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Identification of a binding site for glycoprotein Ibalpha in the Apple 3 domain of factor XI.

Factor XI (FXI) is a homodimeric plasma zymogen that is cleaved at two internal Arg(369)-Ile(370) bonds by thrombin, factor XIIa, or factor XIa. FXI circulates as a complex with the glycoprotein high molecular weight kininogen (HK). FXI binds to specific sites (K(d) = approximately 10 nM, B(max) = approximately 1,500/platelet) on the surface of stimulated platelets, where it is efficiently activated by thrombin. The FXI Apple 3 (A3) domain mediates binding to platelets in the presence of HK and zinc ions (Zn(2+)) or prothrombin and calcium ions. The platelet glycoprotein (GP) Ib-IX-V complex is the receptor for FXI. Using surface plasmon resonance, we determined that FXI binds specifically to glycocalicin, the extracellular domain of GPIbalpha, in a Zn(2+)-dependent fashion (K(d) = approximately 52 nM). We now show that recombinant FXI A3 domain inhibits FXI inbinding to glycocalicin in the presence of Zn(2+), whereas the recombinant FXI A1, A2, or A4 domains have no effect. Experiments with full-length recombinant FXI mutants show that, in the presence of Zn(2+), glycocalicin binds FXI at a heparin-binding site in A3 (Lys(252) and Lys(253)) and not by amino acids previously shown to be required for platelet binding (Ser(248), Arg(250), Lys(255), Phe(260), and Gln(263)). However, binding in the presence of HK and Zn(2+) requires Ser(248), Arg(250), Lys(255), Phe(260), and GLn(263) and not Lys(252) and Lys(253). Thus, binding of FXI to GPIbalpha is mediated by amino acids in the A3 domain in the presence or absence of HK. This interaction is important for the initiation of the consolidation phase of blood coagulation and the generation of thrombin at sites of platelet thrombus formation.

Binding Sites↗

The use of a multimodal image, the apple technique, to facilitate clinical hypnosis: a brief communication.

A 1- to 3-minute exercise involving imagination (of an apple) and ideomotor ideation (hand levitation) is a simple, benign technique that is useful for illustrating to patients the nature of imagery and hypnosis. It avoids power struggles and allows a reasonable approximation of the patient's capacity for imagery and hypnotic responsiveness, without emphasizing the use of a hypnotic procedure. When administered to 35 college students, the hand levitation component of this exercise correlated with the Stanford Hypnotic Susceptibility Scale, Form C (Weitzenhoffer & E. R. Hilgard, 1963) (r = .66, p less than .001) and with the Stanford Hypnotic Clinical Scale: Adult (Morgan & J. R. Hilgard, 1975, 1978/79) (r = .60, p less than .001).

Adult↗

Health of tree swallows (Tachycineta bicolor) nesting in pesticide-sprayed apple orchards in Ontario, Canada. II. Sex and thyroid hormone concentrations and testes development.

To investigate the effects of pesticides on wild birds, sex (17beta-estradiol; testosterone) and thyroid (triiodothyronine (T3) hormone concentrations, body mass, and testes mass were measured and the development of testes was evaluated in wild tree swallows (Tachycineta bicolor) nesting in four sprayed apple orchards and three nonsprayed sites in southern Ontario, Canada, in 1995-1996. In orchards, birds were exposed to asmany as 11 individual spray events and five sprays of mixtures of chemicals. Residues of organochlorine pesticides, PCBs, lead, and arsenic concentrations were low and not variable among sites except p,p'-DDE concentrations, which ranged from 0.36 to 2.23 microg/g wet weight in eggs. These persistent compounds were not correlated with any endocrine response measured in tree swallows. In 16-d-old male tree swallow chicks, body mass and concentrations of 17beta-estradiol (estradiol), testosterone, and T3 in plasma showed no significant differences between sprayed and nonsprayed groups and among sites within those groups. However, T3 concentrations were slightly elevated in the sprayed group compared to the nonsprayed group, and there was a significant and positive correlation between T3 and the number of mixtures of sprays applied during egg incubation through chick rearing. In 16-d-old female chicks, there were no significant differences among spray treatments or sites and no correlations with spray exposure for testosterone, estradiol, or T3 in plasma. Body mass was correlated positively with T3 and negatively with estradiol but showed no differences among spray exposure groups or sites. Histology of testes of 16-d-old male chicks indicated there were no significant differences among sprayed and nonsprayed birds in testes mass, area, or diameter, or the presence of Leydig cells in the interstitium, the distribution of the Sertoli cells, or the occurrence of heterophils in the testicular interstitium. For the percentage of spermatogonia present on the basement membrane, there were significant differences among sites, but these differences were not specifically associated with spray exposure. However, there was a marginally significant trend between increasing occurrence of a disrupted Sertoli cell population on the seminiferous tubular basement membranes as the number of mixtures of pesticides sprayed during chick rearing increased. In adult male and female parent tree swallows, there were no differences in hormone concentrations between birds from sprayed and nonsprayed sites. Nor were there any significant correlations between the concentration of any hormone and collection date, body mass, or any type of spray exposure for adults. The correlations between increasing pesticide exposure and abnormal thyroid hormone and testes development in male chicks indicate that further reductions of pesticide use in orchards may benefit the health of birds that nest there. However, it is unclear which of these pesticides or spray mixtures are responsible for these effects, and this needs to be examined in future studies.

Animals↗

The reduction of patulin in apple juice by three different types of activated carbon.

Three types of activated carbon (NORIT SA 4, NORIT SX 4 and NORIT CA 1) were investigated for their ability to reduce patulin levels in apple juice at various Brix levels and temperatures. The steam-activated carbons (NORIT SA 4 and NORIT SX 4) exhibited similar adsorption isotherms at a dosage level of 1 g/l. They achieved patulin reductions of 80% and 70% respectively in 12(o) Brix juice at 55 degrees C. The similarity in performance between the steam-activated carbons implies that the purity and the surface acidity does not influence the adsorption of patulin. Chemically-activated carbon (NORIT CA 1) was less effective in removing patulin and achieved only a 45% reduction at a dose of 1 g/l. Patulin removal was influenced by juice Brix in that higher carbon doses were required at higher Brix levels for equivalent removal efficiency. At a dose of 1 g/l, NORIT SA 4 removed only 45% patulin from a 20(o) Brix juice. The removal of patulin from either 12 or 20(o) Brix juice by NORIT SA 4 at 1 g/l was not influenced by temperature changes in the range 30 to 65 degrees C.

Beverages↗

Epidemiology of mycotoxigenic fungi associated with Fusarium ear blight and apple blue mould: a review.

Research on the epidemiology of plant diseases is briefly reviewed, focusing on two diseases caused by mycotoxin-producing pathogens: Fusarium ear (or head) blight (FEB or FHB) of small grain cereals and apple blue mould (Penicillium expansum). Pathogen development during its key life cycles is discussed in relation to important environmental factors and host resistance. Current control methods are also reviewed, focusing on cultural and biological methods. The future challenge is to understand the relationships between disease severity, fungal biomass and the production of associated mycotoxins in order to minimize risks of both disease damage to crop yields and threat to human health posed by mycotoxins.

Food Microbiology↗

Comparison of HPTLC and HPLC procedures for the determination of certain xenobiotic residues in apples and pears.

HPTLC was used to check for residues of benomyl, carbendazim, ethoxyquin and thiabendazole in apples and pears. The method used showed good precision with a percentage coefficient of variation of less than 5%; recoveries were always greater than 90%. The limits of determination using HPTLC were always at least four times lower than Italian statutory limits. Selectivity with respect to other matrix components was excellent for all fruit varieties tested. Comparison with HPLC confirmed the validity of the results.

Benomyl↗

Effects of double encapsulation and coating on synthetic seed conversion in M.26 apple rootstock.

Encapsulated vitro-derived apical buds of M.26 apple rootstock (Malus pumila Mill) can be employed for the formation of the synthetic seed. Satisfactory levels of conversion (plantlets from synthetic seed) can be achieved if there are adequate (i) rooting induction treatment, (ii) protocol of encapsulation, and (iii) nutritive and environmental conditions. For capsule manufacturing, sodium alginate is largely used; however, this is excessively permeable with loss of the nutritive substances (artificial endosperm) and/or dehydration risks during conservation and transport causing detrimental effects on the synthetic seed conversion and on the plantlet's growth. In order to overcome these problems, two experiments were carried out comparing simple encapsulation in alginate with double encapsulation, and with encapsulation-coating procedures. The presence of a second layer of alginate (double encapsulation) and of a thin external coating layer over the alginate (encapsulation-coating) did not show any detrimental effects on viability, sprouting and regrowth of the encapsulated microcuttings. Satisfactory conversion (70%) was reached with the encapsulation-coating procedure, whereas the double and simple encapsulation converted less than 40% of the synthetic seed. The effect of the addition to the capsule of an anti-microbial substance (Plant Preservative Mixture - PPM) was examined: it did not compromise the conversion of the encapsulated microcuttings sown in ex-vitro non-aseptic conditions.

Drug Compounding↗

Behavior of parathion in apple juice processed into cider and vinegar.

Apple juice, fortified with 25 ppm (ug/g) of parathion, was processed into cider and vinegar. After the initial fermentation period of 12 days, the supernatant cider contained 7.4 ppm of parathion while the level in the sedimented lees was 88 ppm. Sorption to the sedimented matter was the main pathway for parathion residue reduction in the cider. Levels of aminoparathion and 4-nitrophenol, the only metabolites of parathion detected as confirmed by thin-layer chromatography, were 0.19 and 1.2 ppm, respectively, in the cider. The 56-day-old finished cider prior to bottling contained 2.2 ppm parathion, 0.15 ppm aminoparathion and 1.3 ppm 4-nitrophenol. Storage of the cider at 24, 12, 4 and -20 degrees C resulted in further reduction in the parathion levels. After one year, samples stored at 24 degrees C contained only 2.5% of the initial level added to the juice. Samples stored at the three other temperatures contained about 5% of the original level. Vinegar formed after 57 days of fermentation contained 5.1 ppm parathion, while the residue level in the lees was 76 ppm. Aminoparathion and 4-nitrophenol levels were 0.23 and 1.2 ppm, respectively in the vinegar. Storage of the vinegar at 24 degrees C for one year resulted in a gradual decline in the parathion level and at the end of the storage period, the remaining residue represented about 6% of the initial 24 ppm added to the juice.

Fermentation↗

Malabsorption of apple juice and pear nectar in infants and children: clinical implications.

Ingestion of fruit juices in which there is an imbalance between glucose and fructose (apple, pear nectar) and the presence of sorbitol is one of the major causes of the syndrome of chronic, nonspecific diarrhea of infancy and childhood. Stools in the condition at time of symptoms are acidic with a pH < 5.5, Clinitest positive, but free of blood and white cells. Lactose and sucrose breath hydrogen tests are normal but breath hydrogen is elevated if measured between 60-120 minutes after the offending juice is taken in sufficient quantity to elicit symptoms. Elimination of the offending juices immediately stopped the diarrhea in all of the cases. Three cases are presented documenting the variability of symptoms including one adolescent who became symptomatic while on a fruit and juice diet.

Beverages↗

Feeding and growth of apple snail Pomacea lineata in the Pantanal wetland, Brazil--a stable isotope approach.

Apple snails Pomacea lineata (SPIX 1827) are widespread in the tropical regions of Brazil as well as in the Pantanal wetland of Mato Grosso in the western part of the country. They have a key position in the Pantanal food web and serve as food for many animals e.g. fishes, birds, and caimans. However, little is known about their feeding preferences and growth rates. Stable isotopes have been used successfully on numerous studies as food source indicator. Therefore, the delta15N and delta13C values of snails from 0.45 to 3.03 cm in length, which were collected in the rainy season from March through May, were analyzed. Snails signatures revealed ambiguous evidence for food preferences. Delta15N and delta13C values ranged between -2.8 and 12.4 per thousand and between -24.2 and -16.4, per thousand respectively. This range of values mirrors the highly variable isotope values of possible food sources comprising C3 and C4 macrophytes. To test whether all common food sources were similarly assimilated, feeding experiments with different diets were conducted. Snail eggs were reared in tanks and offered different but single plants. Snails fed different diets and delta13C values of the food were reflected in the animal tissue. Growth varied considerably in experiments with different diets indicating the preference for certain food sources. Also, the fractionation of nitrogen isotopes between food and animal varied from 0.1 to 17.0 per thousand. The results are explained by different feeding habits, and it is supposed that animals fed either on the plant itself or on bacteria mats growing in the tanks. In an additional experiment juvenile snails were offered one single food with a distinctive C4 grass signature. These snails did not grow detectably, but nevertheless isotope signatures approached to values of the diet.

Animals↗

Residues and mutagenicity of captan applied to apple trees and potential human exposure.

The fungicide captan (cis-N-((trichloromethyl)thio) 4-cyclo-hexene-1,2-dicarboximide) was applied at the rate of 2.4 g/l to apple trees (c.v. Golden Delicious) individually or as part of a standard treatment program where it was applied eight times during the growing season together with several pesticides. Leaf samples (100 discs of 2.2 cm diameter) were collected from treated and control trees before treatment and at 0, 1, 3, 7, 14, 28, 56, 90 and 112 days after treatment. Fruit samples were taken at mid-season (56 days) and at harvest (112 days). The objective of this study was to determine the captan residue and mutagenicity of leaf and fruit extracts to ascertain the potential health hazard to agricultural workers in these orchards. Surface residues were extracted from leaves and fruits with methylene chloride. These extracts were subsequently analyzed for captan by gas-liquid chromatography (GLC) utilizing an electron-capture detector, and for mutagenicity with two strains (TA98 and TA100) of Salmonella typhimurium, with and without microsomal enzyme activation. Positive mutagenic effects were observed with strain TA100 at 0-14 days post spray, even with extracts from one leaf disc's surface (3.8 cm2) of the single treatment. Captan residues in these samples indicated a decline from 9.3 micrograms/cm2 at 0 days to 0.80 micrograms/cm2 at 14 days and a trace after 112 days. With the standard treatment, in which captan was incorporated eight times in the program starting at the 7-day interval, leaf extracts showed mutagenic activity at 7, 14, 28 and 90 days. Captan residues at these intervals were 11.4, 5.0, 4.1 and 3.4 micrograms/cm2, respectively. Fruit sample extracts of the standard spray were mutagenic to the tester strains TA100 and TA98 both at mid-season and at harvest. Residues of captan on fruits declined from 10.4 micrograms/cm2 at mid-season to 1.1 micrograms/cm2 at harvest. No mutagenic activity was detected with extracts from fruit samples from the single captan application.

Captan↗

Kinetics of inhibition of polyphenol oxidase mediated browning in apple juice by beta-cyclodextrin and L-ascorbate-2-triphosphate.

Polyphenol Oxidase (PPO) mediated browning in raw fruits and vegetables is a major cause of quality deterioration in fruits and vegetables and derived food products. Here the rate of browning reaction in apple juice treated individually and in combination (1:1) of beta-Cyclodextrin (beta-CD) and L-Ascorbate-2-triphosphate (L-AATP) is described. It was observed that the rate of quinone formation can be minimized using a combination of beta-CD and L-AATP as compared to individual treatment with these agents. Kinetic experiments revealed that both compounds are non-competitive inhibitors of PPO.

Ascorbic Acid↗

Glutathione and cinnamic acid: natural dietary components used in preventing the process of browning by inhibition of Polyphenol Oxidase in apple juice.

Consumer demands for 'freshness' in processed foods has been given increasing attention by food processing industries by searching for minimally processed products. Polyphenol Oxidase (PPO) mediated browning is a major cause of undesirable flavors and nutritional losses in fruit juices. Here the anti-browning efficiency of glutathione (GSH, reduced form) and cinnamic acid (CA) in apple juice is evaluated. It was observed that the rate of the browning reaction could be efficiently delayed using GSH and CA, which act as inhibitors of PPO. Kinetic studies confirm that GSH and CA are non-competitive and competitive inhibitors of PPO respectively.

Beverages↗

Antibody-based detection of acid-shocked, acid-adapted, and apple juice-incubated Escherichia coli O157:H7.

Sandwich enzyme-linked immunosorbent assays (sELISA) allow for rapid detection of Escherichia coli (E. coli) O157:H7. Acidic conditions similar to those in certain foods and juices may reduce the ability to detect E. coli O157:H7. Growth of E. coli O157:H7 at pH 4 compared to pH 5-7 reduced fluorescent signal at the lower bacterial concentrations without altering the range of detection. Both acid-adaptation and a subsequent pH 7 incubation reversed sensitivity. Incubation in apple juice was not deleterious to sELISA detection. Exposure to acidic conditions can cause a small reduction in sELISA sensitivity used to detect E. coli O157:H7.

Acids↗

[Energetic utilization of sugar beets, dried shreds, sucrose and apple pectin by growing swine. 1. Study overview and composition and digestibility of feed and rations].

Total metabolism experiments with a basic ration were carried out over two measuring periods and with a basic ration plus supplements of raw sugar beets, steamed sugar beets, dried sugar beet pulp, sucrose and apple pectin resp. with 4 pigs per test group and 4 test groups (LW 40-135 kg). Live weight gain was approximately equal after the feeding of raw and steamed sugar beets. It amounted to 650 g/animal and day in the first experimental period and to 715 g in the second. After feeding dried sugar beet pulp as supplement the corresponding weight gain was 525 and 565 g/animal and day. In the course of the ontogenetic development up to a live weight of about 70 kg an increase of digestibility could be observed. Adaptation cannot be excluded. This question will be pursued in the following experiments, so that consequences for energetic feed evaluation have to be postponed. For both raw and steamed sugar beets the digestibility of the organic matter was 88%, of NFE 95% and of water soluble carbohydrates 95%.

Animal Feed↗

[The energetic utilization of rations with dried concentrates, potatoes, sugar beets, apple pectin and cane sugar by growing, intact and ileorectostomized swine. 1. Results in swine with ileorectal anastomoses].

Rations with dried feedstuffs and rations with between 30 and 50% raw potatoes in the dry matter as well as steamed potatoes and sugar beets, sucrose and apple pectin were tested in measurings of the total metabolism in growing pigs with ileo-rectal anastomoses. The prececal digestibility of the energy of the rations ranged between 42% (rations with raw potatoes) and 79% (rations with sucrose). The digestibility of the rations with a 50% quota of steamed potatoes was better by 30%--units than that of rations with raw potatoes. In addition to crude nutrient digestibility, data are given on the digestibility of starch, water soluble carbohydrates and pectin. The wide variation in the N retention of 6-25 g/animal.day can only be partly connected with energy retention. Urine energy amounted to between 2.5 and 5% of gross energy. The average utilization of metabolizable energy amounted to 69.4%. A connection is to be seen with the value of energy maintenance requirement of 646 kJ retention energy/kg LW0.62.d.

Anastomosis, Surgical↗

[The energetic utilization of rations with dried concentrates, potatoes, sugar beets, apple pectin and cane sugar by growing, intact and ileorectostomized swine. 2. Comparison of results].

Results of the measuring of the total metabolism of growing intact pigs (INT) and pigs with ileorectal anastomoses (IRA) are compared. The 16 comparisons are based on studies with rations of dried feedstuffs and rations containing between 30 and 50% of the DM raw or steamed potatoes and sugar beets, sucrose and apple pectin. On an average of all comparisons the relative values for the digestibility of energy, DM and the organic matter as well as for the metabolizability of the energy were between 81 and 82% (values measured at INT animals = 100). The comparison also refers to the crude nutrients, the water-soluble carbohydrates, the starch and the pectin. As a tendency, the IRA utilized the metabolizable energy better than the INT animals. The energy maintenance requirement of the IRA animals was--by 15%--significantly higher than that of the INT animals. There is not yet an answer to the question in how far comparative studies of INT and IRA animals make verified statements with regard to the differences in the energetic utilization of the precaecally and postileally digested nutrients possible. For this, further experiments will be necessary.

Anastomosis, Surgical↗

Exploring within-tree architectural development of two apple tree cultivars over 6 years.

The present study addresses the prediction of apple tree development, taking into account both the number and within-tree position of tree components. The architectural development of two trees per scion cultivar, 'Fuji' and 'Braeburn', was studied by describing all shoots over 6 years. Flowering and fruiting were observed over 3 years. The description included different scales [entire trees, axes, growth units (GUs) and metamers], and the analysis compared all axes of the trees as a function of their branching order and age. Three main aspects of vegetative development were investigated: the quantity of primary growth; the number and nature of developing axillary shoots; and meristem death. Results confirm the existence of within-tree morphological gradients, and show that the decrease in growth was comparable in magnitude for all axes and GUs, irrespective of their position. This decrease results from a reduction in the number of metamers per GU, which was modelled by an exponential function. The decrease in growth involved changes in the number and nature of the axillary shoots, which could be described by simple functions. The probability of spur death was constant over the years but differed according to cultivar and type of bearing shoot. The within-tree probability of flowering and fruiting was predictable for 'Braeburn' because axes, regardless of their position and type, had a high probability of flowering and a low probability of fruit set which led to a regular bearing habit. In contrast, 'Fuji' had an alternating bearing behaviour that was more complex to predict. This appeared to result from a synchronized increase in the probability that all GUs at tree scale are floral, combined with a high probability of fruit set. The consequences of these results for both yield prediction and architectural simulations are discussed.

Flowers↗