An 'apples-to-apples' comparison of training hours for DOs, PAs, NPs.
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Apples are a major source of flavonoids in the Western diet, and flavonoid-rich foods may help protect against chronic diseases by antioxidant mechanisms. In the present study we investigated: (1) the antioxidant capacity of representative apple polyphenols and their contribution to the total antioxidant capacity of apple extracts; (2) the effects of adding apple extract to human plasma in vitro on oxidation of endogenous antioxidants and lipids; and (3) the effects of apple consumption by humans on ex vivo oxidation of plasma antioxidants and lipids. We found that the apple-contained flavonols and flavanols, quercetin, rutin, (-)-epicatechin, and (+)-catechin, had a higher antioxidant capacity than the dihydrochalcones, phloridzin and phloretin, and the hydroxycinnamate, chlorogenic acid. However, together these apple polyphenols contributed less than 20% to the total antioxidant capacity of aqueous apple extracts. When human plasma was exposed to a constant flux of aqueous peroxyl radicals, endogenous ascorbate (70.0 +/- 10.3 microM) was oxidized within 45 min of incubation, while endogenous urate (375 +/- 40 microM) and alpha-tocopherol (24.7 +/- 1.2 microM) were oxidized after ascorbate. Addition of 7.1 or 14.3 micrograms/ml total phenols of apple extract did not protect ascorbate from oxidation, but increased the half-life (t1/2) of urate from 136 +/- 15 to 192 +/- 16 and 208 +/- 23 min, respectively (p < 0.05 each), and t1/2 of alpha-tocopherol from 141 +/- 18 to 164 +/- 8 min (p = ns) and 188 +/- 8 min (p < 0.05). Lipid peroxidation started after ascorbate depletion, and addition of apple extract increased the lag time preceding detectable lipid peroxidation from 36.3 +/- 3.7 to 50.9 +/- 2.7 min (p < 0.05) and 70.4 +/- 4.2 min (p < 0.001). However, when six healthy volunteers ate five apples and plasma was obtained up to 4 h after apple consumption, no significant increases in the resistance to oxidation of endogenous urate, alpha-tocopherol, and lipids were found. Thus, despite the high antioxidant capacity of individual apple polyphenols and apple extracts and the significant antioxidant effects of apple extract added to human plasma in vitro, ingestion of large amounts of apples by humans does not appear to result in equivalent in vivo antioxidant effects of apple polyphenols.
The Drosophila Appl gene encodes a transmembrane protein that is expressed exclusively in neurons. Amino acid comparisons show that APPL protein is a member of the amyloid precursor protein (APP)-like family of proteins. Similar to mammalian APP-family proteins, APPL is synthesized as a transmembrane holoprotein and cleaved to release a large secreted amino-terminal domain. Using immunocytochemical methods, we have analyzed the distribution of APPL in the Drosophila CNS. Surprisingly, although APPL is present in all neuronal cell bodies, the neurophil shows sterotypic differential distribution. Double-labeling experiments with different neuronal markers were used to distinguish between APPL associated with neuronal processes or extracellular matrix. The distribution of APPL protein produced from transgenes encoding wild-type (APPL), secretion-defective (APPLsd), and constitutively secreted (APPLs) forms was analyzed in an Appl-deficient background to determine which APPL form is associated with different neuropil regions. We found that APPLsd protein is enriched where APPL immunoreactivity coincides with neuronal processes. In contrast, APPLs preferentially localizes to those parts of the neuropil that show a diffuse APPL signal that rarely colocalizes with processes, and thus seems to be a component of the extracellular matrix. These data indicate that proteolytic cleavage and trafficking of APPL is differentially regulated in different neuronal populations. Through metamorphosis, APPL is especially abundant in growing axons and in areas where synapses are forming. Interestingly, in adult brains, APPL protein is enriched in the mushroom bodies and to a lesser extent in the central complex, structures involved in learning and memory.
Patients with tree pollinosis frequently report allergic reactions after ingestion of apples. The severity of apple allergy has been related to the variety of apples and their degree of maturity. To generate a serum pool that is representative of various IgE-binding patterns of apple-allergic sera, serum samples from 34 patients allergic to tree pollens were screened. Only 24 serum samples reacted to the apple extract. Pooled serum was used to identify allergens in apples. An efficient and consistent extraction method for apple fruits was used to compare the immunoreactivities of extracts of different varieties (McIntosh, Red Delicious, Granny Smith, and Golden Delicious) of freshly picked and store-purchased apples. We found that Golden Delicious apples had the greatest amount of the 18 kd allergen, which has been reported to be a potent IgE-binding apple allergen. Store-purchased apples contained higher concentrations of the 18 kd allergen than freshly picked apples. In our study only 37.5% of sera reacted to the 18 kd protein, whereas 75% of the sera reacted to a 31 kd allergen. Other immunoreactive bands in apple extracts included proteins of 50, 38, 16, 14, and 13 kd. The amino-terminal amino acid sequences of the two major allergens, 18 kd and 31 kd, were determined. These sequences shared approximately 50% identity with disease resistance proteins of various plants or Bet v 1 in birch tree pollens. The appearance of various allergens was also investigated in mature apples during storage. The amount of 18 kd allergen increased significantly when apples were stored at 4 degrees C. However, under controlled atmospheric conditions in which oxygen- and carbon dioxide-induced ripening were regulated, the amount of 18 kd allergen remained unaffected. Because ripening and maturation were not associated with increases in 18 kd allergen content, the observed changes might be induced by factors related to disease resistance.
The objectives of this study were (i) to evaluate the survival of coliforms, Escherichia coli, and enterococci in refrigerated apple cider; (ii) to develop simple and inexpensive presumptive methods for detection of these bacteria; (iii) to perform a field survey to determine the prevalence of these bacteria on apples and in apple cider; and (iv) based on our results, to recommend the most useful of these three indicator groups for use in verifying apple cider processing plant sanitation and hazard analysis critical control point (HACCP) programs. Eight of 10 coliform strains (5 E. coli, 1 Enterobacter aerogenes, and 2 Klebsiella spp.) inoculated into preservative-free apple cider (pH 3.4, 13.3(o) Brix) survived well at 4 degrees C for 6 days (< or = 3.0 log10 CFU/ml decrease). Of 21 enterococci strains (Enterococcus faecalis, E. faecium, and E. durans), only 2 E. durans and 3 E. faecium strains survived well. Simple broth-based colorimetric methods were developed that detected the presence of approximately 10 cells of coliforms or enterococci. In three field studies, samples of unwashed apples (drops and picked), washed apples, and freshly pressed cider were presumptively analyzed for total coliforms, E. coli, and enterococci using qualitative and/or quantitative methods. Drop apples were more likely than picked apples to be contaminated with E. coli (26.7% vs. 0%) and enterococci (20% vs. 0%). Washing had little effect on coliform populations and in one field study was associated with increased numbers. Total coliform populations in cider ranged from < 1 CFU/ml to > 738 most probable number/ml, depending on the enumeration method used and the sample origin. E. coli was not recovered from washed apples or cider, but enterococci were present on 13% of washed apple samples. The qualitative coliform method successfully detected these bacteria on apples and in cider. Based on its exclusively fecal origin, good survival in apple cider, and association with drop apples, we conclude that E. coli is the most useful organism for verifying apple cider sanitation and HACCP programs.
There is some evidence that chronic diseases, such as cancer and cardiovascular disease, may occur as a result of oxidative stress. Apple peels have high concentrations of phenolic compounds and may assist in the prevention of chronic diseases. Millions of pounds of waste apple peels are generated in the production of applesauce and canned apples in New York State each year. We proposed that a valuable food ingredient could be made using the peels of these apples if they could be dried and ground to a powder without large losses of phytochemicals. Rome Beauty apple peels were treated with citric acid dips, ascorbic acid dips, and blanches before being oven-dried at 60 degrees C. Only blanching treatments greatly preserved the phenolic compounds, and peels blanched for 10 s had the highest total phenolic content. Rome Beauty apple peels were then blanched for 10 s and dried under various conditions (oven-dried at 40, 60, or 80 degrees C, air-dried, or freeze-dried). The air-dried and freeze-dried apple peels had the highest total phenolic, flavonoid, and anthocyanin contents. On a fresh weight basis, the total phenolic and flavonoid contents of these samples were similar to those of the fresh apple peels. Freeze-dried peels had a lower water activity than air-dried peels on a fresh weight basis. The optimal processing conditions for the ingredient were blanching for 10s and freeze-drying. The process was scaled up, and the apple peel powder ingredient was characterized. The total phenolic content was 3342 +/- 12 mg gallic acid equivalents/100 g dried peels, the flavonoid content was 2299 +/- 52 mg catechin equivalents/100 g dried peels, and the anthocyanin content was 169.7 +/- 1.6 mg cyanidin 3-glucoside equivalents/100 g dried peels. These phytochemical contents were a significantly higher than those of the fresh apple peels if calculated on a fresh weight basis (p < 0.05). The apple peel powder had a total antioxidant activity of 1251 +/- 56 micromol vitamin C equivalents/g, similar to fresh Rome Beauty peels on a fresh weight basis (p > 0.05). One gram of powder had an antioxidant activity equivalent to 220 mg of vitamin C. The freeze-dried apple peels also had a strong antiproliferative effect on HepG(2) liver cancer cells with a median effective dose (EC(50)) of 1.88 +/- 0.01 mg/mL. This was lower than the EC(50) exhibited by the fresh apple peels (p < 0.05). Apple peel powder may be used in a various food products to add phytochemicals and promote good health.
The aim of the study was to develop and evaluate different methods of double-blind, placebo-controlled food challenge (DBPCFC) with apple. Three different DBPCFC models were evaluated: fresh apple juice, freshly grated apple, and freeze-dried apple powder. All challenges were performed outside the pollen season and took place from 1997 to 1999. The freeze-dried apple material was characterized by means of leukocyte histamine release (HR), skin prick test (SPT), and immunoblotting experiments. The study population consisted of birch pollen-allergic patients with a history of rhinitis in the birch-pollen season and positive specific IgE to birch. For comparison of the DBPCFC models, 65 patients with a positive open oral challenge with apple were selected. In the characterization of the freeze-dried apple material, 46 birch pollen-allergic patients were included. The IgE reactivity to apple was evaluated by measurement of specific IgE, HR, and SPT. Golden Delicious apples were used in all experiments. The results of this study showed that it was possible to perform DBPCFC with apple in birch pollen-allergic individuals. The model with freshly squeezed apple juice had a low sensitivity and displayed a high frequency of reactions to placebo, probably due to the ingredients used for blinding. The sensitivity of the models with freshly grated apple and freeze-dried apple powder was 0.74/0.60. An increase in sensitivity is desirable. The freeze-dried apple powder proved to be useful for SPT, HR, and oral challenges, but further investigation of the stability and the allergenic profile of the material is needed.
Dietary factors have been shown to contribute to the occurrence or persistence of chronic non-specific diarrhoea (CNSD). Among these are low dietary fat, high fluid consumption, and the consumption of apple juice. Prompted by the clinical impression that freshly pressed and unprocessed ('cloudy') apple juice was less likely to induce diarrhoea than normal, enzymatically processed ('clear') apple juice, both juices were compared in terms of carbohydrate malabsorption, gastric emptying, and effects on defecation patterns. Clear and cloudy apple juice differ in their fibre and non-absorbable monosaccharide and oligosaccharide contents. Ten healthy children aged 3.6 to 5.9 years ingested 10 ml/kg of clear and cloudy apple juice; in five of them it was enriched with 40 mg of [1-13C]-glycine. Clear apple juice resulted in increased (> or = 20 ppm) breath hydrogen excretion in 8/10, compared with 5/10 after cloudy apple juice; peak breath hydrogen was higher in the clear apple juice group (35 (4) and 18 (3) ppm, respectively). Gastric emptying as determined by means of labelled breath carbon dioxide (13CO2) excretion was similar with both juices. In a four week crossover clinical trial 12 children, formerly diagnosed as having CNSD, were given extra clear fluids (excluding fruit juices; > or = 50% over basal consumption), clear apple juice, or cloudy apple juice, for five day periods. Extra fluids and cloudy apple juice did not influence stool frequency and consistency compared with the basal period. In contrast, clear apple juice significantly promoted diarrhoea. It is suggested that, in addition to fructose, the increased availability of non-absorbable monosaccharides and oligosaccharides as a result of the enzymatic processing of apple pulp is an important aetiological factor in apple juice induced CNSD.
Consumption of fruits and vegetables has been shown to be effective in the prevention of chronic diseases. These benefits are often attributed to the high antioxidant content of some plant foods. Apples are commonly eaten and are large contributors of phenolic compounds in European and North American diets. The peels of apples, in particular, are high in phenolics. During applesauce and canned apple manufacture, the antioxidant-rich peels of apples are discarded. To determine if a useful source of antioxidants is being wasted, the phytochemical content, antioxidant activity, and antiproliferative activity of the peels of four varieties of apples (Rome Beauty, Idared, Cortland, and Golden Delicious) commonly used in applesauce production in New York state were investigated. The values of the peels were compared to those of the flesh and flesh + peel components of the apples. Within each variety, the total phenolic and flavonoid contents were highest in the peels, followed by the flesh + peel and the flesh. Idared and Rome Beauty apple peels had the highest total phenolic contents (588.9 +/- 83.2 and 500.2 +/- 13.7 mg of gallic acid equivalents/100 g of peels, respectively). Rome Beauty and Idared peels were also highest in flavonoids (306.1 +/- 6.7 and 303.2 +/- 41.5 mg of catechin equivalents/100 g of peels, respectively). Of the four varieties, Idared apple peels had the most anthocyanins, with 26.8 +/- 6.5 mg of cyanidin 3-glucoside equivalents/100 g of peels. The peels all had significantly higher total antioxidant activities than the flesh + peel and flesh of the apple varieties examined. Idared peels had the greatest antioxidant activity (312.2 +/- 9.8 micromol of vitamin C equivalents/g of peels). Apple peels were also shown to more effectively inhibit the growth of HepG(2) human liver cancer cells than the other apple components. Rome Beauty apple peels showed the most bioactivity, inhibiting cell proliferation by 50% at the low concentration of 12.4 +/- 0.4 mg of peels/mL. The high content of phenolic compounds, antioxidant activity, and antiproliferative activity of apple peels indicate that they may impart health benefits when consumed and should be regarded as a valuable source of antioxidants.
Differences in survival and growth among five different Escherichia coli O157:H7 strains in three apple varieties were determined at various temperatures. Jonathan, Golden Delicious, and Red Delicious apples were wounded and inoculated with E coli O157:H7 strains C7929 (apple cider isolate), 301C (chicken isolate), 204P (pork isolate), 933 (beef isolate), and 43890 (human isolate) at an initial level of 6 to 7 log CFU/g. The inoculated apples were stored at a constant temperature of 37, 25, 8, or 4 degrees C or at 37 degrees C for 24 h and then at 4 degrees C, and bacterial counts were determined every week for 28 days. By day 28, for Jonathan apples at 25 degrees C, the apple isolate counts were significantly higher than the chicken and human isolate counts. At 4 degrees C for 28 days, the human isolate inoculated into Jonathan, Golden Delicious, and Red Delicious apples was present in significantly smaller numbers than the other strains. The apple isolate survived significantly better at 4 degrees C, yielding the highest number of viable cells. By days 21 and 28, for apples stored at 37 degrees C for the first 24 h and then at 4 degrees C, the counts of viable E. coli O157:H7 apple and human isolates were 6.8 and 5.8 log CFU/g at the site of the wound, whereas for apples kept at 4 degrees C for the duration of storage, the respective counts were 5.6 and 1.5 log CFU/g. Our study shows that E. coli O157:H7 strains responded differentially to their ability to survive in these three apple varieties at 25 or 4 degrees C and produced higher viable counts when apples were temperature abused at 37 degrees C for 24 h and then stored at 4 degrees C for 27 days.
A Drosophila gene [amyloid protein precursor-like (Appl)] has recently been identified whose predicted amino acid sequence (APPL) shares extensive homology with the beta-amyloid protein precursor (APP) associated with Alzheimer's disease. Characterization of proteins encoded by the Appl gene was initiated with the expectation that this simple model system might help elucidate the basic function provided by APPL and APP proteins. In this report, we identify 2 forms of the APPL protein in embryonic extracts, primary cultures, and transfected cells. APPL is synthesized as a 145-kDa membrane-associated precursor that is converted to a 130-kDa secreted form that lacks the cytoplasmic domain. Both forms are N-glycosylated. Pulse-chase and subcellular localization studies suggest that the conversion is very rapid. The similarities of biogenesis between APP and APPL provide further evidence that APPL and APP might be functionally homologous, and that the secretion event is of physiological significance. Immunocytochemical studies show that the APPL proteins are first detected in developing neurons concomitant with axonogenesis and remain associated with differentiated neurons. APPL immunoreactivity was observed in neuronal cell bodies, axonal tracts, and neuropil regions. In the embryo, APPL proteins are expressed exclusively in the CNS and PNS neurons, consistent with the Appl transcript localization. The expression pattern of APPL proteins suggests an ancestral function for this protein in the nervous system.
Focusing on 17 constituents, the polyphenol profiles of juices freshly made from various dessert (n = 4) and cider apple cultivars (n = 7) as well as commercially available apple juices (n = 24) were investigated using high-performance liquid chromatography-photodiode array detection (HPLC-DAD) and (HPLC)-electrospray ionization-tandem mass spectrometry (ESI(neg)-MS/MS) analyses. Significant differences in the total polyphenol content as well as the profiles of the apple cultivars under study were observed. For dessert apples the total polyphenol content ranged from 154 to 178 mg/L, whereas for 'old' German cider apple cultivars 261-970 mg/L were determined. Boskoop showed the highest (970 mg/L) and Granny Smith the lowest (154 mg/L) polyphenol content of the freshly prepared samples under study. Hydroxycinnamic acids, with chlorogenic acid as dominating constituent, ranged from 57 to 68 mg/L as well as from 134-593 mg/L in juices made from dessert apples and that from cider apples, respectively. Dessert apple juices showed lower contents of dihydrochalcones (10-35 mg/L) and flavan-3-ols (50-95 mg/L) compared to that of cider apples (34-171 mg/L and 70-393 mg/L, respectively). Quercetin and its derivatives were found from 0.4-4 mg/L and 0.4-27 mg/L in juices made from dessert apples and that of cider apples, respectively. Compared with freshly made juices, lower contents of polyphenols were determined in the commercial samples under study. Amounts ranging from 110-459 mg/L, dominated by chlorogenic acid with concentrations from 53-217 mg/L, were determined. Information about cultivar-typical apple polyphenol content and profile is important for bioactivity studies and, consequently, essential for the development of consumer-relevant products with particular nutritional functionalities.
Survival of Escherichia coli O157:H7, Salmonella Muenchen, and yeasts and molds on apples as affected by application of five commercial apple waxes was investigated. Red Delicious cv. apples at 21 degrees C were spot inoculated with E. coli O157:H7 and S. Muenchen and spray coated with waxes. Apples sprayed with water served as controls. Apples were dried at either 21 or 55 degrees C for 2 min before subjecting to microbiological analysis after storage for 0, 1, 3, 6, and 12 weeks at 2 or 21 degrees C. Drying temperature did not significantly influence populations of E. coli O157:H7 and S. Muenchen. Waxing reduced populations E. coli O157:H7 and S. Muenchen by up to 1.48 log10 cfu/apple. Compared to untreated apples, treatment of apples with water or waxes resulted in significant (P < or = 0.05) reductions in populations of E. coli O157:H7 and S. Muenchen during storage at 2 degrees C. Reductions on waxed apples stored at 21 degrees C were not as marked compared to reductions on waxed apples stored at 2 degrees C. With the exception of one wax, drying temperature did not significantly influence populations of yeasts and molds. Mold populations were less affected by wax applications than were yeasts, and were detected in higher numbers on apples treated with three of the five waxes compared to populations recovered from untreated control apples. None of the waxes evaluated can be relied upon to kill or remove E. coli O157:H7 and Salmonella on apples.
BACKGROUND: Most patients with birch pollen allergy report oral allergy symptoms after eating fresh apples and other vegetable foods. Major birch pollen and apple allergens, Bet v 1 and Mal d 1, are highly homologous; as a consequence, pollen-specific immunotherapy (SIT) might be expected to improve apple hypersensitivity. OBJECTIVE: To evaluate the clinical and immunological effects of birch pollen SIT on oral allergy syndrome (OAS) induced by apples. METHODS: A prospective study carried out in 49 birch pollen-sensitive patients with apple-induced OAS who received injection immunotherapy for 12, 24, or 36 months. Twenty-six patients not submitted to SIT and followed up for 12-48 months were used as controls. Both SPT and open oral challenges with fresh golden delicious apple were performed, as well as specific IgE measurements, before and after SIT. RESULTS: Forty-one patients (84%) vs no control (0%) reported a significant reduction (50-95%) or a total disappearance (100%) of OAS symptoms after SIT (P < 0. 001). Similar responses were observed in patients treated for 12, 24, or 36 months. SIT also induced a marked reduction in skin reactivity against fresh apple in 43 patients (88%). The effect of SIT was inversely related with baseline skin reactivity: 50% and 8% patients with a weakly or strongly positive baseline apple skin prick tests (SPT), respectively, did not report changes in OAS severity after SIT (P < 0.01). In contrast, baseline birch pollen-specific or apple-specific IgE antibodies levels did not influence SIT effectiveness on OAS. SIT induced a marked decrease in birch pollen-specific IgE levels (P < 0.001), whereas apple-specific IgE showed an unexpected variability (reduction in 21%, no change in 43%, increase in 38%). No control subject reported a reduction in OAS severity or showed a decrease in skin reactivity at follow-up (P < 0.001). CONCLUSIONS: SIT with birch pollen extracts effectively reduces clinical apple sensitivity and skin reactivity in most cases after only 1 year of treatment; these effects are not paralleled by a similar reduction in apple-specific IgE. These findings suggest a decrease in activability of effector cells as the mechanism underlying clinical benefit.
Marinades containing 0, 0.4, 0.8, and 1.2% apple flavoring were examined to determine the effect of an aqueous apple flavoring on the quality and sensory characteristics of boneless, skinless chicken breast. Marinade pickup and purge loss increased significantly with an increase of apple flavoring in marinades. Incorporation of apple flavoring in the marinades did not affect cook loss of the treatments, except for breast marinated in 1.2% apple flavor, which had higher cook loss. Apple flavoring did not affect final product yield of marinated breast. Shear values were similar for all treatments except marinades with 0.8% apple flavoring, which had lower shear values. Participants detected increases in fruity flavor with an increase of apple flavoring in the marinade. Products containing no apple flavoring were rated as "like moderately" to "like slightly." Products with 0.4% apple flavoring were rated "like slightly," and degree of liking declined to "neither like nor dislike" as apple flavoring increased. Incorporation of acceptable levels of apple flavoring is limited to 0.4%.
Four of five apple cultivars (Golden Delicious, Red Delicious, McIntosh, Macoun, and Melrose) inoculated with Escherichia coli O157:H7 promoted growth of the bacterium in bruised tissue independent of the date of harvest (i.e., degree of apple ripening) or the source of the apple (i.e., tree-picked or dropped fruit). Apple harvest for this study began 4 September 1998 and ended 9 October, with weekly sampling. Throughout this study, freshly picked (<2 days after harvest) McIntosh apples usually prevented the growth of E. coli O157:H7 for 2 days. Growth of E. coli O157:H7 did occur following 6 days of incubation in bruised McIntosh apple tissue. However, the maximum total cell number was approximately 80-fold less than the maximum total cell number recovered from Red Delicious apples. When fruit was stored for 1 month at 4 degrees C prior to inoculation with E. coli O157:H7, all five cultivars supported growth of the bacterium. For each apple cultivar, the pH of bruised tissue was significantly higher and degrees Brix was significantly lower than the pH and degrees Brix of undamaged tissue regardless of the source. In freshly picked apples, changes in the pH did not occur over the harvest season. Bruised Golden Delicious, McIntosh, and Melrose apple tissue pHs were not significantly different (tree-picked or dropped), and the degrees Brix values of McIntosh, Macoun, and Melrose apple tissue were not significantly different. Single-cultivar preparations of cider did not support growth of E. coli, and the cell concentration of inoculated cider declined over an 11-day test period. The rate of decline in E. coli cell concentration in the McIntosh cider was greater than those in the other ciders tested. The findings of this study suggested that the presence of some factor besides, or in addition to, pH inhibited E. coli growth in McIntosh apples.
Confocal scanning laser microscopy (CSLM) was used to determine the location of Escherichia coli O157:H7 cells on the surface and in tissue of bruised Red Delicious cv. apples. Undamaged and bruised apples were inoculated by immersing in a suspension of E. coli O157:H7 cells transformed with a plasmid that encodes for the production of a green fluorescent protein. Apples were then washed in 0.1% (wt/vol) peptone water and/or rubbed with a polyester cloth and examined to determine if these treatments removed or introduced cells into lenticels, cutin, and cracks on the skin surface. Optical slices of the apples obtained using CSLM were examined to determine the depth at which colonization or attachment of cells occurred. Populations of E. coli O157:H7 on the surface of apples were determined to assess the effectiveness of washing and rubbing in physically removing cells. The location of cells on or in undamaged and bruised areas of apples that were not washed or rubbed did not differ significantly. However, washing apples resulted in an approximate 2-log reduction in CFU of E. coli O157:H7 per cm2 of apple surface. On unwashed apples, cells were detected at depths up to 30 microm below the surface. No E. coli O157:H7 cells were detected at locations more than 6 microm below the surface of washed apples. Cells that remained on the surface of rubbed apples appeared to be sealed within naturally occurring cracks and crevices in waxy cutin platelets. These cells may be protected from disinfection and subsequently released when apples are eaten or pressed for cider production.
BACKGROUND: Due to the cross-reactivity between Bet v 1 and proteins present in vegetable foods, birch pollen allergic patients frequently develop allergy to fruits and vegetables, mostly apples. Since many apple cultivars exist some of them might contain sufficiently low amounts of Mal d 1 to be tolerated by most allergic patients. OBJECTIVE: To assess whether apple cultivars containing low amounts of Mal d 1 are better tolerated by apple-allergic patients. METHODS: Mal d 1 content was determined in many apple cultivars by ELISA. Selected cultivars containing high (Golden Delicious) or low (Orim, G 198 and Vienna) amounts of Mal d 1 were compared in apple allergic patients both by SPT and oral challenges. RESULTS: The 3 different apple cultivars induced wheals of similar size in most patients. Upon oral challenges no patient reported the total absence of oral symptoms following the ingestion of either high or low allergenic apples. Golden Delicious and G-198 elicited OAS of similar severity 3/7 cases. The 2 cultivars induced significantly more severe symptoms in 2 cases each. CONCLUSION: Allergy to Mal d 1 is characterized by significant inter-patient variability. Moreover, marked inter-apple and intra-apple variability exists. As a consequence, the amount of Mal d 1 in apples classified as containing low concentrations of allergen may be sufficient to induce both clinical symptoms and skin reactivity in birch pollen-allergic patients. The search for low allergenic apples therefore should be continued and extended to other germplasm accessions, be it cultivars, breeding lines or wild species.