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Influence of micro-oxygenation treatment before oak aging on phenolic compounds composition, astringency, and color of red wine.

Micro-oxygenation is usually applied to red wines as a cheaper alternative to oak aging. It has been suggested, however, that micro-oxygenation can also be used to complement oak aging in order to improve the quality of very astringent and herbaceous red wines. In this paper we study how applying the micro-oxygenation technique before oak aging affects the composition and quality of astringent red wines. When this technique is applied prior to oak aging, the wines have a slightly less intense red color and significantly higher levels of combined and free anthocyanins and ethyl-bridged anthocyanin-flavanol pigments. On the other hand, no differences in other newly formed pigments are found. Applying micro-oxygenation before oak aging does not affect the total proanthocyanidin concentration, but it produces wines with a slightly (though significantly) higher mean degree of proanthocyanidin polymerization and a drastically lower astringency. These wines also present a clearer impact of wood aromas.

Anthocyanins↗

Sorption behavior of volatile phenols at the oak wood/wine interface in a model system.

The sorption in a model system of aroma compounds of enological interest (mixture of the eight derivatives from guaiacol, 4-ethylphenol, and whiskylactone) onto wood was investigated to assess the influence of wood on the concentration of these volatiles during the aging of wine. To evaluate the influence of the solubility of aroma compounds in sorption phenomena, this parameter was determined for each volatile compound in model wine at 10 and 25 degrees C. The solubility is significantly higher in the model wine than in water and remains constant in the range of temperatures studied, except for guaiacol and vanillin. Kinetic and equilibrium sorptions were investigated. Sorption kinetics showed that the sorption equilibrium for all aroma compounds was reached after 6-7 days. From the study of the individual sorption isotherms, two distinct kinds of sorption behavior were observed depending on the presence or not of an ethylenic para substituent conjugated to the phenyl ring. K(ww) partition coefficients between the wood and the model wine were determined, which exhibited an unusual positive variation with temperature.

Adsorption↗

Volatile composition and contribution to the aroma of spanish honeydew honeys. Identification of a new chemical marker.

Honeydew honeys from holm-oak, oak, and forest were isolated for aroma compounds by simultaneous distillation-extraction and analyzed by gas chromatography-mass spectrometry. In all, 66 volatile components were identified and quantified. trans-Oak lactone, a characteristic volatile component of oak wood, is proposed as a new chemical marker for the plant origin of honeydew honeys. Other compounds, such as aminoacetophenone and propylanisol, could be considered characteristic of holm-oak honeydew honeys. A total of 15 volatile compounds presented odor activity values (OAVs) greater than 1, with phenylacetaldehyde and beta-damascenone being those with the highest OAVs.

Acetaldehyde↗

Aroma extraction dilution analysis of Sauternes wines. Key role of polyfunctional thiols.

The aim of the present work was to investigate Sauternes wine aromas. In all wine extracts, polyfunctional thiols were revealed to have a huge impact. A very strong bacon-petroleum odor emerged at RI = 845 from a CP-Sil5-CB column. Two thiols proved to participate in this perception: 3-methyl-3-sulfanylbutanal and 2-methylfuran-3-thiol. A strong synergetic effect was evidenced between the two compounds. The former, never mentioned before in wines, and not found in the musts of this study, is most probably synthesized during fermentation. 3-Methylbut-2-ene-1-thiol, 3-sulfanylpropyl acetate, 3-sulfanylhexan-1-ol, and 3-sulfanylheptanal also contribute to the global aromas of Sauternes wines. Among other key odorants, the presence of a varietal aroma (alpha-terpineol), sotolon, fermentation alcohols (3-methylbutan-1-ol and 2-phenylethanol) and esters (ethyl butyrate, ethyl hexanoate, and ethyl isovalerate), carbonyls (trans-non-2-enal and beta-damascenone), and wood flavors (guaiacol, vanillin, eugenol, beta-methyl-gamma-octalactone, and Furaneol) is worth stressing.

Chromatography, Gas↗

Triterpenic and other lipophilic components from industrial cork byproducts.

The detailed chemical composition of the lipophilic extractives of cork and cork byproducts generated throughout industrial processing has been investigated by gas chromatography-mass spectrometry. Triterpenes (cerine, friedeline, and betulinic acid) were the major components detected. Betulinic acid is the main triterpene (11.7 g/kg) identified in industrial cork powder, whereas in black condensates friedeline (95.3 g/kg), betuline (13.1 g/kg), and betulinic acid (12.1 g/kg) are the main triperpenes. Significant fractions of alpha-hydroxy fatty acids (115.1 g/kg) and alpha,omega-dicarboxylic acids (21.2 g/kg) were also detected in black condensate after alkaline hydrolysis. The results demonstrate that these two industrial byproducts can be considered as promising sources of bioactive chemicals or chemical intermediates for the synthesis of polymeric materials.

Gas Chromatography-Mass Spectrometry↗

Characterization of volatiles in extracts from oak chips obtained by accelerated solvent extraction (ASE).

Sixteen commercially available oak chips, differing in origin (French or American) and toasting level, were extracted by an accelerated solvent extraction method and characterized by their volatile composition. About 80 compounds were identified and quantified, a great part of them from the thermodegradation of lignin and cellulose. One furanone (solerone) and two C-13 norisoprenoids (3-oxo-alpha-ionol and a 3-oxoretro-alpha-ionol isomer) were also tentatively identified and reported for the first time in toasted wood. Quantitative data demonstrated the oak chips to be not so different from the composition of light- or medium-toasted wood barrels, which was reported by other authors. The same data suggest that toasting level had the strongest influence on the volatile composition of chip samples. Phenyl ketones, volatile phenols, and some furanic compounds were the most influenced. On the other hand, the influence of wood origin was found to be weaker.

Cellulose↗

Extraction, detection, and quantification of flavano-ellagitannins and ethylvescalagin in a Bordeaux red wine aged in oak barrels.

An extraction procedure and an analytical method have been developed to detect and quantify for the first time a series of ellagitannin derivatives formed in wine during aging in oak barrels. The method involves a preliminary purification step on XAD7 HP resin followed by a second purification step on TSK 40 HW gel. The resulting extract is analyzed for compound identification and quantitative determination by high-performance liquid chromatography-electrospray ionization-mass spectrometry in single ion recording mode. Reference compounds, which are accessible through hemisynthesis from the oak C-glycosidic ellagitannin vescalagin, were used to build calibration curves, and chlorogenic acid was selected as an internal standard. This method enabled us to estimate the content of four flavano-ellagitannins and that of another newly identified wine polyphenol, beta-1-O-ethylvescalagin, in a Bordeaux red wine aged for 18 months in oak barrels. All five ellagitannin derivatives are derived from the nucleophilic substitution reaction of vescalagin with the grape flavan-3-ols catechin and epicatechin or ethanol.

Biphenyl Compounds↗

Synthetic secofriedelane and friedelane derivatives as inhibitors of human lymphocyte proliferation and growth of human cancer cell lines in vitro.

Controlled silylation of friedelin (1) from cork smoker wash solids, a byproduct generated during processing of corkboard by steam baking, gave 3-trimethylsiloxyfriedel-2-ene (3) in high yields. Oxidation of 3 with OsO(4)/NMMO produced 2alpha-hydroxyfriedelan-3-one (cerin) (5), from which the new 2,3-secofriedelan-2-al-3-oic acid (6) was obtained quantitatively by periodic acid oxidation. Oxidation of 3 with DDQ afforded friedel-1-en-3-one (8). Reductive ozonolysis of 3 gave 2alpha,3beta-dihydroxyfriedelane, pachysandiol A (7). Compound 6 proved to be a potent inhibitor of human lymphocyte proliferation (IC(50) = 10.7 microM) and of the growth of a human cancer cell line (GI(50) = 5.4-17.2 microM). (13)C NMR data for compounds (3, 4, 5, 6a,7, and 8) are described for the first time.

Antineoplastic Agents, Phytogenic↗

Biscopyran, a phytotoxic hexasubstituted pyranopyran produced by Biscogniauxia mediterranea, a fungus pathogen of cork oak.

A new phytotoxic hexasubstituted pyranopyran, biscopyran (3), was isolated together with phenylacetic acid (2) and previously isolated 5-methylmellein (1) from the liquid culture filtrates of Biscogniauxiamediterranea, a major fungal pathogen involved in oak decline in Sardinia. Biscopyran was characterized by spectroscopic methods as a new (Z)-2-methoxy-1-[7-((Z)-2-methoxybut-2-enoyl)-3,4,5,6-tetramethyl-2H,7H-pyrano[2,3-b]pyran-2-yl]but-2-en-1-one. Biscopyran assayed at 0.26-0.026 mM concentration range caused epinasty on cork oak cuttings. On a nonhost plant, tomato, biscopyran caused wilting. Phenylacetic acid, assayed at the same concentration, was toxic to Q. suber, while on tomato cuttings it induced internal tissue collapse on the stem.

Fungi↗

Utilization of a 1,2-dioxine for the synthesis of the four possible stereoisomers of oak lactone.

[reaction: see text]. The natural products cis- and trans-oak lactone (1) have been prepared, along with their enantiomeric counterparts, from furanone 12, which was itself prepared from racemic 1,2-dioxine 9 and a chiral malonate diester. The key steps in the synthesis of 1 are the use of the malonate diester as a chromatographic resolving agent and the decarboxylation of 13, which can be directed to give either the cis- or trans-product. This leads to all four possible oak lactone stereoisomers from a common intermediate.

Biological Products↗

Contaminating resin acids have not caused the high rate of sensitivity to oak moss.

Commercially available oak moss absolute patch test material has recently been shown to contain resin acids of the type found in colophony (colophonium). We wished to assess whether the high frequency of positive patch tests to oak moss absolute at this institute was likely to reflect significant contamination by resin acids. The rate of positive reactions to colophony among our oak moss-allergic patients patch tested during 1984-2000 was retrospectively investigated. 25395 patients were tested to the European standard series during this period and 1963 (7.7%) were allergic to the fragrance mix. 342 of these patients were allergic to oak moss absolute, of whom 73 (21.3%) were allergic to colophony. In comparison, 115 (13.4%) of 861 fragrance mix-positive but oak moss-negative patients were allergic to colophony (p = 0.0002, Fisher's exact test). This strongly statistically significant association between oak moss absolute and colophony shows only a small increase in rates of allergy to colophony in oak moss-positive patients. Thus we conclude that contaminating resin acids have not alone caused the high rate of sensitivity to oak moss.

Acids↗

An investigation into effects of long-distance seed dispersal on organelle population genetic structure and colonization rate: a model analysis.

A simulation-based modelling approach is used to examine the effects of stratified seed dispersal (representing the distribution of the majority of dispersal around the maternal parent and also rare long-distance dispersal) on the genetic structure of maternally inherited genomes and the colonization rate of expanding plant populations. The model is parameterized to approximate postglacial oak colonization in the UK, but is relevant to plant populations that exhibit stratified seed dispersal. The modelling approach considers the colonization of individual plants over a large area (three 500 km x 10 km rolled transects are used to approximate a 500 km x 300 km area). Our approach shows how the interaction of plant population dynamics with stratified dispersal can result in a spatially patchy haplotype structure. We show that while both colonization speeds and the resulting genetic structure are influenced by the characteristics of the dispersal kernel, they are robust to changes in the periodicity of long-distance events, provided the average number of long-distance dispersal events remains constant. We also consider the effects of additional physical and environmental mechanisms on plant colonization. Results show significant changes in genetic structure when the initial colonization of different haplotypes is staggered over time and when a barrier to colonization is introduced. Environmental influences on survivorship and fecundity affect both the genetic structure and the speed of colonization. The importance of these mechanisms in relation to the postglacial spread and genetic structure of oak in the UK is discussed.

Climate↗

Determination of gallic acid in wood dust as an indicator of oak content.

A high-performance liquid chromatography (HPLC) method was developed for the detection of extracted gallic acid in wood dust. Gallic acid is a polyphenol present in carcinogenic oak wood dust, but not in beech, ash, pine or spruce dusts, as confirmed by HPLC analyses. The method involved the extraction of gallic acid from the oak dust, followed by liquid chromatographic analysis. The correlation coefficient for the share of oak dust vs. the gallic acid concentration of wood dust was 0.995. The method was tested with oak wood dust samples collected on polycarbonate membrane filters during an 8 h workshift in a floor board factory, where the dust content of the air samples was determined gravimetrically. The oak dust and the gallic acid concentrations varied from 0.2 to 13.8 mg m-3 and from 0.03 to 3.8 micrograms m-3, respectively. These parameters were linearly correlated with a correlation coefficient of 0.95. The airborne gallic acid determination is a useful technique to confirm occupational exposure to oak wood dust, a recognized human carcinogen.

Carcinogens↗

Fagales pollen sensitization in a birch-free area: a respiratory cohort survey using Fagales pollen extracts and birch recombinant allergens (rBet v 1, rBet v 2, rBet v 4).

BACKGROUND: Birch allergy is one of the most common pollinosis in areas where exposure to high levels of birch pollen is common. Little is known about birch sensitivity in areas without birch pollen exposure and reactivity to birch-related species within the Fagales order. OBJECTIVE: the aim was to evaluate Fagales reactivity within a population not exposed to birch pollen using epidemiological, diagnostic, and laboratory approaches by means of extracts and allergenic molecules. METHODS: A cohort of 5335 respiratory allergic patients was screened by means of skin testing birch, hazel, and oak pollen extracts. Patients were from a birch-free area, but exposed to other Fagales pollen species. A subset of patients was from an intensively cultivated hazel area. A sample of the Fagales allergic population was tested with other Fagales pollen extract (alder, hornbeam, beech, chestnut) and with apple and hazelnut. IgE detection was performed with birch, hazel, oak, apple, and hazelnut extracts, and with Bet v 1, Bet v 2, Bet v 4, and bromelain. IgE immunoblots were performed using birch and hazel extracts. Epidemiological, clinical, and laboratory data were analysed by stratifying the allergic population. RESULTS: Twenty-five percent of the pollen allergic cohort was skin test positive to at least one of the three Fagales species. Combined reactivity to the three species was recorded in 80% of this cohort. Isolated hazel pollen reactivity was recorded in 13.5% of the Fagales allergic patients. Sixty-six percent of these subjects were from the intensively cultivated hazel area. Reactivity to apple and hazelnut was detected by skin test (40%) and IgE reactivity (60%), but only 19% of the positive patients reported symptoms related to at least one of the two foods. Reactivity to Bet v 1 was recorded in 84% of the birch/hazel/oak co-reactivity group, and in 28% of the subjects with the same co-reactivity, but associating a multiple pollen sensitization. IgE to Bet v 2 (50%) and Bet v 4 (23%) panallergens were recorded positive in the latter subset. Bet v 1 prevalence ranged between 48% and 21% among subgroups of patients coming from different areas. Furthermore, an IgE reactivity to hazel-restricted allergenic components was detected among subjects coming from the same area and having a hazel isolated reactivity. CONCLUSION: Fagales allergy can be found in birch-free areas caused by the exposure to other Fagales species. Birch allergens can be useful for mimicking the allergenic extract, but are also the exclusive tools for a fine diagnostic and epidemiological approach to Fagales pollen allergy. Allergenic molecules from the hazel family will increase the panel of available reagents for the molecule-based approach to allergy diagnosis and therapy.

Adult↗

The incidence and diversity of Wolbachia in gallwasps (Hymenoptera; Cynipidae) on oak.

Wolbachia bacteria infect approximately 20% of all insect species, and cause a range of alterations to host reproduction, including imposition of thelytoky. The incidence and phenotypic impact of Wolbachia remains to be established in many insect taxa, and considerable research effort is currently focused on its association with particular reproductive modes and the relative importance of the various pathways via which infection occurs. Gallwasps represent an attractive system for addressing these issues for two reasons. First, they show a diversity of reproductive modes (including arrhenotoky, thelytoky and cyclical parthenogenesis) in which the impact of Wolbachia infection can be examined. Second, they occupy two intimately linked trophic niches (gall-inducers and inquilines) between which there is potential for the horizontal exchange of Wolbachia infection. In the arrhenotokous gallwasp lineages screened to date (the herb-galling 'Aylacini' and the rose-galling Diplolepidini), Wolbachia infection always induces thelytoky. The impact of Wolbachia in other arrhenotokous clades, and in the cyclically parthenogenetic clades remains unknown. Here we use polymerase chain reaction (PCR) screening and sequence data for two Wolbachia genes (wsp and ftsZ) to examine the prevalence and incidence of Wolbachia infection in 64 species (a total of 609 individuals) in two further tribes: the arrhenotokous inquilines (tribe Synergini), and the cyclically parthenogenetic oak gallwasps (tribe Cynipini). We ask: (i) whether Wolbachia infection has any apparent impact on host reproduction in the two tribes and (ii) whether there is any correlation between Wolbachia infection and the apparent lack of an arrhenotokous generation in many oak gallwasp life cycles. We show: (i) that Wolbachia infection is rare in the Cynipini. Infected species show no deviation from cyclical parthenogenesis, and infection is no more common in species known only from a thelytokous generation; (ii) that there is a higher incidence of infection within the arrhenotokous inquilines, and generally in gallwasp tribes without cyclical parthenogensis; (iii) all Wolbachia-positive inquiline species are known to possess males, implying either that Wolbachia infection does not result in loss of sex in this tribe or, more probably, that (as for some rose gallwasps) Wolbachia infection leads to loss of sex in specific populations; and (iv) although we find some inquilines and gall inducers to be infected with Wolbachia having the same wsp sequence, these hosts are not members of the same gall communities, arguing against frequent horizontal transmission between these two trophic groups. We suggest that exchange may be mediated by the generalist parasitoids common in oak galls.

Animals↗

Checking the geographical origin of oak wood: molecular and statistical tools.

New methods for better identification of timber geographical origin would constitute an important technical element in the forest industry, for phytosanitary certification procedures or in the chain of custody developed for the certification of timber from sustainably managed forests. In the case of the European white oaks, a detailed reference map of chloroplast (cp) DNA variation across the range exists, and we propose here to use the strong geographical structure, characterized by a differentiation of western vs. eastern populations, for the purpose of oak wood traceability. We first developed cpDNA markers permitting the characterization of haplotype on degraded DNA obtained from wood samples. The techniques were subsequently validated by confirming the full correspondence between genotypes obtained from living tissues (buds) and from wood collected from the same individual oak. Finally, a statistical procedure was used to test if the haplotype composition of a lot of wood samples is consistent with its presumed geographical origin. Clearly, the technique cannot permit the unambiguous identification of wood products of unknown origin but can be used to check the conformity of genetic composition of wood samples with the region of alleged origin. This could lead to major applications not only in the forest industry but also in archaeology or in palaeobotany.

DNA Primers↗

Effect of dietary copper and vitamin E supplementation, and extensive feeding with acorn and grass on longissimus muscle composition and susceptibility to oxidation in Iberian pigs.

The objective of this study was to assess the effect of dietary copper and/or vitamin E supplementation on composition and oxidation of M. longissimus in Iberian pigs and to compare it with meat samples from pigs produced extensively and fed with acorn and grass. Grass had the highest alpha-tocopherol content (> 150 mg/kg DM), while acorns had the highest copper concentration (78.1 mg/kg DM). Dietary treatment did not affect copper composition in muscle and no interactions were detected between copper and alpha-tocopherol. The alpha-tocopherol content in muscle from pigs fed diets supplemented with alpha-tocopheryl acetate (100 mg/kg feed) was 1.5 times greater (p < 0.0001) than those from pigs receiving a basel diet. M. longisssimus dorsi from pigs fed extensively had a higher concentration of alpha-tocopherol than those fed in confinement with the basel level of alpha-tocopheryl acetate, but lower values than pigs fed supplemented levels. Total n-3 fatty acids (p < 0.02) and hematin (p < 0.0001) concentrations were significantly higher in muscle from pigs fed extensively than when fed in confinement. Muscle homogenates from Iberian pigs fed in extensive conditions showed significantly (p < 0.02) higher susceptibility to oxidation than those from pigs fed mixed diets. Dietary alpha-tocopheryl acetate supplementation (100 mg/kg feed) significantly (p < 0.05) reduced lipid oxidation of muscle, while dietary copper concentration did not modify susceptibility to lipid oxidation.

Animal Feed↗