## QUALITY INDICATOR IN THE NUTRITIONAL CARE OF CRITICALLY ILL PATIENTS IN A PRIVATE HOSPITAL - BRAZIL

(1)Konkolisc Pina de Andrade, Grasiela; (2)Chaloub Chediac Rodrigues, Ana Lucia; (2)Silva, Luanye Karla; (3)Nadolskis Severine, Ariane; (4)Junqueira Amarante, Gustavo Adolpho.

(1)Nutritionist. Division of Nutrition. Hospital Sírio Libanês. Brazil;

(2)Nutritionist. Division of Nutrition. Hospital Sírio Libanes. Brazil;

(3)Msc. Nutritionist. Division of Nutrition. Hospital Sírio Libanes. Brazil; (4)MBA. Doctor. Division of Critical Care Medicine.

**Background and objectives:** Patients on nutritional therapy should be monitored routinely, and this evaluation should ensure that the best therapy is being offered for clinical recovery. Our objective was to create a tool to measure and monitor the quality of therapy provided to the critical patient aiming at nutritional recovery.

**Methods:** All hospitalized patients underwent a nutritional evaluation and were classified into 2 groups: with or without nutritional risk. Patients with nutritional risk were reevaluated every 7 to 10 days, taking into account: tolerance of the oral, enteral or parenteral therapy, biochemical tests and anthropometric measures. Based on the results, the patient was classified into three categories: worsening, maintenance or improvement of nutritional status. The monthly goal was to achieve at least 75% maintenance or improvement of nutritional status in these patients.

**Results:** The proposed target was reached between January and June 2016 (mean 92.1%), so in July a new target of 92% was proposed. Between July 2016 and April 2017, only 2 monthly measurements showed results below the expected target: October 2016 (91.5%) and January 2017 (89.3%). Analyzing the data, specific actions were implemented to reverse the critical points of nutritional care like early oral supplementation or enteral therapy.

**Conclusions:** Nutritional therapy is essential for the critical patient and the implementation of tools to measure the quality of care contributes to an effective and individualized nutritional therapy aiming at the clinical recovery of patients, at a lower cost.

**Keywords:** Quality Indicator, Nutrition Assistance

## BIOMARKERS OF INFLAMMATION AND MICRO-NUTRIENT CONCENTRATIONS IN RESPONSE TO A NOROVIRUS IMMUNOLOGIC CHALLENGE

(1)Williams, Anne; (2)Leon, Juan S.; (3)Lopman, Benjamin A.;

(4)Tangpricha, Vin; (5)Whitehead, Jr., Ralph D.; (6)Flores-Ayala, Rafael;

(7)Suchdev, Parminder S.

(1)PhD. Centers for Disease Control and Prevention; (2)PhD, MPH. Hubert Department of Global Health. Emory University. USA.; (3)PhD. Department of Epidemiology. Emory University. USA.; (4)MD, PhD. Department of Medicine. Emory University School of Medicine. USA.; (5)PhD. International Micronutrient Malnutrition Prevention and Control. Centers for Disease Control and Prevention. USA.; (6)DrPH. Nutrition Branch. Centers for Disease Control and Prevention. USA.; (7)MD, MPH. Department of Pediatrics. Emory University School of Medicine.

**Background and objectives:** Characterizing the effects of inflammation and the acute phase response on nutrient biomarkers is a critical research gap to improve accurate assessment of micronutrient status. The objective of this study was to quantify the relationship between Norovirus infection and common biomarkers used to assess inflammation [C-reactive protein (CRP) and alpha-1-acid glycoprotein (AGP)] and micronutrient status.

**Methods:** A longitudinal norovirus challenge study was conducted among apparently healthy adults at Emory University from 2006-2009. Of 64 volunteers, 26 became infected (norovirus detectable in stool by RT-PCR). Sandwich ELISA technique was used to measure proteins in banked serum collected at ten time points including one pre-challenge and nine post-challenge samples (1,2,3,4,7,14,21,28 and 35d post-exposure). A repeated measures study of 52 subjects (age, gender matched) was conducted to assess change in CRP, AGP, ferritin, transferrin receptor (sTfR), and retinol binding protein (RBP) over time by norovirus infection status. We calculated range of biomarker concentrations across the 35d period and report median range biomarker values. The non-parametric rank median test was used to assess differences in range of concentrations over time by infection status.

**Results:** Among infected individuals, CRP, AGP and ferritin concentrations rose to peak values by day 3-4 post-exposure, while uninfected individuals did not show increased concentrations post-exposure. RBP concentrations of infected individuals declined up to day 4-7 post-exposure. Median (quartile1, quartile3) range of biomarker concentrations [uninfected; infected] over time were CRP(mg/L) [1.8 (0.8, 3.2); 17.0 (7.8, 29.3) (p<0.001)], AGP(g/L) [0.4 (0.2, 0.5); 0.6 (0.5, 0.8) (p=0.001)], ferritin (µg/L) [30.7 (15.8, 46.3); 58.1 (48.0, 85.3) (p=0.001)], sTfR (mg/L) [1.5 (1.2, 3.1); 1.8 (1.4, 2.4) (p=0.27)], and RBP(µmol/L) [0.7 (0.4, 0.9); 0.8 (0.6, 1.2) (p=0.10)].

**Conclusions:** The median concentrations ranges of CRP, AGP, and ferritin were greater among infected than uninfected volunteers. Quantifying the magnitude and duration of micronutrient concentration changes, over the course of norovirus infection, will improve the understanding of micronutrient status in areas with high prevalence of inflammation and common infection from gastroenteritis.
**Keywords:** Inflammation, Acute Phase Response, Norovirus, Vitamin A, Iron

**Conflict of Interest Disclosure:** None of the authors declare a conflict of interest. The findings and conclusions in this abstract are those of the authors and do not necessarily represent the official position of the Centers for Disease Control and Prevention.

**Further collaborators:**

This work is supported by the Robert W. Woodruff Foundation at Emory University.

144/2985

## AGREEMENT BETWEEN TWO NUTRITIONAL ASSESSMENT METHODS FOR PREGNANT ADOLESCENTS WHO ATTEND PUBLIC HEALTH SERVICES OF PARAGUAY

(1)Masi, Celia; (2)Gonzalez Cañete, Natalia Elizabeth; (3)Sánchez, Susana; (4)Zaracho de Irazusta, Juana Concepcion; (5)Mendoza de Arbo, Laura.

(1)MSc. Masi Celia. MSc. González-Cañete Natalia. MD Zaracho Juana. MSc. Sánchez S. MSc. Mendoza de Arbo Laura. Instituto Nacional de Alimentación y Nutrición-INAN. Ministerio de Salud Pública y Bienestar Social. Paraguay; (2)MSc. González-Cañete, Nutritionist. Instituto Nacional de Alimentación y Nutrición-INAN. Ministerio de Salud Pública y Bienestar Social. Paraguay;

(3)Msc. Sanchez Susana. Pediatrician. Instituto Nacional de Alimentación y Nutrición-INAN. Ministerio de Salud Pública y Bienestar Social. Paraguay; (4)MD. Zaracho Juana. Pediatrician. Instituto Nacional de Alimentación y Nutrición-INAN. Ministerio de Salud Pública y Bienestar Social. Paraguay.

**Background and objectives:** Introduction: Nutritional assessment of adolescent pregnant women is vital to ensure maternal and fetal health. Most countries in Latin America use Atalah et al. charts to assess nutritional status of pregnant women. In Paraguay Rosso-Mardones criteria is used. Objectives: to evaluate the agreement between two nutritional assessment methods for pregnant women, Rosso-Mardones and Atalah et al.

**Methods:** Material and Methods: analytical cross sectional study. Analysis of sociodemographic and anthropometric data of pregnant adolescents collected in 2015 from the Food and Nutrition Surveillance System (SISVAN by its Spanish acronyms) at public health services of 18 Sanitary Regions. Nutritional status was assessed by Rosso-Mardones and Atalah et al criteria. The results were compared using percentages and the concordance between both methods was determined by Cohen's Kappa. A p value <0,05 was considered significant.

**Results:** 2.208 pregnant adolescents between 15 and 19 years old were included. The average age was 17, 4±1,3 years old; 58,2% lived in urban areas (n=1284) and 78% were in a couple relationship. Nutritional status according to Rosso- Mardones vs Atalah et al.

al. criteria was: underweight 43, 8% (n=966) vs 34% (n=750), normal weight 31,5% (n= 696) vs 46,6% (n=1.028), overweight 10,2% (n=226) vs 14,1% (n=311) and obesity 14,5% (n=320) vs 5,4% (n=119). Cohen's Kappa indicated a good agreement between both methods: 96,4% for underweight, 65,2% for normal weight, 38,3% for overweight, and 100% for obesity(0,617; p=0,0001).

**Conclusions:** Even though underweight and obesity were higher according to Rosso-Mardones, a good concordance was observed between both methods. At the national level there is no consensus on the best criterion for nutritional assessment of pregnant adolescents. Therefore, it is essential to analyze the epidemiological situation of this population and to establish the objectives to be achieved at the public health level in order to select the most appropriate method.

**Keywords:** Nutrition Assessment, nutritional status, pregnancy in adolescence.

144/3005

## COMPARATIVE STUDY OF THE AMINO ACID COMPOSITION OF PEELS, PULP AND CORE OF SEEDLESS BREADFRUIT (ARTOCARPUS ALTILIS)

Esekheigbe, Ayemhenre.

PhD. Senior Lecturer. Ambrose Alli University. Ekpoma. Nigeria.

**Background and objectives:** Seedless breadfruit (Artocarpus altilis) is a large, round, starchy fruit which is usually eaten either boiled or fried. Originally native to the West Indies, it has found its way to Africa including Nigeria. The pulp of the seedless breadfruit which is generally consumed is known to be rich in nutrients notably carbohydrates. However, there is no much information on the nutrient content of the parts not commonly consumed hence the need for the study.

**Methods:** Samples of mature seedless breadfruit were purchased from the market at Ekpoma (6.7491oN, 6.0732E), Edo State, Nigeria. In the laboratory, the samples were peeled and the pulp separated from the core. Thereafter, they were pulverized and dried in an oven at 60oC for 48 hours before being milled. They were then stored in air tight containers at 4oC in the refrigerator before analysis. Protein (N x 6.25) was determined by the Kjedahl method while the amino acid profile in the different seedless breadfruit samples were determined using the ion exchange chromatographic method (Technicon sequential multi-sample amino acid analyzer).

**Results:** From the results obtained, breadfruit was found to contain both essential and non essential amino acids. The total amino acid content was highest in the core, followed by the pulp and peel with values of 714.1, 617.7 and 589.0 mg/g protein respectively. Interestingly however, although the core was found to have the highest concentration of amino acids, the peel which is usually discarded was found to contain the highest percentage of essential amino acids (48.64%) This was followed by the core (45.97%) and the pulp (44.83%). Every portion of the seedless breadfruit is thus important with respect to amino acid content.
**Conclusions:** There is therefore the need to harness the essen-
tial amino acids in the peel of the seedless breadfruit for both man
and animal consumption.

**Keywords:** Amino acid, Breadfruit

**Further collaborators:** Osagie Victor

144/3010

ASSOCIATION OF IODINE STATUS WITH SALT INTAKE AND SALT IODINE CONTENT IN SCHOOL CHILDREN AND ADOLESCENTS

(1)Sánchez Bernal, Susana; (2)Acosta, José; (2)Morínigo, Geraldine;

(3)Masi, Celia; (4)Redondo, Juana; (5)Vega, Gloria; (6)Arias, María

Nidia; (7)Mendoza de Arbo, Laura; (8)Galeano, Marcelo; (9)Zaracho de

Irazusta, Juana Concepcion.

(1)Master. MD pediatrician. National Food and Nutrition Institute.
Ministry of Public Health and Social Welfare. Asunción, Paraguay;

(2)Nutritionist. National Food and Nutrition Institute. Ministry of
Public Health and Social Welfare. Asunción, Paraguay; (3)Master.

Nutritionist. National Food and Nutriton Institute. Ministry of
Public Health and Social Welfare. Asunción, Paraguay; (4)Nurse.

National Food and Nutriton Institute. Ministry of Public Health
and Social Welfare. Asunción, Paraguay; (5)Biochemistry. National
Food and Nutriton Institute. Ministry of Public Health and Social
Welfare. Asunción, Paraguay; (6)Food Technologist. National

Food and Nutriton Institute. Ministry of Public Health and Social
Welfare. Asunción, Paraguay; (7)Master. Physician. National Food
and Nutriton Institute. Ministry of Public Health and Social

Welfare. Asunción, Paraguay; (8)Nutritionist. National Food and
Nutriton Institute. Ministry of Public Health and Social Welfare.
Asunción, Paraguay; (9)Pediatrician. National Food and

Nutriton Institute. Ministry of Public Health and Social Welfare. Asunción, Paraguay.

**Background and objectives:** Salt fortification with iodine is mandatory to prevent, control and eradicate iodine deficiency disorders. It is important to monitor iodine consumption and its content in salt, but it is also necessary to prevent the excess of its intake.

The aim was to evaluate the association between iodine sta-
tus in children and adolescents, with sodium intake through its
urinary levels and with the content of iodine in salt available at
household level.

**Methods:** Cross -sectional, descriptive and analytic study. Previous written consent, urinary iodine was dosed in children and adolescents (ammonium persulfate method) and evaluated for status according to WHO/VMNIS (Vitamin and Mineral Nutrition Information System) criteria. Sodium (Na) was also measured in early spot urine samples by the Ion-Selective Electrode (ISE) method as an indicator of salt consumption. Iodine levels were determined in salt samples (volumetric method-Thiosulfate). ANOVA and Chi square Test was used, p value <0.05 was considered significant.

**Results:** old); 53,1% were woman. The median urinary iodine level was 301.2 mg/L (51-646 mg/L), according iodine status 4,8% had mild iodine deficiency (50-99mg/L), 16.4% was adequate (100-199mg/L), 32.1% was more than adequate (200-299mg/L), 46.7% shows risk of adverse health consequences (>300mg/L). Average urinary Na was 161.2 mmol/L (9.0-459.2mmol/L), it was lower for children with mild iodine deficiency (113.6mg/L) versus those with risk of adverse health consequences (170.5 mg/L), more than adequate (168.1 mg/L) or adequate (147.2mg/L) iodine status (ANOVA, p<0.0001). Iodine levels were adequate (20-40 mg/k) in 60% of salt samples, insufficient (<20 mg/k) in 14.8% and excessive (>40mg/k) in 25.2%, none was negative. The percentage of risk of adverse health consequences of iodine status was 29% when iodine content in salt was excessive vs 20% when salt iodine was adequate (c2, p=0,02). Correlation between urinary Na and iodine was no found (Pearson correlation r=0,145).

**Conclusions:** By iodine urinary, 1 of each 2 child/adolescent present iodine status of risk of adverse health consequences, which is associated with higher intake of salt and when salt con- tains iodine above the recommended.

**Keywords:** iodine, salt, iodine urinary, children, adolescents

**Further collaborators:**

Julia Coronel. Nutritionist. National Food and Nutrition In-
stitute. Ministry of Public Health and Social Welfare. Asunción,
Paraguay

Paolo Barreto. Nutritionist. National Food and Nutrition In-
stitute. Ministry of Public Health and Social Welfare. Asunción,
Paraguay

144/3410

USE OF DRIED MATRIX SPOTS FOR NUTRITION-
AL BIOMARKER ANALYSIS

(1)Arredondo, Francisco X.; (2)Craft, Neal E.; (3)Fleshman, Matthew K.

(1)MD/PhD. Biomarkers Group. Craft Technologies, Inc. USA;

(2)PhD. President. Craft Technologies, Inc. USA.; (3)PhD. Mass Spectrometry Group. Craft Technologies, Inc. USA.

**Background and objectives:** Background

Dry Matrix Spots (DMS) are dried spots of fluids, including
blood, serum, urine, saliva and breast milk, which have been used
for remote collection. A device separating RBCs from plasma
without centrifugation would be useful for nutritional assessment
in developing countries.

Objectives

To obtain and evaluate blood collection devices (BCDs) for
measurement of nutritional biomarkers (NBs). Using DMS, to
determine the best methods to measure NBs, including: ferritin,
folate, retinol binding protein (RBP), 25OH-D, B12, thyroglobu-
lin (Tg), zinc, methylmalonic acid (MMA), homocysteine (HCY),
alpha-1-acidglycoprotein (AGP), and C-reactive protein (CRP).

**Methods:** Criteria were established for successful BCDs and analytical methods. Nine BCDs were obtained and tested. Analytical methods included single-plex enzyme immunosorbent assays
(EIA), microbiological assays (folate, B12), chemiluminescence multiplex EIA, and LC-MS/MS (25OH-D, MMA and HCY).

Short-term folate stability was tested in the ViveBio-collected RBCs using a microbiological assay. Both dry cells and liquid-stabilized were tested at 23°C, 37°C and 45°C. RBC volume was calculated using the measurement of potassium.

**Results:** None of the BCDs met the selection criteria. Many exhibit chromatographic effects, provide too little plasma for analytical testing, or do not separate RBCs from plasma.

A prototype BCD from ViveBio (Alpharetta, GA) that separates RBCs in a vertical format yielding hemoglobin-free plasma on a collection pad was used, permitting analysis of serum-based components and RBC markers.

RBC folate was most stable in the dry cells. A cocktail containing sodium azide and bovine serum albumin (BSA) provided the best results of the liquid stabilizers.

Some NBs could not be measured using single-plex EIAs due to sample dilution, therefore, the QuanSys multiplex was used successfully. It offers the advantage of obtaining results for 6 NBs (AGP, CRP, Ferritin, Tg, RBP, and TfR) simultaneously.

Using LC-MS/MS, MMA, HCY and 25OH-D were successfully measured in eluates from DMS and in extracts stored at 23°C, 37°C and 45°C.

**Conclusions:** ViveBio DMS collection device was identified which separates RBCs from plasma. Both compartments were used to test NBs. The stability of 12 NBs at 5 temperatures is being measured in DMS using QuanSys multiplex, microbiological, and LC-MS/MS methods.

**Keywords:** biomarkers, nutrition assessment, dry blood spots, vitamins, multiplex

**Further collaborators:** Funded by the Bill and Melinda Gates Foundation

(¹)Eldridge, Alison; (²)Cade, Janet; (³)Gibney, Mike; (⁴)Gurinovic, Mirjana; (⁵)Illner, Anne-Kathrin; (⁶)Piernas, Carmen; (⁷)Smith, Benjamin; (⁸)Taberner, Jonathon; (⁹)de Vries, Jeanne.

(¹)PhD. Institute of Nutritional Science. Nestlé Research Center. Switzerland.; (²)PhD. School of Food Science and Nutrition. University of Leeds. United Kingdom.; (³)PhD. Institute of Food and Health. University College Dublin. Ireland.; (⁴)PhD. Centre of Research Excellence in Nutrition and Metabolism. Institute for Medical Research. University of Belgrade. Serbia.; (⁵)PhD MPH. Department of Nutrition and Health Sciences. Polytechnic Institute UniLaSalle Beauvais. France.; (⁶)PhD. Nuffield Department of Primary Care Health Sciences. University of Oxford. United Kingdom.; (⁷)PhD. Toxicology and Scientific Services. Firmenich. United States of America.; (⁸)Student. School of Food Science and Nutrition. University of Leeds. United Kingdom.; (⁹)PhD. Department of Agrotechnology and Food Sciences. Wageningen University. The Netherlands.

**Background and objectives:** There is growing pressure in the area of dietary intake assessment to improve the accuracy and reduce costs of data collection and processing. New technology-based tools are available and more are under development, including web-based programs, mobile applications, and digital image-assisted and digital image-based tools, among others. The ILSI Europe Dietary Intake and Exposure Task Force launched this project to characterize and evaluate technology-based dietary assessment tools in order to develop general quality standards for future applications.

**Methods:** A comprehensive literature review identified new technology-based tools using key word searches with the following inclusion criteria: publications were in English, papers were published from 2011-2016, and the tool features, functions and uses were detailed. A total of 29 dietary assessment tools were identified. Scoring criteria were developed to evaluate tool features, results outputs, sources and completeness of food composition data, target audience and suitability for research. Each tool was rated on 27 features.

**Results:** Most of the tools identified (23/29) relied on self-reporting of dietary intake data, either through web-based programs or mobile apps. Eleven used digital images to help identify and quantify foods consumed, and one used a barcode scanner. Most tools reported energy (22/29) and macronutrients (21/29), but fewer reported micronutrients (19/29) and food groups (17/29). Only 16/29 allowed for data to be exported to researchers or other health professionals. Few publications on the tools included details about food composition tables or the ability to add missing items (5/29) or custom recipes (2/29). Scores for the tools ranged from 4 to 18 with an average of 11 out of 27 possible attributes.

**Conclusions:** Dietary assessment methods that utilize technology provide rapid feedback to users and offer potential cost-savings for researchers. There remain gaps in many of these tools before they will be ready to replace more traditional interview-based
methods for research purposes, and most require validity testing, additional description of the food composition tables used and details on how the foods are identified and quantified. This project will provide perspective on quality standards that could be recommended for future development and reporting of technologies in the area of dietary intake assessment.

**Keywords:** Dietary assessment, digital images, mobile app, web-based tools.

# Track 6: Functional Foods and Bioactive Compounds

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## EFFECTS ON POSTPRANDIAL BLOOD GLUCOSE AND INSULIN LEVELS AND HYDROGEN EXCRETION THROUGH THE INHIBITORY EFFECTS OF 1, 5-ANHYDROGLUCITOL ON DISACCHARIDASES IN RATS AND HEALTHY HUMANS

**Background and objectives:** 1,5-Anhydro-D-glucitol (1,5-AG) is found in low levels in natural plant foods and also produced sparingly from glycogen in the liver. 1,5-AG which is mostly contained in the body is used as a marker of blood glucose control in patients with diabetes mellitus. The objective is to investigate whether 1,5-AG, which is ingested excessively as a food ingredient, has a specific physiological function.

**Methods:** The inhibition by 1,5-AG was determined on disaccharidases of small intestine of rats and humans, and compared with that of rare sugars. To investigate the metabolism and fate of 1,5-AG, 1,5-AG was administered orally to rats prior to human experiments and the effects on blood concentrations and urinary excretion of 1,5-AG and hydrogen excretion were determined. In addition, a sucrose or glucose solution with or without 1,5-AG was administered orally to rats, and the suppressive effects on postprandial blood glucose and insulin was determined. Similar physiological functions were also investigated using healthy human participants.

**Results:** 1,5-AG distinctly inhibited the activity of sucrase, maltase, trehalase and lactase, while D-sorbose and L-arabinose strongly inhibited sucrase but barely inhibited trehalase and lactase. The inhibition of 1,5-AG on sucrase and maltase were similar between humans and rats. 1,5-AG in serum showed a peak 30 min after ingestion of 1,5-AG (10 g) by healthy subjects, and decreased gradually over 180 min. About 60% of 1,5-AG was excreted into the urine for 9 h following ingestion, while mostly 100% of 1,5-AG was excreted into the urine 24 h after administration in rats. Hydrogen was scarcely excreted in both rats and humans 24 h after administration of 1,5-AG. 1,5-AG significantly suppressed the elevation of blood glucose and insulin by the ingestion of sucrose or glucose. A large amount of 1,5-AG (20 g) caused no side effects among healthy participants. Ingested 1,5-AG was readily absorbed from the small intestine and excreted rapidly into the urine. Thus, the available energy of 1,5-AG is 0 kcal/g and 1,5-AG does not cause osmotic diarrhoea.

<footnote>(1)Oku, Tsuneyuki; (2)Tanabe, Kenichi; (3)Yoshinaga, Kazuhiro;</footnote>

<footnote>(4)Shimura, Fumio; (4)Nakamura, Sadako.</footnote>

<footnote>(1)Jumonji University; (2)PhD. Department of Food Science and Nutrition. Nagoya Women's University. Japan.; (3)PhD. Developing Laboratory of New Materials. Nihon Starch Co. Ltd. Japan.; (4)PhD. Institute of Food, Nutrition & Health. Jumonji University. Japan.</footnote>
**Conclusions:** 1,5-AG which has a very low toxicity and is safe, could be use as low energy bulking sweetener in food products.

**Keywords:** 1,5-anhydroglucitol; postprandial blood glucose; disaccharidase; hydrogen excretion; inhibitory effect.

144/248

EFFECTS OF 1,5-ANHYDROGLUCITOL-FEEDING ON GROWTH, ORGAN WEIGHT, BLOOD BIO-CHEMICAL MARKERS AND OXIDATIVE STRESS IN RATS

**Background and objectives:** 1,5-anhydro-D-glucitol (1,5-AG) which has 60 % of the sweetness of sucrose, is found in low levels in natural plant foods and also produced sparingly from glycogen in the liver. 1,5-AG which is mostly contained in the body, is used as a marker of blood glucose control in patients with diabetes mellitus. The objective is to investigate whether 1,5-AG, which is ingested excessively as a food ingredient, does not cause the harmful effect and has a specific physiological function.

**Methods:** Male Wistar rats aged 4-wk were raised using consecutive feeding of diets (AIN93G) containing 5% and 10% of 1,5-AG for 31 days (n=5 in each group). Erythritol, which is easily absorbable and excreted in the urine without degradation, and fructooligosaccharide (FOS), which is non digestible and completely fermentable were used as controls. Body weight and food intake were determined every day and blood was collected on 24th day of experiment to measure oxidative stress. The urine and feces were collected between 27th and 29th day of experiment using metabolic cage.

**Results:** The growth and organ weights of the rats were not significantly different between the control group and the other 4 groups, because the amount of dietary intake was adjusted to become identical by light restricted feeding. In the experiment of revenue and expenditure, almost 100% of the 1,5-AG ingested was recovered from urine collected for 24 h. These results demonstrate that almost 1,5-AG ingested is absorbed from the small intestine and is excreted in the urine without degradation, with the available energy being 0 kcal/g. 1,5-AG did not affect the concentration of blood glucose or triacylglycerol following consecutive feeding for 31 days and also did not induce morphological abnormalities in organs and tissues without cecal enlargement in the FOS group. The anti-oxidative potential and oxidative index of plasma and the

urinary excretion of 8-hydroxy-2'-deoxyguanosine were not significantly different between the five groups.

**Conclusions:** Ingestion of 1.6 g of 1,5-AG/kg body weight per day (average intake during experiment) does not have harmful effects on the growth and physical condition of rats. Taken together these results demonstrate that 1,5-AG is a safe ingredient in food.

**Keywords:** 1,5-anhydroglucitol; oxidative Stress; blood biochemical markers; organ weight.

144/254

ACCEPTABILITY OF FORTIFIED WHEY PROTEIN CONCENTRATE SUPPLEMENTS WHEN ADDED TO CUSTOMARY THIN GRUELS (ATOLES) BY WOMEN IN THE WESTERN HIGHLANDS OF GUATEMALA

**Background and objectives:** Dietary surveys have shown a gap in the daily intake of protein and other micronutrients, among lactating women in the western highlands of Guatemala. Culturally, during lactation, women consume additional amounts of thin gruel beverages (atoles), which can serve as a vehicle to secure the missing nutrients. Simple addition of whey-protein concentrate (WPC) supplement to thin gruels has been found acceptable; nonetheless addition of micronutrients could change some organoleptic characteristics. We sought to determine, among women of different socio-demographic settings, the acceptability of fortified WPC supplements added to thin gruels.

**Methods:** 55 women aged 17-70y were recruited locally across two socio-demographic settings in Quetzaltenango, Guatemala. They were asked to bring a home-prepared gruel of their preference.

Three cups containing 100 ml of the gruel and different WPC presentation were served for sensory evaluation: alone; fortified with recommended dosage of micronutrients for lactation (FWPC); and fortified with twice the recommended dosage of micronutrients for lactation (2xFWPC). Sensory evaluation involved responding as to any detectable differences in taste and selecting their most and least preferred options.

**Results:** 91% of the women detected a difference in taste among the beverages. Participants were equally inclined to choose either WPC or 2xFWPC as their preferred beverage, both reported at 38%, with 24% favoring FWPC. For the least preferred beverage,

<footnote>(1)Oku, Tsuneyuki; (2)Nakamura, Sadako; (3)Tanabe, Kenichi;</footnote>

<footnote>(4)Miyahara, Akemi; (5)Izumi, Chiyoko; (6)Yoshinaga, Kazuhiro;</footnote>

<footnote>(7)Yamazaki, Yuko; (2)Sasaki, Naho.</footnote>

<footnote>(1)Jumonji University; (2)PhD. Institute of Food, Nutrition & Health. Jumonji University. Japan.; (3)PhD. Department of Food Science and Nutrition. Nagoya Women's University. Japan.;</footnote>

<footnote>(4)BSc. Graduate School of Human Health Science. University of Nagasaki. Japan.; (5)MSc. Developing Laboratory of New Materials. Nihon Starch Co. Ltd. Japan.; (6)PhD. Developing Laboratory of New Materials. Nihon Starch Co. Ltd. Japan.; (7)PhD. Institute of Food, Nutrition & Health. Jumonji University. Japan.</footnote>

<footnote>(1)Arriaga, Claudia Lorena; (2)Fuentes, Deborah; (2)Diaz-Ruiz, Elena;</footnote>

<footnote>(3)Garcia-Meza, Rosario; (4)Solomons, Noel W; (5)Hirst, Daniel;</footnote>

<footnote>(6)Bonorden, Melissa.</footnote>

<footnote>(1)Researcher. Center for Studies of Sensory Impairment, Aging and Metabolism.; (2)Nutritionist. Center for Studies of Sensory Impairment, Aging and Metabolism.; (3)MA. Nutritionist. Center for Studies of Sensory Impairment, Aging and Metabolism.;</footnote>

<footnote>(4)Scientific Director. Center for Studies of Sensory Impairment, Aging and Metabolism.; (5)MSc. Food Scientist - Product Development Hormel Foods Corporation; (6)MSc. Senior Food Scientist - Product Development Hormel Foods Corporation.</footnote>
44% chose 2xFWPC and WPC and FWPC tied with 28% each. When responding to the question of voluntarily drinking either of the fortified offerings on a regular basis, 97% of the women responded that they would do so willingly.

**Conclusions:** A fortified WPC supplement, mixed with thin traditional gruels, proves to be not only a culturally-pertinent, but also a plausible vehicle for the micronutrients needed to fill the existing gaps to reach recommended intakes for lactation.

**Keywords:** Acceptability, whey protein concentrate, thin gruel beverages, guatemalan western highlands

144/268

COMPUTER-AIDED TOOLS IN THE ANALYSIS OF TASTANT/BIOACTIVE PEPTIDES OCCURRING IN FOOD-ORIGINATING PROTEIN SEQUENCES

(1)Iwaniak, Anna; (2)Darewicz, Malgorzata; (3)WOŁOWSKA, Ewelina;

(2)Minkiewicz, Piotr.

(1)Professor.Warmia and Mazury University in Olsztyn. Faculty of Food Science. Chair of Food Biochemistry. Olsztyn. Poland;

(2)Professor. Warmia and Mazury University in Olsztyn. Faculty of Food Science. Chair of Food Biochemistry. Olsztyn. Poland;

(3)MSc. Warmia and Mazury University in Olsztyn. Faculty of Food Science. Chair of Food Biochemistry. Olsztyn. Poland.

**Background and objectives:** Peptides significantly affect the sensory attractiveness of foods, because they feature the taste properties of products by affecting the following taste sensations: umami, sour, sweet, salty and bitter. The latter one can be regarded by consumers as undesired, especially when thinking about ripening cheeses and formulas composed of protein hydrolysates.

**Methods:** The aim of the study was the computer analysis of food derived protein sequences being potential source of tastant peptides. Analyses included: elaboration of the profiles of potential sensory activity of proteins as well as the computer simulation of hydrolysis of proteins to generate tastant/bioactive peptides. The selected plant and animal protein sequences (derived from oil and leguminous plants, crops, milk, fish, and meat) were taken for the bioinformatic studies. **Methods:** **Results:** Based on the profile of potential sensory activity of proteins, we found that majority of peptides located in proteins were bitter. Moreover, potentially the largest number of tastant peptides was present in milk and carp proteins (caseins and myosin, respectively). The data concerning the milk proteins as the source of bitter peptides were consistent which the results confirmed in in vitro studies. Many peptides located in proteins were dipeptides. It was found that ficin and papain were the proteases, which potentially released the largest number of taste peptides from all protein sequences analyzed. It was also concluded that some peptide tastants revealed biological potential as the dietary components supporting the metabolic syndrome prophylaxis. Our studies confirmed the suitability of selected bioinformatic tools in studying the proteins as the food components being the source of tastant/bioactive peptides.

**Conclusions:** Our studies confirmed the suitability of selected bioinformatic tools in studying the proteins as the food components being the source of tastant/bioactive peptides.

**Keywords:** sensory peptides, proteins, in silico analysis, hydrolysis

**Further collaborators:**

Work supported from Warmia and Mazury University funds.

144/271

DIFFERENTIAL PERIPHERAL BLOOD METHYLATION BY ALPHA-LIPOIC ACID AND EPA SUPPLEMENTATION IN OVERWEIGHT/OBESE WOMEN DURING A WEIGHT LOSS PROGRAM

(1)Huerta Hernández, Ana Elsa; (1)Riezu Boj, José Ignacio; (1)Milagro Yoldi, Fermín Ignacio; (2)Guruceaga Martínez, Elizabeth; (1)Moreno Aliaga, María Jesús; (3)Martínez, J. Alfredo.

(1)PhD. Department of Nutrition, Food Science and Physiology. University of Navarra. Spain.; (2)MsC. Proteomics & Bioinformatics Unit. Center for Applied Medical Research (CIMA). Spain.; (3)MD. PhD. Department of Nutrition, Food Science and Physiology. University of Navarra. Spain.

**Background and objectives:** It has been reported that α-lipoic acid (LA) supplementation alone or in combination with eicosapentaenoic acid (EPA) may help to promote body weight loss and reduce inflammatory and cardiovascular disease-related risk markers in conjunction with hypocaloric diets. The present study aimed to assess methylation modifications in white blood cell genes induced by EPA and/or LA supplementation, as well as their possible relationship with metabolic risk biomarkers in obesity during weight loss.

**Methods:** Apparently healthy overweight/obese women followed a hypocaloric diet (-30%) during 10 weeks after being randomly assigned to 1 of 4 parallel dietary groups: a control group or groups supplemented with 1.3 g EPA/day, 0.3 g LA/day, or both. DNA from white blood cells was hybridized in an Infinium Human 450K methylation microarray.

**Results:** Differentially methylated changes (after-before) were identified in EPA (228 CpGs), LA (125 CpGs) and EPA+LA (279 CpGs) groups as compared with control group. These included CpG regions from NCK adaptor protein 2 (NCK2), fat storage inducing transmembrane protein 2 (FITM2), transcription domain-associated protein (TRRAP), regulatory associated protein of mTOR (RPTOR) and CREB binding protein (CREBBP). Of note, LA significantly upregulated NCK2, TRRAP and RPTOR mRNA in PBMC, which correlated with changes in body weight and fat mass. The variations in cg10320884 (TRRAP) methylation site were negatively associated with changes in TRRAP gene expression, while had a highly significant positive correlation with the changes in the Framingham score.

**Conclusions:** These results suggest the involvement of epigenetic mechanisms in LA and EPA metabolic actions.
**Keywords:** Epigenetics, Lipoic acid, Methylation, Omega-3, Weight loss

**Further collaborators:**

This work was supported by the Ministry of Economy and
Competitiveness from the Government of Spain [ref. AGL 2009-
10873/ALI, BFU2012-36089 and AGL2013-45554-R], Navarra
Government [Department of Health ref. 67/2015], and CIBER-
obn. AE Huerta was supported by a predoctoral grant from "Aso-
ciación de Amigos de la Universidad de Navarra".

144/324

EARLY LIFE PROGRAMMING BY OMEGA-3 FAT-
TY ACIDS IN OBESITY-ASSOCIATED INFLAM-
MATION

(1)Ramalingam, Latha; (2)Menikdiwela, Kalhara; (2)Allen, London;

(2)Scoggin, Shane; (2)Moustaid-Moussa, Naima.

(1)PhD, Nutritional sciences, Texas Tech; (2)Nutritional Sciences, Texas Tech.

**Background and objectives:** Over half of American women of child bearing age are either obese or overweight. Hence, the prevalence of childhood obesity is predicted to increase. Maternal programming through diet is critical for prevention of several diseases in the offspring, including childhood obesity. Clinical trials with n-3 polyunsaturated fatty acids (PUFAs) such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) reported various health benefits, including reduced serum triglycerides and obesity-associated inflammation, and increased insulin sensitivity. However, it remains unclear whether maternal consumption of n-3 PUFAs can protect offspring from the adverse metabolic effects of consuming high fat diets during pregnancy. Accordingly, we hypothesized that n-3 PUFAs supplementation during pregnancy will reduce maternal obesity- associated inflammation and reduce/prevent adverse metabolic effects in offspring.

**Methods:** To test this hypothesis, we fed B6 mice high fat diet (HF) or HF supplemented with fish oil, rich with EPA (HF-EPA) for 8 weeks pre-pregnancy through lactation. After weaning, offspring from HF or HF-EPA dams were either continued on their respective diet or switched to the opposite diet. Five treatment groups were created including a low fat (LF)-LF control group (LF maternal diet and LF post-weaning), HF-HF, HF-EPA, EPA-HF, EPA-EPA, respectively.

**Results:** Metabolic phenotyping of these mice included body weight and body fat, glucose and insulin tolerance tests. Offspring born to dams fed EPA and continued on EPA (EPA-EPA) showed significantly higher glucose clearance rates compared to other treatment groups but was lower compared to LF animals. Similarly, significant improvements in insulin sensitivity was observed in the EPA-EPA fed group compared to other HF treated groups. EPA-EPA and HF-EPA had lower fat mass compared to HF-HF and EPA-HF. Additional mechanistic studies are ongoing in serum and tissues collected from the offspring to determine molecular

mechanisms mediating EPA effects on high fat diet-induced obe-
sity and inflammation in dams and their offspring.

**Conclusions:** In conclusion, these studies will help to understand the differential contribution of maternal dietary lipids to metabolic health of the offspring.

**Keywords:** Obesity, Eicosapentaenoic acid (EPA), fish oil, mice, in utero, childhood obesity

144/340

ANGIOTENSIN I-CONVERTING ENZYME INHIB-
ITORY ACTIVITY OF OAT PROTEINS HYDRO-
LYSATES

(1)Darewicz, Malgorzata; (2)Borawska, Justyna; (3)Pliszka, Monika;

(1)Iwaniak, Anna; (4)Minkiewicz, Piotr.

(1)Prof. Warmia and Mazury University in Olsztyn. Faculty of Food Science. Chair of Biochemistry. Poland; (2)Dr. Warmia and Mazury University in Olsztyn. Faculty of Food Science. Chair of Food Biochemistry. Poland; (3)PhD. Warmia and Mazury University in Olsztyn. Faculty of Food Science. Chair of Food Biochemistry. Poland; (4)Prof. Warmia and Mazury University in Olsztyn. Faculty of Food Science. Chair of Food Biochemistry. Poland.

**Background and objectives:** The increase of interest in food ingredients with specific health-related activities have been observed. Biopeptides derived from food proteins are considered as regulators of e. g. cardiovascular system. The best known bioactive peptides with antihypertensive properties are angiotensin I-converting enzyme (ACE) [EC 3.4.15.1] inhibitors. They play a role in lowering high blood pressure and regulating the cardiovascular system activity. Oats proteins may be the source of biopeptides. Nutritional value of proteins from oat seeds is comparable to meat, eggs and milk proteins. The aim of the study was to examine the angiotensin I-converting enzyme inhibitory activity of oat proteins hydrolysates.

**Methods:** The extracts of oat proteins were taken. The study covered the in silico part – with the use of UniProt database, BIO-PEP database and tools available within (http://www.uwm.edu.pl/biochemia/), Fragment Ion Calculator application and Sequence Specific Retention Calculator application. The next step of analysis was hydrolysis with the use of commercial enzyme preparations. Bioactive peptides were identified using reversed-phase liquid chromatography with mass spectrometry detector (RP-HPLC-ESI-MS) based on the expected retention times. The SDS-PAGE method was used to verify the progress of hydrolysis. The ACE inhibitory activity of obtained hydrolysates was assayed according to method by Jimsheena and Gowda (2009).

**Results:** It was found that the degree of hydrolysis and ACE inhibition increased during the hydrolysis process. The sample treated with commercial digestive enzymes demonstrated the highest degree of ACE inhibition (89%, IC50 = 0.42 mg/ml). The degree of ACE inhibition of this sample was higher in comparison with the blank sample (49%, IC50 = 24.89 mg/ml). The proteins
are only partially hydrolysed with pepsin in the stomach, so the
sample hydrolyzed by pepsin was characterized by a lower degree
of inhibition (77%, IC50 = 4.87 mg/ml) in comparison with the
'gastro-duodenal' samples

**Conclusions:** The results showed that the oat proteins can be the source of peptides with ACE inhibitory activity that are released during the simulated digestion. The identification of ACE inhibitory peptides in hydrolysates of oat proteins was possible thanks to the combination of computer and experimental methods.

Keywords: ACE, inhibitory peptides, oat proteins

quences with experimentally confirmed bioactivities as well as
calculation functions helping to evaluate the protein value as the
source of biopeptides.

**Conclusions:** To date, apart from the data concerning different biological properties of peptides, BIOPEP may serve as a tool supporting the experimental and theoretical studies on food-derived biopeptides.

**Keywords:** bioactive peptides; database; food proteins; hydrolysis

144/341

THE BIOPEP - DATABASE OF FOOD BIOLOGICALLY ACTIVE PEPTIDES AND PROTEINS

(1)Darewicz, Malgorzata; (2)Minkiewicz, Piotr; (2)Iwaniak, Anna.

(1)Prof. Warmia and Mazury University in Olsztyn. Faculty of Food Science. Chair of Biochemistry. Poland; (2)Prof. Warmia and Mazury University in Olsztyn. Faculty of Food Science. Chair of Food Biochemistry. Poland.

**Background and objectives:** Peptides derived from food proteins affect biological, functional, immunological and sensory properties of food products as well as provide information concerning product composition, authenticity or history. Proteins apart from their function as the nutrients act as the precursors of peptides with variety of functions.

**Methods:** The database of protein and peptide sequences called BIOPEP (http://www.uwm.edu.pl/biochemia) has been designed mainly for scientists working in the area of food and nutrition. BIOPEP database consists of four sequence databases: proteins, bioactive peptides, allergenic peptides with their epitopes and sensory peptides. The information concerning peptide or protein covers its sequence; data about activity or taste; references or in the case of allergenic protein database reference, sequence of experimental and theoretically predicted linear epitopes [Minkiewicz et al. 2012]. Sequence analysis options include: profiles of the potential biological activity, epitopes or sensory activity, calculation of the quantitative parameters A and B useful for evaluation and classification of proteins as precursors of bioactive or sensory peptides as well as immunogenic fragments. The options available in particular databases of BIOPEP include also the simulation and design of proteolysis as well as data mining. BIOPEP contains also the collection of links to other databases and programs ("Useful links" bookmark).

**Results:** Proposed workflows for use of BIOPEP database of bioactive peptide sequences cover among others: using sequences of peptides as queries for database screening or identification of peptides from BIOPEP among products of protein hydrolysis. Selected examples of applications of the database, described by other authors will be presented. Our intention was to establish BIOPEP as the database integrating information about various properties of fragments originating from food proteins. All subelements of the BIOPEP database are continuously updated with the new sequences

144/401

ANTIHYPOXIA AND ANTIOXIDANT EFFECTS OF SMALL MOLECULE OLIGOPEPTIDES ISOLATED FROM PANAX GINSENG C. A. MEYER

(1)Li, Yong; (2)Ren, Jinwei; (3)Li, Di; (3)Xu, Meihong.

(1)Prof. Nutritionist. Department of Nutrition and Food Hygiene, School of Public Health, Peking University, Beijing 100191;

(2)Msc. Nutritionist. Department of Nutrition and Food Hygiene, School of Public Health, Peking University, Beijing 100191; (3)PhD. Nutritionist. Department of Nutrition and Food Hygiene, School of Public Health, Peking University, Beijing 100191.

**Background and objectives:** To investigate the effect of Panax ginseng oligopeptide of Jilin providence (GOP) on antioxidation in oxidative damaged rats induced by D-galactose and antihypoxia capacity in mice.

**Methods:** 90 specific pathogen free (SPF) male SD rats are randomly divided into 9 groups: namely blank control group, model control group, whey protein control group, and six groups of GOP at different doses. Except the control group, all other groups are given with 125 mg/kg of D-galactose by intraperitoneal injection for 6 weeks. Then, rats in the six samples groups were gavaged with 0.0625, 0.125, 0.250, 0.500, 1.000, 2.000 g/kg of GOP; rats in control group and model group were gavaged with the same volume of distilled water; and rats in whey protein group were gavaged with 0.250 g/kg of whey protein for 45 d. At the same time, rats are given with D-galactose uninterruptedly. The oxidative stress parameters in serum and liver tissue are then determined. Besides, 180 SPF male mice are randomly divided into three experimental sets for the detection of different indicators. Each set of mice are then randomly divided into 6 groups: namely blank control group, whey protein control group, and four groups of GOP at different doses. Mice in the four samples groups were gavaged with 0.075, 0.150, 0.300, 0.600 g/kg of GOP; mice in bland control group was gavaged with the same volume of distilled water; and mice in whey protein control group were gavaged with 0.300 g/kg of whey protein for 30 d. The survival time of oxygen deprivation, intoxication and cerebral ischemic hypoxia of mice are then determined.

**Results:** The levels of lipid peroxide and protein peroxide of GOP groups are significantly lower than those of model group, meanwhile, the activities of antioxidant enzyme and the levels of antioxidants significantly increase compared with these of model group, and the effects are better than those of the whey protein.
The survival time of oxygen deprivation, intoxication and cerebral ischemic hypoxia of mice are significantly longer than these of whey protein control group.

**Conclusions:** GOP has antioxidant effect on D-galactose induced oxidative damaged rats as well as antihypoxia capacity on mice.

**Keywords:** Panax ginseng oligopeptide of Jilin providence; antioxidant; antihypoxia

**Further collaborators:** Some issue about the peptide nutrition.

## 144/403

# THE INFLUENCE AND MECHANISM OF BOVINE COLLAGEN OLIGOPEPTIDE ON WOUND HEALING IN MICE

(1)Li, Yong; (2)Xu, Teng; (3)Li, Di; (3)Xu, Meihong.

(1)Prof. Nutritionist. Department of Nutrition and Food Hygiene, School of Public Health, Peking University, Beijing 100191;

(2)Msc. Nutritionist. Department of Nutrition and Food Hygiene, School of Public Health, Peking University, Beijing 100191; (3)PhD. Nutritionist. Department of Nutrition and Food Hygiene, School of Public Health, Peking University, Beijing 100191.

**Background and objectives:** To investigate the influence and mechanism of bovine collagen oligopeptide on wound healing in mice.

**Methods:** A total of 240 ICR mice are randomly assigned to 6 groups, including a control group and 6 bovine collagen oligopeptide intervention groups (0.375, 0.75, 1.5, 3.0, 6.0g/kg bw). And each group is randomly assigned to 4 subgroups. The mice are carried out by mice stomach-lavaging continuously for 16 days after excisional wound model. And the mice will be sacrificed separately at day 4, day8, day12 and day 16. Wound closure is assayed, and the serum SDF-1α parameter is conducted. After fixation of the skin biopsy samples in 4% formalin, they are embedded in paraffin and stained with HE.

**Results:** It has been found that Bovine collagen oligopeptide treatment could significantly decrease the wound region (p<0.05 or p<0.01) and increase the secret of SDF-1α(p<0.05 or p<0.01).

**Conclusions:** These findings suggest that Bovine collagen oligopeptide have an obvious promote on wound healing and the underlying mechanism may be related to enhancing SDF-1α.

**Keywords:** Bovine collagen oligopeptide; wound healing; SDF-1α

**Further collaborators:** Some issue about the peptide nutrition.

## 144/404

# THE EFFECTS OF WALNUT PEPTIDE ON LEARNING AND MEMORY ABILITIES IN YOUNG MICE

(1)Li, Yong; (2)Du, Qian; (3)Fan, Rui; (3)Xu, Meihong.

(1)Prof. Nutritionist. Department of Nutrition and Food Hygiene, School of Public Health, Peking University, Beijing 100191;

(2)Msc. Nutritionist. Department of Nutrition and Food Hygiene, School of Public Health, Peking University, Beijing 100191; (3)PhD. Nutritionist. Department of Nutrition and Food Hygiene, School of Public Health, Peking University, Beijing 100191.

**Background and objectives:** To investigate the improvement effects and their mechanisms of walnut peptide (WP) on learning and memory abilities in young mice.

**Methods:** 48 male weaning C57BL/6J mice were randomly divided into 4 groups, the control group, low WP intervention group, middle WP intervention group, and high WP intervention group(the concentrations of WP were 110mg/kg.bw, 220mg/kg.bw and 440mg/kg.bw respectively), with 12 mice in each group. After 30 days feeding, the Open-field test, Morris Water Maze, Step-down test and Shuttle test were performed, in order to observe the ability of autonomic activities and emotional responses, spatial memory ability, and the active and passive avoidance reactions in mice.

**Results:** According to the results of Morris Water maze, the escape latency of high WP group was significantly reduced compared with the control group (P<0.05), and for middle and high WP groups, the times staying in the target quadrant and the numbers of crossing over the position where the platform had been were increased compared with the control group (P<0.05). The Step-down test indicated, after the training 24 hours, compared to the control group, the low and high WP groups had less error times (P<0.05), and three intervention groups had longer step-down latency (P<0.05). After the training 5 days, the declines of learning and memory in the middle WP group was effectively reduced (P<0.05). Shuttle test showed that, compared with the control group, after the intervention of the low, middle, and high doses of WP, the numbers of active avoidance reaction increased (P<0.05), while the times of electric shock decreased in mice (P<0.05). The activities of T-SOD and GSH-Px in three WP groups were higher, and the level of MDA were lower compared with the control group (P<0.05).

**Conclusions:** The walnut Peptide could enhance the abilities of learning and spatial memory and the abilities of active and passive avoidance in young mice, and WP could delay the declines of learning and memory in mice. The mechanism was related to the antioxygenic property of WP.

**Keywords:** Walnut Peptide; Young mice; Learning and memory ability

**Further collaborators:**

Some issue about the peptide nutrition.
# EFFECT OF POWDER DRY ADZUKI BEAN PASTE IN PLACE OF WHEAT FLOUR ON RESISTANT STARCH CONTENT AND SENSORY CHARACTERISTICS OF COOKIES

(¹)Kamei, Aya; (²)Ito, Tomomi.

(¹)PhD. Division of Domestic Science Education. Faculty of Education. Miyagi University of Education. Japan.; (²)BEd. Division of Domestic Science Education. Faculty of Education. Miyagi University of Education. Japan.

**Background and objectives:** Traditional Japanese sweets have been popular among Japanese, and are often eaten particularly by middle-aged and elderly people. A lot of traditional Japanese sweets contain adzuki bean paste(ABP). Adzuki beans are a good source of carbohydrate as well as of protein, because these are starchy pulse. Furthermore, they contain a lot of dietary fiber and also include resistant starch (RS). RS escapes digestion until reaching colon and acts like dietary fiber. Recently, many studies recommend taking this new type of dietary fiber for health benefits. The purpose of this study was to investigate feasibility of replacing wheat flour to powder dry ABP by analyzing RS contents and sensory characteristics in cookies.

**Methods:** Cookies were prepared from various blends of wheat flour(WF) and powder dry ABP (100%WF:0%ABP, 50%WF:50%ABP, 25%WF:75%ABP, 0%WF:100%ABP). RS content of these cookies were measured by Resistant Starch Assay Kit (Megazyme International Ireland, Ireland) and ranking test was applied for evaluating sensory characteristics.

**Results:** RS content of powder dry ABP was 5.3% and RS contents of cookies blended flour with 50%ABP, 75%ABP, 100%ABP were 1.6%, 2.1%, 3.0%, respectively. As a ratio of ABP in the cookies increased, a ratio of RS content also increased. Sensory evaluation indicated that 50%ABP cookies was not different from 0%ABP cookies in terms of flavor, sweetness, texture and overall acceptability.

**Conclusions:** These results implied that cookies made of powder dry ABP in place of wheat flour increased amount of RS in cookies. Taking account of sensory evaluation, 50% replacement of powder dry ABP cookies were found to be acceptable.

**Keywords:** dry adzuki bean paste, resistant starch, sensory characteristics, cookies

# THE PHYSIOTHERAPIST AND THE FUNCTIONAL FOODS IN THE QUALITY OF LIFE IN HEALTH

(¹)Mortágua, Andréa Rebouças; (²)Ribeiro, Euler Esteves; (³)Ribeiro, Ednéa Aguiar Maia; (⁴)de Souza Praia, Raquel; (⁵)Oneti, Ciro Félix;

(⁶)Costa, Michelle Silva; (⁷)Araújo, Arthenize Riame Praia G. C.; (⁸)Neta, Inez Siqueira Santiago; (⁹)Mesquita, Priscila Lyra; (¹⁰)Nascimento, Priscilla Hossaine Santos do.

(¹)Fisioterapeuta do Projeto piloto Saúde em Casa. Universidade Aberta da Terceira Idade (UNATI/UEA). Manaus/AM/Brasil.;

(²)Médico. Doutorado em Medicina e Ciências da Saúde. Diretor da Universidade Aberta da Terceira Idade (UNATI/UEA). Manaus/AM/Brasil.; (³)Médica. Doutorado em Ciências da Atividade Física e Desporte. Universidade Aberta da Terceira Idade (Manaus/AM). Brasil.; (⁴)Doutoranda em Saúde Pública. Universidade de Ciências Empresariales y Sociales (UCES). Enfermeira. Universidade Aberta da Terceira Idade (Manaus/AM). Brasil.; (⁵)Enfermeiro. Universidade Aberta da Terceira Idade (Manaus/AM). Brasil.;

(⁶)Assistente Social. Especialista em Política Pública de Atenção a Família. Universidade Aberta da Terceira Idade (Manaus/AM). Brasil.; (⁷)Acadêmica de Enfermagem. Universidade Paulista (UNIP/AM). Universidade Aberta da Terceira Idade (MANAUS/AM). Brasil.; (⁸)Acadêmica de Enfermagem. Universidade Federal do Amazonas. Universidade Aberta da Terceira Idade (MANAUS/AM). Brasil.; (⁹)Educadora Física. Secretaria de Educação do Estado de São Paulo.; (¹⁰)Assistente Social. Especialista em Assistência e Família. Universidade Aberta da Terceira Idade (Manaus/AM). Brasil.

**Background and objectives:** The Unified Health System (SUS), whose principles are the pursuit of universality, integrality and equity, forming several productions and questions regarding the assistance adopted. There is a need for more active professionals in the epidemiological reality inserted in the SUS, in this context the Physiotherapy is inserted in the system with an exclusively rehabilitating and curative character. Objective: To analyze the main actions of the physiotherapist of the multiprofessional team, aimed at promoting healthy eating habits.

**Methods:** This is a literature review in the SciELO (Scientific Electronic Library Online) and LILACS (Latin American and Caribbean Literature in Health Sciences) databases, with the descriptors in health: functional foods AND physiotherapy. Criteria to inclusion and exclusion were applied, and 4 studies were selected for analysis.

**Results:** The increase of obesity has caused great concern to sanitary skills, presenting as a significant health risk factor, such as: diabetes, hypertension, hypercholesterolemia, among others. Physiotherapy in its work with regard to nutrition aims to promote and raise awareness in the community in promoting healthy eating habits and performance of physical exercises. A balanced diet is essential to health by providing the necessary nutrients and functional foods are the newest frontiers in the food market, as they still perform therapeutic functions. In Brazil 75% of the food sold is linked to health, this audience is divided into classes of A & B, present in the market, among them
we can mention: yogurts with pro-biotics that regulate intesti-
nal function, iron-enriched milks that help in prevention and in
treatments Anemias, vitamins and with Omega 3 acid, highlight-
ing eggs and margarines enriched also with Omega-3, among
several other foods.

**Conclusions:** Functional food has low cost and is of great nu-
tritional value, favoring the indication by several professionals in
the health area. In this way, physiotherapy in the search for the
well-being of its patient, has the duty not only to encourage or
accompany the practice of exercises, but also to plan actions relat-
ed to the theme, together with the multiprofessional team, actions
in the prevention and promotion of Health of the individual or
community.

**Keywords:** Physiotherapist, Functional foods, Promotion of health.

144/470

EXPLORATION OF BIOFUNCTION OF SIPHO-
NAXANTHIN, A CAROTENOID DERIVED FROM
GREEN ALGAE, ON HEPATIC LIPOGENESIS

(1)Zheng, Jiawen; (2)Li, Zhuosi; (3)Manabe, Yuki; (2)Kim, Minji; (2)Goto, Tsuyoshi; (2)Kawada, Teruo; (2)Sugawara, Tatsuya.

(1)Kyoto University; (2)Phd. Graduate School of Agriculture. Kyoto University. Japan; (3)MSc. Graduate School of Agriculture. Kyoto University. Japan.

**Background and objectives:** Non-alcoholic fatty liver disease (NAFLD) is notable for its high morbidity and mortality accompanied by a series of complications including diabetes, hypertension, dyslipidemia etc. Carotenoids have been well-known for their various activities, such as anti-oxidation, anti-photoaging, anti-angiogenesis and anti-obesity. In concern with the diverse functions of carotenoids, their potential to prevent and alleviate NAFLD has also been investigated. Siphonaxanthin is a rare marine carotenoid present in some green algae. In this research, we investigated the potential function of siphonaxanthin on preventing and attenuating NAFLD and its mechanisms.

**Methods:** We conducted a preliminary assessment of the ef-
fect of siphonxanthin on hepatic lipid metabolism by conducting
an in vitro study. HepG2 cell line, derived from human liver can-
cer, was used as a model of liver. We used a synthetic agonist of
liver X receptor α (LXRα) to simulate an aggravated lipogenesis
in HepG2 cells and investigated the effect of siphonaxanthin on
hepatic lipid synthesis from several aspects including triacylglyc-
erol accumulation, mRNA levels of related genes, protein level of a
transcription factor named sterol regulatory element binding pro-
tein-1c (SREBP-1c) and LXRα activity.

**Results:** Siphonxanthin significantly suppressed the excess accumulation of triacylglycerol induced by LXRα agonist by down-regulating the expression of SREBP-1c and a set of genes related to de novo lipogenesis. On the other side, fatty acid translocase (CD36) and fatty acid binding protein-1 (FABP1) which regulate fatty acid uptake also showed a significant decrease in

translational levels. Furthermore, we found out that siphonxan-
thin inhibited LXRα activation induced by its agonist, T0901317.

**Conclusions:** In the present study, we discovered that a rare carotenoid named siphonaxanthin, possesses a strong inhibitory effect on hepatic lipogenesis. Based on this study, it is plausible to speculate that siphonaxanthin may be promising in nutraceutical application targeting pathological liver disorders such as NAFLD.

**Keywords:** Siphonaxanthin, Hepatocyte, Lipogenesis, LXRα, SREBP1c.

144/506

ASSOCIATION BETWEEN AMINO ACIDS, BIO-
MARKERS OF PROSTATE CANCER AND INFLAM-
MATION IN NORWEGIAN PROSTATE CANCER
PATIENTS

(1)Rangel Huerta, Oscar Daniel; (2)Paur, Ingvild; (2)Bastani, Nasser;

(2)Kjølsrud Bøhn, Siv; (3)Lilleby, Wolfgang; (4)Smeland, Sigbjørn;

(2)Karlsen, Anette S.; (2)Blomhoff, Rune.

(1)PhD. Department of Nutrition. Institute of Basic Medical Sciences. University of Oslo.; (2)PhD. Department of Nutrition. Institute of Basic Medical Sciences. University of Oslo. Norway.;

(3)MD.Division of Cancer Medicine. Transplantation and Surgery. Oslo University Hospital. Norway.; (4)PhD.Division of Cancer Medicine. Transplantation and Surgery. Oslo University Hospital. Norway.

**Background and objectives:** Earlier our research group had shown that a nutritional intervention with tomato products alone or in combination with selenium and n-3 fatty acids soy isoflavones, grape/pomegranate juice, and green/black tea lower prostate-specific antigen (PSA) in patients with non-metastatic prostate cancer. However, the mechanism is still undefined. Dysregulation of amino acids has been reported in cancers. Therefore, our aim was to study the amino acid profile in prostate cancer patients following a tomato products-based intervention in relation to PSA and inflammation.

**Methods:** For this analysis, we included 74 patients with pro-
tate cancer from a previous study. Prior to curative treatment, the
patients were randomized to nutritional interventions with either
1) tomato products containing 30 mg lycopene per day; 2) toma-
to products plus selenium, omega-3 fatty acids, soy isoflavones,
grape/pomegranate juice, and green/black tea (tomato-plus); or
3) control diet for 3 weeks. Plasma samples from baseline and
after the intervention were analysed. The level of total PSA was
determined on the AutoDELFIA automatic immunoassay system.
Plasma carotenoids and amino acids were detected using HPLC.
An ANOVA adjusted for the baseline was performed to evaluate
differences between groups after the dietary intervention. Corre-
lation between parameters was estimated by computing Pearson's
correlation coefficient using the delta (t1-t0). All statistical analy-
es were performed using SPSS 24.

**Results:** For all patients, we found a significant correlation be-
tween the lycopene change and total PSA (r=-0.247; p=0.034) and
ornithine (r=0.238; p=0.041). Furthermore, the change in ornithine was inversely correlated with total PSA (r=0-0.311, p= 0.007). Finally, the change in total PSA was correlated with C-reactive protein (CRP) (r=0.301; p=0.011).

**Conclusions:** The increase of lycopene was associated with an increase of ornithine. The change in ornithine showed to be inversely correlated with PSA, which in turn is associated with the inflammatory biomarker, CRP. Further analyses are needed to study a possible link between ornithine and CRP.

**Keywords:** Carotenoids, Lycopene, Amino acids, Prostate cancer, Inflammation

**Conflict of Interest Disclosure:** RB has interests in Vitas AS. All other authors disclose no conflict of interest.

in natural products: xanthohumol, bergamottin and α-mangostin, all of which contained an isoprenoid side-chain. It is likely that isoprenoid side-chain contributes to PCSK9 inhibition because the basic skeletons of the active compounds are different (phloroglucinol derivative, coumarin, flavonoid, xanthonoid). Of the compounds tested, lupulone showed the strongest PCSK9 inhibitory activity. RNAi experiments showed that knock down of hepatocyte nuclear factor 1α (HNF1α), a positive regulator of PCSK9, impaired the PCSK9 inhibitory activity of lupulone. Furthermore, lupulone decreased HNF1α protein expression in HepG2, suggesting that lupulone inhibits PCSK9 expression through HNF1α downregulation.

**Conclusions:** We isolated and identified PCSK9 inhibitors from food and natural products and suggest HNF1α is involved in the PCSK9 inhibitory mechanisms of lupulone.

**Keywords:** PCSK9, Natural product, HepG2, RNAi, HNF1α.

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## IDENTIFICATION OF PROPROTEIN CONVERTASE SUBTILISIN/KEXIN TYPE 9 (PCSK9) INHIBITORS FROM NATURAL PRODUCTS

(1)Hirata, Hiroshi; (2)Kontani, Ami; (3)Takazumi, Koji; (4)Segawa, Shuichi; (5)Tsuchiya, Youichi.

(1)PhD. Biochemist. Frontier Laboratories for Value Creation. SAPPORO HOLDINGS LTD.; (2)MSc. Physiologist. Frontier Laboratories for Value Creation. SAPPORO HOLDINGS LTD.;

(3)MSc. Analytical chemist. Frontier Laboratories for Value Creation. SAPPORO HOLDINGS LTD.; (4)PhD. Physiologist. Frontier Laboratories for Value Creation. SAPPORO HOLDINGS LTD.;

(5)PhD. Microbiologist. Frontier Laboratories for Value Creation. SAPPORO HOLDINGS LTD.

**Background and objectives:** Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a new target for lowering serum cholesterol levels. PCSK9 inhibition retards degradation of low-density lipoprotein receptor (LDLR), resulting in enhancement of LDL-cholesterol uptake in the liver. Daily intake of foodstuffs containing compounds which inhibit PCSK9 activity is expected to prevent hypercholesterolemia.

**Methods:** We constructed a food extracts library and screened for PCSK9 inhibition. A human hepatocellular cell line (HepG2) was treated with test samples and cultured for one day. Secreted mature PCSK9 proteins from the culture medium were determined by western blotting. Active compounds in the food extracts responsible for PCSK9 inhibition were separated and isolated by chromatography. Chemical structures of the compounds were determined by LC-Q-TOF-MS and NMR analyses. To investigate the mechanisms of PCSK9 inhibition in the food items, HepG2 cells were transfected with the siRNA of PCSK9 regulators and then treated with the active compound.

**Results:** Female hop cones (Humulus lupulus L.) and lemon peel showed PCSK9 inhibitory activity. The active compounds responsible for inhibition of PCSK9 were identified as lupulone and 5-geranyloxy-7-methoxycoumarin from hop and lemon, respectively. The active compounds contained an isoprenoid side-chain and prenyl and geranyl moieties. We also found PCSK9 inhibitors

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## INVESTIGATION OF MICROBIAL FLORA AND REGIONAL DIFFERENCE OF KUMISS USING ARISA

(1)Takahashi, Soichiro; (2)Yamaguchi, Akihiro; (2)Ishii, Satomi.

(1)Rakuno Gakuen University Graduate Student; (2)Rakuno Gakuen University PhD.

**Background and objectives:** Kumiss is a prepared by fermenting fresh horse milk with some lactic acid bacteria and yeasts in dry inland Asia. Kumiss is carbonated beverage containing about 1 to 2.5 % alcohol traditional ethnic drink for nomads in maintain health. In this study, we investigated regional differences in the type and amount of microorganisms using automated rRNA intergenic spacer analysis (ARISA).

**Methods:** Nucleic acids were extracted from 32 samples of horse kumiss which was transported in frozen state from Mongolia using an automated nucleic acid extractor (magLEAD). The extracted DNA was amplified by PCR using Bac-2 primer set and Fun-1 primer set. Agarose gel electrophoresis was carried out to confirm the PCR amplification band. After that, we requested fragment analysis of PCR product and estimated the kind and quantity of microbial species from the obtained data. We investigated the relationship between these and regional differences.

**Results:** Similar results were obtained in kumiss in the same area. The presence of various bacteria and fungi was revealed in any kumiss. In many samples there were intense signals of bacteria and fungi near ARISA fragment length (bq) 305 and 701, respectively, whereas samples showing different peaks existed. They were in the range of ± 0.1%, and it was judged that they are the same type of microorganisms.

**Conclusions:** It is suggested that microorganisms different from each other are involved in kumiss. In the future, we will conduct identification tests of microorganisms in which signal has appeared strongly and proceed with identification of bacterial species. We will also investigate the relationship with general ingredients and analyze the metabolic products of bacteria contained in kumiss.

**Keywords:** Kumiss, Arisa, Inland Asia, Bacteria.
# EVALUATION OF KNOWLEDGE AND CONSUMPTION OF FUNCTIONAL FOODS BY ADOLESCENTS

(1)Aranha, Flávia; (2)Barretto, Daniela; (2)Burriguel, Gabriela;

(2)Mendes, Juliana; (2)Guidorizi, Luana; (2)Barchi, Natalia.

(1)PhD. Department of Education. Institute of Biosciences. UNESP. Univ Estadual Paulista, Câmpus Botucatu. Brazil.; (2)BSc. Graduation Course in Nutrition. Institute of Biosciences. UNESP. Univ Estadual Paulista. Brazil.

**Background and objectives:** Functional food in addition to its nutritional value inherent in its chemical composition may play a potentially beneficial role in the prevention and treatment of diseases if consumed frequently and in adequate amounts. The objective of this study was to evaluate the knowledge and consumption of functional foods in the school meal service.

**Methods:** A Food Frequency Questionnaire (FFQ) was applied with a focus on functional foods for students aged 15 to 18 years of Technical School (n0 139). The questionnaire was divided into two parts, the first part about knowledge, asked the interviewee if he knew what was a functional food (if the answer was "YES" was asked to explain the definition), and the second part was (Soups, fish, milk / dairy products, vegetables / fruits, eggs, sauces / seasonings, and soy / dairy products) were used on a daily basis (daily, weekly, monthly or never). After completing the questionnaire, a lecture was given explaining and exemplifying the definition and importance of the consumption of these foods.

**Results:** Regarding the results of the first part of the questionnaire: Do you know what functional food is? The adolescents answered 93.5% No and 6.47% Yes. To whom answered Yes was asked the second question: Do you know the importance of functional foods for your health? The students answered: "Foods that help in the proper functioning of the body or organism". The second part of the questionnaire was about the consumption of functional foods (divided into groups and their frequency), we can observe the following results: Soups (70%) and fish (81%) consumption is monthly; Dairy products (69%), vegetables (42%), fruits, eggs (77%), sauces (72%) and seasonings (73%) weekly consumption and soybeans / derivatives (70%) never consume. In relation to the lecture given the adolescents were very interested in the discussion, commenting and asking a lot about the food and its functional action in the body.

**Conclusions:** The interviewees' lack of knowledge about functional foods was visible, but the interviewed population frequently consumes some foods with functional properties.

**Keywords:** Feeding, Health, Composite bioactive, Diners.

# THE EFFECTS OF RESISTANT STARCH ON BONE LOSS VIA INTESTINAL MICROBIOTA AND INFLAMMATION IN OVARIECTOMISED MICE

(1)Tousen, Yuko; (2)Matsumoto, Yu; (3)Nagahata, Yuya; (2)Yoshiko, Ishimi.

(1)PhD. Researcher. Department of Food Function and Labeling. National institute of Health and Nutrition. National Institutes of Biomedical Innovation. Health and Nutrition. Tokyo.; (2)PhD. Researcher. Department of Food Function and Labeling. National institute of Health and Nutrition. National Institutes of Biomedical Innovation, Health and Nutrition. Japan.; (3)MSc. Researcher. Product Development Laboratory. J-OIL MILS. Inc. Japan.

**Background and objectives:** Recent studies have suggested an important role for gut-bone signalling pathways and microbiota in regulating bone health through modification of the immune status. Altered immune status associated with oestrogen deficiency can lead to bone loss; however, the intestinal microbiota can modulate the host metabolism and immune status. Resistant starch (RS) is a type of dietary fibre that is not absorbed in the small intestine of healthy humans, and is fermented to a large extent by microbiota in the colon, resulting in changes in the composition of gut microbiota and pre-biotic effects. Therefore, we hypothesised that intake of RS modulates intestinal microbiota and inflammation of the intestine and bone, resulting in reduced bone loss caused by oestrogen deficiency. The present study evaluated the effect of diet supplemented with RS on intestinal microbiota, bone mineral density (BMD), and inflammatory gene expression in the colon and bone marrow of ovariectomised mice.

**Methods:** Female ddY strain mice, aged 8 weeks, were either sham-operated (Sham, n = 8) or OVX. OVX mice were randomly divided into the following three groups (8 per group): OVX control (OVX); OVX fed 20 % high amylose cornstarch (HAS) diet (OVX + HAS); OVX fed 20 % acid-hydrolysed high amylose cornstarch (AHAS) diet. AHAS was more resistant to enzymatic digestion than HAS. RS was included as 34% or 60% of dry weight in HAS or AHAS, respectively.

**Results:** After 6 weeks, treatment with HAS and/or AHAS accelerated fermentation and increased the β-glucosidase activity in the cecum, and increased the rate of Bifidobacterium spp. in the faeces. The mRNA of the anti-inflammatory cytokine IL-10 tended to be up-regulated in the large intestine by treatment with AHAS. Conversely, the mRNAs of the osteoclastogenic cytokine RANKL and inflammation-related IL-7R were downregulated in the bone marrow by treatment with AHAS. Treatment with AHAS protected against OVX-induced bone loss.

**Conclusions:** Our findings suggest that an AHAS diet might alter the microbiota and immune status of the large intestine and bone marrow, resulting in attenuated bone resorption in OVX mice.

**Keywords:** Osteoporosis. Resistant starch. Microbiota. Inflammation. Bone.
# EFFECT OF POMEGRANATE JUICE CONSUMPTION ON THE LEVELS OF GLUCOSE AND LIPIDIC PROFILE

(1)Valdés Miramontes, Elia Herminia; (2)Virgen-Carrillo, Carmen Alejandrina; (3)Martínez-Moreno, Alma Gabriela; (4)Castañeda-Saucedo, María Claudia.

(1)PhD. Chemistry pharmacobiology. Centro de Investigaciones en Comportamiento Alimentario y Nutrición. CUSur. University of Guadalajara. Mexico.; (2)BSc. Physician. Centro de Investigaciones en Comportamiento Alimentario y Nutrición. CUSur. University of Guadalajara. Mexico.; (3)Ph D. Psychologist. Centro de Investigaciones en Comportamiento Alimentario y Nutrición. CUSur. University of Guadalajara. Mexico.; (4)Ph D. Agronomist. Centro de Investigación en Estudios Territoriales y Ruralidad. University of Guadalajara. Mexico.

**Background and objectives:** Diabetes mellitus represents the second cause of mortality in Mexico and its prevalence continues to increase. Nowadays, an increased interest of functional foods as an alternative treatment is taken place. It has been proposed that pomegranate, through its high content of phenolic compounds, is capable of reducing glucose and also possesses antiatherogenic effects. The aim of this research was to evaluate the consumption of pomegranate juice and its effect on glycemia and lipid profile in diabetic rats.

**Methods:** A longitudinal and experimental study was performed with 18 wistar rats induced hyperglycemia by streptozotocin and divided into 3 groups exposed to standard diet and water. In addition, the following interventions: group 1 (G1) pomegranate juice. Groups 2 (G2) and 3 (G3) solution with sucrose at the same concentration as the brix degrees of juice. G3 was also given insulin treatment. Liquid consumption was recorded every 24 hours for 28 days. Plasma levels of glucose, cholesterol and triglycerides were measured every 7 days.

**Results:** An analysis of variance (ANOVA) was performed with a Tukey mean comparison test. G1 consumed on average 33.59 ml of juice. Groups G2 and G3 consumed on average 77.6 and 82.8 ml of sucrose solution respectively. The average glycemia levels at the beginning and the end of the intervention were for G1: 417-365mg/dl, G2: 461-383mg/dl, G3: 429-214mg/dl respectively. At the end, for glucose concentration, the ANOVA showed a statistically significant difference between the groups (p <0.05). The average cholesterol was for G1: 79mg/dl, G2: 92mg/dl, G3: 78mg/dl. The mean triglycerides for G1: 180mg/dl, G2: 248mg/dl, G3: 169mg/dl. No statistically significant differences were found between cholesterol and triglycerides in the groups.

**Conclusions:** Pomegranate juice had a lower degree of preference with respect to a sucrose solution. In spite of this, the group exposed to juice had lower blood glucose levels, and statistically significant difference between groups were obtained. This suggests that pomegranate juice may act as an alternative complement during the treatment of hyperglycemia.

**Keywords:** Diabetes mellitus, Pomegranate juice, Consumption, Glycemia, Lipid profile.

# EFFECT OF MEDICINAL HERB HANGRYON ON GHRELIN SECRETION OF GASTROENTERIC-BRAIN HORMONE MECHNISM

(1)Kim, Inho; (2)Han, Taewon; (1)Lee, Changho; (1)Han, Daeseok.

(1)Ph. D. Functionality Evaluation Research Group of Korea Food Research Institute. Republic of Korea.; (2)MSc. Functionality Evaluation Research Group of Korea Food Research Institute. Republic of Korea.

**Background and objectives:** Korean medicinal herb Hangryon extract has applied to ghrelin function on gastroenteric-brain hormone mechanism. The ghrelin is a peptide hormone produced by stomach cells in gastrointestinal tract circulating to hypothalamic area through blood brain barrier which regulates food intake.

**Methods:** Tests have been conducted to use human microvascular endothelial cells (HMVECs) and male Sprague-Dawley (SD) rats in in vitro and in vivo models respectively. Ghrelin was measured by enzyme immunoassay (EIA) in cell culture supernatant or rat serum.

**Results:** The ghrelin concentration was significantly decreased to 92 ± 2%, in vitro and 73 ± 9%, in vivo compared to control group when Hangryon extracts was applied to model systems. It means low ghrelin level affects to satiety signal and appetite control. Physicochemical characteristics of ghrelin level with sample extract on the extract were tested on cell viability and hormone level in various pH and temperature conditions. Concentration of ghrelin hormone has shown 91% to 96% on pH change 2 to 10 and 92% to 95% on 60°C to 100°C. Cell proliferation was 94% to 100% on sample concentration at 500 µg/mL.

**Conclusions:** It results that the sample has no cytotoxicity as well as inhibition rate of sample extract was no difference significantly and stable on ghrelin suppression in pH or temperature changes. That also suggested that natural medicinal herb Hangryon would be able to developed to functional food ingredient for anti-obesity product related to gastroenteric-brain hormone mechanism with activity maintenance during processing or digestion in our body.

**Keywords:** Medicinal herb, Ghrelin concentration, Cell and animal test, Physicochemical characteristics.

**Conflict of Interest Disclosure:** Abstract title: effect of medicinal herb Hangryon on ghrelin secretion of gastroenteric-brain Hormone mechanism

The authors in above the title declare NO conflict of interest. Corresponding author, In-Ho Kim, Ph.D.
## FOLATE CONTENT IN RED RASPBERRIES

Czarnowska-Kujawska, Marta; Gujska, Elzbieta; Michalak, Joanna.

PhD. Department of Food Science. University of Warmia and Mazury in Olsztyn. Poland.

**Background and objectives:** Raspberries are widely consumed fruits due to their unique flavor and beneficial health properties. For years were used for the treatment of various ailments including diabetes, inflammatory diseases and as antimicrobial agents. Several studies have shown preventive effect of raspberries in a variety of diseases, including cancer, diabetes, obesity, cardiovascular diseases, macular degeneration, neurodegeneration as well as inflammation and oxidative stress (Carvalho et al., 2013). Most of the research on raspberries focus on their antioxidant properties together with polyphenolic compounds, vitamin C and tocopherols. However, it is of special interest to analyze another health beneficial compounds contents. Therefore, the aim of the study was to determine folate, vitamin B, content in fresh red raspberries.

**Methods:** Test material (10 samples of raspberries from different cultivators) was obtained from the local fruit processing plant during the harvest period. Folate content was determined according to Czarnowska & Gujska (2012) using HPLC method.

**Results:** Based on the chromatographic separation of folate standards, two folate derivatives, 5-methyltetrahydrofolate and tetrahydrofolate were identified in raspberries samples. The total folate content expressed as folic acid, ranged from 37.6 to 69.5 µg/ 100 g fresh weight. According to the European Food Safety Authority (EFSA), a Population Reference Intake for folate was established at the level of 330 µg/ day, with an increase to 600 µg/ day during pregnancy or lactation (EFSA, 2014).

**Conclusions:** Efforts are being directed to improve the nutritional value of raspberries by breeding and biotechnological programs. Therefore, it is important to provide information on the individual bioactive components, including vitamins, which can be found in these fruits. Our study has shown, that the consumption of fresh raspberries can contribute well to the daily intake of folate, which deficiency is common worldwide.

Carvalho E., Fraser P.D., Martens S. 2013. Food Chemistry 139, 744-752.

Czarnowska, M., & Gujska, E. (2012). Plant Foods for Human Nutrition, 67, 401-406.

**Keywords:** Raspberries, Folate, HPLC

## A SOLUTION TO NUTRITIONAL PROBLEMS OF CHILDREN AT PUBERTATE PERIOD

(1)Hojageldiyev, Taganmyrat; (2)Arashova, Bahar.

(1)Prof. Pharmacology Department. State Medical University of Turkmenistan. Turkmenistan; (2)Assistant. Pharmacology Department. State Medical University of Turkmenistan. Ashgabat. Turkmenistan.

**Background and objectives:** In recent years the link between diet and health of people has become well established, especially importance of nutrition of children during pubertate period which was one of the main issues of Second International Conference on Nutrition (ICN2) in 2014. This generates the need to develop healthy foods, specifically targeting children which has benefits on children's health as well as their potential in reducing risk of disease. Developing functional foods, instead of mostly consumed unhealthy diets such as fast foods, made by local products will be best choice for school children which will improve physiological functions and health of children.

**Methods:** Chicken meat, sesame oil, "Extracti Prosopis Farcta fluidum", "Extracti Artemisia Absinthi" and 200 µm powder of Pepper chillis that is grown in Bagyr village of Ashgabat city in 2015 crumbled by special Chilli grinding Machine was used in making sausage was used. Pharmaco-toxicological and antioxidant properties of sausage and its additives studied on experimental animals (rabbits, rats, mice). Antioxidant property studied by measuring the concentration of malondialdehyde in the tissues, organs of experimental animals.

**Results:** Experiment results conducted on experimental animals showed that no toxicological and high antioxidant effect of sausage and its additives. Because of the insufficiency of unsaturated fatty acids, which are very important for metabolism, in meat products we used sesame oil to compensate this. Also we used "Extracti Prosopis Farcta Fluidum", "Extracti Artemisia Absinthi" and chili pepper as exchange product for usual xenobiotic products that is used in sausage production and at the same time it will increase antibacterial activity and stability of food. Food that is made by using local products and without synthetic substances will affect to the health of people positively, especially children at pubertate period.

**Conclusions:** Method of preparation of real functional food from chicken meat and local medicinal plants was developed.

**Keywords:** Functional food, Children, Health, Medicinal plants.
## GLABRIDIN INHIBITS MUSCLE ATROPHY THROUGH P38/FOXO3A AND GLUCOCORTICOID RECEPTOR SIGNALS

(1)Ashida, Hitoshi; (2)Kubota, Yusuke; (3)Yoshioka, Yasukiyo.

(1)PhD. Professor. Graduate School of Agricultural Science.
Kobe University. Japan.; (2)Ms. Scientist. Graduate School of
Agricultural Science. Kobe University. Japan.; (3)PhD. Postdoctoral
fellow. Graduate School of Science Technology and Innovation.
Kobe University. Japan.

**Background and objectives:** Deterioration of muscle weight due to muscle atrophy decreased quality of life as one of the social problems. Since skeletal muscle weight is maintained by a balance of the synthesis and degradation of proteins, muscle atrophy caused by this collapses. In this study, we search for the food ingredient which is effective in promoting protein synthesis or preventing protein degradation to improve its balance, and try to elucidate this mechanism.

**Methods:** C2C12 skeletal muscle cells were treated with each food components and 1.0 µCi L-[3,5-3H]-tyrosine in differentiation medium. Protein synthesis was determined to measure the incorporation of radio activity into cellular proteins. Protein degradation was determined to measure the release of radio activity induced by dexamethasone (Dex) after pulse label L-[3,5-3H]-tyrosine. Protein expression and phosphorylation levels were detected by western blotting.

**Results:** Among 15 kinds of food compounds glabridin, 4-hydroxyderricin, and xanthoangelol suppressed the Dex-induced protein degradation. On the other hand, all food compounds tested did not promote the protein synthesis. We focused on glabridin and tried to elucidate underlying molecular mechanism. Glabridin inhibited proteolysis through a decrease in the expression levels of MuRF-1 and Cbl-b, which are ubiquitin ligases involving in muscle atrophy. Glabridin also inhibited Dex-induced phosphorylation levels of Foxo3a in nuclear fraction. It is suggested that Foxo3a translocated in nuclear by phosphorylation, resulting in promoted the expression of ubiquitin ligases, MuRF-1 and Cbl-b. We confirmed that glabridin-rich food materials also inhibit muscle atrophy in mice with the same molecular mechanisms.

**Conclusions:** We found glabridin, 4-hydroxyderricin, and xanthoangelol as the food compounds to improve muscle dysfunction. Of these, glabridin prevented phosphorylation of Foxo3a in nuclear fraction, this would be led to down-expression of ubiquitin ligases MuRF-1 and Cbl-b to prevent Dex-induced protein degradation.

**Keywords:** Glabridin, Muscle atrophy, Glucocorticoid receptor; p38; FOXO3a.

**Further collaborators:**

Dr. Yoko Yamashita

## THERAPEUTIC EFFECTS OF EDIBLE BROWN ALGA UNDARIA PINNATIFIDA (WAKAME) IN THE MICE INFECTED WITH HERPES SIMPLEX VIRUS

(1)Lee, Jung-Bum; (2)Hayashi, Kyoko; (3)Yoshinaga, Keiko; (3)Nakano, Takahisa; (2)Uemura, Kazuhide; (2)Kawahara, Toshio; (2)Hayashi, Toshimitsu.

(1)PhD. Assistant Professor. Graduate School of Medicine and Pharmaceutical Sciences for Research. University of Toyama.
Japan.; (2)PhD. Research Institute of Life and Health Sciences.
Chubu University. Japan.; (3)PhD. Riken Vitamin Co. Ltd. Japan.

**Background and objectives:** Herpes simplex virus type 2 (HSV-2) causes genital herpes that is a lifelong infection in populations worldwide. The development of therapeutic agents for preventing genital herpes has become very important in recent years. In the present study, the in vivo anti-HSV-2 effects of wakame proteins (WT), wakame peptides (WP) prepared by protease treatment of wakame and the major dipeptides of WP were assessed.

**Methods:** Female BALB/c mice were inoculated vaginally with HSV-2. WT, WP and three dipeptides, Val-Tyr (VY), Ile-Tyr (IY) and Phe-Tyr (FY), were administered orally from 3 days prior to virus inoculation until 7 days after inoculation. The vaginal washes collected at 3 days after virus inoculation were subjected to virus titration. Clinical signs of infection and survivors were recorded for 2 weeks. In the other experiments, the anti-HSV-2 activities of sera obtained from the mice administered with WP were evaluated.

**Results:** WT suppressed the lesion scores and enhanced the survival rates of mice as compared with distilled water-administered control mice. WP showed significant effects in the suppression of herpetic lesions, virus yields and death of mice. WP contains VY, IY and FY as the major dipeptide components. Among them, IY and FY exerted higher therapeutic effects in the suppression of herpetic lesion, virus production and death of mice. When the mice were treated with single oral administration of WP, the sera obtained during 0 – 8 hours after administration showed potent anti-HSV-2 activities.

**Conclusions:** When the wakame peptide was orally ingested in HSV-2-infected mice, the peptide exhibited potent therapeutic effects. The dipeptides contained in the wakame peptide might play a pivotal role in those effects.

**Keywords:** Brown alga, Undaria pinnatifida, Herpes simplex virus, Genital infection, Therapeutic effects.
Study 2: Nineteen moderately hypercholesterolemic male adults (TC > 5.2 mmol/L) participated, consuming one soft boiled egg per day for 4 weeks in addition to their habitual diet. Before and at weeks 2 and 4, blood samples were obtained after an overnight fast.

**Results:** Study 1: At the end of intervention, serum concentrations of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) were unchanged, despite the significant increase in the intake of dietary cholesterol. In contrast, a significant increase in high-density lipoprotein cholesterol (HDL-C) and a reduction of the LDL-C/HDL-C ratio were observed. Interestingly, the malondialdehyde modified-LDL (MDA-LDL)/LDL-C ratio was decreased and the LDL oxidation lag time, reflecting the resistance of free-radical-induced LDL lipid peroxidation (ex vivo), was significantly prolonged.

Study 2: At weeks 2 and 4, the subjects' serum concentrations of TC and LDL-C did not increase. The serum MDA-LDL concentrations was significantly decreased and the LDL oxidation lag times (ex vivo) was prolonged after 2 and 4 weeks. At weeks 2 and 4, the subjects' serum lutein + zeaxanthin concentrations were significantly higher than their baseline values and showed both an inverse relation with MDA-LDL and a positive relationship with the LDL oxidation lag time.

**Conclusions:** These studies showed that the additional consumption of one egg per day for 4 weeks did not have adverse effects on serum TC or LDL-C, and it might reduce the susceptibility of LDL to oxidation through an increase in the serum lutein and zeaxanthin concentrations.

**Keywords:** Egg, Dietary cholesterol, Serum lipids, LDL oxidation.

**Conflict of Interest Disclosure:** Y.K., C.T. and E.S. are affiliated with an endowed research department from Kewpie Corporation.

Components such as menthol (mint) or 1,8-cineole (found in laurel, basil, rosemary). Previous studies have reported that intragastric administration of menthol and 1,8-cineole induces thermogenesis in anesthetized mice, however, the detailed mechanism is unclear. TRPM8 mRNA is detected in various tissues, including the gastrointestinal mucosa, and in the vagal afferent nerve. Based on these facts, in this study, we aimed to investigate whether TRPM8 expression in the vagal afferent nerve is involved in the enhancement of thermogenesis induced by intake of TRPM8 agonist.

**Methods:** Male mice (C57BL/6, 7wks) were anesthetized with urethane (1.4 g/kg) and thermistors were set on the tail skin, colon, and intrascapular brown adipose tissue (IBAT). Each temperature was used as an index of heat diffusion, core temperature, and local thermogenesis, respectively. A cannula was then inserted into the stomach or jugular vein to administer the solution. The temperature was measured for 10 min before administrating the 1,8-cineole or vehicle and for 180 min after administration.

**Results:** Intragastric administration of 1,8-cineole increased IBAT and colonic temperatures, and M8-B-treatment (TRPM8 antagonist) inhibited these responses. Intravenous administration of 1,8-cineole also showed similar effects. In vagotomized mice, the responses induced by intragastric administration of 1,8-cineole were attenuated.

**Conclusions:** Vagus nerve are involved in the enhancement of thermogenesis induced by intake of TRPM8 agonists.

**Keywords:** Thermogenesis, TRPM8, Brown adipose tissue, Mice, Vagus nerve.

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INVOLVEMENT OF THE VAGUS NERVE IN THE ENHANCEMENT OF THERMOGENESIS BY INTRAGASTRIC ADMINISTRATION OF TRPM8 AGONIST

(¹)Mori, Noriyuki; (²)Urata, Tomomi; (¹)Fukuwatari, Tsutomu.

(¹)PhD. Department of Nutrition. the University of Shiga Prefecture; (²)MSc. Department of Nutrition. the University of Shiga Prefecture.

**Background and objectives:** Obesity increases risk for various diseases and reducing exceeded body fat stores is important for maintaining good health. Thermogenesis contributes to the maintenance of body temperature, and it requires metabolic fuel availability and contributes to energy balance. Therefore, the enhancement of thermogenesis would be important to reduce body fat accumulation. Transient Receptor Potential Melastatin 8 (TRPM8) is a cold receptor activated by mild cold temperature (< 28°C) and it is involved in cold sensation and thermoregulation. Interestingly, TRPM8 is also activated by edible herb com-

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ENRICHMENT OF PREMIX OF FLOUR SUITABLE FOR CELIAC AND ACCEPTABILITY OF BAKED PRODUCTS

(¹)Morínigo Martínez, Macarena; (²)Cubilla Giménez, Pamela Violeta;

(²)Monsalvo Benítez, María Belén; (²)Acosta Escobar, José.

(¹)Nutritionist. Department of Nutrition Research. Faculty of Chemical Sciences. National University of Asuncion. Paraguay;

(²)Nutritionist. Faculty of Chemical Sciences. National University of Asuncion. Paraguay.

**Background and objectives:** In Paraguay, Law 20.830/98 regulates the enrichment of wheat flour as a strategy for the prevention of anemia. This law does not apply to the premix of flour suitable for celiacs. Anemia in celiac disease is 50%. When enriching foods, one of the criteria states that the organoleptic characteristics must be acceptable or should not be modified. Therefore, our objective was to enrich the premix of flour suitable for celiacs with iron and B vitamins, to evaluate the acceptability of two types of high-demand baked goods (bread and sticks) and to further evaluate compliance with the provisions of law.

**Methods:** In specialized kitchens, we elaborate 2 types of products: bread and sticks, each in two versions, with vitamin mix and without vitamin mix. Acceptability was evaluated using a hedonic scale of 9 points and evaluated the characteristics of color,
odor, taste, texture, hardness, appearance and overall score (sum of the average characteristics/6) of the 4 samples presented. The products were presented in individual packages, labeled in numerical form. The volunteers tested each of the 4 products and subsequently completed a self-administered questionnaire. Student's t-test was used.

**Results:** In 105 volunteers with celiac disease (81% female) the mean age was 34.5 ± 10.9 years, 60% reported having had anemia at some point. The evaluated products had the following mean glob al score: unmixed bread 7.42 ± 1.33 (95%CI 7.17-7.68), bread with mixture 8.00 ± 1.06 (95%CI 7.80-8.21), unmixed stick 7.91 ± 1.05 (95%CI 7.71-8.11), and mixed stick 7.86 ± 1.17 (95%CI 7.64-8.09). The differences were minimal, however, mixed bread was signifi cantly more acceptable than unmixed bread (0.58 points, p=0.001) and between the scores of the sticks there were no significant dif ferences (0.05 points p=0.749). We evaluated the compliance with Law 20.830 / 98 and found that the premix for celiacs complies with the contribution of micronutrients (iron, B1, B2, B3 and folic acid).

**Conclusions:** We found that the bread with mixture had a high acceptability and that when the premixture for celiacs was soaked, it could be complied with what was established in the law

**Keywords:** Enrichment, Flour, Celiac, Baked goods, Accepta-
bility.

**Further collaborators:** Laura Elizabeth González Cespedes

**Results:** We previously indicated that the intestinal absorption of dietary plant glucosylceramide is extremely low (J. Lipid Res. 51, 1761-1769, 2010). In the case of sea cucumber cerebroside, the recovery into lymph was similar to plant glucosylceramide. On the other hand, the lymphatic recovery of sphingoid bases from dietary squid CAEP was about ten times higher than those from glucosylceramide, similar to milk sphingomyelin (Lipids 50, 987, 2015). We previously found that dietary plant glucosylceramide and animal sphingomyelin can accelerate the recoveries of dam aged skin barrier functions (Exp. Dermatol. 21, 448-452, 2012). Dietary supplementation with marine sphingolipids involved in both sea cucumber cerebroside and squid CAEP to hairless mouse also improved the skin barrier function.

**Conclusions:** Our findings contribute to our understanding of the absorption of dietary sphingolipids from marine sources and their effects on the host as a dietary integrant. This work was partly supported by the Program for Promotion of Basic and Applied Researches for Innovations in Biooriented Industry (BRAIN) to TS and the Japan Society for the Promotion of Science (JSPS) KAKENHI (Grant No. 16H04923 to TS, YM and TT; Grant No. 15J01143 to NT).

**Keywords:** Intestinal absorption, Seafoods, Skin barrier, Sphingolipids.

144/985

DIETARY MARINE SPHINGOLIPIDS AS FUNC-
TIONAL FOOD COMPONENTS

(1) Sugawara, Tatsuya; (2) Tomonaga, Nami; (3) Duan, Jingjin;

(4) Manabe, Yuki; (5) Tsuduki, Tsuyoshi; (6) Hirata, Takashi.

(1)PhD. Garaduate School of Agriculture. Kyoto University.
Japan; (2)PhD. Graduate School of Agriculture. Kyoto University.
Japan; (3)PhD. Department of Cardiology. Boston Children's
Hospital. United States.; (4)MSc. Graduate School of Agriculture.
Kyoto University. Japan; (5)PhD. Graduate School of Agricultural
Science. Tohoku University. Japan.; (6)PhD. Shijonawate Gakuen
University. Japan.

**Background and objectives:** Sphingolipids are ubiquitous in all eukaryotic organisms and have attracted attention as physiologically functional lipids. Recently, various physiological functions of dietary sphingolipids, especially improving the skin barrier function, have been demonstrated. Sphingolipids of marine invertebrates have atypical types of chain length, unsaturation, and hydroxylation in sphingoid bases. In addition, sphingolipids having C-P bonds, such as ceramide 2-aminoethylphosphonate (CAEP) are found in marine invertebrates. In this study, we evaluated the absorption and functions of dietary marine sphingolipids to utilize marine sources for functional foods.

**Methods:** The intestinal absorption was evaluated via a lipid absorption assay of lymph from the thoracic duct in SD rats. To investigate the effects of dietary sphingolipids on skin, we characterized the recovery of skin barrier using a special Mg-deficient diet induced atopic dermatitis-like hairless mice model.

144/987

CLINICAL EFFECTS OF EVIDENCE-BASED BIL-
BERRY EXTRACT ON ASTHENOPIA

(1)Kosehira, Marie; (2)Kitaichi, Nobuyoshi.

(1)Chief Researcher. Research and Development Dept. Omnica Co., Ltd. Japan; (2)MD. PhD. Professor and Chairman. Department of Ophthalmology Health Sciences University of Hokkaido. Japan.

**Background and objectives:** We evaluated the effect of evidence-based bilberry extract (EBBE) intake for asthenopia.

**Methods:** Placebo-controlled double-blind randomized clini cal trial was conducted in 30 Japanese healthy volunteers (adults: 20-59 years old). Subjects took 160mg of EBBE (contain 37% Vaccinium myrtillus anthocyanin) or placebo daily for 4 weeks. On the first and the last day of the administration, objective exami nations were performed three times: before, prompt after, and 20 minutes after video display terminal (VDT) work. They played Tetris game for 30 minutes on iPhone as deemed VDT work. Ac commodation was examined by using TriIRIS® C9000.

**Results:** Two of accommodative functions “Miosis” and “Con-vergence” were statistically improved in EBBE group compared with controls at 20 minutes after VDT work (P<0.05 and 0.1).

**Conclusions:** Oral intake of qualified EBBE saved asthenopia objectively when abusing eyes.

**Keywords:** Asthenopia, Miosis, Convergence, Evidence-based bilberry extract, VMA

**Conflict of Interest Disclosure:** Marie Kosehira is an em-
ployee of Omnica Co., Ltd. The other authors have no potential
conflict of interest.
# ORANGE BAGASSE AS AN INGREDIENT IN THE FORMULATION OF A GLUTEN FREE MUFFIN

(¹)Franco Obaldía, Indira del Carmen; (²)Terrero, Paola.

(¹)PhD. Professor. Science and Technology College. Technological University of Panama. Panama.; (²)BSc. Science and Technology College. Technological University of Panama. Panama.

**Background and objectives:** Recently, the number of companies dedicated to the production of natural juices and fruit concentrates is increasing. As a result, an increase in the amount of citrus waste, such as orange bagasse is produced. This orange bagasse can be utilized in animal feed mixes, however it has valuable components for human health, which makes it an appropriate ingredient in the formulation of new food products. The objective of this research is to obtain a nutritional product, gluten free muffin, for a population segment with special needs, as celiac, using as main ingredient the orange bagasse.

**Methods:** Orange bagasse was obtained from a local juice food plant, and it was dried in a solar dryer. Then it was ground, until it was less than 250 microns in size, obtaining dry orange bagasse powder. The orange bagasse was characterized, doing physicochemical and microbiological analyses. Different formulations to obtain muffins were tested, being the most successful, the one using a mix one part corn, one part rice and one part orange bagasse. The muffin was ginger flavoured. Sensorial analyses were developed with a non-trained panel.

**Results:** The values for the physicochemical characterization of the orange bagasse flour were in agreement with the literature, being the moisture content of 10%, which increases its shelf life and prevents deterioration. Fat content was low, less than 1% and ash and protein were less than 5% each. Muffin was evaluated and had the characteristics of a flour muffin, and after the sensorial profile, the panel showed a high percentage of acceptance of the product.

**Conclusions:** The use of natural products, like the orange bagasse, is a new way of using agro-industrial wastes in the food industry, in order to obtain a food product, with high added value, targeting a population with special needs such as celiac people.

**Keywords:** Orange bagasse, Gluten free muffin, Orange bagasse flour, Celiac, Functional food.

# EFFECTS OF THE PARTICLE SIZE OF YOUNG BARELEY LEAF POWDER ON INTESTINAL BACTERIAL FLORA IN RATS

(¹)Shimada, Ryoko; (²)Katayama, Masayuki; (³)Takano, Akira;

(⁴)Sugawa-Katayama, Yohko.

(¹)PhD. Department of Health and Nutrition. Osaka Aoyama University. Monoo. Japan.; (²)PhD. Visiting Professor. Department of Health and Nutrition. Osaka Aoyama University. Monoo. Japan.; (³)PhD. Research and Development Division, TOYO SHINYAKU Co., Ltd. Tosu. Japan; (⁴)PhD. Professor aemeritus. Department of Health and Nutrition. Osaka Aoyama University. Monoo. Japan.

**Background and objectives:** Young barley leaf powder (YBLP) is a component of the popular drink “Aojiru” used as a dietary supplement in Japan. We examined effects of the particle size of YBLP on the patterns of intestinal bacterial flora and fermented products in rats.

**Methods:** Seven week-old, male Wistar rats were divided into 4 groups (each 7 rats), and fed ad libitum for 14 days on the following AIN93G-based diets containing no cellulose (“fiber-free”), 4.5% cellulose (“C-4.5”), 3% cellulose plus 3% YBLP (fine particles) (“fine YBLP”), and 3% cellulose plus 3% YBLP (coarse particles) (“coarse YBLP”), respectively. Then, the rats were anesthetized with pentobarbital, and their ceca were removed and weighed. The short chain fatty acids of the cecal contents were determined by HPLC (High Performance Liquid Chromatography), and the patterns of intestinal bacteria flora were analyzed by T-RFLP (Terminal Restriction Fragment Length Polymorphism).

**Results:** The total amounts of the short chain fatty acids in the cecal contents of the “fine YBLP” and “coarse YBLP” groups were significantly greater than those of the “C-4.5” group. The concentration of n-butyric acid of the “fine YBLP” group was greater than that of the “coarse YBLP”, and the value was lowest in the “C-4.5” group. As for the intestinal bacterial flora, the ratio of Clostridiales species increased in the “fine YBLP” and in “coarse YBLP” groups while Bacteroidales increased in the “fine YBLP” group.

**Conclusions:** YBLP is suggested to contain some components favored by Bacteroidales, and the “fine YBLP” may show more clearly the efficacy of YBLP than the “coarse YBLP”.

**Keywords:** Young barley leaf powder, Intestinal bacterial flora, Short chain fatty acids.
EFFECTS OF PEPTIDES FROM JAPANESE BARLEY ON PC12 CELLS GROWTH AND MICROTUBULE FORMING

**Background and objectives:** Barley is useful as a foodstuff, because it contains high levels of protein, dietary fibers and minerals. We have searched the biological active peptides from Japanese barley flour obtained from the outer layer of the grain. Microtubules are important components of several subcellular structures. Recent reports have demonstrated that impairment of the microtubule system is related to the pathogenesis of Alzheimer's disease. Brain atrophy and decrease in cell number also have observed in Alzheimer's disease. In the present study, we investigated the effects of peptides obtained from barley flour on PC12 cells growth and microtubule assembly.

**Methods:** Japanese barley flour was extracted in water and the extract was dialyzed with a Cell Sep T1 (nominal MW=3,500). The external solution of dialysis was evaporated and taken to dryness. Then the sample was dissolved in water and subjected to size exclusion chromatography on a Bio Gel P-4 column. The molecular weights of these fractions were confirmed by HPLC analysis with Tosoh TSK 3000SW column.

PC12 cells were cultured in DME medium containing 5% FBS, and 0.3% penicillin/streptomycin and incubated at 37°in a 5% CO2 in a humidified atmosphere. After 48 h of incubation, the medium was replaced in with a fresh DME medium containing 0-10mg/ml P4-1 fraction. After 24h cultivation, cell viability was assessed using Cell Counting Kit-8. Percentage viability of cells in comparison to control cells was calculated. The GTPase assay was performed by HPLC system.

**Results:** Five fractions (P4-1, P4-2-1, P4-2-2, P4-3, and P4-4) were identified by the preceding sequential steps. From the results of analysis of molecular weight, P4-1 fraction was considered short peptides with four amino acids. Percentage viability of PC12 cells was increased depending on the levels of P4-1 fraction added.

**Conclusions:** Fraction P4-1, peptide from Japanese barley flour, is considered as PC12 cell growth factor. This oligopeptides promoted the microtubule assembly in PC12cells.

**Keywords:** Peptides, Barley, PC12 cells.

A NOVEL A-GLUCAN FROM AN EDIBLE MUSHROOM, GRIFOLA FRONDOSA (MAITAKE), EXERTS THE THERAPEUTIC EFFECTS ON INFLUENZA A VIRUS INFECTION

**Background and objectives:** Grifola frondosa (Fr.) S.F. Gray, called Maitake in Japanese, is an edible mushroom that has been consumed in Asian countries. We have isolated efficiently a novel α-glucan with MW of approximately 1 x 107 Da (Maitake α-glucan) from its fruiting body. In the present study, the therapeutic effects of the α-glucan were evaluated in immunocompetent and immunocompromised mice infected influenza A virus (IFV).

**Methods:** Maitake α-glucan was isolated from the dried materials by extracting with hot water and precipitating with ethanol. Oseltamivir-sensitive (A/NWS/33) and -resistant viruses (H1N1pdm09) were propagated on MDCK cells. In animal experiments, female BALB/c mice with normal immunity or immunocompromised mice treated with 5-fluorouracil were intranasally infected with IFV, and the α-glucan or oseltamivir was administered orally. Virus yields of the mice were measured at 3 days post-infection, neutralizing antibody titers were determined by plaque assays and virus-specific IgA by ELISA at 14 days post-infection.

**Results:** When the immunocompetent and immunocompromised mice were infected with an oseltamivir-sensitive IFV, the α-glucan suppressed the weight loss of the mice caused by virus infection and the virus replication, while increasing the neutralizing antibody and secretory IgA levels in both mouse groups. In the mice infected with oseltamivir-resistant IFV, the α-glucan suppressed the viral replication more markedly and produced higher titers of virus-specific antibody as compared with oseltamivir.

**Conclusions:** The present results suggested that Maitake α-glucan might be useful for therapy and prevention of IFV infections even in the immunosuppressive state.

**Keywords:** Maitake α-glucan, Influenza virus infection, Animal experiments, Therapeutic effects.

<footnote><sup>(1)</sup>Kawaguchi, Makiko; <sup>(2)</sup>Yamanaka-Nakayama, Yukako;</footnote>

<footnote><sup>(3)</sup>Takenouchi, Akiko; <sup>(4)</sup>Doi, Hiroshi.</footnote>

<footnote><sup>(1)</sup>PhD. Department of Food Design. Koshien University. Japan;</footnote>

<footnote><sup>(2)</sup>Department of Food Science and Nutrition. Kobe Shoin Women's University. Japan; <sup>(3)</sup>Department of Food Science and Nutrition. Mukogawa Women's University. Japan; <sup>(4)</sup>PhD. Department of Food Science and Nutrition. Mukogawa Women's University. Japan.</footnote>

<footnote><sup>(1)</sup>Hayashi, Kyoko; <sup>(2)</sup>Tanaka, Akihiro; <sup>(3)</sup>Lee, Jung-Bum; <sup>(1)</sup>Kawahara, Toshio; <sup>(1)</sup>Hayashi, Toshimitsu.</footnote>

<footnote><sup>(1)</sup>PhD. Research Institute of Life and Health Sciences. Chubu University. Japan.; <sup>(2)</sup>PhD. Yukiguni Maitake Co. Ltd. Japan.; <sup>(3)</sup>PhD. Graduate School of Medicine and Pharmaceutical Sciences for Research. University of Toyama. Japan.</footnote>
# BENEFICIAL EFFECTS OF SMALL MOLECULE OLIGOPEPTIDES ISOLATED FROM PANAX GINSENG C. A. MEYER IN LOW-DOSE ALLOXAN- AND HIGH-CARBOHYDRATE/HIGH-FAT DIET-INDUCED DIABETIC RATS, PARTIALLY BY AMELIORATE PANCR

Li, Yong.

Peking University.

**Background and objectives:** This study aimed at investigating whether treatment with oligopeptides from ginseng (GOP) could modulate type 2 diabetes mellitus (T2DM)-related hyperglycemia in rats induced by high fat diet and low doses of alloxan and its therapeutic mechanisms.

**Methods:** Male Sprague-Dawley rats made type 2 diabetic with 1 injections of 105 mg/kg alloxan and 4 weeks of the high-carbohydrate/high-fat diet were fed high-carbohydrate/high-fat diet with or without GOPs administration (0.125 g/kgBW, 0.5 g/kgBW and 2.0 g/kgBW) for 7, 24 and 52 weeks, respectively. Oral glucose test tolerance (OGTT), plasma glucose, serum insulin, the level of antioxidant and the score of b-cell function were measured. Morphological observation was performed by light microscopic analyses. The level of insulin and the expression of NF-κB signaling in pancreatic islets were also studied using western blot. Additionally, the survival time and rate were observed.

**Results:** After the treatment, the score of b-cell function and the abnormal OGTT of diabetic rats were partially reversed by GOPs treatment. The efficacious effect of GOPs was also manifested in the amelioration of pancreatic damage by microscopic analyses. Moreover, GOPs treatment increased normal insulin content and decreased the expression of NF-κB signaling and the number of apoptotic cells in diabetic islets. Additionally, compared with the model control, the survival time and rate were significantly longer.

**Conclusions:** These findings suggest that GOPs might have auxiliary therapeutic potential for diabetes, partially by ameliorate pancreatic beta-cell dysfunction and death.

**Keywords:** Panax ginseng oligopeptide, Pancreatic beta-cell failure, Type 2 diabetes mellitus, High--carbohydrate/high-fat, Alloxan.

**Further collaborators:** Some issue about T2DM

# EFFECT OF BLACK TEA DERIVED POLYPHENOLS ON MUSCLE ATROPHY AND RECOVERY IN MICE

(1)Aoki, Yuki; (2)Ozawa, Tetsuo; (1)Takemasa, Tohru; (2)Numata, Osamu.

(1)Graduate school of comprehensive human sciences, University of Tsukuba; (2)Graduate school of life and environment sciences, University of Tsukuba.

**Background and objectives:** Recently, there are some reports that polyphenols have some good effects on muscle. Our

laboratory originally purified high-molecular-weight polyphenols extracted from black tea. We named it Mitochondria Activation Factor (hereafter, MAF), because MAF enhanced mitochondrial membrane potential of ciliated protista Tetrahymena pyriformis (Fujihara et al.). We found MAF raises AMPK (regulator of PGC-1 alpha, which stimulates mitochondria biogenesis) phosphorylation level and improves endurance performance in mice (Eguchi et al.). Recently, we extracted crude MAF fraction with 80% ethanol, and named this fraction E80. In this study, we examined whether E80 prevented skeletal muscle of mice during tail suspension or promoted recovery from muscle atrophy.

**Methods:** Adult mice (8 weeks of age) were fed on food containing 0.5% E80 during 2 weeks of tail suspension and 5 days or 10 days of recovery after 2 weeks of tail suspension. After these process, mice were euthanized by cervical dislocation and excised soleus muscle.

**Results:** We measured muscle wet weight and molecules regulating muscle mass. Then, we found E80 may have potential for promoting recovery of muscle mass from atrophy by tail suspension.

**Conclusions:** Previously, we reported E80 promote muscles hypertrophy during functional overload in mice. In this study, we demonstrated E80 has potential for promoting recovery of muscle mass from atrophy.

**Keywords:** Polyphenol, Unloading, Reloading, Skeletal, Muscle.

# RESVERATROL AMELIORATES RENAL ISCHEMIA-REPERFUSION INJURY VIA ACTIVATING GLYOXALASE 1 ACTIVITY AND THIOREDOXIN REDUCTASE/THIOREDOXIN SYSTEM

(1)Yeh, Wan Ju; (2)Pai, Man-Hui; (3)Tan, Hsiu-Yun; (1)Chen, Jiun-Rong;

(4)YANG, HSIN-YI.

(1)PhD. School of Nutrition and Health Sciences. Taipei Medical University. Taipei. Taiwan.; (2)PhD. Department of Anatomy and Cell Biology. School of Medicine. College of Medicine. Taipei Medical University. Taipei. Taiwan.; (3)MSc. School of Nutrition and Health Sciences. Taipei Medical University. Taipei. Taiwan.

(4)PhD. Department of Nutrition. I-Shou University. Kaohsiung. Taiwan.

**Background and objectives:** The glycative stress is partially responsible for the damage of renal ischemia-reperfusion (IR) injury. Accumulating evidences indicate that resveratrol has beneficial effects on markers of oxidative stress in diseases. In this study, we used a renal IR model to demonstrate pretreatment of resveratrol showed the protective effects on renal IR injury.

**Methods:** Mice were assigned to sham-operation group, renal IR group, pretreat apocynin, and pretreat resveratrol group. Serum cystatin C and creatinine (Cr) levels were analyzed. The renal methylglyoxal (MG), malondialdehyde (MDA) levels and also glyoxalase 1 (GLO-1) activity were detected. Renal thioredoxin in reductase/thioredoxin system and caspase 3 were measured by western blotting.
**Results:** Our model of renal IR resulted in severe kidney dysfunction 24 hour after reperfusion indicated by significant increases in serum Cr levels. We also determined that mice subjected to IR increased MG accumulation and MDA level and decreased GLO-1 activity in the kidneys. Compared with vehicle control, intravenous tail vein injection of resveratrol (10 mg/kg bw) markedly improved the renal function, reduced oxidative stress, and cell apoptosis in renal IR injury models, strongly suppressed the IR-induced endogenous MG accumulation and increased the activity of GLO-1.

**Conclusions:** This study suggests that pretreatment of resveratrol could reduce the burden of endogenous MG accumulation-induced by IR with renal impairment and possibly improve the severity of renal injury.

**Keywords:** Resveratrol, Renal ischemia-reperfusion injury, Glyoxalase 1, Methylglyoxal.

144/1113

PROTECTIVE ACTION OF PHELLINUS LINTEUS POLYSCCHARIDE AGAINST RADIATION-INDUCED HAEMATOPOIETIC ABNORMALITY IN MICE

(1)Huang, Shu-Ming; (2)Chen, Chin Chu; (3)Hu, Miao Lin.

(1)PhD. Department of nutrition. Nantou Hospital. Ministry of Health and Welfare. Nantou City of Taiwan. R.O.C.; (2)PhD. Biotechnology Center. Grape King Inc. Taoyuan. Taiwan. R.O.C.;

(3)PhD. Agricultural Biotechnology Center. National Chung Hsing University. Taichung Taiwan. R.O.C.

**Background and objectives:** Radiation exposure is one of the potential carcinogenic factors in our living environment. The hematopoietic system damage is the main side effect of radiation exposure. Phellinus linteus (PL), one of the medicinal fungus in East Asia, contains rich polysaccharides, flavonoids and polyphenols, been shown to exhibit anti-cancer, anti-inflammatory and immunomodulative effects. We determined the protective effect of PL polysaccharide (PLP) on low dose of irradiated mice

**Methods:** Mice were divided into control group (no irradiation), irradiated group (2 Gy), irradiated (2 Gy) + WR2721 (200 mg / kg), irradiated (2 Gy) + low dose group (100 mg / Kg), irradiated (2 Gy) + high dose (500 mg / kg). Blood samples were measured 48 hours after the first irradiation. WBC, RBC, HGB, PLT. Blood samples were collected for 48 hours after each irradiation. The weight was measured weekly and recorded. In the sixth week of sacrifice. Sacrifice after the collection of blood and mouse organs, intestinal, femur for analysis.

**Results:** Results revealed that PLP administration, especially at high dose, significantly restored radiation-induced loss of WBC and platelets. In addition, PLP supplementation significantly ameliorated radiation-induced femur fibrosis. Similarly, PLP effectively reversed radiation-induced abnormal serum levels of immunological parameters.

**Conclusions:** Overall, this dissertation research demonstrated that PLP effectively protected against radiation damage by increasing antioxidant activities and ameliorating abnormal hematopoietic in mice.

**Keywords:** Radiation, Phellinus Linteus, Polysaccharides, Hematopoietic function.

144/1152

PRELIMINARY STUDIES OF ELABORATION, ACCEPTABILITY AND NUTRITIONAL QUALITY OF FORMULATIONS RICH IN OMEGA-3 FATTY ACIDS FOR THEIR USE IN EXPERIMENTAL MURINE MODELS

(1)Colussi, Narella Antonina; (2)Romero Vidomlansky, Patricia Ruth;

(3)Todaro, Juan Santiago; (4)Melana Colavita, Juan Pablo; (5)Aguirre, Julio Andrés; (6)Gorbán, Silvia Beatriz; (7)Aguirre, María Victoria.

(1)MSc. Degree in Nutrition. Biochemical Research Laboratory. Faculty of Medicine. National Northeastern University (UNNE). Corrientes. Argentina.; (2)Food Engineer. Biochemical Research Laboratory. Faculty of Medicine. National Northeastern University (UNNE). Corrientes. Argentina.; (3)Degree in Biology. Biochemical Research Laboratory. Faculty of Medicine. National Northeastern University (UNNE). Corrientes. Argentina.;

(4)Biochemist. Biochemical Research Laboratory. Faculty of Medicine. National Northeastern University (UNNE). Corrientes. Argentina.; (5)Under Graduate Student. Medicine Career. Faculty of Medicine. National Northeastern University (UNNE). Corrientes. Argentina.; (6)MD. Nutrition Professor. Kinesiology and Physical Education Career. Faculty of Medicine. National Northeastern University (UNNE). Corrientes. Argentina.; (7)PhD. Biochemist. Biochemistry Professor. Biochemical Research Laboratory. Faculty of Medicine. National Northeastern University (UNNE). Corrientes. Argentina.

**Background and objectives:** Background: Basic metabolic research of chronic non transmissible diseases often needs the use of different diets in experimental animals, these must be designed for being complete. Given the already observed functional effects of Omega-3 fatty acids and their high content in chia seeds, it is interesting to find a suitable method for their incorporation into the murine diet. Objective: to evaluate the organoleptic characteristics, acceptability and nutritional quality of food rich in Omega-3 fatty acids in Balb-c mice from the Biotherium of Faculty of Medicine, Universidad Nacional del Nordeste.

**Methods:** Adult mice (2 months) of the Balb-c strain were used. The protocol was approved by the Institutional Committee for Care and Use of Experimental Animals. Mice were randomly distributed in 4 lots with food and water ad-libitum. Experimental groups received a diet rich in Omega-3, while the control lots (A) were provided with a standard commercial feed for 15 days. The diets were prepared manually with natural ingredients according to nutritional requirements: one formulation with egg proteins (B) and one with milk and egg (C). The semi-solid preparations were
molded as standard pellets, which were: 1) dried in an oven for 2 h at 80°C, and 2) autoclaved (103 kPa, 120°C) 10 min and further dried 5 h at 40°C. Thus, the four diets were: B1, B2, C1, C2. The evaluation of diets include: the organoleptic characteristics of the pellets, its acceptability and digestibility. Measurements involved the progression of mice body weight, the consumption of pellets and the study of feces. Coprologic samples were analyzed with light microscopy (40x).

**Results:** The smell and color of the food were conventional and similar among different groups. The diet with the greatest acceptability was C. Animals fed with diet B decreased body weight in comparison with those fed with diet C, which maintained or increased it similarly to controls.

**Conclusions:** This study let us conclude that the best formulation of a pellet rich in Omega-3 from chia seeds is C1. Additionally, this work might be used for evaluating Omega-3 bioavailability in further settings for metabolic models of chronic non transmissible diseases.

**Keywords:** Functional food, Nutrition, Chia seeds, Food technology.

**Conclusions:** Our results suggested that PE consumption may improve ethanol-induced hepatic steatosis and inflammation, and may be beneficial in ameliorating the progression of alcoholic liver diseases.

**Keywords:** Pitaya, Ethanol, Lipid, Liver, Inflammation.

**Background and objectives:** In recent years mankind has produced the most significant changes in their eating patterns and lifestyle. Just as hunger and malnutrition were the main focus of food policy for decades, malnutrition, anemia, and obesity are today. Increased consumption of saturated fats, simple carbohydrates and decreased fiber intake leads to a higher incidence of chronic noncommunicable diseases (CNCDs). The WHO proposes that industries produce healthier foods.

The objectives of the work were to elaborate a cereal bar fortified with iron, fiber and antioxidants and to determine its physicochemical properties, microbiological stability and sensorial acceptability.

**Methods:** A cereal bar was made with oats, oat bran, inflated corn, honey, fortified sugar with iron, lemon zest and sunflower oil. Variation and physical chemical and microbiological stability were studied under different storage conditions. The sensorial properties and the acceptability were evaluated by means of the hedonic scale in a population of 100 people between 9 and 78 years of both sexes residing in Tucumán.

**Results:** The formulation of the cereal bar is made up of 52% carbohydrates, 12% protein and 36% fat. The values of fiber (1.99 grs) and iron (0.4 mg) position it as a functional food.

With respect to acceptability, 96% of the participants considered it excellent and this facilitates their introduction into different age groups.

The study of the physical and chemical characteristics of the product, are stable being the best method to keep the dark wrapping.

**Conclusions:** Conclusion, a food rich in iron and with antioxidant properties was obtained. The physical and chemical characteristics of the product are stable being the best method to keep the dark packaging. With respect to acceptability, more than half of the participants considered it excellent and this facilitates their introduction into different age groups.

According to scientific evidence fiber and antioxidants have positive effects on CNCDs and iron fortification prevents anemia in schoolchildren and pregnant women. It is proposed to bring to the market a cereal bar with the mentioned compounds.

**Keywords:** Functional food, Cereal bar, Nutrition, Health.

**Further collaborators:**

Mg. Ing. Patricia Albarracín, Dra. Norma Barnes, Dra. Dora Paz, Dra. Martha Kairuz.

**Background and objectives:** Alcohol consumption leads to liver injury, ectopic fat accumulation in the liver and excretion of proinflammatory cytokines which may promote the progression of liver disease. Red pitaya is one of the popular fruit in Taiwan and its disposed peel is rich in phenolic compounds. Thus, the aim of the study is to investigate the beneficial effects of pitaya peel extract (PE) in a chronic plus single binge ethanol feeding model.

**Methods:** Twenty-four C57BL/6 mice were assigned to three groups consuming control liquid diet (C), ethanol liquid diet (E) or ethanol liquid diet plus 1000mg/kgBW red pitaya peel extract (P) for 13 days. At day 14, mice were orally gavaged with a single dose of isocaloric maltodextrin (C) or ethanol (5g/kgBW, E & P). After 9h, mice were sacrificed and we collected their blood and liver tissue for analysis. In addition, 20 rats were assigned to two groups to clarify whether PE affected ethanol absorption.

**Results:** At the end of the study, we found that ethanol diet significantly elevated serum and hepatic lipid concentration and increased the hepatic malondialdehyde (MDA) and proinflammatory cytokines while PE consuming group had lower hepatic lipid, MDA and interleukin (IL)-6 concentrations than the E group. We also found that PE consumption did not directly affecting the ethanol absorption rate.
# EFFECT OF THE CONSUMPTION OF HAMBURGERS PREPARED WITH ADDITION OF WINE GRAPE POMACE FLOUR, RICH IN FIBER AND ANTIOXIDANTS, ON PLASMA ANTIOXIDANTS

(1)Perez, Druso; (2)Urquiaga, Ines; (3)Dicenta, Sara; (4)de la Cerda, Paula; (5)Urzua, Catalina; (6)Troncoso, Danitza; (6)Mackenna, Maria Jose; (7)ECHEVERRÍA, GUADALUPE; (8)Rigotti, Attilio.

(1)MSc.; (2)PhD. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile.; (3)MSc. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile.; (4)Biochemist. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile.; (5)Food Engineering. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile.; (6)MD. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile; (7)MSc. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile; (8)PhD. Centro de Nutricion Molecular y Enfermedades Crónicas. Departamento de Nutrición, Diabetes y Metabolismo, Escuela de Medicina. Pontificia Universidad Católica de Chile.

**Background and objectives:** Diet is a modifiable factor for prevention of chronic diseases. Antioxidants and fiber are two bioactive components with proven beneficial health properties. Currently, fiber and antioxidants population intake is less than that recommended by the WHO. The objective of this study was to develop a functional food, enriching a high consumption meat-based food with wine grape pomace flour (WGPF), rich in fiber and antioxidants, and to study the effect of its consumption on plasma antioxidants in an intervention study.

**Methods:** We formulated a beef hamburger (100 g) with 7% of WGPF, containing 3.5% of fiber, 1.2 mg EAG/g of polyphenols and 17.2 µmoles Trolox/g of ORAC (WGPF-hamburger). WGPF was prepared from pressed of grapes after winemaking (Caber-net Sauvignon). A controlled intervention study of 3 months was completed in 27 male volunteers. Volunteers daily consumed one WGPF-hamburger during the first month, interrupted by a month washout period, followed by a control month period where they daily consumed a hamburger without WGPF (control-hamburger). Plasma antioxidants (vitamin C, vitamin E, beta-carotene and antioxidant capacity as DPPH) were evaluated at time 0, 1, 2 and 3 months.

**Results:** Intake of the WGPF-hamburger significantly increased vitamin C (p<0.001), whereas intake of control-hamburger significantly decreased vitamin C (p=0.005), vitamin E (0.050) and beta-carotene (p=0.040). No significant effect was observed in plasma antioxidant capacity during WGPF-hamburger and control-hamburger periods.

**Conclusions:** The consumption of beef hamburger elaborated with 7% WGPF protects against plasma antioxidant expenditure, vitamin C, vitamin E and beta-carotene and enhances vitamin C, showing its potential to prevent oxidative damage and the functionality of this food.

**Keywords:** Antioxidant, Oxidative damage, Fiber, Functional food.

# MACAÍBA (ACROCOMIA INTUMESCENS DRUDE) DECREASED VISCERAL FAT IN DISLIPIDEMIC RATS

(1)Albuquerque de Souza, Mikaelle; (2)Tavares-de-Melo, Marília;

(3)Melo, Flávio; (4)Silva, Jaielison; (4)Oliveira, Shirlayne; (4)Ponciano,

Carolina; (4)Silva, Roberta; (5)Bertozzo, Camila; (6)Oliveira, Maria

Elieidy; (6)Soares, Juliana.

(1)MsC. Nutritionist. Universidade Federal da Paraíba.; (2)MsC.

Nutritionist. Universidade Federal de Campina Grande.;

(3)Pediatrician. Hospital Regional de Guarabira. Instituto Cândida

Vargas. Círculo do Coração de Pernambuco.; (4)Nutrition Student.

Universidade Federal de Campina Grande.; (5)PhD. Pharmacist.

Universidade Federal de Campina Grande.; (6)PhD. Nutritionist.

Universidade Federal de Campina Grande.

**Background and objectives:** Macaíba is a palm tree. It has significant percentages of unsaturated fatty acids in its composition, such as oleic and linoleic acid. In addition, it is a source of vitamin as well as high levels of carotenoids and vitamin C. The objective of this work was to evaluate the effects of macaíba pulp intake on murinometric parameters and visceral fat of dyslipidemic adult Wistar rats.

**Methods:** Male rats were divided into four groups: Control (CT), Dyslipidemic (DL), Macaíba (MC) and Dyslipidemic Macaíba (MCDL), using n=9 per group. Dyslipidemia was previously induced during 14 days. The animals received 1000mg/ kg of body weight/day of macaíba pulp by gavage during 28 days. The research was conducted according to the Guidelines for the Care and Use of Experimental Animals and was submitted and approved by the Ethics Committee on Animal Use (CEUA) of the Center for Health and Rural Technology - CSTR / UFCG, protocol number CEP 063 / 2016. Data were evaluated with One Way Anova followed by Tukey (p<0.05) using Sigma Stat software.

**Results:** Feed intake, abdominal and thoracic circumference, weight, length, BMI and visceral fat were evaluated. The data demonstrated that there was no difference in the physical parameters, but a reduction of visceral fat in MC and MCDL was verified. Studies show that monounsaturated fats reduce the amount of circulating fats, and therefore fat uptake by adipocytes is reduced, thus reducing visceral fats is associated with an increased risk of developing heart disease. Therefore, macaíba shows to be a promising food for preventing and/or combatting lipidic alterations that may promote correlated pathologies such as dyslipidemia.

**Conclusions:** Therefore, macaíba shows to be a promising food for preventing and/or combatting lipidic alterations that may promote correlated pathologies such as dyslipidemia.

**Keywords:** Visceral fat. Acrocomia intumescens Drude. Dyslipidemia.
## BIOCHEMICAL PARAMETERS OF DISLIPIDEMIC RATS CONSUMING MACAÍBA (ACROCOMIA INTUMESCENS DRUDE)

**Background and objectives:** Macaíba is a palm tree that can be found in the northeastern and southeastern regions of Brazil as part of the Atlantic Forest Biome. The pulp stands out as a source of fiber, fatty acids, such as oleic and linoleic acid and essential aminoacids. In addition, it is a source of vitamin C and contains significant levels of total carotenoids and yellow flavonoids in its composition. Natural antioxidants such as carotenoids, vitamin C, and unsaturated fatty acids present in fruits, greenery, and vegetables have been subject of study in the fight against changes in blood lipid levels, associated with the risk of developing cardiovascular diseases. The objective of this work is to evaluate the effects of macaíba pulp intake on biochemical parameters of dyslipidemic adult Wistar rats.

**Methods:** Male rats were divided into four groups: Control (CT), Dyslipidemic (DL), Macaíba (MC) and Dyslipidemic Macaíba (MCDL), using n=9 per group. Dyslipidemia was previously induced during 14 days. The animals received 1000mg/kg of body weight/day of macaíba pulp by gavage during 28 days. Data were evaluated with One Way Anova followed by Tukey (p<0.05) using Sigma Stat software. Oxalacetic glutamic transaminase (OGT), pyruvic glutamic transaminase (PGT), creatinine, urea, total cholesterol (TC), triglycerides (TG), low density lipoprotein (LDL), high density lipoprotein (HDL) and atherogenic risk were calculated.

**Results:** The data demonstrated that there was a reduction of all the biochemical parameters in MCDL and TG in MC. MC and MCDL groups presented increases in HDL and decreases in atherogenic index (p<0.05). MCDL also presented a reduction in PGT and OGT.

**Conclusions:** Therefore, macaíba shows to be a promising food for preventing and/or combatting lipidic alterations that may promote correlated pathologies such as dyslipidemia.

**Keywords:** Dyslipidemia. Acrocomia intumescens Drude. Rat.

## ORAL ADMINISTRATION OF FREEZE-DRIED POWDERS OF HONEYBEE LARVAE INHIBITS DEVELOPMENT OF ATOPIC DERMATITIS-LIKE SKIN LESIONS IN NC/NGA MICE

**Background and objectives:** Honeybee larvae was pupa of honeybees and has been eaten since ancient days because human being expects/expected of its high nutrient with rich amino acid score. The suppressive effect of the oral administration of honeybee larvae on the atopic dermatitis (AD)-like skin lesions in NC/ Nga mice was evaluated.

**Methods:** NC/Nga mice are used as the most extensively studied animal model of AD.

**Results:** Oral administration of 0.5% and 1.0% honeybee larvae-added diet for 46 days inhibit the development of AD-like skin lesions, and elevation of serum IgE levels, accompanying with the declined serum levels of both IL-4 and IFN-γ and IL-18 and IL-12 contents in the skin lesions. These results suggest that honeybee larvae inhibits the development of skin lesions in NC/Nga mice by suppressing both the T-helper (Th) 1 and Th 2 cell responses.

**Conclusions:** Our results indicate that oral administration of honeybee larvae could provide an adjuvant therapy for the management of AD.

**Keywords:** Atopic dermatitis, Honeybee larvae, Interferon-γ, Interleukin-4, NC/Nga mice.

## THE EFFECTS OF ALMOND (PRUNUS DULCIS) ON THE POSTPRANDIAL BLOOD GLUCOSE LEVELS IN MICE

Kato, Kazuko; Vo, Phuong; Furuyashiki, Takashi; Kamasaka, Hiroshi; Kuriki, Takashi.

Ezaki Glico Co., Ltd. Institute of Health Sciences. Osaka. Japan

**Background and objectives:** Almond is generally thought to be effective on maintaining and enhancing our health. Recently some reports revealed that almond have an effect for decreasing the postprandial blood glucose level when carbohydrate and al-

<footnote>(1) Albuquerque de Souza, Mikaelle; (2) Tavares-de-Melo, Marília;</footnote>

<footnote>(3) Melo, Flávio; (4) Silva, Jaielison; (4) Oliveira, Shirlayne; (4) Ponciano, Carolina; (4) Silva, Roberta; (5) Bertozzo, Camila; (6) Oliveira, Maria Elieidy; (6) Soares, Juliana.</footnote>

<footnote>(1) MsC. Nutritionist. Universidade Federal da Paraíba.; (2) MsC. Nutritionist. Universidade Federal de Campina Grande.;</footnote>

<footnote>(3) Pediatrician. Hospital Regional de Guarabira. Instituto Cândida Vargas. Círculo do Coração de Pernambuco.; (4) Nutrition Student. Universidade Federal de Campina Grande.; (5) PhD. Pharmacist. Universidade Federal de Campina Grande.; (6) PhD. Nutritionist. Universidade Federal de Campina Grande.</footnote>

<footnote>(1) Kageyama, Masakatsu; (2) Xing, Guoqing; (3) Li, Kejuan; (4) Zhang, Zhenya; (5) Sugiyama, Akihiko.</footnote>

<footnote>(1) Msc. Division 2. DHC Corporation Laboratories. Chiba Prif. Graduate School of Life and Environmental Sciences. Tsukuba University. Ibaraki Pref. Japan.; (2) PhD. Researcher. Nature Industry of Tsukuba Co., Ltd. Japan.; (3) PhD. Researcher. Graduate school of life and Environmental Sciences. Tsukuba University. Japan.; (4) PhD. Professor. Graduate school of life and Environmental Sciences. Tsukuba University. Japan.; (5) PhD. Associate professor. Joint Department of Veterinary Medicine, Faculty of Agriculture. Tottori University. Japan.</footnote>
mond are ingested simultaneously in human study. However the detail of the effect including functional components and mechanism have still been unclear. In this study, we assessed the functional components and mechanism of the effect.

**Methods:** After being fasted for overnight, male ICR mice were orally administrated maltodextrin with one of the following diet, depending on the group; (1) whole almond paste (4 g/ kg), (2) almond paste without seed coat (4 g/kg), or each fraction fractionated from whole almond; (3) almond oil, (4) water soluble fraction, (5) water insoluble fraction, (6) water as a control. The blood samples were collected from the tail at the time of 0, 15, 30, 45, 60, 90, 120 min after the administration and applied for the measurements of blood glucose and insulin level. To compare the effects of oils, we also tested the effects of other kinds of oils, that is, peanut oil, walnut oil and olive oil.

**Results:** The mice ingested maltodextrin with almond paste (with and without seed coat) showed significantly lower value of the blood glucose level and higher of the insulin level. Although the soluble and insoluble fractions did not show any effects, only almond oil significantly decreased the postprandial blood glucose level without the increase of insulin level, compared to the control group. Comparing the effect of oils, the walnut and olive oils showed the same tendency as that of almond oil, but peanuts oil did not.

**Conclusions:** Whole almond suppresses the increase of postprandial blood glucose level via enhancement of insulin secretion. Almond oil also decreased the increase of blood glucose without insulin spike.

**Keywords:** Almond, Blood glucose, Insulin, Oil, Mouse.

ly correlated with plasma glucose levels and insulin resistance in KK-Ay mice. In the present study, we have attempted to examine antidiabetic and antihyperuricemic effects of TXF in cultured L6 myotubes and KK-Ay mice.

**Methods:** L6 myoblasts were cultured in 10% fetal bovine serum (FBS)/ Dulbecco's Modified Eagle Medium (DMEM) and differentiated to L6 myotubes in 2% FBS/DMEM. Then, glucose uptake assay was conducted as already mentioned by us (Kawano et al. 2009) in the absence or presence of TXF. KK-Ay mice were maintained on regular pellet diet, and TXF (30 mg/kg body weight) suspended in vehicle (0.5% CMC-Na) was orally administered once a day for 4 weeks. Normal and diabetic control mice were given vehicle alone. At the end of experimental period, the blood was collected under isoflurane anesthesia from the inferior vena cava in the microtube with heparin to obtain plasma. Concentrations of plasma glucose, insulin and uric acid were determined with commercial kits.

**Results:** TXF dose-dependently (0-200 µM) and significantly promoted glucose uptake in L6 myotubes, and the rise was cancelled by Compound C (10 µM), an AMPK inhibitor, indicating an involvement of AMPK and hence GLUT4 translocation in TXF action. Plasma glucose, insulin, uric acid concentrations and HOMA-IR, an index of insulin resistance, significantly elevated in diabetic control KK-Ay mice as compared with normal ones. All the rises were significantly suppressed by TXF treatment.

**Conclusions:** TXF is demonstrated to be antidiabetic and antihyperuricemic.

**Keywords:** AMPK. Diabetes. Hyperglycemia. Hyperuricemia. KK-Ay mice.

144/1422

ANTIDIABETIC AND ANTIHYPERURICEMIC EFFECTS OF TAXIFOLIN, A POLYPHENOL PRESENT IN OCTAPLOID STRAWBERRY, IN CULTURED L6 MYOTUBES AND TYPE 2 DIABETIC MODEL KK-AY MICE

**Background and objectives:** The number of diabetic patients is increasing globally. Many trials to suppress hyperglycemia by use of phytochemicals have been conducted so far. The skeletal muscle is the largest tissue in our body and plays an important role in maintaining postprandial glucose homeostasis by uptake of blood glucose through glucose transporter 4 (GLUT4) via AMP-activated protein kinase (AMPK) phosphorylation. We have recently demonstrated that type 2 diabetic (T2D) model KK-Ay mice accompany hyperuricemia as well as hyperglycemia and that taxifolin (TXF) suppresses purine bodies-induced hyperuricemia. We have also found that plasma uric acid levels are positive-

144/1428

EFFECT OF THE CONSUMPTION OF EPA AND DHA DERIVED FROM MICROALGAE IN CD1 MICE

Martinez-Carrillo, Beatriz Elina; Valdes-Ramos, Roxana; Soto-Piña, Alexandra; Gutiérrez-Pliego, Laura Elisa; Escoto-Herrera, Jorge Alberto; Rosales-Gómez, Cristian.

Universidad Autónoma del Estado de México.

**Background and objectives:** The alpha-linolenic fatty acid (C18:3, ALA) belonging to the n-3 family, is an essential fatty acid, cannot be synthesized in the body and should be consumed to maintain health. EPA (ecosapentanoic acid) and DHA (docosahexaenoic acid) have vegetable sources (algae and microalgae) and animals (fish, crustaceans, seafood) of marine origin as a dietary source. Cultured microalgae, a renewable source with high EPA and DHA content, can be consumed lyophilized or microencapsulated and added to food, since they have great potential in human and animal nutrition. The objective is to know the effect of the consumption of EPA and DHA derived from microalgae in CD1 mice.

**Methods:** 32 db/db mice, 56 days old, divided into 4 groups (n = 8) were used: a) Control (CL), b) EPA and DHA lyophilisate (Li), c) Croquette + 2% EPA and DHA (Cr), d) Coconut oil (CO).

<footnote>(1)Yagasaki, Kazumi; (1)Kondo, Shinji; (1)Adachi, Shin-ichi;</footnote>

<footnote>(2)Yoshizawa, Fumiaki.</footnote>

<footnote>(1)PhD. Centre for Bioscience Research and Education.
Utsunomiya University. Japan.; (2)PhD. Faculty of Agriculture.
Utsunomiya University. Japan.</footnote>
Treatment was given for 8 weeks and sacrificed at day 112. BMI, Body weight, feed intake, water, glucose, total cholesterol, triglycerides and HDL were measured.

**Results:** No differences were found in BMI (p=0.117); feed intake was higher (p<0.002) in the CO group (66.05 g/day ± 2.1) and lower in the Li group (42.47 g/day ± 4.2); the water intake increased (p<0.001) in the CO group (86.02 ml/day ± 3.9) and decreased in the Li group (55.01 ml/day ± 2.3) compared to the control (67.27 ml/day ± 2.7). The concentration of glucose (p<0.008) increases with Li (127.3 mg/dL ± 5.7), but decreases with Cr (109.5 mg/dL ± 1.2) and CO (106.6 mg/dL ± 1.4). Total cholesterol did not change (p=0.062); HDL-cholest decreased (p<0.001) in the Cr group (39.2 mg/dL ± 8.4) compared to the control (50.25 mg/dL ± 9.8); triglycerides increasing (P <0.001) in the CO group (98.5 mg/dL ± 4.2), decreasing in the Li (60 mg/dL ± 2.4) and Cr (67.5 mg/dL ± 2.2) groups.

**Conclusions:** The effect of supplementation with EPA and DHA from cultured marine microalgae is beneficial; it does not modify BMI; the lyophilized decreases feed consumption and water; although increasing blood glucose levels, improves the concentration of triacylglycerols.

**Keywords:** Microalgae, Ecosapentanoic acid, Docosahexaenoic acid.

BMI, Body weight, feed intake, water, glucose, total cholesterol, triglycerides and HDL-colest were measured.

**Results:** No differences were found in BMI (p=0.248); feed intake (p<0.011) was higher in the CO group (36.12 g/w ± 1.85) and lower in the Cr group (28.82 g/w ± 2); the water intake increased (p<0.031) in the Li group (76.32 ml/day ± 4) compared to the control (69.73 ml/day ± 1.32). No differences were found in glucose concentration (p = 0.132); total cholesterol was elevated (p<0.015) in the Li and Cr groups; HDL-cholest increased (p<0.001) in the group that consumed Li (92.1 mg/dL ± 3.9), compared to its control (57.4 mg/dL ± 6); the triacylglycerols were modified (p<0.004), increasing in the Li group (356.2 mg/dL ± 27.4) and decreasing in the CO group (199.6 mg/dL ± 2.88).

**Conclusions:** In diabetic mice, the effect of supplementation with EPA and DHA is beneficial, as it does not modify BMI, decreases feed intake and does not alter blood glucose, further substantially improves the concentration of HDL-cholest.

**Keywords:** Microalgae, Ecosapentaenoic acid, Docosahexaenoic acid, Anthropometric parameters.

144/1430

EPA AND DHA CONSUMPTION EXTRACTED FROM MICROALGAE AND ITS EFFECT ON ANTHROPOMETRIC AND BIOCHEMICAL PARAMETERS

Martinez-Carrillo, Beatriz Elina; Valdes-Ramos, Roxana; García-Rillo, Arturo; Gutiérrez-Pliego, Laura Elisa; Escoto-Herrera, Jorge Alberto; Rosales-Gómez, Cristian.

Universidad Autónoma del Estado de México.

**Background and objectives:** Long chain polyunsaturated fatty acids (LC-PUFA) are dietary components that participate in multiple physiological processes; the alpha-linolenic fatty acid (C18:3, ALA) belongs to the n-3 family, cannot be synthesized in the body, a minimum consumption for health should be maintained. The ALA can be transformed into EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid), its dietary sources are vegetable organisms (algae and microalgae) and animals (fish, crustaceans, seafood) of marine origin. Cultured microalgae are a renewable source with high EPA and DHA content, can be consumed lyophilized or microencapsulated and added to a wide variety of foods as they have great potential in human and animal nutrition. The aim of this study was to determine the relationship between the consumption of EPA and DHA extracted from microalgae with anthropometric and biochemical parameters.

**Methods:** 32 db/db mice at 8 weeks of age, divided into 4 groups (n = 8): a) Control (CL), b) EPA and DHA lyophilisate (Li), c) Croquette + 2% EPA and DHA (Cr), d) Coconut oil (CO). They were given the treatment for 8 weeks and sacrificed at week 16.

144/1460

DIETARY AMINO ACIDS CONTROL LIPID ACCUMULATION IN THE LIVER, SKELETAL MUSCLE OR ADIPOSE TISSUE IN AN AMINO ACID-SPECIFIC MANNER

(1)Yamanaka, Daisuke; (2)Nishi, Hiroki; (2)Goda, Yuki; (2)Kumano, Mikako; (2)Taniguchi, Sakiko; (3)Ito, Koichi; (4)Katsumata, Masaya;

(5)Kato, Hisanori; (6)Hakuno, Fumihiko; (3)Takahashi, Shin-Ichiro.

(1)PhD. Assistant Professor. Department of Veterinary Medical Sciences. Graduate School of Agriculture and Life Sciences. The University of Tokyo. Japan; (2)Graduate student. Graduate School of Agricultural and Life Sciences. The University of Tokyo. Japan.;

(3)PhD. Associate Professor. Graduate School of Agricultural and Life Sciences. The University of Tokyo. Japan.; (4)PhD. Professor. School of Veterinary Medicine. Azabu University. Japan.; (5)PhD. Professor. Organization for Interdisciplinary Research Program. The University of Tokyo. Japan.; (6)PhD. Assistant Professor. Graduate School of Agricultural and Life Sciences. The University of Tokyo. Japan.

**Background and objectives:** We have previously shown that dietary protein deprivation or dietary arginine restriction induces accumulation of neutral lipid in the liver, and by contrast dietary lysine restriction resulted in lipid accumulation in the skeletal muscle. These studies suggest that not only amounts but also balance of amino acids control lipid accumulation in a tissue-specific manner. However, effects of a single amino acid on lipid accumulation in the peripheral tissues remain largely unclear, and unveiling these effects can be a clue to understand amino acid-monitoring systems. In this study, we aimed to elucidate effects of supplementation with a single amino acid in an amino acid-deficient diet on lipid accumulation in the peripheral tissues.
**Methods:** We fed six-week-old male Wistar rats for one week with diets containing sufficient (15%) amino acids (control diet) or insufficient (5%) amino acids (5AA diet), or 20 kinds of single amino acid-supplemented 5AA diets in which a single amino acid was increased to the same levels as the control diet, and then measured neutral lipid content in the liver and skeletal muscle or weight of adipose tissue.

**Results:** Triglyceride content in the liver was increased in a 5AA group compared to a control group. Feeding of methionine- or cystine-supplemented 5AA diets enhanced this increase. In the skeletal muscle (longissimus thoracis), triglyceride content in the 5AA group was comparable to that in the control group. However, supplementation of leucine or arginine in the 5AA diet increased triglyceride content in the muscle. Analysis of weight of adipose tissue (retroperitoneal fat) revealed that feeding the 5AA diet increased the adipose tissue weight compared to the control group. In addition, supplementation with serine in 5AA diet enhanced the increase in the adipose tissue weight.

**Conclusions:** In summary, methionine or cystine supplementation increased neutral lipid content in the liver, leucine or arginine increased it in the skeletal muscle, and serine increased the adipose tissue weight. Taken together with other results, we concluded that a single amino acid could control lipid accumulation in a tissue-specific manner in growing rats under conditions of dietary amino acid insufficiency.

**Keywords:** Amino acids, Fatty liver, Marbled muscle, Adipose tissue.

**Background and objectives:** Two Bulgarian food supplements are presented which contain compounds characteristic for the national traditions in nutrition, the so called Balkan diet.

**Methods:** The first one had a beneficial effect on the functions of the gastro-intestinal tract. This is a plant-derived product containing extracts from Golden shower, Cumin, Chebulic myrobalan, Wild grape, Cinnamon, Bay leaft, Cardamom, Ginger, Black pepper, Long pepper, Coriander and Asafoetida. The compounds are rich of etheric oils and resins. It is established that this food supplement has spasmolytic, anti-pain, carminative, antiemetic and antioxidant effects. The second one contains resveratrol which is a polyphenolic compound naturally found in grape, red wine, grape juice, peanuts.

**Results:** Resveratrol is shown to have a profound antioxidant effects as part of the traditional Bulgarian food products.

**Conclusions:** Our study suggests that resveratrol could improve specific metabolic parameters in individuals at risk for type

2 diabetes as well as in obese subjects with impaired glucose tolerance.

**Keywords:** Food supplements, Balkan diet, Resveratrol, Antioxidant effect, Obesity.

**Background and objectives:** Activation of basophilic cells and mast cells trigger allergic and inflammatory responses through the release of mediators such as histamine and arachidonic acid metabolites, and modulates immune responses through the production of cytokines and chemokines. Recently, allergic patients are increasing all over the world. Thus, food ingredients that suppress and alleviate allergic and inflammatory responses are required. In this study, we, therefore, investigated the effect of enzymatically-synthesized glycogen (ESG) on the degranulation and production of inflammatory cytokines and arachidonic acid metabolites in RBL-2H3 cells and clarified its underlying mechanism.

**Methods:** Caco-2 cells were differentiated as intestine monolayer to incubate for 2 weeks, bone marrow-derived mast cells (BMMC) were prepared from 10 weeks old mice and incubated in medium supplemented with interleukin (IL)-3 for 3 weeks. Allergic and inflammatory response was performed using not only RBL-2H3 cells and BMMC but also co-culture of Caco-2 cells and RBL-2H3 cells using a transwell system. Degranulation of RBL-2H3 cells or BMMC was monitored by measuring β-hexosaminidase activity. The β-hexosaminidase released into the medium or remaining within the cells (Triton X-100 extract) were determined using a colorimetric assay with p-nitrophenyl-N-acetyl-β-glucosaminide and expressed as the percentage of total activity. Production and release of tumor necrosis factor-α (TNF-α), IL-6 and arachidonic acid metabolites in/from cells were determined using western blotting and enzyme-linked immunosorbent assay system. Phosphorylation of intracellular molecules was detected using western blotting.

**Results:** ESG inhibited release of β-hexosaminidase from RBL-2H3 cells and BMMC. In Caco-2/antigen-stimulated RBL-2H3 cells, ESG inhibited release of β-hexosaminidase and production of TNF-α, IL-6, and arachidonic acid metabolites. Moreover, ESG inhibited phosphorylation of Syk, Lyn, and PLCγ1/2 as the intracellular mediators for degranulation, that of Akt, p38, and JNK as the mediators for production of TNF-α and IL-6, and that of ERK1/2 and cPLA2 as the mediator for production of arachidonic acid metabolites.
# EFFECTS OF BRAN PARTICLE SIZE ON GLYCEMIC RESPONSES. A RANDOMIZED CLINICAL TRIAL IN HEALTHY HUMANS

(1)Emilia Aimilia Papakonstantinou; (2)Mandala, Ioanna; (3)Papalexi, Katerina; (4)Chaloulos, Panagiotis.

(1)PhD. Lecturer in Nutrition and Metabolism. Department of Food Science and Human Nutrition. Agricultural University of Athens. Greece; (2)PhD. Associate Professor. Department of Food Science and Human Nutrition. Agricultural University of Athens. Greece.; (3)MS. Nutritionist. Department of Food Science and Human Nutrition. Agricultural University of Athens. Greece.; (4)BS. Nutritionist. Department of Food Science and Human Nutrition. Agricultural University of Athens. Greece.

**Background and objectives:** Diets characterized by low glycemic index (GI) and/or glycemic load (GL) have been associated with decreased risk of chronic diseases. Breads' grain particle size may affect glycemic response, possibly due to the altered starch surface, hence its susceptibility to enzymatic hydrolysis. However, the role of bran's particle size alone has not been studied. The aims of this study were a) to determine the GI and GL of three breads and b) to investigate the effects of bran's particle size on postprandial glycemic response.

**Methods:** Ten healthy, normal-weight subjects participated in GI investigation following International Standard Organization's protocol. Participants, in this double blind, crossover trial, consumed in random order 50g available carbohydrate portions as glucose (Glu, reference food, tested three-times) and white bread (WB), bread enriched with coarse wheat bran (CB), bread enriched with fine wheat bran (obtained by milling of the coarse bran) (FB) and the FB in which 10% of the flour was substituted with carob seed flour (CSFB), tested once, on separate occasions. Capillary blood glucose and salivary insulin samples were taken at 0,15,30,45,60,90 and 120min.

**Results:** CB was a high GI (70), low GL (10) bread; FB was a high GI (82), medium GL (12) bread; and CSFB was a high GI (70), low GL (8) bread. There was a significant glucoseXtime interaction (p<0.001), a glucoseXbread interaction (p=0.03) and a glucoseXtimeXbread interaction (p<0.001). There was a significant main effect of bread type on iAUC for glucose (p=0.01). Compared to the reference food, blood glucose concentrations were significantly lower: for CB at 120min (p=0.02); for FB at 15, 30, and 120min (p=0.049, p=0.004 and p=0.008, respectively); for CSFB at 15 and 30min and marginally at 45min (p=0.0008, p=0.008, p=0.06, respectively). Peak glucose rise was significantly lower only for CSFB compared to the reference food (p=0.03). No differences were observed for fasting glucose, fasting salivary insulin, iAUC for salivary insulin, time to peak rise for glucose or insulin.

**Conclusions:** Breads differed in GI/GL. Bran particle size and flour substitution by carob seed flour attenuated postprandial glycemic response, which may offer advantages to glycemic control.

**Keywords:** Bran, Particle size, Glycemic response, Carob.

# NIGAICHIGOSIDE F1 FROM LANBUZHENG (GEUM JAPONICUM THUNB.) INHIBITS OLEIC ACID-INDUCED HEPATIC STEATOSIS IN HEPG2 CELLS

(1)Cheng, Xiangrong; (2)Xia, Shufang; (3)Sun, Jing; (3)Tang, Xue; (3)Le, Guowei; (3)Shi, Yonghui.

(1)Associate professor. School of Food Science and Technology. Jiangnan University. People's Republic of China; (2)Wuxi School of Medicine. Jiangnan University. People's Republic of China;

(3)School of Food Science and Technology. Jiangnan University. People's Republic of China.

**Background and objectives:** Lanbuzheng (Geum japonicum Thunb.) is a wild vegetable in Guizhou province, China, which has also been used as traditional herbs as diuretics and astringents. Nigaichigoside F1 (NF1), the major triterpenoids in Lanbuzheng, has been report for its potent hypolipidemic and hypoglycemic activities. Thus, we hypothesized that NF1 would attenuate events leading to non-alcoholic fatty liver disease (NAFLD).

**Methods:** The culture media of HepG2 cells were added 1.0 mM oleic acid to induce hepatic steatosis. After 24 h, significant accumulation of lipid droplets were observed by Oil-Red-O based colorimetric assay, together with increased triacylglycerol (TAG) and malondialdehyde (MDA).

**Results:** The oxidative stress of HepG2 cells was significantly increased, which characterized by the content of reactive oxygen species (ROS), total antioxidant capacity (T-AOC), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (Gpx), and the ratio of reduced and oxidized glutathione (GSH/GSSG). NF1 was purified from Lanbuzheng extract by Diaion HP-20 resin (60% ethanol elution, 3 BV/h) and characterized by ESI-MS, 1H-NMR, and 13C NMR. NF1 (10 uM) increased cell proliferation by 2.35 folds with decreased TG (43%), MDA (62%), and ROS (78%) content. The content of T-AOC, SOD, CAT, Gpx, and the ratio of GSH/GSSG were increased by 2.3, 1.27, 2.15, 1.05, and 1.72 folds, respectively. Moreover, RT-PCR results confirmed NF1 (10 uM) inhibited SREBP-1c gene expression while promoted HSL and Nrf2 gene expression.

**Conclusions:** In conclusion, NF1 effectively ameliorated NAFLD symptoms by decreased TAG accumulation and increased antioxidant capacity in oleic acid-induced hepatic steatosis in HepG2 cells.

**Keywords:** Geum japonicum Thunb., Nigaichigoside F1, Non-alcoholic fatty liver disease, Antioxidant.
THE TIME COURSE OF ACUTE EFFECTS OF THREE
DOSES OF CAFFEINE ON ATTENTION, MOTOR
SPEED AND MOOD

**Background and objectives:** Moderate doses of 75–200mg caffeine have well established effects on measures of alertness and reaction time. However, less is known of the impact of lower doses that are more reflective of levels found in a cup of tea. Similarly, studies of behavioural effects of caffeine have focused on a 30–120 minute window, despite the demonstration that salivary caffeine levels remain elevated at 5 hours post-consumption. The aim of the current study was to explore effects of 40, 80 and 120mg caffeine over an 8-hour period.

**Methods:** A randomised, placebo-controlled, double-blind, parallel groups design was employed to explore the effects of caffeine (40, 80 and 120mg) on choice reaction time (CRT), tapping, ‘alertness’ and ‘sleepiness’. Participants (n=40 per group) were healthy adult caffeine consumers (mean intake 287mg/d) required to abstain from consuming caffeine overnight prior to morning testing. Following baseline, post-drink assessments commenced at 2, 40, 100, 190, 280, 370 and 460 minutes.

**Results:** Significantly faster CRT was observed following all doses of caffeine compared to placebo. A trend towards a treatment x session interaction indicated beneficial effects of 40mg on CRT were still evident at 460 minutes post-drink, whilst effects of 80 and 120mg dissipated at 280 minutes. Increased ‘alertness’ and/or decreased ‘sleepiness’ was evident up until 190 minutes post-drink following 80 and 120mg but decreased ‘alertness’ and increased ‘sleepiness’ was observed later in the day. Conversely, 40mg decreased ‘sleepiness’ from 280-460 minutes, with no effects on ‘alertness’. Interestingly, effects on CRT and mood were observed during the 2 minutes post-drink session.

**Conclusions:** These findings demonstrate that 40mg caffeine can modulate CRT and subjective ratings of alertness and sleepiness. Effects on CRT were observed over a time period spanning from 2 minutes to 8 hours post-drink. These findings are important due to an increasing desire to limit caffeine intake. They indicate that 40mg caffeine can improve reaction time without any initial subjective ‘arrousing’ effect, which may be preferable to some consumers. Further research should explore individual differences in responses to caffeine, to ascertain factors which may impact upon its dose and time-course effects.

**Keywords:** Cognition, Alertness, Caffeine, Low-dose, Time-course.

**Conflict of Interest Disclosure:** This research was funded by GlaxoSmithKline

PROTECTIVE EFFECTS OF HYDROLYZED NUCLEOPROTEINS FROM SALMON MILT AGAINST ETHANOL-INDUCED LIVER INJURY IN RATS

**Background and objectives:** Oral administration of nucleic acids from salmon milt have physiological functions in the cellular metabolism, proliferation, differentiation and apoptosis of human small intestinal epithelial cells. In this study, we examined the effects of DNA-rich nucleic acids prepared from salmon milt (DNSM) on the development of liver fibrosis in an in vivo ethanol-carbon tetrachloride (CCl4) cirrhosis model.

**Methods:** DNSM was water-solubilized using nuclease and protease. Male Wistar rats were divided into four groups. Group 1 was the control; Group 2 was treated with ethanol and CCl4; Group 3 was treated with CCl4 alone; Group 4 was treated with ethanol, CCl4 and 0.12% DNSM. CCl4 (0.1 ml/kg of body weight diluted with olive oil to 25%) was administered by intraperitoneal injection twice a week, and 5% ethanol was administered in the drinking water ad libitum. The rats were euthanized after three weeks.

**Results:** Plasma aspartate transaminase and alanine transaminase were significantly less active in the DNSM-treated group than in the ethanol plus CCl4-treated group. Collagen accumulation in the liver and hepatic necrosis were observed histologically in ethanol plus CCl4-treated rats; however, DNSM-treatment fully protected rats against ethanol plus CCl4-induced liver fibrosis and necrosis. Furthermore, we examined whether DNSM had a preventive effect against alcohol-induced liver injury by regulating the cytochrome p450 2E1 (CYP2E1)-mediated oxidative stress pathway in an in vivo model. In this model, CYP2E1 activity in ethanol plus CCl4-treated rats increased significantly, but DNSM-treatment suppressed the enzyme's activity and reduced intracellular thiobarbituric acid reactive substances levels. Furthermore, the hepatocytes treated with 100 mM ethanol induced increase in cell death and were not restored to the control levels when treated with DNSM, suggesting that digestive products of DNSM are effective for the prevention of alcohol-induced liver injury. Deoxyadenosine was suppressed the ethanol-induced increase in cell death and increased the activity of alcohol dehydrogenase.

**Conclusions:** These results suggest that DNSM treatment represents a novel tool for the prevention of alcohol-induced liver injury.

<footnote>(1)Haskell-Ramsay, Crystal; (1)Jackson, Philippa; (2)Forster, Joanne;</footnote>

<footnote>(3)Robertson, Bernadette; (1)Kennedy, David.</footnote>

<footnote>(1)PhD. Psychologist. Brain, Performance and Nutrition Research Centre. Northumbria University. United Kingdom.;</footnote>

<footnote>(2)Psychologist. Brain, Performance and Nutrition Research Centre. Northumbria University. United Kingdom.; (3)PhD candidate. Psychologist. Brain, Performance and Nutrition Research Centre. Northumbria University. United Kingdom.</footnote>

<footnote>(1)Matsui-Yuasa, Isao; (2)Goto, Mayu; (2)Yoshikawa, Eri; (3)Morita, Yuri;</footnote>

<footnote>(4)Sekiguchi, Hirotaka; (4)Sutoh, Keita; (5)Usumi, Koji; (6)Kojima-Yuasa, Akiko.</footnote>

<footnote>(1)Department of Food and Human Health Sciences. Graduate School of Human Life Science. Osaka City University.; (2)M.Sc. Graduate student. Department of Food and Human Health Sciences. Graduate School of Human Life Science. Osaka City University.; (3)B.Sc. Student. Department of Food and Human Health Sciences. Graduate School of Human Life Science. Osaka City University.; (4)Ph.D. Researcher. Life Science Institute Co. Ltd.; (5)Ph.D. President. Life Science Institute Co. Ltd.; (6)Ph.D. Associate professor. Department of Food and Human Health Sciences. Graduate School of Human Life Science. Osaka City University.</footnote>
**Keywords:** DNA-rich nucleic acid prepared from salmon milt (DNSM), plasma aminotransferases (AST and ALT), liver fibrosis, CYP2E1 activity, alcohol-induced liver injury.

## EFFECTS OF 6 MONTH RESVERATROL SUPPLEMENTATION ON VERBAL MEMORY PERFORMANCE IN HEALTHY ELDERLY ADULTS - A RAN-DOMIZED CONTROLLED TRIAL

(1)Huhn, Sebastian; (2)Beyer, Frauke; (3)Zhang, Rui; (4)Stumvoll, Michael; (5)Villringer, Arno; (6)Witte, A. Veronica.

(1)MSc. Nutritional Sciences. Department of Neurology. Max Planck Institute for Human Cognitive and Brain Sciences Leipzig. Germany.; (2)MSc. Medical Radiation Sciences. Department of Neurology. Max Planck Institute for Human Cognitive and Brain Sciences Leipzig. Germany.; (3)MSc. Nutritionist. Department of Neurology. Max Planck Institute for Human Cognitive and Brain Sciences Leipzig. Germany.; (4)MD. Medicine. Faculty of Medicine. IFB Adiposity Diseases Leipzig. Germany.; (5)MD. Medicine. Department of Neurology. Max Planck Institute for Human Cognitive and Brain Sciences Leipzig. Germany.; (6)PhD. Biologist. Department of Neurology. Max Planck Institute for Human Cognitive and Brain Sciences Leipzig. Germany.

**Background and objectives:** Resveratrol (RSV) is assumed to have beneficial effects on cognitive performance and brain-structure in humans (Huhn et al.2015). RSV, a polyphenol, occurs naturally in blueberries, peanuts, red grapes and red wine (Smoliga et al.2011). Animal models suggest that RSV is antioxidative, anti-inflammatory and mimics caloric-restriction (Athar et al.2007;Dal-Pan et al.2011). These effects might contribute to the mechanisms underlying the benefits of RSV on the metabolism, cardiovascular system and nerve-tissue found in human studies (Huhn et al.2015).

More recently, research focused on the impact of RSV on cognition. In that line, this study's aim was to evaluate the effect of 26 weeks RSV supplementation on measures of the California Verbal Learning Task(CVLT).

**Methods:** Subjects: 60 healthy elderly subjects (age60-78, mean68.19±5.16SD, anthropometry: table1, Drop-outs=7, randomized to RSV (N=27; 200mg/day) or placebo-supplementation (N=26) for 26 weeks.

Exclusion: stroke, major brain-pathologies, central-nervous-medication.

Measures: Blood-parameters, anthropometry, neuropsychological-test-battery including California-Verbal-Learning-Task.

Statistical Analysis: repeated-measures ANOVA was performed in SPSS (IBM,Version24).

Within-Subject-Factor: time

Within-Subject-Variables: CVLT-measures(Learning-sum, forgetting-rate, delayed-recall, recognition).

Between-Subject-Factors: RSV/placebo.

Covariates: age, sex, education.

**Results:** The preliminary results of the repeated-measures ANOVA did not reveal any significant time by intervention-group interaction on measures of the CVLT (all p>0.14). Compliance according to pill-counts and pill-diaries was high. Continuing analyses incorporate blood-parameters and additional cognitive measures.

**Conclusions:** The preliminary results of our study did not indicate significant effects of 26 weeks RSV supplementation on performance in the CVLT in healthy elderly subjects. This might partly be explained by a healthy sample that performed well already at baseline and a small scope to improve cognitive performance. Additionally, duration and/or dosage of RSV might have been insufficient to induce changes.

These results are in line with previous studies that did not find effects of RSV on cognitive measures (Köbe et al.2017;Wightman et al.2015). However, Witte et al.(2014) found, besides a moderate effect on cognition, significant changes in functional connectivity of the hippocampus, which is involved in memory processes. With subsequent analysis of additional cognitive measures, blood-parameters and sensitive high-resolution magnetic-resonance-imag-es, we aim to further characterize the effect of RSV on cognition and brain structure. This will help to clarify the potential of RSV to improve or maintain cognitive functions in healthy elderly adults.

**Keywords:** Resveratrol, Cognition, Memory, Longitudinal, Elderly.

## THE ANTIOXIDANT PROPERTIES AND FATTY ACIDS PROFILE IN ORGANIC SAMPLES OF LEEKS (ALLIUM AMPELOPRASUM VAR. PORRUM) AND GARLIC (ALLIUM SATIVUM L.) IN NATURA

(1)Guizellini, Gloria Maria; (2)Rodrigues Sampaio, Geni; (3)Pereira da Silva, Cintia; (4)Manólio Soares Freitas, Rosana Aparecida; (5)Ferraz da Silva Torres, Elizabeth Aparecida.

(1)Nutrition Student. Nutrition Department. School of Public Health. University of São Paulo. USP. São Paulo. Brazil.; (2)PhD. Nutrition Department. School of Public Health. University of São Paulo. USP. São Paulo. Brazil.; (3)MSc. Nutrition Department. School of Public Health. University of São Paulo. USP. São Paulo. Brazil.

**Background and objectives:** The primary goal of organic agriculture is to produce foods safer than conventional agricultural foods that are produced using synthetic fertilizers and pesticides forbidden in organic systems. Studies have shown that growing conditions may influence the nutritional composition of different types of vegetables. Leeks and Garlic are source of various biologically active phytomolecules, including organosulfur compounds and polyphenols. They have been playing an important role in the human diet and in medicine for centuries. The aim of this study was to determine the antioxidant activity, proximate composition and fatty acids content of organics leeks (Allium ampeloprasum var. porrum) and garlic (Allium sativum L.).
**Methods:** The organics leeks (L) were analyzed with different solvents, methanol (70:30,v.v) (LM) and water (LW), garlic also (G), methanol extract (GM) and water (GW). The samples were characterized for total phenolic compounds and antioxidant activity using three different methods: DPPH, β-carotene/linoleic acid system and the test of oxygen radicals absorbance capacity (ORAC).

**Results:** Methanol:water (70:30,v.v) extract produced the best results the samples of organics leek for total phenolic compounds(1.23 ± 0.4 mg/EAG g). The water extract of garlic in natura (3.04 ± 0.7 mg/EAG g) indicates the best results of samples in the same analysis. As predominant palmitic, linoleic and linolenic acids present in leeks samples and in garlic in natura were palmitic and linoleic acid. The inhibition of β-carotene/linoleic acid peroxidation (%IOL) is better in LM and with different solvents presents similar results between them GM and GW (57.15 ± 2.2% e 57.59 ± 1.5 %). The ORAC value of LMA and LW (88.45 ± 8.6 ET µmol/g and 47.09 ± 1.2 ET µmol/g) reduce in compare the samples of GM e GW (112.52 ± 0.1 ET µmol/g e 65.68 ± 4.5 ET µmol/g). The LM, LW, GM and GW exhibited antioxidant activity by DPPH method.

**Conclusions:** These results suggested that organic leeks and garlic presented antioxidant activity and contained significantly levels of phenolics. The consumption of leeks and garlic in natura can be able to provide significant health protection in order to prevent chronic diseases.

**Keywords:** Antioxidant, Allium ampeloprasum, Allium sativum, Bioactive compounds, Fatty acids.

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EFFECTS OF DIETARY PHENOLIC COMPOUNDS
FROM SOUTH AMERICAN PLANTS ON FAT-
TY ACID-MEDIATED INTERLEUKIN-6 RELEASE
FROM MAMMALIAN GLIAL CELLS

(1)Cittadini, María Cecilia; (2)García-Estéves, Ignacio; (2)Escribano-Bailón, María Teresa; (2)Rivas-Gonzalo, Julián C.; (3)Valentich, Mirta A.; (3)Repossi, Gastón; (3)Soria, Elio A.

(1)Nutricionist. Instituto de Investigaciones en Ciencias de la Salud (INICSA). Facultad de Ciencias Médicas (Universidad Nacional de Córdoba) y CONICET. Córdoba. Argentina. Instituto Nacional del Cáncer. Ministerio de Salud de la Nación. Argentina.; (2)PhD. Grupo de Investigación en Polifenoles. Facultad de Farmacia. Universidad de Salamanca. Salamanca. Spain.; (3)PhD. Instituto de Investigaciones en Ciencias de la Salud (INICSA). Facultad de Ciencias Médicas (Universidad Nacional de Córdoba). CONICET. Córdoba. Argentina.

**Background and objectives:** Phenolic compounds are secondary metabolites of plants, which can exert beneficial effects on the central nervous system after food intake. Thus, our aim was to identify anti-inflammatory phenolic compounds from South American plants on mammalian glial cells, which play a central role in the neuro-immune regulation.

**Methods:** Phenolic compounds were extracted from Lantana grisebachii (LG), Aspidosperma quebracho-blanco (AQB), and Ilex paraguariensis (IP) teas and identified by HPLC-DAD-MS. Different extract concentrations (0-200 µg/mL) were tested in human T98-G and rat C6 cell lines of glioma. Cellular viability (by the resazurin assay), fatty acid profile (by gas chromatography) and pro-inflammatory interleukin-6 release (IL-6 by ELISA) were determined. Data were analysed by partial least square regression, to discriminate bioactive phenolic compounds as predictor variables and to correlate cellular responses.

**Results:** Twenty-one compounds, which were mainly iridoids, were determined in LG. Their geniposidic derivatives (3.3%) were significantly linked to lipid polyunsaturation, but not to IL-6 release. Thirty-one compounds were found in AQB, mostly hydroxybenzoic derivatives, of which only isorhamnetin-3-O-rutinoside (0.9%) was positively related to IL-6 release. Twenty-three compounds were identified in IP, including several caffeoylquinic derivatives. Chlorogenic (24.4%) and 1-caffeoylquinic (15.0%) acids modified glial lipids increasing ω-7 palmitoleic fatty acid, which was related to IL-6 decrease.

**Conclusions:** This work enhances phytochemical knowledge of three widely distributed plants, which have not been thoroughly studied as sources of bioactive phenols. Our results suggest that the 1-caffeoylquinic and chlorogenic acids provided by I. paraguariensis tea can exert anti-inflammatory effects, which would be significant for human nutrition and health given its high intake by South American populations.

**Keywords:** Chlorogenic acid. 1-caffeoylquinic acid. Iridoid. Isorhamnetin. Neuroinflammation. Ilex paraguariensis.

**Conflict of Interest Disclosure:** We confirm that there are no known conflicts of interest associated with this presentation and there has been no significant financial support for this work that could have influenced its outcome.

144/1697

QUANTIFICATION OF BIOACTIVE COMPOUNDS
IN CUPUAÇU ALMONDS (THEOBROMA GRAN-
DIFLORUM)

(1)Pontes de Andrade, Fátima Eliana; (2)Kwiatkowski, Angela; (3)da Silva Coimbra, Paula; (4)dos Santos Souza, Giovani; (3)Dias Pereira, Queila; (3)Marçal Chaves, Roseni; (3)de Oliveira Costa, Camila Cristina; (5)Bergamo, Joseila Aparecida.

(1)Graduate Student Food Technology. Federal Institute of Mato Grosso do Sul. Brazil.; (2)Post doctor in Food Science. Food Technology Coordination. Federal Institute of Mato Grosso do Sul. Coxim. Brazil.; (3)Graduate Student in Food Technology. Federal Institute of Mato Grosso do Sul. Coxim. Brazil.;
(4)Graduate Student in Chemistry. Federal Institute of Mato Grosso do Sul. Coxim. Brazil.; (5)Biologist. Chemistry Laboratory. Federal Institute of Mato Grosso do Sul. Coxim. Brazil.

**Background and objectives:** The fruit of cupuaçu is used in Brazilian cuisine to prepare juices, ice creams, liquors, creams,
puddings, cakes, sweets and sweets, due to the aroma and the ac-
centuated acidity, much appreciated by consumers. Through the
feeding we obtain macro and micronutrients and also bioactive
compounds, health promoters, as the antioxidant, anti-inflam-
matory and hypocholesterolemic action of some of these com-
pounds. The antioxidant activity may be related to the presence
of vitamins and phenolic compounds. The objectives of this work
were to characterize cupuaçu almonds in relation to pH and total
acidity and to quantify bioactive compounds, such as vitamin C
determination, phenolic compounds and antioxidant activity for
the preparation of food products.

**Methods:** The cupuaçu almonds were separated from the fruit
pulp and dried in an oven with air circulation at 60°C for 48 hours.
The pH values were determined by digital potentiometer, titratable
total acidity, vitamin C by the titration method according to the
Adolfo Lutz Institute's analytical standards (2005), phenolic com-
pounds by the Follin Ciocauteau method (Bucic-Kojic et al., 2007)
Of the curve performed with gallic acid (200, 400, 600, 1000 and
1400 mg/L), and antioxidant activities by the capture of free radi-
cal DPPH (2,2-Diphenyl-1-picryl-hydrazyl) (Mensor et al., 2001 ).

**Results:** The pH value of the dehydrated almond was 4.44. The total acidity resulted in 0.029 g/100g. The vitamin C content was 272.73 mg/100g. The content of phenolic compounds obtained was 364.50 mg equivalent of gallic acid/100 g, 60% higher than that reported in the literature for fresh almonds (Pugliese, 2010), difference due to the process of dehydration concentrate the nutrients. The antioxidant activity resulted in 95.86%. The results show that the cupuaçu almond presents high content of vitamin C, phenolic compounds with significant antioxidant activity.

**Conclusions:** From the cupuaçu almond can be elaborated several food products with high nutrient content with a health action of vitamin C and phenolic compounds, such as antioxidant activity.

**Keywords:** Theobroma grandiflorum, Almonds, Vitamin C, Phenolic compounds, Antioxidant activity.

blood glucose levels by increasing muscle glucose uptake in an in-
sulin-independent manner. Insulin stimulates glucose uptake in
muscle via translocation of glucose transporter 4 (GLUT4) vesicles
to the plasma membrane. GLUT4 translocation involves the PI3K/
Akt pathway. There are a few reports that show that isoleucine pro-
motes GLUT4 translocation, but the details of the mechanism by
which isoleucine increases glucose uptake in skeletal muscle are
not yet fully understood. The purpose of this study was to clarify
the mechanism of glucose uptake into skeletal muscle by isoleu-
cine.

**Methods:** Male Wistar rats were food deprived for 18 h and then administered 1.35 g L-isoleucine/kg body weight (prepared as 54.0 g/L of the L-amino acid in distilled water) by oral gavage. GLUT4 translocation in skeletal muscle (gastrocnemius, soleus and EDL) was detected by immunofluorescent staining. We also evaluated the phosphorylation status of Akt and AMPK involved in glucose uptake evoked by insulin using C2C12 myotubes treated with isoleucine. Prior to the experiments, differentiated C2C12 myotubes were starved for 3 h in a Krebs-Henseleit-Hepes buffer and then treated with 1-10mM isoleucine for 30 min.

**Results:** The administration of isoleucine stimulated translocation of GLUT4 vesicles to the plasma membrane in all examined skeletal muscles. At the site of the plasma membrane, GLUT4 was more abundantly expressed in EDL of isoleucine administered rats as compared to other muscles. The phosphorylation of Akt was increased by isoleucine in C2C12 myotubes. On the other hand, there was no change in phosphorylation of AMPK.

**Conclusions:** EDL (fast twitch muscle) is more sensitive to the effect of isoleucine as compared to other muscles. Isoleucine seems to stimulate translocation of GLUT4 in skeletal muscle through a mechanism distinct from that of insulin.

**Keywords:** Isoleucine, Skeletal muscle, Glucose uptake, GLUT4, PI3K.

144/1715

ANALYSIS OF THE MECHANISM OF ISOLEUCINE INDUCED GLUCOSE UPTAKE INTO SKELETAL MUSCLE

(1)Shiga, Kana; (2)Takami, Yuki; (3)Narita, Kazusa; (4)Yoshizawa, Fumiaki; (5)Sato, Yusuke.

(1)MSc. Student. Department of Agrobiology and Bioresources. Faculty of Agriculture. Utsunomiya University. Japan.; (2)MSc. Student. Department of Agrobiology and Bioresources. Faculty of Agriculture. Utsunomiya University. Japan.; (3)MSc. Department of Agrobiology and Bioresources. Faculty of Agriculture. Utsunomiya University. Japan.; (4)PhD.Professor.Utsunomiya University. Japan.; (5)PhD. Associate Professor. Utsunomiya University. Japan.

**Background and objectives:** Branched-chain amino acids (leucine, isoleucine, and valine) are essential amino acids and necessary for proper muscle function. Isoleucine is known to lower

144/1734

A POLYPHENOL-RICH CALAFATE EXTRACT RE-
VERTS INSULIN RESISTANCE IN MICE FED WITH
CAFETERIA DIET

(1)Garcia-Diaz, Diego; (2)Soto-Covasich, Jessica; (3)Reyes-Farias, Marjorie; (4)Jimenez, Paula; (5)Almeyda, Rocio; (5)Cortez, Andrea;

(5)Gonzalez, Francisca; (5)Malebran, Paula; (5)Marambio, Macarena.

(1)PhD. Department of Nutrition. School of Medicine. University of Chile.; (2)Biochemist. Instituto de Biologia. Facultad de Ciencias. Pontificia Universidad Catolica de Valparaiso. Chile. Departamento de Nutricion. Facultad de Medicina. Universidad de Chile. Chile.; (3)Biochemist. Departamento de Nutricion. Facultad de Medicina. Universidad de Chile. Chile.; (4)Veterinary. Departamento de Nutricion. Facultad de Medicina. Universidad de Chile. Chile.; (5)Nutricionist. Departamento de Nutricion. Facultad de Medicina. Universidad de Chile. Chile.

**Background and objectives:** Insulin resistance is a world-
wide high prevalent disease accompanied usually by a chronic
low-degree inflammatory state. Calafate is a Chilean native berry with a high polyphenol content. Polyphenols have been shown to possess anti-inflammatory properties. We aim to determine the effect of a Calafate extract on the development of insulin resistance in mice fed on cafeteria diet.

**Methods:** By using of an Amberlite XAD7HP column a purified and polyphenol-rich calafate extract was prepared. Total polyphenols, total anthocyanins and antioxidant activity were determined. Male C57BL/6 mice were fed on cafeteria or standard diet for 14 weeks. From week 10th, mice received the calafate extract in water (daily doses of 50mg [total polyphenols] /Kg). Body weight, food and water intake were registered. IPGTT were performed at 9 and 13 weeks and p-AKT was evaluated to determine insulin signaling pathway activity. Also, insulin concentration in serum samples was measured.

**Results:** The purified calafate extract presented a total polyphenol content, anthocyanin content and antioxidant capacity of 2242.3 mg GAE/100g DW, 66.1 mg C-3-G/100g DW and 38.43 mmol Fe+2/100g DW, respectively. No differences were observed in the insulin concentration between groups. The IPGTT performed at week 9th showed that mice fed on cafeteria diet developed insulin resistance. Interestingly, calafate extract improved the glucose tolerance at week 13th. Western blotting revealed that calafate extract increased p-AKT content, suggesting enhanced insulin signaling.

**Conclusions:** Our results show for the first time a beneficial effect of a calafate extract over insulin resistance in an in vivo model.

**Keywords:** Calafate. Polyphenols. Diabetes.

**Further collaborators:**

Financial Support: PhD Grant 21120219 (CONICYT, Chile) / PUCV Thesis Support Grant / Fondecyt 11110219 (CONICYT, Chile)

EFFECT OF THE CONSUMPTION OF HAMBURGERS PREPARED WITH ADDITION OF WINE GRAPE POMACE FLOUR, RICH IN FIBER AND ANTIOXIDANTS, ON OXIDATIVE DAMAGE MARKERS

(1)Urquiaga, Ines; (2)Perez, Druso; (3)Dicenta, Sara; (4)de la Cerda, Paula; (5)Urzua, Catalina; (6)Troncoso, Danitza; (6)Mackenna, Maria Jose; (7)ECHEVERRÍA, GUADALUPE; (8)Rigotti, Attilio.

(1)PhD. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile.; (2)MSc. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile.; (3)MSc. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile.; (4)Biochemist. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile.; (5)Food Engineering. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile.; (6)MD. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile; (7)MSc. Centro de Nutricion Molecular y Enfermedades Crónicas. Pontificia Universidad Católica de Chile; (8)PhD. Centro de Nutricion Molecular y Enfermedades Crónicas. Departamento de Nutrición, Diabetes y Metabolismo, Escuela de Medicina. Pontificia Universidad Católica de Chile.

**Background and objectives:** Healthy eating is one of the pillars of a healthy lifestyle, leading to the prevention of chronic non-communicable diseases. Antioxidants and fiber are two bio-active components with proven beneficial health properties. Currently, in Chile and the world, fiber and antioxidants intake is less than that recommended by the WHO.

Our objective was to develop a functional food, enriching a high consumption meat-based food with wine grape pomace flour (WGPF), rich in fiber and antioxidants, and to study the effect of its consumption on oxidative damage markers in an intervention study.

**Methods:** A beef hamburger (100 g) was formulated with 7% of WGPF, containing 3.5% of fiber and 1.2 mg EAG/g of polyphenols (WGPF-hamburger). WGPF is a byproduct of vinification (Cabernet sauvignon). A 3-month controlled intervention study was completed in 27 male volunteers with at least one component of the metabolic syndrome. Volunteers daily consumed one WGPF-hamburger during the first month, interrupted by a month washout period, and followed by a control month period where they daily consumed a hamburger without WGPF (control-hamburger). At the beginning and at the end of each intervention period we evaluated oxidative damage markers: Advanced Oxidation Protein Products (AOPP), carbonyls in proteins, oxidized LDL y malondialdehyde (MDA).

**Results:** Intake of WGPF-hamburger significantly decreased AOPP (p=0.005) and oxidized LDL (p=0.048) compared with control-hamburger consumption. No significant effect was observed in carbonyl groups in proteins and MDA levels between WGPF-hamburger and control-hamburger periods.
**Conclusions:** The consumption of a beef hamburger elaborated with 7% WGPF protected from oxidative damage when comparing to control-hamburger, showing its potential to prevent chronic diseases, and its functional properties. Supported by FONDEF (grant #IT14I10011) and Agrosuper.

**Keywords:** Oxidative damage, Fiber, Functional food.

## 144/1900

### ANTI-OBESITY EFFECTS OF KAEMPFEROL BY INHIBITING EARLY ADIPOGENESIS IN 3T3-L1 CELLS

(1)Torres Villarreal, Denys Armandina; (2)Ortíz López, Rocío; (3)De La Garza Hernández, Ana Laura.

(1)Bs. Nutrition. Autonomous University of Nuevo Leon, Center for Research and Development in Health Sciences, Faculty of Public Health and Nutrition; (2)School of Medicine, Tecnológico de Monterrey, México; (3)Autonomous University of Nuevo Leon. University of Public Health and Nutrition.

**Background and objectives:** Obesity is a chronic metabolic disorder defined as an abnormal or excessive fat accumulation with a health risk. Several scientific investigations are focused on search and identify different bioactive compounds that may be used as a treatment for obesity. Kaempferol is a natural flavonoid, found in fruits, vegetables and teas. Recent studies have suggested that it has anti-oxidant, anti-inflammatory and anti-obesity effects. In this context, the present study aimed to assess the anti-obesity properties of kaempferol on the anti-adipogenic and lipolytic effects and the expression of genes involved in the metabolic pathways.

**Methods:** 3T3-L1 cells were cultured in DMEM supplemented with 10% FBS and 1% antibiotics (initiation medium). At 24 hours, the differentiation was induced (day 0) with DMEM + 1 µL/mL dexamethasone, IBMX and insulin (differentiation medium (DM)), and 2 days after, the medium was replaced for fresh medium containing DMEM + 1 µL/mL insulin (post-DM). This medium was switched every 2 days. Preadipocytes (day 0) and mature adipocytes (day 12) were treated with 60 µM of kaempferol until the finishing of the treatment (day 21). At this point, cells were stained with Oil Red O to measure the lipid accumulation. Furthermore, the expression of genes involved in the adipogenesis, lipolysis and inflammation processes was evaluated with RT-qPCR.

**Results:** Kaempferol has a potential effect on adipogenesis by reducing 62% of lipid accumulation in relation to the positive control (p < 0.001). Moreover, the intracellular lipid content were diminished (39%) after treatment with kaempferol in mature adipocytes (p = 0.256). Regarding the molecular mechanisms, the treatment with kaempferol downregulated the mRNA expression of C/EBPα (p < 0.05) in comparison with the control cells. On the other hand, the expression levels of PNPLA2 (p < 0.01) and LIPE (p < 0.05), which are associated with hydrolysis of tri- and diglycerides, significantly increased in the presence of kaempferol.

**Conclusions:** These data indicate that kaempferol has anti-adipogenic effects in 3T3-L1 cells by suppressing the mRNA expression of C/EBPα and stimulating the expression of genes involved in lipid metabolism.

**Keywords:** Kaempferol, Obesity, Adipogenesis, Lipolysis, Nutrigenomic.

**Further collaborators:**

MC. Maribel Montufar. Universidad Autónoma de Nuevo León, Centro de Investigación y Desarrollo en Ciencias de la Salud.

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### EFFECT OF THE LYOPHILIZATION PROCESS ON THE CONTENT OF BIOACTIVE COMPOUNDS FROM FRUIT PULPS OF BRAZILIAN PANTANAL

(1)Kwiatkowski, Angela; (2)de Oliveira Costa, Camila Cristina; (3)Santos de Minas, Ramon; (4)dos Santos Souza, Giovani; (2)Pontes de Andrade, Fátima Eliana; (2)Marçal Chaves, Roseni.

(1)Dra. Agronomy. Food Technologist. Food Technology Coordination. Federal Institute of Mato Grosso do Sul. Coxim. Brazil.; (2)Graduate student of Food Technology. Federal Institute of Mato Grosso do Sul. Coxim, Brazil.; (3)Dr. Vegetable Production. Professor. Federal Institute of Mato Grosso do Sul. Coxim, Brazil.; (4)Graduate student of Chemistry. Federal Institute of Mato Grosso do Sul. Coxim, Brazil.

**Background and objectives:** The Pantanal is a biome situated in the state of Mato Grosso and Mato Grosso do Sul, Brazil. The vegetation of the Pantanal presents diverse diversity due to the influences provinces located to its surroundings, like Forest Amazon, Cerrado and others. Fruits are plentiful, but recently they are in focus by the research community. Thus, cupuaçu (Theobroma grandiflorum), noni (Morinda citrifolia) and cajá-manga (Spondias dulcis) can be classified as food with potential use in nutrition, providing several nutrients and bioactive compounds that can Assist in the prevention of degenerative diseases. The drying process by freeze drying helps to increase shelf life. The objective of this work was to evaluate and compare the levels of organic acids present and bioactive compounds, antioxidant capacity of fruit pulps of cupuaçu, noni and cajá-manga.

**Methods:** The pulps of the fruits of cupuaçu, noni and cajá-manga were analyzed. The pulps were lyophilized the pulps were frozen, at -40 °C for 24 hours in ultrafreezer, and left under lyophilization for 48 hours in the absence of light. The values of pH, total acidity, determination of phenolic compounds by the Follin-Ciocauteau method, total flavonoids and antioxidant activities were determined by the DPPH (2,2-Diphenyl-1-picryl-hydrazyl) free radical.

**Results:** For cupuaçu pulp the results were: pH of 3.26; Acidity of 0.182 g / 100g; Content of phenolic compounds obtained was 130.80 mg equivalent of gallic acid / 100 g; Flavonoid content was 14.06 mg / 100g, with antioxidant activity 96.50%. Noni: pH 3.95; Acidity of 0.119 g / 100g; Phenolic compounds was 43.30