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PubMed2002

Activity and concentration of polyphenolic antioxidants in apple juice. 1. Effect of existing production methods.

Apples are an important source of flavonoids in the human diet. The effect of processing apples into juice on polyphenolic antioxidant content and activity is described. Raw juice obtained from Jonagold apples by pulping and straight pressing or after pulp enzyming had an antioxidant activity that was only 10 and 3%, respectively, of the activity of the fresh apples. The levels of flavonoids and chlorogenic acid in the juice were reduced to between 50% (chlorogenic acid) and 3% (catechins). Most of the antioxidants were retained in the pomace rather than being transferred into the juice. Apparently, most of the antioxidant compounds are absorbed to the solid matter of the pomace. In apple juice, 45% of the total measured antioxidant activity could be ascribed to the analyzed antioxidants. For three apple cultivars tested (Elstar, Golden Delicious, and Jonagold), the processing methods had similar effects. The results indicate that processing can have a major impact on the bioactivity of products.

Antioxidants↗
PubMed2004

Technological classification of basque cider apple cultivars according to their polyphenolic profiles by pattern recognition analysis.

The polyphenolic compositions of 31 Basque cider apple cultivars were determined in pulp, peel, and juice by high-performance liquid chromatography--diode array detection analysis of crude extracts and after thiolysis. Data sets, consisting of individual polyphenol concentrations, total procyanidin content, and the average degree of polymerization of procyanidins, were evaluated by multivariate chemometric techniques, to develop decision rules for classifying apple cultivars technologically into bitter and nonbitter categories. A preliminary study of the data structure was performed by cluster analysis and principal component analysis in each apple material. Bitter apple varieties presented higher contents of flavan-3-ols and/or dihydrochalcones than nonbitter cultivars. Different classification systems for the two categories on the basis of the chemical data were obtained applying several supervised pattern recognition procedures, such as linear discriminant analysis, K-nearest neighbors, soft independent modeling of class analogy, partial least-squares, and multilayer feed forward artificial neural networks. Excellent performance in terms of recognition and prediction abilities for both categories (100% of hits) was achieved in every case (pulp, peel, or juice). Polyphenolic profiles of apple pulp, peel, or juice provide enough information to develop classification criteria for establishing the technological group of apple cultivars (bitter or nonbitter).

Analysis of Variance↗
PubMed2004

Ester variability in apple varieties as determined by solid-phase microextraction and gas chromatography-mass spectrometry.

Solid-phase microextraction (SPME) with a polydimethylsiloxane fiber coupled with gas chromatography-mass spectrometry (GC-MS) was applied to the study of variability in volatiles released by 13 apple varieties. The relative amounts of 40 esters and alpha-farnesene were determined. Principal component analyses of these results clustered the apples into three groups according to skin color: red, green, and red-green. Total ester contents were highest with the red cluster apples, and the green cluster apples had the highest alpha-farnesene levels. This technology was also applied to the monitoring of changes in volatiles for apples removed from controlled-atmosphere storage with subsequent storage at 4 degrees C and room temperature. Total ester contents increased 25-fold, with the greater increases coming at room temperature, whereas alpha-farnesene levels increased only 5-fold. For apples stored at room temperature, after 11 days, the amount of increase was inversely proportional to the size of the ester: levels of smallest esters (molecular weight 116) increased 12.5-fold, and the largest esters (molecular weight 228) increased approximately 1.3-fold.

Esters↗
PubMed2006

Maturity and storage influence on the apple (Malus domestica) allergen Mal d 3, a nonspecific lipid transfer protein.

Consumption of apples can provoke severe allergic reactions, in susceptible individuals, due to the presence of the allergen Mal d 3, a nonspecific lipid transfer protein, found largely in the fruit skin. Levels of Mal d 3 were determined in peel as a function of apple cultivar, position of the fruit growing on the tree, apple maturity, and postharvest storage by ELISA. As the apples mature, Mal d 3 levels increased, although the rate was dependent on cultivar and tree position. During storage, levels of Mal d 3 decreased in all cultivars (cvs. Cox, Jonagored, and Gala), the rate of overall decrease being greatest under controlled atmosphere conditions. There was no correlation between Mal d 3 levels and total apple peel protein, indicating specific alterations in Mal d 3 expression. Thus pre- and postharvest treatments (i.e., storage) can modify the allergen load in apple peel, the highest levels being found in overly mature and freshly harvested fruits.

Allergens↗
PubMed1984

Calmodulin inhibitor in senescing apples and its physiological and pharmacological significance.

While assaying calmodulin activity in senesced apple extracts by using its property of promoting the activity of activator-deficient 3',5'-cyclic AMP 5'-nucleotidohydrolase (phosphodiesterase, EC 3.1.4.17) from bovine heart, we detected a heat-stable, dialyzable, low molecular weight component that inhibited calmodulin activity. Specific activity of calmodulin as calculated from the linearly increasing portion of the activity curve was in the range of 150 to 160 units/mg of protein in crude extracts from apples stored at 2 degrees C for a period of 6 months with or without calcium treatment. Apple extract that was passed through a phenothiazine-Sepharose affinity column did not promote phosphodiesterase activity, whereas the EGTA eluate of the column promoted phosphodiesterase activity similar to the original extract. The inhibition of calmodulin activity appeared to be lower in extracts from apples stored at 2 degrees C after calcium treatment. The inhibition was found to increase after storage of apples at room temperature for 30 days. Activity of purified bovine brain calmodulin was also inhibited by the inhibitor present in apple extracts, which indicated that the inhibition was not specific to plant calmodulin alone and could have wide applications. The importance of the inhibitor in relation to senescence/aging and its possible pharmacological applications are discussed.

Journal Article↗
PubMed2003

Inhibition of AP-1 and neoplastic transformation by fresh apple peel extract.

Consumption of fruits and vegetables has been associated with a low incidence of cancers and other chronic diseases. Previous studies suggested that fresh apples inhibit tumor cell proliferation. Here we report that oral administration of apple peel extracts decreased the number of nonmalignant and malignant skin tumors per mouse induced by 12-O-tetradecanolyphorbol-13-acetate (TPA) in 7,12-dimethylbenz(a)anthracene-initiated mouse skin. ESR analysis indicated that apple extract strongly scavenged hydroxyl (OH) and superoxide (O(2)(-)) radicals. Mechanistic studies showed that pretreatment with apple peel extract inhibited AP-1 transactivation induced by ultraviolet B irradiation or TPA in JB6 cells and AP-1-luciferase reporter transgenic mice. This inhibitory effect appears to be mediated by the inhibition of ERKs and JNK activity. The results provide the first evidence that an extract from fresh apple peel extract may inhibit tumor promoter-induced carcinogenesis and associated cell signaling, and suggest that the chemopreventive effects of fresh apple may be through its antioxidant properties by blocking reactive oxygen species-mediated AP-1-MAPK activation.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed2003

Effects of storage conditions and fruit processing on the degradation of parathion methyl on apples and lemons.

Degradation of parathion methyl on lemon and apple on the trees, during storage of these fruits in refrigerated rooms, and during juice production was studied. The pesticide was applied to the trees according to the recommended application procedures. Apples and lemons received a single application of parathion methyl at 40 g active ingredient/100 litres. Residues were determined with a simple gas-chromatographic method; the recovery rate of parathion methyl from apples and lemons was 85-108% and the limit of determination was 0.004 mg kg(-1). The half-lives of parathion methyl on the trees were 7 days for apples and 5 days for lemons. The duration of storage of the fruits in a refrigerated room have been largely extended to 65 and 63 days, respectively. The high acidity of lemons was found not to affect the degradation rate of parathion methyl. When fruit juice was produced from both apples and lemons, parathion methyl residues were detected in apple but not in lemon juice.

Citrus↗
PubMedDate not supplied

Quantifying the 'appleness' or 'pearness' of the human body by subcutaneous adipose tissue distribution.

To quantify subcutaneous adipose tissue topography (SAT-Top) describing individual SAT distribution for a subject or even a group we measured subcutaneous adipose tissue thickness at 15 specified body sites of 303 healthy women aged 20-69 yrs and 20 women with proven non-insulin-dependent diabetes mellitus (NIDDM) by the optical device 'LIPOMETER'. The type of upper-body- and lower-body-fat pattern, (apples or pears), was determined by factor analysis of the data. Upper body sites were highly loaded in factor 1, whereas factor 2 included highly loaded body sites from the lower extremities. For an individual, factor 1 scores > factor 2 scores, was described as an 'apple'-type, while factor 2 > factor 1 was described as a 'pear'-type. We found about 80% 'pears' and 20% 'apples' in 20-29 year olds and 20% 'pears' and 80% 'apples' in 60-69 year old women. Women with NIDDM tended to be 'super-apples'. SAT-Top provides a useful differentiation between apples and pears and we recommend this approach as a screening method.

Adipose Tissue↗
PubMed2003

Evidence for inversion polymorphism related to sympatric host race formation in the apple maggot fly, Rhagoletis pomonella.

Evidence suggests that the apple maggot, Rhagoletis pomonella (Diptera: Tephritidae) is undergoing sympatric speciation (i.e., divergence without geographic isolation) in the process of shifting and adapting to a new host plant. Prior to the introduction of cultivated apples (Malus pumila) in North America, R. pomonella infested the fruit of native hawthorns (Crataegus spp.). However, sometime in the mid-1800s the fly formed a sympatric race on apple. The recently derived apple-infesting race shows consistent allele frequency differences from the hawthorn host race for six allozyme loci mapping to three different chromosomes. Alleles at all six of these allozymes correlate with the timing of adult eclosion, an event dependent on the duration of the overwintering pupal diapause. This timing difference differentially adapts the univoltine fly races to an approximately 3- to 4-week difference in the peak fruiting times of apple and hawthorn trees, partially reproductively isolating the host races. Here, we report finding substantial gametic disequilibrium among allozyme and complementary DNA (cDNA) markers encompassing the three chromosomal regions differentiating apple and hawthorn flies. The regions of disequilibrium extend well beyond the previously characterized six allozyme loci, covering substantial portions of chromosomes 1, 2, and 3 (haploid n = 6 in R. pomonella). Moreover, significant recombination heterogeneity and variation in gene order were observed among single-pair crosses for each of the three genomic regions, implying the existence of inversion polymorphism. We therefore have evidence that genes affecting diapause traits involved in host race formation reside within large complexes of rearranged genes. We explore whether these genomic regions (inversions) constitute coadapted gene complexes and discuss the implications of our findings for sympatric speciation in Rhagoletis.

Adaptation, Physiological↗
PubMed2005

Evidence of sexual reproduction of woolly apple aphid, Eriosoma lanigerum, in New Zealand.

Reproduction of the woolly apple aphid, Eriosoma lanigerum (Hausmann), can take place parthenogenetically or sexually when both host plants, apple (Malus domestica Borkh.) and elm (Ulmus americana L.) are available. Since elm is not commonly grown in New Zealand, E. lanigerum, a major pest of apple, is thought to reproduce only parthenogenetically here. During our studies between 1999 and 2003, different morphs of E. lanigerum have been observed on apple trees, which were studied in more detail in 2003 and 2004. In the laboratory, alates reproduced mainly sexual morphs with degenerated mouthparts. Oviparous females lived for 8.95 +/- 0.17 days (n = 20) and males lived for 7.1 +/- 0.12 days (n = 20). Both went through 4 moults, without feeding or changing body size. Oviparous female laid a single egg and died soon after oviposition. In addition to sexual morphs, shiny brown, oblong eggs were seen on apple leaves grown outside as well as in the glasshouse. As the numbers of eggs and sexual morphs on trees grown outside were less than on those grown in the glasshouse, we suggest that alates disperse into the natural environment searching for an apple or elm tree to continue the sexual reproduction while spreading the population.

Animals↗
PubMed2006

Light-induced expression of a MYB gene regulates anthocyanin biosynthesis in red apples.

Anthocyanins are secondary metabolites found in higher plants that contribute to the colors of flowers and fruits. In apples (Malus domestica Borkh.), several steps of the anthocyanin pathway are coordinately regulated, suggesting control by common transcription factors. A gene encoding an R2R3 MYB transcription factor was isolated from apple (cv Cripps' Pink) and designated MdMYB1. Analysis of the deduced amino acid sequence suggests that this gene encodes an ortholog of anthocyanin regulators in other plants. The expression of MdMYB1 in both Arabidopsis (Arabidopsis thaliana) plants and cultured grape cells induced the ectopic synthesis of anthocyanin. In the grape (Vitis vinifera) cells MdMYB1 stimulated transcription from the promoters of two apple genes encoding anthocyanin biosynthetic enzymes. In ripening apple fruit the transcription of MdMYB1 was correlated with anthocyanin synthesis in red skin sectors of fruit. When dark-grown fruit were exposed to sunlight, MdMYB1 transcript levels increased over several days, correlating with anthocyanin synthesis in the skin. MdMYB1 gene transcripts were more abundant in red skin apple cultivars compared to non-red skin cultivars. Several polymorphisms were identified in the promoter of MdMYB1. A derived cleaved amplified polymorphic sequence marker designed to one of these polymorphisms segregated with the inheritance of skin color in progeny from a cross of an unnamed red skin selection (a sibling of Cripps' Pink) and the non-red skin cultivar Golden Delicious. We conclude that MdMYB1 coordinately regulates genes in the anthocyanin pathway and the expression level of this regulator is the genetic basis for apple skin color.

Amino Acid Sequence↗
PubMed1983

Allergy to apple, carrot and potato in children with birch pollen allergy.

Skin sensitivity to apple, carrot and potato, clinically related to birch pollinosis was investigated. Different skin test techniques using fresh fruit were compared. A simple prick test (SPT) technique with a lancet piercing the apple peel just before pricking the skin was shown to be the most practical and to give reproducible results. The allergenic activity in apple was found to be heat labile and deteriorated during storage at room temperature. Apple, carrot, potato, hazelnut and birch reactivity was transferable in Prausnitz-Küstner test like IgE antibodies. SPT reactivity to fresh material from apple, carrot and potato was investigated in 174 children of whom 128 suffered from pollen allergy. Positive SPT results were obtained almost exclusively in children who were SPT positive to a birch pollen extract. Children who noticed clinical symptoms when eating apple, raw carrot or potato were found to have a significantly larger SPT reaction than children with a negative history.

Adolescent↗
PubMed2004

Acid resistance systems required for survival of Escherichia coli O157:H7 in the bovine gastrointestinal tract and in apple cider are different.

Escherichia coli O157:H7 is a highly acid-resistant food-borne pathogen that survives in the bovine and human gastrointestinal tracts and in acidic foods such as apple cider. This property is thought to contribute to the low infectious dose of the organism. Three acid resistance (AR) systems are expressed in stationary-phase cells. AR system 1 is sigma(S) dependent, while AR systems 2 and 3 are glutamate and arginine dependent, respectively. In this study, we sought to determine which AR systems are important for survival in acidic foods and which are required for survival in the bovine intestinal tract. Wild-type and mutant E. coli O157:H7 strains deficient in AR system 1, 2, or 3 were challenged with apple cider and inoculated into calves. Wild-type cells, adapted at pH 5.5 in the absence of glucose (AR system 1 induced), survived well in apple cider. Conversely, the mutant deficient in AR system 1, shown previously to survive poorly in calves, was susceptible to apple cider (pH 3.5), and this sensitivity was shown to be caused by low pH. Interestingly, the AR system 2-deficient mutant survived in apple cider at high levels, but its shedding from calves was significantly decreased compared to that of wild-type cells. AR system 3-deficient cells survived well in both apple cider and calves. Taken together, these results indicate that E. coli O157:H7 utilizes different acid resistance systems based on the type of acidic environment encountered.

Animals↗
PubMed2005

Identification by genome scanning approach (GSA) of a microsatellite tightly associated with the apple scab resistance gene Vm.

For all known major apple scab resistance genes except Vr, molecular markers have been published. However, the precise position of some of these genes, in the apple genome, remains to be identified. Knowledge about the relative position of apple scab resistance genes is necessary to preliminarily evaluate the probability of success of their pyramidization. Pyramidization of different resistance genes into the same genotype is a reliable way to create cultivars with durable apple scab resistance. Applying the genome scanning approach (GSA), we identified the linkage group of the scab resistance gene Vm, derived from Malus micromalus, and we found a new molecular marker tightly associated with the gene. The simple sequence repeat Hi07h02, previously mapped on linkage group 17, cosegregates with the Vm gene (no recombinants in the 95 plants tested). The already published sequence-characterized amplified region Vm marker OPB12(687) was found to be linked at about 5 cM from the resistance gene and, therefore, this marker also maps on linkage group 17 of apple. This is the first report of the discovery of a major apple scab resistance gene on linkage group 17. The advantages of using GSA for the identification of molecular markers for qualitative traits are discussed.

Alleles↗
PubMed2006

Effect of apple extracts on NF-kappaB activation in human umbilical vein endothelial cells.

The mechanisms by which foods, such as fruit, are able to reduce the risk of chronic disease are still unclear. Several fruit products, including apples and apple juice, that are flavonoid-rich are reported to increase antioxidant levels in human subjects. This is supported by the finding from our previous studies that the chronic consumption of apple juice by human subjects reduced ex vivo low-density lipoprotein (LDL) oxidation; we hypothesized that this was due to the flavonoid in the apple juice, which, as we reported earlier, reduced in vitro LDL oxidation. To further explore whether the mixture of flavonoids and other phytochemicals in apples are biologically relevant antioxidants, we tested the effects of this flavonoid-rich apple extract (AE) on oxidant-related pathways in a model of the endothelium: human umbilical vascular endothelial cells (HU-VECs). The effects of AE on oxidant-responsive (i.e., tumor necrosis factor [TNF]-alpha-induced) nuclear factor (NF)- kappaB signaling in cell culture were assessed in transfected HUVECs by using a construct that expressed luciferase under the control of NF-kappaB. Incubation of HUVEC for 24 hrs with up to 10 mM (as gallic acid equivalents) of AE demonstrated no cytotoxicity, as determined by lactate dehydrogenase release, caspase 3 activation, and apoptosis marker-based FACS analysis. AE after a 24-hr incubation period at either 200 or 2000 nM showed a complex pattern of decreased basal and TNF-alpha-stimulated NF-kappaB signaling (63% maximal decrease) as assessed by luciferase activity in the transfected HUVECs, as well as by reduced levels of IkappaBalpha protein phosphorylation detected by Western blot analysis. We suggest that AE downregulates NF-kappaB signaling and that this is indicative of an antioxidant effect of the flavonoids present in AE.

Cell Line↗
PubMed2005

Detection of fecal contamination on apples with nanosecond-scale time-resolved imaging of laser-induced fluorescence.

Detection of apples contaminated with feces is a public health concern. We found that time-resolved imaging of apples artificially contaminated with feces allowed optimization of timing parameters for detection. Dairy feces were applied to Red Delicious and Golden Delicious apples. Laser-induced fluorescence responses were imaged by use of a gated intensified camera. We developed algorithms to automatically detect contamination iteratively by using one half of the apples and validated them by applying the optimized algorithms to the remaining apples. Results show that consideration of the timing of fluorescence responses to pulsed-laser excitation can enhance detection of feces on apples.

Algorithms↗
PubMed1996

Effects of different types of dietary fiber preparations isolated from bamboo shoots, edible burdock, apple and corn on fecal steroid profiles of rats.

This study was conducted to evaluate the effects of different types of dietary fibers (DF) under the conditions with or without cholesterol (Chol) loading on the amount and composition of steroids in rat feces. Rats were fed Chol-unsupplied diets containing 10% lard and 5% DF preparation isolated from four kinds of food, bamboo shoots, edible burdock, apple and corn, for three weeks. The respective diets were supplemented with 0.5% Chol and then given to the rats for a further two weeks. The excretion of total bile acid (BA) and several major BAs increased significantly in the apple group with or without Chol loading when compared with that in the cellulose (CP) or other DF groups. The tendency in the apple group was more noticeable when the diet was supplemented with Chol. This is presumably a major reason for the tendency of decrease in serum and liver Chol concentrations in the apple group. The ratio of secondary BAs to total BA in the feces was significantly low in the apple group. Although the lithocholic acid (LCA)/deoxycholic acid (DCA) ratio, a risk index for colorectal cancer, was significantly lower in the bamboo, burdock and apple groups than in the CP or corn groups when given the diet without Chol, the differences disappeared with the addition of Chol. The proportion of coprostanol, a secondary metabolite of Chol, was smaller in the former three groups than in the CP or corn groups. These results suggest that the intake of some DF by host animals works beneficially for the microbial conversion of BA and Chol in the large intestine but that the addition of Chol acts to cancel such beneficial effects.

Animal Feed↗
PubMed1998

Survival of Escherichia coli O157:H7 in synthetic gastric fluid after cold and acid habituation in apple juice or trypticase soy broth acidified with hydrochloric acid or organic acids.

Extreme acid tolerance of Escherichia coli O157:H7 has raised doubts about the safety of acidic foods. This study examined whether prior storage in acidic and/or cold conditions enhanced survival of E. coli O157:H7 in synthetic gastric fluid (SGF). Three E. coli O157:H7 strains were stored in trypticase soy broth (TSB; acidified with HCl, malic acid, citric acid, or lactic acid) or pH 3.5 and 6.5 (nonacidic control) apple juice at 4 and 21 degrees C for < or = 7 days and then were incubated in pH 2.5 SGF at 37 degrees C for 4 h. Cells survived better in apple juice than in TSB containing organic acids, suggesting that juice constituents other than organic acids protect E. coli O157:H7. Refrigeration combined with low pH best protected cells in apple juice and acidified TSB, but, compared to the nonacidic control, only acidified TSB enhanced subsequent survival in pH 2.5 SGF. Equal survival in SGF occurred after storage in pH 3.5 or 6.5 apple juice at 4 degrees C, suggesting that low temperature alone in apple juice enhanced acid tolerance. Two strains stored at 4 degrees C in TSB containing malic or citric acid subsequently survived better in SGF than cells stored in nonacidified TSB but poorer than cells stored in the presence of HCl. These differences reflect the higher pKa of these organic acids. However, subsequent survival of these strains in SGF was poorer after refrigerated storage in apple juice than in TSB containing citric or malic acids. Cells stored in lactic acid were most likely to be completely eliminated upon transfer to SGF. Differences in survival in storage media or SGF related to strain, storage conditions, or acidifier were consistent and often statistically significant (P < 0.05). Although the survival of E. coli O157:H7 in refrigerated acidic beverages may not be affected by the type of acidifier used, the subsequent survival in SGF of this pathogen may be critically dependent on this factor.

Beverages↗