gy x-ray absorptiometry scanning (DXA) and was also estimated using the predictive equations: Hume I and Hume II; Salamat et al. and Kulkarni et al. I and Kulkarni II. ANOVA one-way and Tukey's post-hoc test was performed to compare the values of LM measured by DXA and estimated by equations. Pearson's correlation was performed to verify the correlation of LM quantified by equations and DXA. Bland-Altman analysis evaluated the percentage of underestimation and overestimation of the equations.

**Results:** It was observed that Salamat et al and Kulkarni II equations resulted in the same values of LM as measured by DXA (36.62±4.47; 38.86±5.67 and 36.59±4.56, respectively). The equations of Hume I and Kulkarni I estimated higher values (41.71±5.23 and 46.29±6.47, respectively), while Hume II lower values (19.72±4.00) of LM than that quantified by DXA. Hume I, Salamat et al., Kulkarni I, Kulkarni II and Hume II equations showed a positive correlation with LM values of DXA (r=0.89; r=0.69; r=0.87; r=0.89, and r=0.81, respectively). In the Bland-Altman analysis it was observed that Salamat and Kulkarni II equations overestimated in the LM 0.12% and 5.75%, respectively. Additionally, it was observed that the Hume I and Kulkarni I overestimated the LM in 13.07% and 23.22%, respectively and Hume II equation underestimated in 60.77%.

**Conclusions:** The equations that presented better prediction of LM were Salamat and Kulkarni II, but it should be used in clinical practice with caution.

**Keywords:** Lean Mass, Predictive equations, DXA

144/2815

MUSCLE WASTING AND NUTRIENT INTAKE ASSESSMENT OF PATIENTS HOSPITALIZED WITH PRESSURE INJURY UNDER ORAL AND ENTERAL NUTRITIONAL THERAPY

(1)Diniz Camargo, Marcela Aparecida; (1)Mourão, Letícia; (2)Gonçalves Camargo, Isabela; (3)de Lucena Ferretti, Roberta.

(1)Nutritionist. University of Taubate. University Hospital of Taubate. Sao Paulo. Brazil; (2)Nutritionist. University of Taubate. University Hospital of Taubate. Sao Paulo. Brazil.; (3)PhD Student. MSc. Nutritionist. Federal University of Sao Paulo. University of Taubate. Sao Paulo. Brazil.

**Background and objectives:** Inadequate nutrients intake for hospitalized patients is of paramount importance and it is related to complications in the clinical setting, higher mortality rate, higher costs and length of hospitalization, and the occurrence of pressure injury (PI). The main objective of this study was to evaluate the muscle wasting and nutrient intake of adult and elderly patients hospitalized with PI under oral and enteral nutritional therapy.

**Methods:** A cross-sectional study was carried out with adult and elderly patients above 20 years old in a hospital in Taubate, Sao Paulo, with PI under oral or enteral nutrition therapy between February and June 2016. To assessment of muscle wasting, Calf Circumference measure was evaluated, considering cutoff point

below 31 cm to this outcome. To evaluate the nutrient intake were considered type of nutritional therapy used. For patients under Oral Nutrition Therapy, percentage of food acceptance and a food composition table (TACO) were used to calculate nutrition intake while for patients under Enteral Nutritional Therapy, a total amount of diet infused, according to the formula used, were considered. Regarding evaluation of nutrients intake (vitamin A, E, C, selenium, zinc) was considered as reference the Estimated Average Requirement (EAR) proposed by the Dietary Reference Intakes (DRIS) developed by the Foods and Nutrition Board/Institute of Medicine of the United States. Descriptive statistics were used to analyze the data.

**Results:** Among the 29 patients evaluated, 72.41% were male. It was verified that 58.6% of the patients presented muscle wasting. In relation to nutrients intake, 55.17% presented vitamin C intake above, 41.37% below and 3.44% within EAR values; 86.20% presented vitamin E intake above, 10.34% below, and 3.44% within EAR; Regarding to vitamin A, 68.96% presented above, 33.42% below and 3.44% within EAR; 55.17% presented selenium intake above EAR, 31.03% below and 13.79% within EAR; 82.75% of the patients had zinc intake above, and 17.24% below EAR.

**Conclusions:** It was concluded that the majority of patients presented muscle wasting, and there was predominance of patients with higher intake than EAR for vitamins C, E, A, zinc and selenium.

**Keywords:** Pressure Injury. Muscle Wasting. Nutrient Intake. Patients Hospitalized.

**Further collaborators:**

Marcela Aparecida Diniz Camargo, Letícia Mourão, Isabela Gonçalves Camargo helped to implement the study and data collection. Roberta de Lucena Ferretti conceived, designed, and implemented the study, collected and interpreted data, wrote and revised the text was responsible for coordinating original study and contributed to the intellectual content. All authors have sufficiently contributed to the development of this study.

144/2830

PREVALENCE OF MALNUTRITION IN HOSPITALIZED PATIENTS WITH PRESSURE INJURY

(1)Mourão, Letícia; (1)Camargo Gonçalves, Isabela; (2)Diniz Camargo, Marcela Aparecida; (2)de Souza da Cruz, Francine; (2)Moura dos Santos Botan, Gabriela; (3)de Lucena Ferretti, Roberta.

(1)Nutritionist. University of Taubate. University Hospital of Taubate. Sao Paulo. Brazil; (2)Nutritionist. University of Taubate. University Hospital of Taubate. Sao Paulo. Brazil.; (3)PhD Student. MSc. Nutritionist. Federal University of Sao Paulo. University of Taubate. Sao Paulo. Brazil.

**Background and objectives:** The hospitalized patients, especially malnourished, are likely to develop Pressure Injury (PI). Early identification of risk situations in nutritional diagnosis allows adequate planning of actions for prevention and treatment of diseases. The main objective of this study was to identify the prevalence of malnutrition in adult and elderly hospitalized patients with PI.
**Methods:** A cross-sectional study that evaluated adult and elderly above 20 years old in a hospital in Taubate, Sao Paulo, between February and June 2016. To nutritional status assessment, the measure of Calf Circumference was obtained (cutoff value below 31 cm to identify malnutrition). The Braden scale was used to identify the risk, hight and moderate, to development of another PI. C Reactive Protein (CRP) values were obtained from patient's chart to assess inflammation status (reference value above or equal 0.5 mg/dL to identify inflammation). Statistical Descriptives were used to analyse the data.

**Results:** Among the 40 patients evaluated, 62.5% were male. The mean age were 65.8 ± 16.12 and 68.0 ± 15.88 years, for female and male sex, respectively. Regarding nutritional status, 62.5% of the patients were malnourished. It was verified that 87.5% had one PI and 12.5% developed two PI, and the highest percentage of lesions found in the sacral region (69%). In relation to the risk assessment for the development of PI, 55% of patients were at high risk and the remainder 45% presented moderate risk. Considering the patients classified as having moderate and high risk of another PI development, 61.11% and 63.63% were malnourished, respectively. Regarding CRP levels, 97.5% of the patients presented high values.

**Conclusions:** It is concluded that 65.5% of the patients presented malnutrition, being propitious to the development of new PI and worse clinical outcomes.

**Keywords:** Pressure Injury. Malnutrition. Calf Circumference. Patients Hospitalized

**Further collaborators:** Letícia Mourão, Isabela Gonçalves Camargo, Marcela Aparecida Diniz Camargo, Francine de Souza da Cruz and Gabriela Moura Santos Botan helped to implement the study and data collection. Roberta de Lucena Ferretti conceived, designed, and implemented the study, collected and interpreted data, wrote and revised the text was responsible for coordinating original study and contributed to the intellectual content. All authors have sufficiently contributed to the development of this study.

evidence on the efficacy of MNPs in improving Zn status among moderately malnourished children is scarce. the study aimed at establishing bioequivalence of MNPs to conventional fortification in improving Zn status of moderately malnourished children in Thika Informal settlements, Kenya.

**Methods:** In a cluster randomized trial, twelve villages were randomized to four study groups. Three experimental groups received different formulations of MNPs added to CSB; multiple micronutrients (MMN) containing Zn (CSB-sprinkle-MMNZn), MMN without Zn (CSB-sprinkle-MMN) and Zn singly (CSB-sprinkle-Zn). The control group received conventionally Zn-fortified CSB containing MMN. CSB was prepared centrally in the participating villages and enrolled children fed daily. Dietary intake was assessed using 24-hour recall. Enrolled children underwent physical examination and anthropometric measurements. Serum Zn levels were determined pre-post intervention. Daily consumption of CSB was monitored for a period of six months. Sample size was calculated to show bioequivalence within ±20% limit. A total of 346 children (MUAC ≥11.5 - <12.5cm) were enrolled; CSB-sprinkle-MMNZn (N=84), CSB-sprinkle-MMN (N=88), CSB-sprinkle-Zn (N=93) and control group (N=81). Analysis was by intention-to-treat.

**Results:** At baseline, 64.2% of the children were Zn deficient, Zn intake was sub-optimal for 95.7% of children. A mixed effects linear regression was used to model pre-post change in serum Zn levels, adjusting for age, sex and socio-economic status. Compared to control group, the change was significantly lower for CSB-sprinkle-MMN (β=-20.0µg/dL; 95% CI: -26.5, -13.5) and significantly higher for CSB-sprinkle-MMNZn (β=5.9 µg/dL; 95% CI: 0.5, 11.3). No significant difference was observed between CSB-sprinkle-ZN and control group (β=-2.1; 95% CI: -8.3; 4.2). Conventional fortification was not bioequivalent to MNPs in improving serum Zn levels (t-tests 95% CI: -2.2; 8.9 p>0.05).

**Conclusions:** MNPs were more effective in improving Zn status of malnourished children compared to conventional fortification and are therefore recommended in fortification programmes.

**Keywords:** Malnutrition, Zinc, Deficiency, Micronutrient Powders

## BIOEQUIVALENCE OF MICRONUTRIENT POWDERS TO CONVENTIONAL FORTIFICATION ON SERUM ZINC LEVELS OF MODERATELY MALNOURISHED CHILDREN IN THIKA INFORMAL SETTLEMENTS, KENYA

**Background and objectives:** Corn Soy Blend (CSB) is the standard protocol of treating malnourished children. CSB however, has poor micronutrient bioavailability and may result into poor outcomes during nutrition recovery of the children. Micronutrient powders (MNPs) have a potential to improve micronutrient nutrition. Zinc(Zn) is an essential micronutrient for immune-protection, growth and sensory development. Scientific

## NUTRITIONAL STATUS OF ADULT AND ELDERLY HOSPITALIZED WITH SYSTEMIC ARTERIAL HYPERTENSION

**Background and objectives:** Systemic arterial hypertension (SAH) is the leading cause of cardiovascular disease in adults and the

<footnote>(1)Juliana, Kiio; (2)Sophie, Ochola; (3)Ruth, Nduati; (2)Elizabeth, Kuria.</footnote>

<footnote>(1)University of Nairobi. Kenya.; (2)PhD. Department of Food. Nutrition and Dietetics. Kenyatta University. Kenya.; (3)MBChB. MMed. MPH. Department of Paediatrics. University of Nairobi. Kenya.</footnote>

<footnote>(1)de Souza da Cruz, Francine; (1)Moura dos Santos Botan, Gabriela;</footnote>

<footnote>(2)Nunes Novais Calisto, Ariane; (2)Resende de Aguiar, Livia; (3)de Lucena Ferretti, Roberta.</footnote>

<footnote>(1)Nutritionist. University of Taubate. University Hospital of Taubate. Sao Paulo. Brazil; (2)Nutritionist. University of Taubate. University Hospital of Taubate. Sao Paulo. Brazil.; (3)PhD Student. MSc. Nutritionist. Federal University of Sao Paulo. University of Taubate. Sao Paulo. Brazil.</footnote>
elderly, according to the World Health Organization, and may worsen the clinical picture of hospitalized patients. Excess weight is strongly associated with elevated blood pressure. The main objective of this study was to evaluate the nutritional status of hypertensive adult and elderly patients admitted to a hospital in Taubate, São Paulo.

**Methods:** A cross-sectional study that evaluated adults and elderly patients diagnosed with hypertension, aged over 20 years, at a Hospital in Taubate, São Paulo, between February 2015 and May 2016. The variables evaluated were sex, age, weight and height for the calculation of body mass index (BMI). Nutritional status classification was made through World Health Organization criteria for adults, and Lipschitz criteria for the elderly, considering as parameter BMI. For statistical analysis, the t-test for independent samples and one-way ANOVA were used, with a Tukey post hoc analysis, with a significance value p <0.05.

**Results:** Among the 176 patients evaluated, 63% (n = 111) were female. The median age for the female and male sex was 65.3 ± 11.3 and 67.4 ± 13.1, respectively. There was a significant difference between the sexes (p <0.05). With data analysis, 63% (n = 71) of women were overweight, 27.9% (n = 31) presented eutrophy and the others were underweight. Regarding the males, 61% (n = 40) were overweight, 30.7% (n = 20) presented eutrophy and the others were underweight, and this difference was observed among all groups, through post hoc analysis (p <0.001).

**Conclusions:** It is concluded that there is a high prevalence of overweight in hypertensive adults and elderly, and this outcome was higher to female sex. This is considered an injury to the hypertensive patient, and intervention measures are necessary to reduce excess adiposity.

**Keywords:** Systemic Arterial Hypertension. Nutrition Status. Overweight. Body Mass Index. Hospitalized Patients.

**Further collaborators:**

Francine de Souza da Cruz, Gabriela Moura dos Santos Botan, Ariane Nunes Novais Calisto and Livia Resende de Aguiar helped to implement the study and data collection. Roberta de Lucena Ferretti conceived, designed, and implemented the study, collected and interpreted data, wrote and revised the text was responsible for coordinating original study and contributed to the intellectual content. All authors have sufficiently contributed to the development of this study.

**Background and objectives:** Patients with cancer submitted to radiotherapy (RxT) often undergo prolonged fasting, which may favor the development of malnutrition. The objectives of this study were to identify the fasting time performed by patients submitted to RxT under sedation and to verify nutritional status evolution in three moments.

**Methods:** A longitudinal study that assessed children with malignant neoplasms, aged 2 to 8 years old, at a Specialized Institute of Pediatric Oncology, Sao Paulo, from July 2015 to September 2016. Data were collected in three moments, Pre, During ans Post RxT. Weight, height/length were analyzed, to calculate the Body Mass Index (BMI). Measures of middle upper arm circumference and skinfold triciptal were obtained to calculate Arm Muscle Area (AMA). Nutritional Status was assessed through z-score of BMI, according to World Health Organization recommendation. Patients were divided into two groups, A (fasting up to 14hours) and B (above 14 hours). Δ BMI z-score were calculated (variation of BMI z-score among three moments).

**Results:** Among the 19 patients assessed, 57.8% were male. In group A (n=11), during 3 moments, there was 9%(1st),9%(2nd),9%(3rd) of leanness, respectively, 82%(1st), 82%(2nd) and 91%(3rd) of eutrophy and 9%(1st), 9%(2nd) and 0%(3rd) of obesity. Regarding to AMA percentile<5, there were 9%, 9% and 18% of patients, respectively. In group B (n=8), in 3 moments were observed 0%(1st),12.5%(2nd),12.5%(3rd) of leanness, respectively,100%(1st), 87.5%(2nd), 87.5%(3rd) of eutrophy. Regarding to AMA below adequate there were, in 3 moments,12.5%, 12.5% and 37.5%, respectively. Weight loss was present in 47.5%, 73.5% and 52.6% in three moments, respectively. Regarding to the Δ BMI z-score, there was a greater variation for worsening of the BMI z-score, for group B (Δ BMI z-score=0.8) while Group A Δ BMI z-score=0.4.

**Conclusions:** It is concluded that the mean fasting time exceeded the recommendations of ACERTO project. Nutritional Status was predominantly eutrophic with AMA within normal range, however, there was weight loss (larger decrease of the BMI z-score) in a large part of the sample, being this endpoint higher in the group of patients who underwent longer fasting.

**Keywords:** Nutritional Status. Malignant Neoplasms. Children. Radiotherapy. Fasting Time.

**Further collaborators:** Adriana Garófolo, Priscila dos Santos Maia-Lemos and Roberta de Lucena Ferretti helped to implement the study interpreted data, wrote and revised the text. Karen Jalo-

<footnote>(1) Jaloretto Teixeira Guedes, Karen; (2) de Lucena Ferretti, Roberta;</footnote>

<footnote>(3) dos Santos maia-Lemos, Priscila; (3) Garófolo, Adriana.</footnote>

<footnote>(1)Nutritionist. Federal University of Sao Paulo. Brazil; (2)PhD student. MSc. Nutritionist. Federal University of Sao Paulo. Sao Paulo. Brazil.; (3)PhD. MSc. Nutritionist. Federal University of Sao Paulo. Sao Paulo. Brazil.</footnote>
retto Teixeira Guedes conceived, designed, and implemented the study, collected and interpreted data, wrote and revised the text was responsible for coordinating original study and contributed to the intellectual content. All authors have sufficiently contributed to the development of this study.

144/2862

ASSESSMENT OF NUTRITIONAL STATUS, COM-
PARATIVE STUDY. CONCORDANCE BETWEEN
MULTIFREQUENCY, MONOFREQUENCY BIO-
ELECTRICAL IMPEDANCE ANALYSIS AND AN-
THROPOMETRY

**Background and objectives:** To determine the nutritional health of an individual, we used nutritional assessment with anthropometric techniques and bioelectrical impedance analysis (BIA), two of the field methods most used for their easy application, good reproducibility and low cost.

We aimed to establish the concordance between two cohorts
who were assessed by anthropometry and BIA.

**Methods:** We included 2871 patients between 18 and 90 years old in two cohorts of patients according to availability of bioimpedance equipment (2013-2015). Nutritional status was assessed by monofrequency BIA, multifrequency BIA and anthropometry with 4 skinfold thickness measurement. The Bland and Altman method was used to evaluate concordance between groups.

**Results:** 1168 studies were performed by BIA monofrequency, 69% (n = 807) female. Mean age 41 years (DS 15), mean BMI of 30 kg / m2 (DS 8). Mean % of body fat measured was 33.12% (DS 11), mean % of body fat measured by anthropometry was 34.3% (DS 10).

1703 multifrequency bioimpedance were performed, 70%
(1187) female. Mean age was 44 years (DS 16) and mean BMI 30.5
(DS 7.7). Mean % of body fat was 37.7 (SD 11), mean % of body fat
measured by anthropometry was 35.3 SD (12).

Bland-Altman comparison Limits of agreement ( -11.587 to 9.317)

Mean difference: -1.135 (CI -1.335 to -0.936) Range: 4.750 to 55.550

Pitman's Test of difference in variance: r =0.411, n = 2638, p =0.000

The limits of agreement were broad. At highest fat percentage averages there were difference in means between methods. This variability is not consistent across the graph.

**Conclusions:** Body composition assessment estimates by BIA monofrequency, multifrequency and skinfold method were different especially at greater body fat, concluding that these methods are not interchangeable

**Keywords:** Bioelectrical impedance analysis (BIA)

144/2863

CORRELATION OF VISCERAL FAT LEVEL WITH
WAIST CIRCUMFERENCE IN PATIENTS WITH
NORMAL WEIGHT

**Background and objectives:** The aim of assessing nutritional status is to determine the general health of a patient from a nutritional point of view. Anthropometric techniques and bioelectrical impedance analysis (BIA) are two of the most widely used field methods due to their easy application, good reproducibility and low cost.

Knowing the size of visceral compartment allow us to predict
metabolic risks.

We aimed to correlate visceral fat level with waist circumfer-
ence in patients with normal weight.

**Methods:** We analyzed data from retrospective studies of patients undergoing nutritional assessment in the period 2015 by multifrequency BIA according to medical criteria. Anthropometry with 4 skinfold thickness and waist measurement was evaluated. The Spearman method was used to evaluate correlation between groups.

**Results:** 1470 multifrequency bioelectrical impedance were performed. 19% (281) patient with normal weight 72% (202) female, Media BMI 22.4 kg/m2, 22% (44) of female population had a % of total body fat ≥ 35% and a 25% (50) had visceral fat higher than normal limit.

28% (79) were male, Media BMI 22,1 kg/m2, 5,1% (4) had a
total body fat ≥ 25%.

Global Rho spearman = 0.38 p 0.0000. By groups Female Rho
spearman 0.64 p 0.000 and Male Rho spearman 0.73 p 0.0000

The Clinical Research and Bioethics Committee of the Hospital Italiano de Buenos Aires approved the study.

**Conclusions:** In all patients evaluated a weak linear relationship can be seen, concluding that waist circumference should not be used as a single method for an estimate of visceral fat in normal weight patient probably because it does not take into account height.

The visceral fat size was independent of anthropometric in-
dicators of adiposity such as BMI and waist circumference. Data
analyzed by sex, evidenced an increased correlation in women.

**Keywords:** Bioelectrical impedance analysis (BIA)

<footnote>(1)Venczel, Analía Verónica; (2)Cristina, Monica; (2)Yuma, María;</footnote>

<footnote>(2)Aragona, Hilda Susana; (3)Posadas Martines, Lourdes; (3)Perez,</footnote>

<footnote>Lucía; (3)Giunta, Diego; (2)Gutt, Susana.</footnote>

<footnote>(1)Nutritionist. Internal Medicine Service. Italian Hospital of Buenos Aires. Argentina; (2)Nutrition Section - Internal Medicine Service - Italian Hospital of Buenos Aires. Argentina; (3)Clinical Research Section - Internal Medicine Service - Italian Hospital of Buenos Aires.</footnote>

<footnote>(1)Gutt, Susana; (1)Cristina, Monica; (1)Yuma, María; (1)Aragona, Hilda Susana; (1)Schraier, Silvio; (2)Posadas Martines, Lourdes; (2)Perez, Lucía; (2)Giunta, Diego; (1)Venczel, Analía Verónica.</footnote>

<footnote>(1)Nutrition Section - Internal Medicine Service - Italian Hospital of Buenos Aires. Argentina; (2)Clinical Research Section - Internal Medicine Service - Italian Hospital of Buenos Aires.</footnote>
PREVALENCE OF MANLNUTRITION IN ENRICH
PROGRAM SITES OF BANGLADESH, KENYA, MY-
ANMAR, PAKISTAN AND TANZANIA

(1)Tolossa, Asrat; (2)Thomas, Abena; (3)Gillespie, Melanie; (4)Sellen, Daniel.

(1)MD, MPH, Chief of Party. ENRICH Program. World Vision Canada. Canada; (2)Grant Manager/DME advisor. ENRICH Program. World Vision Canada. Canada; (3)DME Team Lead. World Vision Canada. Canada; (4)Prof. Director The Joannah and Brian Lawson Centre for Child Nutrition. University of Toronto. Canada.

**Background and objectives:** ENRICH is a Maternal New-
born and Child Health and Nutrition program funded by the Gov-
ernment of Canada and implemented in Bangladesh, Pakistan,
Myanmar, Kenya and Tanzania. ENRICH will use nutrition path-
way to reduce U5M by strengthening health systems and improv-
ing basic nutrition and nutrition sensitive services and address-
ing nutrition-related causes such as: poor breastfeeding, stunting,
wasting, and vitamin A and zinc deficiencies (Lancet, 2013). One
of the strategies is promotion of production and consumption of
biofortified micronutrient-rich crops which help to reduce vita-
min A deficiency and improved iron status among CU5

**Methods:** The anthropometric nutrition assessment was done as part of baseline household survey of the ENRICH program. The sample size for each country was estimated using ENA for SMART software. A proportional stratified multi-stage cluster method was used to select respondents. Ethical approval was secured from University of Toronto and from the concerned authorities at country level. Anthropometric measurements were done among 6-59 months old children.

**Results:** Prevalence of stunting was 31.0% in Bangladesh, 33.1% in Kenya, 31.5% in Tanzania, 44.6% in Pakistan, and Myanmar 39%. The prevalence of wasting was 6.5%, 5.3%, 4.9%, 10.2% and 7.0% in Bangladesh, Kenya, Tanzania, Pakistan and Myanmar respectively. Under weight prevalence was also 20.8%, 17.8%, 15.3%, 29.7% and 28.2% in the same order. Poor infant and young child feeding practices, food insecurity and poor access to nutrition and health services were found to be some of the contributing factors. The proportion of severely food insecure households ranged from 11% to 32.3% among the five countries.

**Conclusions:** Malnutrition is a serious public health problem in ENRICH project sites of the implementing countries. Chronic food insecurity and poor infant and young child feeding practices are some of the factors contributing to the high level of malnutrition. Integrated nutrition specific and sensitive interventions which are effective in reaching the poor are required to address critical underlying determinants of malnutrition. Assessment of other local determinants of poor feeding practices and tailored interventions can also play a central role in preventing undernutrition.

**Keywords:** Stunting, Wasting, Underweight, Undernutrition

**Further collaborators:**

-Egerton University- Kenya. Aga-khan University- Pakistan.
BRAC University- Bangladesh. ENTAF - Tanzania. World Vision
East Asia Region - Myanmar.

THYROGLOBULIN IN CHILDREN WITH LONG-
TERM EXPOSURE OF IODINE EXCESS

(1)Zhang, Wanqi; (2)Chen, Wen; (2)Wang, Wei; (3)Hao, Yunmeng; (3)Wu, Yalan; (4)Bian, Jiancao; (2)Lin, Laixiang; (2)Tan, Long; (3)Shen, Jun;

(4)Wang, Xiaoming.

(1)Public Health College, Tianjin Medical University; (2)PhD. Department of Food nutrition and hygiene, Tianjin Medical University, Tianjin, China; (3)MD. Department of Food nutrition and hygiene, Tianjin Medical University, Tianjin, China; (4)The Shandong Institute for Endemic Disease Control and Research, Shandong, China.

**Background and objectives:** This study investigated thyroglobulin (Tg) concentrations in children with long-term exposure of iodine excess from water.

**Methods:** In a multi-stage cross-sectional survey, we collected, on two nonconsecutive days, two 24-h urine samples and determined 24-h urinary iodine excretion (24-h UIE, µg/d) and then calculated habitual daily iodine intake. Tg of children was detected.

**Results:** In this study, 2034 children had Tg measured including 1001 boys and 1033 girls. The water iodine content in this study was 181 µg/L (67.1-403) µg/L. The habitual daily iodine intake of children was 298 µg/d (188-436) µg/d. The median Tg concentration of children was 16.0 µg/L (9.8-26.1) µg/L in this study, and 237 (12%) children had Tg>40 µg/L while 85 (4.2%) children had Tg<4 µg/L. Tg was not associated with iodine intake concentrations up to the threshold value of 120 µg/d. When the iodine intake concentration was >120 µg/d, Tg was associated with increased iodine intake (β: 0.5; 95% CI: 0.4, 0.6; P<0.001). Taking iodine intake of 100-149 µg/d as the reference, logistic regression analysis for the association between iodine intake and Tg>40 µg/L, indicating that children from iodine intake range of ≥600 had higher risk of TGR compared to iodine intake of 100-149 µg/d (OR: 2.4 95% CI: 1.2, 4.9; P=0.018) while no differences were observed in other iodine intake ranges.

**Conclusions:** Tg concentrations of children who were exposed to high iodine intake were higher, however, the risk of Tg>40µg/L was not different among different iodine intake levels.

**Keywords:** Children, thyroglobulin (Tg), iodine excess

**Further collaborators:**

Section of Endocrinology, Diabetes, and Nutrition, Boston University School of Medicine, Boston, MA, USA (Elizabeth N Pearce, MD); the Human Nutrition Laboratory, Swiss Federal Institute of Technology, Zurich, Switzerland (Michael B Zimmermann, PhD)
# DETERMINATION OF IODINE CONTENT BY ICP-MS FOR IODINE BALANCE STUDY

Zhang, Wanqi; Cui, Tingkai; Shen, Jun; Pan, Ziyun; Tan, Long.

Department of Nutrition and Food Hygiene. School of Public Health. Tianjin Medical University. Tianjin. China.

**Background and objectives:** Iodine balance study on adults, neonates, pregnant and lactating women is essential to provide strong evidence for Dietary Reference Intakes (DRIs. Thus iodine content in a host of food, water, table salt, urine, breast milk and feces samples need to be analysed. Meanwhile demand for assessment of iodine content in serum and saliva samples is increasing due to the rising awareness of iodine status. Inductively Coupled Plasma Mass Spectrometry (ICP-MS) prevails in determining elements, including iodine, as its rapidity, low detection limits and minimal sample amount needed for analysis. This study aims to develop a fast, simple, and cheep method to determine iodine concentration in massive samples for iodine balance study and iodine status survey or evaluation.

**Methods:** Weigh 0.2g shredded food or feces samples and digest with 4.8mL 1% tetramethylammonium hydroxide (TMAH) in 5mL eppendof (EP) tube at 90°C for 24h, then transfer 0.5mL supernatant into 2mL EP tube after centrifuge at 3000 rpm for 15min, dilute with 1.5mL ultra-purified (UP) water, and finally centrifuge at 12000 rpm for 5min. Transfer 0.25mL breast milk samples into 5mL EP tube while with 2.75mL 0.5% ammonia at 90°C for 3h, then centrifuge at 3000 rpm for 15min and transfer 0.5mL supernatant into 2mL EP tube followed by diluted with 1.0mL UP water. Driking water, salt, serum, urine, and saliva samples are directly diluted with 7mmol/L ammonia. The final solution can be injected into ICP-MS for iodine analysing.

**Results:** The precision (RSD: 2.65%-8.85%) and recovery rate (88.08%-115.04%) of the method for assessment of above samples were satisfactory.

**Conclusions:** These sample treatments make iodine determination in a large number of samples fast, simple and cheep and it can be used for iodine status survey or evaluation, especially for iodine balance study.

**Keywords:** Iodine; determination; ICP-MS

# NEW INSIGHTS INTO THE PANDEMIC OF LOW VITAMIN "D" LEVELS AND ITS ASSOCIATION WITH SEMEN QUALITY AND HORMONAL LEVELS IN FERTILE AND INFERTILE MALE SUBJECTS

(1)Ciccone, Inari; (2)Pariz, Juliana; (3)Costa, Elaine; (4)Hallak, Jorge.

(1)MS Student. Androscience. Faculdade de Medicina da Universidade de São Paulo.; (2)PhD Student. Biomedical. Androscience. University of Sao Paulo Medical School FMUSP. Brazil; (3)Associate Professor of Endocrinology. Division of Endocrinology. Hospital das Clínicas of University of São Paulo. Brazil; (4)PhD. Andrologist. Associate Professor Of Urology. Department of Pathology. University of Sao Paulo Medical School. Androscience. Brazil.

**Background and objectives:** Vitamin D is a versatile signaling molecule, that targets also male reproductive organs, in addition to effects on bone, calcium and phosphate homeostasis. Accumulating evidence from animal and human studies suggests that it is involved in reproduction metabolism. Objective: To evaluate the influence of vitamin D status on semen quality and hormonal profile in male fertility and infertility.

**Methods:** We evaluated 100 men (aged 18 to 50 y.o.) from a private medical clinic. Infertility (n=70) and fertility (n=30) groups. According to vitamin D level status all of them were classified in sufficient (group 1 - 25OHvitD ≥ 30ng/ml), insufficient (group 2 - 25OHvitD from 21 to 29ng/ml) and deficient (group 3 - 25OHvitD ≤20ng/ml). Blood samples were collected to analyze serum vitamin D, LH, FSH, estradiol, total and free testosterone, prolactin and SHBG levels. Semen was analyzed according to WHO guidelines, strict criteria and sperm functional tests were performed (ROS, CK, beads). In addition, karyotype, frequency of varicocele, smoking, alcohol ingestion, and body composition were considered. Statistical analysis was performed by SPSS program version 19.0 (SPSS Inc., Chicago, IL). T-test was used for unpaired samples for statistical analysis. Statistical significance was considered with P value < 0.05.

**Results:** According to vitamin D status, patient distribution was: Infertile: group 1 - 28.5% (20/70), group 2 - 43% (30/70) and group 3 - 28,5% (20/70). Fertile: group 1 - 34% (10/30), group 2 - 40% (12/30) and group 3 - 26% (8/30). Sperm motility was significantly lower in group 3 in comparison to group 1 in infertile patients. Regarding fertile men, we found higher sperm volume in group 3 than the group 1, a significant reduction of sperm concentration and worse morphology by strict criteria in group 3 in comparison to group 1. Hormonal levels were similar in all vitamin D groups.

**Conclusions:** Our results demonstrated that sufficient vitamin D levels has a positive influence on spermatogenesis and semen quality, suggesting that vitamin D replacement should highly be concerned on male fertility treatment and that low vitamin D have reached epidemic proportions in industrialized cities.

**Keywords:** Nutrition and Male infertility. Male fertility treatment. 25OHvitaminD deficiency.

**Further collaborators:** Financial support: Androscience and CNPQ
# EVALUATION OF HEMOGLOBIN MEASUREMENT TOOLS FOR CHILD ANEMIA SCREENING IN RWANDA

(1)Parker, Megan E Parker; (2)Han, Zhen; (3)Abu-Haydar, Elizabeth;

(4)Matsiko, Eric; (4)Iyakaremye, Damien; (5)Tuyisenge, Lisine;

(6)Magaret, Amalia; (7)Lyambabaje, Alexandre.

(1)PATH; (2)BSc, PhD. Nutrition Innovation, PATH. USA.; (3)BSc, MPH. Devices & Tools, PATH. USA; (4)BSc, MSc. Human Nutrition and Dietetics Department, University of Rwanda, Kigali, Rwanda; (5)BSc, MD, MSc. Pediatrics. Centre Hospitalier Universitaire de Kigali, Kigali, Rwanda; (6)BSc, MSc, PhD. Departments of Lab Medicine and Biostatistics, University of Washington, Seattle, WA; (7)BSc, MSc, PhD. Mathematics and Applications. University of Rwanda, Kigali, Rwanda.

**Background and objectives:** Blood hemoglobin (Hb) level is a common indicator for diagnosing anemia and is often determined through laboratory analysis of a venous or capillary blood sample. Drawing blood samples, however, poses challenges in young children, increases the risk of infection and disease transmission, and is sometimes unacceptable to caregivers. One alternative to laboratory-based methods is use of the handheld HemoCue® Hb 201+ device, which requires a finger prick and wicking of blood into a pretreated cuvette for analysis. This wicking method is commonly used in clinic and community settings, but an alternative HemoCue® gravity method is being investigated for improved accuracy. Further, recent developments in noninvasive technologies could provide an accurate, rapid, safe, point-of-care option for Hb estimation while addressing some limitations of current tools, but the performance of these devices has not been assessed in low-resource settings. This study evaluated the performance of two HemoCue® Hb 201+ blood sampling methods and a noninvasive device (Pronto® with DCI-mini™ sensors) in a Rwandan pediatric outpatient clinic.

**Methods:** Reference Hb values were determined in 132 children 6 to 59 months of age by using a standard hematology analyzer (Sysmex KN21). 66 children were tested using the HemoCue® wicking method and 66 were tested using the HemoCue® gravity methods. 112 of the children had successful Hb readings with Pronto® DCI-mini™. Statistical analysis was used to assess the level of bias generated by each method and the key drivers of bias.

**Results:** The HemoCue® gravity method was the least biased compared with the HemoCue® wicking method and the Pronto® with DCI-mini™ sensors. The biases of the HemoCue® wicking and Pronto® DCI-mini™ methods were inversely related to the Sysmex KN21 results. Both HemoCue® sampling methods correctly classified patients' anemic status at or above 80%, while the Pronto® DCI-mini™ device had a correct classification rate of only 69%.

**Conclusions:** The HemoCue® gravity method was more accurate than the traditional HemoCue® wicking method in this study, but its accuracy and operational feasibility should be confirmed by future studies. The Pronto® DCI-mini™ devices showed considerable promise but require further improvements in sensitivity and specificity before wider adoption.

**Keywords:** Hemoglobin. HemoCue®, Noninvasive Device, Rwanda, Children

# DETERMINATION CONTENTS OF THE DIGESTIBLE INDISPENSABLE AMINO ACIDS (DIAAS) OF NINE KINDS OF GRAIN CEREAL USING RAT MODEL

(1)Han, Fei; (1)Han, Fei; (2)Han, Fenli; (3)Chen, Xi; (3)Song, Ge; (4)Fan, Liuping.

(1)PhD. Nutritionist. Academy of State Administration of Grain. P.R.China; (2)Master. Food College. Jiang Nan University. P.R.China; (3)Master. Academy of State Administration of Grain. P.R.China; (4)PhD. Nutritionist. Food College. Jiang Nan University. P.R.China.

**Background and objectives:** Recently, for the grains, most of the researches are mainly focus on the antioxidation, prevention of chronic diseases and processed food as well as its nutritional compositions. However, there are few researches about the grain amino acid in vivo digestibility and formula design with digestible indispensable amino acid (DIAA) of grains. This research studies the contents of the digestible indispensable amino acids (DIAAS) of the nine typical grain cereals from different provinces in China.

**Methods:** The research has been made on the impacts of the protein sources in such nine kinds of grains on the apparent, true ileal amino acid digestibility and DIAA using rat model. To prepare diets using 9 grains for atmospheric pressure cooked that meet the nutritional needs of rats according to NRC requirements.

**Results:** The results showing that the average total apparent ileal amino acid digestibility values in the whole wheat and coix groups are significantly higher than other grain groups (P < 0.05) and the ileal true amino acid digestibility and ileal apparent amino acid digestibility values are characterized by a basically consistent trend; the available lysine digestibility is highest in the whole wheat and millet groups but lowest in the tartary buckwheat group and there is no significant difference in others groups (P > 0.05). Moreover, the research has been made on the ileal DIAA and available lysine contents of the protein sources in different grains, with the results showing that the digestible lysine contents in the oats and rice groups are higher than the millet, foxtail millet, coix and whole wheat groups but lower than the common and tartary buckwheat groups (P < 0.05) and the amino acids in legumins are more easily digestible than that in prolamins.

**Conclusions:** The nutritional value of the 9 grains was further evaluated by digestible indispensable amino acid score (DIAAS), the order were as follows: buckwheat group>tartary buckwheat group>oat group>brown rice group>polished rice group>whole wheat group>coix group>millet group>proco-millet group.

**Keywords:** grain cereals; protein; digestible indispensable amino acids (DIAAS)
## 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.

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## 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.

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## 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

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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 contains 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

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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

**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

**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.

<footnote>(¹)Iwaniak, Anna; (²)Darewicz, Malgorzata; (³)WOŁOWSKA, Ewelina;</footnote>

<footnote>(²)Minkiewicz, Piotr.</footnote>

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

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

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

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

<footnote>(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.</footnote>
**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 with experimentally confirmed bioactivities as well as calculation functions helping to evaluate the protein value as the source of biopeptides.

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.

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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.

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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-
ses 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

**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

**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.

<footnote>(1)Aranha, Flávia; (2)Barretto, Daniela; (2)Burriguel, Gabriela;</footnote>

<footnote>(2)Mendes, Juliana; (2)Guidorizi, Luana; (2)Barchi, Natalia.</footnote>

<footnote>(1)1PhD. 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.</footnote>

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

<footnote>(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.</footnote>
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.
**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

**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.

144/845

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-

ponents 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.

144/965

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 part-ly 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

**Background and objectives:** Sphingolipids are ubiquitous in all eukaryotic organisms and have attracted attention as physio-logically functional lipids. Recently, various physiological func-tions of dietary sphingolipids, especially improving the skin bar-rier 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 evalu-ated 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 charac-terized 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

**Background and objectives:** We evaluated the effect of evi-dence-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.

<footnote>(1) Sugawara, Tatsuya; (2) Tomonaga, Nami; (3) Duan, Jingjin;</footnote>

<footnote>(4) Manabe, Yuki; (5) Tsuduki, Tsuyoshi; (6) Hirata, Takashi.</footnote>

<footnote>(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.</footnote>

<footnote>(1) Kosehira, Marie; (2) Kitaichi, Nobuyoshi.</footnote>

<footnote>(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.</footnote>
# 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.

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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 Bioherium 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.

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## RED PITAYA PEEL EXTRACT IMPROVED STEATOSIS AND INFLAMMATION IN ALCOHOLIC LIVER DISEASE

(1)Yang, Hsin-Yi; (2)Yeh, Wan Ju; (3)Tsai, Chia-Chun; (2)Chen, Jiun-Rong.

(1)PhD. Department of Nutrition, I-Shou University. Taiwan.;

(2)PhD. School of Nutrition and Health Sciences. Taipei Medical University. Taiwan.; (3)MSc. School of Nutrition and Health Sciences. Taipei Medical University. Taiwan.

**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.

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## FORMULATION AND CHARACTERIZATION OF A CEREAL BAR AS A FUNCTIONAL FOOD

(1)Vallejo Trejo, Lucia; (2)Vallejo Trejo, Lucia.

(1)mg lic Lucía Vallejo Trejo sistema provincial de salud; (2)San Miguel de Tucuman.

**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.
# 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.

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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-

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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.

## FOOD SUPPLEMENTS BASED ON COMPOUNDS FROM THE TRADITIONAL NUTRITION IN BULGARIA

Handjiev, Svetoslav.

MD, Bulgarian association for the study of obesity and related diseases (BASORD).

**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.

## ENZYMATICALLY-SYNTHESIZED GLYCOGEN INHIBITS INFLAMMATORY RESPONSE IN ANTI-GEN-INDUCED BASOPHILIC AND MAST CELLS

(1)Yoshioka, Yasukiyo; (2)Furuyashiki, Takashi; (3)Ashida, Hitoshi.

(1)PhD. Postdoctoral fellow. Graduate School of Science, Technology and Innovation. Kobe University, Japan.; (2)MS. Scientist. Institute of Health Sciences. Ezaki Glico Co. Ltd. Japan;

(3)Professor. Graduate School of Agricultural Science. Kobe University, Japan.

**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.