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Inactivation of Escherichia coli O157:H7 and other naturally occurring microorganisms in apple cider by electron beam irradiation.

Two Escherichia coli O157:H7 strains, SEA 13 B88 gfp 73ec and B6-914 gfp 90ec, together with two bacteria, three yeasts, and two molds that were randomly selected from a collection of microorganisms found on apples or in apple cider, were inoculated into apple cider and subjected to electron beam irradiation at several doses between 0.0 and 2.3 kGy at the Iowa State University Linear Accelerator Facility. The D-values for the E. coli O157:H7 strains ranged between 0.25 and 0.34 kGy; the D-values for most of the normal flora from apples ranged between 0.24 and 0.59 kGy. By taking into account possible variations in treatment conditions, it was calculated that irradiation at 2.47 kGy should achieve a 5-log reduction of E. coli O157:H7 in apple cider at the 95% confidence level. Naturally occurring yeasts might survive such irradiation treatment.

Beverages↗

Acid tolerance of Escherichia coli O157:H7 and its survival in apple juice.

Outbreaks of diarrhoea and haemolytic uraemic syndrome have been associated with the consumption of apple cider and apple juice. The organism implicated in these outbreaks has been Escherichia coli O157:H7, indicating the resistance of the serotype to acidic pH. On comparing the growth of this serotype with a control strain of E. coli, it was found that strain O157:H7 grew well in trypticase soy broth at pH levels ranging from 2.0 to 9.0, while control strains failed to grow at pH levels below 4.0 and above 9.0. The growth of both strains were inhibited by adding 0.05% of either benzoic acid or sorbic acid. Similarly, O157:H7 grew well in both natural (unpasteurized) as well as in pasteurized apple juice and the growth was inhibited by adding 0.1% of either benzoic acid or sorbic acid. Control strains of E. coli failed to grow in either types of apple juice. The possible sources of contamination of natural apple juice with O157:H7 serotype are discussed.

Acids↗

[Optimum meteorological factors and climate divisions of apple for good quality].

The investigation and laboratory analysis were carried out for apple quality of Fuji, Jonagold and Starkrimson grown in various ecological and climate regions of China. Multiple statistical analysis, linear program and computer technology were used to obtain the optimum values and scheme of major meteorological factors affecting the quality components of different apple varieties. The cluster analysis results showed that it was suitable for apple of Fuji, Jonagold and Starkrimson growing in the Loess Plateau and Longdong, Gansu Province. Bohaiwan bay regions were suitable for apple of Fuji and Jonagold and Southwestern mountainous region were suitable for apple Starkrimson. Advantage qualifications and limiting factors in each climate divisions were also pointed out.

Climate↗

Apples in the American diet.

Apples play an important role in the American diet. They are among the most popular fruits consumed in the United States and are well recognized for their flavor and nutrition. In the last three decades, per capita consumption of processed apples has increased and consumption of fresh apples has declined. Apples are a major food source of flavonoids, antioxidants known for their free radical-scavenging properties. Increasing evidence from in vitro, in vivo, and epidemiologic studies suggest that flavonoids found in apples may be protective against cancer, heart disease, and other chronic diseases.

Diet↗

The effect of a high consumption of apples or grapes on dental caries and periodontal disease in humans.

The intent of this study was to determine the effect of a high consumption of either apples or grapes on the caries experience (DMFT), and periodontal health. Farm workers employed by apple-producing, grape-producing and grain (control) producing farms in low fluoride areas (F less than 0.10 ppm) were investigated. To avoid contamination, only these farms where either apples, grapes or grain were cultivated solely, were included in the study. The caries incidence was found to be the highest for the apple group (24.2), lower for the grape group (17.4), and the lowest for the control group (9.9). The caries incidence differed statistically significantly (P less than 0.01) among the three groups. A higher caries incidence was found in the group of older subjects (35+ years) than in the 15-34 year old group. In general, for both age groups the control group showed conspicuously the highest number of dentate sextants with advanced periodontitis. Significantly less (P less than 0.01) advanced periodontitis and pocket formation could be seen in the youngest age group (15-34 years) of all three test groups. Thus, to conclude, it was found that the consumption of a high amount of apples and to a lesser degree grapes contributed significantly to dental caries. However, the above fruits had a beneficial effect on the periodontal status.

Adolescent↗

Determination of D-malic acid in apple juice by liquid chromatography: collaborative study.

Eleven laboratories collaboratively studied a liquid chromatographic (LC) method for determination of D-malic acid in apple juice. The mobile phase consisted of mM L-valine and 8 mM copper acetate adjusted to pH 5.5 with NaOH. The UV detector was set at 330 nm, and a single reversed-phase LC column was used. Seven paired samples containing various amounts of D-malic acid ranging from 0 to 188 mg/100 mL of 12 Brix pasteurized apple juice were tested by each collaborator. Repeatability and reproducibility coefficients of variation ranged from 1.0 to 3.5% and 7.7 to 11.7%, respectively, within the range of 26 to 188 mg D-malic acid/100 mL of 12 Brix apple juice. The collaborative study results demonstrated that the method could quantitate the economic adulteration of apple juice with DL-malic acid at lower levels than those reported with previous methods. The LC method for determination of D-malic acid in apple juice has been adopted first action by AOAC INTERNATIONAL.

Beverages↗

[New methods for evaluation and analysis of organoleptic qualities of foods and for calculation of changes. 9. Dependence of irradiation flavor of apple juice and concentrate on the gamma radiation dose].

Various authors have defined organoleptically the concept "cooking flavour" for heat-treated apple juice and concentrate. In a similar way, the "irradiation flavour" caused by gamma radiation is defined in the present paper. It has been found that the irradiation flavour is attributable to low-boiling substances which appear already in the first fraction if the modified MICKO distillation is performed. Tetrahydrofuran (THF) has been identified as the key substance in irradiation flavour. As further evidence may be quoted the similarity of the intensity of off-flavour which was observed also by gas chromatography at the same times of retention, and the removal of the irradiation flavour by means of a THF-specific agent (mercury acetate). In connection with THF solutions added to non-irradiated apple juice, the authors employed the technique of "subjective olfactometry", using the "absolute value scale", to establish mathematical correlations between the intensity of irradiation flavour and the concentration of off-flavour products in apple juice or concentrate. The dependence of the intensity of irradiation flavour on the THF concentration and the gamma radiation dose, respectively, obeyed an exponential function. Thus, it has been possible to calculate the amounts of THF formed by radiation doses varying from 0.64 to 2.55 Mrad. Furthermore, the authors determined the minimum radiation dose for 12% apple juice and for 67% apple concentrate at which the irradiation flavour is first perceived (treshold of perception).

Cobalt Radioisotopes↗

Apple juice. An unappreciated cause of chronic diarrhea.

Chronic nonspecific diarrhea (CNSD) remains a common pediatric problem. Previous reports have suggested disordered small intestinal motility, food intolerances, dietary fat restriction, and excessive fluid consumption as possible contributory factors. We have recently encountered a subset of children with CNSD in whom nonexcessive apple juice intake seemed to cause their diarrhea. In five subjects, ingestion of 240 mL of apple juice disclosed evidence of significant carbohydrate malabsorption by breath-hydrogen testing and resulted in diarrhea. Withdrawal of apple juice from the diets of these subjects was curative in all cases. Before embarking on an expensive and time-consuming evaluation for CNSD in otherwise healthy children, a brief restriction of apple juice intake may be warranted.

Breath Tests↗

Apple-peel intestinal atresia associated with balanced reciprocal translocation t(2;3)(q31.3;p24.2) mat.

Apple peel intestinal atresia is an apple-peel-appearing bowel obstruction of unknown cause. We describe a Japanese girl with the apple-peel jejunal atresia associated with apparently balanced reciprocal translocation between chromosomes 2 and 3, t(2;3)(q31. 3;p24.2)mat. The translocation breakpoints in the patient may become candidate regions for the putative gene causing apple-peel atresia. Alternatively, the association of the two abnormalities in the patient is coincidental because her phenotypically normal mother had the same chromosome translocation.

Chromosome Banding↗

Effect of activated charcoal on patulin, fumaric acid and some other properties of apple juice.

In this study, 0, 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 g/l amounts of activated charcoal (AC) were added into apple juice with a patulin content of 62.3 ppb obtained from a well-established manufacturing company. Apple juice samples were then mixed for 0, 5, 10, 20, and 30 min, respectively. Considerable reduction in the patulin and HMF values was found while there is a dramatic improvement in the colour and clearness of apple juice. However, AC did not cause a significant decrease in the fumaric acid level of apple juice. The best result was obtained at 3.0 g/l AC mixed for 5 min. In addition, a negligible reduction in brix and pH values of samples was observed.

Anticarcinogenic Agents↗

Rapid detection of respiring Escherichia coli O157:H7 in apple juice, milk, and ground beef by flow cytometry.

BACKGROUND: Rapid and simple methods to detect viable pathogenic microbes in foods and drinks are required. Flow cytometry was used for the rapid detection of respiring Escherichia coli O157:H7 cells in apple juice, milk, and ground beef. METHODS: CTC (5-cyano-2,3-ditolyl tetrazolium chloride) was used to estimate the respiratory activity of bacteria. Fluorescein isothiocyanate (FITC)-labeled anti-E. coli O157:H7 direct antibody (FA) was used for the specific detection of target cells. Food samples were inoculated with starved E. coli O157:H7 and E. coli K-12 cells, and analyzed by both fluorescent microscopy and flow cytometry after double staining with FA and CTC. RESULTS: Respiring E. coli O157:H7 cells in food samples showed strong fluorescence of both FA (green) and CTC (red); thus, they could be clearly and specifically distinguished from respiring E. coli K-12 or inactive cells. A good correlation was achieved in flow cytometric analysis between the numbers of inoculated viable E. coli O157:H7 and those detected in milk and apple juice. The detection threshold for this flow cytometry for E. coli O157:H7 in milk, apple juice, and ground beef was 10(3) cells/ml (milk and apple juice) or 10(3) cells/g (ground beef) of sample when the total bacterial number in the sample was 10(6) cells/ml. CONCLUSIONS: Respiring E. coli O157:H7 in food samples can be detected specifically within a few hours. Flow cytometry with FA-CTC double staining can be used to examine food contamination with various pathogenic microbes demonstrating physiologic activity through the use of a suitable fluorescent antibody.

Animals↗

Apple flavonoids inhibit growth of HT29 human colon cancer cells and modulate expression of genes involved in the biotransformation of xenobiotics.

Flavonoids from fruits and vegetables probably reduce risks of diseases associated with oxidative stress, including cancer. Apples contain significant amounts of flavonoids with antioxidative potential. The objectives of this study were to investigate such compounds for properties associated with reduction of cancer risks. We report herein that apple flavonoids from an apple extract (AE) inhibit colon cancer cell growth and significantly modulate expression of genes related to xenobiotic metabolism. HT29 cells were treated with AE at concentrations delivering 5-50 microM of one of the major ingredients, phloridzin ("phloridzin-equivalents," Ph.E), to the cell culture medium, with a synthetic flavonoid mixture mimicking the composition of the AE or with 5-100 microM individual flavonoids. HT29 cell growth was inhibited by the complex extract and by the mixture. HT29 cells were treated with nontoxic doses of the AE (30 microM, Ph.E) and after 24 h total RNA was isolated to elucidate patterns of gene expression using a human cDNA-microarray (SuperArray) spotted with 96 genes of drug metabolism. Treatment with AE resulted in an upregulation of several genes (GSTP1, GSSTT2, MGST2, CYCP4F3, CHST5, CHST6, and CHST7) and downregulation of EPHX1, in comparison to the medium controls. The enhanced transcriptional activity of GSTP1 and GSTT2 genes was confirmed with real-time qRT-PCR. On the basis of the pattern of differential gene expression found here, we conclude that apple flavonoids modulate toxicological defense against colon cancer risk factors. In addition to the inhibition of tumor cell proliferation, this could be a mechanism of cancer risk reduction.

Cell Division↗

Polyphenolic apple juice extracts and their major constituents reduce oxidative damage in human colon cell lines.

Apple juice containing high amounts of antioxidative polyphenols might protect the intestine against oxidative cell damage. We investigated the preventive effectiveness of polyphenolic juice extracts of different origins (cider and table apples) in comparison to their major constituents in human colon cell lines (Caco-2, HT29). Parameters studied were (oxidative) DNA damage (Comet assay), glutathione level (photometric kinetic assay), cellular redox status (dichlorofluorescein assay) and antioxidant capacity. The extracts (50-250 microg/mL) modulated DNA damage and redox status in a concentration-dependent manner at 24-h incubation. The pomace extraction technology, applied for juice preparation, and the preferential selection of cider apple varieties influenced the polyphenolic pattern and increased the biological effectiveness of the extracts. The preventive potential of major juice constituents (1-100 microM, 24 h) strongly differed: rutin, epicatechin and caffeic acid clearly reduced (oxidative) DNA damage (Caco-2), chlorogenic acid efficiently decreased cellular reactive oxygen species level (HT29, Caco-2). The aglyca quercetin and phloretin exhibited the highest preventive/antioxidant capacity in all assays. The stability of the compounds inversely correlated with their preventive effectiveness and might contribute to the observed cell specific sensitivities. In conclusion, apple juice extracts distinctly reduce oxidative cell damage in human colon cell lines, an effect, which in part can be accounted for by their major constituents.

Beverages↗

Control of blue mold (Penicillium expansum) by fludioxonil in apples (cv Empire) under controlled atmosphere and cold storage conditions.

A reduced risk fungicide, fludioxonil, was tested for its efficacy against blue mold caused by thiabendazole-resistant and -sensitive Penicillium expansum (Link) Thom in apples under three storage conditions. In a co-treatment, fludioxonil and inoculum were applied together to test the protective activity of the fungicide on wounds that had been aged for 1 or 2 days. The fungicide was also tested for its curative activity in post-inoculation treatment on apples that had been inoculated for 1 or 2 days. Fludioxonil was very effective as co-treatment and as post-inoculation treatment. At a concentration of 300 mg litre(-1), fludioxonil gave complete control of post-harvest blue mold caused by the thiabendazole-resistant and -sensitive P expansum for 105 days in controlled atmosphere (CA) storage at 2 (+/-1) degrees C, for 42 days in common cold storage at 4 (+/-1) degrees C and also in a shelf-life study for 6 days at 20 (+/-1) degrees C. Comparison on the effect of fludioxonil in CA storage and common cold storage showed that higher concentrations of fungicide were needed in cold storage than in CA storage. Fludioxonil at a concentration of 450 mg litre(-1), gave 98 and 92% control of blue mold of apples in the simulated shelf-life studies after CA and common cold storages, respectively. Fludioxonil has a potential to be incorporated in the fungicide resistance management strategies for control of blue mold in apples stored for 105 days.

Cold Temperature↗

Biological activity of carotenoids in red paprika, Valencia orange and Golden delicious apple.

Carotenoid fractions were extracted from red paprika, Valencia orange peel and the peel of Golden delicious apple. Thus, hypophasic carotenoids of paprika (PM1), orange (PM3) and apple (PM4), and epiphasic extractions of paprika (PM2) and apple (PM5) were obtained by extraction, saponification and partition between MeOH-H(2)O (9:1) (hypophasic) and hexane (epiphasic). A high content of capsanthin was quantified in hypophasic carotenoids (PM1) from red spice paprika, whereas the hypophasic fractions from orange (PM3) and apple (PM4) were mainly composed of violaxanthin, zeaxanthin and lutein. On the other hand, a high content of beta,beta-carotene and beta-cryptoxanthin was found in epiphasic fractions (PM2 and PM5). The extracts were studied for their anti-Helicobacter pylori (H. pylori), anti-human immunodeficiency virus (HIV), cytotoxic, multidrug resistance (MDR) reversal and radical scavenging activity. Among five PM extracts and beta,betacarotene, PM4 showed potent anti-H. pylori activity (MIC(50) = 36 microg/mL), comparable to metronidazole (MIC(50) = 45 microg/mL). The extracts were inactive against HIV. PM3 and PM4 showed slightly higher cytotoxic activity against three human tumor cell lines (squamous cell carcinoma HSC-2, HSC-3, submandibular gland carcinoma HSG) and human promyelocytic leukemic HL-60 cells than against three normal human oral cells (gingival fibroblast HGF, pulp cell HPC, periodontal ligament fibroblast HPLF), suggesting a tumor-specific cytotoxic activity. PM1, PM3 and PM4 displayed much higher MDR-reversing activity than (+/-)-verapamil. ESR spectroscopy demonstrated that PM1-5 and beta,beta-carotene produced little or no detectable radical under alkaline conditions and did not scavenge the O(2) (-) produced by the hypoxanthine and xanthine oxidase reaction. On the other hand, PM1 and PM2 scavenged efficiently 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, whereas singlet oxygen was also quenched efficiently by PM5 and PM2. The data suggest the potential importance of carotenoids as possible anti-H. pylori and MDR reversal agents. The active principles in the carotenoid extract might differ, depending upon the types of fruits and vegetables.

Anti-Bacterial Agents↗

Liquid chromatography/mass spectrometric determination of patulin in apple juice using atmospheric pressure photoionization.

This paper describes a comparison between atmospheric pressure chemical ionization (APCI) and the recently introduced atmospheric pressure photoionization (APPI) technique for the liquid chromatography/mass spectrometric (LC/MS) determination of patulin in clear apple juice. A column switching technique for on-line extraction of clear apple juice was developed. The parameters investigated for the optimization of APPI were the ion source parameters fragmentor voltage, capillary voltage, and vaporizer temperature, and also mobile phase composition and flow rate. Furthermore, chemical noise and signal suppression of analyte signals due to sample matrix interference were investigated for both APCI and APPI. The results indicated that APPI provides lower chemical noise and signal suppression in comparison with APCI. The linear range for patulin in apple juice (correlation coefficient >0.999) was 0.2-100 ng mL(-1). Mean recoveries of patulin in three apple juices ranged from 94.5 to 103.2%, and the limit of detection (S/N = 3), repeatability and reproducibility were 1.03-1.50 ng mL(-1), 3.9-5.1% and 7.3-8.2%, respectively. The total analysis time was 10.0 min.

Beverages↗

An ethylene-related cDNA from ripening apples.

We report the isolation of a ripening-related apple cDNA which is complementary to a mRNA which may be involved in ethylene production. Poly(A)+ RNA was extracted from cortical tissue of ripe apple fruit (Malus domestica Borkh cv. Golden Delicious) and a cDNA library constructed in the plasmid vector pSPORT. The library was screened with pTOM13, a tomato cDNA clone thought to code for ACC oxidase in that fruit. An apple cDNA clone (pAP4) was isolated and sequenced. The 1182 bp cDNA insert includes an open reading frame of 942 bp, and shows strong homology with reported tomato and avocado sequences, both at the nucleic acid and amino acid levels. The polypeptide has a calculated molecular mass of 35.4 kDa and a calculated pI of 5.15. In apple cortical tissue, expression of pAP4-complementary RNA increased with ethylene production by the fruit during ripening. Expression was also enhanced in both ethylene-treated and wounded fruit.

Amino Acid Sequence↗

New findings in apple S-genotype analysis resolve previous confusion and request the re-numbering of some S-alleles.

Apple trees display gametophytic self-incompatibility which is controlled by a series of polymorphic S-alleles. To resolve the discrepancies in S-allele assignment that appeared in the literature, we have re-examined the identity of S-alleles known from domestic apple cultivars. Upon an alignment of S-allele nucleotide sequences, we designed allele-specific primer pairs to selectively amplify a single S-allele per reaction. Alternatively, highly similar S-alleles that were co-amplified with the same primer pair were discriminated through their distinct restriction digestion pattern. This is an extension of our previously developed allele-specific PCR amplification approach to reveal the S-genotypes in apple cultivars. Amplification parameters were optimised for the unique detection of the 15 apple S-alleles of which the nucleotide sequences are known. Both the old cultivars with a known S-genotype and a number of more common cultivars were assayed with this method. In most cases, our data coincided with those obtained through phenotypic and S-RNase analysis. However, three S-alleles were shown to relate to RNases that were previously proposed as being encoded by distinct S-alleles. For another S-allele the corresponding gene product has not been discriminated. Consequently, we propose the re-numbering of these four S-alleles. Furthermore, two alleles that were previously identified as S(27a) and S(27b) now received a distinct number, despite their identical S-specificity. To ease widespread future analysis of S-genotypes, we identified common cultivars that may function as a witness for bearing a particular S-allele. We discuss the assignment of new S-alleles which should help to avoid further confusion.

Alleles↗