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PubMed2004

The toxicology and safety of apple polyphenol extract.

Apple polyphenol extract has strong antioxidant activity and various physiological functions, and is used in Japan as a food additive and nutritional supplements. Here, we tested the consumption safety of Applephenon, which is a polyphenol extract produced from unripe apples. The Ames test without S9 mixture revealed that Applephenon, had slight mutagenicity at a high concentration of 2500 microg/plate; however, both chromosomal aberration test and the micronucleus test found no significant mutagenicity. Furthermore, an acute oral-toxicity test, and a 90-day subchronic-toxicity test showed no significant hematological, clinical, chemical, histopathological, or urinary effects at a dose of 2000 mg/kg. These results confirm that Applephenon is safe and no toxic at average dietary level.

Administration, Oral↗
PubMed2005

Oxoester oxidoreductase activities in new isolates of Pichia anomala from apple, grape and cane juices.

Thirty-nine yeasts isolated from apple, grape and cane juices were screened for their oxidoreductase activity. The two strains of Pichia, one isolated from apple and one from cane juices, appear to be promising strains for oxidoreductase activity on alpha-oxoesters. They showed similar high yields in converting ethyl pyruvate to ethyl lactate as Saccharomyces spp. (86.6% and 85.3% versus 86.6%), and higher yields in the reduction of alpha-oxocarboxylic esters (ketopantolactone to pantolactone: 74% and 73.3%, respectively) compared to Saccharomyces spp. (yield 60%).

Beverages↗
PubMed2004

Simultaneous silencing of multiple genes in the apple scab fungus, Venturia inaequalis, by expression of RNA with chimeric inverted repeats.

RNA-mediated gene silencing has been demonstrated in plants, animals, and more recently in filamentous fungi. Here, we report high frequency, RNA-mediated gene silencing in the apple scab fungus, Venturia inaequalis. The green fluorescent protein (GFP) transgene was silenced in a GFP-expressing transformant. An endogenous gene, trihydroxynaphthalene reductase (THN), involved in melanin biosynthesis, was also silenced. Silencing of these two genes resulted in obvious phenotypes in vitro. High frequency gene silencing was achieved using hairpin constructs for the GFP or the THN genes transferred by Agrobacterium (71 and 61%, respectively). THN-silenced transformants exhibited a distinctive light brown phenotype and maintained the ability to infect apple. Of significance was the simultaneous silencing of the two genes from a single chimeric, inverted repeat hairpin construct. Silencing of both genes with this construct occurred at a frequency of 51% of all the transformants. All 125 colonies silenced for the GFP gene were also silenced for THN. As THN and GFP silenced transformants have readily detectable phenotypes, the genes have utility as markers for gene silencing. Simultaneous, multiple gene silencing, utilising such marker genes, will enable the development of high through-put screening for functional genomics. This chimeric technology will be particularly valuable when linked with silenced genes that have no obvious phenotype in vitro.

Ascomycota↗
PubMed2005

Two types of spermine synthase gene: MdACL5 and MdSPMS are differentially involved in apple fruit development and cell growth.

Three cDNAs with high homology to spermine (Spm) synthases in Arabidopsis were isolated from apple [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.]. MdACL5-1 and MdACL5-2 have high homology with ACL5 and MdSPMS has high homology with AtSPMS. The similarity of MdSPMS to spermidine synthases (SPDSs) was higher than that of MdACL5s, despite the fact that both are putative Spm synthases. However, MdSPMS could be discriminated from SPDSs by the presence of several characteristic amino acids, i.e., Val-149, Ser-161, Ala-205, and Val-235, in the decarboxylated S-adenosylmethionine (dcSAM)-binding motif of MdSPMS. Both MdACL5-1 and MdSPMS complemented Spm biosynthesis in a yeast mutant deficient in Spm synthase, and ectopic expression of MdACL5-1 in the Arabidopsis dwarf mutant acl5 allowed recovery of the normal phenotype. RNA gel blot analysis showed that MdACL5 and MdSPMS are differentially expressed in tissues and suspension cells. These results suggest that functional MdACL5 and MdSPMS are independently involved in apple fruit development and cell growth.

Amino Acid Sequence↗
PubMed2005

Rapid discrimination of Alicyclobacillus strains in apple juice by Fourier transform infrared spectroscopy.

Alicyclobacillus spp. are thermoacidophilic, spore-forming bacteria. Some of which cause spoilage in pasteurized and heat-treated apple juice products through the production of guaiacol. Fourier transform infrared (FT-IR) spectroscopy was used to discriminate between eight Alicyclobacillus strains (WAC, 81-2, Oly#21, 51-1, KF, 1016, 1101, and A-Gala A4) in apple juice. FT-IR vibrational combination bands reflected compositional differences in the cell membranes of Alicyclobacillus strains in the "fingerprint region" at wavenumbers between 1500 and 800 cm(-1). Distinctive segregation among spectral sample clusters of different Alicyclobacillus strains was observed using principal component analysis (PCA). Two closely related strains (1016 and 1101) of Alicyclobacillus acidoterrestris could be distinguished, suggesting that this method can be highly selective. Results of soft independent modeling of class analogy (SIMCA) demonstrated that guaiacol-producing and non-guaiacol producing Alicyclobacillus strains could be differentiated up to 89% of the time. This technique may provide a tool for fruit juice producers to detect Alicyclobacillus rapidly and to monitor and control guaiacol formation.

Beverages↗
PubMed2006

Microbial quality of commercial 'Golden Delicious' apples throughout production and shelf-life in Lleida (Catalonia, Spain).

'Golden Delicious' apples taken from different points throughout the production chain and shelf-life were analysed for Enterobacteriaceae, thermotolerant coliforms, E. coli, E. coli O157:H7 and Salmonella spp. A total of 36 samples picked up in the orchard, 36 after storage and handling in the packinghouses and 144 from different retail stores were analysed using standardised techniques. Enterobacteriaceae counts were very variable. Pantoea spp. was the main genera isolated. The percentage of samples with thermotolerant coliforms was 16.6%, 22.2% and 10.4% after harvest, after handling in the packinghouse and in the stores, respectively. Strains belonging to Citrobacter, Enterobacter, Klebsiella and Escherichia were isolated. Three samples coming from orchards, five from the packinghouse and two from retailers harboured E. coli. However, none of the E. coli strains isolated had the virulence genes that are pathogenic for humans. None of the samples was Salmonella positive. This is the first study regarding microbial safety of fresh apples in the European Union (EU).

Colony Count, Microbial↗
PubMed2005

Experimental evaluation of the Corneal Concept 360 intraocular lens with the Miyake-Apple view.

PURPOSE: To evaluate a new single-piece, hydrophilic acrylic intraocular lens (IOL) designed to keep the anterior capsule away from the IOL optic, create the effect of a capsular tension ring, and prevent posterior capsule opacification (PCO). SETTING: John A. Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. MATERIALS AND METHODS: Four Concept 360 IOLs (Corneal Laboratoire) were implanted in 4 human cadaver eyes of various sizes (Miyake-Apple technique). Lens centration, ovalization of the capsular bag and capsulorhexis opening, distance between the anterior optic surface and the anterior capsule and between the posterior optic surface and the posterior capsule, as well as presence of capsular striae were evaluated. The capsular bag was stained with trypan blue, and the ophthalmic viscosurgical device (OVD) was stained with fluorescein to enhance visualization. RESULTS: In each eye, the lens conformed to the capsular bag with precise centration. No ovalization or distortion of the capsular bag or the capsulorhexis and no posterior capsule striae were observed. Measurements of the capsulorhexis diameter before and after IOL implantation were the same. The anterior capsule remained at distance from the anterior optic surface for 360 degrees, and tight contact between the posterior optic edge and the posterior capsule was observed. CONCLUSIONS: Use of the Miyake-Apple technique is useful in the evaluation of new IOL designs in experimental studies. Design features of the Concept 360 IOL are likely to decrease the outcome of postoperative capsular bag opacification. Long-term clinical studies are necessary to confirm these findings.

Acrylic Resins↗
PubMed2005

Optimal timing of capsular tension ring implantation: Miyake-Apple video analysis.

PURPOSE: To evaluate the appropriate timing for capsular tension ring implantation in cases of zonular weakness either prior to or after lens extraction using Miyake-Apple video analysis. SETTING: John Moran Eye Center, Salt Lake City, Utah, USA. METHODS: Four cadaver eyes were prepared using a standard Miyake-Apple protocol with image capture using digital video recording. After continuous curvilinear capsulorhexis and hydrodissection/viscodissection were performed, 2 eyes had early capsular tension ring implantation (CTR) and 2 eyes had CTR implantation after lens extraction. The 12.3 mm CTR was implanted in all eyes. Capsular bag torque and displacement, zonular elongation and stress, and ease of CTR placement were evaluated in each eye. RESULTS: Early CTR implantation resulted in significantly increased capsular torque and displacement of up to 4.0 mm compared to insertion in an empty capsular bag. There was significant zonular elongation and tension during early placement. CONCLUSION: In terms of minimizing further zonular stress and damage and capsular destabilization, the ideal timing for CTR placement is after lens extraction and decompression of the capsular bag.

Biocompatible Materials↗
PubMed2006

Imposex in the golden apple snail Pomacea canaliculata in Taiwan.

The golden apple snail Pomacea canaliculata (Lamarck, 1822) was introduced into Taiwan intentionally in the early 1980s and has become a recurring pest that seriously threatens aquatic crops. In this study, a field description of imposex with a developed penis sheath and penis in female golden apple snails from crop/domestic wastewater drainage sites and a six-order river is presented for the first time. Based on the five field collections and the aquarium group, the vas deferens sequence (VDS) of P. canaliculata in imposex development was categorized into four stages, i.e., stage 0: without male genital system; stage 1: with rudimentary penis; stage 2: with rudimentary penis and penis sheath; and stage 3: the rudimentary penis developing into penis pouch and penis. The VDS indices varied between 1.07 and 2.82 and were lowest in the aquarium group and Yuanlin2. Regarding the severity of imposex, the aquarium group was less pronounced, as illustrated by the length of penis sheath and penis length, than the field collections (p<0.05). In respect of the penis length, males of the most imposex-affected site were up to 15% shorter than that of the aquarium group. Negative correlations between male penis length and female imposex characters (i.e., penis length and penis sheath length) were also observed.

Animals↗
PubMed1997

Dimers of a GFG hexasaccharide occur in apple fruit xyloglucan.

Apple fruit xyloglucan is predominantly built up from XXXG, XXFG, and XLFG units (G = beta-D-Glcp-, X = alpha-D-Xylp-(1-->6)-beta-D-Glcp-, L = beta-D-Galp-(1-->2)-alpha-D-Xylp-(1-->6)-beta-D-Glcp-, F = alpha-L-Fucp-(1-->2)-beta-D-Galp-(1-->2)-alpha-D-Xyl p-(1-->6)-beta-D-Glcp-). However, small amounts of oligosaccharides with a less heavily branched glucan backbone also occur. Structural analysis of two such oligosaccharides, isolated from a xyloglucan preparation digested with endoglucanase i.v., using a combination of FAB mass spectrometry and 1H NMR spectroscopy, afforded the identification of GFG and a dimer of GFG. The finding of the dodecasaccharide GFGGFG as a structural element of apple fruit xyloglucan is most unusual.

Carbohydrate Sequence↗
PubMed1999

The PAN module: the N-terminal domains of plasminogen and hepatocyte growth factor are homologous with the apple domains of the prekallikrein family and with a novel domain found in numerous nematode proteins.

Based on homology search and structure prediction methods we show that (1) the N-terminal N domains of members of the plasminogen/hepatocyte growth factor family, (2) the apple domains of the plasma prekallikrein/coagulation factor XI family, and (3) domains of various nematode proteins belong to the same module superfamily, hereafter referred to as the PAN module. The patterns of conserved residues correspond to secondary structural elements of the known three-dimensional structure of hepatocyte growth factor N domain, therefore we predict a similar fold for all members of this superfamily. Based on available functional informations on apple domains and N domains, it is clear that PAN modules have significant functional versatility, they fulfill diverse biological functions by mediating protein-protein or protein-carbohydrate interactions.

Amino Acid Sequence↗
PubMed2000

Determination of fumaric acid in apple juice by on-line coupled capillary isotachophoresis-capillary zone electrophoresis with UV detection.

An on-line coupled capillary isotachophoresis-capillary zone electrophoresis (cITP-CZE) method for the determination of the fumaric acid content in apple juice is presented. A clear separation of fumaric acid in real samples is achieved within 20 min. The leading, terminating and background electrolyte of the employed system consist of 10 mM HCl+beta-alanine+5 mM beta-cyclodextrin+0.05% hydroxypropylmethylcelullose (HPMC), pH 3, 10 mM citric acid and 20 mM citric acid+beta-alanine+5 mM beta-cyclodextrin+0.1% HPMC, pH 3.3, respectively. The linearity, recovery, repeatability and detection limit of the developed method are 25-1000 ng/ml, 1.07%, 95.4+/-3.5 (+/-s)% and 10 ng/ml, respectively. Low laboriousness (no sample pretreatment), sufficient sensitivity and low running cost are the important attributes of the cITP-CZE method which was successfully applied to analyses of real samples of apple juices.

Beverages↗
PubMed2000

Determination of patulin in apple juice by high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry.

An HPLC-MS-MS method with selected reaction monitoring (SRM) for the determination of patulin in apple juice samples is described. Mass spectrometric detection was accomplished following atmospheric pressure chemical ionization (APCI) in both positive and negative ion modes. Collision induced dissociation (CID) of the protonated molecular ion led initially to the loss of H2O (fragment m/z 137). At higher energies CO is lost from both the protonated parent molecule (fragment m/z 127) and the dehydrated molecular ion (fragment m/z 109). In contrast, CID of the deprotonated molecular ion led initially to the fragment at m/z 109 corresponding to the loss of either CO2 or acetaldehyde, followed at higher CID energy by the loss of H2O (fragment m/z 135) and CO (fragment m/z 125) from the deprotonated molecular ion. Detection in the negative ion mode proved superior and a linear response was observed over the injected range from 6 to 200 ng patulin. Apple juice samples spiked with patulin between 10 and 135 microg/l were analyzed following liquid-liquid extraction with ethyl acetate and clean up with sodium carbonate. Utilizing reversed-phase HPLC with acetonitrile-water (10:90) at 0.5 ml/min, levels down to 10 microg/l were readily quantified and a detection limit of 4 microg/l was attainable at a signal-to-noise (SIN) ratio of 4. The MS data for the spiked samples compared well to the UV data and when plotted against each other displayed a correlation coefficient (R) of 0.99.

Atmospheric Pressure↗
PubMed2001

Separation of apple procyanidins into different degrees of polymerization by high-speed counter-current chromatography.

Apple procyanidins were fractionated by high-speed counter-current chromatography in a one-step operation from apple condensed tannins using a type-J multilayer coil planet centrifuge. The separation of procyanidins was performed with a two-phase solvent system composed of methyl acetate-water (1:1) by eluting the upper phase at a flow-rate of 1.0 ml/min. Each fraction was examined by time-of-flight mass spectrometry. Procyanidins were separated according to their degrees of polymerization.

Biflavonoids↗
PubMed1998

Incidence of patulin in apple juice concentrates produced in Turkey.

A total of 215 apple juice concentrate samples from three different producers in Turkey were analyzed for patulin using reversed-phase high-performance liquid chromatography. The detection limit of patulin in single strength apple juice at a sugar content of 11.2 degrees Brix was lower than 5 micrograms/l. Patulin was detected in all of the samples analyzed, at concentrations ranging from 7 to 375 micrograms/l. Of the samples, 43.5% were found to exceed a patulin contamination level of 50 microgram/l.

Beverages↗
PubMed1999

Simultaneous determination of 5-hydroxymethylfurfural and patulin in apple juice by reversed-phase liquid chromatography.

A rapid, simple and economical method was described for the simultaneous determination of 5-hydroxymethylfurfural (HMF) and patulin in apple juice. The sample was extracted with ethyl acetate and the extract was then cleaned up by extraction with a sodium carbonate solution. Then HMF and patulin were determined by reversed-phase liquid chromatography using a C18 column and a photodiode array detector. HMF and patulin could be completely resolved by using the mixture water-acetonitrile (99:1, v/v) as the mobile phase with a flow rate of 1.0 ml/min. Mean recoveries of HMF ranged from 86% to 100% with an overall mean of 94%, that of patulin ranged from 94% to 125% with an overall mean of 103%, for different spiking levels. The limits of detection for HMF and patulin in apple juice were found to be < 0.01 mg/l and < 5 micrograms/l, respectively.

Beverages↗
PubMed1987

Apple peel jejunal atresia.

Apple peel deformity of the small bowel is a variant of jejunal atresia with a high mortality. Forty five percent of these patients can be expected to die, most from anastomotic leaks with sepsis, or anastomotic failure with prolonged ileus and malnutrition. This report documents survival of three consecutive patients treated by an adaptation of the Santulli "chimney" anastomosis. This method accomplishes the goals of relief of intestinal obstruction, minimal resection, and preservation of tenuous intestinal blood vessels. All three patients had a benign postoperative course and made an uneventful recovery. Intestinal function is normal in each patient at 1 1/2 years. Based on this experience, it appears that a Santulli jejunojejunostomy combined with total parenteral nutrition offers the most promising operative strategy in the treatment of patients with apple peel bowel.

Female↗
PubMed2003

Formation of novel flavonoids in apple (Malusxdomestica) treated with the 2-oxoglutarate-dependent dioxygenase inhibitor prohexadione-Ca.

Novel flavonoids were formed in young leaves of apple (Malusxdomestica) after treatment with the dioxygenase inhibitor prohexadione-Ca, which is known to reduce the incidence and severity of fire blight caused by Erwinia amylovora and other plant diseases. The compounds were isolated and identified as luteoliflavan, luteoliflavan 5-glucoside, eriodictyol 7-glucoside and 6"-O-trans-p-coumaroyleriodictyol 3'-glucoside. These flavonoids represent a novel biosynthetic pathway in apple leading to the formation of 3-deoxyflavans. Concomitantly, the content of regularly occurring phenylpropanoids is also influenced by prohexadione-Ca with increasing amounts of hydroxycinnamic acids and decreasing flavan-3-ols and flavonols. The altered flavonoid metabolism may be related to the lowered pathogen incidence though the isolated novel flavonoids do not exhibit antibacterial activity.

Chlorogenic Acid↗