Posters
Ann Nutr Metab 2013;63(suppl 1): 1-1960

T6. Functional Foods and Bioactive
Compounds

PO2658
EVALUATION OF DEFATTED SOY BASED FORMULAS PREPARED FOR INFANTS AND YOUNG CHILDREN WITH ACUTE DIARRHEA
S. Zaki1, M. Abou-Zekri2, A. El-Akel1, L. El-mahdi3, N.
Hassan4
1
Department of Food Technology,Faculty of
Agriculture,Cairo University,Giza, Egypt
2
Department of Endemic Diseases, Abou Elrish Children Hospital, Cairo university, Cairo, Egypt
3
Department of Horticulture Crops, Food Technoloy
Institute, Agriculture Research Center, Ministry of
Agriculture,Giza, Egypt
4
Department of Experimental Kitchen, Food Technology Institute, Agriculture Research Center,
Ministry of Agriculture,Giza, Egypt
Background and objectives: Recent data supported that
diet composition introduced to infants and young children
during diarrheal episodes is controversial so it has been the
concern of extensive research. Study aimed to formulate and
evaluate some proposed formulas for this group during acute
diarrhea.
Methods: Six different defatted soy based formulas with
yellow carrots, rice, potato starch powder and orange were
prepared. Outpatients’ infants and young children (140) with

acute diarrhea, visiting the Center for Social and Preventive
Medicine, Cairo University Children’s Hospital, Egypt, were
enrolled in the study. Participants were randomly assigned to 7
groups, 20 each. The control group did not receive any dietetic
intervention. At the first visit, participant’ mother in the other
groups randomly received a formula at in a powder form to
be dissolved and used at home. Patients were followed up for
2 weeks in two visits (after 5 and 15 days of enrollment) and
assessed for diarrhea duration, number of motions/day, stool
consistency, weight gain and their acceptability of the formulas.
Statistical analysis of the results was performed.
Results: Feeding patients formulas demonstrated insignificant (p< 0.05) differences among age, gender, and illness cha© 2013 S. Karger AG, Basel
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Published online: September 13, 2013

racterize of cases. The percentages of cases with watery, loose
and semi solid stools were greatly reduced after five days in all
the groups. Insignificant difference (p< 0.05) was found among
weights of formulas’ groups and control at enrollment, after 5
and 15 days. These groups showed significantly lower mean
number of motions per day after getting proposed formulas for
5 dayscompared to control group (p<0.001).
Conclusions:During diarrhea episodes, feeding participants proposed formula showed better outcome than those not
receiving dietetic intervention.
Key words: acute diarrhea, infant, young children, soy based formulas.

PO2659

DEVELOPMENT OF PROBIOTIC FOOD MULTIMIX
(PFMM)
R. Djoulde Darman1
1
Department of Agriculture Husbandry and By products, The Higher Institute of the Sahel, University
of Maroua, Maroua, Cameroon
Background and objectives: The food multimix (FMM)
concept is one of the attempts used successfully by international programmes to meet the food insecurity challenges in
poor African rural area. Despite its success the current FMM
formulation concept target mainly common nutrient as carbohydrates, protein and vitamins thus lacking some bioactive
compounds. This paper aims at adding live Bifidobacterium to
a FMM formulation to develop a FMM enriched with bioactive
compound.
Methods: The FMM were developed using fermented
sorghum flour, cowpea, tiger nuts milk, red Hibiscus extracts
and Aloes vera extracts. The probiotic were a strain ofBifidobacterium Longum isolated from chocolate bars. The FMM
were first prepared and aliquot (102UFC/g to 1010UFC/g) of
live encapsulated bifidobacterium added. The mixture allowed
drying in an owen. The Arrhenius model supplemented by the
linear model was used to predict the survival time of bifidobacterium in PFMM stored at ambient conditions.
Results: The proximate composition of developed PFMM
shows an energy value of 427.77±23 Kcal/100g, a protein value
of 23.4±6g/100g, and total fats value of 6.29g/100g. Vitamins
iron and calcium were also recorded. The survival time experiAnn Nutr Metab 2013;63(suppl 1):1–1960

1551


ments indicate an optimal rate of bifidobacterium as 106UFC/g
with 30% death after 6 month storage time.

Conclusions: The new probiotic food multimix can be considered as a powerful bioactive enriched meal, with adequate
required energy and key nutrients together with live microroganisms for bioactives compound.
Key words:Probiotics, Food multimixt, North Cameroon

PO2661
BENEFITS OF HYDROLYSED COLLAGEN SUPPLEMENTATION IN SKIN’S PHYSIOLOGY OF MIDDLEAGED PORTUGUESE WOMEN
N. Tavares1, C. Rosado1, L. Rodrigues1, S. Abreu1,2
1
CBIOS, Escola de Ciências e Tecnologias da Saúde,
Universidade Lusófona, Lisboa, Portugal
2
Research Centre In Physical Activity, Health and
Leisure, Faculty of Sport, University of Porto, Porto,
Portugal
Background and objectives:Collagen play a key role in
normal dermatologic functioning. However, very few is known
about the effects of hydrolysed collagen supplementation on
skin’s functions.We evaluated the effects resulting fromhydrolysed collagen intake in skin’s water profile and biomechanicsin
mature women.
Methods:Using data from measurements of transepidermal
water loss, epidermal capacitanceand biomechanical behaviour, before and after 30 days of supplementation we evaluated effects on skin’s physiology in 18 women, 52.3y (SD±7.3).
Volunteer’s hydration status was assessed by plasma osmolality.
Supplementation consisted in 2,5 g hydrolysed collagen or placebo each day for 30 days. A randomized, placebo-controlled,
double-blind trial was chosen.
Results: The marker of hydration status was within the
range of normalcy for all women. Hydrolysed collagen supplementation significantly increased skin hydration (p=0.000)
and elasticity (p=0.000). Significant variation in transepidermal water loss could not be observed.
Conclusions: In the present experimental conditions, a 2,5
g per day oral hydrolysed collagen supplementation improved
skin hydration and elasticity.

Key words: Collagen hydrolysate, supplementation, skin,
women

PO2662
DEVELOPMENT OF TASTY BREAD CONTAINING
FUNCTIONAL ULVA PERTUSA POWDER
T. Yamauchi1, A. Koide1, K. Matsumoto2, K. Mamiya1, T.
Banno2, A. Yamamoto3, M. Moriyama4
1
Faculty of Human Life and Environmental Sciences,
Nagoya Women’s University, Nagoya, Aichi, Japan
2
Department of Applied Life Studies, College of
Nagoya Women’s University, Nagoya, Aichi, Japan
3
Department of Food Science and Nutrition, Aichi
Gakusen College, Okazaki, Aichi, Japan
4
Faculty of Human Life and Environmental Sciences,
Aichi Gakusen University, Okazaki, Aichi, Japan
Background and objectives: Ulva pertusa (Sea lettuce) has
not been used as food in Japan due to its tough fibrous texture;
however, in light of recent findings that the seaweed contains
high levels of iron, calcium, vitamins, and polyphenols, its potential food applications are currently drawing increasing attention. We attempted to develop tasty loaf bread, which is the
most regularly consumed type of bread in Japan, containing
ulva pertusa powder to utilize the functional components of
the plant.
Methods: Bread samples were prepared to contain powdered ulva pertusa (particle size, 250 ƒÊm) at levels of 0 (control),
1, 2, and 3%, and examined for the contents of vitamin C, chlorophyll, and polyphenols, along with DPPH radical-scavenging
activity, specific loaf volume, color, texture (using a creep meter), and sensory attributes (on a five-point scoring system).

Data were analyzed by Tukey-Kramer’s test (p<0.05).
Results: The contents of vitamin C, polyphenols, and chlorophyll, and the level of DPPH radical-scavenging activity increased with the amount of added ulva pertusa powder. The
breads containing 2 and 3% ulva pertusa powder showed a
significantly lower specific loaf volume, a significantly higher
hardness value, and a significantly lower cohesiveness value
than the control. In relation to the amount of added ulva pertusa powder, there was a significant reduction in lightness and
a significant increase in greenness and yellowness. The bread
containing 1% ulva pertusa powder received significantly higher scores in the sensory evaluation than the other breads.
Conclusions: The contents of the functional components
increased with the amount of ulva pertusa powder added, and
the bread containing 1% ulva pertusa powder was ranked first
in terms of preference. Regular consumption of ulva-pertusacontaining bread could be an effective way to increase the intake of its functional components.
Key words: ulva-pertusa-containing bread, functional
components, color, texture, preference

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Ann Nutr Metab 2013;63(suppl 1):1–1960

Abstracts


PO2663
ANTIOXIDANT ACTIVITY AND PHENOLICS CONTENT OF SWEET CHERRIES (PRUNUS AVIUM L.)
FROM NORTHERN SERBIA
M. Popovic1, D. Prvulovic1, D. Malencic1, M. Ljubojevic1,
G. Barac1, V. Ognjanov1, B. Popovic1
1
Faculty of Agriculture, University of Novi Sad, Novi
Sad, Serbia

Background and objectives: Sweet cherry is a valuable source of some bioactive compounds important for human health
preservation. Recent increasing interest in nutraceuticals and
functional foods has led plant breeders to initiate selection of
crops with higher antioxidant activity. Red fruits, including
sweet cherries, have been reported to contain various phenolics
which contribute to total antioxidant activity. Thus, the aim of
this study was to determine the fruit content of different phenolic compounds and to estimate their antioxidant activity.
Methods: Red-colored fruits of 15 cultivars were collected from the productive orchard “Sloga” at Ka, in vicinity of
Novi Sad. The plants were growing under continental semiarid agroecological condition typical for the Panonnian plain
lowlands representing the north of Serbia. The content of total
polyphenolics (TP), total tannins (TT), total flavonoids (TF)
and anthocyanins (A) has been determined in 70% ethanol
fresh fruit extracts. In addition, the DPPH and FRAP antioxidant assays were evaluated, as well.
Results: TP content in the sweet cherries ranged from 7.45147.31 mg GAE 100-1 g fresh weight (f.w.). Cv. Peter had the
highest TP and TT content, followed by Katalin (137.55 mg
100-1 g) and Carmen (126.90 mg 100-1 g). The cultivars with
high TF content observed were Peter, Rita and Margit with
41.80, 39.00 and 36.98 mg rutine g-1 f.w., respectively. Sweet
cherries cultivars abundant in TP, TT and TF also contained
more anthocyanins. The highest content was in the Katalin
fresh fruits, 0.69 mg cyanidin-3-glycoside g-1 f.w. The DPPHvalues for investigated extracts varied in a wide range between
39.76% and 78.71%.
Conclusions: DPPH activity was found to have a positive
correlation with TP (0.771), TT (0.747) and A (0.548). However, the correlations of FRAP values with phenolics were relatively lower.

PO2664
PROTECTIVE EFFECT OF LYCOPENE AGAINST
ACUTE MYOCARDIAL INFARCTION
Y. Zhang1, H. Wang2, Y. Hou3, Y. Yang2, H. Pan1
Department of Nutrition and Food Hygiene, Public

Health School of Harbin Medical University, Harbin,
China
2
The Second Affiliated Hospital of Harbin Medical
University, Harbin, China
3
Department of Pharmacology, Harbin Medical
University, Harbin, China
1

Background and objectives: The present study has been
designed to investigate the preservation of lycopene for acute
myocardial infarction in vivo.
Methods: Wistar rats were randomly assigned either to
lycopene, acute myocardial infarction (AMI) model and the
normal groups. Lycopene group was given lycopene dissolved
in olive oil each day, while the model group and normal group
were given olive oil in the same amount. After 30 days, rats of
lycopene group and model group were performed left anterior
descending coronary artery ligation to establish AMI model.
The severity of AMI and parameters of oxidative stress were
assessed by the serum, histopathological exam and myocardial
homogenate.
Results: Pretreatment with lycopene significantly increase
the survival rate, reduce the infarct size and amount of myocardial enzymes. The alleviation of coagulative necrosis in ventricle of lycopene group was confirmed by the histopathological
exam. The comprehensive oxidative stress assessment indicated
lycopene significantly decreased the amount of the products of
lipid peroxidation, protein damage, and DNA damage, significantly increased the amount of antioxidative enzymes.
Conclusions: Lycopene had a myocardial preservation and
the effect might attenuate the intensified oxidative stress in

AMI and stimulate the generation of endogenous antioxidants.
Key words: Lycopene; Myocardial preservation; Acute
myocardial infarction; Oxidative stress; Rats

Key words: sweet cherries, polyphenolics, antioxidant activity

Abstracts

Ann Nutr Metab 2013;63(suppl 1):1–1960

1553


PO2665
ANTIOXIDATIVE
AND
HEPATOPROTECTIVE
EFFECTS OF GLOSSOGYNE TENUIFOLIA ON ACETAMINOPHEN-INDUCED LIVER DAMAGE IN MICE
Y. Lu1, Y. Tien1
1
Department of Nutritional Science, Fu-Jen Catholic
University, New Taipei City, Taiwan
Background and objectives: Acetaminophen (APAP), a
commonly used antipyretic and analgesic agent, can cause
acute liver failure by overdose. Glossogyne tenuifolia (GT),
Hsiang-Ju, used to prepare an herbal tea in Penghu, Taiwan,
has been shown in recent research to exhibit antioxidative,
antic-inflammatory, and anticarcinogenic properties. The present study investigates the antioxidative and hepatoprotective
effects of GT against acetaminophen-induced acute liver injury
in BALB/c mice.

Methods: The extracts of GT by various solvents (hot
water, 50% ethanol and 95% ethanol) were used to compare
the DPPH radical scavenging activity, reducing powder, total
phenolic contents, and trolox equivalent antioxidant capacity
(TEAC).
Results: The results showed that hot water extracts exert
an excellent antioxidative capacity and were thus used in the
animal experiment. The 30 male BALB/c mice were randomly
divided into negative control group (APAP 400mg/kg), positive control group (N-acetylcysteine 600 mg/kg bw) and two
GT groups at different dosages (100, 300 mg/kg respectively).
After the administration of different oral treatments for one
week, APAP was administered to the mice on the seventh day.
The results showed that mice treated with GT had significantly
decreased serum levels of alanine aminotrasferase (ALT), aspartate aminotrasferase (AST), and lipid TBARS (thiobarbituric acid-reactive substances), but total antioxidant capacity
(TEAC) was not affected. GT-H (300 mg/kg) increased catalase
activity in the liver; no prominent effects were demonstrated
on superoxide dismutase, glutathione S-transferase, glutathione peroxidase.
Conclusions: In conclusion, hot water extract of GT may
have potential antioxidant capacity and exert hepatoprotection
in mice.
Key words: acetaminophen, Glossogyne tenuifolia, hepatoprotective, antioxidative

PO2666
IN VITRO METABOLISM OF 3,5,6,7,8,3’,4’-HEPTAMETHOXYFLAVONE BY RAT LIVER MICROSOMES
N. Koga1, M. Matsuoka1, C. Ohta1, Y. Kato2, K. Haraguchi3, O. Kimura4, T. Endo4
1
Faculty of Nutritional Sciences, Nakamura Gakuen
University, Japan
2
Faculty of Pharmaceutical Sciences, Tokushima

Bunri University, Japan
3
Daiichi College of Pharmaceutical Sciences, Japan
4
Faculty of Pharmaceutical Sciences, Hokkaido
Health University, Japan
Background and objectives: 3,5,6,7,8,3’,4’-Heptamethoxyflavone (HepMF), a polymethoxyflavone present abundantly
in the citrus peel, have been reported to show biological properties such as the inhibition of P-glycoprotein and the suppression of growth of tumor cells. However, the mechanism
which the activities are exerted by HepMF or its metabolites
remains obscure. Therefore, the metabolism of HepMF was
investigated using rat liver microsomes and the effects of typical cytochrome P450 (P450) inducers, phenobarbital (PB),
3-methylcholanthrene (MC) and dexamethasone (DEX), were
also examined.
Methods: HepMF was incubated for 20 min at 37ºC with
rat liver microsomes, NADPH-generating system and HEPES
buffer (pH 7.4) under aerobic conditions. Rat liver microsomes were prepared from untreated, PB-treated, MC-treated
and DEX-treated rats. After incubation, an ice-cold methanol
was added and the incubation mixtures were centrifuged. The
supernatants were applied to HPLC.
Results: HepMF was metabolized to three major metabolites (M-2, M-3 and M-4) and four minor metabolites (M-1,
M-5, M-6 and M-7) by liver microsomes of untreated rats
and the amount of M-2, M-3 and M-4 was 0.42, 0.56 and 0.12
nmol/min/mg protein, respectively. PB treatment increased
three major metabolites to about 2-fold and M-1 to 4.4-fold of
untreated rats significantly. MC treatment increased both M-2
and M-4 to about 3-fold of untreated rats and also accelerated
the formation of M-5, M-6 and M-7 to 244-, 16- and 5-fold of
untreated rats, respectively. Moreover, DEX showed the similar induction pattern to PB. From the spectral data of MS and
1H-NMR, three major metabolites, M-2, M-3 and M-4, were
assumed to be 4’-demethylated (OH), 7-OH and 6-OH metabolites, respectively. The chemical structures of M-1, M-5, M-6

and M-7 remain unclear because of their low yields.
Conclusion: These results suggest that CYP2B, CYP1A and
CYP3A enzymes are responsible for HepMF metabolism.
Key words: polymethoxyflavone, metabolism, rat, liver

1554

Ann Nutr Metab 2013;63(suppl 1):1–1960

Abstracts


PO2667
PLANT-BASED FOODS AS A SOURCE OF LIPOTROPES FOR PREVENTING HEPATIC STEATOSIS
Fardet1, J. Martin1, J. Chardigny1
1
Inra, CLERMONT-FERRAND, France
Background and objectives: Lipotropes are food components that limit excessive hepatic triglyceride contents or
steatosis. Steatosis is often associated with obesity and type
2 diabetes, and may lead to more serious pathologies such as
steatohepatitis, hepatic fibrosis and cirrhosis, or cancer. Yet,
whereas steatosis concerns several millions people worldwide,
the lipotropic potential of foods has never been studied; and
lipotrope-rich foods remain quite unknown. Our objective was
to characterize and quantify the lipotropic potential of plantbased foods.
Methods: From lipotrope contents found in literature and
nutritional tables, 132 plant-based foods and 8 lipotropes (betaine, choline, myo-inositol, methionine, niacin, pantothenic
acid, folates and magnesium) could have been selected. Plantbased food lipotropic potential was expressed into a new index,
the Lipotropic Capacity (LC) that integrates the sum of the 8
lipotropic densities relative to a reference food.

Results: Vegetables are the best source of lipotropes on a
100 kcal-basis and plant-based foods are a more diversified
source - but complementary - of lipotropes than animal-based
products. Technological processes reduce plant-based foods lipotropic potential by around 20%: while refining is the most
drastic treatment, fermentations may tend to increase lipotrope densities. By comparing lipotrope consumption via both
French standard diet (INCA 2 survey) and Food guide pyramid, we evaluated that our consumption in betaine, choline
and myo-inositol may be increased: this can be easily reached
by choosing lipotrope-dense foods like beetroot, spinash or coffee.On a one euro-basis, grains products (i.e. cereals, nuts and
leguminous and oleaginous seeds) are the best compromise
between a high LC and a cheap supply in lipotropes.
Conclusions: The LC allows easily ranking foods according
to their lipotropic potential. But, it remains indispensable to
carry out studies in humans to relate food LC with prevalence
of hepatic steatosis.
Key words: Lipotropes, plant-based foods, lipotropic capacity, technological processes, consumption

PO2668
NOURISHMENT FUNCTION IMPROVEMENT OF
THE WHEAT PRODUCTS BY THE ULVA PERTUSA
ADDITION
A. Yamamoto1, M. Moriyama2, A. Koide3, K. Matsumoto4, . Mamiya3, T. Yamauchi3
1
Department of Food Science and Nutrition,Aichi
Gakusen College, Japan
2
Faculty of Human Life and Environmental
Sciences,Aichi Gakusen University, Japan
3
Faculty of Human Life and Environmental
Sciences,Nagoya Women’s University, Japan

4
Department of Applied Life Studies,College of
Nagoya Women’s University, Japan
Background and objectives: In late years, eating habits
were influenced by westernized countries. Lack of a mineral
and dietary fiber in meal was related to diabetes mellitus. The
meal intake standard of magnesium (Mg) is 370 mg/day, but
there are few intakes with 250 mg. We processed Ulva pertusa (Ulva) which is unused food resources into wheat products.
The chlorophyll content of Ulva is high and addition of it which
is green algae in familiar food served as a source of Mg supply.
Noodles and bread were prepared with addition of Ulva. Examination of chlorophyll and Mg were done to assess cooking
loss and its effectiveness.
Methods: After hanging Ulva to a lyophilizer, Ulva powder was sieved using280 microns. Ulva noodles were variably
added with Ulva powder with a density of 0%, 3%, 5%, 8% and
13% while Ulva bread addition of Ulva powder were 0%, 1%,
2%, and 3% respectively. Ingredient analysis for the quantity of
Mg and chlorophyll were performed by Arnon Method and Ion
Chromatograph Method.
Results: The chlorophyll content correspondingly increased
with Ulva density addition. The quantity of Mg in 13% Ulva
powder noodles increased to 1,030mg per 100 g. After boiling,
Mg quantity was 530 mg. In bread, Mg quantity was 400 mg per
100 g of 3% of Ulva powder bread before bake. An amount of
151 mg was obtained after baking.Quantity of Mg in noodles
and bread with increasing addition of Ulva density was similar
in quantity increase of chlorophyll.
Conclusions: Increasing Ulva powder density of all wheat
products put up a nourishment ingredient than additive-free
noodles. It was suggested that it could become a source of Mg
supply by the addition of Ulva.

Key words: Ulva pertusa, Magnesium, Chlorophyll, Wheat
products

Abstracts

Ann Nutr Metab 2013;63(suppl 1):1–1960

1555


PO2669
DETERMINATION OF PHENOLIC ACIDS IN INDIGENOUS LEAFY VEGETABLES OF BOTSWANA BY
HIGH PERFORMANCE LIQUID CHROMATOGRAPH
S T P. Matenge1, N. Dlamini2, K. Mathiba2
1
National Food Technology Research Centre, Kanye,
Botswana
2
CSIR, BioSciences Unit (Natural Products and Agro
processing Platform), Pretoria, South Africa
Background and Objectives: Plant materials especially
green leafy vegetables contain phytochemicals whose consumption has been associated with protecting the human body
from chronic diseases.
Methods:With the aim to promote the utilisation of indigenous leafy vegetables, the present study separated, identified and quantified using HPLC-DAD the main phenolic
compounds in indigenous leafy vegetables [Amaranth (Amarunthus spp.), Spider plant (Cleome gynandra), Cowpea leaves
(Vignaunguiculate) and Black nightshade (Solanaceae spp)]of
Botswana.
Results: 3,4-dihydroxbenzoic acid (7.56 ± 0.61mg/100g),
chlorogenic acid (12.34 ± 0.95mg/100g) and ferulic acid (3.63
± 0.09mg/g) in Amaranth spp., 3,4-dihydroxbenzoic acid (7.25

± 0.11mg/100g), 4-hydroxbenzoic acid (3.85 ± 0.14mg/100g),
p-coumaric acid (1.60 ± 0.25mg/100g) and ferulic acid (4.75
± 0.25mg/100g) where found in Vignaunguilate, vanillic acid
(1.13 ± 0.09mg/100g), chlorogenic acid (31.32 ± 4.42mg/100g)
and ferulic acid (2.38 ± 0.09mg/100g) were found in Cleome
gynandra, 3,4-dihydroxbenzoic acid (6.68 ± 0.05mg/100g) and
ferrulic acid (7.93 ± 0.94mg/100g) were found in Solanaceae
spp. Ferulic acid (2.38 ± 0.09mg/100 – 7.93 ± 0.94mg/g) appeared in all the samples analysed, however, chlorogenic acid
(12.34 ± 0.95mg/100g – 31.32 ± 4.42mg/100g) was the most
abundant.
Conclusion: Vegetables investigated in this study are shown
to be rich sources of polyphenols which contribute to their
health promoting properties. Therefore the use of these vegetables as a source of polyphenols could offer enormous opportunities for the functional food industry. In addition, it is critical
to create awareness regarding diet related health benefits of
these vegetables.This is the first report on identification of phenolic acids in the extracts of four indigenous leafy vegetables
grown in Botswana.
Key words: Polyphenols, Amarunthus sp., Cleome gynandra, Vignaangulate, Solanaceae (nightshade), HPLC

1556

Ann Nutr Metab 2013;63(suppl 1):1–1960

PO2670
INFLUENCE OF PINE BARK EXTRACT POLYPHENOLS ON INTESTINAL MICROFLORA
T. Sanchez Moya1, R. López Nicolás1, D. Planes Muñoz1,
P. Peso Echarri1, C. González Bermúdez1, V. Gómez Gómez1, A. López Fernandez1, L. Fernández Palacios1, C.
Frontela Saseta1, C A. Martínez Graciá1
1
Department of Food Science and Nutrition, Faculty
of Veterinary Sciences of University of Murcia, Murcia, Spain

Background and objectives: Phenolic compounds exert
antioxidant properties and beneficial effects on human health,
being abundant in fruits and other vegetables, as cereals and
legumes. As a concentrated of water-soluble polyphenols, Pine
(Pinus pinaster Ait) bark extract (PBE) has been demonstrated
to have strong antioxidant properties. In this study, the antimicrobial activity of caffeic acid, gallic acid, ferulic acid, chlorogenic acid and taxifolin; and PBE, against Escherichia coli
O157:H7, Enterobacter sakazakii, Listeria monocytogenes and
Staphylococcus aureus, Escherichia coli, Enterococcus faecalis,
and the effect on Lactobacillus casei rhamnosus and Lactobacillus gasseri, was determined.
Methods: Taxifolin, ferulic, gallic, chlorogenic and caffeic
acids, and PBE were analysed by HPLC. Each phenolic compound (1.25, 0.6, 0.3, 0.15, 0.07, 0.04 and 0.02 mg/mL) was tested against different bacterial strains to determine their effects
on bacterial growth, using the microtiter plate method.
Results: Increased concentration of phenolics led to an inhibition of bacterial growth with differences depending of the
compound. The growth of S. aureus and Lactobacillus gasseri
was inhibited at 0.15 at 1.25 mg/mL and 0.3-0.6 mg/ml of caffeic acid, respectively. Meanwhile L. casei rhamnosus was not
affected by this compound. Regarding gallic acid, the inhibitory effect was highly significant for S. aureus and E. faecalis, at
concentrations of 0.02-1.25 and 0.04-1.25 mg/mL, respectively.
Taxifolin showed antilisterial effect at concentrations of 1.25
mg/mL with growth inhibition values of 97%. The inhibitory
effect of PBE was highly significant on the growth of E. coli
O157:H7Nevertheless, the growth probiotic bacteria growth
was poorly affected by most of tested phenolic compounds.
Conclusion: PBE and its constituents showed selective antimicrobial activity on intestinal bacteria. Moreover, we have
demonstrated the protective effect of polyphenoles and PBE on
probiotic bacteria as L. casei rhamnosus and L. gasseri.
Key words: pine bark extract, polyphenols, intestinal microflora

Abstracts



PO2671
SYNERGISTIC ANTIOXIDANT ACTIVITY OF PROTEIN HYDROLYSATES FROM DIFFERENT SOURCES OBTAINED BY ENZYMATIC HYDROLYSIS
R. Castro1, H. Sato1
1
Department of Food Science, School of Food Engineering, University of Campinas, Campinas, São
Paulo, Brazil
Background and objectives: Soy protein isolate (SPI), bovine whey protein (BWP) and egg white protein (EWP) hydrolysates were prepared using different microbial proteases
to produce peptides with antioxidant activity (DPPH radical
scavenging).
Methods: The process parameters: enzyme type, substrate
and enzyme concentrations, and hydrolysis time were further
investigated for each protein source. After defining the most
appropriate process parameters for enzymatic hydrolysis, the
interactions amongst the hydrolysates with maximal antioxidant activity obtained from the different sources, were studied
using a simplex centroid mixture design.
Results: SPI hydrolysates with maximal antioxidant activity
were obtained with a substrate concentration of 90 mg.mL-1
and 70.0 U of a commercial protease from Aspergillus oryzae
per mL of reaction mixture (U.mL-1), with a hydrolysis time
between 120 and 180 min. For BWP hydrolysates the hydrolysis parameters were: substrate concentration of 8.0% plus 70.0
U.mL-1 of a protease from Aspergillus oryzae produced by solid-state fermentation, and a hydrolysis time in the range from
60 to 240 min. The hydrolysates prepared from 3.0% EWP plus
20.0 U.mL-1 of a commercial protease from Aspergillus oryzae,
obtained after 120 minutes of hydrolysis time, presented high
DPPH radical scavenging. In the antioxidant study of seven
formulations containing SPI, BWP and EWP hydrolysates, alone or in combinations, a synergetic effect was found for the
ternary mixture. In a mixture composed of SPI (1/3), BWP
(1/3) and EWP (1/3), increases of 184.1, 164.7 and 217.5% in
DPPH radical scavenging were observed as compared to the
respective isolated hydrolysates. However the binary mixture

BWP (1/2) plus EWP (1/2), showed an antagonistic effect. The
interactions between the binary mixtures: SPI (1/2) plus BWP
(1/2) and SPI (1/2) plus EWP (1/2) were not statistically significant (p < 0.05).
Conclusions: The results suggest that the enzymatic hydrolysis of proteins from different sources and their mixtures can
improve the antioxidant activities.
Key words: protease, protein hydrolysates, antioxidant activity, synergistic effect

Abstracts

PO2672
FUNCTIONAL PROPERTIES AND INHIBITION OF
THE RELATIVE LIPID ACCUMULATION OF PROTEIN HYDROLYSATES OBTAINED BY ENZYMATIC
HYDROLYSIS USING STATISTICAL MIXTURE DESIGN
R. Castro1, R. Inacio2, A. Oliveira2, H. Sato1
1
Department of Food Science, School of Food Engineering, University of Campinas, Campinas, São
Paulo, Brazil
2
Department of Anatomy, Institute of Biology, University of Campinas, São Paulo, Brazil
Background and objectives: Statistical design mixtures of
soy protein isolate (SPI), bovine whey protein (BWP) and egg
white protein (EWP) were used to study the effects of the different protein sources and their mixtures on the functional properties (solubility, heat stability, emulsion activity index and
foaming capacity) and inhibition of the relative lipid accumulation (RLA) in 3T3-L1 preadipocytes during differentiation.
Methods: The hydrolysates were prepared in Erlenmeyers
flasks containing 50 mL of protein solution (100 mg.mL-1) and
a commercial protease from Aspergillus oryzae. The solution
was incubated for 2h under the optimum pH and temperature
conditions for the enzyme (pH 5.0 at 50.0°C).
Results: For functional properties, the enzymatic hydrolysis increased the solubility of the proteins, excepted for EWP
and the mixtures containing it. However, the hydrolysates exhibited a tendency to decrease the foaming capacity and heat

stability. A synergetic effect was found for the emulsion activity index with two binary formulations and the ternary mixture, with increases of up to 12-fold. The EWP hydrolysates
showed the highest RLA suppression (19.7%) followed by BWP
(16.4%) and SPI (8.7%). Two binary mixtures and the ternary
formulation showed similar RLA suppression compared to the
isolated protein sources. The hydrolysates obtained using equal
proportions of SPI and BWP, and of BWP and EWP, showed
RLA suppressions of 11.5 and 12.8%, respectively, while the
ternary mixture containing SPI (1/3), BWP (1/3) and EWP
(1/3) showed an RLA suppression of 14.5%.
Conclusions: The results suggest that the enzymatic hydrolysis of proteins from different sources and their mixtures can
improve some functional properties and increase the suppression of RLA in 3T3-L1 cell differentiation.
Key words: protein hydrolysis, mixture design, functional
properties, relative lipid accumulation, 3T3-L1.

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PO2673
IN VITRO METABOLISM OF DIOSMETIN AND HESPERETIN BY RAT LIVER MICROSOMES
C. Ohta1, Y. Kato2, K. Haraguchi3, N. Koga1
1
Faculty of Nutritional Sciences, Nakamura Gakuen
University, Japan
2
Faculty of Pharmaceutical Sciences, Tokushima
Bunri University, Japan
3
Daiichi College of Pharmaceutical Sciences, Japan

Background and objectives: Flavonoids in fruits and vegetables are transformed to hydroxy-metabolites via hydroxylation and demethylation in animals. To elucidate the relationship between their chemical structures and metabolic patterns,
the metabolism of a flavone (diosmetin) and a flavanone (hesperetin) by rat liver microsomes was compared and the effects
of inducers and inhibitors of cytochrome P450 (P450) on their
metabolisms were examined.
Methods: Rat liver microsomes were prepared from untreated, phenobarbital (PB)-treated and 3-methylcholanthrene
(MC)-treated rats. Each flavonoid was incubated for 20 min
at 37°C with rat liver microsomes, NADPH and HEPES buffer
(pH 7.4) under aerobic conditions. After addition of methanol,
the incubation mixtures were centrifuged to remove the denatured protein. For analyses of the metabolites, the supernatants
were applied to HPLC. α-Naphthoflavone (αNF), furafylline
(FFL) and ketoconazole (KCZ) were used as P450 inhibitors
for CYP1A1, CYP1A2 and CYP3A1, respectively.
Results: In untreated microsomes, diosmetin and hesperetin were mainly oxidized to give 4’-demethylated metabolites,
luteolin and eriodictyol, with the rates of 0.47 and 0.16 nmol/
min/mg protein, respectively. In hesperetin metabolism, three
minor metabolites were also formed. PB-treatment showed
a slight increase (about 1.5-fold of untreated rats) of both luteolin and eriodictyol, whereas MC-treatment increased both
metabolites remarkably to 5.9-fold and 10.9-fold of untreated
rats, respectively. However, the amount of eriodictyol formed
were 30-64% of that of luteolin formed in all cases. 0.5 mM
αNF inhibited the formation of luteolin and eriodictyol to 45%
and 18% of control, respectively. 0.25 mM KCZ inhibited both
metabolites to 30% of control.
Conclusion: These results suggested that CYP1A and
CYP3A enzymes are the most important P450 isoforms in the
metabolism of both flavonoids in rat and that diosmetin having the flavone skeleton is a better substrate for CYP1A and
CYP3A enzymes than hesperetin having flavanone skeleton.
Key words: flavone, flavanone, metabolism, rat, liver

PO2674

NOVEL ANXIOLYTIC-LIKE PEPTIDE RELEASED
FROM ALPHA-S-CASEIN BY GASTROINTESTINAL
PROTEASES
T. Mizushige1,2, Y. Sawashi1, A. Yamada1, R. Kanamoto1,
K. Ohinata1
1
Divisionof Food Science and Biotechnology,
Graduate School of Agriculture, Kyoto University,
Kyoto, Japan
2
Research Unit for Physiological Chemistry, C-PIER,
Kyoto University, Kyoto, Japan
Background and objectives: Our previous findings demonstrated that dipeptide YL exhibited orally active anxiolytic-like activity comparable to diazepam [1]. The YL sequence is often observed in the primary structure of natural food
proteins. We then investigated whether YL analogues are released from bovine alpha-s-casein by gastrointestinal proteases.
Methods: YL analogues in the pepsin-pancreatin digestion
of alpha-s-casein were quantified by LC-MS. Synthetic peptides
were prepared by the F-moc strategy. We investigated anxietylike behavior using two behavioral tests, the elevated plus-maze
and the open-field, in mice. Peptide or digest dissolved in saline was administered intraperitoneally and/or orally 30 min before the test. Antagonists of neurotransmitters associated with
anxiolytic-like activity were co-administered intraperitoneally
with peptide 30 min before the test. A percentage of the entry
time in open arms or in the center circle was measured.
Results: We found that YLG, corresponding to alpha-s1casein(91-93), was more effectively released from alpha-s-casein than YL by pepsin-pancreatin digestion, mimicking gastrointestinal enzymatic conditions. Using the synthetic model
peptide, we determined that trypsin cleaved the N-terminus of
YLG, and elastase and carboxypeptidase contributed to cleavage of the C-terminus of that. YLG as well as YL exhibited
potent anxiolytic-like activity after oral administration. The
anxiolytic-like activity of YLG was inhibited by WAY100135,
SCH23390 or bicuculline, an antagonist of serotonin 5-HT1A,
dopamine D1 and GABAA receptors, respectively; however,
YLG has no affinity for these receptors. These results suggest
that YLG exhibits anxiolytic-like activity via the releases of

serotonin, dopamine, and GABA, which are a novel neuronal
pathway common to YL. The pepsin-pancreatin digestion of
alpha-s-casein also exhibited anxiolytic-like activity.
Conclusions: We found that YLG, a novel anxiolytic-like
peptide, released from alpha-s-casein after pepsin-pancreatic
digestion mimicking gastrointestinal conditions, exhibits
anxiolytic-like activity in mice.[1] Kanegawa N et al. (2010)
FEBS Lett 584:599-604.
Key words: emotional behavior, anxiolytic, peptide, bovine
milk protein

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Ann Nutr Metab 2013;63(suppl 1):1–1960

Abstracts


PO2675
PRODUCTION OF OIL PALM (ELAEIS GUINEENSIS)
KERNEL PROTEIN HYDROLYSATES WITH ANTIOXIDANT PROPERTIES AND APOLIPOPROTEIN B
SECRETION INHIBITION
S K. Chang1, H. Hamajima2, I. Amin1,5, T. Yanagita2,3, M
E. Norhaizan1, B. Mohamad Taufik Hidayat4
1
Department of Nutrition and Dietetics, Faculty
of Medicine and Health Sciences, Universiti Putra
Malaysia, Selangor, Malaysia
2
Jofuku Frontier Laboratory, Saga University, Japan

3
Department of Health and Nutrition Sciences,
Faculty of Health and Human Welfare Sciences,
Nishikyushu University, Kanzaki, Saga, Japan
4
Department of Human Anatomy, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia,
Selangor, Malaysia
5
Laboratory of Halal Science Research, Halal Products Research Institute, Universiti Putra Malaysia,
Serdang, Selangor, Malaysia
Background and objectives: Plant proteins played essential roles in human nutrition. Palm oil was the most important
oilseed crops globally. There was 12-16% of crude protein in
oil palm kernel. However, little was known about the functional properties of the kernel protein. Hence, this study aimed
to produce the protein hydrolysates and characterize its health
promoting properties.
Methods: The protein isolate used for the generation of
protein hydrolysates was obtained by alkaline extraction and
acid precipitation. Then, oil palm kernel protein hydrolysates
(OPKH) were produced by protease and pepsin-pancreatin hydrolysis. SDS-PAGE was used for qualitative assessment of its
polypeptide profiles. Amid acid profile was analyzed by Pico
Tag Amino Acid Analyzer. The effect of both protein hydrolysates (PH) on apoB secretion and lipid metabolism in vitro
was undertaken using HepG2 cells in ApoB Microwell ELISA
Assay Kit (AlerCHEK). In vitro antioxidant activity (AA) was
determined using ferric reducing antioxidant power (FRAP)
and 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)
(ABTS) assay.
Results: The estimated polypeptide molecular mass of both
protein hydrolysates from reducing SDS-PAGE were 7, 8, 11
and 12 kDa while only one protein band was observed from
non-reducing SDS-PAGE, 50 kDa. Both protein hydrolysates

were richest in Aspartic acid and Glutamic acid (> 35% of total
amino acids), followed by sulfur amino acids, lysine and valine. Both PH (1 mg/ml) reduced apoB secretion and its triglycerides and cholesterol synthesis significantly. Protease and
pepsin-pancreatin hydrolysates reduced apoB secretion 45.4%
and 32.6% respectively compared to control. OPKH (5 mg/ml)
Abstracts

were highly effective in scavenging ABTS radicals and with
high reducing power (FRAP) compared to many other seed
PH. There was a dose-dependent relationship between both
PH and AA.
Conclusions: OPKH have good health promoting properties as a source of nutraceuticals contributed by its lipid lowering and antioxidative properties.
Key words: Oil palm kernel; protein hydrolysates; apoB secretion; antioxidant activity

PO2676
DEVELOPMENT OF A REPROTER ASSAY SYSTEM
TO SCREEN FOR CHEMICALS MIMICKING THE
ANTI-AGING EFFECTS OF CALORIE RESTRICTION
T. Chiba1, T. Tsuchiya2, I. Shimokawa2,
1
Biomedical Gerontology Laboratory, Faculty of
Human Sciences, Waseda University, Tokorozawa,
Japan
2
Department of Investigative Pathology, Unit of Basic Medical Science, Graduate School of Biomedical
Sciences, Nagasaki University, Nagasaki, Japan
Background and objectives: Caloric restriction (CR) in
normal mice, and suppression of the growth hormone (GH)
/ insulin-like growth factor-I (IGF-I) pathway in spontaneous
mutant Ames dwarf (DF) mice extends their lifespan and delays the onset of age-dependent disorders. In combination, these two lifespan-extending interventions have an additive effect
upon DF mice. In this study, we tried to identity the intracellular signaling pathways of CR beneficial effects and develop a

system to screen anti-aging chemicals.
Methods: We previously identified dozens of genes up-regulated in the liver of DF and CR mice by using DNA microarray. Motif analysis was performed to identify consensus sequence motifs in the upstream of these putative pro-longevity
genes. We tried to identify binding factors of these motifs using
gel shift assay. We also constructed the reporter assay system to
screen CR mimetics generating the constitutive cell lines and
the transgenic mice harboring the motif sequence.
Results: We found that one of the synthesized sequences of
identified motifs bound to hepatocyte nuclear factor-4 (HNF4) alpha, an important transcription factor involved in glucose
and lipid metabolism. When the reporter construct, containing
an element upstream of a secreted alkaline phosphatase (SEAP)
gene, was co-transfected with HNF-4 and its regulator peroxisome proliferator-activated receptor-gamma coactivator 1 (PGC1) alpha, activity of SEAP was increased dose-dependently in
comparison to untransfected controls. Moreover, transgenic
mice established using this construct showed increased SEAP
activity both CR and resveratrol-fed mice in comparison to ad
Ann Nutr Metab 2013;63(suppl 1):1–1960

1559


libitum fed mice. Furthermore, oxidative stress resistance was
correlated with SEAP activity of these transgenic mice.
Conclusions: These data suggest that our bioassay would be
useful to screen for CR mimetic photochemical both in vitro
and in vivo.
Key words: Aging, calorie restriction, photochemical,
HNF-4, PGC-1

Conclusions:The content of bioactive compounds is affected more by expiration date rather than by heat treatment.It
is also interesting to indicate that transfer of bioactive compounds between vegetables during cooking can be observed.
AcknowledgmentsThe authors thank Andalusian Regional

Government (Regional Ministry of Innovation, Science and
Enterprise) and FEDER for financial support Project Ref.P11AGR-7034.
Key words: bioactive compounds, UHPLC-MSMS, readyto-heat vegetables.

PO2677
MONITORING OF BIOACTIVE COMPOUNDS IN
READY-TO-HEAT VEGETABLES: EVALUATION OF
CONTENT, STABILITY AND COMPARISON
M I. Alarcon-Flores1, F. Hernández-Sánchez1, R. Romero-González1, P. Plaza-Bolaños1,2, A. Garrido-Frenich1
1
Research Group Analytical Chemistry of Contaminants, Department of Chemistry and Physics,
Research Centre for Agricultural and Food Biotechnology (BITAL), University of Almería, Agrifood
Campus of International Excellence, Almeria, Spain
2
Department of Analytical Chemistry, University of
Granada, Granada, Spain
Background and objectives: Bioactive compounds are present in fruit and vegetables and they are recognized for their
benefits for health. Because the advantages of ready-to-heat
vegetables, as time saving, healthy characteristics and low-cost,
this market is currently the fastest growing subsector of food
industry.The purpose of this work is the study and comparison
of the content of several families of bioactive compounds in
ready-to-heat vegetables, as pepper, tomato, carrot, cauliflower
and broccoli, before and after applying a microwave heating.
In a second step, the evolution of the content of bioactive compounds during shelf life of these products has been evaluated.
Methods:The extraction procedure carried out was based
on a solid liquid extraction using methanol:water (80:20,v/v).
The extract was injected into the ultra-high-performancechromatogram coupled to triple quadrupole mass analyzer.
Results:The total content of bioactive compounds in pepper was slightly increased after heat treatment. The content of
bioactives was higher when the sample was heated after expiration date.A bag of a mixture of broccoli, cauliflower and carrot

was evaluated, although each matrix was always analyzed separately. Caffeic acid appeared after heat treatment, and it must
be considered that it was a degradation product of chlorogenic
acid. The content of total bioactive compounds in carrots increased after heating. In general, it was observed that expiration date affects more than heat treatment to the total content
of bioactives. Nonetheless, differences were not statistically significant in most cases, except for broccoli.
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Ann Nutr Metab 2013;63(suppl 1):1–1960

PO2678
A COMPARISON OF BIOACTIVE COMPOUNDS IN
FRESH AND FRESH CUT VEGETABLES
M I. Alarcon-Flores1, R. Romero-González1, J L. Martinez-Vidal1, F J. Egea-Gonzalez1, A. Garrido-Frenich1
1
Group “Analytical Chemistry of Contaminants”,
Department of Chemistry and Physics, Research
Centre for Agricultural and Food Biotechnology
(BITAL), University of Almería, Agrifood Campus of
International Excellence, Almería, Spain
Background and objectives: Consumption of fruit and vegetables could help to prevent diseases. These beneficial effects
have been attributed to bioactive compounds.Nowadays,there
is an increased interest of the consumers by fresh-cut products.
Therefore,it is important to investigate if the minimal processing may provoke losses of important bioactive compounds.
The purpose of this work was the comparison of the content
of several families of bioactive compounds in fresh and freshcut products, as tomato, eggplant, grape, carrot and broccoli, in
order to get a comprehensive view of the presence of bioactive
compounds in fresh and fresh-cut products. Other variables
such as type of cut and shelf life were also evaluated.
Methods:The extraction procedure was based on a simple
solid-liquid extraction with methanol:water (80:20,v/v), and
the extract was injected into an ultra-high-performance-chromatographic system coupled to triple quadrupole mass analyser.

Results:Phenolic acids were the most abundant class of
phytochemicals in tomato, carrot and eggplant, whereas glucosinolates were majority in broccoli and flavonols in grape.
When bioactive compounds contents were compared in fresh
and fresh-cut products, the concentrations in fresh products
were higher than in fresh-cut, except for phenolic acids in eggplant, although in most cases the difference was not statistically
significant. On the other hand,it was observed that phenolic
acids were the class most affected by type of cut.Furthermore, it
was noted that the storage at lower temperature (4 ºC) provides
higher levels of the assayed compounds.
Abstracts


Conclusions:Fresh and fresh-cut products have similar
beneficial properties with respect to their content in bioactive
compounds, except tomato, which contains a higher amount of
phenolic acids in fresh products.
Acknowledgments: The authors are grateful to Andalusian Regional Government (Regional Ministry of Innovation,
Science and Enterprise) and FEDER for financial support Project Ref.P11-AGR-7034 and to Guzmán-Gastronomía.
Key words: fresh vegetables, fresh-cut vegetables, bioactive
compounds,UHPLC-MSMS

PO2680
FUNCTIONAL/NUTRACEUTICAL EVALUATION OF
CITRUS PEEL AGAINST DYSLIPIDEMIA
M. Butt1, H. Ashraf1, F. Anjum1
1
National Institute of Food Science and Technology,
University of Agriculture, Faisalabad, Pakistan
Background and objectives: Functional/nutraceutical
foods are the core element of diet based therapy owing to

their health enhancing potential beyond the basic function of
supplying nutrients. Citrus peel, a byproduct of food processing industry has group of bioactive components that play a
significant role against various physiological threats. The most
prevalent polymethoxylated flavones are tangeretin and nobiletin that have physiological importance with enormous food
and pharmaceutical applications. The present project was designed to establish the dyslipidemic potential of citrus peel extract with special reference to their active ingredients.
Methods: The citrus peel was characterized followed by product development phase and nutraceutical evaluation for the
management of dyslipidemia through experimental rats modeling. The citrus peel extracts were prepared by using methanol,
ethanol and water and checked for FRAP, DPPH, flavonoids,
flavonols and ABTS. Functional/nutraceutical cakes and biscuits were also prepared. Further, the citrus peel extract and
powder were assessed for cholesterol, low density lipoprotein
(LDL), high density lipoprotein (HDL) and triglycerides (TG)
in Sprague Dawley rats.
Results: The citrus peel methanolic extract showed higher
total polyphenols, FRAP, DPPH, flavonoids, flavonols & ABTS
values in comparison with other extracts. Functional/nutraceutical cakes and biscuits were prepared after adding citrus peel
extract and powder. Bioevaluation comprised of three groups
of rats feeding on control diet, diet containing 10% citrus peel
and diet containing 5% citrus extract. Cholesterol and LDL
were significantly reduced ranging from 6.01 to 7.25 and 8.78
and 11.39%, respectively with subsequent rise in HDL. Furthermore, the values for liver and kidney functions tests were
within the normal range showing the safety prepared products.
Abstracts

Conclusions: In the nutshell, citrus peel based edibles have
potential to curtail different physiological malfunctioning thus
should be encouraged in diet based therapy among the vulnerable segments.
Key words: Citrus peel, functional/nutraceutical, dyslipidemia, cholesterol, HDL

PO2681
FAVORABLE EFFECT OF CONSUMPTION OF LILY

BULB ON COLONIC LUMINAL ENVIRONMENT IN
RATS FED A HIGH-FAT DIET
Y. Okazaki1, E. Fukuda1, H. Chiji1, N. Kato2
1
Faculty of Human Life Sciences, Fuji Women’s University, Ishikari, Japan
2
Graduate School of Biosphere Science, Hiroshima
University, Higashi-Hiroshima, Japan
Background and objectives: The bulbs of lily (Liliumleichtliniivar. maximowiczii) are traditionally consumed in East
Asia, including Japan, as both food and oriental medicine. It
contains high amounts of starch, protein, and dietary fibers
including glucomannan. This study investigated the effect of
dietary lily bulb on the colonic luminal environment in rats fed
a high-fat (HF) diet.
Methods: In experiment 1, male SD rats were fed a HF diet
(30% beef tallow)with or without 5% or 10% raw lily bulb powder for 3 weeks. In experiment 2, rats were fed a HF diet with
or without 7% steamed or raw lily bulb powder for 3 weeks.
Intestinal luminal variables, including IgA, mucins (indices of
intestinal immune and barrier functions, respectively), organic
acids and microflora were measured.
Results: In experiment 1, Food intake and growthwere
reduced in the 10% raw lily bulb group. Dietary raw lily bulb
significantly increased fecal IgA and mucinsand cecalLactobacillales in a dose dependent manner. In experiment 2, Food
intake and growth were unaffected by the dietary treatment.
Fecal IgA and cecalLactobacillales were significantly elevated
in the raw lily bulb group, and similar trends were observed
in the steamed lily bulb group. Consumption of both steamed
and raw lily bulb markedly elevated fecal mucins, the weight of
cecaldigesta and cecal organic acids, including lactate, propionate, acetate and butyrate. Compared with the control group,
fecal ?-glucosidase activity was significantly higher in the raw

lily bulb group, but not in the steamed lily bulb group. Dietary
steamed lily bulb significantly reduced mesenteric adipose tissue weight and increased fecal triglyceride excretion.
Conclusion:The results suggest afavorable effect of lily bulbon colon health in rats fed a HF diet.
Key words: lily bulb, colonic luminal environment, rats
Ann Nutr Metab 2013;63(suppl 1):1–1960

1561


PO2682
THREE NOVEL PROBIOTIC STRAINS ISOLATED
FROM FECES OF BREAST-FED INFANTS MODULATE THE IMMUNE SYSTEM OF HEALTHY ADULTS
J. Plaza-Diaz1,2, C. Gomez-Llorente1,2, L. Campaña-Martin1,2, M. Bermudez-Brito1,2, E. Matencio3, F. Romero3, A.
Perez de la Cruz4, A. Gil1,2, L. Fontana1,2, other members
of the PROBIENSA group
1
Dept. Biochemistry & Molecular Biology II, School
of Pharmacy, Campus de Cartuja, Granada, Spain
2
Institute of Nutrition & Food Technology “Jose Mataix”, Biomedical Research Center, Granada, Spain
3
Hero Global Technology Center, Hero Spain, S.A.,
Alcantarilla, Murcia, Spain
4
Unit of Nutrition and Dietetics, Virgen de las Nieves
Hospital, Granada, Spain
Background and objectives: Probiotics are microorganisms that provide health benefits to the host when ingested
in adequate amounts. We have described the identification and
characterization of 3 novel bacterial strains (Lactobacillus paracasei CNCM I-4034, Bifidobacterium breve CNCM I-4035
and Lactobacillus rhamnosus CNCM I-4036), which were

isolated from the feces of exclusively breast-fed infants. Prior
to its evaluation for beneficial effects in humans, safety of any
probiotic strain must be demonstrated. Here we describe a trial
with healthy volunteers to test the safety of the aforementioned
strains.
Methods: One hundred and three healthy patients underwent a 15-day washout period after which they were randomly and blindly divided to receive a placebo or a daily capsule containing 10E10 CFUs one of the 3 strains for 30 days.
The intervention period was followed by a second washout of
another 15 days. Patients did not take any fermented product
for the entire duration of the study.
Results: Three volunteers dropped out of the study. No patient had side effects of any kind. Volunteers that received the
placebo showed a decrease in serum IL-12 and IL-4 concentrations, as well as decreased populations of Atopobium, Lactobacillus and Bifidobacterium in feces. IgA concentration and
Clostridium coccoides population increased in the feces, whereas IL-12 decreased in serum, of patients fed B. breve CNCM
I-4035Intestinal colonization occurred in 86% of the patients
fed L. rhamnosus CNCM I-4036In these patients, increases in
the population of Clostridium leptum in feces, and IL-10, IL4, IL-10/IL-12 in serum, together with decreases in IL-12 and
TNF-alpha/lL-10, occurred. Patients fed L. paracasei CNCM
I-4034 showed augmented IL-10/IL-12 ratio.
Conclusions: Intake of the three novel probiotic strains by
healthy patients is safe. Altogether, results obtained warrant the
undertaking of similar studies in patients affected of pathologies such as intestinal-related diseases.
Key words: probiotics, safety.
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Ann Nutr Metab 2013;63(suppl 1):1–1960

PO2683
DETERMINATION OF THE PHENOLIC COMPOSITION OF GRAPE JUICE AND DIFFERENT TYPES OF
WINES FROM MONTEVIDEO AND INTERIOR OF
URUGUAY
M. Panzl1, F. Noguera1

1
Department of Nutrition, Catholic University of
Uruguay, Montevideo, Uruguay
Background and objectives:: Several epidemiological studies have examined the relationship between the consumption
of red wine and its derivatives rich in phenolic compounds and
cardiovascular disease. That is why we propose the following
problem: if the wine contains polyphenols, anthocyanins in
them and these compounds provide health benefits, Can grape
juice polyphenols present and provide similar benefits? The objectives: Determination of the phenolic composition in grape
juice and different types of wines (red, white and pink) from
the departments of Canelones, Montevideo and Paysandú in
the June-July period of 2011.
Methods: A descriptive cross with a type of non-probability
sample of convenience with a total of 67 samples from 20 local wineries. Each sample was analyzed in the total polyphenol
index (IPT) and total anthocyanins (At), using the methods of
ultraviolet absorbance at 280 nm and 520 nm (nanometers).
Results: The group of red wines showed the highest values
in the quantification of total phenolics and total anthocyanins
(76.79 and 655.33 mg / l), then follows the grape juice (32.36
and 175.88 mg / l). Thirdly rosés group (14.89 and 74.46 mg /
l), and the white wine group has the lowest values (11.90 and
3.28 mg / l).
Conclusions: The grape juice could be considered a food
source of antioxidants, which contributes to health, due to the
polyphenolic compounds content.
Key words: antioxidant, phenolic compounds, juice.

PO2684
INCREASING THE RESISTANT STARCH CONTENT
OF CHICKPEA FLOUR BY ENZYMATIC AND THERMAL TREATMENTS

A. Guadix1, R. Morales-Medina1, M. Munio1, P. Gonzalez-Tello1, F. Camacho1, E M. Guadix1
1
Department of Chemical Engineering, University
of Granada, Spain
Background and objectives: Resistant starch (RS) is the
starch which can not be absorbed in the small intestine of
healthy humans. RS has positive effects on diabetes, cardiovascular diseases, colonic health, obesity and osteoporosis.
Abstracts


Considering potential sources of RS, the scientific literature
has reported that the starch contained in legumes can be transformed into RS by means of enzymatic and thermal methods.
The purpose of this work was to compare the efficiency of both
methods intended to increase the RS content of chickpea flour.
Methods: In the case of enzymatic treatment, a suspension
of chickpea flour was pre-heated at 100 ºC for 15 min. Then,
the suspension was hydrolyzed with pullulanase (EC. 3.2.1.41)
for 24 h. In the case of thermal treatment, 4 heating-cooling
cycles were applied. Each cycle comprised heating at 100 ºC
for 30 min plus cooling at 4 ºC for 24 h. The RS content of
the lyophilized samples was determined using the AACC Official Method 32-40, which included a simulated digestion with
α-amylase at 37 ºC for 16 h.
Results: The initial RS value of the flour (0.88%) improved
significantly by the usage of both treatments. Regarding the enzymatic one, after the pre-heating step, the RS amount reached
4.47%. This value was further enhanced up to 7.47% at the end
of the hydrolysis. By the usage of the thermal treatment, the
RS content increased up to 8.74%, 9.85%, 10.28% and 10.67%,
after 1, 2, 3 and 4 cycles, respectively.
Conclusions: Thermal as much as enzymatic treatments
were effective in order to enhance the amount of RS which is

present in raw chickpea flour. Due to hydrolysis with pullulanase, RS content was multiplied by a factor of 8.5, while executing 4 heating-cooling cycles provided a factor of 12.

a simulated digestion process, which consisted in two consecutive hydrolysis with pepsin (1 hour, 37 ºC, pH 2) and pancreatin (2 hours, 37 ºC, pH 7.5). The ACE inhibitory activity
of hydrolysates was evaluated before and after the simulated
digestion by a spectrophotometric assay using 2-furanacryloylL-phenyl-alanylglycylglycine as substrate.
Results: The values of the IC50 (defined as the concentration of hydrolysate which reduces the ACE activity by 50%)
of digested hydrolysates were compared with the values of the
initial samples (248 microg/L). When the complete simulated
digestion was performed, an increase of 3% in the IC50 only
was measured in the case of the subtilisin hydrolysate. For the
trypsin hydrolysate, it was reached the highest worsening in
the IC50 (53%). Finally, when the mixture of both enzymes was
employed, an intermediate increase (16 %) in the IC50 was observed.
Conclusions: The hydrolysis of goat milk caseins with subtilisin, trypsin or a mixture of both enzymes yielded hydrolysates which maintained their ACE inhibitory activity to some
extent after being subjected to simulated digestion with pepsin
and pancreatin.

Key words: resistant starch, simulated digestion, legume,
chickpea.

PO2686
AZUKI(RED) BEANS PASTE: EFFECT OF COOKING
TIME ON RESISTANT STARCH CONTENTS AND
PARTICLE MORPHOLOGY
A. Kamei1, R. Atsumi1
1
Division of Domestic Science Education, Miyagi
University of Education, Sendai, Miyagi prefecture,
Japan


PO2685
INFLUENCE OF SIMULATED DIGESTION IN THE
ACE INHIBITORY ACTIVITY OF GOAT MILK PROTEIN HYDROLYSATES
A. Guadix1, F J.Espejo-Carpio1, M C. Almecija1, R. PerezGalvez1, P J. Garcia-Moreno1, E M. Guadix1
1
Department of Chemical Engineering, University of Granada, Spain

Background and objectives: Some peptides derived from
food proteins have shown angiotensin converting enzyme
(ACE) inhibitory activity, which can be useful in the formulation of functional foods intended for hypertensive individuals.
However, these peptides must remain active after the digestion
process in order to exert its inhibitory action. The objective of
this work was to evaluate the effect of simulated digestion on
the ACE inhibitory activity of protein hydrolysates from goat
milk.
Methods: Goat milk casein hydrolysates were produced by
three different enzymatic treatments, using subtilisin, trypsin
and the mixture of both. The hydrolysates were summited to
Abstracts

Key words: Bioactive peptides, ACE-inhibitory activity,
protein hydrolysates, goat milk, simulated digestion.

Background and objectives: Traditional Japanese sweets
are very popular and are often eaten by middle-aged and elderly
people. A lot of traditional Japanese sweets are made using an
Azuki bean paste, so called ‘An’ in Japanese. Azuki beans are a
good source of carbohydrate as well as protein, because they
are starchy pulse. Resistant starch (RS) escapes digestion until
reaching colon and acts like dietary fiber. Recently, many researchers suggest taking this type of dietary fiber for our health

benefits. The purpose of this study was to investigate the effect
of different cooking times on the amount of RS and the morphology of ‘An’ particles.
Methods: Azuki beans were boiled 50, 70, 90 minutes in
water. The ratio of Azuki beans to water was one to five. After
grinding, each mixture was strained through a sieve in order to
remove husk and put into cheesecloth. Then, a 6kg of stone was
placed on cheesecloth for 1 hour to discard excess water and
to make ‘An’. The amount of RS of each ‘An’ was measured and
each ‘An’ particles was observed by optical microscope.
Ann Nutr Metab 2013;63(suppl 1):1–1960

1563


Results: The amount of RS of ‘An’ in 50, 70, 90 minutes
cooking time were 6.4%, 5.0%, 4.4%, respectively. This result showed that increasing cooking time was decreasing the
amount of RS. Optical microscope observation showed that as
the cooking time become longer, the ratio of damaged or ruptured ‘An’ particles increased.
Conclusions: These results indicated that damaged ‘An’
particles had more digestible starch than intact ‘An’ particles.
One of the possible reasons could be that intact ‘An’ particles
resist digestive enzymes. Another reason might be that a starch
in ‘An’ particles affects its structure during cooking.
Key words: Azuki bean paste ‘An’, resistant starch, ‘An’ particles

tion of DPPH radical and β-Carotene bleaching. Extracts of C.
bonariensis and P. granatum showed inhibitory effects on the
gram positive and negative bacteria at all added doses.
Conclusions: Extracts of seven Yemeni plants showed antioxidative and antibacterial activities. Extracts of C. bonariensis
and P. granatum possessed the best activities may attribute to

their high content of polyphenols.
Key words: Total phenolic, Antioxidant , Antibacterial,
Conyza bonariensis, Punica granatum

PO2687
EXTRACTS OF SENEN YEMENI PLANTS WITH ANTIOXIDATIVE AND ANTIBACTERIAL ACTIVITIES
R A S. Thabit1, X R. Cheng1, X. Tang1, J. Sun1, Y H. Shi1, G
W. Le1
1
State Key Laboratory of Food Science and Technology, School of Food Science and Technology,
Jiangnan University, Wuxi, China

PO2688
ALGINATE ENRICHED BREAD AS AN OBESITY
TREATMENT
D. Houghton1, M. Wilcox1, I. Brownlee2, C. Seal3, J. Pearson1
1
ICaMB, Newcastle University, Newcastle Upon
Tyne, NE2 4HH, UK
2
Food and Human Nutrition Department, Newcastle University office, 180 Ang Mo Kio, Singapore
3
Human Nutrition Research Centre, School of Agriculture, Food and Rural Development, Kings Road,
Newcastle upon Tyne, UK

Background and objective: The increasing interest in natural dietary components has focused attention on plants used as
food or spices which are a rich source of bio-nutrients or bioactive phytochemicals. The most important of these bioactive
compounds of plants are alkaloids, tannins, and phenolic compounds. Herbs and spices have been employed since ancient
times as flavouring and storing agents for food, attributing to
their antimicrobial and antioxidant components. This study

aims to find out the phenolic contents of the most important
plants grown in Yemen and their antioxidative and antibacterial activities.
Methods: The total contents of phenolic (TPC), flavonoid
(TFC), tannins (TTC), and anthocyanin (TAC) in the extracts
of seven Yemeni plants: Punica granatum, Zizyphus vulgaris,
Ruta graveolens, Lavandula officinalis, Thymus vulgaris, Conyza bonariensis, and salavia officinalis were determined by Folin-Ciocalteu method, flavonoid-aluminium complex method,
Folin and Ciocalteu method, and pH-differential method. The
antioxidative activities of the extracts were determined by
DPPH and β-carotene bleaching. The effects of the extraction
on the growth inhibition of some indicators of foodborne illness bacteria were investigated.
Results: There is a great variability in the TPC, TFA, TTC,
TAC, inhibition of DPPH radical and β-Carotene bleaching of
the extraction obtained from the seven plants. C. bonariensis
and P. granatum presented the best antioxidative activities, had
the highest TPC, TFC, TTC, and TAC and strongest inhibi-

Background and objectives: Obesity is a fast growing medical issues worldwide is fast becoming one of the leading causes of mortality. Data from our laboratory have demonstrated
that alginate possess the ability to inhibit pancreatic lipase invitro. The purpose of the present study was to determine if alginate enriched bread inhibits fat digestion in-vitro and in-vivo.
Methods: Control bread and alginate bread developed by
Greggs Plc was used. Trioctanoate and tributyrate were digested in an open model gut system developed within our laboratory which replicates digestion in the mouth, stomach and
small intestines. The substrate was added to the model gut alone or with either 5.2 g alginate bread or 5.2 g control bread
(n=6). 1ml samples were taken every 15 minutes from 0-180
minutes and analysed for free glycerol. Preliminary data from
a double blind study using ileostomy patients (n=13) fed either
an alginate bread (105 g) or control bread (105 g) (visits were
one month apart), which included 20 g of butter post fasting.
Blood samples were taken at baseline and then every 30 minutes. A t-test was used to compare control bread and alginate
bread with normal substrate digestion, and to compare total
triglycerides in plasma between alginate and control bread.
Results: At 180 minutes there was a significant reduction

in fat digestion of 47.7% (1.2) and 32.5% (2.4) for trioctanoate
(p>.05) and tributyrate (p>.05) respectively. Data from ileostomy patients indicated a reduction in the total triglycerides
in the plasma ranging from 9-34% compared with the control
bread, with a significant difference at 210 minutes (p>.05).

1564

Ann Nutr Metab 2013;63(suppl 1):1–1960

Abstracts


Conclusions: This study provides evidence that normal
foods supplemented with alginate could be used to treat obesity/overweight. Further analysis is required of a larger group
of ileostomy patients and to assess the ileum effluent fluid to
demonstrate undigested lipids.

high-fat feeding, together with the weight loss induced in rats,
allow this polyphenol to be proposed as a therapeutic agent in
the chronobiological treatment of obesity.
Key words: High-fat feeding, Clock gene,: Resveratrol, Rats

Key words: obesity, digestion, alginate.

PO2689
EFFECTS OF RESVERATROL ON CHANGES INDUCED BY HIGH-FAT FEEDING ON CLOCK GENES IN
RATS
J. Miranda1,2, M P. Portillo1,2, J A. Madrid3, N. Arias1,2, M T.
Macarulla1,2, M. Garaulet3
1

Nutrition and Obesity Group, Dpt. Nutrition and
Food Sciences, Faculty of Pharmacy, University of
the Basque Country (UPV/EHU), Vitoria, Álava, Spain
2
CIBERobn Physiopathology of Obesity and Nutrition, Institute of Health Carlos III (ISCIII), Madrid,
Spain
3
Dpt. Physiology, Faculty of Biology, University of
Murcia, Murcia, Spain
Background and objectives: In mammals, the main component of the circadian system is the suprachiasmatic nucleus
of the hypothalamus, but circadian clocks are also present in
most peripheral tissues such as adipose tissue or liver. The purpose of the study was to analyze the potential effects of resveratrol on changes induced by high-fat feeding in the expression
of different clock genes and clock-controlled genes (CCG) in
white adipose tissue and liver from rats.
Methods: 24 rats were divided into three groups: the control group fed a standard diet and two groups fed a high-fat
diet supplemented with resveratrol (RSV) or not (HF). The polyphenol was daily added to the diet in amounts that assured a
dose of 30 mg/kg body weight/d. The expression of clock genes (Bmal1, Clock, Npas2, Cry1 and Per2) and CCGs (PPARγ,
Rev-Erbα, Rorα and Sirt1) was analyzed by real time RT-PCR
in epididymal adipose tissue and liver.
Results: Final body weight, adipose tissue weights and liver
weight were significantly higher in animals from HF groups
than those from control group. Resveratrol reduced body
weight and the sum of adipose tissues but these parameters did
not reach the values observed in control rats. By contrast, resveratrol did not modify liver weight. When compared with the
controls, the RSV group showed similar patterns of response to
HF group, except for Rev-Erbα, which was down-regulated in
both analyzed tissues.
Conclusions: The effects of resveratrol on changes induced
on the circadian clock machinery in adipose tissue an liver by
Abstracts


PO2690
MALAYSIAN SELECTED LOCAL BASED DISHES:
CONTENTS OF MACRONUTRIENT, ISOFLAVONE,
TOTAL PHENOL AND ANTIOXIDANT ACTIVITY.
H. Haron1, L. Siew Fen1, Y. Zu Wei1,
1
Nutrition Science Programme, School ofHealth
Care, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia
Background and objectives:Phenolic content in soybean
have been identified to prevent chronic diseases through antioxidative activity. This study was carried out to determine the
macronutrient, total phenol, isoflavone contents and anti-oxidative activity in selected soy based dishes.
Methods: Eight types of local soy based dishes were bought
from restaurants and food stalls located in Kuala Lumpur. Each
sample from two different locations were analysed in duplicates. Proximate contents were analysed by using AOAC methods
(1997) while carbohydrate content was calculated by difference.
Isoflavone content was analysed using High Performance Liquid Chromatography method by Hutabarat et al. (2000). Total
phenol content (TPC) was determined using Folin-Ciocalteu
method while antioxidant activities were assessed using DPPH
radical scavenging assay and B-carotene bleaching assay.
Results: The soy based dishes contained 8.18-161.34 mg
total isoflavone/ 100g food sample, 34.91-87.64% moisture,
3.93-13.83% protein, 4.35-32.90% fat, 0.53-41.74% total carbohydrate and 1.20-1.92% ash. Average TPC in all the dishes was
in the range of 19.14-62.82 mg of GAE/g wet weight. Antioxidant activities based on DPPH radical scavenging assay and Bcarotene bleaching assay were significantly (p <0.01) correlated
with TPC. Total isoflavone content was significantly (p < 0.01)
correlated with TPC (r=0.711), DPPH radical scavenging assay (r=0.720) and B-carotene bleaching assay (r=0.720). Dishes
containing tempeh has significantly (p < 0.05) higher isoflavone content compared to other dishes containing other type of
soy products.
Conclusions: Soy product like egg tofu contained highest
moisture but lowest protein and isoflavone contents. Method of

cooking such as frying increased the fat content in the dishes.
Dishes containing fermented food such as tempeh have high
amount of isoflavone and significantly higher antioxidant activity compared to other type of soy products.
Key words: Soy based dishes, Isoflavone, Total phenol content, Macronutrient, Antioxidant activity.
Ann Nutr Metab 2013;63(suppl 1):1–1960

1565


PO2691
COMPARATIVE ANALYSIS OF VOLATILES FROM
PLANT CZ BY HDE-GC/MS
K. Chang1, G. Kim1
1
Plant Resources Research Institute, Duksung
Women’s University, Korea
Background and objectives: The purpose of this report was
to evaluate and compare the chemical variability of the volatile
compositional differences of CZEOs in Korea. Volatile composition differences of CZEOs prepared via different producing
methods (wild-growing and cultivate-growing) were also determined.
Methods: The dried CZ were crushed for 10 sec by a blender
(NJ-8060SM, NUC Electronics, Seoul, Korea) and extracted by
hydro-distillation extraction (HDE) method for 3 hr using a
Clevenger-type apparatus (Hanil Lab Tech Ltd, Incheon, Korea). and analyzed by gas chromatography-mass spectrometry
(GC/MS).
Results: Total 57 and 55 volatiles constituents were confirmed in CZEOs from wild-growing and cultivate-growing. Alcohols were predominant in the volatiles of CZ flowers (%): the
wild, 37.57; and the cultivated, 30.34.
Conclusions: There was some differences between CZEOs
from wild-growing and cultivate-growing; the yields of CZEOs
were 0.35 and 0.12%(v/w), respectively. Borneol was the most

abundant volatiles in the wild-growing and Ą-curcumene was
the major component in the cultivated.
Key words: volatile composition, producing methods, hydro-distillation extraction borneol, Ą-curcumene

PO2692
EFFECTS OF SERICULTURAL PRODUCTS ON
BLOOD GLUCOSE AND LIPID STATUS IN STREPTOZOTOCIN-INDUCED DIABETIC MICE
S H. Cho1, S W. Choi1
1
Department of Food Science and Nutrition, Catholic University of Daegu, Gyeongbuk, Korea
Background and objectives: The effects of four types of
sericultural products on the blood glucose and lipid status as
well as peroxidative state and muscle glycogen level were investigated under diabetic condition. The four products were
silkworms (SW), mulberry leaves (ML), mulberry fruit (MF),
and native mulberry fruit (kujippong; NMF).
Methods: Dried powders of the four products were added
to diets (AIN-93G) for the corresponding four groups of
streptozotocin-induced diabetic mice and fed for four weeks.
Non-diabetic (Normal) and diabetic mice fed no test product
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Ann Nutr Metab 2013;63(suppl 1):1–1960

(DM-Control) were another experimental groups to compare
the effects of test diets.
Results: Compared to the DM-Control (mean; 289.4±23.1
mg/100mL), the NMF group (mean; 255.3±19.9 mg/100mL)
increased blood glucose at reduced rate most effectively
among the test groups. Serum cholesterol and triglyceride levels were lowest in the ML group (114.2±17.5 and 136.9±14.4
mg/100mL) but those of SW, MF, and NMF groups were tended to be lower than the DM-Control group (159.6±14.7 and

169.0mg±15.1 mg/100mL). Liver cholesterol and triglyceride
contents were lower in the MF (12.8±1.2 and 12.9±1.9 mg/g)
and the NMF groups (12.9±1.0 and 13.2±1.5 mg/g) than the
DM-Control group (16.5±1.3 mg/g, and 21.1±2.3 mg/g). Liver level of thiobarbituric acid reacitve substances (TBARS)
was lower in the NMF (20.9±1.9 nmol MDA/g) than the other
groups while muscle glycogen level was higher only in the MF
group than the DM-Control group.
Conclusions: A proper combination of mulberry fruit, kujippong, and mulberry leaves as the ingredients of functional
foods helps blood glucose and lipid status maintain proper levels and improves body peroxidation under diabetic state.
Key words: mulbery, blood glucose, lipid, diabetic mice

PO2693
THAI FRUIT SOURCES FOR LUTEIN AND
ZEAXANTHIN : PROTECTIVE EFFECTS AGAINST
AGE-RELATED MACULAR DEGENERATION (AMD)
N. Vatanasuchart1, P. Khaiprapai2, U. Phukasmas1
1
Department of Nutrition and Health, Institute of
Food Researchand Product Development, Kasetsart
University, Bangkok, Thailand
2
Food Quality Assurance Services Center, Institute
of Food Researchand Product Development, Kasetsart University, Bangkok, Thailand
Background and objectives: Lutein and zeaxanthin play an
important role in protecting against age-related macular degeneration (AMD), rising an evidence in the elderly. This study
was aimed at determining lutein and zeaxanthin in 170 Thai
fruits.
Methods: Fruit sample was extracted with 0.1% BHT in
MeOH and analyzed by HPLC.
Results: It showed that lutein was predominately found in

Thai fruits about 122 samples or 71.76% of total samples, while zeaxanthin was rarely found about 65 samples or 38.24 %.
When fruits were classified according to sugar content, Thai
melon (Tangthai) contained the highest lutein of 66.17 ± 26.4
µg/100g in group of sugar < 10%. Among all samples, Tangthai also provided the highest lutein, following with roseapple
(Phed), watermelon (Torpedoe and Kinaree), guava (Kimju),
Abstracts


mango (Pramkaymea) and pamelo (Kaonumpueng), respectively. In group of sugar between 10-20%, banana (Namwa) contained the highest lutein of 26.04 ± 6.76 µg/100g following with
other banana varieties and sapote (Lamod). In group of sugar >
20%, all jackfruits contained lutein and Sritong variety had the
highest lutein of 22.91 ± 8.25 µg/100g. Whereas ripe mangoes
and sweet tamarinds did not contain lutein. For determination
of zeaxanthin, mango (Munkunsri), in group of sugar < 10%
had the highest of 24.93 ± 49.86 µg/100 g, following with papaya (Kekdum), mango (Keowsavoy), pamelo (Kaonumpueng),
mango (Pramkaymea) and pamelo (Tongdee), respectively. In
group of sugar between 10-20%, Maprang showed the highest
zeaxanthin of 27.29 ±26.43 µg/100 g, followed with Mayongchid, mango (keowyai), orange, pineapple and starfruit, respectively. Source of both lutein and zeaxanthin were Tangthai,
pamelo, roseapple, raw mangoes, Maprang, Mafueng, orange,
sapote, purple passionfruit and pineapple.
Conclusions: Thai fruits are important source of lutein and
zeaxanthin which should be regularly consumed to promote
eye health, particularly for the fruits with low sugar.
Key words: Thai fruits, lutein,zeaxanthin, eyehealth.

PO2694
ADLAY CONSUMPTION AMELIORATED THE PROGRESSION OF ALCOHOLIC LIVER DISEASES
H Y. Yang1, W J. Yeh2, H Y. Weng2, C Y. Chein2, J R. Chen2
1
I-Shou University, Kaohsiung, Taiwan

2
Taipei Medical University, Taipei, Taiwan
Background and objectives: Fatty liver has become a
health problem in Taiwan and dietary modification may show
beneficial effects. Adlay has been reported to exhibit hypolipidemic, antiobesity and antiinflammation effects. Therefore, the
aim of the study is to investigate effects of adlay on alcoholic
liver disease.
Methods: We fed rats with ethanol liquid diet to induce alcoholic liver disease (group E) and added 22.1 g/L (group A)
and 44.2 g/L adlay (group B) powder in diet of rats. After a
12-wk experimental period, rats were sacrificed for blood and
liver sample collections.
Results: We found elevation of hepatic index AST and ALT
in the ethanol group since the 4th week and adlay consumption
attenuated the elevation of hepatic index. At the end of the study, the ethanol group showed hepatic steatosis and higher levels of liver malondialdehyde, tumor necrosis factor-and early
fibrosis markers hydroxyproline than the control group and
the two adlay groups. We also found that group A had lower
cytochrome P450 2E1 protein expression than group E. In the
histological analysis, the fatty change scores of both central and
portal vein zone of group A were also lower than group E.
Abstracts

Conclusions: These results suggested that dietary adlay
consumption may be beneficial in the prevention of alcoholinduced hepatic fat accumulation and inflammation.
Key words: alcohol, fatty liver, adlay, inflammation

PO2695
BENEFICIAL EFFECTS OF SOY PROTEIN AND
β-CONGLYCININ ON NONALCOHOLIC LIVER DISEASES VIA AMELIORATING HEPATIC STEATOSIS
AND INSULIN RESISTANCE
W J. Yeh1, C Y. Chien1, H Y. Weng1, J R. Chen1, H Y. Yang2

1
Taipei Medical University, Taipei, Taiwan
2
I-Shou University, Kaohsiung, Taiwan
Background and objectives: Nonalcoholic fatty liver disease (NAFLD) is the most common liver disorder in the Western
world and is related to obesity, insulin resistance, inflammation,
and oxidative stress. Nonalcoholic steatohepatitis (NASH) is
the most extreme form of NAFLD, and is characterized by steatosis with necroinflammation. This study aims to explore the
beneficial effects of soy protein and soy-conglycinin on hepatic
steatosis and insulin resistance in NASH.
Methods: Male Wistar rats were induced to have NASH
with high-fat-liquid diet for 16 weeks and divided into NASH
group (N), soy protein group (S), and β-conglycinin group (B).
Experimental diets were based on high-fat-liquid diet with
50% soy protein or equivalent β-conglycinin as substitution of
casein for 8 weeks. Plasma and liver samples were collected for
analysis of lipid profiles, cytokines and insulin sensitivity, as
well as cytochrome P-450 2E1 (CYP2E1) protein expression
and pathological analysis in the liver. Feces were assembled for
lipid content analysis.
Results: Rats treated with high-fat diet for 16 weeks showed
abnormal plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity, and they also exhibited
increasing levels of TG, interleukine(IL)-1and MDA concomitant with an upregulation of CYP2E1 in the liver. After 8-wk
soy protein or β-conglycinin consumption, hepatic TG, tumor
necrosis factor (TNF)-and MDA concentrations were lower
than those in the NASH group, whereas the systemic insulin
sensitivity was also improved by β-conglycinin. We also found
improvement of hepatic steatosis in pathological analysis.
Conclusions: Soy protein and soy β-conglycinin may improve the liver function in rats with NASH by lowering lipid
levels in the liver and ameliorating inflammation and insulin

resistance.
Key words: soy protein,conglycinin, fatty liver, nonalcoholic steatohepatitis

Ann Nutr Metab 2013;63(suppl 1):1–1960

1567


PO2696
GREEN TEA EXTRACT SUPPRESSES DISUSE MUSCLE ATROPHY IN MICE
H. Tachibana1, M. Murata1, R. Kosaka1, K. Kurihara1, A.
Nesumi2, M. Maeda-Yamamoto3, K. Yamada1
1
Kyushu University, Faculty of Agriculture, Japan
2
National Institute of Vegetables and Tea Science,
NARO, Japan
3
National Food Research Institute, NARO, Japan
Background and objectives:A variety of stresses, including
starvation, muscle disuse, systemic illness, and aging cause
skeletal muscle atrophy. Loss of muscle protein primarily is a
result of enhanced protein breakdown, which is initiated by the
activation of the ubiquitin-proteasome pathway. Two musclespecific E3 ubiquitin ligases, muscle RING finger 1 (MuRF1)
and muscle atrophy F-Box (Atrogin-1) are recognized as markers of muscle atrophy since they are expressed at relatively low
levels in resting muscle and transcriptionally increased under
a variety of atrophy-inducing conditions. In this study, we examined the suppressive effects of green tea extracts on disuse
muscle atrophy and its causative genes in mice.
Methods:Tail suspension was maintained for 10 days using
the male C57BL6 mouse, and the oral administration of green

tea (Sunrouge or Yabukita) extract continued at 48 h intervals
during the tail suspension. Gastrocnemius, soleus, plantaris,
and quadriceps femoris were extracted, and weight was measured. In each muscles, the expression level of MuRF1 and Atrogin-1 gene was measured.
Results: The tail suspension group had a significantly decreased ratio of gastrocnemius, soleus, plantaris, and quadriceps femoris weight as compared with the ground group. Oral
administration of Sunrouge or Yabukita extract suppressed the
decrease of muscle weight ratio in the tail suspension group.
The tail suspension induced mRNA expression of MuRF1 and
Atrogin-1The green tea extracts significantly suppressed the induction of MuRF1 and Atrogin-1 in the tail suspension group.
Conclusions: The green tea has a potential to attenuate skeletal muscle atrophy. The inhibitory effect of the green tea may
be due to the suppression of the ubiquitin ligases MuRF1 and
Atrogin-1 induced by muscle atrophy.
Key words:Green tea, Muscle atrophy, MuRF1, Atrogin-1,
Stress

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Ann Nutr Metab 2013;63(suppl 1):1–1960

PO2697
PHENOLIC CHARACTERIZATION AND ANTIOXIDANT ACTIVITY OF STRAWBERRY (FRAGARIA
ANANASSA) PUREE
M C. Garcia-Parrilla1, M A. Alvarez-Fernandez1, A B. Cerezo1, R. Hornedo-Ortega2, A M. Troncoso3
1
University of Sevilla, Sevilla, Spain
Background and objectives:Strawberry is harvested in a
very short period of time involving a fruit surplus frequently.
Furthermore, it is a very perishable fruit that diminish the possibilities of marketing. The production of strawberries puree
can be used to prevent the fruit losses increasing the conservation period.This work aims to establish the impact of industrial
processing on the phenolic composition and antioxidant activity of strawberry purees analyzing their evolution along the
production process.

Methods:The samples used for the present study were
strawberry purees of two consecutives years (2011 and 2012).
We set up and validated an analytical Liquid Chromatography
with Diode Array Detector (LC-DAD) method confirming the
identification by Mass Spectrometry. In vitro antioxidant activity was determined by ORAC and DPPH methods.
Results: This work was achieved reporting malonyl caffeoylquinic acid for the first time in strawberry. (+) Catechin
was the most abundant phenolic compound. The most abundant flavonol was kaempferol glucoside derivate.These results
show that industrial processes does not break the seeds to release the phenolic compounds as could be expected and purees
with or without seedless present a similar composition.
Conclusions: Strawberries purees are a good source of phenolic compounds with antioxidant potential, being of interest
from a nutritional and commercial perspective as ingredients
or raw material for further derived food.The antioxidant capacity decreased during the pasteurization step just slightly and
no important changes in terms of antioxidant activity are detected throughout the whole process.
Acknowledgements: The authors are grateful for the financial assistance provided by the Spanish Government (Project
MICINN AGL2010-22152-01). We are also grateful to HUDISA Desarrollo Industrial S.A., in Lepe, Spain, for providing the
strawberry samples.
Key words: phenolic compounds, strawberry, HPLC, antioxidant activity, ORAC, DPPH.

Abstracts


PO2698
TOCOPHEROL CONTENT AND TOTAL ANTIOXIDANT CAPACITY IN EXTRA VIRGIN ARGAN OIL
C. Cabrera-Vique1, A. Rueda1, C. Samaniego-Sánchez1,
R. Giménez1, M. Olalla1, I. Seiquer2
1
Departamento de Nutrición y Bromatología, University of Granada, Granada, Spain
2
Instituto de Nutrición Animal, Consejo Superior de
Investigaciones Científicas (CSIC), Granada, Spain

Background and objectives: Virgin argan oil has been
traditionally consumed in the Magreb countries, but recently
its consumption has been extended into European, Japan and
USA oil-markets. This oil is rich in oleic and linoleic acids (45
and 34%, respectively) and tocopherols. Polyphenols, sterols,
carotenoids, squalene, and other minor compounds also contribute to its antioxidant capacity. However, available bibliography on composition data is scarce and contradictory due to the
lack of standarization of the oil extraction method. In this study, we selected extra virgin argan oil to determine tocopherols
and total antioxidant capacity (AC) in comparison with other
edible vegetable oils widely consumed.
Methods: The presence of alfa, beta, gamma and deltatocopherols were quantified by HPLC following the UNE-EN
ISO 9936-2006 method. AC was determined by ABTS, DPPH
and FRAP methods previous sample treatment with a mixture
methanol/water.
Results: Total tocopherols in virgin argan oil ranged between 342.3–510.6 mg/kg, being gamma-tocopherol the major
fraction (78.3%); alfa, beta and delta-tocopherols represents
11.2, 2.1 and 8.5%, respectively. The mean content in extra
virgin olive oil (Picual variety) was 244.1 mg/kg and alfa-tocopherol was the major fraction. In virgin argan oil, the AC varied between 0.099-1.052, 0.056-0.257 and 0.179-0.237 mmol
Trolox/kg for ABTS, FRAP and DPPH methods, respectively.
Statistically significant higher AC values were detected in virgin olive oil but not in other oils such as soy or sunflower oils.
Nosignificant correlation was observed between tocopherol
content and AC.
Conclusions: The elevated presence of gamma-tocopherol
(a potent antioxidant and anti-inflammatory agent) and its major total AC in relation to other oils corroborates the extra virgin argan oil nutritional value and its potential human healthy
properties. Present findings may provide additional market
opportunities for the food industry to develop new products.
Key words: argan oil, tocopherols, antioxidant capacity,
edible oils

Abstracts


PO2699
BIOACCESIBILITY OF POLYPHENOLS AND ANTIOXIDANT ACTIVITY IN VIRGIN ARGAN OIL. COMPARISON WITH VIRGIN OLIVE OIL
C. Cabrera-Vique1, A. Rueda1, R. Gimenez1, M. Olalla1,
I. Seiquer2
1
Departamento de Nutrición y Bromatología, University of Granada, Granada, Spain
2
Instituto de Nutrición Animal, Consejo Superior de
Investigaciones Científicas (CSIC), Granada, Spain
Background and objectives: Phenolic compounds are of
essential importance in the nutritional properties of vegetable
oils, mainly related with their potent antioxidant activity. Nevertheless, to achieve any in vivo effect these bioactive compounds must be bioaccesible, which depend on their release
from the food matrix during the digestion process. The aim of
this study was to evaluate the bioaccesibility of total phenols
and antioxidant properties of extra virgin argan oils (AO) as
compared with extra virgin olive oil (OO), using an in vitro
gastrointestinal digestion method to approach physiological
conditions. Results were compared with those obtained in oil
extracts before digestion process.
Methods: Chemical extraction of antioxidants from oils
was performed by addition of methanol/acetone/water. Moreover, oil samples were in vitro digested in two sequential
steps to simulate gastric and duodenal conditions, followed by
centrifugation. Chemical extracts and supernatants of digested
samples (which represent bioaccesible fraction) were used to
determine phenolic compounds (Folin-Ciocalteau method)
and antioxidant activity (ABTS and FRAP assays).
Results: Phenol content and antioxidant properties were
always higher in bioaccesible oil fractions than in chemical extracts. Phenol content in chemical extracts was lower in AO
(ranging 193-624 mg/kg) than in OO (1018 mg/kg), but only
minor differences were observed in bioaccesible fractions (AO

2440-3008 mg/kg, OO 3618 mg/kg). A similar tendency was
observed for the antioxidant activity, and high correlations
were shown between phenol content and ABTS and FRAP assays (p<0.001).
Conclusions: Differences of polyphenol content and antioxidant activity found between AO and OO in chemical extracts
were attenuated in the bioaccesible fraction obtained after the
in vitro digestion process, as comparatively higher release of
antioxidants due to digestive enzymes activity seem to occur
in AO.
Key words: virgin argan oil, polyphenols, antioxidant activity, bioaccesibility.

Ann Nutr Metab 2013;63(suppl 1):1–1960

1569


PO2702
THE NATURAL IMINUSUGAR D-FAGOMINE REDUCES WEIGHT GAIN AND FAECAL ENTEROBACTERIAL POPULATION OF RATS FED AN ENERGYDENSE DIET
S. Ramos-Romero1, E. Molinar-Toribio1, J. Perez-Jimenez1, M. Casado2, B. Pina2, J L. Torres1
1
Department of Biological Chemistry and Molecular Modelling, Institute of Advanced Chemistry of
Catalonia (IQAC-CSIC), Barcelona, Spain
2
Department of Environmental Chemistry, Institute
of Environmental Assessment and Water Research
(IDAEA-CSIC), Barcelona, Spain
Background and objectives: Increasing evidence reveals
that diet induced changes in the composition of gut microbiota
are related to fat accumulation, insulin resistance and obesity.
Particularly, high consumption of fat and sucrose increases the
proportion of intestinal Enterobacteriales. As D fagomine, an

iminosugar analogue of D glucose, agglutinates Enterobacteriaceae and inhibits their adhesion to intestinal mucosa in vitro, we hypothesized that it may have a functional effect on the
gut microbiota of animals fed an energy-dense diet.
Methods: Rats were fed a standard diet, a high-fat, high sucrose diet and a high-fat, high sucrose diet supplemented with
D fagomine (0.065 %) for 5 weeks. Body weight and feed intake
were measured at the onset and at the end of the study. Total
bacteria, Enterobacteriales and Escherichia coli were quantified by quantitative real-time PCR on faecal DNA.
Results: During the nutritional intervention energy intake
was independent of the diet. After 5 weeks the mean weight
gain of the group fed the high fat, high sucrose diet was~13
% higher than that of the standard group whereas the gain in
the group supplemented with D fagomine was ~7 % higher
(P<0.05). All the animals presented similar amounts of total
bacteria in faeces. The relative abundance of Enterobacteriales and E. coli excreted was significantly higher (P<0.05) in the
high-fat, high-sucrose group. The animals supplemented with
D fagomine and the standard group presented similar amounts
of faecal Enterobacteriales and E. coli.
Conclusions: The iminosugar D fagomine reduces the
weight gain and the raise in the proportion of faecal enterobacteria associated with an energy-dense diet. The two effects
may be related.

PO2703
INHIBITORY EFFECTS OF PHENOLIC EXTRACTS
DERIVED FROM JAPANESE APRICOT FRUIT (PRUNUS MUME) ON RAT SMALL INTESTINAL DISACCHARIDASE ACTIVITIES
K. Kishida1, Y. Heya1, Y. Yamazaki1, A. Horinishi1, Y. Ozaki1
1
Department of Science and Technology On Food
Safety, Kinki University, Japan
Background and objectives: Japanese apricot fruit (Prunus
mume Sieb. et Zucc) has been traditionally used as a folk remedy in Japan.We have already established a preparation method
for extracting phenolics from ‘umezu’, exuding juice of salted

Japanese apricot fruits.Umezu phenolics (UP) contain about
12% (w/w) polyphenol as gallic acid equivalents and mainly
consist of hydroxycinnamic acids (caffeic acid, ferulic acid, pcoumaric acid) and their derivatives.Chlorogenic acid is one of
the most abundant phenolics in UP.The aim of this study was to
investigate whether UP inhibits small intestinal disaccharidase
activities and suppresses the postprandial elevation of blood
glucose levels in rats.
Methods: The disaccharidase (maltase, sucrase, isomaltase,
glucoamylase, lactase, trehalase) activities were assayed by the
Dahlqvist method using rat intestinal mucosa homogenates.A
single oral administration of 2 g/kg BW sugars (glucose, maltose, sucrose, starch) was carried out with or without UP and
blood glucose levels were measured up to 120 min.
Results: UP inhibited all disaccharidase activities examined
in this study although the effects varied among disaccharidases.Inhibitory effect of chlorogenic acid or non-esterified hydroxycinnamic acids at the concentration equivalent to those
in UP was relatively low.This may indicate the presence of other
unknown derivatives responsible for the inhibition. The blood
glucose levels were significantly altered when sucrose was administered to rats.The suppressive effects after other sugar load
tests were marginal.
Conclusions: These results suggest that UP could be useful
as a food ingredient to reduce risks for diabetes and obesity.
Key words: Japanese apricot, phenolics, disaccharidase,
blood glucose.

Key words: D-fagomine, iminosugars, obesity, microbiota,
Escherichia coli

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Ann Nutr Metab 2013;63(suppl 1):1–1960


Abstracts


PO2704
LACTOBACILLUS PARACASEI CBA L74 AND FERMENTED OAT INHIBIT ENTRANCE OF UNDIGESTED GLIADIN PEPTIDES IN CACO-2 CELLS
M. Sarno1, M. Nanayakkara1, M. Cuomo1, R. Nigro2, S.
Auricchio1, M V. Barone1
1
Department of Pediatrics and ELFID, University of
Naples Federico II, Italy
2
Department of Chemical Engineering, University
of Naples Federico II, Naples, Italy
Background and objectives: Recent reports describe a role
of probiotics as therapeutic approach for Celiac Disease (CD).
Undigested A-gliadin peptides P31-43 and P57-68 are central
to CD pathogenesis, inducing an innate and an adaptative immune response respectively. They enter enterocytes and localize to vesicular compartment to induce their toxic/immunogenics effects. Our aim is to test the effect of Heinz property
strain Lactobacillus Paracasei (LP) CBA L74 (International
Depository Accession Number LMG P-24778, its supernatant
and LP CBA L74 fermented oat on P31-43 and P57-68 entrance
in Caco-2 cells to verify itsprotective effect.
Methods: We treated Caco-2 cells with LP CBA L74, its colonies supernatant or with fermented oat for 30 minutes, and
then fluorochrome labeled P31-43 or P57-68 were added to
cells cultures. We studied entrance of labeled peptides by fluorescence assay.
Results: LP CBA L74 inhibit both P31-43 (FI: 75%) and
P57-68 (FI: 50%) entrance respect to control. LP CBA L74 supernatant is also able to induce decrease of both gliadin peptides entrance in Caco-2 cells (FI: 70% and 50% respectively),
indicating that this biological effect is due to some product included in LP CBA L74 supernatant. LP CBA L74 fermented oat
is also able to reduce P31-43 entrance in Caco-2 cells respect to
not fermented oat (FI: 50%).
Conclusions: LP CBA L74, its supernatant and fermented

oat, reduce P31-43 and P57-68 entrance in Caco-2 cells. This
is the first study that attempts to explain the molecular mechanism of probioticseffects in the prevention of undigested gliadin peptides toxic effects.

PO2705
IRON AND CALCIUM BIOACCESSIBILITY FROM
CEREAL DERIVATIVES
A. Cilla1, Y. Caldera1, R. Barberá1, M J. Lagarda1
1
Nutrition and Food Science Area, Faculty of Pharmacy, University of Valencia, Valencia, Spain
Background and objectives: Cereal flours and derivatives
are ideal foodstuffs for mineral fortification with iron (Fe) and
calcium (Ca). Iron deficiency is the most prevalent single nutritional deficiency in the world and is related to delayed cognitive development and intellectual impairment in children.
Calcium intake during childhood is associated with increased
peak bone mass during adolescence, bone fragility and hip
fracture prevention and a reduction in the prevalence of osteoporosis.The aim of this study was to determine the bioaccessibility (amount of mineral available for absorption and utilization) of Fe and Ca in six fortified biscuits and bakery products.
Methods: A simulated gastrointestinal digestion with two
stages (gastric with pepsin pH= 2 and intestinal with pancreatin and bile salts pH= 6.5) was applied to obtain the soluble
mineral (bioaccessible fraction) by centrifugation. Total and
bioaccessible iron and calcium were determined by atomic absorption spectrometry after dry mineralization (450ºC).
Results:Bioaccessible iron: 0.5-2 mg Fe/100g sample. The
highest Fe content was found in the product with one of the
lowest Ca/Fe ratios and one of the highest total Fe contents.
Negative influence: Ca content (Ca/Fe ratio) and fiber. Positive
influence: Fe content. Iron bioaccessibility was > 20% (23-32%).
Bioaccessible calcium: 49-79 mg Ca/100g sample. The highest
Ca content appeared in the product with less fiber content and
more total Ca. There was a trend of less soluble Ca when fiber
increases and in the presence of polyphenols used in the ingredients found in the formulation. Calcium bioaccessibility was
> 40% (40-70%).
Conclusions: Bioaccessibility results indicate that cereal

derivatives fortified with Fe and Ca are suitable complementary sources for meeting the nutritional requirements for these
minerals.
Key words: iron, calcium, bioaccessibility, cereal derivatives

Key words: probiotic, celiac disease, oat, LP CBA L74

Abstracts

Ann Nutr Metab 2013;63(suppl 1):1–1960

1571


PO2706
ANTIOXIDANT AND SCAVENGING CAPACITY OF
SERBIAN OAK TREE
B. Popovic1, D. Stajner1, R. Zdero1, S. Orlovic2, Z. Galic2,
M. Kebert
1
Faculty of Agriculture, Serbia
2
Institute of Lowland Forestry and Environment,
Serbia
Background and objectives: Quercus trees, commonly
known as oaks, belong to the family Fagaceae. They comprise
450 species worldwid. European oak corresponds well with these requirements, and is mainly represented by Quercus robur
L. (pedunculate) and Quercus petraea L. (sessile oak). Bearing
in mind that antioxidant potential of two major Serbian oaksQuercus robur L. and Quercus petraea L. has not been studied
well enough, especially their leaves and twigs, the aim of this
work was to investigate the in vitro antioxidant activities.

Methods: Antioxidant characteristics of leaves, twigs and
acorns from Serbian oak species from Vojvodina province
(northern Serbia) were investigated. 80% EtOH (in water) extracts were used for antiradical power (ARP) determinations
against DPPHo, oNO, O2o- and oOH radicals, ferric reducing
antioxidant power (FRAP), total phenol (TPC) and tannin
(TAC) contents. Permanganate reducing antioxidant capacity
(PRAC) was determined using water extracts. Beside mentioned parameters, soluble proteins, lipid peroxidation (LP),
pigment, proline, flavonoid and proantocyanidine contents
were also determined. The data of different procedures were
compared and analysed by multivariate techniques (correlation
matrix calculation and principal component analysis (PCA)).
Results: Significant positive correlations were obtained between ARP parameters and contents of phenolic antioxidants.
Close interdependence was also observed between PRAC and
O2o- ARP, which were totally opposite to LP on the loading
plot. PCA found that investigated organs of two different oak
tree species possess similar antioxidant characteristics.
Conclusions: The superior antioxidant characteristics
showed oak leaves over twigs and acorns and seems to be promising source of antioxidants with possible use in industry and
pharmacy.
Key words: antioxidant; scavenging capacity; oak tree;
FRAP, DPPH

PO2707
PRODUCTION OF LIPIDS WITH IMPROVED NUTRITIONAL AND BIOLOGICAL PROPERTIES USING
ENZYMATIC INTERESTERIFICATION OF VEGETABLES OILS FROM AMAZONIAN AREA
G. Macedo1, P. Speranza1, A. P. Ribeiro2, J. Macedo3
1
Food Science Department; UNICAMP - Campinas
State University, Campinas, Sao Paulo, Brasil
2

School of Applied Sciences, UNICAMP, Limeira, Sao
Paulo, Brasil
3
Food Nutrition Department, UNICAMP, Campinas,
Sao Paulo, Brasil
Background and objectives: Composition and nature of
oils and fats used in the preparation of food products is acquiring greater importance. It is now known that the incidence of
cardiovascular disease, diabetes and other metabolic syndromes are directly related to the type and amount of lipid consumed. Currently, the search for lipids non-hydrogenated, low
cholesterol and rich in polyunsaturated fatty acids is a need for
the food market. Therefore, oil industries have focused mainly
on upgrading products to better fit the current needs of consumers, who are looking for healthier foods, manufactured with
higher quality. Enzymatic interesterification, involving the rearrangement of fatty acids among glycerol backbones, proving
to be a good alternative. The changes in the original triacylglycerol composition modified the physical and nutritional
properties of reestructured triacylglycerol, thus increasing the
potential applications of these lipids. The objective of this study
was to produce interesterified blends rich in unsaturated fatty
acids using different enzymatic systems.
Methods: For the production of these mixtures were used
buriti and muru-muru oils, natural riches of the Amazon region, with huge unexplored and nutritional potential.
Results: The results obtained of regiospecific distribution of
fatty acids on triacylglycerol (13C NMR) indicate the production of oils rich in unsaturated fatty acids at positions sn-1,3 for
both enzyme systems tested. The concentration of unsaturated
fatty acids at positions sn-1, 3 of triacylglycerol increased from
33.5% in the original mixture to 54.1% in the mixture produced using lipase from Rhizopus sp, and 54.5% in the mixture
produced with a commercially available immobilized lipase.
Conclusions: The production of lipids with high content of
unsaturated fatty acids in the position sn-1,3 of triglyceride and
free of trans fatty acids is of great interest for the development
of lipids with high nutritional and biological potential.
Key words: lipase, amazon oils, interesterification.


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Abstracts


PO2708
METABOLIC TRANSIT OF N-CARBOXYMETHYLLYSINE AFTER CONSUMPTION OF MODEL MAILLARD REACTION PRODUCTS
I. Roncero-Ramos1, C. Delgado-Andrade1, F J. Tessier2,
C. Niquet-Léridon2, C. Strauch3, V. Monnier3, M P. Navarro1
1
Instituto de Nutrición Animal, Estación Experimental del Zaidín, CSIC, Spain
2
Institut Polytechnique LaSalle Beauvais, Beauvais,
France
3
Case Western Research University, Cleveland, Ohio,
USA
Background and objectives: The purpose of this study was
to investigate the intake and excretion of an advanced glycation
compound produced in the Maillard reaction, carboxymethyllysine (CML), after feeding rats a diet containing Maillard reaction products (MRPs) from the glucose-lysine model system.
CML in serum and different tissues was measured to detect
target organs.
Methods: Equimolar mixture of glucose-lysine-HCl (GL)
was heated (150°C, 90 min) to obtain GL90 sample. It was
added to the AIN-93G diet (Control diet) to reach a final concentration of 3% (GL90 diet). Twenty-four weanling rats were
distributed into two groups and assigned to dietary treatments
for three months, monitoring food consumption and body

weights. In the last week, faeces and urine were collected daily
and stored as a 1-week pool. After sacrifice, blood was drawn
to obtain serum, and some organs removed. CML analysis was
performed by HPLC/MS-MS in diets, faeces, urines, serums
and tissues.
Results: CML intake increased after GL90 diet consumption (30 vs. 175µg/day for Control and GL90 group), accordingly these animals excreted much more CML in faeces and
in urine. As percentage of the intake, faecal CML was 2.5-fold
higher in animals fed GL90 diet. However, the rate of urinary
CML was lower in them. No changes were found in proteinbound-CML levels in serum whereas CML in bone, tendon
and heart increased due to MRPs consumption
Conclusions: Faecal CML excretion is highly influenced
by dietary CML levels. The rate of CML excreted in urine was
probably limited or saturated depending on the type of compounds ingested. Consumption of assayed MRPs led to a higher CML deposit in heart and tendon, while in bone the increased amounts of CML ought to have an in situ origin as a
consequence of a dicarbonyl compounds environment.

PO2709
EFFECT OF GARLIC ESSENTIAL OIL ON PROTEIN
OXIDATION IN CHILL STORED PORK PATTIES
G. Nieto1,2, L. Skibsted2, M. Andersen2, G. Ros1
1
University of Murcia, Murcia, Spain
2
University of Copenhagen, Denmark, Spain
Background and objectives: Taking into accounts that
meat products, such as burger patties, are very susceptible to
oxidation as mincing, cooking, and addition of salt promotes
the formation of Reactive oxygen species (ROS), the addition
of natural antioxidants as essential oil on these meat products
could be an interesting strategy to improve their quality. The
aim of this study was to determine the effect of garlic (G) essential oil (EO) on protein oxidation in pork burgers manufactured without synthetic additives and stored under retail display

conditions.
Methods: For that purpose 3 batches of pork burgers (minced to 5 mm and 2 % salt) were prepared: the control group C;
G1 (Level 1: 0.05% EO); and G2 (Level 2: 0.4% EO). The patties were packed with modified atmosphere (70%O2: 20%CO2:
10% N2) (MAP), stored for up to 9 days at 4ºC. The oxidative
stability was evaluated by the formation of myosin cross-links
as analysed by SDS-PAGE, and protein thiol loss as analyzed
spectrophotometrically after derivatisation with DTNB.
Results: Results showed that protein thiols were lost to yield
myosin cross-links in the pork patties during storage.Garlic
was found to promote protein oxidation, as seen by an extreme loss in thiol groups and highly elevated myosin cross-link
formation, indicating that G interacts with the thiol groups of
the myofibrillar proteins, and thus increases the cross-link formation in pork patties stored in oxygen containing packages.
Conclusions: This study shows that essential oils of garlic
should be tested for prooxidant activity before to be used.
Key words:Pork patties; thiol oxidation; protein cross-linking, garlic

Key words: Maillard reaction products; carboxymethyllysine; bone; heart; tendon.

Abstracts

Ann Nutr Metab 2013;63(suppl 1):1–1960

1573


PO2710
ANTIOXIDANT AND PROOXIDANT ACTIVITIES OF
PHENOLIC COMPOUNDS OF THE EXTRACTS OF
ECHINACEA PURPUREA (L.) AND CASTANEA SATIVA MILL
H. Jukic1, I. Mujic2, D. Rudic2, J. Zivkovic3, M. Gregoric4, S.

Kalic5, K. Damjanic2
1
University of Bihac, College of Medical Studies,
Bihac, Bosnia and Herzegovina
2
Collegium Fluminense Polytechnic of Rijeka, Rijeka, Croatia
3
Faculty of Medicine, University of Nis, Department
of Pharmacy, Nis, Serbia
4
Zagreb Health School, Zagreb, Croatia
5
Factory for processing of fruits and vegetables,
Orahovica, Croatia

PO2711
DOCOSAHEXAENOIC ACID DOWN-REGULATES
EGF-INDUCED MATRIX METALLOPROTEINASE-1/-9 AND UPA EXPRESSION VIA DISRUPTION
OF INTERACTION BETWEEN EGFR AND ERBB2
AND INACTIVATION OF EGFR/ERBB-2 SIGNALING
IN SK-BR3 BREAST CANCER CELLS
F. Cheng1, C. Lu1, C. Li12
1
School of Nutrition, Chung Shan Medical University, Taichung, Taiwan
2
Department of Nutrition, Chung Shan Medical
University Hospital, Taichung, Taiwan

Background and objectives: In recent years, there is a
growing interest on natural and safer antioxidants. So far, little

is known about the potential of E. purpurea and C. sativa extracts.
Methods: In order to evaluate its antioxidant activity of extracts investigation had to include the total phenolics content
and the scavenging capacity on DPPHradicals. The ability of
extracts to scavenge superoxide and hydroxyl radicals were tested using electron paramagnetic resonance (EPR) techniques.
Also, the extracts were screened for cytotoxicity using neutral
red chemotherapeutic sensitivity assays on human colon cancer cell lines SW480. The cells were exposed to various concentrations of extracts and different treatment times.
Results: The content of total phenolic compounds of extracts of E. purpurea and C. sativa leaves were 10.57 % GAE
and 6.74 % GAE, respectively. The scavenging activity of the
radicals was found to exhibit 50% of the inhibition value (IC50
value) at the concentration of 15.67 µg/ml for the investigated
of E. purpurea and 1.87 µg/ml for C. sativa leaves. The ability of
C. sativa to scavenge hydroxyl and superoxide radicals was at
the concentration of 42.3 µg/ml and 8.5 µg/ml. Examination of
redox potential E. purpurea on cancer cells SW480 was shown
prooxidant effect at lower concentrations of extract and shorter incubation period. Treatment of cancer cells with chestnut
leaf extract immediately cause a cytotoxic effect with the initial concentration and cytotoxic effect increases as a function
of the concentration of the extract. The highest concentration
was also the most toxic which is particularly evident in the
treatment of 24 hours.
Conclusions: The obrained results shown that investigated extracts have good antioxidant potential. The antioxidant
properties of phenolic compounds C. sativa leaves extracts was
better compared to E. purpurea.

Background and objectives: Matrix metalloproteinase-1
(MMP-1), MMP-9 and urokinase plasminogen activator (uPA)
play crucial roles in tumor metastasis. Previous studies showed
polyunsaturated fatty acids exhibit an anti-cancer effect in various human carcinoma cells; however, the effect of docosahexaenoic acid (DHA; 22:6, n-3) on metastasis of breast cancer
cells is not fully clarified. ErbB2 is overexpressed in 20-30% of
human breast cancers and correlates with poor prognosis.
Methods: We used ErbB2-overexpressing SK-BR3 breast

cancer cells as cell model, and used Western blotting, real-time
PCR, siRNA, and zymography assay to explore the anti-metastasis activity of DHA.
Results: EGF (40 ng/ml) induced MMP-1, MMP-9, and
uPA mRNA and protein expression, as well as enzyme activity in both dose- and time-dependent manners, and 100 £gM
DHA significantly inhibited EGF-induced MMP-1, MMP-9,
and uPA protein expression, enzyme activity, and cell migration. Treatment with SP600125 and PD98059 decreased EGFinduced MMP-1, MMP-9, and uPA protein expression. In
addition, EGF-induced MMP-1 and uPA expression was also
decreased by SB203580 and LY294002DHA was shown to inhibit EGF-induced activation of ERK1/2 and JNK. The phosphorylation of EGFR and ErbB2 was induced within 15 min
after EGF treatment and the total amount of protein was found
to increase after 12 h of EGF exposure. EGF-induced phosphorylation and increased total protein of EGFR and ErbB2
were decreased by DHA. Co-immunoprecipitation analysis
showed the interaction between EGFR and ErbB2 was disrupted by DHA. EGFR siRNA mimics the inhibition of EGF-induced MMP-1, MMP-9, and uPA expression as well as enzyme
activity by DHA.
Conclusions: Taken together, these results suggest attenuation of EGFR and ErbB2 expression and disruption of protein
interaction between EGFR and ErbB2 and their downstream
signaling pathways are involved in DHA’s down-regulation of
EGF-induced MMP-1/-9 and uPA expression in SK-BR3 breast
cancer cells.

Key words: Antioxidants, phenolic compounds, Echinacea
purpurea, Castanea sativa.

Key words: Docosahexaenoic acid; MMP; uPA; EGFR;
ErbB2; SK-BR3

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Ann Nutr Metab 2013;63(suppl 1):1–1960

Abstracts



PO2712
ANTI-OBESITY EFFECTS OF DIETARY D-PSICOSE
IN GROWING RAT FED A HIGH-SUCROSE DIET
M. Ochiai1, K. Onishi1, T. Yamada2, T. Matsuo1
1
Faculty of Agriculture, Kagawa University, Japan
2
Research and Development, Matsutani Chemical
Industry Co., Ltd, Japan
Background and objectives: D-Psicose (D-ribo-2-hexulose), C-3 epimer of D-fructose, is a rare sugar presents in small
quantities in nature product. We previously reported that Dpsicose provided no energy and had anti-obesity effects in rats,
but their detailed mechanisms remain to be elucidated. We
here investigated the effects of anti-obesity effects of dietary
D-psicose in growing rat pair-fed a high-sucrose diet (HSD).
Methods:Thirty-one male Wistar rats aged 4weeks were divided into two dietary groups: HSD contained 5% cellulose (C,
n=21), and HSD contained 5% D-psicose (P, n=10). The rats in
the C dietary group were further divided into two groups: rats
fed the C diet ad libitum (C-AD group, n=11), and rats pairfed the C diet with rats in the P group (C-PF, n=10). Each diet
was fed for 8 week. At the weeks between 5th and 7th, energy
expenditure (EE) was measured. At the end of the test period,
all rats were killed by decapitation after 4-hours fasting. The
serum and tissues were collected for biochemical analysis.
Results:The food intake was significantly lower in the P
and C-PF groups than in the C-AD group. The body weight
gain and food efficiency did not differ between the P and C-PF
groups. The EE in darkness, lipoprotein lipase activity of the
soleus muscle, and hepatic glycogen content were significantly
higher, and in contrast, the serum levels of glucose, leptin, and

adiponectin, glucose 6 phosphate dehydrogenase activity, body
fat accumulation were significantly lower in the P group than
in the C-PF group.
Conclusions:The anti-obesity effects of D-psicose could be
induced by not only the suppression of food intake, but the increases of EE in rats.
Key words: D-psicose, pair-feeding, obesity, energy expenditure, rat

PO2713
ASPERGILLUS AWAMORI-FERMENTED BURDOCK
ELEVATES BIFIDOBACTERIUM AND REDUCES
ADIPOSE TISSUE WEIGHT IN RATS FED A HIGHFAT DIET
V N. Sitanggang1, S. Sato1, N. Ohnishi1, J. Inoue2, T. Iguchi3, T. Watanabe3, Y. Okazaki4, H. Tomotake5, N. Kato1
1
Graduate School of Biosphere Science,Hiroshima
University, Japan
2
Ahjikan Co. Ltd., Japan
3
Yaegaki Bio-industry Incorporated,Japan
4
Faculty of Human Life Sciences, Fuji Women’s University, Japan
5
Iida Women’s Junior College,Japan
Background and objectives: Aspergillus has been used widely for the production of sake, miso, soy source, etc. in Japan.
However, there is very limited study on the utilization of Aspergillus for the production of functional foods. Burdock, Artium lappa, is a popular vegetable in Japan, and has been used
therapeutically in Asia, Europe, and North America for hundreds of years. Burdock contains high amounts of dietary fibers such as inulin and oligosaccharides as well as polyphenols
such as chlorogenic and caffeic acid. Recently, to improve the
functional properties of burdock, we have developed a method
for producing Aspergillus awamori-fermented burdock. This
study was conducted to investigate the effects of dietary supplementation with the fermented burdock powder on colonic luminal environment and body fat in rats fed a high-fat diet.

Methods: Male SD rats (5 weeks old) were fed on a high-fat
diet (30% beef tallow) containing 5% burdock powder or 5%
Aspergillus awamori-fermented burdock powder for 3 weeks.
Results: Food intake and growth were unaffected by dietary
manipulation. Consumption of the burdock and fermented
burdock diets significantly elevated fecal IgA and mucins (indices of intestinal immune and barrier functions) and reduced
fecal lithocholic acid (a risk factor for colon cancer) (p< 0.05).
Interestingly, the fermented burdock diet markedly elevated
cecal Bifidobacterium and organic acids including lactate, acetate, propionate and butyrate, and reduced fecal deoxycholic
acid (a risk factor for colon cancer) and perirenal adipose tissue weight (p< 0.05), but the burdock diet did not.
Conclusions: The results suggest that the consumption of
fermented burdock improves the colonic luminal environment
and suppresses obesity in rats fed a high-fat diet.
Key words: burdock, Aspergillus awamori, Bifidobacterium, obesity, secondary bile acids

Abstracts

Ann Nutr Metab 2013;63(suppl 1):1–1960

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