FAV Health 2007 Poster Schedule
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101-P
A new approach to nutritive analysis using watermelon as a case study
Jennifer Waters* and Stephen King
Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, Texas A&M University, College Station, TX, USA, jennwaters@tamu.edu

Dietary strategies for disease prevention are most often viewed in discrete units such that only single nutrients associated with disease risk reduction are assessed at a time. This approach ignores the benefits of whole food nutrition. For example, phenylalanine, an essential amino acid, has been shown to be associated with a reduced risk of Parkinson’s disease. One study measuring this effect showed significant risk reduction at a dose of 200mg/day. The USDA’s nutrient database lists watermelon as containing 46mg phenylalanine per 304g (2 cup) serving. This equates to 23% of the dose for Parkinson’s disease risk reduction from phenylalanine, if consumed daily. This is not, however, the limit of watermelon’s potential for disease prevention; there are six other phytochemicals in watermelon that have been associated with risk reduction for this disease, and when taken in total these phytochemicals may have an additive effect in reducing risk. This broader approach gives a more realistic and optimistic, view of whole foods potential for disease prevention. This presentation addresses 20 such cases, outlining a new kind of total benefit.

102-P
Extraction of mucilage from four cactus plants and evaluation of its prebiotic effect on the growth of 2 probiotic bacteria
Elhadi M. Yahia*, Aracely Anaya, Brenda Martinez, Marcela Lopez, and Atenea Lozano
Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, México, yahia@uaq.mx

Functional components such as dietary fiber, natural colorants, and antioxidants are some of the nutrients sought to include in the daily diet. Several plant species are promising sources of such components, such as the case of Opuntia spp. and other cactus species. Several species of cactus are growing in arid and semi-arid zones, most thrive well under high temperature, little water and poor soils, conditions unfavorable for the production of many other crops. Cactus plants serve as sources for fruit and vegetables, for medicinal and cosmetic purposes, as forage, as source for natural colors, among other uses. However, many of these applications are still very restricted to very few species and in very few countries, but in light of the global desertification and declining water sources, these plants are gaining more interest. Recently the amount of food and health supplements containing probiotics and prebiotics have increased considerably. Fibers have been used as the main source of prebiotics, and lactic bacteria have been widely used as probiotics in foods. Some cactus plants contain abundant mucilage, which is a complex carbohydrate, and in some plants such as nopal it is thought of as a possible ingredient for several applications such as thickening agents and natural additives. The objective of this study was to develop an adequate method for the extraction of mucilage from 4 types of cactus plants (prickly pear cactus stems, Aloe vera, mesquite, maguey), and to evaluate if their mucilage can act as a prebiotic promoting the growth of 2 lactic (probiotic) bacteria (Bifidobacterium lactis (BL) and Lactobacilli acidophilus (LA)). For the extraction of the mucilage from maguey, nopal and zabila, the tissue was homogenized, centrifuged at 10,000 rpm for 10 minutes and extracted with acetone, acetonitrile or ethanol, and the mucilage was precipitated and freeze-dried. For mesquite the tissue was either macerated in an alkaline solution (0.5% NaOH) at 75ºC with constant agitation for 7 min and left in water for 24 to 48 hrs at 8ºC, or extracted in water at 80-90ºC for 2 hrs and then left in water at 8ºC for 48 hrs and then seeds removed from the water, or dried seed pods were ground and the flour was diluted in water at 95ºC and left in water at 8ºC for 24 hrs, and mucilage was precipitated with isopropyl alcohol (1:1.5). The highest yield of mucilage was obtained with ethanol in maguey, acetone in nopal, acetonitril in Aloe vera, and from dried and ground pods of mesquite. The bacteria were cultivated in 2 types of media: MRS broth and enriched MRS broth with 1% mucilage. After serial dilutions, both bacteria were cultivated in Petri dishes with Agar-MRS, and incubated at 32ºC for 24 hrs. Growth was evaluated by spectrophotometry and by total count of colonies, and amount of colonies presented in enriched media was higher compared with the regular media.

103-P
The correlation of some nutritional components in 8 fruits with their total antioxidant capacity measured with six different assays
R.D. Corral A.1, E.M. Yahia*2, A. Carrillo1, G. Gonzalez3
1Facultad de Quimico-Biologicas, Universidad Autonoma de Sinaloa, Culiacan, Sinaloa; 2Facultad de Ciencias Naturales, Universidad Autonoma de Queretaro, Juriquilla, Queretaro, Qro, 76230, Mexico; 3CIAD, Hermosillo, Sonora, yahia@uaq.mx

We have correlated the contents of total soluble phenolics (TSP), vitamin C, vitamin E, β-carotene and total carotenoids (TC) in eight fruits (avocado, black zapote, guava, mango, papaya, prickly pear fruit and cladodes and strawberry), with the antioxidant capacity of hydrophilic and lipophilic extracts measured by six different antioxidant assays: N,N-dimethyl-p-phenylendiamine assay (DMPD), 2,2’-diphenyl-1-picrylhydrazyl assay (DPPH), ferric ion reducing antioxidant power assay (FRAP), oxygen radical absorbance capacity assay (ORAC), trolox equivalent antioxidant capacity assay (TEAC) and total oxyradical scavenging capacity assay (TOSC). Guava had the highest TSP content, followed by black zapote, strawberry, prickly pear cladodes (nopal), avocado, prickly pear fruit, and papaya. Guava fruit had the highest levels of vitamin C (ascorbic and dehydroascorbic acid) content, whereas the lower levels were found in black zapote and nopal; the remaining fruits had a low vitamin C ranging from 38.5 to 104.1 mg/100 g FW. Ascorbic acid was the main contributor to the total vitamin C content (> 85%), with exception of “nopal” and avocado (26 and 58%, respectively). Evaluated fruits showed significant differences (p<0.05) in vitamin E (α- and γ-tocopherol) content, with the exception of mango and strawberry, without differences between them. The highest vitamin E content was in black zapote, followed by avocado, guava, nopal, mango, strawberry, papaya and prickly pear fruit. γ-tocopherol was detected in all, except prickly pear fruit. Mango had the highest content of β-carotene and TC, and prickly pear fruit, guava, and strawberry had the lowest content of TC, while β-carotene was not detected in strawberry and prickly pear fruit. In relation with antioxidant capacity, values of hydrophilic extracts ranged from 78.9 to 6230.6 µmol of trolox equivalents (TE)/100 g with DPPH assay; 798.4 to 15420.4 µmol TE/100 g with DMPD assay; 257.1 to 7482.9 µmol TE/100 g with FRAP assay; 330.4 to 8591 µmol TE/100 g with ORAC assay; 222.1 to 2243.8 µmol TE/100 g with TEAC assay; and from 2055.5 to 8235.6 µmol TE/100 g with TOSC assay. Lipophilic values ranged from 0 to 91.2 µmol TE/100 g with DPPH assay; 0 to 145.2 µmol TE/100 g with DMPD assay; 8.4 to 31.8 µmol TE/100 g with TEAC assay; and from 13.7 to 61 µmol TE/100 g with TOSC assay. In general, lipophilic values were <5% of the hydrophilic values except for a very few samples. The hydrophilic extract of guava had the major value when evaluated with DMPD, DPPH, FRAP, TEAC and TOSC assays; whereas with the ORAC assay, black zapote was the major one. On the other hand, the hydrophilic extracts of papaya and prickly pear fruit presented the lowest values of antioxidant activity with all used assays. The regression analysis revealed that the content of TPS and vitamin C were highly correlated with the antioxidant activity of hydrophilic extracts evaluated by the six assays. On the other hand, only the antioxidant activity of the lipophilic extracts, evaluated by the DMPD assay, was highly correlated with the content of β-carotene and TC.

104-P
Carotenoid content of five fruits and vegetables and their bioconversion to vitamin A measured by retinol accumulation in rat livers
G. K. Ramirez-Padilla1, E.M. Yahia*2 , A. Carrillo-Lopez1
1Facultad de Ciencias Químico-Biológicas, Universidad Autónoma de Sinaloa, Culiacan, Sinaloa, México; 2Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, Juriquilla, Querétaro, Qro., México, yahia@uaq.mx

Vitamin A deficiency is caused by a habitual diet that provides too little bioavailable vitamin A to meet physiologic needs. The major sources of vitamin A in the diet are preformed vitamin A, commonly found in foods of animal origin, and provitamin A carotenoids, found in yellow and orange-fleshed fruit and vegetables and in dark-green leafy vegetables. Of the approximately 600 carotenoids found in nature, only three are important precursors of vitamin A in humans: α-carotene, β-carotene, and β-cryptoxanthin. β-Carotene is the major provitamin A component of most carotenoid-containing foods. For practical purposes, the conversion of β-carotene to vitamin A (retinol) is considered as 6:1 and other provitamin A carotenoids as 12:1, however there are indications that these conversions might not be always correct for all types of foods. The present study was undertaken to analyze the carotenoid composition of five fruits and vegetables and to determine the bioconversion of their carotenoids to vitamin A, by monitoring the levels of retinol in the liver and plasma of Wistar rats fed with these products. Fresh samples of carrots, parsley, spinach, mangoes and papayas were partially characterized by evaluating their color, total soluble solids and humidity, and the edible portion was frozen with liquid nitrogen and freeze-dried. β-Carotene content was analyzed by High Performance Liquid Chromatography (HPLC) and total carotenoids by Spectrophotometry. From the analyzed products, carrots presented the highest content of β-carotene followed by parsley, spinach, mangoes and papayas, with 32.8, 19.6, 19.1, 2.6, and 1.2 mg/100g, respectively. Spinach had the highest content of total carotenoids followed by parsley, carrots, papaya and mango, with 60.7, 56.7, 40.9, 13.9 and 5.5 mg/100, respectively. For the biological experiment, 4-week-old male Wistar rats were used, and given a standard diet for two weeks as an adaptation period, followed by a vitamin A free diet for 12 weeks (depletion period). The rats were divided in groups and received the different treatments (each containing 100 µg β-carotene) in addition to their vitamin A-free diet, for 14 days. Liver and plasma retinol was analyzed by HPLC, and the bioconversión was based on “Retinol Accumulation Factor” (RAF), which was calculated by dividing the β-carotene (RAFβ) or total carotenoids (RAFTC) intake by the total retinol accumulated in the liver. RAFβ values were 1/5.998 for papayas, 1/6.506 for carrots, 1/6.956 for mangoes, 1/17.263 for spinach and 1/31.535 for parsley. RAFTC values were 1/118.544 for carrots, 1/173.528 for mangoes, 1/516.209 for spinach, 1/473.386 for parsley and 1/679.235 for papaya. There were no significant differences in plasma retinol between treatments. Therefore, carotenoids from papaya, carrots and mangoes are more bioavailable sources since they produced a better conversion to retinol, while green leafy vegetables were less effective in accumulating liver retinol.

105-P
Novel Vegetable Cultivars from TAMU- improving human health benefits, flavor and productivity
Kevin Crosby*, John Jifon, Kil Sun Yoo, and Daniel Leskovar
Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, Texas A&M University, k-crosby@tamu.edu

Texas A&M University and the Texas Agricultural Experiment Station have been developing improved vegetable cultivars for 75 years. While yield, appearance and disease resistance have always been emphasized, nutritional value and flavor have recently become priorities. This is due to the lack of information about genetic components of many important phytochemicals, and knowledge of strategies to enhance them. The Weslaco vegetable breeding program and the Vegetable and Fruit Improvement Center have been developing new genetic lines with enhanced levels of beneficial phytochemicals for the last ten years. These include peppers, melons, tomatoes, onions and carrots. ‘TAM Mild Habanero’, ‘TAM Ben Villalon’ green/red chile, ‘TAM Valley Hot’ cayenne, ‘TAM Dulcito’ sweet jalapeno, ‘TAM Tropic Bell’, ‘TAM Peproncini’, and ‘TAM Mild Jalapeño 2’ are new pepper cultivars with improved quality, yield, flavor and antioxidant concentrations. Experimental pepper lines have been developed with greatly enhanced (500-600%) levels of flavonoids and ascorbic acid, compared to popular commercial cultivars. These will be commercialized within the next few years. New melon cultivars include ‘Pacal’ orange casaba and ‘Chujuc’ cantaloupe, both with high levels of beta-carotene and sugars, along with excellent flavor. New tomato lines with enhanced vitamin C levels and virus resistance are also being developed for the fresh market. A genome map of melon, focusing on important quality traits such as beta-carotene and sugars has also been developed. The novel DNA markers are utilized to expedite selection and cultivar development. Quality traits and the genes which condition them are now more important than ever before in vegetable breeding programs. The VFIC and TAES will continue to develop improved cultivars with enhanced human health benefits.

106-P
Stability of oleuropein and other phenolic compounds during various extraction and processing methods of olive leaves
Nasir, S. A. Malik*, Marc Villalobose, Jim Brockington and Joe. M. Bradford
USDA-ARS, Weslaco, TX, USA, nmalik@weslaco.ars.usda.gov

Olive leaves contain potent polyphenols that are well known to have profound health benefits for variety of aliment including but not limited to cardiovascular diseases, hypertension, various types of cancer, various viral diseases such as HIV, and several bacterial diseases. For this reason various preparation of olive leaves are sold in health stores at fairly high prices. While all of the products were labeled leaf extracts, we found that oleuropein content (most important and abundant polyphenols of olive leaves) on per gram dry weight basis were similar or even less in some of these products compared to olive leaves dried in our laboratory. Some of these products contained less than 50% of the oleuropein content written on their label. We conducted a study to investigate degradation of oleuropein under different drying and processing methods to assess if some of the discrepancies might be due to processing methods. In addition attempts were made to determine if simpler extraction methods could be developed for individuals to use at home or for herbal industry to produce better or more concentrated products based on extracts. Our initial data suggest that oleuropein degrades drastically by freezing and thawing of leaves; nearly 80% loss in 15 minutes. While freeze drying caused no degradation of oleuropein, we found only minimal losses in oleuropein when leaves dried at room temperature (25°C) or in the greenhouse under the sun (40°C). Drying leaves at high temperatures in oven (60-80°C) resulted in drastic losses oleuropein; i.e., nearly 80% reduction. Simple extraction method that could be conveniently used by users at home were investigated and found quite effective in extracting reasonable amounts of oleuropein and other polyphenols.

107-P
Antimicrobial activities of guava products against foodborne pathogen, E. coli O157:H7
Guochen Yang*1, Salam A. Ibrahim2, and Zhongge (Cindy) Lu1
1Department of Natural Resources & Environmental Design, 2Department of Human Environment & Family Sciences, North Carolina A&T State University, Greensboro, NC, USA, yangg.@ncat.edu

Food safety directly related to human health, and has been a major concern for the food industry, and the public, especially since September 11, 2001. The Public Health Security and Bioterrorism Preparedness and Response Act of 2002 aimed to protect the Nation’s food supply and reduce the incidence of foodborne illness by committing to new approaches to ensure the food safety supplies with a strong emphasis toward elimination of foodborne pathogens in plant and food products. Guava is not only a rich source of vitamins, but also possesses many unique medicinal and antimicrobial properties due to its abundant content of phytochemicals. The research objective was to investigate antimicrobial effects of guava products on the survival and growth of seven Escherichia coli O157:H7 strains in liquid medium. These strains were maintained in BHI broth. Guava fruits were sliced into small pieces and blended using a blender. Guava juice and leaves were then extracted using three solvents, water, methanol and hexane. Fruit extracts were dissolved in 10 ml BHI broth tubes to make a fruit solution of 5% (w/v). E. coli O157:H7 was inoculated into fruit solutions at 2 log cfu/ml. After incubation at 37oC for 24 h, samples were serially diluted 10 folds. The proper diluent was spread-plated on TSA in duplicate. After incubation at 35oC for 24-36 h, viable cell counts were obtained. Experiments were repeated three times. Guava products significantly reduced survival and growth of the tested foodborne pathogen strains. Water extracts had the highest antimicrobial activity, followed by methanol and hexane. Data demonstrated that guava extracts are a potential antimicrobial agent to ensure food safety.

108-P
Handling and storage life effects on quality and health-beneficial compounds of ‘ready-to-eat’ edamame
Chlodys Johnstone and Douglas D. Archbold*
Department of Horticulture, University of Kentucky, Lexington, KY, USA, darchbold@uky.edu

‘Ready-to-eat’ edamame may have potential as a healthy snack food. For this to occur, the edamame may be blanched in the pod, frozen for storage and shipping, and thawed on the display shelf prior to purchase. This study examined the effects of a delay before blanching, as might occur during periods of high yield or in transit to a processor, and post-thawing shelf life duration on quality traits and health-beneficial compounds, including pod color, total antioxidant activity, and content of soluble sugars, phenolics, ascorbic acid, and isoflavones, of 2 edamame cultivars. Pods were blanched for 90 s immediately after harvest or after 3 days of 4ºC cold storage, frozen and stored at -20ºC in ziplock bags, then allowed to thaw in 4ºC cold storage and sampled after 3, 7, and 14 days. Pod color showed no general trends in L*a*b* color space values with blanching delay or shelf life duration, though cultivar differences were evident. Glucose levels in fresh edamame stored for 3 days before blanching generally decreased, but sucrose levels showed no consistent general trend. During post-thawing shelf life, glucose and sucrose levels declined over the first 7 days. Total antioxidant activity measured via the FRAP assay also declined during cold storage, but total phenolic, isoflavone and ascorbic acid content did not change appreciably. The results showed that both a delay before blanching and length of post-thawing cold storage may have effects on some but not all edamame quality traits and health-beneficial compounds, but that some cultivars may be better suited than others for ‘ready-to-eat’ use. Overall, blanched, frozen, then thawed ‘ready-to-eat’ edamame retained acceptable quality and most health-beneficial compounds for at least 7 days in cold storage.

109-P
Bone mineral content and breaking strength of female rats fed onion, tomato and broccoli separately or in combination
Changzheng Wang*, Lingyu Huang and Cecil Butler
Human Nutrition Program, Kentucky State University, Frankfort, KY, USA, changzheng.wang@kysu.edu

Epidemiological data suggest that vegetable consumption may be beneficial to bone health. The objective of this study was to determine the effects of vegetables on bone metabolism in growing rats. Powders of freeze-dried onion, broccoli and tomato replaced 5% or 10% of corn starch in the control diet (AIN93G) to form the treatment diets. Eighty weaning female Fischer 344 rats were randomly assigned into 8 groups of 10 rats each. The baseline group was killed at the beginning of the experiment. Group A was fed the control diet (AIN93M); Group B, C, D, E, F, G were fed the control diet with 10% Onion, 10% broccoli, 10% tomato, 5% onion + 5% broccoli, 5% onion + 5% tomato, 5% broccoli + 5% tomato, respectively. The experiment lasted for 10 weeks before the rats were sacrificed. Left femur was cleaned of soft tissues, wrapped with saline soaked cheesecloth and kept in air-tight plastic bag at -80oC until analyzed. The external dimensions of the fresh bones will be measured by a precision caliper. Bone volume was determined by the difference between the weight of the fresh bone and its weight in water. BMC and BMD were determined by a dual energy X-ray densitometer (DEXA) (Model DPX-IQ, Lunar Corp). Bone breaking strength was determined by a three-point bending test. Bones were ashed at 550oC for 48 hr. There were no significant differences among the treatment groups in bone mineral content, bone density and breaking strength, even though the base-line group was the lowest in all the parameters studied. No interaction between the different vegetables was observed. These results suggest that the selected vegetables did not adversely affect the bone status of growing rats fed a diet adequate in calcium.

110-P
Foliar potassium fertilization of melons to improve marketable and human health quality attributes
John Jifon*1 and Gene Lester2
1Vegetable and Fruit Improvement Center-Texas A&M University, Texas Agricultural Experiment Station, Weslaco, TX, USA; 2USDA-ARS Subtropical Research Center, Weslaco, TX, USA, jljifon@ag.tamu.edu

Numerous potassium (K)-mediated processes such as catalysis, osmo-regulation and assimilate transport are key determinants of fruit quality properties such as sweetness, aroma, and phytochemical content. However, during fruit development, soil-derived K is seldom adequate to satisfy these processes. Supplemental foliar K application under controlled conditions has been shown to mitigate this apparent deficiency however, the suitability of potential K salts as foliar sources is still uncertain. In this study, the effects of six foliar K sources (potassium chloride - KCl, potassium nitrate - KNO3, monopotassium phosphate - MKP, potassium sulfate - K2SO4, potassium thiosulfate - KTS , and a glycine amino acid-complexed K, Potassium Metalosate, KM) on field-grown muskmelon (Cucumis melo L.) fruit quality parameters were evaluated. Weekly foliar K treatments were established starting at fruit set and continuing to fruit maturity. Tissue K concentrations, soluble solids concentrations (SSC), total sugars, and the human wellness compounds - ascorbic acid and β-carotene were generally higher in plants receiving supplemental foliar K than in the control plants. Significant differences were observed among K sources, with foliar KNO3 fertilization consistently resulting in poor fruit quality compared to fruit treated with the other K sources. Although there were no consistent trends among the other foliar K sources, the data are consistent with previous controlled environment research findings that supplementing soil K supply with foliar K applications during fruit development and maturation can improve the marketable and human health quality attributes of muskmelon fruit.

111-P
Carotenoids from ‘Ataulfo’ mango: composition, bioavailability and bioconversion to vitamin A
J. de J. Ornelas-Paz1, E.M. Yahia*2, A. Gardea1, M. L. Faillac3
1Centro de Investigación en Alimentación y Desarrollo, Hermosillo, Sonora, México, 2Facultad de Ciencias naturales, Universidad Autónoma de Querétaro, Avenida de las Ciencias s/n, Juriquilla, 76230, Querétaro, México, 3Department of Human Nutrition, Ohio State University, 325 Campbell Hall, 1787 Neil Av., Columbus, OH, USA, yahia@uaq.mx

‘Ataulfo’ mango (Mangifera indica L.) is one of the most produced and consumed mango in Mexico, and also becoming important in other market such as USA. It is assumed that ‘Ataulfo’, as other mango cultivars, is a good source of dietary β-carotene, however its actual β-carotene composition has not been determined yet. In the present work, the content of β-carotene was measured by HPLC during the ripening process and was related with the internal and external color values. The lowest and highest content of β-carotene were 0.25 and 3.97 mg/100 g, respectively. Equations, with high determination coefficients (R2= 0.86-0.94) to predict the content of β-carotene from ‘Ataulfo’ mango on the basis of its internal and external color values were obtained. In other experiment, the pulp of ´Ataulfo´ mango at several stages of ripening was subjected to simulated in vitro digestion in presence or absence of a chicken baby food (CBF), as a fat source, and the transference of β-carotene to the micellar fraction (MF) was followed by HPLC. Only a slight increase in micellar β-carotene was detected when fruit at increasing ripening stages were tested in the digestive reactions without fat. Fat addition increased up to 3.3 fold the transference of β-carotene to the MF. The MF from digestions of fully ripened fruit and CBF were used to test the uptake of micellarized β-carotene by monolayers of Caco-2 cells, which incorporated the 16.9 % of β-carotene contained in the test medium. Finally, Wistar male rats were depleted of vitamin A over 12 weeks and then were fed fresh carrot or ‘Ataulfo’ mango for 2 weeks. Both test meals provided the same amount of β-carotene during the repletion period. At the end of the experiment, the retinol accumulation in the liver was found to be higher in the group fed with ‘Ataulfo’ mango (1642.3 μg retinol / liver) than in the group fed with carrot (1285.1 μg retinol/liver), suggesting that β-carotene from ‘Ataulfo’ mango was more bioavailable than the β-carotene from carrot. The obtained results demonstrated that ‘Ataulfo’ mango possesses a high content of β-carotene, which is bioavailable and bioconverted to vitamin A.

112-P
The absorption efficiency of α-cryptoxanthin from carrot leaves and β-cryptoxanthin from papaya
E. M. Yahia*1, D. E. Breithaupt2, J. F. Valdes1
1Facultad de Ciencias Naturales, Universidad Autonoma de Querétaro, Avenida de las Ciencias s/n, Juriquilla, 76230, Querétaro, Mexico, yahia@uaq.mx
2University of Hohenheim, Institute of Food Chemistry, Garbenstrasse 28, 70599 Stuttgart, Germany

The carotenoids xanthophylls (such as lutein) and zeaxanthin have gained increasing interest in recent years because of their positive health effects, such as in the prevention of eye diseases, age-related macular degeneration and cataract formation. Unlike α- and β-carotene, lutein and zeaxanthin are not considered to be provitamin A active as they are not converted in the human body into retinol. More likely they are supposed to act by absorbing blue light which damages the retina and by enhancing the antioxidative status of the environmental tissues. These findings led to an expanding international market for xanthophyll supplements, especially for eye health formulations. Numerous human intervention studies have been conducted with lutein to estimate its bioavailability from different formulations, but only a few human intervention studies discussed possible differences in the absorption efficiency of lutein and zeaxanthin. As far as we know, there has been no study that directly compared the lutein and zeaxanthin plasma levels reached after consumption of a formulated product. Therefore, the objective of our study was to investigate the absorbance efficiency of the monohydroxylated counterparts of lutein and zeaxanthin; α- and β-cryptoxanthin. A corn-oil based diet comprising β-cryptoxanthin from papaya puree and alpha-cryptoxanthin from green carrot leaves was fed to female Wistar rats for eight consecutive days, at a rate of 17.3 nmol and 9.2 nmol per day, respectively. The identity of the xanthophylls in the supplement was confirmed by mass spectroscopy [LC-(APCI)MS] analyses, and xanthophylls present in liver and plasma samples were determined by high pressure liquid chromatography. The β-cryptoxanthin concentrations of rat livers of the treatment group were statistically distinguishable (P < 0.01) from those present in livers of the control group that were fed a basic diet. α-Cryptoxanthin, the second xanthophyll present in the supplement, was not found in the rat livers of the treatment group. Plasma samples were free of xanthophylls. This is the first report proving that β-cryptoxanthin has a higher absorption efficiency than α-cryptoxanthin in rats.

113-P
Impact of nitrogen and sulfur fertilization on the phytochemical concentration of watercress, Nasturtium officinal R. Br.
Thomas C. Barickman*, Dean A. Kopsell, and Carl E. Sams
The University of Tennessee, Department of Plant Sciences, Knoxville, TN, USA, tbarickm@utk.edu

Brassica vegetables are commercially recognized for their nutritive values and contain beneficial phytochemicals that aid in health maintenance and disease prevention. Watercress (Nisturtium officinal R. Br.) is a leafy Brassica vegetable that provides a significant source of carotenoids and glucosinolates. Lutein, the main carotenoid in watercress, is a lipid-soluble pigment recognized as active in the prevention of cancers, cardiovascular, and eye diseases. Gluconasturtiin, an aromatic glucosinolate found in high concentrations in watercress, is a sulfur-containing phytochemical that exhibits both antioxidant and anticarcinogenic activity upon hydrolysis to phenylethyl isothiocynate (PEITC). Our research objective was to determine the influence of nitrogen and sulfur fertilization on carotenoid and glucosinolate metabolism in hydroponically grown watercress. Plants were grown in a controlled environment in a randomized complete block design. A factorial arrangement with three nitrogen treatments (6, 56, and 106 mg/L) and three sulfur treatments (8, 16, and 32 mg/L) were utilized. Leaf and shoot tissues were harvested and freeze dried to constant weight. Total concentrations of carotenoids and glucosinolates were measured by HPLC analysis. Nitrogen was positively correlated to lutein concentration which ranged from 2.43 to 9.52 mg/100 g fresh weight. Sulfur availability did not influence lutein concentration. Both nitrogen and sulfur treatments were positively correlated to gluconasturtiin concentration which ranged from 1.49 to 19.55 umol/g of dry weight. The impact of nitrogen and sulfur concentration on biomass, chlorophylls, and other individual carotenoids and glucosinolates will be discussed.

114-P
Manchurian wild rice (Zizania latifolia) infected with Ustilago esculenta stimulates innate immune system, via activation of human beta diffensin-2 and interleukin-8.
Yukihiro Oritani*, Eisaku Nishimura, Hiroyuki Inagaki and Tatsuhiko Ito
Health Food Science Research Institute, Morinaga & Co., Ltd., Japan, y-oritani-je@morinaga.co.jp

Manchurian wild rice (Zizania latifolia) is a vegetable broadly eaten in Asian countries, especially in China. It is also known that the stem of Zizania latifolia becomes swollen when the plant is infected with the parasitic fungus, Ustilago esculenta on the Ustilaginaceae Family in the Class of Heterobasidiomycetes. People have been cultivating this plant and eating the swollen shoots for more than 400 years. Zizania latifolia is traditionally said that it has immunostimulating effects. But its biological effectiveness and pharmacological mechanism has not been proved yet. The immune system can be divided into two major categories: innate immunity and adaptive immunity. The innate immune system confers non-specific protection against a large number of pathogens, hence, serving as the first line of host defense mechanisms. The immune system utilizes signal form toll-like receptors (TLRs) that bind and recognize microbial components. Zizania latifolia includes microbial components from Ustilago esculenta. Therefore, in this study, we have focused on the immunological stimulation activity of Zizania latifolia. We adopted the expression of hBD-2 and IL-8 as indicators of innate immune activation. hBD-2 ,the peptide responsible for innate immunity, is known to play a pivotal role in front defense against bacterial infection and it is induced by microbial components in epithelia. IL-8, which is known as a chemokine, is also induced by microbial components. In this study, we investigated the expression of hBD-2 mRNA in human colon carcinoma cells (Caco-2), quantified by real-time quantitative PCR at 9 hours post-stimulation by heat treated Zizania latifolia. The Concentration of interleukin 8 (IL-8) in the supernatant of cell culture medium was quantified by ELISA at the same time. As a result, the expression of hBD-2 was about 5-fold higher than the negative control. IL-8 was also induced by Zizania latifolia after 9 hours from stimulation. Additionally, we are going to examine the effect of Zizania latifolia to NK activity in BALB/c mice in order to reveal availability and significance of Zizania latifolia as a functional food.

115-P
Effect of passion fruit on skin health
Yuko Matsui*1, Toshio Takahashi1, Masanori Kamei1, Yotaro Katagata2, and Tatsuhiko Ito1
1Morinaga & Co., Ltd., 2Hirosaki University, Japan, y-matsui-jd@morinaga.co.jp

Passifloraceae is a plant from the South-America tropical forest and the leaves, vines and flowers are used as medical herbs. Many functions related to human health such as anti-anxiety, anti-inflammatory, cough suppressant, sedative etc., are reported. On the other hand, the fruit of passifloraceae, which is known as passion fruit, is mainly eaten in their natural state or processed to tropical juice. Although there are many reports on the functions of passifloraceae herbs, there are not so many on the fruit. Passion fruit includes polyphenols, which are noticed as anti-oxidative agents, and vitamins and minerals, which are known to be effective to healthy skin. It is said that skin aging occurs genetically and also environmentally. The damage to skin from UV exposure, mental and environmental stress, eating habits etc., lead to skin pigmentation, wrinkles and even skin cancer. To prevent these damages, the attention to anti-aging food is increasing. By the reasons described above, we have focused on the effect of passion fruit on skin health. Passion fruit was divided into three parts; rind (PF-R), pulp (PF-P) and seed (PF-S), and each part was extracted with 80% ethanol. The amount of polyphenol and the SOD activity of each part of the passion fruit were measured. The polyphenol concentration was significantly higher in PF-S than in PF-R and PF-P, and the SOD activity of PF-S was also higher. This indicates that PF-S acts as a strong antioxidant. The addition of PF-S to the culture of melanoma cells resulted in inhibition of melanogenesis, and also the production of total soluble collagen was much increased when PF-S was added to the culture medium of fibroblast cells. PF-R and PF-P did not show either effects. The results suggest that the application of PF-S has possibility to contribute to healthy skin. We will be investigating the relationship between the effect of passion fruit and skin health, and see whether these effects are related to the concentration and/or the kinds of polyphenols in the fruit.

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Characterisation of composition of pomace obtained from scab-resistant apples cultivars
Monika Kosmala1, Michal Sojka1, Krzysztof Kolodziejczyk1, Boguslaw Krol1, Malgorzata Uczciwek1, Jaroslaw Markowski2, Witold Plocharski*2
1Institute of Chemical Technology of Food, Technical University of Lodz, Poland, 2Department of Storage and Processing, Research Institute of Pomology & Floriculture, Skierniewice, Poland, witold.plocharski@insad.pl

Apple pomaces obtained during clear and cloudy juices production from scab-resistant apples were evaluated as a source of phytocomponents and dietary fibre. Apples of 27 scab resistant cultivars, harvested at commercial maturity, were cold stored for a few weeks at normal atmosphere, sorted in order to obtain representative samples and processed. Two samples of mash were pressed without enzymation, other two samples were depectinised using Rohapect MA PLUS (100 g/t, 20oC, 1 h) (AB Enzymes) then pressed. Apple pomaces obtained after the juice production were immediately frozen and lyophilized. Duplicated samples of each pomace were analyzed for dry substance, titratable acidity, total dietary fibre (TDF), and sugars and polyphenols content by HPLC. Average titratable acidity for all cvs. was 2,1 g/100g d.w. expressed as malic acid (min. 1,3; max. 3,9). TDF in samples without enzyme treatment was 50,3 g/100g (range 27,4 - 67,3). TDF in samples treated with enzymes was 55,2 g/100g (44,3 - 71,6). Total sugars content in pomaces not treated with enzymes was 39,9 g/100g (18,2 - 68,1), including saccharose 11,8 (0,6 - 24,1); glucose 6,7 (1,3 - 21,7); fructose 19,9 (9,2 - 41,3); sorbitol 1,4 (0,5 - 4,2); the ratio of glucose to fructose was 0,32 (0,14 - 0,57). Average content of total sugars in pomaces treated with enzymes was 33,6 g/100g (17,1 - 56,5), including saccharose 10,1 (0,4 - 25,2); glucose 5,6 (1,2 - 11,8); fructose 16,6 (8,6 - 25,2); sorbitol 1,2 (0,5 - 3,6); the ratio of glucose to fructose was on average 0,33 (0,14 - 0,58). Average content of total polyphenols in pomaces treated with enzymes was 292,1 mg/100g d.w. (min. 131,3; max. 530,7); with procyanidin B2 22,6 (range 10,6 - 62,8); sum of flawan-3-ols 125,4 (57 - 362,7); chlorogenic acid 28,2 (7,4 - 71,7); phloridzin 32,8 (13,6 - 67,4); sum of quercetin glycosides 98,2 (33,7 - 190) and quercetin 0,01 (from 0 for most cultivars to 0,4), cyanidin-3-galactoside 7,4 (0 - 19,6). Pomaces after mash enzyme treatment from 6 cultivars selected for further studies (Ariwa, Early Freegold, Florina, Novamac, Melfree and Rajka) were characterized by low procyanidin B2, total procyanidins and chlorogenic acid content. Quercetin glycosides were close to or above average level for all 27 cultivars. Phloridzin was on average level for Ariwa, Novamac, Rajka, above average level for Florina and even higher for Early Freegold. Pomaces from 6 selected cultivars were characterized by above average TDF content (51,7 - 57,8%). All researched pomaces contained 210 - 290 mg of polyphenols in 100 g dry matter, including 90 - 120 mg of quercetin glycosides. Selected scab-resistant cultivars considered as a good source of tasty juice are also a valuable source of dietary fibre and polyphenols, especially quercetin glycosides. They are characterized by low content of hydroxycinnamic acids and flavan-3-ols, and moderate quantities of phloridzin as well.
Results were obtained within “ISAFRUIT” Project, financed by the European Union - Contract No. FP6-FOOD 016279-2

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Effect of an acute flavonoid-rich juice intervention on postprandial plasma antioxidant status
Chutamat Niwat*, Trevor W. George, Michael H. Gordon and Julie A. Lovegrove
Hugh Sinclair Unit of Human Nutrition, School of Chemistry, Food Biosciences and Pharmacy, The University of Reading, Whiteknights, PO Box 226, Reading, Berks, UK, RG6 6AP, c.niwat@reading.ac.uk

Epidemiological studies have provided evidence that a high consumption of fruit and vegetables is associated with a reduction in the risk of cardiovascular disease. One possible mechanism for the protective effect of fruits and vegetables is the antioxidant activity of several bioactive components. Besides vitamins, minerals and dietary fibers, fruit and vegetables contain several types of phytochemicals. Flavonoids are a group of phenolic compounds with strong antioxidant activity present in fruits, vegetables, and other plant foods which contribute to the reduction of risk of chronic diseases such as cardiovascular disease. However, very little is known about the amount of flavonoids absorbed from the diet and the reliability of plasma biomarkers. The aim of the present study was to investigate the pharmacokinetics of flavonoid uptake and the beneficial effect of an acute intervention with flavonoid-rich juices on markers of oxidative stress and cardiovascular risk factors.
A single-blind, randomized postprandial study was performed in a group of twenty-four healthy free-living volunteers with low habitual fruit and vegetable intake, including 20 males and 4 females, with an age range of 28 to 64 years. Volunteers were asked to consume a low flavonoid diet for 5 days prior to the study day and fast overnight the day before the study. The volunteers were randomly allocated to consume either 400 ml of flavonoid-rich juice, providing 164 mg flavonoid, or 400 ml of flavonoid-poor juice, which had the same sugar content. Blood and urine samples were taken at time intervals after the ingestion of the juice and throughout the study day for 8 hours. In addition, the subjects were asked to return to give blood and urine samples the following morning (24 hours) and again in the evening (32 hours). One month later, they were asked to repeat the process with the second type of juice.
Plasma antioxidant status was measured before and up to 8 hours after the fruit juice consumption by the ferric reducing antioxidant potential (FRAP) assay and the oxygen radical absorbance capacity (ORAC) assay. It was observed that plasma FRAP was highest 1 hour after ingestion of the flavonoid-rich juice (P = 0.001) and the area under curve (AUC) for the FRAP response increased significantly by 9.1% during the 4-hour period after ingestion (P = 0.001), although this was not observed after the flavonoid-poor juice consumption. In addition, the AUC from the ORAC assay was significantly higher after ingestion of the flavonoid-rich juice compared with the flavonoid-poor juice (P = 0.05).
This study provided evidence that acute ingestion of flavonoid-rich juice can significantly increase plasma antioxidant capacity. This may be one mechanism by which consumption of fruit juices may reduce the risk of cardiovascular disease.
Funding provided by Unilever Bestfoods and The Royal Thai Government are gratefully acknowledged.

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Biochemical and nutritional Characterization of three prickly pear species with different ripening behavior
Talia Hernandez-Perez, Janet Mejia-Centeno, Andres Cruz-Hernandez and Octavio Paredes-Lopez*
Centro de Investigacion y de Estudios Avanzados del Instituto Politecnico Nacional (CINVESTAV-IPN). PO Box 629. Irapuato, Gto. Mexico, oparedes@ira.cinvestav.mx

The prickly pear is the fruit of the genus Opuntia, which belongs to the Cactaceae family (1). Prickly pear shows different patterns, depending on the time required for full fruit development from anthesis to physiological maturity. Prickly pear is an interesting food; its pulp is rich in glucose, fructose, and pectin and its peel contains cellulose, calcium, and potassium. Its use as human food fortifies and enriches the daily diet of the people living in the arid and semi-arid zones where it its cultivated (2). The main objective of the present study was to analyze some of the biochemical, nutritional and nutraceutical characteristics of prickly pears from three Opuntia morphospecies with different ripening behavior: Naranjona, Blanca Cristalina and Esmeralda (early, early-intermediate and intermediate-late ripening, respectively). Material and Methods. Physicochemical changes were measured according to AOAC (3). Carotenoid extraction was made as described by Delgado-Vargas and Paredes-Lopez, 1996 (4). Total carotenes and xanthophylls were determined by measuring absorbance at 436 and 474 nm, respectively, with an spectrophotometer; calculations were made as described by AOAC (3). Total dietary fiber (TDF) was evaluated using a commercial kit (Sigma-Aldrich), with a combination of enzymatic and gravimetric methods. Enzyme activities (poligalacturonase, pectinmetilesterase, β-galactosidase, cellulase) were spectrophotometrically evaluated after incubation with an specific substrate. Results and Discussion. The physicochemical analysis showed a decreased in the soluble solids content, titratable acidity (citric acid) and pH during fruit ripening. Blanca Cristalina and Esmeralda showed an increase in the carotenoids content, while this diminished in Naranjona. Xanthophyll levels showed similar behavior that carotenes, but in lower concentrations. TDF reached a peak value for three samples at day 7 after harvest, and decreased thereafter. The cell wall enzymes evaluated showed particular behavior during the ripening of each morphospecies suggesting a fine biochemical control and not a clear relationship between fruit softening and enzyme activity. Besides, the cladodia analysis showed differences in the β-carotene, chlorophyll and lutein levels. The final concentration was higher than other vegetables. Conclusions. Our results showed that the physicochemical, nutritional and nutraceutical characteristics of the prickly pears changed through the ripening stages and are somewhat related to the enzyme activity involved in this process.

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Phenolics in peaches and plums preferentially suppress the growth of estrogen-independent MDA-MB-435 breast cancer cells
Giuliana Noratto*1, David Byrne1, Luis Cisneros-Zevallos1 and Weston Porter2
1Department of Horticultural Sciences, Texas A&M University, College Station, TX, USA, giuliananoratto@yahoo.es
2Vet Integrative Biosciences, Texas A&M University, College Station, TX, USA

Commercial varieties of peaches and plums have been reported to contain a mixture of phenolics that may exert anticancer activity. Many of these phenolic compounds have been shown to inhibit the proliferation of a number of cancer cell lines (flavonoids), to induce apoptosis and inhibit metastasis of highly metastatic breast carcinoma cells (procyanidins and proanthocyanidins), and to exhibit chemopreventive and chemotherapeutic activity in vivo and in vitro (anthocyanins). However there are differences in the amounts and distribution of these phenolics among commercial varieties. Our objective was to evaluate the cancer suppression activity of extracts from a commercial variety of yellow fleshed peach Rich Lady (RL) and of red fleshed plum Black Splendor (BS) that had been previously quantified for their antioxidant activity and level of phenolic compounds. Frozen fruits were blended with methanol and left at 4°C overnight. The extracts were dried under vacuum at 45°C. Methanol extracts were then fractionated into phenolic acids (F1), anthocyanins (F2), flavonoids (F3) and procyanidins (F4) rich fractions by solid phase extraction using C18 cartridges. Cell proliferation was evaluated from 3 to 7 days of cell culture using a cell counter. All extracts, dissolved in culture medium at various concentrations expressed as mg chlorogenic acid equivalent/L (mg/L) were tested for their growth-suppression activity on the estrogen-dependent MCF-7, the estrogen-independent MDA-MB-435 breast cancer cells and one non-cancerous breast cell line MCF-10A. The peach RL extract effectively inhibited the proliferation of the estrogen-independent MDA-MB-435 breast cancer cell line. The IC50 was ~30 mg/L for this cell line as compared to IC50 of ~120 mg/L and ~ 515 mg/L for non cancerous breast line MCF-10A and the estrogen dependent breast cancer line MCF-7 respectively. In general BS extracts were less effective although they still affected the MDA-MB-435 to a greater degree than the other breast cancer cell line or the normal breast cell line. Within the three cells lines tested, the order of sensitivity to RL and BS extracts follows MDA-MB-435 > MCF-10A > MCF-7. Among RL and BS fractions, F3 and F4 have shown the highest potency against MDA-MB-435 cell proliferation but their effect on the non-cancerous MCF-10A cells remain to be investigated. In conclusion, peaches and plums may be added to the family of plants with potential anti-tumor activities

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Chlorella protothecoides is a new generation dietary source of omega-3 fatty acids and carotenoids for human ocular health
Faisal Ahmed*, Prakash Bhosale, and Paul. S. Bernstein
Department of Ophthalmology, Moran Eye Center, University of Utah School of Medicine, USA, faisal.ahmed@utah.edu

Lipophilic nutritional factors such as omega-3 fatty acids and carotenoids exert several important biological effects that may protect against retinal degenerative diseases such as age-related macular degeneration (AMD). The upcoming Age-Related Eye Disease Study II (AREDS II) is specifically focusing on the potential protective roles of the macular carotenoids (lutein and zeaxanthin) and omega-3 fatty acids (EPA and DHA); however, there are no reports of a single natural source that supplies both carotenoids and omega-3 fatty acids in appreciable quantities. In this work we have explored the potential of Chlorella protothecoides, a food grade algae, as a sole source of lutein, DHA, and other fatty acids.
Freshwater algae Chlorella protothecoides CS 41 was cultivated in Woods Hole MBL ( 50 ml) medium in 250 ml Erlenmeyer flasks and incubated at 20±1oC under continuous shaking (180 rpm) in dark/light alternating cycles on a rotary shaker at 180 rpm for 240 h. The cell mass was harvested and analyzed for carotenoids and fatty acids by HPLC and GC coupled with in-line single quadrupole mass spectrometers (MS), respectively.
Chlorella protothecoides displayed specific growth rate (μmax) and maximum biomass of 0.25 h-1 and 10.68 g l-1 in 240 h. The production profile indicated that lutein was the major carotenoid (95% w/w) produced throughout fermentation. At the end of the fermentation run (240 h), the total lutein content was observed to be 8.14±0.6 mg l-1. Lipid analysis revealed that Chlorella protothecoides accumulates 4 fatty acids in strong concentrations of the C16 & C18 series: (16:0, 18:1, 18:2, 18:3). Most importantly, it displayed a considerable (15 -23 % w/w of total lipid) amount of DHA (ω-3; 22:6) accumulation.
Production of lipids and carotenoids by Chlorella protothecoides suggests that it could act as a new generation nutritionally beneficial dietary source of lutein and DHA for human ocular health.

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Phytochemical contents and antioxidant activity of red leaf lettuce compared with 3 varieties of soft fruits
Saran Waroonphan*, Paulina Garcia-Macias, Michael H. Gordon, and Julie A. Lovegrove
Hugh Sinclair Unit of Human Nutrition, School of Chemistry, Food Biosciences and Pharmacy , University of Reading, Whiteknights, PO Box 226, Reading, Berkshire, RG6 6AP, England, UK, s.waroonphan@reading.ac.uk

Epidemiological studies have noted a consistent correlation between intake of diets rich in fruits and vegetables and reduced risk of chronic disease including cardiovascular disease (CVD) and cancer. There is considerable evidence that the beneficial health effects of plant foods may come from their phytochemicals, bioactive non-nutritive compounds which are not classified as vitamins, and may be more effective as antioxidants (Scalbert et al., 2005). Oxidative modification of low density lipoprotein (LDL) has been recognised as an early stage in the development of atherosclerosis which leads to CVD (Hassimotto et al., 2005). An increasing number of studies of the antioxidant effect of phytochemicals in fruits and vegetables including the retardation of the susceptibility of LDL to oxidation both in vitro and ex vivo have been reported (Laponite et al., 2006).
The phytochemical content and antioxidant activity of red leaf lettuce (Lollo Rosso), blueberries (Blue crop), raspberries (Joan squire) and strawberries (Elsanta) grown under plastic films in polytunnels were investigated. Flavonoid composition was determined by HPLC, total phenolics by the Folin-Ciocalteu assay, anthocyanins by the pH differential method and antioxidant activity by both the oxygen radical absorbance capacity (ORAC) assay and the copper-catalyzed oxidation of LDL ex vivo. The total phenolic contents were 2.9, 1.9, 2.96 and 3.07 mg gallic acid equivalents (GAE)/g fresh weight for lettuce, raspberries, strawberries and blueberries respectively. However, there was a big variation in different leaves of the lettuce with values ranging from 0.7 to 6.2 mg GAE/g for green and red leaves respectively. Blueberries contained the highest amount of anthocyanins (709 µg/g) compared with 625 µg/g for the lettuce but for strawberries and raspberries the anthocyanin content was almost 4 times less than for blueberries. The anthocyanin concentration was much higher in the red leaves compared with the green leaves of the lettuce (970 µg /g and 24 µg /g respectively). The antioxidant activity assessed by the ORAC values was higher for lettuce than for raspberries, strawberries and blueberries. ORAC values for raspberries, strawberries and blueberries were 30%, 57% and 72% of the values for the lettuce respectively. Comparison of the increase in LDL oxidation induction time ex vivo for samples containing extracts from fruits and lettuce confirmed the high antioxidant activity of the lettuce with the values of raspberries, blueberries and strawberries being 27%, 40% and 47% of those for the lettuce respectively. The ORAC values for lettuce also increased from 31 µmol Trolox Equivalents (TE)/g for green leaves to 135 µmol TE/g for red leaves.
In conclusion, it can be stated that Lollo Rosso lettuce is a good source of phytochemicals, with higher antioxidant activity than common varieties of soft fruit. The phenolic compounds and antioxidant activity are concentrated in the red leaves of the lettuce. Blueberries contained the highest antioxidant components among the fruits.

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Who will produce high quality, nutritious vegetables in the future?
Patrick Lillard* and Stephen King
Department of Horticultural Sciences, Texas A&M University, College Station, TX, USA, ptlillard@tamu.edu

Eating a wide variety of fresh, healthy fruits and vegetables requires an adequate supply at a reasonable price. The future vegetable supply chain will require that we have enough producers that are well trained and understand how to efficiently meet this growing demand for healthy nutritious vegetables, but current treads do not reflect this. At the beginning of the twentieth century 41% of the population worked on the land, but by the end of the century that had declined to less than two percent. The average age of the current American farmer is 54.3, with 26 percent of farmers over 65. With the rural population migrating to urban areas, there might not be anyone to take up the farm. In many of Texas urban areas such as Austin, Dallas and Houston there is a high demand for fresh, locally grown produce, but not enough farmers to meet the demand. The urbanization of the late nineteen hundreds has also created a generation gap between our stewards of the land and the current generations. The purpose of this study was to determine farmers reasons for growing and the motivating factors behind farming. Personal interviews were conducted with local growers using a purposive network sampling technique to select different growers. Participants were interviewed to obtain information regarding age, size of operation, farming experience, and reasons for growing. Interviews were transcribed, field notes incorporated as observational data, and data from each interview coded individually. Data was then examined for reoccurring themes. Three themes that emerged were health, family, and quality. Results from this research may help in recruiting and educating our next generation of farmers.

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Apoptotic and antiproliferative properties of cranberry extracts in colon and breast tumor cells
Anne M. Liberty*, Catherine C. Neto, and Peter E. Hart
Department of Chemistry and Biochemistry, University of Massachusetts-Dartmouth, North Dartmouth, MA 02747 2.Department of Biology, University of Massachusetts-Dartmouth, North Dartmouth, MA, USA, g_aliberty@umassd.edu

Cranberry fruit of Early Black cultivar (Vaccinium macrocarpon) was fractionated chromatographically and the fractions were analyzed for flavonoid, anthocyanin and proanthocyanidin content. The effects of the flavonoid fractions and ursolic acid, an abundant triterpenoid in cranberry peel, were assessed in two models of colon cancer and one model of breast cancer. Clonogenic soft agar assays were used to determine the effect of these compounds on tumor colony formation in HCT-116, HT-29 and MCF-7 cells. A metabolic response assay using MTT and a cell viability assay using trypan blue were performed to assess the ability of the different extracts to inhibit tumor cell proliferation. TUNEL assays were performed to assess apoptotic response to the cranberry compounds at different concentrations. The proanthocyanidins inhibited tumor colony formation in HCT-116 and HT-29 cells in a dose-dependent manner, with greater effect on the HCT-116 cell line. Ursolic acid strongly inhibited tumor colony formation in both colon cell lines. These compounds also decreased proliferation in all three tumor cell lines with the HCT-116 cell line most strongly affected. Ursolic acid produced a marked increase in the percentage of cells undergoing apoptosis in all cell lines at higher concentrations, with the most resistant cell line being MCF10A, a normal epithelial breast cell line. Cranberry proanthocyanidins had a stronger apoptotic effect in HT-29 colon tumor cells than the other cell lines.

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Flavonols and anthocyanins of elderberry fruits (Sambucus nigra L.)
Robert Veberic*, Jerneja Jakopic and Franci Stampar
University of Ljubljana, Biotechnical faculty, Slovenia, robert.veberic@bf.uni-lj.si

Sambucus nigra L. is a species of elderberry native to most of Europe, northwest Africa and southwest Asia. Elderberry is a red- purple colored fruit, which is used in Europe to prepare candies, jams, jellies and beverages. The use of different plant parts is long known in the traditional folk medicine. The juice squeezed from elderberry fruits contains red-purple pigments. The pigments of elderberry are anthocyanins, which are present in high amounts. Anthocyanins belong to the group of phenolics, which are known that beside their contribution to colour of fruits they contribute also to the taste and nutritional properties of fruit. We have tried to evaluate the flavonols and anthocyanins of elderberry fruits with the help of HPLC system. Phenolics were extracted from five different elderberry cultivars that are commonly grown in orchards and were breed in Austria. It was shown that different cultivars significantly differ in the amount of analyzed phenolics.

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Effects of maté-tea (Ilex paraguariensis) on DNA repair in mice
Daniel Duarte da Conceição Miranda1, Demétrius Paiva Arçari1, José Pedrazzoli Jr1, Patrícia de Oliveira Carvalho1, Suzete Maria Cerutti1, Deborah H. Markowicz Bastos*2, Marcelo Lima Ribeiro1
1Clinical Pharmacology and Gastroenterolgy Unit, Laboratory of Microbiology and Molecular Biology, Av São Francisco de Assis, 218 - Bragança Paulista-SP, Brazil. Zip Code 12916-900
2Nutrition Department- School of Public Health - São Paulo University - Av. Dr. Arnaldo 715, Zip Code 01246-904, São Paulo- SP- Brazil, dmbastos@usp.br

Oxidative DNA damage is involved in various pathological states such as cancer, and the presence of bioactive substances in the diet, such as phenolic acids and derivatives, is expected to help preventing the illness development by diverse, not yet elucidated, mechanisms. The aim of this study was to evaluate the antioxidant activity of maté-tea as well as the ability to influence DNA repair in male Swiss mice. Forty animals were randomly assigned to 4 groups. The animals received three different doses of maté-tea aqueous extract: 0.5 g.kg-1 or 1.0 g.kg-1 or 2.0 g.kg-1 for sixty days. After intervention, the liver, kidney and bladder cells were isolated and the DNA damage induced by H2O2 was investigated by the Comet assay. The DNA repair process was also investigated for its potential to protect the cells from damage by the same methodology. Results demonstrated that regular ingestion of maté-tea increased the resistance of DNA against H2O2-induced DNA strand breaks and improved the DNA repair after H2O2 challenge in the liver cells, irrespective of the ingested dose. These results suggest that maté-tea could protect against DNA damage and enhance the DNA repair activity through inhibition of DNA methylation and up-regulation of antioxidant enzymes. Protection might be afforded, also, by the free radical scavenging activity of the phenolic compounds present in this product.

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Antioxidant potential of fruits and vegetables commonly consumed in Ireland
Ankit Patras*1, Gwenole Gernigon1, Nigel Brunton1 and Francis Butler2
1Teagasc, Ashtown Food Research Centre, Ashtown, Dublin 15, Ireland, 2School of Agriculture, Food Science and Veterinary Medicine, University College Dublin, Dublin 4, Ireland, ankit_patras@rediffmail.com

Six vegetables, broccoli, carrot (organic and intensively grown) , butterhead lettuce, red onion, yellow onion and eight fruits, blueberries, cherry tomato, cranberries, red grapes, green pepper raspberry, red pepper, strawberry were analysed for anti-radical power (ARP), total phenols (TP), vitamin C (Vit C), total carotenoids (TC) and total anthocyanins (AC). The antiradical power (1/IC50) expressed as (g/l)-1 of the fruits ranged from 2.97 to 0.50 and 0.39 to 0.14 for vegetables. Red pepper and berries had the highest ARP values as compared to all the vegetables. Levels of phenols in vegetables varied between 375 and 158mg/100g DW and high levels of phenols (1430-385mg/100g DW) were detected in fruits while carotenoids were detected at very low levels in berry fruits (4.40- 0.64mg βCE/100gDW). Red pepper and carrots had the highest carotenoid content. Anthocyanins were found at very low levels in all the vegetables (0-5mg/100g DW), but highest anthocyanin content was observed in blueberries and cranberries. Berries showed the highest anthocyanin percentage among their phenols (17% from cranberry to 53% from blueberry). The ascorbic acid content of the fruits and vegetables as measured by the DCIP method ranged from 16 and 1975mg/100g DW. Principal Component Analysis was performed on the entire sample set of fruits and vegetables and the 2 first factors summed up 84% of the variability of the samples. PCA plot revealed that all the vegetables were located in the lower part of the first factor, which represents the antiradical power and confirmed that vegetables tend to have low antioxidant capacity (ARP). A strong correlation between antioxidant capacity and total phenols (ARP, r = 0.93) was found but ascorbic acid and total antioxidant capacity had a low correlation (r = 0.41). Carotenoids showed poor correlation with total antioxidant capacity (r = -0.13). Overall berry fruits had the highest antioxidant capacities. Antioxidant capacities of vegetables were lower, however, given their higher consumption levels they may make a significant contribution to overall antioxidant intake in the Irish diet.

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Nutraceutical compound concentrations of muscadine (Vitis rotundifolia Michx.) grape cultivars and breeding lines
Stephen Stringer1, Penny Perkins-Veazie2, and Donna Marshall*3
1USDA-ARS Thad Cochran Southern Horticultural Laboratory, 2USDA-ARS South Central Agricultural Research Laboratory, 3USDA-ARS Thad Cochran Southern Horticultural Laboratory, DAMarshall@msa-Stoneville.ars.usda.gov

Consumption of fresh fruits and vegetables has been linked both anecdotally and clinically to the prevention or alleviation of chronic diseases, and provides essential nourishment to mankind. The muscadine grape, a fruit indigenous to the southeastern U.S., contains several phenolic compounds that act as antioxidants and also other compounds, such as resveratrol, that can perform as a chemopreventative. The concentrations of these compounds present in the muscadine grape equal or exceed that known for other small fruit. Fruit of selected muscadine grape genotypes, including breeding lines and cultivars, were evaluated over a 2-year period to assess the existing genetic base for these nutraceutical compounds. Results demonstrated that concentrations of anthocyanins, total phenolics, ellagic acid, and resveratrol differ significantly among cultivars and breeding lines. These results suggest a possibility to breed for increased concentrations of the health-promoting compounds in muscadine grapes.

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Antioxidant activity of Castanea sativa Mill. extracts
Ibrahim O. Mujic*1, Jelena V. Zivkovic2, Zoran P. Zekovic3, Senka S. Vidovic3, Jasmina M. Tomin2, Vildana Alibabic4, Melisa Bajramovic4, and Slavica Dudas1
1Colegium fluminense Polytechnic of Rijeka Trpimirova 2/V,51000-Rijeka,Croatia, 2Medical Faculty of Nis, Department of Pharmacy, Bulevar Zorana Dindica 81,18000-Nis,Serbia, 3Faculty of Technology, Novi Sad, Bulevar Cara Lazara 1,21000 Novi Sad, Serbia, 4Faculty of Biotechnology, University in Bihac, Kulina bana 2, 77000 Bihac, Bosnia and Herzegovina, ibrahim.mujic@ri.t-com.hr

Antioxidant potential of sweet chestnut has been examined, as well as of Lovran chestnut and grafted Italian chestnut. Parts of chestnut fruit: fruit core, brown outer fruit rind, whole of the fruit, leaf, catkin, prickles and bark of chestnut tree have been extracted under the same conditions by 50% ethanol as extragens, afterwards the solvent has been completely removed by evaporating on vacuum-evaporator under lower pressure. In this way dry extracts of examined samples have been gained. Total phenolic content has been determined by Folin-Ciocalteu’s method and expressed as equivalent of gallic acid - GAE (mg GAE/100 g of sample). For determination of extract influence to creating and transformation of hydroxide (OH) radicals and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals, extracts have been dissolved in distillated water and added to Fenton\’s model system in concentration of 0.2 mg/ml. With the same concentration, the influence of commercial antioxidant, butylated hydroxyanisole (BHA), has been examined. Besides the chestnut tree bark with antiradical activity (AA) of 68.18%, high value AA in relation to hydroxyl radicals has turned out the brown outer bark of Lovran chestnut (59.36%), as well as catkins from Lovran chestnut (59.09%). The greatest antiradical activity in relation to DPPH radicals has shown the catkin of domestic sweet chestnut (37.50%) and then the extract of brown outer bark of Lovran chestnut with AA of 36.52%. Fruit core of chestnut didn’t show any antioxidant activity and antioxidant activities correlated well with the polyphenol concentrations.
Key words: Castanea sativa Mill., extracts, antioxidant activity, DPPH radical, OH radical, total phenolics

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Protocol development for presenting the hypoallergenic apple Santana to consumers with apple allergy
Marinus van der Maas*1, and Martijn Schenk2
1Applied Plant Research, Wageningen University and Research Centre, 2Plant Research International, Wageningen University and Research Centre, Rien.vandermaas@WUR.nl

The Dutch apple variety Santana was tested as the most hypoallergenic variety available for Dutch consumers. In cooperation with the Dutch Food Allergy Foundation and the Albert Heijn supermarketchain a protocol was developed to present Santana as a hypoallergenic apple to consumers. This protocol was tested in a pilot with 100 supermarketsbranches through conducting an inquiry. About 300 consumers responded to the inquiry. 41% of the consumers with apple allergy did not develop allergic reactions after eating Santana. Another group of 41% developed less allergy symptoms as usual and were satisfied with the product. The protocol was avaluated and adjustments were proposed.

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Research conducted at American universities on the nutritional value of sprouts
Emma Leigh Goodwin
Formerly, Sprouting Seeds etc. (distributer of seeds and equipment for sprouting seeds for food), pgoodwin7@aol.com

Research at U.S. Universities on the nutritional value of sprouted seeds in human nutrition started in earnest in the early 1940s in response to concerns about adequate amounts of protein in the diet during World War II. Dr. Clive McCay, a Professor of Nutrition at Cornell University in Ithaca, N.Y., wrote “Our daily paper would surprise us if it carried an ad: “Wanted: a vegetable that will grow in any climate, rivals meat in nutritive value, matures in 3 to 5 days, may be planted any day of the year, requires neither soil nor sunshine, rivals tomatoes in vitamin C, has no waste (in preparation). The Chinese discovered this vegetable centuries ago in sprouted soy beans. Today they are an important food for many millions.” About that time, a study of legume sprouts at Yale reported “...it appears that the common use of sprouts in the diets of Oriental peoples rests on a sound nutritional basis and should be introduced on a wide scale among Occidentals.” Since then, studies have been done at the universities of Idaho, Kentucky, Colorado State, Vermont, and Wisconsin. Of particular interest is one from the University of Kentucky: “This study shows that selected varieties of sprouted seeds may be a significant source of nutrients. The level of ascorbic acid is dramatically increased during germination of seeds. In addition, the concentration of protein, some B-vitamins, and minerals may make a considerable contribution to human dietaries when considered in light of their high nutrient/energy ratio in sprouted seeds.”
In the 1990s, Johns Hopkins Medical Institutions in Baltimore started conducting and publishing their work with broccoli sprouts and their ability to protect against human cancer. Indiana University followed, studying and publishing data on broccoli sprouts.

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Designing a molecular switch to optimize phenylpropanoid neutraceuticals in Vegetables
Meenu Vikram*, Yue Feng, Maria, Sunghun Park, Kil Sun Yoo and Hisashi Koiwa
Vegetable and Fruit Improvement Center, Departmet of Horticultural Science, Texas A& M university, College Station, TX, USA, mvikram@ag.tamu.edu

Phenylpropanoid, a plant derived organic compounds which includes flavonoid, lignin, coumarins and many small phenolic molecules, have a multiplicity of function in structural support, pigmentation, defense and signaling. Recent research suggest that dietary phenylpropanoid such as flavonoid may provide protection against cancer, aging, atherosclerosis, ischemic injury, inflammation and neurodegenerative diseases by contributing, along with antioxidant vitamins and enzymes, to the total antioxidant defense system of the human body. Arabidopsis PAP1 is a regulator in the biosynthesis pathway of phenylpropanoids that regulates the expression of multiple enzymes in the biosynthetic pathway. Overexpression of PAP1 in plants resulted in phenylpropanoid accumulation but caused retardation of vegetative growth. To optimize the level of phenylpropanoids accumulation in vegetables without compromising their growth, we designed a molecular switch that activates PAP1 via osmotic stress, thus the production of phytochemicals can be induced as pre- and/or post-harvest process. Arabidopsis cDNA encoding PAP1 and CBF3 were placed downstream of low-temperature and osmotic stress-inducible RD29a promoter. Since CBF3 is an activator of RD29a promoter, initial osmotic stress will result in induction of CBF3, which amplifies the transcription level of RD29a-PAP1 transgene. In order to test this system, RD29a-CBF3 and RD29a-PAP1 were co-transformed to Arabidopsis plants (WT and fry2 background, which is known to enhance the RD29a stress-inducible expression) through Agrobacterium mediated transformation. Transgenic plants were selected using Hygromycin (in MS media) and Basta (on soil) resistant genes as genetic markers. Molecular and phytochemical analyses are in progress using T3 double homozygous transgenic lines.

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Screening sea buckthorn for tocopherols and carotenoids
Staffan Andersson*1, Kimmo Rumpunen2, Eva Johansson3 & Marie Olsson1
1Dept. of Horticulture, 2Dept. of Plant Breeding and Biotechnology, 3Dept. of Agriculture, Swedish University of Agricultural Sciences, Sweden, Staffan.Andersson@ltj.slu.se

Sea buckthorn (Hippophae rhamnoides L.) berries are rich in different vitamins, antioxidants such as ascorbic acid, tocopherols and carotenoids, which are considered to exert beneficial health effects. Antioxidants may reduce the negative effects of free radicals and other reactive oxygen species which have the potential to damage cell components such as DNA, lipids and proteins. Some of the claimed health effects of sea buckthorn berries are reduction in plasma cholesterol level, inhibition of platelet aggregation, and regulation of immune function. Data about variation in amounts and types of vitamins and antioxidants in berries from different genotypes of sea buckthorn grown in the same location are scarce. In this study berries from 51 genotypes of sea buckthorn harvested in a germplasm collection were analyzed for composition and content of carotenoids and tocopherols. A large phenotypic variation in both amount and composition of carotenoids and tocopherols was revealed. This indicates a great potential to increase the content of these compounds in foods containing sea buckthorn berries, by proper choice of cultivars.

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Irrigation and nitrogen management to improve yield and nutritional quality of artichoke
Togo Shinohara, Shinsuke Agehara, Kil Sun Yoo, and Daniel I. Leskovar*
Vegetable and Fruit Improvement Center, Texas A&M University, Texas Agricultural Experiment Station, Uvalde, TX, USA, d-leskovar@tamu.edu

Artichoke (Cynara scolymus L.), a newly introduced crop in southwest Texas, is an excellent source of antioxidant phenolic compounds, including chlorogenic acid and cynarin. The edible part of the artichoke immature flower (head) is also known as a good source of minerals and dietary fibers. To investigate whether pre-harvest crop strategies can improve yield and nutritional quality of artichoke, a two-year field experiment was conducted with three irrigation regimes (50, 75 and 100 % crop evapotranspiration, ETc) and four N rates (0, 60, 120 and 180 kg N ha-1). Seedlings of artichoke cv. Imperial Star were transplanted in the 2005-2006 season, and off-shoots were allowed for re-growth in the 2006-2007 season. Differential irrigation was applied after stand establishment with a subsurface drip system. Nitrogen fertilizer was applied as NH4NO3 by side dressing in two equal split doses at stand establishment and head initiation each year. Yield, total head number, average head weight and size were highest at 100 % ETc, while 35% and 20% yield reduction occurred at 50 % ETc in the first and second season, respectively. Overall, nitrogen had small effects on head growth and yield. Total phenolics and chlorogenic acid content significantly increased with deficit irrigation, especially at late harvests. Cynarin content also showed similar trends, but they were not statistically significant. Conversely, soluble sugar and fiber content were similar among treatments but decreased as time of harvest progressed. Considering the sugar composition, the sucrose to monosaccharide (glucose and fructose) ratio increased during harvesting time. In this biennial production system, deficit irrigation strategies (lower than 100 % ETc) had a stronger impact than nitrogen rates on yield and the nutritional quality of artichoke heads.

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Flavor precursors and amino acids involved in blue-green pigment formation in crushed garlic cloves
Eun Jin Lee*1, Kil Sun Yoo1, Jung Eun Cho2, Seung Koo Lee2, and Bhimanagouda Patil1
1Vegetable and Fruit Improvement Center, Department of Horticultural Science, Texas A&M University, College Station, TX, USA, ejinlee@tamu.edu
2Department of Plant Science, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea

Blue-green pigments were occasionally formed in macerated garlic cloves after keeping it a while and that create some marketing problems in minced garlic products. In this study, we investigated garlic dicoloration by using mixtures of garlic and onion juices and isolated flavor precursors, trans-(+)-S-1-propenyl-L-cysteine sulfoxide (1-PeCSO, isoalliin) from onion and (+)-S-2-propenyl (or allyl)-(2-PeCSO, alliin) from garlic, respectively. The Pigment formation was associated with an alliinase activity and the increased levels of flavor precursors, particularly 1-PeCSO, and several amino acids. However, cysteine and proline did not result in coloration. Upon maceration of the tissue, 1-PeCSO and 2-PeCSO were enzymatically cleaved and yielded 1- and 2-propenyl-thiosulfinates. Subsequently, they reacted with amino acids and produced the pigments. Vivid-blue pigments were successfully produced by using a reaction system comprising only 1-PeCSO and 2-PeCSO, purified alliinase, and glycine (or some other amino acids). While pink pigments were generated from 1-PeCSO, yellow pigments produced from 2-PeCSO in a reaction of alliinase and amino acids, respectively.

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Separation of blue pigments in crushed garlic cloves: the color-forming potential of individual amino acids
Jung Eun Cho*1, Eun Jin Lee2, Kil Sun Yoo2, Seung Koo Lee1, and Bhimanagouda Patil2
1Department of Plant Science, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea, idmc@snu.ac.kr
2Vegetable and Fruit Improvement Center, Department of Horticultural Science, Texas A&M University, College Station, TX, USA

Unknown blue pigments, responsible for the discoloration in crushed garlic cloves, were separated and tentatively characterized by using a high-performance liquid chromatography (HPLC) and a diode array detector ranging at 200-700 nm. We developed several reaction systems for generating pure blue pigments using an isolated alliinase, isolated flavor precursors, trans-(+)-S-1-propenyl-L-cysteine sulfoxide (1-PeCSO) from onion and (+)-S-2-propenyl (or allyl)-L-cysteine sulfoxide (2-PeCSO) from garlic, and all 22 amino acids. Each amino acid was reacted with flavor precursors in alliinase solutions and then incubated at 40oC for several hours to generate pigments. At least 10 naturally occurring blue pigments were separated from discolored garlic extracts by using HPLC at 580 nm. This suggested that garlic discoloration was not caused by one blue pigment but many pigments. Also, all amino acids (except for cysteine, proline, and 4-hydroxy-proline) formed blue and purple pigments. Finally, we could demonstrate that the abilities of major amino acids to form the pigments in crushed garlic.

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Structural gene expression in carotenoid biosynthetic pathway of carrots during root development
Haejeen Bang*, Kil Sun Yoo, and Leonard M. Pike
Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, Texas A&M University, College Station, TX, USA, hbang@tamu.edu

Carrot root colors are mainly attributable to composition and content of carotenoids, which are health beneficial bioactive compounds for human. Carotenoid biosynthesis mechanism and root color inheritance are very complex in carrot, as compared to other crops. Major structural genes in the carotenoid biosynthetic pathway have already cloned and the sequence characterization has been reported. This study was carried to examine if differential carotenoid accumulation in various colored root were resulted from a mutation of structural genes at the transcriptional level. Roots were harvested at four different developmental stages in four different color carrots, yellow, orange, red and purple. A total of seven genes in the pathway were examined using RT-PCR. Regardless of different development stage, mRNA transcripts of all genes were detected but the level of expressions appeared to be differential during root development. It has not been confirmed whether carrot phytoene synthase1 (PSY1) is an ortholog of tomato PSY1 and carrot PSY2 is that of tomato PSY2 having same transcriptional behavior. However, the phylogenetic relationship and deduced amino acid similarity showed that carrot PSY2 was closely related to PSY1s in other crops, whereas carrot PSY1 was close to PSY2s. mRNA transcript of beta-carotene hydroxylase 2 was detected at reduced level compared to other structural genes in the pathway. Our results suggested that carotenoid accumulation in carrot may be regulated by regulatory genes rather than structural genes in the pathway.

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Influence of pre-storage hot treatment on lipid peroxidation, antioxidant enzymes and ntioxidant capacity of Persian limes (Citrus latifolia Tanaka)
Fernando Rivera-Cabrera*1, Janeth Jacuinde1, Fernando Diaz de Leon1, Clara Pelayo-Zaldívar2, Laura J. Perez-Flores1
1Department of Health Sciences, 2Department of Biotechnology, Universidad Autonoma Metropolitana, Mexico, D. F., 09340, Mexico, Ferivera2323@yahoo.com

The commercialization of fresh horticultural products and their processed by products is an important income source for countries with tropical and subtropical climates, like Mexico. Mexico is currently the world leading Persian lime producer and exporter, and quality standards have been developed in this country for this specie, which is the most recently exploited citrus fruit in Mexico. In general, plant cells produce reactive oxygen species (ROS) as part of their metabolism. ROS such as hydrogen peroxide, superoxide and hydroxyl radicals react with biomolecules including lipids, proteins and nucleic acids altering their function and leading to lipid peroxidation and eventually loss of membrane integrity, among other effects (Shewfelt and Purvis 1995; Mittler 2002). Increment of cell detoxification mechanisms can be a key factor against oxidative stress (Foyer et al. 1997). An increasing number of studies have shown that the induction of resistance to a severe stress can be caused by the exposure of the tissue to another moderate stress (Sabehat et al. 1998; McDonald et al. 1999; Schirra and Cohen 1999; Saltveit 2001). High temperature conditioning treatments can increase the antioxidant capacity of tissues and prevent the accumulation of ROS (Prasad, 1996). Furthermore, several reports have suggested that postharvest stresses enhance the nutraceutical content of fresh fruits and vegetables with a corresponding added value to the produce. In the present work, we evaluate the effect of a pre-storage hot water conditioning treatment (3 min 53C) on the lipid peroxidation levels, activity of peroxidase (PX) and superoxide dismutase (SOD) enzymes and antioxidant capacity in Persian limes (Citrus latifolia Tanaka) fruits stored at 25C and 80% RH. Heat conditioning treatment caused a significant increase in PX and SOD activities and lipid peroxidation levels in the fruits immediately after hot treatment. Preliminary results showed that the heat treatment did not have any effect on the total antioxidant capacity.

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Influence of ripeness on physical, chemical and antioxidant properties of blackberries (Rubus adenotrichus Schltdl.) cultivated in Costa Rica
Sonia Cozzano1, Óscar Acosta-Montoya1, Ana M. Pérez-Carvajal1, Floribeth Víquez1, Marco V. Castro2, Fabrice Vaillant*3
1Centro Nacional de Ciencia y Tecnología de Alimentos (CITA), Universidad de Costa Rica (UCR)
2Laboratorio de Tecnología Poscosecha, Centro de Investigaciones Agronómicas (CIA), Universidad de Costa Rica (UCR)
3Centre de Coopération Internationale en Recherche Agronomique pour le Développement (CIRAD-PERSYST), UMR QUALISUD, favillan@cita.ucr.ac.cr

Epidemiological studies have shown that diets rich in plant foods protect humans against degenerative diseases such as cancer and cardiovascular disease. Besides providing fiber, vitamins, phytosterols, sulfur compounds, carotenoids, and organic acids, plant foods contain a variety of polyphenols, which are increasingly regarded as effective protective agents. As with other fruits and vegetables, berries are important dietary sources of fiber and essential vitamins and minerals, and they also contain a vast number of other phytochemicals. Several authors have confirmed that blackberries are a good source of antioxidants, and are known to contain appreciable levels of phenolic compounds. Previous studies have reported the influence of ripeness on anthocyanins, polyphenols, and antioxidant properties in blackberry fruits. The aim of this study was to first evaluate the influence of maturity on physical, chemical, and antioxidant properties of a blackberry fruit grown in Costa Rica. Secondly, the correlation between the hydrophilic oxygen radical absorbance capacity (H-ORAC) values and bioactive compounds (total polyphenols, ascorbic acid, and total anthocyanins) in the assessed samples was evaluated. Organically grown blackberries (R. adenotrichus) were harvested from a 3-year old plantation located in Santa María de Dota, San José, Costa Rica. The berries were handpicked from 3 separate plots at the same plantation; plots were in a completely randomized design with about 30 plants per plot. Three composite samples (one from each plot) were picked for each one of the 3 stages of ripeness, based on fruit surface color, and were sorted as under-ripe (pink), half-ripe (part red-part black), and commercially ripe (full black). Whole fruits were analyzed for weight, size (length and width), compressibility and color. Homogenized pulp was analyzed for moisture, soluble solids, total titratable acidity and pH. Freeze-dried pulp was analyzed for ash, protein, dietary fiber, fat, total sugars, H-ORAC, total phenolic compounds, ascorbic acid and total anthocyanins. Fruits presented significant differences in weight, size, compressibility, color, moisture, soluble solids and protein for different developmental stages (P<0.05). Changes in ash, dietary fiber, carbohydrates, acidity and sugars were still significant but less marked. Ascorbic acid content did not vary significantly with ripeness and total polyphenols presented a somewhat erratic tendency. Anthocyanin content increased significantly with ripeness and the H-ORAC value for commercially ripe fruit was significantly higher than values for other samples. H-ORAC value for this stage of ripeness was 432±30 μmol TE/g (dry basis). H-ORAC values were positively and significantly correlated with total anthocyanin content (r = 0.98, P < 0.0001), but not with other components.

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Bioactive compounds content of yerba maté (Ilex paraguariensis) beverages
Deborah H. Markowicz Bastos*, Ana Claudia Fornari, Yara S. de Queiroz, Rosana Aparecida Manólio, Elizabeth A.F.S. Torres
Nutrition Department, School of Public Health, University of São Paulo, Av. Dr. Arnaldo, 715. Zip Code 01246-904 São Paulo, Brazil, dmbastos@usp.br

To conciliate low cost and well-accepted food items which may provide antioxidant substances in a regional diet is a challenge in public health. The availability of accurate and complete data on food composition is crucial to estimate the dietary intake of bioactive substances. Yerba maté (Ilex paraguariensis) is widely consumed in South America. It is rich in several bioactive compounds such as phenolic acids, which are absorbed by the body and may act as antioxidants or as free radical scavengers; caffeine, responsible for yerba maté stimulating properties long known by the native Indians in South America; and saponins, which may explain yerba maté popular use as both a choleretic and a digestant. The contents of caffeine and 5-caffeoylquinic acid (5-cqa) were determined in three yerba maté beverages: the chimarrão, widely consumed in the South of Brazil, Uruguay, Argentina and Paraguay; the tererê, consumed in the central west of Brazil and Paraguay and the mate-tea, consumed in the south-east of Brazil, Argentina and Uruguay. Both chimarrão and tererê are prepared with dried green maté leaves. Chimarrão is prepared with hot water and tererê is prepared with cold water. The maté tea is prepared with roasted leaves and brewed as any other herbal tea. The samples were purchased in two hypermarkets in São Paulo, Brazil. Four different brands of maté tea were purchased in boxes with 10 teabags each. Two different lots of each brand were analyzed. Three different brands of yerba maté, purchased in 1 kg paper bags, were analyzed. Analyses were performed by RP-HPLC using a diode array detector. Data were obtained at 272 nm for caffeine and 323 nm for phenolic acids. Identification was based on the comparison of the spectra obtained between 250-350 nm and the retention time of the unknown substances in relation to that of pure standards. Peak purity, which is determined using the average of similarity of the apex vs. the upslope spectrum and the apex vs. the download spectrum performed by the photodiode array detector, was higher than 0.99 for both compounds. Quantification was achieved by external calibration, using a five-point curve of different dilutions of a standard solution. One cuia (the apparatus used to drink chimarrão and tererê - 500 mL) of chimarrão contains 135 mg of caffeine and 226 mg of 5-cqa. One cuia of tererê contains 85 mg of caffeine and 163 mg of 5-cqa. One cup (182 mL) of mate-tea contains 13 mg of caffeine and 16 mg of 5-cqa. These data can be used to establish the dietary intake of bioactive compounds in these beverages.

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Maté tea (Ilex paraguariensis) ingestion effects on oxidative DNA damage and fatty acid profile in mice liver
Demétrius Paiva Arçari1, Fernanda Martins1, Fabiola Cristina Scagnolato1, Samara Bonesso dos Reis1, José Pedrazzoli Jr1, Deborah H. Markowicz Bastos*2, Suzete Maria Cerutti1, Patrícia de Oliveira Carvalho1, Marcelo Lima Ribeiro1
1São Francisco University - Av. São Francisco de Assis, 218. Jd. São José. Bragança Paulista. SP. Brazil
2Nutrition Department - School of Public Health - São Paulo University -Av. Dr. Arnaldo, 715 CEP 01246-904 São Paulo -SP-Brazil, dmbastos@usp.br

Currently, growing evidence suggests that plant-derived compounds may have beneficial effects on human health and that some even exert antioxidant activity. The aim of this study was to evaluate the effect of maté-tea (Ilex paraguariensis) on antioxidant activity and fatty acid composition in the liver of male Wistar mice. Forty animals were randomly assigned to 4 groups in accordance to the treatment and dose used. The animals received three different doses of roasted maté-tea aqueous extract: 0.5 g.kg-1 or 1.0 g.kg-1 or 2.0 g.kg-1 for sixty days. After treatment, the liver cells were isolated and the DNA damage and antioxidant activity induced by H2O2 were evaluated by the Comet assay. Fatty acid profiles were analyzed by high resolution gas-chromatography. Results demonstrate that maté-tea was able to decrease H2O2-induced DNA breakage in all groups. The fatty acid profile showed that the proportion of mono and polyunsaturated fatty acid was significantly higher in all treated groups. Conversely, the proportion of saturated fatty acid was significantly decreased in the treated groups. These results suggest that chronic treatment with mate tea for 60 days can protect the liver cells against DNA damage after H2O2-induced DNA breakage. Protection may be afforded by the antioxidant activity of the maté-tea´s bioactive compounds, such as phenolic acids, which can act as hydrogen or electron donors and as transition metal ions chelators.

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Bioguided fractionation of Uva caimarona (Pourouma cecropiifolia) fruit extract
Claudia Cordero1, Juliana Barrios2, Fabio Aristizabal1, Coralia Osorio*2, Alicia Lucia Morales2
1Departamento de Farmacia, Universidad Nacional de Colombia, Carrera 30 No 45-03, Edificio 450; Oficina 206. Ciudad Universitaria, Bogota DC
2Departamento de Quimica, Universidad Nacional de Colombia, AA 14490, Bogota DC, Colombia, cosorior@unal.edu.co

The methanolic extract of Pourouma cecropiifolia fruit, a tropical plant native from Amazon region, showed selective cytotoxicity in human carcinoma cell lines when evaluated by MTT assay. This extract was fractionated by using column chromatography on Sephadex LH-20 with a discontinuous gradient of methanol-water and acetone-water. As a result, three fractions were obtained, F1 to F3, which composition were determined by HPLC-PDA and LC-ESI/MS analyses. F1 was constituted mainly by anthocyanins, F2 presented a high amount of flavonols, and F3 was a polymeric fraction. F3 presented significant cytotoxic activity on HEp-2 (laringe carcinoma) and MKN-45 (gastric carcinoma) human carcinoma cell lines at the microgram per milliliter range, indicating a potential anticancer activity of this fraction.

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Evaluation of the total phenolic content in Brazilian peaches
Elisia R. Correa1, Maria do Carmo B. Raseira2, Jose Angelo Zuanazzi3, and David H. Byrne*4
1FAEM/UFPel, CAPES, Pelotas, RS, Brazil; 2EMBRAPA Clima Temperado, Br 392/Km 78. Pelotas, RS, Brazil; 3College of Pharmacy, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil; 4Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, Texas A&M University, College Station, TX, USA, d-byrne@tamu.edu

Fruits and vegetables are known to play an important role in human health. Sixty four peach [Prunus persica (Batsch) L.] genotypes (12 cultivars, 6 selections and 46 seedlings) originating from the EMBRAPA Clima Temperado Peach Breeding Program were characterized for their total phenolics. Five ripe fruit from each genotype were picked and liquefied and two and a half grams of this product (flesh and skin) were homogenized with 20 mL of methanol using a vortex mixer. Tubes were capped and stored overnight at 5C and then centrifuged for 30 min at 2000 rpm. A 50 uL aliquot of each prepared sample was diluted in 100 uL of nanopure water. A blank (control) containing 50 uL of methanol was equally diluted and analyzed. Each sample and blank were combined with 50 uL of 0.25 N Folin-Ciocalteau reagent, and allowed to react for 3 min before the addition of 100 uL 25 N Na2CO3. The reaction mixture was incubated for 2 h at room temperature and meas urements of absorbance at 725 nm were taken. The concentration of total phenolics was estimated from a chlorogenic acid standard curve and expressed as chlorogenic acid. Total phenolic content varied from 125 to 690 mg chlorogenic acid equivalents/100 g tissue (x = 427). The highest values were obtained in the fruit of the genotypes C-02-224-17 [(Bolinha x Cerrito)F2], Conserva 919 and Conserva 688 with 690, 680 and 671 mg chlorogenic acid equivalents/100 g tissue respectively.These levels of phenolics are higher than previously reported among yellow and white fleshed peaches and nectarines. Correlation analysis indicated that FDP (fruit development period), and flesh color (Hue angle) were not correlated and the ripening time was weakly correlated (r = 0.53) with total phenolics. The broad sense heritability for total phenolics content in this population of peaches was 0.46.

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Comparison of antioxidant activity of two soft fruit sources with different anthocyanin compositions
Elham Hosseini Beheshti*1, Steven Lund2, David D. Kitts1
1Food Chemistry and Toxicology Laboratory, Food Nutrition and Health, 2Wine Research Centre, Food Nutrition and Health 2205 East Mall, University of British Columbia, Vancouver, BC, V6T 1Z4, Canada, elhamh@interchange.ubc.ca

During the past decade, considerable scientific interest has been given towards understanding the health benefits of anthocyanins in different fruits and vegetables. Anthocyanins are polyphenolic pigments that belong to the flavonoid family and in general are water-soluble, nontoxic pigments that have been previously demonstrated to possess numerous bioactive properties in vivo and vitro systems. Former studies have reported many protective effects of anthocyanin on human health preservation, such as anti-inflammatory, anti-mutagenic and anti-genotoxic properties, improvement of night visual acuity and overall reduced risk to heart disease due to its antioxidant activity. Blackcurrant represents one of the most important and richest sources of phenolics. In particular, blackcurrant has been reported to contain four predominant anthocyanins which include cyanidin 3-glucoside, delphinidin 3-glucoside, cyanidin 3-rutinoside, delphinidin 3-rutinoside. In contrast, malvidin 3-glucoside, delphindin 3-glucoside, cyanidin 3-glucoside, petunidin 3-glucoside and peonidin 3-glucoside are the major anthocyanins found in grape. Although the antioxidant capacity of anthocyanins has been well established, less is known about the extent to which specific anthocyanin composition has on total antioxidant activity. The aim of the present study was to compare the antioxidant activity of two different soft fruits, blackcurrant and grape, which have distinctly different anthocyanin profiles. In addition we tested whether the antioxidant activity of crude fractions represents either the sum of total anthocyanin content or alternatively a synergy between different anthocyanins.
The total antioxidant capacity was measured by the oxygen radical absorbance capacity (ORAC) and ABTS assays. The anthocyanin profile of grape and blackcurrant were characterized by HPLC/MS coupled with a diode array detector. The concentration of individual anthocyanins in all of the berries was quantified with HPLC using standards. Our data showed that blackcurrant has higher (P<0.05) total anthocyanin concentration [28.25-74.94 mg/g of dry weight sample] and total phenolic content [90-160 mg GA/g of dry weight sample], compared with grape. Our result are consistent with the greater (P<0.05) antioxidant activity of blackcurrant compared to grape. We will also show both total anthocyanin content as well as the composition of individual anthocyanins in the fruits in order to make descriptive and quantifiable assessments of total antioxidant capacity of different berry sources.

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Inhibitory effect study of alpha amylase inhibitor from wheat (Triticum aestivum var. zarrin)
Masoud Haidari Zadeh
Iranian, m.haidarizadeh@uok.ac.ir

Plant alpha-amylase inhibitors show great potential as drug-design targets for treatment of diabetes and digestion disorderes.These inhibitors also known as sensitizing agents in human. The numerous form of alpha amylase inhibitors was reported. In this study alpha-amylase inhibitor was extracted from Iranian wheat cultivar (Triticum aestivum v zarrin), precipitated and purified by anion exchange fast protein liquid chromatography .Electrophoresis of purified protein showed 0.59 relative mobility .Total hydrolytic activity of human salivary and bacillus subtilis alpha amylase were inhibited 92.o1% and 78.99% respectively by collected purified alpha- amylase inhibitor.

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Antioxidant and antiradical activity of leaves of Polycarpaea corymbosa in vitro
Vijaykumar Ishwar Hukkeri*1, Mruthunjaya Kenganora2
1Herbals Research Division, Dr. Nargund Research Foundation, Dattatreya Nagar, BSK III Stage, 100 Feet Ring Road, Bangalore - 560 085, India.
2Department of Pharmacognosy and Phytochemistry, J.S.S College of Pharmacy, Mysore - 570 015, Karnataka, India, hukkeri_vi@rediffmail.com

The importance of reactive oxygen species and free radicals has attracted increasing attention over the past decade. Reactive oxygen species (ROS), superoxide (O2.) anion, hydrogen peroxide (H2O2), peroxyl (ROO-) radicals, and reactive hydroxyl (.OH) radicals, the nitrogen derived free radicals, nitric oxide (NO.) and peroxynitrite anion (ONOO.) are all continuously produced during normal physiologic events. However, they are removed by antioxidant defence mechanisms. There is a balance between the generation of ROS and inactivation of ROS by the antioxidant system in organisms. Under pathological conditions, ROS are over-produced and result in oxidative stress. These molecules are exacerbating factors in cellular injury and aging process and have been implicated in over a hundreds of diseased states which range from arthritis and connective tissue disorders to carcinogenesis, aging, infection and acquired immunodeficiency syndrome. In treatment of these diseases, antioxidant therapy has gained an immense importance. Antioxidants have been reported to prevent oxidative damage by free radical and ROS, and may prevent the occurrence of disease, cancer and aging. The medicinal properties of plants have been investigated in the recent scientific developments throughout the world, due to their potent antioxidant activities. Flavonoids and phenolic compounds widely distributed in plants have been reported to exert multiple biological effects, including antioxidant, free radical scavenging abilities, anti-inflammatory, anticarcinogenic, etc. Polycorpea corymbosa is a plant belonging to family Caryophyllaceae. The leaves of this plant are used in Indian traditional medicinal system as poultice, warm or cold over boils and inflammatory swellings and given with molasses in the form a pill in treatment of Jaundice. While general phytochemical screening, the plant showed good amount of phenolics. Since the plant is used in treatment of jaundice, as an astringent and in inflammatory swellings in traditional medicine and presence of phenolics in the plant inspired us to study its antioxidant properties. The antioxidant potency of hydro alcoholic extract of Polycarpaea corymbosa (PC) was investigated employing various established in vitro systems, such as DPPH radical scavenging, Nitric oxide (NO.) scavenging, β-Carotene linoleic acid module system (B-CLAMS), Hydroxyl (.OH.) radical scavenging, Anti lipid peroxidation. IC50 values were determined in each experiment. Further, total antioxidant capacity (TPC) and total phenolic content of PC was determined. IC50 values of PC were found to be 15.10 ± 0.10, 207.96 ± 32.77, 134.47 ± 21.39, 18.03 ± 3.22 and 46.73 ± 3.39 µg/ml in DPPH radical scavenging, NO. scavenging, B-CLAMS, .OH radical scavenging and Anti lipid peroxidation activity respectively. Phytochemical investigation revealed the presence of flavonoids. Acute toxicity study was also carried out to know whether it has toxicity. It was found that, Dose of 5000 mg/kg body wt. given for ten days the PC was found to be safe in mice and rats. The results indicate that PC possesses potent antioxidant activity and presences of flavonoids are found to be responsible for this activity.

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ISAFRUIT a large, multi-disciplinary European integrated fruit project 2006-2010
Lars Ove Dragsted*1, Niels Bent Bredmose2, and Ole Callesen2
1Department of Human Nutrition, Faculty of Life Sciences, University of Copenhagen, Denmark, ldra@life.ku.dk
2Department of Horticulture, Falculty of Agricultural Sciences, University of Aarhus, Denmark

ISAFRUIT is a trans-disciplinary, integrated European research project with the overall aim to provide knowledge that removes obstacles to fruit consumption. ISAFRUIT has a fork to farm, consumer driven design. The project is divided into eight research pillars, the first of which concerns consumer attitudes to fruit and fruit consumption where obstacles to consumer motivation or acceptance are identified. In other pillars, these identified obstacles as well as others identified in advance form the basis for research on health aspects, fruit processing, pre- and post harvest treatments and development of new varieties. Two extra pillars are concerned with dissemination of knowledge and project coordination. Overall coordinator of ISAFRUIT is Ole Callesen, Dept. of Horticulture, University of Aarhus. A total of 60 partners from Universities, independent research institutes, and private companies in 16 countries are engaged in ISAFRUIT research.
The project is basically concerned with all fruits but has focused mainly on stone fruits. Each pillar is subdivided into work packages (WPs) so that for instance in the first pillar research on fruit consumption trends and the forces that drive consumers to choose are subject of specific WPs. Others concern innovation acceptance among consumers and optimization of the supply chain. In pillar 2 on fruit and health our lacking understanding of why fruit is beneficial to health is the theme of one WP, specifically to understand better whether processed fruit also has the beneficial health effects of the fresh fruit. Other WPs concern a) fruit and obesity, b) low-allergenic fruit, and c) minimizing pesticide as well as fungal contaminants in the new ISAFRUIT pre- and post harvest treatments and innovated supply chain. Pillar 3 is concerned with optimization of fruit processing in order to improve processed fruit quality and to bring forward innovative processed fruit products in order to provide high convenience and maximal freshness of the new fruit products. Pillar 4 aims to identify better post harvest methods. One WP aims to provide sustainable non-chemical alternatives for pesticide use to control fruit losses by rot, another aims to develop and test a prototype of a decision support system (DSS) and a prototype of a portable, non-destructive multi-sensor device for fruit quality assessment. Pillar 5 aims to improve pre harvest methodologies, including minimal use of pesticides, better handling of other production resources and overall better orchard management to improve basic fruit quality. Pillar 6 is concerned with the development of novel fruit varieties, especially with an aim to improve the eating quality of the new varieties at the consumer end of the supply chain. Pillar 7 deals with several aspects of learning and dissemination, including maintenance of the ISAFRUIT website (www.isafruit.com) and courses to spread the knowledge from ISAFRUIT research to those responsible for the various parts of the fruit supply chains.

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Impact of deficit irrigation and plant population on yield, leaf quality and carotenoid content of spinach
Daniel I. Leskovar*, Shinsuke Agehara, Giovanni Piccinni, and Kil Sun Yoo
Vegetable and Fruit Improvement Center, Texas A&M University, Texas Agricultural Experiment Station, Uvalde, TX, USA, d-leskovar@tamu.edu

Field studies were conducted to determine yield, leaf quality and carotenoid content in response to deficit irrigation and plant population of spinach (Spinacia oleracea L. cv. DMC 66-16). Spinach seeds were planted during fall 2005 at various plant populations, ranging from 497,000 to 1,129,000 seeds/ha. Three irrigation regimes were imposed with either a Center pivot or low pressure drip system (LPS), 100%, 75%, and 50% crop evapotranspiration rates (ETc). Harvests were done on 28 February 2006. Total rainfall and irrigation received were 213, 170, and 132 mm in the Center pivot system, and 295, 244, and 198 mm in the the LPS for the 100%, 75% and 50% ETc, respectively. Under both systems there were significant differences for yield and quality components. Conversely, plant population and the interaction of irrigation rate and plant population were non significant for most measured parameters. In the LPS, marketable yield was not reduced by deficit irrigation at 75% ETc, but water use efficiency was significantly higher at 50% ETc compared to 100% ETc. In the Center pivot system, marketable yield was not reduced by deficit irrigation at 75% ETc, but was significantly lower at 50% ETc. In this system, water use efficiency was significantly higher at 75% ETc compared to 50% ETc. Petiole length, a quality component for the processing canning industry, was significantly reduced at 50% ETc for both systems. Ascorbic acid, beta carotene, lutein, and neoxanthin, consistently increased at 50% compared to 100% ETc rate under both irrigation systems, while violaxanthin content was unaffected by the irrigation rate treatment. The content of phytochemicals appeared to be enhanced under the Center pivot as compared to the LPS system, probably as a response to mild stress since the Center pivot system received an average of 43% less water (rainfall+irrigation) and had lower irrigation frequency than in the LPS.

148-P
Identification and confirmation of molecular markers and orange flesh color associated with major QTL for high beta-carotene content in muskmelon
A.B. Napier, S.O. Park*, H.Y. Hwang, and K.M. Crosby
Vegetable and Fruit Improvement Center, Texas A&M University, College Station, TX and Texas Agricultural Experiment Station, Texas A&M Univ., Weslaco, TX, USA, so-park@tamu.edu

Beta-carotene, also known as pro-vitamin A, is an essential nutrient for human health. Our objectives were to identify AFLP, RAPD, and orange flesh color (WF) markers associated with major QTL for high beta-carotene content using bulked segregant analysis in an F2 population derived from the muskmelon (Cucumis melo L.) cross of ‘Sunrise’ (white flesh color & low beta-carotene) x ‘TAM Uvalde’ (orange flesh color & high beta-carotene) in a greenhouse experiment, and confirm the associations of molecular and flesh color markers with QTL for this antioxidant trait in an F2 population from the same cross in a field experiment. Continuous distributions for beta-carotene were observed in the greenhouse and field F2 populations indicating quantitative inheritance for the trait. A significant positive correlation was found between beta-carotene and total soluble solids. Two DNA bulks were prepared from eight high and eight low beta-carotene greenhouse F2 plant, respectively. The bulks were screened for detecting polymorphic AFLP markers using 128 primer combinations. However, this AFLP technique did not provide useful polymorphisms. These bulks were also screened for identifying RAPD markers using 510 random primers. Seven RAPD markers and orange flesh color were detected to be significantly associated with QTL for high beta-carotene content in the F2 population of the ‘Sunrise’ x ‘TAM Uvalde’ cross in the greenhouse based on simple linear regression. Two unlinked markers associated with QTL were significant in a stepwise multiple regression analysis where the full model explained 59% of the total phenotypic variation for beta-carotene. Three RAPD markers and WF were confirmed in the F2 population of the same cross in the field to be consistently associated with QTL for the pro-vitamin A trait. These RAPD and flesh color markers associated with the beta-carotene QTL, consistently expressed in our populations under greenhouse and field environments, could be at least partially utilized in improving the level of this nutrient in new melon cultivars.

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Nutritional value, sensory qualities, and drug-interactive compounds of organically vs. conventionally grown ‘Rio Red’ grapefruit juice
Gene E. Lester
U. S. Department of Agriculture, Agricultural Research Service, Weslaco, TX, USA, gene.lester@ars.usda.gov

Most claims that organic produce is better tasting and more nutritious than nonorganic (conventional) produce are largely unsubstantiated. This is due mainly to a lack of rigor in research studies matching common production variables of both production systems; such as microclimate, soil type, fertilizer elemental concentration, previous crop, irrigation source and application, plant age, and cultivar. The aforementioned production variables common to both production systems were matched for comparison of Texas commercially-grown conventional and certified-organic ‘Rio Red’ red-fruited grapefruit. This rigorous matching of common production variables provided for a unique comparison of inputs specific to conventional vs. organic production systems affecting juice quality, including the comparative levels of human health-bioactive compounds, and consumer acceptance. Whole grapefruit from each production system were harvested between 0800 and 1000 hr at commercial early- (Nov.), mid- (Jan.) and late-season (March) harvest periods for three consecutive years. Within each harvest season conventional and organic juice was compared for nutritional qualities (ascorbic acid, lycopene, sugars and potassium), sensory qualities (tartness, sweetness, and overall acceptability), and human drug-interactive compounds (bergamottin, 6,7- dihydroxy bergamottin, 6,7-dihydroxy bergamottin dimmer 708, 6,7-dihydroxy bergamottin dimmer 728, naringin, narirutin, neohesperidin, and nitrate). Findings from this study have demonstrated production inputs specific to conventional vs. organic systems can significantly impact ‘Rio Red’ grapefruit juice nutritional qualities, consumer taste and acceptance, and human drug-interactive compounds. Conventional fruit was better colored, higher in lycopene, and the juice was less tart, lower in the bitter principle naringin, and better accepted by the consumer panel than organic. Whereas, organic juice was higher in ascorbic acid and sugars, and lower in the drug interactive bergamottin compounds and the negative health factor nitrate.

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Degradation of lycopene and β-carotene in model systems and in lyophilized guava during storage: Kinetics, structure and matrix effects
Jose Emilson M. Ferreira and Delia B. Rodriguez-Amaya*
Departamento de Ciência de Alimentos, Faculdade de Engenharia de Alimentos, Universidade Estadual de Campinas,
C.P. 6121, 13083-862 Campinas, SP, Brasil, delia@fea.unicamp.br

Being highly unsaturated molecules, carotenoids are highly susceptible to isomerization and oxidative degradation during the processing and storage of food. In the present study, the degradation of acyclic lycopene and dicyclic β-carotene in low-moisture model systems (carotenoids impregnated in potato starch, corn starch, or microcrystalline cellulose), simulating dehydrated foods, and in an aqueous model system (carotenoids dispersed in water), simulating juices, as well as in lyophilized guava, during storage at ambient temperature, in the presence (328 lux) or absence of light, was investigated. Both carotenoids followed first order kinetics under all the conditions studied. Light consistently and strongly promoted degradation under all conditions studied. Almost complete loss of lycopene (95-98%) occurred in the low-moisture model systems after 10 days of storage under light exposure, regardless of the solid support used. In the dark, there was 54 to 86% loss during the same period, the loss being greater in the corn starch system and lower in the potato starch system. In the aqueous system, lycopene loss was also greater under lighted storage (89%), but the difference with that of dark storage was small (81%). β-Carotene was much more stable than lycopene. β-Carotene losses of 71 to 92% occurred after 20 days under light, the loss being greater in microcrystalline cellulose and lower in corn starch. In the dark, losses ranged from 24 to 74% during the same period, the loss being also greater in microcrystalline cellulose with similar low losses in the corn and potato starches. Stability of lycopene isolated from guava, tomato and watermelon was compared in the potato low-moisture model system; greater losses were observed with lycopene from tomato. Since the model system was identical in the three cases, these results indicated that other compounds from the food sources, co-extracted with lycopene, might have influenced the oxidation. The considerable susceptibility of lycopene to degradation in model systems was not observed in lyophilized guava, demonstrating the substantial protective effect of the matrix. After 20 days of storage, double the storage time that caused almost complete destruction of this carotenoid in the low-moisture systems exposed to light, the loss was 77%. In the dark 65% lycopene was lost during the same period. The percent loss of β-carotene, found in guava at a concentration 18 times lower than lycopene, was similar, indicating that the initial concentration is another influencing factor.

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Impact of low water use systems on bioactive flavonoids of grapefruit
Ram M. Uckoo*1,2, Shad D. Nelson1,2, G. K. Jayaprakasha1 and Bhimanagouda S. Patil1
1Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, Texas A&M University-College Station, TX, USA
2Dept. of Agronomy & Resource Sciences, Texas A&M University-Kingsville, Kingsville, TX, USA, rammohanuckoo@yahoo.com

Citrus is grown in approximately 27,000 acres in Texas with Rio Red grapefruit comprising of almost 70% of the total acreage. The majority of citrus growers in Texas flood irrigate their orchards but due to rapid urban development, semi arid conditions and declining irrigation water supplies, alternative water conservative irrigation practices are necessary. In our recent field studies on citrus production under low water use systems, drip and micro-jet spray resulted in enhanced yields over flood irrigation. However, the impact of low water use systems on citrus bioactive flavonoids has not been reported. The objective of this study was to understand effect of irrigation practices on grapefruit bioactive flavonoids. Four flavonoids, naringin (NR), narirutin (NT), didymin (DY) and poncirin (PN) were quantified using HPLC from grapefruit grown under three irrigation systems: drip, micro-jet and flood irrigation. While fruit from drip irrigated trees had higher flavonoid content, no significant effect was noticed among low water use systems and flood irrigated trees. The ranges of NR, NT, DY and PN were 92-96, 28-31, 8.2-9.5 and 17-19 μg 100 mL-1 of fresh juice respectively. Total phenolic content of juice were determined spectrophotometrically using Folin-Ciocalteau reagent, and expressed as gallic acid equivalents. Total phenolic contents ranged from 4.8 to 5.5 μg g-1 of gallic acid equivalent with no significant effect due to irrigation treatments. The study demonstrates that low water use systems apart from enhanced yields and water conservation, also maintain the bioactive compounds of grapefruits.

152-P
Effect of worm castings, cow manure, and forest waste on yield and fruit quality of organic blueberry raised on a heavy soil
Girish K. Panicker*1, James M. Spiers2, Cedric A. Sims1, Juan L. Silva3, and Frank B. Matta4
1Department of Agriculture, Alcorn State University, Alcorn State, Mississippi, USA; 2USDA/ARS, Southern Horticulture Research Center, Poplarville, Mississippi, USA; 3Department of Food Technology, Mississippi State University, Mississippi State, Mississippi, USA; and 4Department of Plant and Soil Science, Mississippi State University, Mississippi State, Mississippi, USA, panicker@alcorn.edu

Blueberries are one of the richest sources of antioxidant phytonutrients of the fresh fruits and vegetables. Compared to other fruits, blueberries have a high level of anthocyanins. Anthocyanins are phenolic compounds that possess antioxidant activities. Rabbiteye blueberry (Vaccinium ashei Reade var. Tifblue) was grown on Memphis Silt Loam soil (Typic Hapludalf, silty, mixed, thermic). Two treatments of organic manures (worm castings W, cow manure C) were applied in basins around each plant. Control treatment received regular inorganic fertilizer. All treatments received pine bark and pine needle uniformly. No chemicals were applied to control pests, diseases, and weeds. Leaf area index (LAI), percent canopy cover, canopy width and height, stem diameter, and yield were significantly higher in organic plants treated with worm castings. There was no significant difference in size, diameter, degree brix, and vitamin C of the fruit, but the content of anthocyanins was higher in fruit treated with worm castings. No pathogenic organism from organic manures was found in fruits. Concentrations of nitrate-N and P were higher in surface soils treated with organic manures, but there was no trend in N or P enrichment in lower layers of the soil. The leaching of N and P into subsurface layers from inorganic fertilizer was highly significant. The results show that blueberry can be grown successfully on heavy soils with forest waste that can increase soil acidity and worm castings are excellent organic manure for this crop.

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Antioxidant activities in Kagzi lime (Citrus aurantifolia Swingle)
Jaiprakash R. Patil*1, G. K. Jayaprakasha1, Mahadev B. Chetti2, and Bhimanagouda S. Patil1
1Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, Texas A&M University, College Station, TX, USA, jrpatil_uas@yahoo.com
2University of Agricultural Sciences, Krishinagar, Dharwad, 580005, India

Limes have different local names in different countries such as Kagzi lime in India and Mexican lime or Key limes in United States. Limes contain several putative bioactive compounds. Some of these compounds are known to posesses antioxidant activity. In vitro studies associated with citrus have shown many biological activities. Our study focused on, antioxidant potentials of different lime extracts. The juice concentrate, seeds and peel powder was extracted with different solvents such as chloroform, ethyl acetate (EtOAc), methanol (MeOH), and MeOH:water. The extracts were concentrated, freeze dried and screened for their radical scavenging activity using DPPH and ABTS methods. The chloroform extract of lime juice concentrate showed maximum radical scavenging activity in both DPPH and ABTS assay at four concentrations. However, MeOH extract of seeds showed the highest radical scavenging activity. In contrast, EtOAc extract of lime peel exhibited maximum radical scavenging activity. The phenolic content was found to be highest in extracts of MeOH, EtOAc and MeOH: water, in juice concentrate, seeds and peel respectively as compared to other extracts. The above results suggest that limes posses health maintaining properties which can be explored further for potential disease prevention. This project is funded by the USDA-CSREES # 2006-34402-17121 “Designing Foods for Health” through the Vegetable & Fruit Improvement Center.

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Protection rendered to primary rat hepatocytes against oxidative stress by anthocyanins from Syzygium cumini fruit
Jyothi Maria Veigas*, Mandayam S. Narayan and Bhagyalakshmi Neelwarne
Plant Cell Biotechnology Department, Central Food Technological Research Institute, Mysore-570 020, India, veigasmaria@yahoo.com

Syzygium cumini Skeels (Myrtaceae) is a tropical tree producing edible berries commonly known as Indian black plum (syn. Java plum). The berries are rich in anthocyanin pigments that are concentrated in the peel rather than in the sweet pulp. This study was designed to understand the cytoprotective activity of the anthocyanin rich extract from fruit peels of S. cumini. The total anthocyanin content being 230mg/100g (DW) contained largely glucoglucosides of delphinidin, petunidin and malvidin that were identified by HPLC-ESI-MS. Screening for bioactivities using chemical in vitro models revealed strong antioxidative potential of the peel extract. To ascertain the mechanisms through which the anthocyanins impart antioxidative effects, cultured cell model of primary rat hepatocytes challenged with tertiary butyl hydroperoxide (TBH) - a free radical generator- was used. Cellular changes were estimated using biochemical indices such as lactate dehydrogenase leakage, lipid peroxidation, reduced glutathione content (GSH), activities of endogenous antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) and DNA damage. The extract was found to be non-toxic to cultured rat hepatocytes and it enhanced the viability of the TBH-treated cells by up to 65% at the tested concentrations (50-500 ppm). LDH leakage was suppressed to an extent of 50% at 250 ppm, which is indicative of the protection rendered against TBH-induced oxidative insult. TBH significantly reduced the activity of GPx whereas CAT and SOD were not substantially affected. However, treatment with the S. cumini peel extract (100 and 250 ppm) markedly elevated the enzyme activities in TBH-treated cells. These observations suggest that the anthocyanins of S. cumini fruit peel exert a protective effect against TBH-induced oxidative stress in cultured primary rat hepatocytes, not just by scavenging the free radicals generated (as observed in our previous study), but also by modulating the antioxidant enzyme activities. Further studies done on the effect of extract on the expression of mRNAs of the antioxidant enzymes are discussed.

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Suppression of Escherichia coli O157:H7 type III secretion system by citrus limonoids
Amit Vikram*1, Palmy R. Jesudhasan2, Suresh D. Pillai1, 2, G. K. Jayaprakasha1, Bhimanagouda S. Patil1
1Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, 2Department of Poultry Science, Texas A&M University, College Station, TX, USA, AVikram@ag.tamu.edu

Citrus limonoids are complex triterpenoids which have displayed significant biological activities. In current investigation, the suppression potential of citrus limonoids on Escherichia coli O157:H7 virulence was explored. Cell-to-cell signaling is known to regulate virulence and shiga toxin production in E. coli O157:H7, an important intestinal pathogen of humans, which causes histopathological lesions on intestinal epithelial cells. Shiga toxin is responsible for major symptoms of hemorrhagic colitis and hemolytic uremic syndrome. Citrus limonoids such as isolimonic acid, ichangin, isoobacunoic acid, isoobacunoic acid glucoside and deacetyl nomilinic acid glucoside were purified from sour orange seeds using open column chromatography. Expression of 9 genes, encoded within Locus of Enterocyte Effacement (LEE), shiga toxin and flagellar synthesis were measured upon exposure to five limonoids. E. coli O157:H7 was grown until OD600 1.2, total RNA was extracted and reverse transcribed to obtain first strand cDNA. Expression of genes was studied using quantitative real-time PCR. Fold change was calculated using 2ΔΔCT method. Isolimonic acid and ichangin strongly attenuated the expression of LEE encoded genes responsible for type III secretion, as well as shiga toxin stx2 gene and flagellar genes flhDC. FlhDC are transcriptional regulators of flagellar biosynthesis and loss in activity of these two genes has been implicated in the inhibition of motility and colonizing ability of bacteria. Stx2 was down regulated 4.7 and 2.4 fold by isolimonic acid and ichangin respectively. These results indicate that application of isolimonic acid and ichangin may reduce the efficacy of E. coli O157:H7 to infect the host. While isoobacunoic acid, isoobacunoic acid glucoside and deacetyl nomilinic acid glucoside down regulated the LEE region genes, their potency was less compared to isolimonic acid and ichangin. Our results suggest that citrus limonoids are potent antagonist of bacterial cell-to-cell signaling. Furthermore, limonoids may help in prevention of bacterial infection. This project is based upon work supported by the USDA-CSREES # 2006-34402-17121 “Designing Foods for Health” through the Vegetable & Fruit Improvement Center.

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Radish leaves and radish roots: are we eating the right part for cancer prevention?
1Tim O’Hare*, 1Lung Wong , 2Lesleigh Force
1Queensland Department of Primary Industries and Fisheries; 2University of Queensland, oharet@dpi.qld.gov.au

Radishes are most commonly consumed as a root vegetable, although radish leaves are occasionally used in salads and cooking. While both the radish root and shoot contain glucosinolates with anti-cancer potential, the glucosinolate profile of the root and the shoot are very different. Whereas the root contains mainly glucodehydroerucin (2.8 μmol/gFW) (also known as glucoraphasatin), the main glucosinolate components of the shoot are glucoraphanin (3.3 μmol/gFW) and glucoraphenin (1.7 μmol/gFW). Upon hydrolysis, the latter glucosinolates produce sulforaphane and sulforaphene respectively, both potent inducers of mammalian phase 2 enzymes. Previously, radishes have been dismissed as having minimal anti-cancer potential, based on studies with radish roots. However, depending on cultivar, radish leaves can have up to 40 times the capacity of roots to induce phase 2 enzymes. In fact, radish leaves have similar anti-cancer potential to mature broccoli, a vegetable that has received considerable interest in this area. Despite this, the acceptance of radish leaves is likely to remain limited due to culinary factors such as flavour and texture.

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Determination of chemoprotective D-glucaric acid levels in grapefruit (Citrus paradise Macf.) varieties using HPLC
Jose Luis Perez*, G.K. Jayaprakasha, and Bhimanagouda S. Patil, Vegetable & Fruit Improvement Center, Department of Horticulture Sciences Texas A&M University, ppmd04@gmail.com

Grapefruit has potential health promoting properties due to the presence of multitude bioactive compounds. Ongoing cell culture and animal studies in our lab using limonoids and flavonoids have provided strong evidences of their protective properties of preventing chronic diseases. Studies related to D-glucaric acid, a natural, nontoxic bioactive compound found in grapefruit has not been explored. One of the derivatives, such as D-glucaro-1, 4-lactone, is reported to be a potent β-glucuronidase inhibitor. With the inhibition of β-glucuronidase enzyme, glucuronidation will be favored. Glucuronidation is a conjugation process through which potentially carcinogenic environmental toxins can be neutralized. In this context, method for quantification of D-glucaric acid using HPLC was developed. Nine grapefruit varieties were examined. Grapefruit samples were homogenized, centrifuged, and supernatant filtered. Filtrate was analyzed by HPLC using an isocratic mobile phase (0.1% phosphoric acid). Levels of D-glucaric acid were found to range from 25-126.8 mg/100ml of grapefruit homogenate. This project is based upon the work supported by the USDA CSREES IFAFS #2001-52102-02294 and USDA-CSREES # 2005-34402-14401 “Designing Foods for Health” through the Vegetable & Fruit Improvement Center.

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Late-season sour orange seeds are a major source of deacetylnomilin
Haejin Bae*1, G.K. Jayaprakasha1, John Jifon1,2 and Bhimanagouda S. Patil1
1Vegetable and Fruit Improvement Center, Department of Horticulture Sciences, Texas A&M Unviersity, College Station, TX, USA, 2Texas Agriculture Experiment Station, Weslaco, TX, USA, hjbae@tamu.edu

The genus Citrus contains several biologically-active limonoids that are unique distributed mainly in the Rutaceae family. Deacetylnomilin and other limonoids have been shown to possess various biological activities such as anti-proliferation of neuroblastoma cells, colon cancer and human breast cancer cell lines. Deacetylnomilin has also been shown to have antifungal and antibacterial properties. The distribution of deacetylnomilin varies greatly among various citrus species. In the present work, isolation of the limonoid aglycones from seeds of sour orange (Citrus aurantium L.), a widely-used rootstock species, was investigated. Seed powder (6.9 Kg) was extracted in a Soxhlet extractor with hexane, and ethyl acetate consecutively for 16h. The ethyl acetate extract was concentrated and loaded on silica gel column, and eluted with different mobile phases. All the fractions were analyzed for putative compounds of interest using TLC and HPLC. Fractions containing similar spots were pooled and crystallized. The total yield of deacetylnomilin was 4.9 g per 6.9 Kg dry wt. Purity of the isolated compound was assessed by HPLC using a C18 column and detection at 210 nm. Further, the structure of this compound was confirmed by mass spectrometry. This is the first report demonstrating significant yields of deacetylnomilin from late harvested sour orange seed. This study also reveals that significant quantities of deacetylnomilin can be isolated late season fruit, hence, seeds from such fruit may be a better source of deacetylnomilin for bioactivity assay studies. This project is based upon the work supported by the USDA CSREES IFAFS #2001-52102-002294 and USDA-CSREES # 2006-34402-17121 “Designing Foods for Health” Through the Vegetable & Fruit Improvement Center.

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Isolation of bioactive compounds from grape fruit juice concentrate
Kranthi Chebrolu*, G.K. Jayaprakasha, John L. Jifon1, and Bimanagouda S. Patil
Vegetable and Fruit Improvement Center, Department of Horticulture Sciences, 1Texas A&M Unviersity, College Station, TX, USA, Texas Agriculture Experiment Station, Weslaco, TX, USA, scorp_kranthi@yahoo.com

Citrus fruits are considered to have various health benefits including prevention of several types of cancers. These properties could be attributed to the antioxidant secondary metabolites present in the fruits such as bergaptol, β- sitosterol, β-sitosterol glucoside and limonin. Recent investigations using bergaptol has reported a very high radical scavenging ability for it in addition to being a potent CYP 3A4 enzyme inhibitor. In another study β-Sitosterol was shown to induce anti-proliferation and apoptosis of fibrosarcoma cells. Although the secondary metabolites are being utilized for several investigations, the isolation of pure compounds in sufficient quantities has been challenging. To this end, in the current study an attempt has been made to establish a procedure to separate several of the abovementioned bioactive compounds from grape fruit juice concentrate. Grape fruit juice concentrate was extracted using Methyl Ethyl Ketone (MEK), and the extract was further subjected to silica gel column chromatography using mixtures of organic solvents as mobile phases. TLC and HPLC techniques were used to identify the compounds in the fractions collected. Bergaptol (986mg), β-sitosterol (456mg), and limonin (49mg) were successfully isolated from the column fractions by crystallization techniques when using mixtures of hexane and chloroform at10:90, 25:75, 40:60 ratios respectively as the mobile phases. Furthermore β-sitosterol glucoside (384mg) was isolated using chloroform and acetone (90:10) mixture. In conclusion in the current study we report the isolation of several bioactive compounds in quantities sufficient to facilitate further investigation of their therapeutic abilities along with their purification procedures. Future experiments in isolating several other bioactive compounds may thus provide opportunities to elucidate the mode of action of these compounds in detail and may reinforce the potential health benefits of Citrus fruits.

160-P
Temporal, dose and interaction effects of methyl jasmonate, salicylic acid and resveratrol on total antioxidant capacity in table grape (Vitis vinifera L. cv Crimson Seedless) postharvest
Michael Considine*1, Colin Gordon1, Kevin Croft2 and Simon Ching3
1Department of Agriculture and Food Western Australia, Locked Bag 4, Bentley DC, Western Australia, 6983, Australia, 2School of Medicine and Pharmacology, University of Western Australia, Perth, Western Australia, 3PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Nedlands, Western Australia 6009, Australia , mconsidine@agric.wa.gov.au

The pervasive plant signalling molecules jasmonate and salicylate are well recognised for their involvement in plant pathogen and herbivore defence. The phytoalexin resveratrol and its derivatives have also been widely studied in relation to plant defence but have become increasingly renowned for their dietary health potential. Previous studies have documented the elicitation of various antioxidants by exogenous application of each of these molecules in plants. Yet, jasmonate and salicylate have been shown to be antagonistic in their signalling mode and effect. Here we have applied various concentrations of salicylic acid and methyl jasmonate, alone or in combination, as well as one concentration of trans-resveratrol, to harvested table grapes (Vitis vinifera L. cv Crimson Seedless) prior to cool storage. Whole berries were sampled in a time-course over eight weeks. We have investigated dose and temporal responses of the grapes by a novel total antioxidant assay, Antioxidant Inhibition of Oxygen Radicals (AIOR; Ching et al., 2006). We found that both salicylic acid and methyl jasmonate had significant dose-responses when applied alone, and had a significant interaction, resulting in a positive effect on AIOR. At the time of submission, further data was being analysed. The results are discussed in terms of the potential to manipulate the total antioxidant capacity of grapes through the use of these elicitors, alone or in combination.

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Colour retention and texture in fresh-cut apple slices held at 2-4oC for five days
Derek Keenan*1, Aidan O Duinn1, Ronan Gormley1, Francis Butler2
1Ashtown Food Research Centre, (Teagasc), Ashtown, Dublin 15, Ireland; 2Biosystems Engineering, School of Agriculture, Food Science and Veterinary Medicine, University College Dublin, Dublin 2, Ireland, derek.keenan@teagasc.ie

Fresh fruit salads are an increasing feature in retail outlets due to consumer demands for convenience foods. The current study addressed the issue of colour retention, with and without the use of a browning inhibitor, in fresh cut slices for 12 apple cultivars, i.e. Granny Smith, Golden Delicious, Bramley’s Seedling, Braeburn, Jonica, Jonagold, Alwa, Cortland, Elstar, Shampion, Gloster, and Rajka. Slice texture was also measured as were soluble solids (%SS), titratable acidity (TA), and pH. Slices (6 mm thick) were dipped in a solution (6% w/w; 2 min) of NatureSeal AS1 (AgriCoat, UK) and were packed in clear trays (15 x 10.5 x 3 cm) with a heat sealed film (O2 transmission of <2ml /24hr /23°C and a water vapour transmission of <6 g/24 hr/38°C) followed by storage at 2-4°C for 5 days. A water dip was used as a control. Slice surface colour was measured with a Hunter colour meter (Lab), and slice shear values with the Kramer shear press (100g samples; standard test cell). The colour difference values (∆E) between days 0 and 5 was calculated and were higher for all cultivars in the absence of the browning inhibitor. The highest ∆E values in the absence of inhibitor were for Jonica (11.7) and Cortland (11.4) and the lowest for Shampion (2.3) and Elstar (4.3). Values (∆E) with the inhibitor were highest for Alwa (7.5) and Cortland (3.6) and lowest for Jonagold (0.71) and Gloster (1.7). In the case of the shear values, the presence of the inhibitor gave firmer slices on both days 0 (2080 vs 1595 N) and 5 (2429 vs 1898 N), presumably due to cross linking caused by calcium in the inhibitor. Shear values for the cultivars ranged from 2879 (Gloster) and 2268 N (Shampion) to 1671 (Elstar) and 1827 N (Jonagold). Titratable acidity was highest for Jonagold (7.6) and Rajka (5.8) and lowest for Shampion (3.4) and Elstar (4.3 meq/10g). The use of the inhibitor raised SS levels (12.6 vs 11.7 %) and Jonagold (14.0) and Rajka (13.2%) had the highest values of the cultivars. These data shows that the inhibitor had a positive effect on colour retention in all the cultivars tested and also had a firming effect and as such is a valuable process aid for fresh-cut apple slices. These trials are continuing with emphasis on apples and other fruits as constituents of fresh-cut fruit salads. This work is a component of the ISAFRUIT Project [Contact No. 06279 (food)] which is part-funded by the EU 6th Framework Programme.

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Serum antioxidant carotenoids inversely associate with serum liver enzymes: cross-sectional study from the Mikkabi study
Minoru Sugiura*1, Mieko Nakamura2, Hikaru Matsumoto1, Yoshinori Ikoma1, Kazunori Ogawa1, Masamichi Yano1
1National Institute of Fruit Tree Science, Shizuoka, Japan, 2National Institute for Longevity Sciences, Aichi, Japan, msugiura@affrc.go.jp

Introduction: Antioxidant micronutrients, such as vitamins and carotenoids, exist in abundance in fruits and vegetables and have been known to contribute to the body’s defense against reactive oxygen species. Numerous epidemiologic studies have demonstrated that a high dietary consumption of fruits and vegetables results in lower risks of certain cancers and cardiovascular disease. Liver plays an important role in metabolism and has a number of functions in the body including glycogen storage, plasma protein synthesis, and drug detoxification. Recently, it became known that antioxidant vitamins and carotenoids are reduced in several liver diseases, such as alcohol-induced hepatitis and cirrhosis. It is well known that alcohol induces the generation of free radical species during its metabolism. On the other hand, hyperglycemia such as diabetes increases the generation of free radicals by glucose auto-oxidation, and the increment of free radicals may also lead to liver cell damage. Oxidative stress is thought to play a key role in the pathogenesis of liver injury. Therefore, antioxidant micronutrients would be expected to protect against liver injury. In this study, we examined the associations of serum antioxidant carotenoids and serum liver enzymes stratified by alcohol intake levels or glucose tolerance status.
Methods: Data used in the present study were derived from health examinations of residents of the town of Mikkabi, Shizuoka Prefecture, Japan, in April and May 2003. Participants were recruited for this study, and informed consent was obtained from 886 subjects (302 male and 584 female). This study was approved by the ethics committee of the National Institute of Fruit Tree Science. The participants were divided into three groups according to their alcohol intake level or into two groups according to their glucose tolerance status. The associations of serum gamma-glutamyltransferase (gamma-GTP) or aspartate aminotransferase (AST) and alanine aminotransferase (ALT) with serum-carotenoid concentrations were evaluated cross-sectionally stratified by alcohol intake levels or glucose tolerance status, respectively. The multivariate-adjusted geometric means of the serum liver enzymes in each tertile of the serum-carotenoid concentrations were calculated after adjustment for confounding factors. Results: The serum gamma-GTP concentrations were significantly high in accordance with the ethanol intake level. In moderate and heavy drinkers (>=25 g/day), the multivariate-adjusted geometric means of serum gamma-GTP concentrations were significantly low in accordance with the tertiles of the serum lycopene, alpha-carotene, beta-carotene, and beta-cryptoxanthin concentrations. On the other hand, the serum AST and ALT concentrations in the groups of impaired fasting glucose (IFG) and diabetes were significantly higher than those in the normal group. The multivariate-adjusted geometric means of the serum AST and ALT concentrations in IFG and diabetes group were significantly low in accordance with the tertiles of the serum beta-cryptoxanthin and beta-carotene concentrations. The most inverse association of serum liver enzyme and carotenoid concentration was observed in beta-cryptoxanthin. Conclusions: The serum antioxidant carotenoids were inversely associated with alcohol- and hyperglycemia-induced increases of serum liver enzymes. Our results provided support for the hypothesis that antioxidant carotenoids may protect against oxidative stress induced by alcohol or hyperglycemia in the earlier pathogenesis of liver dysfunction.

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Formulating functional popsicle with wine and grape juice by-products
Emília Y. Ishimoto1, Vanessa Capriles1, Andréa G. Matias1, Aldo Baccarin2, Elizabeth A.F.S. Torres*1
1School of Public Health, University of São Paulo, São Paulo, Brazil; 2Food Intelligence-Consultoria Técnica em Alimentos S/C Ltda, São Paulo, Brazil, eatorres@usp.br

Consumers nowadays have increased substantially their demands for foods with high nutritional value. Grape pomace (GP) is the most important by-product of wine and grape juice production, and is also a good source of bioactive compounds, especially phenolic antioxidants and fiber. Therefore, GP can be considered a cheap source of functional ingredients, and its use can also reduce the environmental impact. Objective: To produce popsicle with wine and grape juice by-products and to evaluate its nutritional value and sensory acceptability. Materials and methods: Red GP (skin and seeds) were supplied by a winery and a fruit juice company, providing Cabernet Sauvignon and Izabel grape varieties, respectively. To obtain the pomace flour, the samples were freeze-dried, milled and sifted. The popsicles were elaborated in a pilot plant. Grape pomace juice popsicle (GPJP) and grape pomace wine popsicle (GPWP) were evaluated in color, texture, flavor and overall acceptability by forty five consumers in a 1 to 9 hedonic scale. Color was evaluated by the CIE L*a*b* system and the total color difference between the traditional and the developed popsicles was calculated as follow: Δ E = [ΔL*2 + Δa*2 + Δb*2]1/2. Nutritional value was estimated and also compared to a market available popsicle made with grape juice. Results: GPJP presented lower total color difference (ΔE=8.50) than GPWP (ΔE=12.76), which was responsible for its higher color acceptability (7.81±1.19), when compared to GPWP (5.98 ±2.30). Also, texture, flavor and overall acceptability of GPJP (6.81±1.47, 6.96±1.52, 6.94±1.31) were significant higher than GPWP (5.02±2.27, 5.51±2.09, 5.28±2.03). GPJP was significantly better preferred (p<0.001) regardind the sensory attributes. Total dietary fiber contents of the developed products is approximately 2,0 g/portion (= 1 popscicle or 60 g), while a market commercial popsicle in Brazil presents no significant amount in this nutrient (the fiber contents is not declared on the label). Conclusion: In this first use of grape by-products in food product development, we observed a viability of using grape by-products as an ingredient to make high phenolics and fiber popsicles with high sensory acceptability. The results showed a great potential to commercialize these products, especially for people concerned about their dietary needs. Support: FAPESP and CNPq. Acknowledgements: Vinícola Miolo Ltda, Produtos Alimentícios Superbom Ind e Com Ltda, Food Intelligence Consultoria Técnica em Alimentos Ltda.

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L-Ascorbic acid (vitamin C) loss in rose hip (Rosa canina L.) fruit during pulp production
Mustafa Karhan
Akdeniz University, Faculty of Agriculture, Department of Food Engineering, 07059 Antalya, Turkey, mkarhan@akdeniz.edu.tr

Ascorbic acid loss in rose hip (Rosa canina L.) fruit, which is one of the richest sources of ascorbic acid, was investigated during industrial pulp production. Rose hip, belonging to the Rosaceae family, is naturally grown in Central Asian and Anatolian territories. Due to the hairy structure covering the hard seeds, the fruit should be grinded, mashed and heated by hot water blanching in order to process into pulp. Ascorbic acid, known as vitamin C, is a temperature sensitive molecule. This water-soluble vitamin that serves to influence oxidation-reduction reactions is an important nutrient and essential to human being. The material used in this experiment was taken from a local factory processing rose hip fruit. The raw material was grinded by using a fruit grinder and immediately put into a mashing tank containing hot water at 85°C under anaerobic condition. After 1 hour, the mash was taken to pulper with 1.5, 1.2 and 0.5 mm pore size under steam media. Ascorbic acid contents of the samples taken from each process stage were then determined by RP-HPLC. Our results showed that the highest degradation of ascorbic acid occurred in the grinding stage.

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Effect of variety, cultivation and storage conditions on Mal d 1 protein of apple fruits
Anne Matthes*1, Michaela Schmitz-Eiberger2, Stephan Scheurer3, Anne-Regine Lorenz4, Gerhard Baab5, Ulrich Mayr6
1Bonn University, 2Bonn University, 3Paul-Ehrlich-Institute, 4Paul-Ehrlich-Institute, 5DLR Rheinpfalz, 6Kompetenzzentrum Obstbau-Bodensee, amatthes@uni-bonn.de

The prevalence of allergies is increasing throughout the world, this development can also be seen in a rising number of patients showing reactions to birch pollen related food allergens. Today up to 70% of birch-pollen-allergic patients may be food sensitive, most frequently to apples. The parallel appearance of birch and food allergy can be explained by cross-reactive IgE. Consumption of apples can provoke several allergic reactions due to the presence of the apple allergen Mal d 1. Allergic symptoms are limited to the oral allergy syndrome. The major birch pollen allergen Bet v 1 and the in Central Europe most important apple allergen Mal d 1 share allergenic epitopes leading to these IgE cross-reactivities. Mal d 1 has been identified as a 18 kD protein, present in pulp and peel of apple fruits. Both Bet v 1 and Mal d 1 belong to the so called pathogenesis-related (PR) 10 proteins, a family of proteins that are induced by pathogens, wounding, or certain environmental stresses. The biological role of Mal d 1 is still unknown, it may be involved into binding and transport of plant steroids. The allergenic composition of most apple cultivars has not yet been fully characterized. Data on the IgE binding potency of Mal d 1 in vitro and in vivo showed that this allergen is influenced by cultivar, degree of maturity and storage conditions of the fruits. But the amount of Mal d 1 depending on these factors and the effect on human health has not been determined so far. The aim of the study was to quantify Mal d 1 content of the fruits depending on cultivation method (organic and integrated production), cultivar and different terms of postharvest storage. Fruits of different apple cultivars (Malus domestica) were cultivated at the Research Stations Klein-Altendorf and Bavendorf, Germany. Optimal harvest date was determined by measuring values for firmness, starch degredation and sugar content (Streif-Index). Fruits were picked from definite positions of the tree for each variety and location. Protein extracts were prepared from the apple fruits after removing the core according to Bjoerksten et al. 1980. Mal d 1 content was quantified by a Sandwich-Enzyme-linked Immunosorbend Assay. Apple cultivars differ considerably in their Mal d 1 content. Highest amounts of Mal d 1 were determined in Golden Delicious and Topaz fruits. The lowest content was found in the cultivars Braeburn and Jonagold. At harvest the detected concentration of Mal d 1 was low, but during storage the Mal d 1 content increased significantly.
Bjoerksten F., Halmepuro L., Hannuksela M., Lahti A. (1980): Extraction and Properties of Apple Allergens. Allergy, 35, 671-677.

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Maternal knowledge, attitude and practice regarding iron intake during the periconceptional period
Kassim.H.M. Al-Awwad* and J.S. Dhahir
Dept. Food Science and Nutrition, College of Allied Sciences University of Applied Sciences 11931, Shafa Badran, Amman, Jordan, kassim_alawwad@asu.edu.jo

Three hundred and ninety two pregnant women from mother and child health care units and antenatal clinic were chosen for this questionnaire-based prospective study to evaluate the level of knowledge, attitude and practice regarding iron in nutrition and iron intake during the periconceptional period. The results showed that most pregnant women (346/392, 88.3%) had heard of iron in nutrition, women with higher education (p<0.01), those who had >3 children (p<0.01), higher income (p=0.05) and smoker (p=0.05) were more likely to be correctly informed. A minority (111/392, 28%) knew that iron could prevent anemia during pregnancy, those with higher education (p<0.001), their husband’s occupation related to nutrition or health (p<0.01), employed women (p<0.01), those who had >3 children (p=0.025), and with higher income (p<0.01), were more likely to be thus aware. One hundred thirty one (33.4%) knew the good food sources of iron, and was more likely in those with higher education (p<0.01), their husband’s occupation related to nutrition or health (p<0.01), those who had >3 children (p=0.05), with higher income (p<0.01), and with planned pregnancy (p<0.01). Minority of women (23/392, 5.9%) knew the right requirement of the pregnant women form iron, and was more likely in those with higher education (p=0.05), their husband’s occupation related to health or nutrition, (p=0.042), who had > 3 children (p=0.017), with higher income (p<0.01), and those with planned pregnancy (p<0.01). Minority of women (146/392) knew the right relationship between the iron and vitamin C and was more likely in women with higher education (p=0.042), their husband’s occupation related to health or nutrition(p=0.05), employed women (p=0.05), and with higher income (p=0.024).
Key Words: food conception, iron, maternal, health status

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Potential chemo-preventative properties isolated from sweetpotato (Ipomoea batatas L.) leaf
Shahidul Islam* and James O. Garner
Department of Agriculture, University of Arkansas at Pine Bluff, USA, islam@uapb.edu

Our previous experiment revealed that sweetpotato leaves were an excellent source of antioxidative polyphenolics compared to other commercial vegetables. The aim of the present study was to clarify the chemical characteristics of the polyphenols, and to clarify the relationship between the polyphenolic quality and quantity. Sweetpotato leaves contain at least fifteen biologically active anthocyanins that have significant medicinal value for certain human diseases and may also be used as natural food colorants. The anthocyanins were acylated cyanidin and peonidin type. But the content of cyanidin in leaves is much higher that that of peonidin, suggesting that the anthocyanin composition of sweetpotato leaf is cyanidin type. The cyanidin type anthocyanins are superior to the peonidin type in antimutagenicity and antioxidative activity. Structure-activity studies show that the number of sugar units and hydroxyl groups on aglycons is associated with biological activities of anthocyanins. The activities appear to increase with a decreasing number of sugar units, and with an increasing number of hydroxyl groups on aglycons. The six different polyphenolic compounds were identified and quantified by NMR, FAB-MS spectra and RP-HPLC analysis procedures. The result suggests that the main phenolic compound in Ipomoea batatas leaves is 3, 5-di-O-caffeoylquinic acid followed by 4, 5-di-O-caffeoylquinic acid. The results revealed that total polyphenol was positively correlated with the different caffeoylquinic acid derivatives of sweetpotato leaves. The result also suggests that all the six different bioactive caffeoylquinic acid derivatives and total polyphenol were positively correlated with the RSA of sweetpotato leaves as following order (n=60): total polyphenols (r=0.92; P<0.001) > 3,4-di-o-caffeoylquinic acid (r= 0.77; P<0.001) > 4,5- di-o-caffeoylquinic acid (r=0.74; P<0.001) > chlorogenic acid (r=0.74; P<0.001)> 3,5- di-o-caffeoylquinic acid (r=0.73; P<0.001)> 3,4,5-tri-o-caffeoylquinic acid (r=0.39; P<0.01) > caffeic acid (r=0.37; P<0.01). These caffeoylquinic acid derivatives (CQAD) showed very high 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity (RSA), and effectively inhibited the reverse mutation induced by Trp-1 on Salmonella typhimurium TA 98 and TA 100. The physiological function of CQAD with the plural caffeoyl group is more effective than with a monocaffeoyl one. The radical scavenging activity and the anti-mutagenicity of these derivatives in order of efficacy is triCQA > diCQAs > monoCQA, suggesting that the number of caffeoyl groups bound to QA plays a role in the radical scavenging activity of the CQA derivatives. In other words, additional caffeoyl groups bound to QA are necessary for higher function. Thus, the Ipomioea batatas leaves contained distinctive polyphenolic components with high content of mono-, di-, and tricaffeoylquinic acid derivatives and could be a source for bioactive compounds. The present experimental results showed that sweetpotato leaf contains a very high concentration of biologically active compounds, which may have the significant medicinal values for certain human diseases linked to oxidation, such as cancer, hepatotoxicity, allergies, aging, human immunodeficiency virus, and cardiovascular problems.

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Dietary limonin suppresses antigen-induced CD4+ T-cell proliferation via down-regulation of NF-kB nuclear translocation
Wooki Kim1,2, Roger Smith3, Lan Zhou4, Naisyin Wang4,5, G.K. Jayaprakasha, Bhimanagouda S. Patil6, David N. McMurray2,5,7, Robert S. Chapkin*1,2,5
1Department of Nutrition and Food Science, 2Intercollegiate Faculty of Nutrition, 3Department of Veterinary Pathobiology, 4Department of Statistics, 5Center for Environmental and Rural Health, 6Vegetable Fruit Improvement Center, Texas A&M University, 7Department of Microbial & Molecular Pathogenesis, Texas A&M University Health Science Center, College Station, TX, USA, r-chapkin@tamu.edu

We have previously demonstrated that dietary n-3 polyunsaturated fatty acids present in fish oil (FO) suppress inflammation in part by altering CD4+ T-cell membrane composition and co-localization of PKC-theta into lipid rafts (1). With respect to plant-derived bioactive food components, Limonin (a component of grapefruit) has been shown to down-regulate COX-2 and iNOS, which are target genes of nuclear factor (NF)-kB in colonic epithelial cells (2). In addition, Curcumin (diferuloylmethane), a yellow pigment extracted from the root of Curcuma longa L., can suppress inflammation in part by inhibiting NF-kB nuclear translocation (3). Therefore, we hypothesized that limonin and curcumin would enhance the anti-inflammatory properties of FO by suppressing NF-kB activation in T-cells. To determine the anti-inflammatory effect of dietary FO combined with either limonin or curcumin, DO11.10 mice, which express the transgenic T cell receptor (TcR) specific for OVA323-339 peptide, were fed a wash-out corn oil (CO, 5% CO) control diet for 1 wk, followed by either CO, FO (1% CO+4% FO), FO with 2% curcumin (Cur), or FO with 0.02% limonin (Lim) diets for 2 wks. Purified CD4+ T-cells were labeled with CFSE and stimulated with either (i) media control, (ii) mitogens (anti-CD3 and anti-CD28 mAbs), (iii) PMA and Ionomycin, or (iv) transgenic TcR-specific antigen (irradiated splenocytes with OVA323-339). After a 72 h incubation, cells were labeled with KJ1-26-Cy5.5 conjugate, which is specific for the transgenic TcR, and cell division profiles were obtained by flow cytometric analysis. Compared to FO (control), Lim (P<0.1) as well as Cur (P<0.05) diets suppressed antigen- and mitogen stimulated daughter cell division. Following PMA and Ionomycin stimulation, Lim cultures exhibited a higher percentage of cells in generation 1 (P<0.05) and a lower percentage in generation 4 (P<0.05), while Cur suppressed generation 4 (P<0.05). These results indicate that FO supplemented with limonin suppresses CD4+ T-cell division. To determine the molecular mechanism of suppression, nuclear extracts were harvested from stimulated CD4+ T-cells and NF-kB nuclear activation was assessed. The outcome of these experiments will help define the precise mechanism(s) by which limonin and curcumin alter T-cell subset activation.

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Simultaneous analysis of tocopherols, tocotrienols, phytosterols and squalene by using a gas chromatography
Kyoung-Shim Cho and Young-Sang Lee*
Soonchunhyang University, Asansi, Chungnam, South Korea, mariolee@sch.ac.kr

Vitamin E, phytosterols and squalene are important components of plant unsaponifiable fractions and their health beneficial effects are well established. In general, 8 vit. E isomers: 4 tocopherols (T) and 4 tocotrienols (T3) are analyzed by using a normal phase HPLC, while the others are analyzed by a GC. To enhance the analysis efficiency, a GC-based simultaneous determination method was developed for 13 components: alpha (a)-, beta (b)-, gamma (g)-, delta (d)- form of T and T3, squalene, campesterol, beta-sitosterol, stigmasterol, and ergosterol. For optimal separation various GC capillary columns of different polarity as well as oven temperature conditions were tested. The best GC column was CP-SIL 8 CB (25 m length, 0.25 um I.D., 0.45 um thickness) and oven temperature program was starting from 220oC to 290oC at an increasing rate of 5oC per min and then held for 28 minutes and again increased up to 300oC. Inlet and flame ionization detector temperature was 250oC and 300oC, respectively, and carrier gas (He) flow rate was 1 mL per min, split ratio was 10:1 and injection volume was 1 uL;. This method exhibited acceptable linearity (0 - 200 ng per uL), repeatability (CV 1 %) and recovery rate (> 80 %). The practicality of this developed method was proved by applying to rice bran sample and the analysis results in mg / 100 g were: 7.54 for a-T, 0.32 for b-T, 0.95 for g-T, 0.28 for d-T, 5.70 for a-T3, 0.26 for b-T3, 9.54 for g-T3, 0.44 for d-T3, 21.17 for squalene, 18.37 for campesterol, 25.16 for stigmasterol, and 68.38 for beta-sitosterol.

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Effect of pretreatments and drying temperature on the carotenoid retention of dried Brazilian squash
Mieko Kimura*, Priscila do Nascimento, Maria Aparecida Mauro
UNESP- São Paulo State University, Dept. Food Engineering and Technology, São José do Rio Preto, SP, Brasil, kimura@ibilce.unesp.br

‘Rajada Seca’ Brazilian squash is a very good and appreciated variety for processing due to its large size (30 - 40 kgs), high yield of pulp and intense orange color. This attractive color is due to the presence of carotenoids, bioactive compounds associated with adequate development and maintenance of good health, and also to the reduction of risks of degenerative diseases. Biological actions and functions of carotenoids are due to the conjugated double bond system which makes them highly susceptible to degradation (isomerization and oxidation) during processing. Among the food preservation techniques, drying is advantageous because it results in lighter and smaller products with a longer shelf-life at room temperature, thus reducing storage and distribution costs. However, it may cause flavor and color alterations as well as loss of nutrients due to exposure to oxygen and relatively high temperatures, mainly when conventional hot air drying is used. Thus, application of pretreatments aiming to improve the appearance, flavor, texture and nutrient retention, has attracted researchers’ interest over the last years. In this study, the effect of blanching and drying temperature (60°C, 70°C & 80°C) on the alpha- and beta-carotene retention of the ‘Rajada Seca’ squash was evaluated. Mature squashes were purchased in local market, peeled, deseeded, sliced (5 mm), spread on a perforated tray and dried, with and without pretreatments (blanching at 98°C/2min or osmotic dehydration with sucrose 40%/1h), in an oven with circulating air at 60°C, 70°C and 80°C. The carotenoids were extracted with acetone, partitioned to petroleum ether, saponified with 10% methanolic KOH, washed with distilled water, concentrated in a rotary evaporator, and brought to dryness with nitrogen. The dried residue was dissolved in HPLC grade acetone, filtered and injected into the liquid chromatograph (Waters model 2695) coupled to a UV-Visible photodiode array detector (Waters model 2996). The chromatographic conditions were C18 Spherisorb ODS2 column (3μm, 4.6x150mm) and mobile phase consisting of acetonitrile (0.05% TEA), methanol and ethyl acetate (60:20:20) used at a flow rate of 0.5mL/min. Identification of the peaks was done by combined analysis of the retention time, co-chromatography with pure standards and absorption spectrum. Retention was calculated taking into account the weight changes during the processes. Drying without pretreatment caused considerable losses of alpha- and beta-carotene (17-18% and 11-13%, respectively), however, significant variation in the carotenoid retention due to different temperatures was not observed. Blanching and osmotic treatment prior to drying improved the alpha- and beta-carotene retention during the process, except at 80°C where blanched sample showed lower retention. At 60°C, blanched slices showed higher retention than those pre-treated osmotically. At 70°C, retention of both pretreated samples was similar and at 80°C, osmotically pretreated slices presented higher retention of carotenoids. These results showed that two degradation processes occur during the drying of squashes, enzymatic and non-enzymatic reactions, where the former prevails at low temperatures and oxidation due to oxygen and temperature exposition prevails at high temperatures.
Acknowledgements: The authors thank FAPESP for the financial support through the project PRONEX 03/10151-4.

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Evaluation radical-scavenging properties of boiling water extract in vitro from purslane (Portulaca oleracea L.)
Yuanhu Zhang*, Lingling Li, Xin Chen, Jianping Shao, and Yaqin Li
College of life science, Shandong Agricultural University, Taian 271018, Shandong province, P.R.CHINA, yyhzhang@sdau.edu.cn

The antioxidant activity of puralane (Portulaca oleracea L.) extracts was tested by hydroxyl radical-scavenging, superoxide anion radical-scavenging, DPPH radical scavenging and ABTS radical scavenging methods. Total phenolic compounds and flavonoids were also detected. The results showed that the highest antioxidant activity and polyphenols was in root extracts, the lowest in stem, antioxidant activity decreased in the order of root > flower > leaf > whole plant > stem. Radical -scavenging capacity of boiling water extract from purslane was higher than that of extract with non-heated water, the optimal boiling water extraction time was 15 minutes. There was a strong relationship between the content of flavonoids, total phenolic compounds and antioxidant activity of purslane extracts. It indicated that the boiling water extract from Portulaca oleracea L. could serve as a rich potent source of antioxidants.

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Indigenous leafy vegetables in the diet and agriculture of the sandawe of Tanzania
Matthew C. Knisley*1 and Agnes Nyomora2
1606 Bluff Street, Greenfield, IL, USA; mcknisley@gmail.com; 2University of Dar es Salaam, Department of Botany, PO Box 35065, Dar es Salaam, Tanzania

Few studies address the use of leafy indigenous vegetables in poorly understood contexts, such as emergent farming communities. This quantitative and qualitative ethnographic study analyzes the role of indigenous leafy greens in the diet and agriculture of the Sandawe, a former hunting and gathering ethnic group located in Kondoa District, Tanzania, that began transitioning to an agricultural subsistence base approximately 100 years ago. Availability and dietary importance of indigenous vegetables were determined by surveying transects through areas of human activity, identifying and weighing harvested greens, observing the preparation of greens, and compiling food logs. Interviews, participant observation, and mapping provided insights into Sandawe planting schedules, field and garden configuration, seed saving practices, and maintenance of cultivated areas. Attitudes towards the growth and consumption of greens and local folk taxonomies of greens were constructed through the use of interviews and pile sorting. While decreasing plant biodiversity in both wild and agricultural settings has compromised food security in many settings, the use of leafy greens among the Sandawe is high. Linguistic and ecological evidence suggests that many leafy greens commonly consumed by the Sandawe accompanied the agricultural complex. This research illustrates the importance of documenting cultural norms and individual preference as they relate to the maintenance of biodiversity and could serve as a model for future quantitative studies of indigenous leafy vegetables.

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Ascorbic acid content and antioxidant capacity of the acerola cherry (Malpighia punicifolia L)
Tara Rimstad2, Charles Chun Hu*1, David D Kitts2
1Naturally Nova Scotia Health Products Limited, 2University of British Columbia, Canada, charles.chunhu@gmail.com

The ascorbic acid content and antioxidant activity of acerola cherry (Malpighia punicifolia L) extracts was examined using freeze dried acerola fruit collected at three maturity stages. Vitamin C content was analyzed using HPLC, and, in vitro antioxidant activity was assessed using the oxygen radical absorbance capacity (ORAC) and Trolox equivalent absorbance capacity (TEAC) assays. Both maturation stage of the fruit and duration of exposure to heat and humidity (55oC and 80% relative humidity) greatly affected the ascorbic acid content and antioxidant activity. Ascorbic acid content decreased with maturation from 254.3 to 90.50 mg/g dry weight (dw), while mature fruit subjected to 12 days exposure to heat and humidity had a reduced ascorbic acid content (e.g. 13.69 mg/g dw). Correspondent to the decrease of ascorbic acid content, the antioxidant capacity of the acerola fruit was also lower as the fruit matured or when exposed to heat and humidity. The antioxidant activity of immature, unexposed fruit was attributed mostly to the ascorbic acid content. With maturation and exposure to heat and humidity, results of ORAC assay suggested that other possible antioxidant components existed in the fruits when they were ripe. Intracellular oxidation model with Caco-2 cells revealed that acerola cherry extracts suppressed peroxyl radical induced intracellular oxidation. The antioxidant activity of acerola fruit was found to be strong compared to other fruits, especially for immature and partially mature acerola. Therefore, acerola cherry can be used as a good source of natural ascorbic acid and antioxidant with potential application in functional food and food supplements.

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Cloning and sequencing of PGIP from ‘Jin Series’ almond (Prunus dulcis)
Yanxia Wu1, Xinping Fan2, Xiaoyu Yang2, Qiufen Cao3, and Jian-bao Tian*2
1College of Horticulture, Shanxi Agricultural University, 030801 Taigu, Shanxi,China; 2Institute of Pomology, Shanxi Academy of Agricultural Sciences, 030815 Taigu, Shanxi, China; 3Center of Biotechnique Research, Shanxi Academy of Agricultural Sciences, 030015 Taiyuan, Shanxi, China, tianjb-001@163.com

Specific primers synthesized according to conservative regions of polygalacturonase inhibiting protein (PGIP) gene were used to amplify Prunus dulcis genomic DNA by polymerase-chain reaction (PCR). Six bands (pgip1, pgip2, pgip3, pgip4, pgip5 and pgip6) of genes were obtained and cloned into PBS-T vector. According to the length of bands, 717bp, 864bp, 796bp were A1 (pgip1, pgip2, pgip3), A2 (pgip4), A4 (pgip5, pgip6) respectively. DNA sequences showed that the fragments taken together were the gene encoding PGIP. A2 and A3 contained two exons interrupted by one intron, which has GT-AG sequence. Its DNA and amino acid sequences were highly homologies to those from Prunus persica, Prunus salicina, Prunus americana, and Prunus mume respectively. A conserved lencinerial fragment exists in the derived protein sequence.

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Lemon (Citrus lemon L. Burm) as a source of unique bioactive compounds
Jin Hee Kim*, G. K. Jayaprakasha and Bhimanagouda S. Patil
Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, Texas A&M University, College Station, TX, USA, jinpi@neo.tamu.edu

Citrus contain several bioactive health maintaining compounds which may have potential for prevention of certain diseases. In order to understand the role of these bioactivities, isolation and characterization are critical steps of bioactive driven studies. The lemon seed powder (7.4576kg) was extracted with different solvents, such as hexane and ethyl acetate (EtOAc) using Soxhlet extractor. The EtOAc extract was concentrated by rotary evaporation and dried. The dried residue was extracted with dichloromethane (DCM) and concentrated to obtain limonin crystals (13.7506g). However, other four compounds such as obacunone (0.0683g), nomilin (3.6678g), β-sitosterol glucoside (SG, 0.1762g) and limonexic acid (LNA, 0.3404g) were obtained by silica gel column chromatography. These compounds were detected by thin layer chromatography and purity was confirmed by high-performance liquid chromatography. Furthermore, the structures of the isolated compounds were confirmed by atmospheric pressure chemical ionization –mass spectrometry. To the best of our knowledge isolation and characterization of obacunone, nomilin, SG, and LNA from lemon seeds using combination of chromatographic techniques was achieved for the first time. Further studies on potential of these compounds on cancer prevention using cell culture are in progress. This project is funded by the USDA-CSREES # 2006-34402-17121 “Designing Foods for Health” through the Vegetable & Fruit Improvement Center.

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Post harvest control of aflatoxin in pistachio
Ebrahim Hadavi* and Noushin Ghazijahani
Agriculture Faculty of Islamic Azad University, Karaj Campus, Postal Code 31876-44511 , Eram Blvd. Mehr-Shahr, Karaj, Iran, hadavi@kiau.ac.ir

Aflatoxin concentration in Iranian orchards is evaluated. Based on high contamination observed if effective measure(s) is not applied the resulting risk remains high. Efficacy of different processing stages on removal of contaminated nuts is evaluated and some recommendations are given for better designing of processing machinery. After processing presence of bright greenish yellow fluorescence (BGY-F) could be used as a discriminating factor for identification of highly contaminated nuts. Mean concentration of aflatoxin in BGY-F portion was 158 ppb against 2.2 ppb in non-BGY-F portion. In these samples by removing mean of 4.4 percent BGY-F nuts average of 82 percent of aflatoxin is excluded as well. As storage of nuts in poorly ventilated conditions after harvest causes considerable increase in share of BGY-F nuts, Percent of BGY-F nuts could be used in any subsequent stage as an Index for quality assessment of post harvest handling as a pricing factor in pistachio. The BGY-F percentage have been rised from 4 percent in properly handled nuts up to 21 percent in poorly handled nuts.

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Steroidal saponin from the roots of momordica charantia
Kumud Shrivastava
Department of Chemistry, Govt. MLB Girl’s P.G. Autonomous College, Barkatullah University, BHOPAL (M.P.)- India, dp_shrivastava@yahoo.com

The roots of Momordica charantia were found to contain a steroidal saponin by combined physico-chemical studies and identified as; stigmast-5, 17(20)-dien-3-O-β-D-arabinofuranosyl (1 → 4)-O-The roots of Momordica charantia were found to contain a new steroidal saponin by combined physico-chemical studies and identified as; stigmast-5, 17(20)-dien-3-O-b-D-arabinofuranosyl (1 → 4)-O-a-L-rhamnopyranosyl (1 → 4)-O-b-D-glucopyranoside. Momordica charantia (N.O- cucurbitaceae) is known as “bitter guard” in English and also cultivated as a vegetable crop. The fruits and leaves are reported to be useful in piles, leprosy, juindice and as bermifuge. The juice of the leaves is also reported to be emetic purgative and given in bilious infections and useful in curing burning of the soles of the feets. The roots are as tringent, useful in haemorrhoids. The juice of the fruits used in snake bite. Juice of fruits is also useful for diabetees.
-L-rhamnopyranosyl (1 → 4)-O-β-D-glucopyranoside. Momordica charantia (N.O- cucurbitaceae) is known as “bitter guard” in English and also cultivated as a vegetable crop. The fruits and leaves are reported to be useful in piles, leprosy, juindice and as bermifuge. The juice of the leaves is also reported to be emetic purgative and given in bilious infections and useful in curing burning of the soles of the feets. The roots are as tringent, useful in haemorrhoids. The juice of the fruits used in snake bite. Juice of fruits is also useful for diabetees.

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A polygalacturonase inhibitor protein extracted from Pink Lady apple may provide some defence against Colletotrichum acutatum infections
R. Gregori1, M. Mari1, P. Bertolini1, J.A. Saudo-Barajas2, J.B. Tian*3 and J.M. Labavitch4
1CRIOF Department of Agri-Food Protection and Improvement, University of Bologna, Italy, 2Centro de Investigaciones en Alimentaciony Desarrollo A.C., Unidad Culiacan, Sinaloa, Mexico, 3Pomology Institute, Shanxi Academy of Agricultural Science, 030815 Taigu, Shanxi, China, 4Department of Plant Science, University of California, Davis, CA, USA, tianjb-001@163.com

Polygalacturonases (PGs) have been shown to be an important pathogenicity factor for soft rot fungi. PGs play an important role in cell wall degradation and they probably interact with other cell wall modifying proteins in breaking down the cell wall. Colletotrichum acutatum, a fungal pathogen of fruit, produced PGs when grown on pectin or apple cell walls. A PG-inhibiting protein (PGIP) was extracted from apple, cv Pink Lady, fruit that had been stored at 0oC for 4 months. PG-inhibiting activity of the extracts was tested in vitro and in vivo against an endo-PG of C. acutatum (EC 3.2.1.15). In vitro inhibition was 47.5% while inhibition in vivo ranged from 33.9% to 54.4% after incubation for 4 days at 20oC.

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Characterization of health-promoting Polymethoxylated flavones from Jamaican and Mexican citrus peel
Curtis O. Green1, Andrew O. Wheatley*1, 2, Anthony U. Osagie1, Lowell L. Dilworth1, Errol Y. St. A. Morrison1, Helen N. Asemota1, 2
1Department of Basic Medical Sciences and 2Biotechnology Centre, University of the West Indies, Mona Campus, Kingston 7, Jamaica W.I., andrew.wheatley@uwimona.edu.jm

The polymethoxylated flavones (PMFs) are flavonoids that are almost exclusively found in Citrus peel and display potent anti-hypercholesterolemic, anti-cancer and anti-atherosclerotic properties. The aim of the present study was to characterize the polymethoxylated flavones (PMFs) in Jamaican and Mexican citrus peels by high-performance liquid chromatography. The PMFs were extracted from sun dried citrus peels with reagent grade methanol. Analyses were carried out by reverse-phase HPLC and UV detection. Our findings indicated that Jamaican and Mexican Citrus peels contained appreciable quantities of the PMFs- the Jamaican Ortanique peel contained the highest quantity of the PMFs (34,393 ± 272 ppm), followed by Tangerine (28,389 ± 343 ppm) and Mexican Sweet Orange (21,627 ± 494 ppm). Jamaica produces Citrus fruits on a large scale which generate large quantities of Citrus peels and by-products that are generally discarded or under-utilized. These Citrus peels and other by-products of citrus processing could therefore be used to generate relatively large quantities of PMFs that could serve as food additives of natural origin or as pharmaceutical and nutraceutical supplements for ameliorating metabolic disorders that are common in the Caribbean.

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Evaluation of antioxidant activity in some American tropical fruits (acerola, acai, mangaba and uvaia) by DPPH method
.Maria S.M. Rufino1,. Edy S de Brito*, Ricardo E Alves2, Fulgencio D. Saura-Calixto3, Jara Jimenez-Perez3
1.UFERSA/BR 110 - Km 47, Bairro Pres. Costa e Silva, Mossoro-RN, Brasil, 2. Embrapa Agroindustria Tropical/Planalto do Pici, Fortaleza-CE, Brasil, 3. Instituto del Frio, Madri, Espana, edy@cnpat.embrapa.br

Currently, great emphasis is given to the activity of natural antioxidants present in fruits and vegetables. These compounds could reduce the oxidative stress level, which can disturb the equilibrium of the antioxidant/pro-oxidant system on intact cells, resulting oxidative damage on lipids, proteins, carbohydrates and nucleic acids. Fruits are well known natural products rich in antioxidant compounds and by this way are in a consumers increasing preference attitude. There is a plenty of methodologies to evaluate the Total Antioxidant Activity (TAA), being the DPPH (2,2-Diphenyl-1-picryl-hidrazil) radical scavenge one of the most important. On this method the antioxidant efficiency is measured at ambient temperature, avoiding the thermal degradation risk of the antioxidant compounds. However, the reaction mechanism depends on the antioxidant structural conformation and consequently the reaction velocity may be changed. This research has the aim of evaluate the kinetic of DPPH capture by some tropical fruits (acerola, mangaba, acai e uvaia) extracts. The fruits were harvested on the Ceará and Piauφ states and transported to the Post-Harvest Technology and Physiology Laboratory from Embrapa Agroindustria Tropical, where the capacity to reduce 50% the DPPH concentration of different fruit extracts was evaluated. The absorbance decrease until stabilization was followed at 515 nm. Reaction stabilization happened at 10 minutes for acerola, 30 minutes for mangaba, and 120 minutes for aτaφ and uvaia, which demonstrated the importance of the estimation of stabilization time for each sample. Reaction stabilization time was also important to achieve a better knowledge of the mechanisms of each fruit TAA. Acerola presented the highest TAA (670 g/g DPPH) followed by uvaia (3246 g/g DPPH), mangaba (3385 g/g DPPH) and acai (3778 g/g DPPH). Antioxidant potential of this fruits could be attributed to different compounds evaluated, like vitamin C (1357mg/100g) and phenolics (1063 mg/100g) in acerola. Different TAA as well as different stabilization times reinforces the importance of a diversified diet in a way to obtain the better from each fruit. (Support: CAPES, CNPq e EMBRAPA)

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Evaluation of antioxidant activity acerola clones by different antioxidant methods
Caroline de Goes Sampaio1, Maria do Socorro Moura Rufino2, Ricardo Elesbπo Alves3, Edy Sousa de Brito*3, Selene Maia de Morais1
1UECE, Fortaleza, CE, Brazil, 2UFERSA, Mossor≤, RN, Brazil, 3Embrapa Agroindustria Tropical, R Dra Sara Mesquita, 2270, Pici, 60511 110, Fortaleza, CE, Brazil, edy@cnpat.embrapa.br

Acerola is a fruit with a high nutritional value not only due to its vitemin C content, but also to beta-caroten and anthocyanins. the oxidative stress is one of the main mechanisms responsible for cell and tissue damage, related to health deseases. This stress can be controled through food containing naturally antioxidants. The aim of this work was to evaluate the quality and the total antioxidant capacity of six commercial acerola clones. Fruits were analyzed for total soluble solids (TSS), total soluble solids, total tritatable acidity (TTA), pH, TSS/TTA, vitamin C, anthocyanins, yellow falvonoids, total carotenoids, total extractable polyphenols and total antioxidant activity (DPPH, ABTS, FRAP). Chemical evaluation highletd the II 47/1 clone due to its high vitamin C and anthocyanin content. Acerola clones presented an average IC mean of 697.1, with the clone II 47/1 presenting the higher IC50 (461.5 g/g pulp). For ABTS method the clones Cereja and II47/1 presented the higher values 158.7 e 138.3 uM Trolox/g pulp, respectively. It was also observed a significant difference among samples. The same was observed for FRAP method, with the clone Sertaneja presenting the higher value (306.9 uM/g pulp).

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Determination of bioactive compounds and antioxidant activity on different aτaφ (Euterpe oleracea Mart) progenies
Marcela Coelho de Souza1, Ricardo Elesbπo Alves2, Edy Sousa de Brito*2, Raimundo Wilane Figueiredo1, Carlos Farley Herbster Moura2
1Departamento de Tecnologia de Alimentos, Universidade Federal do Cearß, Fortaleza, CE, Brasil. 2Embrapa Agroindustria Tropical, Fortaleza, CE, Brasil, edy@cnpat.embrapa.br

Nowadays, there is a great interest on aτaφ (Euterpe oleraceae Mart) fruits due to its functional and energetic properties. Based on their chemical characteristics aτaφ fruits from different progenies were evaluated in order to select genetical material of superior antioxidant activity. The fruits were collected from eight progenies of the the germplasm bank from Embrapa Agroindustria Tropical, located at Paraipaba, CE. Each sample was evaluated for ascorbic acid, carotenoids, yellow flavonoids, anthocyanins, total extractable polyphenols, condensed tannins and antioxidant activity by the carotene/linoleic acid bleaching method. Regarding the majority of parameters evaluated aτaφ fruits presented a high variation among progenies. It was observed an average ascorbic acid mean of 58.7 mg/100 g. Anthocyanin avarage was 108.9 mg/100g, with variations from 73.52 to 143.52 mg/100g, and progenies P131-3 (143.62mg/100g) and P034-3 (134.71 mg/100g) presented the highest contents. In general, a high content of all biological active compounds was observed. Aτaφ extracts presented an excellent oxidation inhibition, with progenies presenting different antioxidant capacities. Antioxidant activity varied from 47.7 to 75.0 % of oxidation inhibition (% OI), with higher values for progenies P049-6 (75.0 % OI), P048-8 (72.0 % OI), P032-5 (70.0 % OI) and P308-2 (67.0 % OI).

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Bioactive compounds and antioxidant activity of cashew peduncle from different early dwarf plants (Anacardium occidentale L.)
Cyntia Rafaelle Amaral de Abreu1, Ricardo Elesbao Alves2, Carlos Farley Herbster Moura2, Geraldo Arraes Maia1, Edy Sousa de Brito*2
Departamento de Tecnologia de Alimentos, Universidade Federal do Ceara, Fortaleza, CE, Brasil1. Embrapa Agroindustria Tropical, Fortaleza, CE, Brazil2, edy@cnpat.embrapa.br

Fruit antioxidant activity is related to compounds such as vitamins and phenolics, which play an effective role on free radical scavenge. Such activity varies among species and cultivars. The aim of this work was to evaluate and quantify antioxidant compounds on cashew peduncles from different early dwarf clones. The following analysis were done: vitamin C, total carotenoids, total anthocyanin, yellow flavonoids, total phenolics, condensed tannins and total antioxidant activity. The peduncles were harvested at the Embrapa’s experimental station at Pacajus, CE, Brazil. Regarding vitamin C and total carotenoids the clone BRS 189 presented the highest contents, with values of 270.03 mg/100g and 0.93 mg/100g, respectively, however the vitamin C content were not statiscally different to BRS Bahia 12 (254.34 mg/100g). The highest values of total anthocyanin and yellow flavonoids were founded in clones BRS 265, Embrapa 51 and BRS Bahia 12, which had 13.10 mg/100g for total anthocyanin and 61.23 mg/100g for yellow flavonoids. Total phenolics presented the greatest variability amongst samples, with values from 99.53 mg/100g to 236.97 mg/100g. The same was observed for antioxidant activity with an average mean of 16.35 µmol trolox/g of fruit pulp. For condensed tannins the BRS 265 clone had the highest value (11.23 mg/100g), while Embrapa 51 had the lowest (5.67 mg/100g).

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Antioxidant capacity and phenolic content of stingless bee honey from Amazon in comparison to Apis honey
Deborah H. Bastos1, Michele C. M. Dos Santos2, Simone Mendona3, Elizabeth A.F.S. Torres4
1Brazil Sao Paolo School of Public Health USP, 2Sao Paolo FAPESP, 3Sao Paolo CNPq

Many fruits and vegetables are good sources of natural antioxidants because of their phenolic contents. Similarly, honey have shown in its composition phenolics compounds such as flavonoids, that present a high antioxidant potential. In sensorial aspects this honey is quite different from Apis (european bee) honey. Due to biological and pharmacological activities, it has been extensively used in folk medicine as an improver of digestive and respiratory systems, fertility, skin and visual disorders. Besides sugar and water, other minor compounds play an important role in flavor and biological properties. Phenolic compounds are responsible for the antioxidant activity and are also used as biomarkers of Apis honey origin. Up to date there are no data concerning phenolic compounds composition and antioxidant activity of stingless bee honey from brazilian amazon region. Objectives: Assess soluble solids (Brix), quantify total phenolic compounds (TPC) and evaluate in vitro antioxidant activity of Meliponinae methanolic extract in comparision to Apis methanolic extract of different floral sources: orange tree (Citrus sinensis) and eucalyptus (Eucaliptus spp) Material and Methods: Five samples of stingless bees (Four of Melipona seminigra merrilae bee (MSM) and one of Melipona compressipes manaosensis bee (MCM)) honey and two samples of Apis Mellifera bee honeys of different floral source (orange tree (AMO) and eucalyptus (AME)) were obtained directly from producers. The phenolics compounds were extracted with methanol on Amberlite XAD-2 resine. The soluble solids were measured through refratometry. The TPC and flavonoids were assayed on the methanolic extract using Folin-Ciocalteau and addition of AlCl3 methods, respectively Antioxidant capacity was determined through scavenging DPPH free radical and expressed as IC50(Capacity of inhibition 50% of free radical DPPH). Results: The Brix varied from 67 to 79 in MSM, 75 in MCM, 80.5 AMO and80 AME honeys. The TPC varied from 458,65 (sd 0,64) to 1329,4 (sd 121,8) mcg/100 g/ QE (Quercetin Equivalent) in Meliponinae honeys and 1859,70 (sd 206,36) to 5673,25 (sd 195,45) mcg/100 g/ QE in Apis honeys and the flavonoids content varied from 74,79 (sd 21,26) to 203,19(sd 58,66) mcg/100 g/ QE in Meliponinae honey and 498,16 (sd 1,43) to 1725,21 (sd 56,87) g/100 g/ QE in Apis honey.The IC50 varied from 26,34 to 5,89 mcg/100 g/ QE in Meliponinae species and 9,33 to 9,38 mcg/100 g/ QE and compared to BHT (6,32 mcg/ml). Conclusion: Although Apis honeys presented higher value for TPC and flavonoids than stingless bees, one of Meliponinae species presented better antioxidant activity than all other samples analyzed. This pointed to need of more studies to identify the structure of phenolics of honey samples of Brazilian bees species that present antioxidant potential even in lower concentration.

were assayed on the methanolic extract using Folin-Ciocalteau and addition of AlCl3 methods, respectively Antioxidant capacity was determined through scavenging DPPH free radical and expressed as IC50(Capacity of inhibition 50% of free radical DPPH). Results: The Brix varied from 67 to 79 in MSM, 75 in MCM, 80.5 AMO and 80 AME honeys. The TPC varied from 458,65 (sd 0,64) to 1329,4 (sd 121,8) mcg/100 g/ QE (Quercetin Equivalent) in Meliponinae honeys and 1859,70 (sd 206,36) to 5673,25 (sd 195,45) mcg/100 g/ QE in Apis honeys and the flavonoids content varied from 74,79 (sd 21,26) to 203,19(sd 58,66) mcg/100 g/ QE in Meliponinae honey and 498,16 (sd 1,43) to 1725,21 (sd 56,87) g/100 g/ QE in Apis honey.The IC50 varied from 26,34 to 5,89 mcg/100 g/ QE in Meliponinae species and 9,33 to 9,38 mcg/100 g/ QE and compared to BHT (6,32 mcg/ml). Conclusion: Although Apis honeys presented hi
gher value for TPC and flavonoids than stingless bees, one of Meliponinae species presented better antioxidant activity than all other samples analyzed. This pointed to need of more studies to identify the structure of phenolics of honey samples of Brazilian bees species that present antioxidant potential even in lower concentration."

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Phenolic Compounds and Antioxidative Activity of Clear and Cloudy Juices Produced From Selected Scab Resistant Apple Cultivars
Dr. Jaroslaw Markowski, M.Sc. Monika Mieszczakowska, Prof. Witold Plocharski
Department of Storage and Processing, Research Institute of Pomology & Floriculture, Skierniewice, Poland, witold.plocharski@insad.pl; jaroslaw.markowski@insad.pl

Experiments were performed to investigate effect of raw material and technology on phenolics composition of clear and cloudy apple juices. Six cultivars from a batch of 27 scab resistant cultivars were selected for experiments, based on preliminary composition analysis and sensory trials. The phenolics composition of apples and juices was determined by HPLC. Antioxidative properties of obtained products were determined as well using ABTS 50% method. The following compounds were identified and quantified: flavan-3-ols, dihydrochalcones, hydroxycinnamic acids, quercetin and quercetin glycosides. The most abundant in phenolics was ‘Melfree’, exceeding scarce ‘Rajka’ 2,2 times. Procyanidins dominated in most cultivars, however chlorogenic acid dominated in ‘Early Freegold’. Also in both kinds of juices chlorogenic acid predominated, however its amount in cloudy juices was on average 2,4 times higher than in clear juices. Generally, phloretin xyloglucoside and p-coumarylquinic acid were most stable during enzymatic treatment of the mash and clarification. The retention of polyphenols during juice production was cultivar dependent. In ‘Novamac’ juices, either cloudy or clear, none of the investigated phenolic compounds was retained in more than 40% compared to raw material, whereas in ‘Melfree’ in cloudy juice retention of catechin, epicatechin, phloretin xyloglucoside, chlorogenic acid and p-coumarylquinic acid was very good: 80%, 71%, 70%, 86% and 100% respectively. In clear ‘Melfree’ juice retention of the above mentioned compounds was 0%, 16%, 43%, 36%, and 84% respectively. Higher retention in cloudy juice may results from application of small amount of ascorbic acid (200 mg/kg of apples) during fruit disintegration to protect material from oxidative action of native enzymes, and shortening processing cycle to minutes compared to clear juice production. Lower phenolics content in clear juices may also be due to pectolytic enzymes used to increase juice yield, which always have some “accompanying activity”. Higher content of phenolics in cloudy juices contributed to much higher antioxidative activity of these products compared to clear juices. Determination coefficient between ABTS 50% values and antioxidative activity was 0, 9194. Results obtained indicate that cloudy juices in general contained higher amount of phenolics than clear juices and were characterized by higher antioxidant activity. Mash depectinisation and clarification of juices decreases biological value of apple juices.
Results were obtained within „ISAFRUIT” Project,
financed by the European Union - Contract No. FP6-FOOD 016279-2