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Biosynthesis of straight-chain ester volatiles in red delicious and granny smith apples using deuterium-labeled precursors.

Biosynthesis of straight-chain ester volatiles by Granny Smith and Red Delicious apples was investigated using deuterium-labeled fatty acids, C-6 aldehydes, and alcohols. Perdeuterated saturated and monounsaturated fatty acids were metabolized to hexyl-d(11), hexanoate-d(11), heptanoate-d(13), and octanoate-d(15) esters, whereas perdeuterated linoleic acid produced only hexyl-d(11) and hexanoate-d(11) esters. Exposure of fruit to vapors of deuterated 3Z-hexenal, 2E-hexenal, and hexanal identified the following biosynthetic processes: (1) isomerization between 3E, 3Z, and 2E-hexenals; (2) reduction to 3E, 3Z, and 2E-hexenyl esters; (3) reduction to hexanol and hexyl esters; (4) oxidation to hexanoic acid and formation of hexanoate esters; (5) beta-oxidation of hexanoic acid leading to butyl and butanoate esters; and (6) alpha-oxidation of hexanoic acid leading to pentyl and pentanoate esters. Unsaturated straight-chain ester volatiles appear to arise only by the lipoxygenase pathway and may be useful indicators of lipoxygenase activity in fruit.

Chromatography, Gas↗

Multivariate analysis of superficial scald susceptibility on Granny Smith apples dipped with different postharvest treatments.

Multivariate methods were used to present the results of different postharvest treatments applied to control superficial scald disorder in Granny Smith apples. In a two-season study, the treatments applied by dips were diphenylamine (DPA), Semperfresh (a sucrose ester coating) formulated alone or with either ascorbyl palmitate or n-propyl gallate, and CaCl(2). DPA correlated negatively with scald, whereas the rest of the treatments showed no clear relationship with the disorder. By means of partial least squares, the incidence of superficial scald after cold storage was shown to correlate with quality parameters (firmness and acidity), measured at the end of storage. In contrast, ethylene production did not appear as a relevant factor in the incidence of scald.

Cold Temperature↗

Impact of 1-methylcyclopropene and methyl jasmonate on apple volatile production.

Climacteric Fuji apples were treated with 10 microL x L(-1) MCP (1-methylcyclopropene), 2 mmol x L(-1) MJ (methyl jasmonate), or a combination of 10 microL x L(-1) MCP and 2 mmol x L(-1) MJ. Fruit were kept at 20 degrees C for 15 days after treatment. Production of ethylene and other volatile compounds was measured prior to and 3, 7, 11, and 15 days after treatment. Ethylene production decreased 3 days following MJ treatment and then increased. MCP treatment alone or in combination with MJ inhibited ethylene production. MJ and MCP inhibited production of many volatile alcohols and esters. The production of individual alcohols and esters appears to be differentially inhibited by MJ or MCP. MJ and MCP inhibited not only production of alcohols but also formation of esters from alcohols.

Acetates↗

Study of the phenolic profile of cider apple cultivars at maturity by multivariate techniques.

Phenolic compounds in 46 Spanish cider apple varieties were determined by RP-HPLC with direct injection. Several pattern recognition procedures, including principal component analysis (PCA), linear discriminant analysis (LDA), and partial least squares (PLS-1), were applied to the data in an attempt to classify the samples into bitter and nonbitter categories. Reliable decision rules were obtained by both LDA and PLS-1. LDA achieved 91.3 and 85.7% correct classification respectively, for internal and external evaluation of the model.

Beverages↗

Effect of hexanal on the shelf life of fresh apple slices.

In this work the effects of hexanal, as a component of packaging atmosphere, on the shelf life of and evolution of naturally occurring microbial populations in fresh apple slices during storage at 4 and 15 degrees C were evaluated. Although hexanal had no bactericidal effects, in all conditions considered, this volatile molecule significantly extended the shelf life. In fact, the presence of hexanal in the storage atmosphere (at 4 degrees C) totally inhibited mesophilic bacteria and considerably prolonged the lag phase of psychrotrophic bacteria. Also, at 15 degrees C, hexanal strongly inhibited molds, yeasts, and mesophilic and psychrotrophic bacteria. Moreover, hexanal led to a yeast selection favoring species having a reduced spoilage potential due to their prevalent respiratory activity. When added to a modified atmosphere (70% N(2) and 30% CO(2)), this molecule was also very effective in preventing browning reactions for at least 16 days at 15 degrees C. No changes in hue angle values were observed in samples packaged in modified atmosphere with hexanal, even after 16 days of storage at 4 degrees C.

Aldehydes↗

Migration of organophosphorus insecticides cyanophos and prothiofos residues from impregnated paper bags to Japanese apple-pears (Pyrus pyrifolia nakai cv. Nijisseiki).

Bags impregnated with the organophosphorus pesticides prothiofos and cyanophos in three levels were used to cover Japanese apple-pears to protect them from insects. The amounts of prothiofos residue in the bags collected 4 months after application ranged from 0 to 34% of the amounts (81, 148, and 333 microg) found in the bags prior to a bagging. The amounts of cyanophos residue in bags collected 4 months after application ranged from 11 to 29% of the amounts (335, 2860, and 3740 microg) present prior to a bagging. Amounts of prothiofos found in fruits after 4 months ranged from 0 to 0.076 ppm. Amounts of cyanophos found in fruits after 4 months ranged from 0 to 0.011 ppm. Results of the present study indicate that these two pesticides migrated from pesticide-impregnated bags to fruits during the growing season.

Food Contamination↗

Enantiomeric synthesis of (S)-2-methylbutanoic acid methyl ester, apple flavor, using lipases in organic solvent.

Enantiomeric selective synthesis of (S)-2-methylbutanoic acid methyl ester, which is known as a major apple and strawberry flavor, was performed from racemic 2-methylbutanoic acid using lipases in organic solvent. Among 20 lipases, lipase IM 20 (immobilized lipase of Rhizomucor miehei), lipase AP (Aspergillus niger), and lipase FAP-15 (Aspergillus javanicus) exhibited higher enzymatic activities and enantioselectivities and were selected for the synthesis of (S)-2-methylbutanoic acid methyl ester. Using these enzymes, the reaction conditions such as temperature and lyophilizing pH were optimized, and kinetic parameters were determined. All of the reactions were performed in isooctane, which was identified as the best reaction media for nonaqueous systems. At 20 degrees C maximum enantiomeric excess was observed, while synthetic activity increased as the temperature increased. Only lipases lyophilized at pH 5.5, 6. 0, 6.5, and 7.0 showed synthetic activity. In this pH range, enantioselectivities were not influenced by the lyophilizing pH. The K(M,S) and K(M,R) values for ester synthetic activity of lipase were 1120 and 1240 mM, respectively. Enzyme activity was inhibited by (S)-2-methylbutanoic amide, and its K(i) was calculated as 84 mM. (S)-2-Methylbutanoic amide acted as a competitive inhibitor.

Butyrates↗

Assimilation of volatiles from ripe apples by Sporidiobolus salmonicolor and Tilletiopsis washingtonensis.

Sporidiobolus salmonicolor ATCC 623 and Tilletiopsis washingtonensis NRRL Y-2555 grew on carbon resources provided as volatiles by ripe 'Golden Delicious' apples. This ability was not correlated with the reported natural habitats of the 21 species (26 strains) tested. Ethylene, the major volatile produced, was not utilized but butyl acetate, hexyl acetate and hexyl-2-methyl-butanoate (identified by GC-MS) were. These yeasts also assimilated ethanol, butanol, hexanol (Tilletiopsis excepted), acetate, propionate, butyrate and ethyl acetate at appropriately low concentrations. Ethanol and acetate aside, this is the first report of such assimilations by any yeast.

Fruit↗

A quantitative histochemical procedure for measurement of starch in apple fruits.

Measurements of starch (e.g. amyloplasts in stomatal guard cells, sieve elements, root tips or the starch sheath) is often very difficult using most analytical methods. An evaluation was made of interactive computer image analysis of starch measurements in apple fruits. The results obtained indicate that quantitative histochemistry can be an appropriate method to quantify starch. Correlations for starch values between the image analysis system and a colorimetric system were quantified. The thickness of plastic-embedded slices had no influence on the accuracy of the area occupied by image-quantified starch (starch/slice) or on its variance. The magnification of the objective also had no effect on measured starch-occupied areas (starch/slice), but there were big differences in variance. The number of replications required to establish statistically significant differences were calculated.

Histocytochemistry↗

Effect of 2-deoxy-D-glucose on mycotoxins from apples inoculated with Penicillium expansum.

Patulin concentration was not significantly different in Bramley and Cox's apples inoculated with Penicillium expansum, and treated with the biocontrol enhancer 2-deoxy-D-glucose (DOG) when compared to no DOG treatment, despite average numerical increases. Some additional small HPLC peaks were detected from some extracts, one of which corresponded to citrinin.

Journal Article↗

Exposure of California quail to organophosphorus insecticides in apple orchards in the Okanagan Valley, British Columbia.

We studied the exposure and effect of the organophosphate insecticides azinphos-methyl and diazinon on adult California quail (Callipepla californica) in an apple orchard in the Okanagan Valley, British Columbia. Cholinesterase activity was measured in plasma samples (n = 65) collected from 54 individuals either prior to spraying, immediately (< 24 hours) or 10 days after three spray events. Mean plasma cholinesterase levels declined significantly (P < 0.05, n = 12) to 61% of pre-spray mean activity (controls) immediately following the first spray event, but by ten days had recovered to 86% of mean control activity. Subsequent spray events caused no significant declines in mean plasma cholinesterase activity. Four of the 26 quail sampled within 24 h of a spray event exhibited plasma-ChE inhibition exceeding 50% inhibition. Radio-tagged quail (n = 25) were monitored throughout the breeding season to determine use of orchards and detect changes in use patterns resulting from the spraying of insecticides. Use of orchards by quail varied over the summer, with the highest use occurring in May, declining to very low use by July. Quail exhibited a diurnal pattern, roosting in sparsely forested uplands at night, travelling to orchard areas to feed early each morning and returning to roosts at dusk. Orchard use by quail differed during spray events compared to non-spray times. During the three hour period immediately after spraying (0530-0800), 14-20% of observed quail were in the orchard, after which use declined to < 4%, and returned to 12% by the next day. During non-spray times, 3-13% of radio-tagged the quail were observed in orchard habitat, with the heaviest use (13%) occurring later in the day (0830-1700 h). Seven radio-tagged quail were predated during the study period. However, no deaths could be attributed to insecticide poisoning as carcasses were not in suitable condition for testing. It was concluded that adult quail using orchard habitat early in the summer may be acutely poisoned by anti-cholinesterase insecticides, but the risk of exposure declined over the summer.

Agriculture↗

Species association among predaceous and phytophagous apple mites (Acari: Eriophyidae, Phytoseiidae, Stigmaeidae, Tetranychidae).

Predator-predator, predator-prey, and prey-prey associations among nine species of mites were studied in a plot of 100 'Red Delicious' apple (Malus pumila Miller) trees from 1990 to 1997. In 1990, seven-year-old trees were inoculated with Panonychus ulmi (Koch), Tetranychus urticae Koch (Acari: Tetranychidae) or both, and sprayed with azinphosmethyl (alone or plus endosulfan), or nothing. The species Zetzellia mali (Ewing) (Acari: Stigmaeidae), Amblyseius andersoni Chant (Acari: Phytoseiidae), Eotetranychus sp., Bryobia rubrioculus (Scheuten) (Acari: Tetranychidae), and Aculus schlechtendali Nalepa (Acari: Eriophyidae) were already present or immigrated into plots, and Galendromus occidentalis (Nesbitt) and Tvphlodromus pyri Scheuten (Acari: Phytoseiidae) were introduced. Yule's V association index was used to measure positive, neutral, or negative interspecific associations for each species pair, because of its robustness with spatially autocorrelated data. We found that pesticide and release treatments did not greatly affect the association results, but there were strong seasonal differences. Predator-predator associations were the strongest and most consistent, showing negative associations in the early and mid seasons, and neutral ones in late season. Negative associations of T pyri with other predators were the strongest, which is consistent with evidence that this mite can detect other predators on a leaf. Predatorprey seasonal associations were mixed, with some positive and others negative, with most significant associations occurring in the mid season. One prey-prey interaction was positive, again in mid season, most likely because of similar habitat preferences.

Animals↗

Use of apple pomace as a source of dietary fiber in cakes.

Dried and powdered apple pomace was passed through 30, 50 and 60 mesh sieves to get pomace of varying particle size. Blends were prepared by mixing 5, 10 and 15 percent pomace from each of the three particle sizes with wheat flour. The blends were evaluated for cake. Batter viscosity increased with increasing pomace level and decreasing particle size. Specific gravity and pH of the batter decreased with increasing pomace levels. Cake weight, shrinkage and uniformity index increased with increasing pomace levels, whereas, cake volume and symmetry index showed a reverse trend.

Cooking↗

Metabolism of ovorubin, the major egg lipoprotein from the apple snail.

The site of synthesis of molluscs lipoproteins is little known and was investigated for the egg lipoprotein perivitellin 1 (PV 1) or ovorubin in the freshwater snail Pomacea canaliculata. Tissues (albumen gland, gonad-digestive gland complex and muscle) of vitellogenic females were incubated in vitro at 25 degrees C for 12 h with 14C Leucine. After that, soluble proteins from tissue homogenates and medium samples were analysed for de novo protein synthesis by electrophoresis and HPLC, and radiolabelled proteins quantified by liquid scintillation. Gonad-digestive gland complex did not synthesise ovorubin, in spite its high protein synthesis levels. Three albumen gland radiolabelled proteins (35, 32 and 28 kDa) comigrated with the subunits of ovorubin and represented 1.3% of the total labelled protein of that tissue. Western blot analysis with polyclonal antibodies confirmed that these were ovorubin subunits. In vivo experiments where vitellogenic females were injected with 3H Leucine, revealed that ovorubin was not present in hemolymph. ELISA analysis confirmed ovorubin presence only in albumen gland and developing eggs with levels of 800 and 582 mg/g protein, which represent 30.3 and 28.4 mg ovorubin/g of tissue, respectively. Therefore, albumen gland is the single site of ovorubin synthesis as no extragland synthesis, circulation or accumulation could be detected in the apple snail.

Animals↗

Volatiles production and attractiveness to the Mexican fruit fly of Enterobacter agglomerans isolated from apple maggot and Mexican fruit flies.

We investigated two strains of uricase (+) Enterobacter agglomerans, one isolated from the apple maggot fly (AMF) and one from the Mexican fruit fly (MFF), for 1) attractiveness to MFF, and 2) production of attractive chemicals. Regarding chemicals demonstrated attractive to the MFF, the MFF bacterial strain produced more 2,5-dimethylpyrazine, 2-phenylethanol, and indole than the AMF strain, whereas the AMF, but not the MFF strain, produced 3-hydroxybutanone. Cell types that predominated in plated subcultures varied from batch to batch resulting in variation in volatiles production, especially by the AMF strain where indole was sometimes a major component of the odor and at other times not detectable. Despite the greater production of attractive chemicals by the MFF strain, the AMF strain was consistently more attractive and the MFF strain was not different from uninoculated control plates. Statistical analyses indicated negative correlations of attractiveness with production of indole, 2,5-dimethylpyrazine, and 2-phenylethanol, and positive correlation with 3-hydroxybutanone. Results support previous findings with the Mexican fruit fly that showed combinations of attractive chemicals sometimes are not attractive.

Animals↗

Determination of the allergenic activity of birch pollen and apple prick test solutions by measurement of beta-hexosaminidase release from RBL-2H3 cells. Comparison with classical methods in allergen standardization.

BACKGROUND: A murine in vitro model of the allergic type I reaction was set up to determine the biologic activity of extracts without involvement of human beings. It is based on beta-hexosaminidase release from passively sensitized RBL cells after allergen challenge. The intended application of this RBL cell assay in the field of quality control of allergenic extracts requires its comparison with established methods. METHODS: The activity of five standardized birch-pollen prick test solutions was determined in parallel by RBL assay, direct IgE binding, IgE-binding inhibition, major allergen content, histamine-release assay, and skin testing. RESULTS: The RBL cell-release assay corresponded well to other methods if a reagin raised against natural birch-pollen extract was used for passive sensitization. However, in the case of a reagin against recombinant Bet v 1, only a decreased activity was observed, presumably because a reduced number of epitopes were recognized by the monospecific reagin. In contrast to standardized birch-pollen extracts, nonstandardized apple extracts showed poor activity in all assays. CONCLUSIONS: This murine model might be a useful tool in the quality control of allergenic extracts. It combines properties of assays based on standardized antisera and of assays that consider IgE cross-linking properties.

Allergens↗

Membrane-associated protein kinase activities in developing apple fruit.

Fruit development is a process involving various signals and gene expression. Protein phosphorylation catalysed by protein kinases is known to play a key role in eukaryotic cell signalling and so may be involved in the regulation of fruit development. Using the method of exogenous substrate phosphorylation, the activity of calcium-dependent and calmodulin-independent protein kinase (CDPK) that was stimulated by phosphatidylserine, and the myelin basic protein (MBP)-phosphorylating activity that could be due to a calcium-independent mitogen-activated protein kinase-like (MAPK-like) activity in the developing apple fruits were identified. The CDPK activity was shown to be predominantly localized in the plasma membrane, whereas in the presence of phosphatidylserine, the high activity of CDPK was detected in both plasma membrane and endomembranes. The MAPK-like activity was predominantly associated with endomembranes. The assays of bivalent cation requirement showed that Mn2+ could replace Mg2+ in the incubation system for the protein kinase activities and stimulate CDPK activity more than Mg2+. Heat treatment abolished CDPK but stimulated MAPK-like activity. The activities of the phosphatidylserine-stimulated CDPK and of the MAPK-like were fruit developmental stage-specific with higher activities of both enzymes in the early and middle developmental stages in comparison with the late developmental stage. These data suggest that the detected protein kinases may play an important role in the fruit development.

Journal Article↗