 49 strains. These OTA-non-producing strains may represent a new species in the Aspergillus section Nigri.

Aspergillus↗
PubMed2005

Mycotoxin-producing and other fungi isolated from grapes for wine production, with particular emphasis on ochratoxin A.

Mycotoxins are toxic secondary metabolites produced by filamentous fungi that have been detected in food commodities, including grapes and wine. A survey was conducted to assess mycotoxin-producing fungi in grapes destined for wine production. The mycotoxigenic capacity of the isolates was tested in culture media. Grapes were analyzed by plating methods from 4 Portuguese wine-growing regions at 3 maturation stages (pea berry, early veraison and ripe berry) between 2001 and 2003. From the 10,602 strains detected and identified, the most frequent genera were Cladosporium (25%), Alternaria (24%), Botrytis (15%), Penicillium (9%) and Aspergillus (8%). Most (92.0%) were non-mycotoxigenic or produced mycotoxins of unknown relevance to health. Potential producers of ochratoxin A (OTA) (Aspergillus niger aggregate, 5.4%, Aspergillus carbonarius, 0.6%) and trichothecenes (Fusarium spp., 0.4%; Trichothecium roseum, 0.8%) were the most frequent mycotoxigenic species isolated from grapes. OTA was detected in all cultures of A. carbonarius and 4% of A. niger aggregate strains. There was potential for OTA and trichothecene production in grapes by A. carbonarius and T. roseum, respectively, prior to harvest time. Data presented herein indicate that A. carbonarius is most likely to occur in vineyards with Mediterranean climates, while T. roseum is more likely to occur in more temperate climates, and is associated with gray rot. The present work emphasizes the need to use grapes under good condition so as to reduce the risk of contamination with mycotoxigenic fungi and subsequent mycotoxin occurrence in wine.

Aspergillus↗
PubMed2005

Assessment of copper bioavailability and toxicity in vineyard runoff waters by DPASV and algal bioassay.

The aim of this study was to assess the toxicity of runoff waters in an agricultural multipollution context through an in-depth assessment of copper bioavailability and toxicity. Runoff waters were screened for major ions, metals and diuron. The potential environmental impact of these runoff waters was evaluated using the conventional 72-h growth inhibition test with the green alga Pseudokirchneriella subcapitata. The results suggested that the toxicity detected in the calcareous vineyard field was due to the presence of diuron, whereas the non-calcareous runoff waters were non-toxic. Chemical speciation modelling by MINEQL revealed that most of the copper present in the non-toxic natural runoff waters was complexed by organic matter. These samples were spiked with copper, and then the toxicity and the electrochemically bioavailable copper fraction were measured. Differential pulse anodic stripping voltammetry (DPASV) was used to detect labile complexes and free copper. This combined approach highlighted the presence of some labile copper complexes in samples reaching the EC10-these could have contributed to the copper toxicity.

Agriculture↗
PubMed2005

Bioaerosol formation during grape stemming and crushing.

Indoor formation of airborne particles during pre-fermentation grape processing was assessed by particle counting using laser particle sizers. Particle numbers of four different aerodynamic size classes (0.3 to 0.5 microm, 0.5 to 1 microm, 1 to 5 microm, and >5 microm) were determined during unloading of harvest containers and subsequent grape stemming and crushing. Regarding these size classes, composition before grape handling was determined as 87.9%, 10.4%, 1.7%, and 0.1%, respectively, whereas the composition changed during grape handling to 50.4%, 15.2%, 33.0%, and 1.5%, respectively. Airborne bacteria and fungi originating from grape processing were collected by impactor and liquid impinger samplers. Grape handling resulted in a sixfold increase in total (biological and non-biological) airborne particles. The generation of bacterial and fungal aerosols was associated mostly with particles of aerodynamic diameters>5 microm (mainly 7 to 11 microm) as determined by flow cytometry. This fraction was increased 150fold in relation to background levels before grape crushing. Maximum concentrations of culturable bacteria reached 485,000 colony forming units (cfu/m3), whereas 146,000 cfu of fungi and yeasts were detected per cubic meter of air. Culturable Gram-negative bacteria occurred only in small numbers (180 cfu/m3). In relation to the total number of airborne particles emitted, culturable microorganisms comprised 0.1% to 0.2%. As soon as grape crushing was stopped, particle concentrations decreased rapidly either due to passive settling or due to air currents in the occupational indoor environment reaching background levels.

Aerosols↗
PubMed2005

Leaching of copper, chromium and arsenic from treated vineyard posts in Marlborough, New Zealand.

There have been conflicting reports as to the extent that copper-chromium-arsenic (CCA) treatments leach from timber. In New Zealand, vineyards utilise CCA-treated posts at a rate of 579 posts per hectare. This represents a potential CCA burden on the soil of 12, 21, and 17 kg/ha, respectively, for the three elements. Given a replacement rate of 4% per year, the use of CCA-treated posts may result in an accumulation of these elements in the soil, possibly leading to groundwater contamination. We undertook a general survey to determine the extent of CCA leaching from treated vineyard posts. Treated Pinus radiata posts were sampled at six sites around the Marlborough region of New Zealand to represent a range of post ages and soil types. For each post, above- and belowground wood samples were taken. As well, the soil adjacent to the post was sampled at a 50 mm horizontal and 100 mm vertical distances from the post. The belowground wood samples of the posts had significantly lower CCA concentrations than the aboveground portions, which were not significantly different from new posts. This indicates leaching. Soils surrounding the posts had significantly higher CCA concentrations than control soils. Higher CCA concentrations were measured under the posts than laterally. Some 25% of the samples exceeded 100 mg/kg As, the Australian National Environment Protection Council (ANEPC) guideline level for As in agricultural soil, and 10% exceeded 100 mg/kg Cr, the ANEPC limit for chromium. At one site, we found a significant positive correlation between post age and CCA-leaching. The CCA issue could be eliminated by using alternative posts, such as steel, concrete, or untreated woods such as Eucalyptus or beech. Alternatively, CCA-treated posts could, for example, be lacquered or otherwise protected, to reduce the rate of CCA leaching.

Agriculture↗
PubMed2005

Anti-thrombotic effect of proanthocyanidin, a purified ingredient of grape seed.

INTRODUCTION: Moderate and regular consumption of wine reduces the risk of acute coronary thrombotic events. The mechanism of the anti-thrombotic effect of wine is not clear. Extract or purified ingredients of grapes have not yet been studied for anti-thrombotic effect. MATERIALS AND METHODS: Anti-thrombotic effect of proanthocyanidin, a highly purified ingredient of grape seed, was assessed by a shear-induced thrombosis test in vitro and by a laser-induced thrombosis test in the mouse carotid artery, in vivo. RESULTS AND CONCLUSIONS: Intravenously (20 mg/kg body weight, BW) or orally (2 x 200 mg/kg BW) administered proanthocyanidin significantly inhibited the laser-irradiation induced thrombus formation in the carotid artery (both P=0.01). Subsequent to oral administration of proanthocyanidin, in vitro platelet reactivity to shear stress has been inhibited. The latter suggests that the in vivo anti-thrombotic effect of proanthocyanidin may be due to a direct inhibitory effect on platelets.

Animals↗
PubMed2004

Engineering grapevine for increased resistance to fungal pathogens without compromising wine stability.

The vast majority of wine proteins have recently been identified as pathogenesis-related (PR) proteins. During the growing season, these proteins are expressed in developmentally dependent and inducible manners in grapevine leaves and grape berries, in which they are believed to play an important role in protection against fungal pathogens and possibly other stresses. Because of their inherent resistance to proteolytic attack and to the low pH values characteristic of wines, vinification can be seen as a "purification strategy" for grape PR proteins. The inevitable consequent accumulation of these proteins in wines becomes a technological nuisance because they adversely affect the clarity and stability of wines. Genetically modified vines underexpressing PR proteins would certainly lead to stable wines but would increase the plant susceptibility to fungal attack, and the actual trend seems to be in the opposite direction, that is overexpressing these proteins to obtain plants with enhanced resistance to pathogens--a trend that will probably augment problems associated with protein instability in the resulting wines.

Fungi↗
PubMed2006

In vitro protective effect of a Jacquez grapes wine extract on UVB-induced skin damage.

Several studies have shown that UV radiation on the skin results in the formation of reactive oxygen species (ROS) that interact with proteins, lipids and DNA, thus altering cellular functions. The epidermis is composed mainly of keratinocytes, rich in ROS detoxifying enzymes and in low-molecular-mass antioxidant molecules. However, the increased generation of ROS can overwhelm the natural defences against oxidative stress. Therefore treatment of the skin with products containing plant-derived antioxidant ingredients may be a useful strategy for the prevention of UV-mediated cutaneous damage. In the present study we have investigated the in vitro capability of a Jacquez grapes wine extract (containing a significant level of proanthocyanidins, together with lower amounts of anthocyanins and hydroxycinnamic acids; JW-E), to protect skin against UVB-induced oxidative damage by using a three-dimensional tissue culture model of human epidermis. The endpoints of our experiments were cell viability, release of interleukin-1alpha and prostaglandin E(2) (well-known mediators of cutaneous inflammatory processes), accumulation in the epidermis of malondialdehyde/4-hydroxynonenal and protein carbonyl groups (derived by the oxidative damage respectively of lipids and proteins) and tissue redox balance (expressed by the levels of reduced glutathione, oxidized glutathione, glutathione peroxidase and glutathione reductase). Taken together, our findings demonstrate that the JW-E is an efficient botanical mixture able to prevent skin oxidative damage induced by UV-B exposure and may thus be a potential promising candidate as a skin photoprotective agent.

Aldehydes↗
PubMed2006

Comparative efficacy of oligonol, catechin and (-)-epigallocatechin 3-O-gallate in modulating the potassium bromate-induced renal toxicity in rats.

Potassium bromate (KBrO(3)) is a by-product from ozonation of high-bromide surface water for production of drinking water and is a rodent carcinogen. Oligonol is a product emanating from the oligomerization of polyphenols, typically proanthocyanidin from a variety of fruits (grapes, apples, persimmons, etc.) and contains catechin-type monomers and proanthocyanidin oligomers. In this study, the ability of oligonol derived from grape seeds, grape seeds extracts (Product A, containing biologically active flavonoids and the oligomeric proanthocyanidin) and pine bark extracts (Product B, composed of flavan-3-ol derivatives) to modulate the KBrO(3)-induced renal toxicity was compared with (+) catechin and (-)-epigallocatechin 3-O-gallate (EGCG). In the Trolox equivalent antioxidant capacity (TEAC) assay, the order of the antioxidant activity was EGCG>catechin>oligonol>Product A>Product B. However, oligonol elicits the strongest antioxidant capacity following in vivo supplementation to rats, with the order of efficacy of oligonol>Product A> or =Product B>EGCG>catechin. Blood levels of lipid peroxidation products (LPO), urea nitrogen (BUN) and creatinine were elevated by KBrO(3) treatment. Oligonol significantly restored LPO to the level in the untreated rats and had the strongest potency when compared with the effects of Products A and B. The five materials lowered KBrO(3)-induced BUN level, but this was not statistically significant. Oligonol significantly reduced the increased level of the creatinine, seconded by Product A, Product B and EGCG. Catechin had the lowest effect in both the BUN and creatinine levels. That oligonol was able to modulate KBrO(3)-induced lipid peroxidation and the levels of blood urea nitrogen and creatinine suggests potential chemopreventive function and application in mitigating toxicity due to long-term exposure to KBrO(3) in public drinking water.

Animals↗
PubMed2005

Effect of ethanol on protection of urinary bladder function by grape suspensions.

OBJECTIVES: To compare the protective effects of grape suspensions prepared in an aqueous vehicle with grape suspensions prepared in an 8% ethanol vehicle in rabbits subjected to partial outlet obstruction. The hypothesis was that the presence of ethanol would increase the absorption of the beneficial components of the grape suspensions and thus increase their protective ability. The use of ethanol in these studies was not to simulate wine. METHODS: A total of 48 New Zealand white rabbits were separated into eight groups of 6 rabbits each. Groups 1 and 3 were pretreated by oral gavage for 3 weeks with grape suspensions in water; groups 2 and 4 were treated with vehicle. Groups 5 and 7 were treated with the grape suspensions in 8% ethanol, and groups 6 and 8 were treated with ethanol vehicle. Groups 1, 2, 5, and 6 underwent sham operations, and groups 3, 4, 7, and 8 underwent partial outlet obstruction. Three weeks after surgery, the rabbits were evaluated. RESULTS: The bladder weight had significantly increased in all obstructed groups. The contractile responses to field stimulation and carbachol were reduced in all obstructed groups, although the responses in both grape-treated groups were greater than both vehicle-treated groups. The contractile responses to potassium chloride were significantly reduced by partial outlet obstruction in both obstructed groups similarly. CONCLUSIONS: Both grape suspensions provided protection against obstructive-induced bladder dysfunction. The ethanol preparation of the grape suspension was not better than the aqueous preparation.

Animals↗
PubMed2005

Occurrence of a DNA sequence of a non-retro RNA virus in a host plant genome and its expression: evidence for recombination between viral and host RNAs.

This study demonstrates that sequences homologous to those of the non-retro RNA virus (Potato virus Y; PVY) are integrated into the genome of several grapevine varieties. The integrated PVY-coat-protein-like cistron is expressed in the grapevine as indicated by Southern and Western blot analyses as well as by RNase protection assay. In addition, genome-walking studies showed that one PVY-like sequence is flanked by 41-bp direct repeats and is embedded in authentic grapevine sequences, flanked by inverted repeats. Rearranged PVY-like sequences were also found in tobacco. It is suggested that nonhomologous recombination of a potyviral RNA with RNA of a retrotransposable element took place at some point in evolution. The initial integration locus was probably within a grapevine gene homologous to a pentatricopeptide repeat-carrying protein, and was later transposed to other locations. The current location is reminiscent of a MITE-type retroelement, indicating transposition history. Because grapevine cultivars are propagated asexually, without going through a meiotic phase, the chance for DNA recombination is minimal and the foreign integrated sequence may be better conserved, enabling it to be expressed correctly in the recipient genome.

Amino Acid Sequence↗