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Long-term hypolipaemic effect of apple pulp on golden hamsters with alimentary hyperlipoproteinaemia.

Golden hamsters with alimentary hyperlipoproteinaemia (fed on a diet in which 44% of the energy was accounted for by fats, 52 mg cholesterol (CH)/100 g) were given apple pulp (10% of the diet) for six months. From the end of the first month up to the end of the experiment, elevation of serum CH, triacylglycerol (TG) and phospholipid (PL) levels and CH and TG accumulation in the liver were effectively inhibited. At the close of the experiment, hyperlipoproteinaemia was characterized by an increase in the amount of very low density lipoproteins (VLDL), which carried 60-80% of all circulating lipids, accounted for over 70% of the total lipoprotein pool and transported almost 60% of the serum CH (high density lipoproteins--HDL--only 16%). The apple diet markedly reduced (by 50-80%) the amount of all lipids in the lipoproteins within density limits of 1,006-1,063 kg/m3 and lowered the total lipid concentration in these density classes and in the lipoprotein pool as a whole (by 50%). The proportion of the individual lipoprotein classes in the lipoprotein pool was not significantly affected. The apple diet did not affect either the chemical composition of HDL or their participation in the lipoprotein pool, but raised the proportion of serum CH transported in this lipoprotein class. Neither the lipoprotein lipase activity of the adipose tissue nor lecithin:cholesterol acyltransferase activity were influenced by the apple diet.

Adipose Tissue↗

[Isolation and partial characterization of phenoloxidase from apples (Malus domestica, var. Anna)].

This study pursued the isolation and partial characterization of the enzyme polyphenoloxidase from apple (Malus domestica Anna variety), grown in the Hermosillo Coast (State of Sonora, Mexico). The effects of pH and temperature as well as its specificity towards substrates, and its behavior under conditions of hydrophobic chromatography, were studied. The enzyme was isolated from a residual powder obtained from ripe apples homogenized with cold acetone. The extract thus prepared was used to characterize the enzyme, and it showed an optimum pH of 5.36 and an optimum temperature of 35 degrees C. The substrate specificity proved to decrease from 4-methyl catechol, chlorogenic acid, catechol, and caffeic acid, to 3,4-dihydroxiphenyl alanine (DOPA). The enzyme resulted to be more thermostable (temperature range: 35 degrees C to 60 degrees C) than the rest of oxidases of plant origin. When the extract was eluted under conditions of hydrophobic chromatography separation, it appeared as a single peak resulting in a 300 fold purification. The phenolase activity characteristics found in the present study were similar to those observed in other apples from temperate climates; however, this particular polyphenoloxidase is more thermostable under natural conditions. This explains why apples of the Anna variety, at the high harvesting temperature, show a very fast formation of brown spots even when there is a minor damage. The content of compounds with phenolic group was high (1.16 g/100 g fresh weight). Further increase of the velocity of fruit enzymatic browning was due to this reason.

Chromatography↗

Stable isotopic carbon composition of apples and their subfractions--juice, seeds, sugars, and nonvolatile acids.

The 13C:12C ratios of 8 authentic apple juice samples and their subfractions were determined by mass spectrometry. Apples from Argentina, Mexico, New Zealand, and the United States were processed into juice; pulp was collected from the milled fruit and seeds were collected from the press-cake. Sugars, nonvolatile acids, and phenolics were isolated from the juice by treatment with ion-exchange resins and polyvinylpyrrolidone (PVPP). The mean value for all juice samples was -24.2% which is close to the values reported by other investigators. Juice from apples grown in Argentina, Mexico, and New Zealand did not differ from U.S. samples. The isotopic composition of the subfractions ranged from -22.0 to -31.0%. The values for the pulp were essentially the same as for juice. The sugar fraction was slightly less negative than the juice; the nonvolatile acid and phenolic fractions were more negative. The levels of nonvolatile acids and phenolics in apple juice are low, however, so these compounds contribute little to overall delta 13C values in juice.

Acids↗

[Investigations about cause of specific replant disease of fruit trees. VI. Proof of actinomycetes in feeder roots of apple seedlings in soils with different digrees of soil sickness (author's transl)].

Microscopic analysis of longitudinal sections of feeder roots of apple seedlings from soil with experimentally prepared different degrees of soil sickness led to the following results: 1. A colonization of root-barks by actinomycetes could be pointed out in injured and in macroscopic not visible injured feeder roots of apple seedlings from sick soil. 2. The frequency of the colonization of root-pieces of the two highest root classes was in the sick soil at 47.3% and in the two soils with a less degree of soil sickness at 32.4%. In the non-sick soil the root were at a frequency of 0.3% nearly free from actinomycetes. 3. Feeder roots colonizated by actinomycetes showed injuries in the cell-bandage. Increasing injuries of the roots were obtained with increasing intensity of colonization by actinomycetes. In heavy injured feeder roots the proof of actinomycetes was difficult or no more possible. As the share of heavy injured roots was increasing with increasing degree of soil sickness can be supposed that the real frequency of root colonization by actinomycetes is substantially higher than it was observed in roots seeming to be healthy macroscopically. 4. The results obtained support our hypothesis that soil sickness of apple trees is caused by actinomycetes. The injuries of the roots appearing as a consequence of the colonization may explain both, the soil sickness in the case of replanting and the development of soil sickness in growing apple plantations. 5. Since until now there is no proof that actinomycetes cause injuries in roots of fruit-trees further examinations have to be aspired to confirm our results.

Actinomycetales↗

[Apple powder in the treatment of patients with chronic enteritis].

A manifest clinical response has been achieved in 38 patients with chronic enteritis kept on apple diet in the form of apple powder produced from apple juice refuse. Carbohydrate loading with starch (polysaccharide), saccharose (disaccharide), glucose (monosaccharide), d-xylose made it clear that the apple powder improves hydrolysis and carbohydrate absorption: by 30%, 23%, 32% and 40% for starch, saccharose, glucose and d-xylose, respectively. Attenuation of the inflammation in the small intestine was also evident from the tendency to normalization of some fecal intestinal enzymes activity (entero-kinase, alkaline phosphatase). Changes in the systems PGE-cAMP and PGF-cGMP are suggested to play a role in the emergence of malabsorption syndrome, diarrhea, structural lesions in small intestinal mucosa.

Alkaline Phosphatase↗

Detection of adulteration in apple juice by L-malic/total malic acid ratio: collaborative study.

L-Malic acid is the predominate acid in pure apple juice and no D-malic acid should be present. Synthetic malic acid contains 50% D-malic acid, is inexpensive, and can be used to create nonauthentic apple juice. L-Malic/total malic ratios of 0.9 or less are indicative of a nonauthentic sample. Fourteen laboratories participated in a collaborative study to determine the L-malic/total malic acid ratio in apple juice. Ten samples of apple juice were sent to each laboratory. Authenticity of the samples varied from 0 to 100%. The coefficients of variation in all cases were acceptable, i.e., ca 5%. The method was adopted first action by AOAC INTERNATIONAL.

Beverages↗

[The effect of preoperative oral fluid intake on the volume and pH of gastric contents in elective surgical patients--a comparison of tea with apple juice].

This study aimed to investigate the effect of 150 ml of either tea or apple juice on the volume and pH of gastric contents in 40 elective surgical patients, ranging in age from 18 to 70 years. They were given diazepam 5 approximately 10 mg and roxatidine 75 mg orally 2 hours before the start of isoflurane anesthesia or modified neuroleptic anesthesia. Immediately following the induction of anesthesia with thiopental and vecuronium, a nasogastric tube was placed to aspirate the gastric content to measure its volume and pH. The volume and pH of gastric contents were 6.4 +/- 8.5 ml, 6.0 +/- 2.2 in tea group and 17.1 +/- 18.4 ml, 4.4 +/- 2.6 in apple juice group, respectively. There was a significant difference in the gastric volume between the two groups (P < 0.05), while no significant difference in gastric pH was observed. This result suggests that apple juice is not appropriate as preoperative drink because apple juice increases gastric contents, and may cause aspiration pneumonia.

Adolescent↗

Determination of patulin in apple juice by liquid chromatography: collaborative study.

An AOAC International-International Union of Pure and Applied Chemistry-International Fruit Juice Union (AOAC-IUPAC-IFJU) collaborative study was conducted to evaluate a liquid chromatographic (LC) procedure for determination of patulin in apple juice. Patulin is a mold metabolite found naturally in rotting apples. Patulin is extracted with ethyl acetate, treated with sodium carbonate solution, and determined by reversed-phase LC with UV detection at 254 or 276 nm. Water, water-tetrahydrofuran, or water-acetonitrile was used as mobile phase. Levels determined in spiked test samples were 20, 50, 100, and 200 micrograms/L. A test sample naturally contaminated at 31 micrograms/L was also included. Twenty-two collaborators in 10 countries analyzed 12 test samples of apple juice. Recoveries averaged 96%, with a range of 91-108%. Repeatability relative standard deviations (RSDr) ranged from 10.9 to 53.8%. The reproducibility relative standard deviation (RSDR) ranged from 15.1 to 68.8%. The LC method for determination of patulin in apple juice has been adopted first action by AOAC INTERNATIONAL.

Beverages↗

Evaluation of recombinant allergens Bet v 1 and Bet v 2 (profilin) by Pharmacia CAP system in patients with pollen-related allergy to birch and apple.

Sixty-five patients presenting either rhinoconjunctivitis or asthma and sensitized to pollens of trees of the order Fagales were studied by the Pharmacia CAP system in order to assess specific IgE for the important birch pollen allergens Bet v 1 and Bet v 2. All 65 subjects reacted to at least one of the recombinant birch allergens: 43% to Bet v 1, 30.7% to Bet v 2, and 26% to both. Patients monosensitized to birch did not react to Bet v 2. Of patients with a history of oral allergy syndrome after eating apples, 16/28 (57%) reacted to Bet v 1; among 20 polysensitized subjects presenting oral allergy syndrome after consumption of apple, four reacted to Bet v 2 (20%). Among patients with IgE against both recombinant allergens, six (35.30%) presented symptoms of allergy after eating apples. Our results indicate that sensitization to Bet v 1 is specific for birch and apple allergies, whereas sensitization to Bet v 2 is common in polysensitized patients.

Adolescent↗

[Repetitive movements of the upper limbs: results of exposure evaluation and clinical investigation during the sorting and packaging of apples].

In the fruit industry, workers sorting and packing apples perform a job that is characterised by repetitive actions, that require them to remain in an upright position for prolonged periods of time and, occasionally, to manually lift loads of apples. The aim of the study was to detect and provide a preliminary quantification of any possible risks for the musculo-skeletal system, as well as any disorders present among the workers. A risk analysis for WMSDs was performed in an apple packing plant, using video recordings of the job cycles associated with two different tasks (semi-automatic sorting and manual packing). A clinical and anamnestic investigation was carried out in nine plants; a standard questionnaire was distributed to 180 employees, with the purpose of detecting disorders and diseases of the locomotor system. An initial evaluation of the risks for WMSDs indicated that the routine tasks performed by the female apple workers featured a very high frequency of actions, also in view of the fact that the repetitive tasks lasted for the duration of the entire work shift. In the situation examined, the action frequency limits were greatly exceeded; moreover, the recovery times were inadequate in terms both of their duration and distribution. Regarding clinical and anamnestic examination, an overall analysis of the results shows a very high prevalence of spinal and hand-arm disorders. Given these preliminary findings, the authors suggest that the sample population of fruit sorters should undergo clinical and instrumental testing to evaluate the condition of the spine and upper limbs, and that a specific health surveillance programme should be adopted across the industry.

Adult↗

MdWRKY75 interacts with MdWOX11 to modulate root growth under salt stress in apple.

The root system is pivotal for plant development, enabling both vegetative growth and tolerance to abiotic stresses like salinity. However, the molecular mechanisms governing root adaptive development in response to salt stress remain poorly understood in apple (Malus domestica Borkh.). In this study, we identified the salt stress-responsive WRKY transcription factor MdWRKY75. Overexpression of MdWRKY75 in transgenic apple negatively regulates adventitious root (AR) formation and salt stress tolerance, whereas reducing MdWRKY75 expression yields the opposite phenotype. Moreover, MdWRKY75 directly binds to the promoter of MdSAUR15 (SMALL AUXIN UP RNA15) and transcriptionally represses the expression of MdSAUR15, which, when overexpressed, promotes AR formation and enhances salt stress tolerance. We further demonstrated that MdWRKY75 interacts with MdWOX11, a WUSCHEL-related homeobox (WOX) transcription factor, both in vitro and in vivo. MdWOX11 expression is upregulated and enhances AR formation under salt stress. Additionally, MdWOX11 reduces the binding of MdWRKY75 to the MdSAUR15 promoter, and alleviates the MdWRKY75-mediated inhibitory effect on MdSAUR15 expression. Collectively, our study provides a MdWOX11-MdWRKY75-MdSAUR15 module regulating root adaptation in response to salt stress in apple.

Malus↗

The effect of dried tomato, grape and apple pomace on the cholesterol metabolism and antioxidative enzymatic system in rats with hypercholesterolemia.

The effect of dried tomato, grape and apple pomace (5% in diet) on the cholesterol levels in serum and selected organs was studied in male Wistar rats fed a cholesterol diet (0.3%) shortly after weaning. Pomace of individual fruits contained 47.2, 67.0 and 46.2 g of total dietary fiber per 100 g of dry matter for tomato, grape and apple pomace, respectively. Corresponding values for the soluble components of the fiber were 10.3, 14.7 and 26.0%. None of the tested products affected the serum cholesterol levels. Tomato and apple pomace significantly reduced cholesterol level in liver by 15% and 11%, respectively, and all tested pomace species reduced cholesterol content in heart (by 18-21%). Triacylglycerol content was affected only by tomato pomace and solely in the heart. Cholesterol absorption measured by dual-isotope ratio method, was not significantly affected by the pomace, however, all products strongly reduced (up to the half of the original value) the activity of HMG-CoA reductase in liver and increased the fractional catabolic rate of plasma cholesterol (by 60-120%). All tested pomace reduced plasma levels of conjugated dienes and tomato and grape pomace exhibited this effect also in the liver. All tested pomace reduced the activities of antioxidant enzymes superoxid dismutase, catalase and glutathione peroxidase in erythrocytes (by 34-56%).

Animal Feed↗

Characteristics and physiological functions of polyphenols from apples.

Apples contain many kinds of polyphenols, and the main components are oligomeric procyanidins. Applephenon is apple polyphenol extract produced commercially from unripe apples, and has been used as food additive in order to prevent oxidation of components in foods and its application in functional foods is expected. In a lipid metabolism regulation study, administration of Applephenon has the potential to exert strong anti-oxidative activity and to inhibit consumption of vitamin E and anti-oxidative enzymes. Double blind clinical trials of Applephenon on pediatric patients with atopic dermatitis, and tests using type I allergic model mice suggested that Applephenon might regulate allergic reactions. We found the no observed adverse effect level (NOAEL) of Applephenon is greater than 2000 mg/kg in a 90~day consecutive oral administration toxicity test in rats, and Applephenon is safe and acceptable based on mutagenicity tests.

Animals↗

[Retention of apple starch in production of pectin by the aluminum pectinate procedure].

Apple pectin produced by means of the aluminum pectinate procedure contains less starch than apple pectin obtained by precipitation with ethanol. The degradatin of the starch which occurs during production depends to a great extent upon the conditions of pomace digestion. The absorption maxima of the amylose-iodine complexes range in general from 550 to 570 nm. Model experiments with highly esterified pectin and partially degraded (by mechanolysis) apple starches were performed to establish favourable conditions for fractionating the pectin and starch components which are obtained when the pectin is precipitated as aluminum pectinate and when the granular coagulate is washed with water.

Aluminum↗

Determination of norethindrone stability on red delicious apples.

A stability-indicating assay procedure for the determination of norethindrone on the surface of Red Delicious apple slices was developed. The apple slice is homogenized in pH 10 borate buffer solution and norethindrone is then partitioned into chloroform using a diffusion chamber that has a membrane that is permeable to hydrophobic compounds separating the buffer and chloroform. An aliquot of the chloroform extract is then evaporated to dryness and reconstituted in tetrahydrofuran. The reconstituted sample is then chromatographed on a C18 reversed-phase column using a mobile phase consisting of water:tetrahydrofuran:methanol (63:26:11, v/v/v). Detection is in the UV range at 254 nm. The chromatographic system is specific for norethindrone in the presence of its autoxidation products. Stability data indicate that norethindrone is stable for up to 6 h on the surface of Red Delicious apples.

Chromatography, High Pressure Liquid↗

Modulation of oxidative cell damage by reconstituted mixtures of phenolic apple juice extracts in human colon cell lines.

Diets rich in fruits and vegetables are associated with a lower risk of tumour induction in the intestine and other sites. Apple juice with high amounts of antioxidative phenolics might protect the intestine against reactive oxygen species-mediated cell damage. We investigated to which extent the preventive effectiveness of polyphenolic juice extracts is governed by the amounts of five major constituents (rutin, phloridzin, chlorogenic acid, caffeic acid and epicatechin). In human colon cell lines (Caco-2, HT29), reconstituted mixtures of these phenolics were investigated in comparison to the original juice extracts, originating from cider and table apples. Parameters studied were (oxidative) DNA damage (Comet assay), cellular redox status (dichlorofluorescein assay) and Trolox equivalent antioxidant capacity (TEAC). The TEAC of the reconstituted mixtures was higher compared to the respective original extracts (4.7-7.3 mM vs. 3.6-4.2 mM Trolox). After 24 h cell incubation, menadione-induced (oxidative) DNA damage was more effectively reduced by the reconstituted mixtures (1-100 microg/mL, 24 h), as compared to the original extracts. In contrast, the cellular ROS level was reduced to a rather similar extent by original extracts and reconstituted mixtures. The results lead to the conclusion that the selected constituents in their authentic proportions substantially account for the antioxidative effectiveness of phenolic apple juice extracts.

Antioxidants↗

Studies on apple and blueberry fruit constituents: do the polyphenols reach the colon after ingestion?

The aim of our studies was to determine the amount of polyphenols reaching the colon after oral intake of apple juice and blueberries. After a polyphenol-free diet healthy ileostomy volunteers consumed a polyphenol-rich cloudy apple juice while others consumed anthocyanin-rich blueberries. Ileostomy effluent was collected and polyphenols were identified using HPLC-DAD as well as HPLC-ESI-MS/MS; quantification was performed with HPLC-DAD. Most of the orally administered apple polyphenols were absorbed from or metabolized in the small intestine. Between 0 and 33% of the oral dose was recovered in the ileostomy bags with a maximum of excretion after 2 h. A higher amount of the blueberry anthocyanins under study (up to 85%, depending on the sugar moiety) were determined in the ileostomy bags and therefore would reach the colon under physiological circumstances. Such structure-related availability has to be considered when polyphenols are used in model systems to study potential preventive effects in colorectal diseases.

Anthocyanins↗

Detection of isorhamnetin glycosides in extracts of apples (Malus domestica cv. "Brettacher") by HPLC-PDA and HPLC-APCI-MS/MS.

Extracts of apple fruits (Malus domestica cv. "Brettacher") were analysed by HPLC with photodiode array detection. An unknown peak was monitored displaying the same retention time as isorhamnetin 3-O-glucoside. Preliminary identification of the isorhamnetin aglycone was performed by comparison of UV spectral data of the unknown compound with a reference substance. Using atmospheric pressure chemical ionisation mass spectrometry in the negative ion mode, the presence of an isorhamnetin glycoside was supported by loss of 162 amu from the pseudomolecular ion (m/z 477). MS2 product ion analysis of the parent ion m/z 477 provided a fragmentation pattern identical to the reference. Collision-induced dissociation of the aglycone (m/z 315) in the MS3 product ion analysis allowed the differentiation of rhamnetin and isorhamnetin, and unambiguous assignment by comparison with standard compounds. A second isorhamnetin glycoside eluting prior to the glucoside was tentatively identified as isorhamnetin 3-O-galactoside. To the best of our knowledge, this is the first report of isorhamnetin glycosides in apple fruit extracts. Results are discussed with respect to chemotaxonomic relevance within the genera Malus and Pyrus, and especially in consideration of the control of the authenticity of apple products.

Atmospheric Pressure↗