# EFFECT OF CINNAMON ON MORPHOLOGY AND PHYSIOLOGY OF LIVER IN RATS

Safdar, Mahpara.

PhD. Department of Home & Health Sciences, Allama Iqbal Open University.

**Background and objectives:** The study on cinnamon was initiated to assess its overall effect on morphology and physiology of liver in rats and to extrapolate the results to humans.

**Methods:** The study was conducted in the department of Human Nutrition, Agricultural University, Peshawar, Pakistan. Seven two female albino rats weighing 180-270g were divided into groups I, II and III, each having 24 rats. Group I was for placebo, II for 4mg and III for 12 mg cinnamon per day respectively. The total duration of the experiment was 60 days, out of which, 40 days were for cinnamon and the last 20 days were without cinnamon. Six rats from each group were sacrificed at day 0 to get the base line data for all groups prior to the start of experiment. Six rats from each group were sacrificed at days 20, 40 and 60 for obtaining blood sample for biochemical analysis and removing liver for viewing structural changes. Liver functions namely bilirubin, alkaline phosphatase (AP), alanine transaminase (ALT), albumin, globulin, albumin to globulin (A/G) ratio were determined by using standard methods. The changes in cellular structure of liver were viewed under Olympus BX51 microscope.

**Results:** Cinnamon did not significantly change the concentrations of AP, ALT, globulin and A/G ratio throughout the study period. Bilirubin was non-significantly (P>0.05) changed at day 20 but significantly (P<0.05) increased at days 40 and 60, as compared to the values at day zero. Also albumin was non-significantly changed at day 20 but significantly increased at day 40 but non-significant at day 60, as compared to the value of albumin at day zero.

The cellular structure of liver was normal showing normal portal tract, bile duct surrounded by cords and trabeculae of hepatocytes with intervening sinuses and kupffer cells up to day 20, but mild inflammation, showing increased lymphocytes was observed at days 40 and 60.

**Conclusions:** In the light of this study, if results of rats are valid for human, then cinnamon safety is doubtful for liver physiology and morphology. It is suggested that further studies may be conducted to verify the safety of cinnamon intake.

**Keywords:** Cinnamon safety, Morphology, Physiology, Liver.

**Conflict of Interest Disclosure:** Sindhu Mathew, T. Emilia Abraham, Khan A, Rui wang, Ruijiang wang.

**Further collaborators.**

Food Chemistry

Journal of Food Process Engineer

# SOCIAL JET LAG, OBESITY AND NON-COMMUNICABLE CHRONIC DISEASES

(1)Mota, Maria; (2)Silva, Catarina; (3)Balieiro, Laura; (4)Fernandes, Bruna;

(5)Rocha, Jokasta; (6)Furlan, Luisa; (7)Fahmy, Walid; (8)Crispim, Cibele.

(1)Federal University of Uberlândia. Uberlândia. Minas Gerais. Brazil; (2)PhD Student. Nutritionist. Federal University of Uberlândia, Uberlândia, Minas Gerais. Brazil.; (3)PhD Student. Nutritionist. Federal University of Uberlândia, Uberlândia, Minas Gerais Brazil; (4)Nutritionist student. Federal University of Uberlândia, Uberlândia, Minas Gerais Brazil; (5)Nutritionist. Federal University of Uberlândia, Uberlândia, Minas Gerais Brazil; (6)Master Degree student. Nutritionist. Federal University of Uberlândia, Uberlândia, Minas Gerais Brazil; (7)Master Degree. Hospital e Maternidade Municipal de Uberlândia, Department of Obstetrics, Uberlândia, Brazil; (8)PhD. Nutritionist. Federal University of Uberlândia, Uberlândia, Minas Gerais Brazil.

**Background and objectives:** Studies have suggested that disruption of the circadian system, such social jetlag (a measure of the difference between sleep times on work days and days off) may lead to obesity and influence metabolic parameters. Although social jetlag have already been associated with overweight in other populations, little is known about the associate of these factors with chronic diseases commonly related to obesity. The objective of this study was to analyze the association of social jetlag between anthropometric and obesity status among individuals with chronic diseases.

**Methods:** The study was cross-sectional using volunteers [n=792, 581 (73%) female, average age 55.9 ± 12.4] with non-communicable chronic diseases: obesity, systemic arterial hypertension, type 2 diabetes mellitus or dyslipidaemia; who were attended at outpatient clinics of the public health service of the city of Uberlândia-MG (Brazil). The volunteers answered a questionnaire that assessed: demographic, physical activity and habitual sleep pattern; and anthropometric characteristics [height, weight, body mass index (BMI) and waist circumference] were determined. From the medical records of the volunteer clinical and biochemical information was collected. Obesity status has been classified in three levels: non-obese: BMI<30 kg/m2; healthy obese: BMI ≥ 30 kg/m2 and less than three high-risk biomarkers for metabolic syndrome; and unhealthy obese: BMI ≥ 30 kg/m2 and high-risk values on three or more biomarkers for metabolic syndrome. Social jetlag was calculated based on the absolute difference between mid-sleep time on weekends and weekdays.

**Results:** Multivariate logistic regression adjusted for possible confounding variables indicated a higher risk of being overweight (BMI > 25 kg/m2) for individuals that presented social jetlag (>1 h) (odds ratio [OR]= 2.0, confidence intervals [CI] = 1.2-3.6, p=0.006]; and a risk of being an unhealthy obese (OR=1.8, CI=1.1-2.8, p=0.01) for individuals that presented social jetlag (>1 h) in comparison with those without social jetlag (<1 h).

**Conclusions:** Social jetlag is associated with higher odds of being overweight and unhealthy obese, confirming that circadian misalignment may predispose improper weight gain and metabolic syndrome.

**Keywords:** Social jetlag, Sleep, Obesity, Overweight, Unhealthy obesity.
# RECOVERY, RELAPSE, AND EPISODES OF DEFAULT IN THE MANAGEMENT OF ACUTE MALNUTRITION IN CHILDREN IN HUMANITARIAN EMERGENCIES: A SYSTEMATIC REVIEW

(1)Akparibo, Robert; (2)Booth, Andrew; (3)Lee, Andrew C.K.

(1)BSc, MSc, MPH, PhD. School of Health and Related Research. University of Sheffield. UK; (2)BA, Dip Lib, MSc, PhD. School of Health and Related Research. University of Sheffield. UK.; (3)MB ChB, MSc, MD. School of Health and Related Research. University of Sheffield. UK.

**Background and objectives:** This systematic review, commissioned by the Humanitarian Evidence Programme (HEP) and carried out by a research team from the University of Sheffield, represents the first attempt to apply systematic review methodology to establish the relationships between recovery and relapse and between default rates and repeated episodes of default or relapse in the management of acute malnutrition in children in humanitarian emergencies in low- and middle-income countries.

**Methods:** Quantitative, mixed methods academic literature and grey published programme reports were identified and reviewed. Peer-reviewed literature search was conducted in 14 academic databases. Grey literature, including programme reports, were searched in Google Scholar, websites of relevant organisations, and by engaging with stakeholders in the nutrition field. Studies published prior to 1980 were excluded, as were any papers not published in English.

**Results:** A total of 9,574 articles, studies and programme reports relating to acute malnutrition were retrieved from the searches conducted. Following the removal of duplicates, screening and quality appraisal, 24 articles and reports were eligible for review. 23 of these focused on sub-Saharan Africa: eight were conducted in Malawi, 5 in Ethiopia, 3 in Niger, 3 in Sudan and the remaining 4 in Angola, Chad, Kenya and Sierra Leone. One study was conducted in Afghanistan. Most studies and programme reports reported on quantitative outcomes and only two contained both quantitative and qualitative outcomes. The 22 quantitative studies included 8 clinical efficacy and effectiveness trials using randomized controlled designs, 7 observational cohort studies, and 7 programme evaluation reports. Only 6 of the 24 studies included in this review addressed the issue of relapse and/or reported relapse rates. None of the studies addressed the relationship between relapse and default or return default, and little evidence was found on the long-term impact of programmes implemented to manage MAM and SAM in emergencies.

**Conclusions:** This review provides further confirmation that RUTF used in an outpatient setting is effective at promoting recovery from SAM and reducing mortality. It could not be established whether default rates reported were lower according to the WHO 2013 protocol. Data relating to relapse is limited and need further research.

**Keywords:** Acute malnutrition, Children, Humanitarian, Emergency, Systematic Review.

**Further collaborators.** DFID/Oxfam GB, Feinstein International, UKAiD, Erin Boyd, Mark Manary and Seth Adu-Afarwuah.

# MOLECULAR BASES UNDERLYING THE CHEMO PREVENTIVE EFFECTS OF DOCOSAHEXAENOIC ACID

(1)Ajami, Marjan; (2)Pazoki Toroudi, Hamidreza.

(1)PhD. Nutritionist. Dept. of Food and Nutrition Policy and Planning Research, National Nutrition and Food Technology Research Institute, Faculty of Nutrition Sciences and Food Technology, Shahid Beheshti University of Medical Sciences, Tehran, Iran; (2)Associate Professor of Physiology/ Dept. of Physiology,Iran University of Medical Sciences.

**Background and objectives:** Colorectal Cancer (CRC) is the third most commonly cancer around the world. Nutraceutical are food products like drug which share health benefits in prevention and treatment of many diseases. The nutraceuticals like omega-3 Docosahexaenoic acid may have chemopreventive activities Docosahexaenoic acid (DHA) is an omega-3 fatty acid and it may be considered as potential therapeutic agent for colon cancer.

**Methods:** Colon cancer was induced by injecting 6-8-week-old, rats weighing 200-250 g (Pasteur Institute of Iran, Tehran) with 10 mg/kg 1,2-dimethylhydrazine (DMH) twice per week for 12 weeks. 26 male Wistar rats divided in to two groups. DHA group treated with 400 mg/kg DHA (n=13). VEGF (Vascular endothelial growth factor) and mTOR levels in colorectal cancer were examined by Western blotting.

**Results:** DHA reduced VEGF expression by 34.3 ± 1.7 vs. control group 71.2±3.4 (Protein expression/b-actin expression). DHA reduced mTOR expression by 32.1 ± 1.2 vs. control group 65.2±3.8 (Protein expression/b-actin expression).

**Conclusions:** mTOR dysregulation has a vital role to play in many types of cancers. The current study suggested that DHA is able to inhibit VEGF and mTOR expression in colon cancer cells and DHA is a potent therapeutic agent that can inhibit tumor growth and angiogenesis in vivo by decreased phosphorylation of mTOR and subsequently the blockage of VEGF secretion in tumor cells pathway.

**Keywords:** mTOR, VEGF, DHA, Colon cancer.
# THE ASSOCIATION BETWEEN MINDFULNESS AND THE METABOLIC SYNDROME IS MODIFIED BY HISTORY OF DEPRESSION

(1)Guyot, Erika; (2)Baudry, Julia; (3)Allès, Benjamin; (4)Hercberg, Serge;

(5)Galan, Pilar; (6)Kesse-Guyot, Emmanuelle; (6)Péneau, Sandrine.

(1)MSc student. Equipe de Recherche en Epidémiologie Nutritionnelle. Centre de Recherche en Epidémiologie et Statistiques, Inserm (U1153), Inra (U1125), Cnam, Université Paris 13, COMUE Sorbonne Paris Cité. France.; (2)PhD student. Equipe de Recherche en Epidémiologie Nutritionnelle. Centre de Recherche en Epidémiologie et Statistiques, Inserm (U1153), Inra (U1125), Cnam, Université Paris 13, COMUE Sorbonne Paris Cité. France;

(3)Equipe de Recherche en Epidémiologie Nutritionnelle, Centre de Recherche en Epidémiologie et Statistiques, Inserm (U1153), Inra (U1125), Cnam, Université Paris 13, COMUE Sorbonne Paris Cité, Bobigny. France.; (4)Prof. Equipe de Recherche en Epidémiologie Nutritionnelle. Centre de Recherche en Epidémiologie et Statistiques, Inserm (U1153), Inra (U1125), Cnam, Université Paris 13, COMUE Sorbonne Paris Cité. France.; (5)PhD. Equipe de Recherche en Epidémiologie Nutritionnelle, Centre de Recherche en Epidémiologie et Statistiques, Inserm (U1153), Inra (U1125), Cnam, Université Paris 13, COMUE Sorbonne Paris Cité, Bobigny. France.; (6)PhD. Equipe de Recherche en Epidémiologie Nutritionnelle. Centre de Recherche en Epidémiologie et Statistiques, Inserm (U1153), Inra (U1125), Cnam, Université Paris 13, COMUE Sorbonne Paris Cité. France.

**Background and objectives:** The Metabolic syndrome (MetS) is a major public health burden since it predisposes to cardiovascular diseases, which are leading causes of death in developed countries. In addition to lifestyle habits, specific psychological characteristics have been shown to be associated with the MetS. Dispositional mindfulness, which is defined as non-judgmental awareness of the present moment, has recently been associated with eating disorders and overweight and could therefore be associated with the MetS. We aimed to examine in a cross sectional design the relationship between mindfulness and the MetS in a large sample of the adult general population and the influence of history of depression on this association.

**Methods:** A total of 17,490 adults participating in the Nutri-Net-Santé study who had completed the Five Facets Mindfulness Questionnaire and attended a clinical and biological examination were included. Multivariable logistic regression models adjusted for socio-demographic and lifestyle factors were performed to assess the relationship between mindfulness (and its subscales) and the MetS and cardiovascular risk factors.

**Results:** Among individuals with a history of depression, those with higher mindfulness were less likely to have a MetS (OR: 0.73, 95% CI: 0.57-0.93), a high waist circumference (OR: 0.79, 95% CI: 0.67-0.95), a low HDL-cholesterol level (OR: 0.73, 95% CI 0.55-0.95) and a high fasting blood glucose level (OR: 0.69, 95% CI: 0.54-0.89) (all p < 0.05). In those without history of depression, individuals with higher mindfulness were less likely to have a high waist circumference (OR: 0.89, 95% CI 0.82-0.97) (p < 0.01).

**Conclusions:** These findings support the importance to take individual psychological factors into account, and in particular mindfulness, in relation with the MetS, especially in people with a history of depression.

**Keywords:** Mindfulness, metabolic syndrome x, risk factors, depression.

**Conflict of Interest Disclosure:** Erika Guyot, Julia Baudry, Serge Hercberg, Pilar Galan, Emmanuelle Kesse-Guyot, Sandrine Péneau report no conflict of interest.

# DIETARY INTAKE OF NUTRIENTS AND COMPROMISED PERIODONTAL HEALTH: THE CONCORD HEALTH AND AGEING MEN PROJECT

(1)Milledge, Kate; (2)Cumming, Robert; (3)Wright, Frederick;

(4)Naganathan, Vasikaran; (5)Blyth, Fiona; (6)Le Couteur, David;

(7)Hirani, Vasant.

(1)Dietitian. School of Life and Environmental Sciences. University of Sydney. Australia; (2)PhD. Professor of Epidemiology. School of Public Health. University of Sydney. Australia.; (3)PhD. Clinical Professor. Centre for Education and Research in Ageing. Concord Repatriation General Hospital. University of Sydney. Australia;

(4)PhD. Associate Professor. Centre for Education and Research in Ageing. Concord Repatriation General Hospital. University of Sydney. Australia.; (5)PhD. Associate Dean. Sydney Medical School. University of Sydney. Australia.; (6)PhD. Professor of Geriatric Medicine. ANZAC Research Institute. Charles Perkins Centre. University of Sydney. Australia.; (7)PhD. Senior Lecturer. School of Life and Environmental Sciences. University of Sydney. Australia.

**Background and objectives:** More than half of Australian adults aged 65 years and older have moderate to severe periodontal disease. Yet in the older population, research specifically examining the association between diet and periodontal health is limited. The aim of this study was to examine the association between dietary intake of nutrients and periodontal health in older men participating in the Concord Health and Ageing Men Project (CHAMP) study.

**Methods:** A preliminary analysis was conducted on the 294 men, (mean age: 84 years, SD: ±3.7 years), who had completed a standardised validated dietary assessment and had a comprehensive oral health examination during the CHAMP 8-year follow up. Having three or more teeth with at least one site with a Probing Depth (PD) ≥3mm, or at least one site with an Attachment Loss (AL) of ≥5mm were used as indicators of compromised periodontal health. Nutrients were categorised into quartiles and logistic regression was used for all statistical analysis.

**Results:** The prevalence of compromised periodontal health was high, with 64% of men had three or more teeth with at least one site that had a PD ≥ 3mm, while 85% of men had three or more teeth with at least one site that had an AL ≥ 5mm. After adjustment for potential confounders, men in the lowest quartile of percentage of energy from protein intake were much more likely than those in the highest quartile to have PD ≥ 3mm: OR 2.59(95% CI:
1.14 – 5.88). Men in the lowest quartile of folate intake were much less likely to have AL ≥ 5mm: OR 0.35(95% CI 0.12 – 0.99). The mean percentage of energy from protein intake was 18.4% (SD: ± 3.7%), and the mean intake of folate was 407.1µg (SD: ± 202.1µg). Beef was the main food source contributing to protein intake and vegemite was the main food source contributing to folate intake.

**Conclusions:** These preliminary results highlight that there is a complex association between the dietary intake of nutrients and the periodontal health of older men.

**Keywords:** Nutrition. Periodontitis. Diet. Older adults.

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IMPROVING THE MANAGEMENT OF SEVERE ACUTE MALNUTRITION AMONG INFANTS AND CHILDREN THROUGH CAPACITY BUILDING: FINDINGS FROM AN EVALUATION STUDY WITH MALNUTRITION ELEARNING

(1)Choi, Sunhea; (2)Yuen, Ho-Ming; (3)Annan, Reginald; (4)Monroy Valle, Michele; (5)Pulman, Andrew; (6)Aduku, Nana Esi; (7)Kyei-boateng, Samuel; (7)Pickup, Trevor.

(1)University of Southampton; (2)MSc. Lecturer and Senior Statistician. Faculty of Medicine, University of Southampton, Southampton, United Kingdom; (3)PhD. Lecturer and Nutritionist. Department of Biochemistry and Biotechnology. Kwame Nkrumah University of Science and Technology. Ghana.; (4)ND and MEd. Nutritionist. FUniversidad Rafael Landívar. Guatemala; (5)PhD. Researcher. Faculty of Medicine, University of Southampton, Southampton, United Kingdom; (6)BSc. Nutritionist. Department of Biochemistry and Biotechnology. Kwame Nkrumah University of Science and Technology. Ghana.; (7)BSc (Econ). Researcher. Faculty of Medicine, University of Southampton, Southampton, United Kingdom.

**Background and objectives:** The University of Southampton and International Malnutrition Task Force developed Malnutrition eLearning to reduce child mortality by Severe Acute Malnutrition (SAM) through training health professionals globally. Since made available in 2010, over 14,000 from 100+ countries used the course. To investigate its effectiveness, a 2-year evaluation study was conducted from 2015, face-to-face in Ghana and Central America (CA), and online in other countries.

**Methods:** Using a mixed method approach, the study explored if and how Malnutrition eLearning supported knowledge gain and behavioural change (application of knowledge in clinical practice), and resulting clinical outcomes in the management of SAM. Assessments, questionnaires and interviews/focus groups were conducted with individual in-service and pre-service participants pre- and post-training, and 12 months of medical records data collection, observations and hospital personnel interviews were carried out from participating healthcare institutions.

**Results:** Total 1,261 health professionals (Ghana:915, CA:142, other countries:201), and 10 hospitals and 2 community health centres in Ghana and 2 hospitals in CA participated in the study.

3,955 (pre:01/08/2014–31/07/2015) and 3,737 (post:01/08/2015–31/07/2016) medical records of children (0-60 months) were collected from the hospitals, and summary data on malnutrition cases (pre:76, post:67) from community health centres.

Individual participants scored significantly higher in the post assessment (mean difference(SD): 14.0(12.5), 95%CI(12.7, 15.2), p<0.001). 87% of in-service health professionals (102/117) applied their knowledge and changed clinical practice in screening, assessment, diagnosis and management of SAM. This group demonstrated retained knowledge 6-month after the training (mean difference from pre-assessment(SD): 12.7(11.7), 95%CI(10.4, 15.0),p<0.001). Significant increases (p<0.001) in recording malnutrition-markers, i.e. length/height and weight-for-length/height z-score, and diagnosed SAM cases (pre: 491(12.4%), post: 810(21.7%)) were observed. Mortality by SAM was declined from 26(5.9%) to 14(1.9%) (p<0.001). The community centres initiated the management of SAM (pre:0/32, post:7/21).

**Conclusions:** The results suggest that Malnutrition eLearning is effective in training the WHO guideline on the management of SAM. After a 2-day self-directed training with Malnutrition eLearning, the participants not only gained knowledge but were also able to apply the knowledge in their clinical practice, leading to significant impacts on clinical outcomes.

**Acknowledgement:** This study was supported by the UK Department for International Development Nutrition Embedding Evaluation Programme, managed by PATH.

**Keywords:** Management of Severe Acute Malnutrition, capacity building, nutrition sensitive intervention, eLearning, WHO guidelines on SAM management.

**Further collaborators:** Carmen Isabel Velásquez de West, Carmen Portillo, Andrew Penn, Sally Hickman.

144/3017

EFFECTS OF BIOTIN STATUS ON GLYCEMIC CONTROL IN RATS

Wang, Zhu; Xiang, Xuesong; Wei, Jiuling; Zhang, Xuesong.

National Institute for Nutrition and Health. Beijing. China

**Background and objectives:** Biotin, as a coenzyme of carboxylases, plays an important role in the process of neoglycogenesis, amino acid decomposition, fatty acid synthesis and energy metabolism in humans and mammals. In recent studies, it has showed that large dose of biotin supplement can be taken as drug to treat hyperglycemia, through potential mechanism of mediating gene regulation of histone biotinylation. The main source of body biotin are main from dietary intake and intestinal bacterial synthesizing, so biotin deficiency is rare, the relationship between biotin status and glycemic control should be studied.

**Methods:** 60 healthy SD rats (F/M half to half) weighting (100±10) grams were fed with non-biotin diet with raw egg white replacing casein to induce biotin deficiency model (NB), then divided into 6 test groups, daily oral given 0, 2, 4, 10 µg biotin, 1g resistance dextrin (RD, as a stimulant of intestinal source) or mix-
# WHAT IS NUTRITIONAL STATUS? PERSPECTIVE OF WOMEN IN IBADAN NIGERIA

(1)Okekunle, Akinkunmi Paul; (2)Omotosho, Melefa Moses; (3)Odok, Godwin Etta; (4)Akpa, Onoja Matthew; (5)Akinyele, Isaac Olaoluwa.

(1)Doctoral Student. Department of Nutrition and Food Hygiene. Harbin Medical University. China.; (2)BA. MA. PhD. Lecturer. Department of English and Literary Studies. University of Nigeria Nsukka. Nigeria.; (3)BSc. MSc. PhD. Lecturer. Department of Sociology. Federal University Dutsin-Ma Katsina. Nigeria.; (4)BSc. MSc. PhD. Senior Lecturer. Department of Epidemiology and Medical Statistics. University of Ibadan. Nigeria.; (5)BSc. MSc. PhD. Professor of Nutrition. Department of Human Nutrition. University of Ibadan. Nigeria.

**Background and objectives:** Good understanding of nutrition is fundamental to adequate nutritional status, health and well-being. In spite of several efforts to address malnutrition in Nigeria, the burden of poor nutritional status remains largely unfettered, especially among women. The burden obesity and underweight is prevalent in urban and rural settings respectively. This study explores the perspectives and understanding of nutritional status among women in Ibadan, Nigeria using qualitative approaches.

**Methods:** Four (4) focus groups were conducted among 27 women (15-49 years) identified by a multi-recruitment strategy from 285 households in urban and rural areas of Ibadan Nigeria, after duly informed consent. Using a predefined protocol, discussants' responses were explored with semi-structured but moderator-led discussions centred on 3 main themes; understanding of nutritional status, the type of food eaten to enhance good nutritional status and what major factor influences food choices in households. Responses were digitally recorded and professionally transcribed; all discussion transcripts were checked for accuracy and completeness, edited before analysis to directly include content pertaining to the themes and analysed using team-based qualitative analysis methods.

**Results:** Though discussant responses varied by age and region of residence, most discussants' consider the type of body shape they possess and avoidance of foods made up of fat as their indicator for good nutritional status. In addition, while young women and those residing in urban neighbourhoods consider taste and appetite, older and married women consider their spouse's food choices and stage of growth as a major factor that influences their food choices. The economic power of household is a major motivation that informs food choices of discussants and their households. Furthermore, most discussants were unfamiliar with the concept of food intakes and its implication on their nutritional status.

**Conclusions:** Nutrition education intervention strategies that integrate adequate management of monetary resources to promote consumption of right combination of foods is indispensable in enhancing the good nutritional status of women.

**Keywords:** Nutritional status, Women, Focus Discussion, Nigeria.

# MULTIFORTIFICATION OF BOUILLON IN CENTRAL AND WEST AFRICA: FEASIBILITY & IMPACT

(1)Habeych, Edwin; (2)Galaffu, Nicola; (2)Detzel, Patrick; (3)Prieto Patron, Alberto; (4)Messier, Marie-Chantal; (5)Duboc, Philippe.

(1)PhD. Scientist. Nestlé Research Center. Switzerland.; (2)PhD. Scientist. Nestlé Research Center. Lausanne. Switzerland.; (3)PhD. Scientist. Nestlé Research Center. Lausanne. Switzerland.; (4)MSc. Nutritionist. Nestec S.A. Vevey. Switzerland.; (5)PhD. Scientist. Nestlé Product Technology Center-Singen. Singen. Germany.

**Background and objectives:** Up to 38% of schoolchildren in West Africa suffer from goiter and iron deficiency anemia, which leads to early mortality, impairment of physical/cognitive development and poor school performance. Condiments such as bouillon are regularly use by families across the region to enhance the taste of home-cooked meals, making this product a useful vehicle for delivering micronutrients. Homogeneous, stable, and impactful multifortification bouillon is challenging as color, aroma, and flavor of the condiment can easily change during manufacturing, shelf-life and consumption.

The aim of this work is to develop a stable multifortified (iron and iodine) Maggi® bouillon cubes for industrial scale production and commercialization in Central and West Africa and assess its impact on the reduction of micronutrient deficiencies.

**Methods:** Stability of multifortified Maggi® bouillon cubes was extensively studied through assessment of color, aroma, flavor, and texture of the condiment during manufacturing, shelf-life, and consumption. Local preparations rich in polyphenols were tested to ensure stability. Process adaptation and optimization for proper micronutrients homogeneity were implemented. Nutritional impact was assessed through the development of a statistical model based on the principles of the Global Burden of Disease using nutritional data from Ivory Coast.

**Results:** Commercialization of stable multifortified Maggi® bouillon cubes was accomplished in 2012 as a mean to complement mass fortification programs. Key points for its industrializations and acceptability by consumers were: 1) product stability and sensory properties in traditional African recipes; 2) safety and nutritional relevance; 3) ensuring micronutrient homogeneity and safety; 4) no price increased. Multifortified Maggi® bouillon cubes was estimated in Ivory Coast to reduce by 4.7% the economic burden and 5.7% health burden in DALYs.

**Conclusions:** Multifortification of Maggi® bouillon cubes is an effective approach to reduce malnutrition in Ivory Coast (reducing by 4.7% the economic burden and 5.7% health burden in DALYs). Its implementation was challenging as stability, homogeneity, safety, cost, consumer acceptance, and nutritional impact were critical to ensure successful acceptance by consumers. Challenges going forward includes: bioavailability and density of micronutrients

**Keywords:** Fortification, Micronutrients, Bouillon, Public health.
COMPARISON OF OBESITY STATUS OF SCHOOL CHILDREN BY BODY MASS INDEX AND BY BODY COMPOSITION USING A STABLE ISOTOPE DILUTION TECHNIQUE IN NAIROBI CITY COUNTY, KENYA

(1)Kigaru, Dorcus Mbithe; (2)Loechl, Cornelia; (3)Ndung'u, Zipporah W.

(1)PhD. Nutritionist. Department of Foods, Nutrition & Dietetics. Kenyatta University. Kenya; (2)PhD. Department of Nuclear Sciences and Application. Division of Human Health. Nutritional and Health-Related Environmental Studies Section. International Atomic Energy Agency. Vienna. Austria.; (3)MSc. Dept of Food Science and Technology. Jomo Kenyatta University of Agriculture and Technology. Kenya.

**Background and objectives:** Changes in dietary intake, lifestyle and low physical activity contribute to obesity, the new silent killer which increases the risk of non-communicable diseases that threaten to burden already overstretched health services. Accurate and reliable indicators are required for early detection of childhood obesity and related health risks, for optimal nutrition intervention planning, monitoring and informing policy. Body Mass Index (BMI)-for-Age Z scores is widely used in epidemiological studies and screening for childhood obesity, however, it does not distinguish between body fat free mass and fat mass. Body fat accumulation and distribution is a better indicator of the risk of obesity and related diseases. This study assessed the accuracy of the BMI-based WHO classification of obesity for defining children as excessively fat by comparing it to the measure of body fatness using the criterion stable isotope method for body composition in school children in Nairobi City.

**Methods:** A cross-sectional study was conducted among 202 schoolchildren aged 8-11 year systematically sampled. A structured questionnaire was used to collect data on demographics, perceptions and knowledge about healthy habits, eating patterns, and physical activity. Anthropometry (BMI-for-Age Z scores) and body composition (fat mass and fat-free mass by deuterium dilution technique) was used to assess body fatness. Data was analysed using SPSS at significance p<0.05 at 95%CI.

**Results:** The prevalence of underweight, normal, overweight/ overfat and obese/excessive fat in school children was 3.4%, 39.7%, 33.0% and 24.0% versus 6.9%, 84.2%, 5.6% and 3.3% by body composition using the deuterium dilution technique and BMI-for-Age Z scores respectively.

**Conclusions:** It is apparent that the WHO recommended BMI-for-age to classify overweight and obesity underestimates the prevalence of overweight and obesity based on measures of body fatness. This needs to be confirmed in a larger study. Evidence-informed interventions should be based on more accurate estimates of the prevalence of overweight and obesity than can be provided by BMI.

**Keywords:** Obesity, nutrition assessment, anthropometry, body composition.

NUTRITION-FOCUSED PROGRAM IMPROVES READMISSION RATES OF MALNOURISHED HOSPITALIZED PATIENTS REGARDLESS OF DISCHARGE DISPOSITION

(1)Sulo, Suela Sulo; (2)VanDerBosch, Gretchen; (3)Summerfelt, Wm. Thomas; (4)Partridge, Jamie; (5)Hegazi, Refaat; (6)Sriram, Krishnan.

(1)PhD. MSc. Research & Development. Abbott Nutrition. Columbus. USA; (2)RD. Research & Innovation. Advocate Health Care. Downers Grove. IL. USA.; (3)PhD. Research & Innovation. Advocate Health Care. Downers Grove. IL. USA.; (4)PhD. MBA. Research & Development. Abbott Nutrition. Columbus. OH. USA.; (5)MD. PhD. Research & Development. Abbott Nutrition. Columbus. OH. USA.; (6)MD. Research & Innovation. Advocate Health Care. Downers Grove. IL. USA.

**Background and objectives:** Well defined discharge planning processes are key in limiting unwarranted hospital readmissions. Malnourished hospitalized patients are more likely than their well-nourished counterparts to experience readmissions if their nutritional needs are not appropriately addressed while in the hospital. We aimed to assess the impact of a hospital-based, comprehensive nutrition-focused quality improvement program (QIP) on readmission rates of malnourished patients discharged home with and without home health care services.

**Methods:** Data from 2588 patients (1269 QIP patients enrolled October 2014-April 2015, and 1319 retrospective historical controls admitted in the QIP hospitals October 2013-April 2014) were categorized into three discharge disposition categories including home without home health care (1266/48.9%), home with home health care (1043/40.3%), and other health care facility (e.g., inpatient rehabilitation, skilled nursing facility) (279/10.8%) patient subgroups. No patients going to other health care facilities were included in the pre-QIP cohort; therefore, no comparative analyses were pursued for this subgroup. QIP patients were admitted in four hospitals where electronic medical record (EMR) was upgraded to include Malnutrition Screening Tool (MST), and condition-specific oral nutritional supplements (ONS) were administered to all patients at risk for malnutrition (MST ≥ 2). In addition, nutrition education was provided to patients and caregivers during the inpatient stay and at discharge.

**Results:** Pre-QIP historical controls readmission rates were 22.2% (141/635) for patients discharged home without home health care, and 21.9% (150/684) for patients discharged home with home health care. Post-QIP readmission rates were 14.7% (93/631) and 18.7% (67/359) for the two patient groups; thus resulting in statistically significant relative risk reductions of 33.7% and 14.6% (p values < 0.05), respectively. These results were observed regardless of hospital length of stay, which was significantly reduced for both groups.

**Conclusions:** Unplanned 30-day readmissions among malnourished hospitalized patients discharged to home with and without home health care services can be significantly decreased through a nutrition-focused QIP that aims to screen, promptly administer ONS, and educate patients throughout their hospital stay and at dis-
charge. These results highlight the importance of discharge planning that emphasizes nutrition education for patients and caregivers, and continued ONS consumption post hospital discharge.

**Keywords:** Nutrition supplements, readmissions, discharge disposition, outcomes research/QI.

**Conflict of Interest Disclosure:** Financial support for the study was provided by Abbott Nutrition, Columbus, OH, USA.

Suela Sulo, Jamie Partridge, and Refaat Hegazi are Abbott Nutrition employees.

During the study conduct and analysis, Suela Sulo was an Advocate Health Care employee.

Dr Krishnan Sriram has received consultancy fees from Abbott Nutrition outside the present work.

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EFFECTS OF PHOTO-VOICE APPROACH ON 24-HOUR DIETARY RECALL ACCURACY AMONG UNIVERSITY OF IBADAN UNDERGRADUATE STUDENTS

(1)Ariyo, Oluwaseun; (2)Oloyede, Fiyinfoluwa.

(1)BSc. MSc. PhD. Nutritionist. Department of Human Nutrition. University of Ibadan. Nigeria; (2)BSc. Nutritionist. Department of Human Nutrition. University of Ibadan. Nigeria.

**Background and objectives:** The 24-hour recall is frequently used dietary assessment yet misreporting is common; therefore new approaches harnessing mobile technology are presently being explored. Photo-voice is an innovative qualitative method of participatory action research based on health promotion principles. However, it has not been used to its full potential in nutrition and dietetics. This study was designed to assess the effect of photo-voice approach on 24-hour dietary recall accuracy among undergraduate students.

**Methods:** This study was descriptive cross-sectional in design. Two hundred undergraduate students of University of Ibadan were selected using a simple random procedure from a pool of consenting students using WhatsApp-enabled smart phones. A semi-structured questionnaire was used to obtain information on socio-demographic characteristics and dietary intake. Respondents were prompted to capture and send every meal, snack and drink consumed daily and randomly selected for assessment over a 30-day period. Dietary intake was assessed using 24-hour multiple recall procedure, conducted using conventional approach and with the aid of photo-images. Nutrient intake was calculated using adapted Total Diet Assessment software and percentage difference in nutrient intake was calculated. Data were analysed using descriptive statistics and Chi-square test at p=0.05.

**Results:** Age was 21.0±1.8years, 57.5% were males and 95.0% lived on-campus. Intake of energy (1701.2±795.3; 1684.4±774.4) Kcal; protein (50.6±29.1; 50.1±28.1)g; fat (42.1±31.4; 41.6±27.2) g; and cholesterol (198.8±174.0; 188.4±172.0)g were higher with photo-voice than conventional approach respectively. Intake of Vitamin A (2149.5±1747.4; 2712.5±1903.4)RE; thiamin (1.1±1.0; 1.9±1.0)mg; and niacin (12.0±9.7; 12.4±10.4)mg were lower with

conventional than photo-voice respectively. However, riboflavin (0.9±0.8; 0.8±0.6)mg; Vitamin B6 (0.9±0.7; 0.6±0.5)mg and B12 (1.7±1.1; 1.4±0.8)mcg were higher in conventional than photo-voice. Recall accuracy for intake of calorie (+15.2%; -8.2%); thiamin (+21.4%; -58.5%); riboflavin (+27.3%; -24.2%) and niacin (+17.6%; -12.3%) was significantly different in female and male respondents respectively. Also, recall accuracy for intake of folate (+27.8%; -12.2%); B12(-58.3%; +13.3%); Calcium(+11.3%; -17.9%); phosphorus(+11.9%; -9.3%); sodium(-8.2%; -10.9%); potassium(+20.4%; -17.3%); zinc(+10.9%;-2.6%); iron(+14.1%; -8.2%) and magnesium(+16.9%; -8.2%) was significantly different in female and male undergraduates.

**Conclusions:** Photo-assisted 24-hour recall shows misreporting of micronutrients intake masked in conventional 24-hour recall. Recall accuracy differ significantly by gender with over-reporting of nutrients intake among female students.

**Keywords:** Dietary assessment, innovative methodology, nutrient intake; over-reporting; underreporting.

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VALIDITY OF BIOELECTRICAL IMPEDANCE ANALYSIS IN PREDICTING ADIPOSITY AMONG 8-11 YEARS OLD SENEGALESE SCHOOL-AGED CHILDREN

(1)Diouf, Adama; (2)Diongue, Ousmane; (2)Mégné, Ndé; (3)IDOHOU-DOSSOU, Nicole; (2)Thiam, Mbeugué; (4)Sembéne, Pape Malick;

(3)Wade, Salimata.

(1)PhD. Nutritionist. Laboratoire de Nutrition. Faculté des Sciences et Techniques. University Cheikh Anta Diop de Dakar. Senegal;

(2)Ms. Nutritionist. Laboratoire de Nutrition. Faculté des Sciences et Techniques. Université Cheikh Anta Diop (UCAD) de Dakar, BP 5005, Dakar-Fann. Senegal; (3)PhD. Nutritionist. Laboratoire de Nutrition. Faculté des Sciences et Techniques. Université Cheikh Anta Diop (UCAD) de Dakar, BP 5005, Dakar-Fann. Senegal; (4)MD. PhD. Physician. Division du Contrôle Médical Scolaire. Ministère de l'Education Nationale. Camp Jeremy Avenue Cheikh Anta Diop. BP 35 453. Dakar. Senegal.

**Background and objectives:** Childhood obesity is a serious public health challenge. Therefore accurate measurement of adiposity is essential to monitor the trend of child obesity in developing countries. This study aimed to develop and validate, for the first time, a bioelectrical impedance analysis (BIA) predictive equation for the assessment of adiposity among Senegalese school-aged children.

**Methods:** The study involved 151 pupils, 8-11 years old (74 boys, 77 girls) randomly selected from 4 elementary public schools in urban area of Dakar (Senegal). Weight and height were measured. Body composition: total body water (TBW), fat free mass (FFM) and percentage body fat (%BF) were determined by deuterium dilution method (DDM) used as the reference method. BIA was measured using multi-frequency analyzer and impedance index (height²/Z50 kHz) calculated. Stepwise backward multiple lin-
ear regression model was performed to develop the %BF equation in a subsample then validated it in the rest of the sample. Bland and Altman approach was used to assess the concordance between the two methods (bias and limits of agreement).

**Results:** Mean age (9.6±1.0 years) was comparable between boys and girls. FFM was higher in boys (24.6±6.9 kg) compared to girls (21.2±3.3 kg; P<0.001) and %BF lower in boys (5.1±4.6 kg) than in girls (5.3±3.8 kg; P<0.05). Overall, 11.3% of the children were obese (%BF ≥25% in boys, and ≥30% in girls). With BMI-z score, the prevalence of overweight/obesity was 4.6% (BMI>1). The equation newly developed for the prediction of adiposity was: %BF= -1.10*height²/Z50+3.14*sex+1.57*weight-4.347 (RMSE=4.5%, pure error=5.5%). The %BF estimated with the equation was highly and significantly correlated with the reference method (R²=0.76; P<0.001). Bland-Altman analysis showed non-significant mean bias (0.19±5.6% fat; P=0.764) and limits of agreement ranged from -10.7 to 11.1%. When the equation was applied to the whole sample, there were non-significant difference between %BF measured by DDM (18.7%±9.7) and that predicted by BIA (18.2%±7.5).

**Conclusions:** The %BF equation developed in the present study could be used as an alternative and accurate tool for the measurement of adiposity among school-aged children. Further cross-validation studies in a larger independent sample of African pupils is suitable.

**Keywords:** Adiposity, BIA, deuterium, school-aged children, Senegal.

**Further collaborators.**

Supported by International Atomic Energy Agency (IAEA) through the RAF/6/042 project and the Ministry of Higher Education and Scientific Research of Senegal (PAPES: Projet d'Appui aux Enseignantes-Chercheures du Sénégal).

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PERCEPTION OF PRINTED AND DIGITIZED PHOTOS IN THE QUANTIFICATION OF THE GLOBODIET-BRAZIL FOOD PORTIONS – RESULTS FROM THE VALIDA STUDY

**Background and objectives:** This validation study is a major component of an international 24-hour dietary methodology

adapted for Latin-American countries, starting with Brazil. The objective was to evaluate the perception of food portion quantification using printed and digitized photos.

**Methods:** The study was developed in Curitiba/PR (Brazil) with 58 adults. A total of 20 foods were evaluated from the GloboDiet Brazilian Album (n=96 foods). Each food had a series with up to six portion sizes. Three photos of each food were selected, representing small, medium and large portion sizes. Participants visited the study site in two moments, with a minimum of one-week interval, to evaluate the same food for both printed and digitalized photos (either in computer screen or tablet). Each participant evaluated on average 12 foods per session, randomly allocated, and had up to two minutes to choose the picture that best represented each pre-determined portion presented to them, which could be: exactly as the original photo, differing in size or in format. The difference between the correct and chosen portion (i.e., error) was analyzed, as well as concordance in using the photos and Spearman correlations.

**Results:** The percentage of the mean error was -1.1% (DP=11,2) for printed photos and -3.6% (DP=14,3) for digitized ones, showing no statistically significant difference (p=0.27). The concordance in using the photos was 91% and 90% for the printed and digitized photos, respectively. The foods with the highest number of distal chosen portions were the popcorn and feijoada. The correlation between correct and the estimated portions varied from 0.68 (scrambled egg) to 0.95 (noodles and popcorn) for printed and from 0.51 (feijoada) to 0.97 (potato) for digitalized photos. Tablet evaluations presented the largest errors, especially in the group of people with less years of education (p<0,05) and when the portions were presented in a different format (p<0,001) than shown in the photo.

**Conclusions:** As conclusion, the perception of using food portion printed photos was, in general, satisfactory although some foods were not very well evaluated. Furthermore, the evaluations using tablet resulted in worse results. We hope such results will contribute to future developments of dietary methodologies in Latin America.

**Keywords:** Food portion, photos, perception, food consumption.

**Further collaborators.**

We acknowledge the contributions of the GloboDiet Brazilian partners in Rio de Janeiro (Rosangela A Pereira, Marina Araújo, Rosely Sichieri), Sergipe (Danielle Góes) and France (Viktoria Knaze and Genèvieve Nicolas).

<footnote>(1) Nichelle, Pryscila Gharib; (2) Fisberg, Regina; (3) Almeida, Claudia Choma Bettega; (4) Maurício, Angelica; (5) Garmus, Lenine Maria;</footnote>

<footnote>(5) Mendes, Vanessa Cardoso; (6) Slimani, Nadia; (3) Marchioni, Dirce;</footnote>

<footnote>(7) Camey, Suzi Alves; (8) Crispim, Sandra.</footnote>

<footnote>(1) Federal University of Paraná; (2) D.Sc. Nutritionist. Departament of Nutrition. Public Health School. University of São Paulo. Brazil;</footnote>

<footnote>(3) D.Sc. Nutritionist. Departament of Nutrition. Federal University of Paraná. Brazil.; (4) D.Sc. Nutritionist. Departament of Nutrition.</footnote>

<footnote>Federal University of Paraná. Brazil.; (5) Nutritionist. Departament of Nutrition. Federal University of Paraná. Brazil.; (6) Independent researcher. France.; (7) D.Sc. Statistician. Departament of Statistics.</footnote>

<footnote>Federal University of Rio Grande do Sul. Brazil.; (8) PhD. Nutritionist. Departament of Nutrition. Federal University of Paraná. Brazil.</footnote>
# DEVELOPMENT OF CHINESE DIETARY INDEX FOR PREVENTING NON-COMMUNICABLE CHRONIC DISEASES AND ITS RELATIONSHIP WITH UNDERLYING RISK OF MAJOR CHRONIC DISEASES

(1)Su, Xiaoming; (2)Duan, Ruonan; (3)Zhang, Yanbo; (3)Lu, Weijia;

(4)Yang, Mingzhe; (5)Xue, Hongmei; (6)Liu, Yan; (7)Cheng, Guo.

(1)B.Med. West China School of Public Health. Sichuan University. China.; (2)MSc. Clinical Nutritionist. Jincheng People's Hospital. China.; (3)BMed Candidate. West China School of Public Health. Sichuan University. China.; (4)MSc. Researcher. Science and Technology Center, By-Health Co.,Ltd. China.; (5)PhD candidate. West China School of Public Health. Sichuan University. China.; (6)MSc. Clinical Nutritionist. Chengdu First People's Hospital. China.; (7)PhD. Professor. West China School of Public Health. Sichuan University. China.

**Background and objectives:** To develop the Chinese Dietary Index for Preventing Non-communicable Chronic Diseases (CDI-NCD), to investigate the association between overall diet quality and underlying risk of major chronic diseases.

**Methods:** Based on the current Chinese Dietary Guidelines, Chinese Dietary Reference Intakes (DRIs) and literature on development of dietary indices and association between diet and non-communicable diseases, CDI-NCD consists of 18 components. Diet quality of 572 healthy adults aged 25-65 years from Chengdu was evaluated using the scoring system of CDI-NCD. Differences in indicators for underlying risk of obesity, diabetes, dyslipidemia and hypertension among participants with higher, moderate and lower diet quality were tested.

**Results:** The mean CDI-NCD score for our population was 103.5±15.1. The diet quality of females was higher than that of males (P=0.001). Participants with normal weight had better diet quality than those with excess weight (P=0.004). Participants with higher diet quality had the lowest values of body mass index, insulin, triglycerides, low density lipoprotein and highest value of high density lipoprotein.

**Conclusions:** The CDI-NCD could evaluate the overall diet quality among Chinese adults. High diet quality, indicated by a high CDI-NCD score, was associated with lower underlying risk of major chronic diseases like obesity and dyslipidemia.

**Keywords:** Nutrition, dietary assessment, chronic disease, diet, dietary index.

**Further collaborators.**

Supported by Sichuan University Training Program of Innovation and Entrepreneurship for Undergraduates (project 20140316), National Natural Science Foundation of China (81472976 and 81673158) and research grant from the National 1000 Young Talents Program.

# NUTRITIONAL STATUS OF BELOW 3 YEARS CHILDREN AND INFANT AND YOUNG CHILD FEEDING PRACTICES IN INDIA-FINDINGS OF NATIONAL NUTRITION MONITORING BUREAU (NNMB) SURVEY

(1)Meshram, Indrapal; (2)Balakrishna, Nagalla; (3)R, HariKumar; (4)K, Mallikharjun Rao; (1)Arlappa, Nimmathota; (5)A., Laxmaiah.

(1)MD (SPM), National Institute of Nutrition, Hyderabad; (2)PhD (Bio-statistics), National Institute of Nutrition, Hyderabad; (3)DPh (Public Health), National Institute of Nutrition, Hyderabad; (4)PhD (Anthropology), National Institute of Nutrition, Hyderabad; (5)PhD (Public Health), National Institute of Nutrition, Hyderabad.

**Background and objectives:** Undernutrition is an important public health problem in India in spite of several nutrition intervention programs in operation since last 4 decades. The study was undertaken to assess infant & young child feeding practices and nutritional status of under 3 year children in rural area in 10 stated of India

**Methods:** A community-based, cross-sectional study was carried out in 10 states of India, using multistage random sampling method. Nutritional assessment was done using WHO Child Growth standard. Conceptual hierarchical framework was used as a basis for controlling for the explanatory factors in multivariate analysis

**Results:** A total of 4038 children below 3 years of age were covered. The study revealed that only 36% infants received breast feeding within 1 hour, while 37% received within 1-3 hours of birth. About 50% infants were exclusively breast feed up to 6 months. About 61% children of 6-11 months received complementary feeding at 6-8 month, 19% received before 6 months, while 17% has not yet received complementary feeding.

The prevalence of underweight, stunting and wasting was 38%, 42% and 22% respectively among less than 3 year children. It was observed that the risk of undernutrition among <3 year children was significantly (p<0.01) higher among boys, among children belonging to Scheduled caste and scheduled tribe communities, children from low income groups, among children of illiterate mothers and children without the facility of sanitary latrine. Among infants, the risk of underweight was significantly (p<0.01) higher among children belonging to scheduled tribe communities, children from low income groups, among children of illiterate mothers' children without the facility of sanitary latrine and among low birth weight babies, while the risk of stunting was higher among children belonging to low income groups, among children of illiterate mothers and children without the facility of sanitary latrine and among low birth weight babies.

**Conclusions:** Undernutrition is still an important health problem and is associated with income, community, literacy of mother, birth weight and sanitary latrine. Improving literacy status of mother thus improving socioeconomic condition and maternal nutrition during pregnancy and also sanitation & hygiene may improve the nutritional status of children

**Keywords:** Conceptual framework, Infant & young child feeding practices, income, nutrition assessment, undernutrition.
REPORTED DIETARY INTAKE AND FOOD SOURCES OF ZINC, SELENIUM, AND VITAMINS A, E AND C IN THE SPANISH POPULATION: FINDINGS FROM THE ANIBES STUDY

**Background and objectives:** Zinc, selenium, and the vitamins A (retinol and carotenes), E and C, all have specific biological functions that are involved mainly in the antioxidant defence system that has important implications for the development of chronic diseases. We aimed to assess the reported intake of these six nutrients, as well as the food that contributes to their sources of intakes.

**Methods:** Data were obtained from the Spanish ANIBES (Anthropometry, Intake and Energy Balance in Spain: anthropometric data, macronutrients and micronutrients intake, practice of physical activity, socioeconomic data and lifestyles) study, n=2009 (9-75 years old). The analyses were performed in the whole population and also in the plausible reporters after a misreporting analysis according to EFSA protocol. A three-day food record was used to collect the data.

**Results:** Mean reported intake for the whole population of zinc was 8.1±0.1 mg/d, (2.3-27.3 mg/d), selenium 75±1 µg/d, (14-265 µg/d), vitamin A 668 µg RE/d (2-11017 µg RE/d), retinol 364 ± 18 µg/d (0-10881 µg/d), carotenes 1735 ± 35 µg/d (13-13962 µg/d), vitamin E 7.0 ± 0.1 mg α-TE/d (0.7-55.2 mg α-TE/d) and vitamin C 84.4±1.4 mg/d (5.0-802.7 mg/d). The main source intakes for zinc were meat and meat products, for selenium were cereals and grains, for vitamin E oils and fat and for vitamin A and C vegetables.

**Conclusions:** There is an important percentage of the Spanish ANIBES population not meeting the recommended intakes for zinc, vitamin A and E, a reasonable percentage of people not meeting the recommendations of Vitamin C and a low percentage of people not meeting the selenium recommendations.

**Keywords:** ANIBES study, trace elements, vitamins, misreporting, food intake

**Conflict of Interest Disclosure:** The ANIBES study was financially supported by a grant from Coca-Cola Iberia through an agreement with the Spanish Nutrition Foundation (FEN). The funding sponsors had no role in the design of the study, in the collection, analyses, or interpretation of the data; in the writing of the manuscript, and in the decision to publish the results. The authors declare no conflict of interest.

VITAMIN A AND IRON CONTENT IN COMMON COMPLEMENTARY AND WEANING FOODS FOR CHILDREN IN NORTH WESTERN TANZANIA AND CENTRAL UGANDA

**Background and objectives:** The prevalence of Vitamin A and iron deficiencies among preschool children are 43% and 58.6% In Tanzania and 39% and 49% in Uganda respectively. These values are way beyond the WHO acceptable levels. The purpose of this work was to establish the vitamin A and iron adequacy of the most common weaning and complementary foods given to children in North western Tanzania and Central Uganda.

**Methods:** A cross-sectional study where 425 households selected through multistage sampling techniques were interviewed on the foods given to preschool children. Follow up home visits to randomly selected households were done to confirm details on preparation procedures. Six most common meals given to children were identified. Ingredients were obtained from the sites and transported to Department of food science, Universität für Bodenkultur (BOKU) Vienna, Austria within 48hours. Within 24hours of arrival, the meals were prepared following procedures validated by community and stored under refrigeration (-18 degrees C) to await analyses (<14days). HPLC analysis and microwave digestion followed by a flame atomic absorption spectroscopy were then used in establishing the content of provitamin A carotenoids (precursors of vitamin A) and iron content respectively. All the extractions and analysis were carried out in triplicate.

**Results:** Findings indicated that the six most common foods were maize-base porridge, steamed-mashed banana served with beans, boiled banana served with beans; boiled banana served with groundnut sauce, stiff porridge (Ugali) served with beans and boiled cassava served with beans.Analysis showed no trace of ei-

<footnote>(1)Olza, Josune; (2)Aranceta, Javier; (3)González-Gross, Marcela;</footnote>

<footnote>(4)Ortega, Rosa M; (5)Serra-Majem, Lluis; (6)Varela, Gregorio; (7)Gil, Ángel.</footnote>

<footnote>(1)PhD. Nutritionist. Institute of Nutrition and Food Science. Department of Biochemistry and Molecular Biology II. Faculty of Pharmacy. University of Granada.; (2)Professor. Department of Preventive Medicine and Public Health. Pamplona. Spain;
(3)Professor. ImFINE Research Group. Department of Health and Human Performance, Technical University of Madrid. Madrid. Spain; (4)Professor. Department of Nutrition. Faculty of Pharmacy. Madrid Complutense University. Madrid. Spain; (5)Professor. Research Institute of Biomedical and Health Sciences. University of Las Palmas de Gran Canaria. Faculty of Health Science. Las Palmas de Gran Canaria. Spain; (6)Professor. Department of Pharmaceutical and Health Sciences. Faculty of Pharmacy. CEU San Pablo University. Spain; (7)Professor. Department of Biochemistry and Molecular Biology II. Institute of Nutrition and Food Sciences. University of Granada. Granada. Spain.</footnote>

<footnote>(1)Ekesa-Onyango, Beatrice; (2)Nabuuma, Deborah; (3)Pamminger, Linda; (4)Namsifu, Godson; (5)Kennedy, Gina.</footnote>

<footnote>(1)PhD. Associate scientist-Nutrition. Healthy diets for sustainable food systems. Bioversity International.Kampala. Uganda.; (2)Msc. Research Fellow-Nutrition. Healthy diets for sustainable food systems. Bioversity International. Kampala. Uganda.; (3)Msc. Department of Nutritional sciences. University of Vienna. Vienna. Austria.; (4)Msc. Department of Food Nutrition and Consumer Studies. Sokoine University of Agriculture. Morogoro. Tanzania; (5)PhD. Senior Scientist-Nutrition. Healthy diets for sustainable food systems. Bioversity International. Rome. Italy.</footnote>
ther vitamin A or iron was observed in the maize-based porridge. Some iron 2.28mg/100gfw and 1.18mg/100gfw were observed in the stiff porridge served with beans and small fish and boiled banana served with red kidney beans respectively. The banana-based foods showed some content of vitamin A ranging from 23 to 43 RAE ug/100gfw.

**Conclusions:** Considering the estimated average requirements of iron and vitamin A for children being 4.1mg/d and 210-275 RAE ug/day respectively, these foods are poor sources of these nutrients in their current form. There is therefore need to explore opportunities of either modifying the preparation methods and incorporating more nutritious and diverse ingredients for better nutrition content.

**Keywords:** Vitamin A, Iron, Complementary foods, Weaning foods

the profile of nutrient contribution from dry soups was consistent with the expected RDAs contribution from vegetables.

**Conclusions:** A notable amount of nutrients are retained in vegetables after pre-processing and dehydrating and dry vegetable soups can therefore be considered a relevant source of vegetables and nutrients.

**Keywords:** Vegetables, nutrient composition, dehydration, nutrient retention.

**Conflict of Interest Disclosure:** All authors are employees of Unilever, a manufacturer of Food and Beverages products (including soups)

**Further collaborators:** Karin H. van het Hof. PhD. Unilever R&D Vlaardingen. Vlaardingen. The Netherlands

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## NUTRITIONAL QUALITY OF DRIED VEGETABLES AND VEGETABLE SOUPS

(1)van het Hof, Karin; (2)Grün, Christian; (3)Basendowski, Silke; (4)Spraul, Martin; (5)van Buren, Leo; (3)Vollmer, Gabi; (2)Newson, Rachel.

(1)PhD. Nutritionist. Unilever R&D Vlaardingen. The Netherlands;

(2)PhD. Unilever R&D Vlaardingen. Vlaardingen. The Netherlands;

(3)Unilever R&D Heilbronn. Heilbronn. Germany; (4)PhD. Unilever R&D Heilbronn. Heilbronn. Germany; (5)Unilever R&D Vlaardingen. Vlaardingen. The Netherlands.

**Background and objectives:** Dehydration of food has been used for centuries as a technique to increase shelf life and preserve nutrient quality during storage. However, degradation or loss of nutrients may occur during the drying process, either due to applied heat or leaching due to water removal. Therefore, we investigated nutrient retention during pre-processing and drying of vegetables used to prepare dry vegetable soups in 2 studies.

**Methods:** First, nutrient levels were determined in fresh industrial tomato, onion, and lentil produces, as well as in the industrially dried products obtained from the same batches. Vitamin C and lycopene were selected as representatives for heat- and storage-labile nutrients in tomato. For onion, vitamin C and flavonols were taken, whereas folates were chosen as nutrient markers for lentils. Potassium contents were determined for all vegetables. Second, the overall nutritional quality of dry vegetable soups was assessed and compared with recommended dietary allowances (RDAs) and nutrition databases. Nutrients, including dietary fiber, minerals, vitamins, and carotenoids were determined in ten commercial dry vegetable soup varieties.

**Results:** During the pre-processing and dehydration of tomato, lycopene remained stable, whereas 70% of vitamin C was retained. With regard to onions, 29% of vitamin C and 51% of flavonols were retained. Losses of folates during processing of lentils was between 10-25%, depending on the process applied. Potassium levels remained largely unaltered for all vegetables, indicating limited losses attributable to leaching. The nutritional profile of dry vegetable soups aligned well with nutritional databases and

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## ANEMIA AND USE OF IRON SUPPLEMENTATION IN CHILDREN LIVING IN SOCIAL VULNERABLE CITIES IN THE SOUTH REGION OF BRAZIL

(1)Letícia Frizzi Silva, Débora; (2)Lang, Regina Maria Ferreira;

(3)Moreira, Julia Dubois; (4)Dallazen, Camila; (5)da Silva, Sara Araújo;

(5)Gonçalves, Vivian Siqueira Santos; (5)Nilson, Eduardo Augusto Fernandes; (6)Höfelmann, Doroteia; (4)Vitolo, Márcia Regina;

(6)Crispim, Sandra.

(1)Federal University of Paraná; (2)Nutritionist. Department of Nutrition. Federal University of Paraná. Curitiba, PR. Brazil;

(3)Nutritionist. Department of Nutrition. Federal University of Santa Catarina. Florianópolis, SC. Brazil.; (4)Nutritionist. Federal Foundation University of Health Sciences, Porto Alegre. Porto Alegre, RS. Brazil.; (5)Coordination of food and nutrition. Ministry of Health. Brasilia-DF. Brazil.; (6)Department of Nutrition. Federal University of Paraná. Curitiba-PR. Brazil.

**Background and objectives:** Anemia is one of the major nutritional deficiencies in children under five years old, with consequences for growth and development. In Brazil, the National Iron Supplementation Program (PNSF) was introduced in 2005, aiming at controlling and preventing anemia in children by administering prophylactic doses of sulfate ferrous to those of 6-24 months. The objective of this study was to evaluate the prevalence of anemia and its association with the iron supplementation among children aged 12-59 months residing in cities that are part of Brazilian plan without extreme poverty from South of Brazil.

**Methods:** The study comprised children from 48 municipalities. Blood sampling was collected by peripheral venous puncture, followed by measurement of hemoglobin (Hb) using HemoCue®. Weight and length/height measurements were measured and face-to-face interviews were performed, using a structured questionnaire to obtain complementary information (e.g., socioeconomic, demographic and supplement use). Anemia was diagnosed when levels of Hb<11g/dL. Children were grouped according to the administration of sulfate ferrous: "using/used" (n=688) and "never used" (n=708). Double data entry and validation was done using
Epi-Data®. To evaluate the statistical association, chi-square test was performed in SPSS version 22, considering p<0,05.

**Results:** A total of 1511 children participated in the study with levels of Hb. The mean Hb was 12.8±1.13 g/dL. The majority of the children were male (51.5%), aged ≥24 months (71.4%), had a monthly family income of ≤1 minimum wages (32.7%) and were being monitored in health services since birth (93.1%). From the total sample, 49.3% were never supplemented and the prevalence of anemia was 4.9% (n=74). While the prevalence for children aged 12-24 months was 9.9% (n=43), for those with more than 24 months was 2.9% (n=31). When we compared the prevalence of anemia between those that were supplemented and those who were not, the prevalence were 4,2% (n=30) and 5.5% (n=38), respectively, with no statistical difference (p=0.26).

**Conclusions:** We conclude that the prevalence of anemia is considered acceptable, according to the World Health Organization. Furthermore, the determinants that are associated to this prevalence deserves further investigation, which seems not to be related to iron supplementation.

**Keywords:** Anemia; Children; Brazil; Iron Supplementation, Healthy Policy.

**Further collaborators:** We acknowledge the contribution of Dr. Daniela Cardozo Tietzmann and David Gonzalez to this study.

**Results:** The Index score is not dependent on energetic value (r=0.24). There is a high correlation between the score assigned by experts and the DQI-DFG result (r=0.78 until r=0.97).

**Conclusions:** Consensus result performed among experts legitimates the concepts that justify DQI-DFG; there was an agreement between menu quality evaluation by experts and the results shown by DQI-DFG; the correlation between food groups scores were weak and Index application result by evaluators agreed with the final DQI-DFG. The DQI-DFG update satisfied validation and reliability criteria.

**Keywords:** Healthy eating. Index. Diet Quality Index. Food and Nutrition Education.

**Background and objectives:** Background. Inadequate eating habits represent one of the main factors associated with chronic non-communicable diseases and nutritional deficiencies; food guides provide guidelines aiming to contribute to the promotion of health. The Diet Quality Index associated with the Digital Food Guide (DQI-DFG) was developed in 2011 from guidelines of School of Public Health at Harvard University, with adjustments for the Brazilian food habits. Objective. Analyze, improve and validate the Diet Quality Index associated with the Digital Food Guide, as a tool for food intake and diet quality assessment.

**Methods:** Method. The following psychometric properties were used: (a) content validation by building a consensus on healthy eating and diet quality index among experts; (b) construct validation through enhancement of a reference diet to determine recommended serving sizes and consumption ranges; correlation between each food group score and energy value; comparative analyses between menu assessment by experts and DQI-DGF application results to confirm the score criterion; (c) reliability from agreement analyses between the result shown by the evaluators and result presented by DQI-DFG; correlation between each food group score and total score Index to understand the relationship among this variables.

**Background and objectives:** Anemia affects 1.65 billion individuals worldwide, especially young children and women of reproductive age. Anemia prevalence in pregnant women (hemoglobin <110 g/L) in Malaysia is estimated to be 38%. However, there is limited evidence on the prevalence of anemia and

iron deficiency anemia (IDA) in non-pregnant Malaysian women and no studies have adjusted ferritin values for inflammation. This cross-sectional study aims to determine the prevalence of anemia and IDA in a sample of healthy non-pregnant Malay and Chinese women (19-45 years) living in Kuala Lumpur, Malaysia.

**Methods:** Fasting blood samples were collected from women living in Kuala Lumpur, Malaysia (n=210) and, for comparison purposes, from women living in Vancouver, Canada (n=206). Malaysian participants were of Malay (n=105) and Chinese (n=105) descent; Canadian participants were of European (n=110) and Chinese (n=96) descent. Samples were analyzed for hemoglobin, plasma vitamin B12, serum retinol binding protein (RBP), serum iron biomarkers (soluble transferrin receptor [sTfR] and ferritin),
and acute and chronic inflammation biomarkers (C-reactive protein [CRP] and α-1 acid glycoprotein [AGP]).

**Results:** Anemia (hemoglobin <120 g/L) prevalence was higher in Malaysian women compared to Canadian women (18% and 7%, respectively; p=0.001). In Malaysia, 11% (n=24) of women had IDA (based on hemoglobin <120 g/L and either inflammation-adjusted ferritin <15 µg/L or sTfR >8.3 mg/L), compared to 3% (n=6) of Canadian women. No significant ethnic differences were found among women in anemia or IDA prevalence. Depleted iron stores (inflammation-adjusted ferritin <15 µg/L) were found in 21% of Malaysian women and in 15% of Canadian women. The prevalence of acute (CRP >5 mg/L) and chronic (AGP >1 g/L) inflammation was 7% and <1%, respectively, in Malaysian participants, compared to 4% and <1% in Canadian participants. Adjustment for inflammation (using AGP and CRP biomarkers) did not change the prevalence of IDA in Malaysian or Canadian women. There was no biochemical evidence of vitamin A deficiency (RBP <0.7 µmol/L) and <1% of women had vitamin B12 deficiency (B12 <148 pmol/L) in both populations.

**Conclusions:** These findings suggest that over half the anemia in Malaysian non-pregnant women is attributed to iron deficiency. Further research is required to investigate the prevalence of IDA in a nationally representative sample.

**Keywords:** anemia, hemoglobin, iron, Malaysia, women

60 to 72 y, a protein intake of at least 1.2 g/kg bw.d showed an improved muscle strength and better mobility compared with participants consuming not more than 0.8 g/kg bw.d. A protein consumption of 1.2-1.6 g/kg bw.d was associated with an increased lean body mass compared with 0.8-1.0 g/kg bw.d in women aged 68 y (median). In postmenopausal women (75 y, median), a protein intake of 81 g/d was associated with a higher bone-free lean body mass and body cell mass than in those participants only consuming <66 g/d. Two recent prospective cohort studies showed within the 3y observational period a lower incidence of frailty in the quartile with the highest protein intake (1.27 g/kg bw.d and 1.2 g/kg bw.d, respectively) compared to the quintile with lowest intake (<1.0 g/kg bw.d)

**Conclusions:** The results of our systematic evaluation of actual research work confirms our working hypothesis that a protein intake >0.8 g/kg bw.d can slow down impairments in protein/amino acid metabolism during aging. The working group, thus, recommend to increase the RDA for older adults (>65 y) for protein up to 1.0 g/kg bw.d.

**Keywords:** Protein intake, dietary allowances, elderly, systematic review.

**Further collaborators.**

Working group of the German, Austrian and Swiss Nutrition Societies (D-A-CH)

144/1520

NEW MIDDLE EUROPEAN REFERENCE VALUES FOR DIETARY PROTEIN INTAKE IN THE ELDERLY

(¹)Stehle, Peter; (²)Oberritter, Helmut; (³)Richter, Margrit.

(¹)PhD. Prof. Nutritionist. Institute for Nutrition and Food Sciences. University of Bonn. Germany; (²)PhD. Nutritionist. German Nutrition Society. Bonn. Germany; (³)PhD. Nutritionist. German Nutrition Society. Bonn. Germany.

**Background and objectives:** Physiological aging is characterized by a steadily reduced protein synthesis capacity, a decreased postprandial amino acid availability and a diminished peripheral blood flow. We hypothesize that these age-dependent impairments in protein/amino acid metabolism can be slowed down by a nutritive protein intake >0.8 g/kg bw.d as recommended for younger adults.

**Methods:** In the obligatory updating of the D-A-CH Reference Values (RDAs) for Nutrient Intake we conducted a systematic literature review (PubMed, Embase; last entry: December 2016) identifying observational and clinical studies. The selection of and data extraction from studies included in the review were performed by two independent reviewers. The Working Group Of the German, Austrian and Swiss Societies evaluated the results obtained and draw conclusions with respect to the revision of RDAs for older adults.

**Results:** In several prospective cohort studies and a single cross-sectional trial, a positive correlation between protein intake and functionality was observed in older adults. In women aged

144/1563

IMPACT OF MICRONUTRIENT POWDERS COMBINED WITH MALARIA CHEMOPREVENTION ON ANEMIA, MALARIA AND COGNITIVE DEVELOPMENT: A CLUSTER-RANDOMIZED STUDY IN MAIAN CHILDREN

(¹)Diarra, Niele Hawa; (²)Roschnik, Natalie; (³)Clarke, Sian; (⁴)Sacko, Moussa; (⁵)Dicko, Yahia; (⁶)Verhoef, Hans; (⁷)Saye, Renion; (⁸)Lal, Sham; (⁹)Pisani, Lauren; (¹⁰)Boivin, Michael.

(¹)MD. School Health and Nutrition Coordinator. Save the Children International. Mali; (²)MSc. Senior Nutrition Advisor. Save the Children. UK; (³)PhD. Senior Lecturer. London School of Hygiene and Tropical Medicine; (⁴)PhD. Senior Parasitologist. Institut National de Recherche en Sante Publique. Mali; (⁵)MD. Adolescent Sexual and Reproductive Health Coordinator. Save the Children. Mali; (⁶)PhD. Senior Lecturer. MRC International Nutrition Group. London School of Hygiene and Tropical Medicine. UK/ Division of Human Nutrition and Cell Biology Group. Wageningen University. The Netherlands; (⁷)MSc. PhD student. Institut National de Recherche en Sante Publique. Mali; (⁸)MSc. Research Fellow. London School of Hygiene and Tropical Medicine. UK; (⁹)MSc. Senior Research Specialist. Save the Children. USA; (¹⁰)PhD. Professor. Michigan State University. USA.

**Background and objectives:** Anaemia is common in pre-school African children, and is mostly due to iron deficiency and Plasmodium infection. Combining iron interventions with interventions against malaria may be more efficacious and safer than
iron interventions alone. We examined the impact of home fortification with micronutrient powders, delivered alongside seasonal malaria chemoprevention, parenting education and community based preschools in rural communities in Southern Mali over a two year period.

**Methods:** Sixty communities with community preschools were randomly allocated to the intervention and control group. All 60 study communities received two rounds of seasonal malaria chemoprevention during the peak malaria season (Oct-Dec), targeting all children aged 3-59 months. The 30 intervention communities then received daily home fortification with micronutrient powders containing 12,5 mg of elemental iron for four months (Jan-April) targeting all children aged 6-59 months. Two cross sectional surveys were conducted at baseline (May-June 2014) and endline (May-June 2016) to compare Plasmodium infection, nutritional indices and cognitive performance in children aged 3 and 5 years (N=1,105 and N=1,033 respectively).

**Results:** At baseline in 2014, 63% and 53% of children aged 3y and 5y were anaemic, 49% and 60% had Plasmodium infection and 43% and 34% were stunted. Despite high reported coverage for both chemoprevention and home fortification, the preliminary results from June 2016 found that after three years of intervention there was little discernible difference in anaemia (<110g/L) between the two intervention groups; 51.8% vs 48.9% in children aged 3 years, and 46.6% vs 40.6% in children aged 5 years; and negligible effects for stunting. The full results of the 2016 survey will be presented at ICN meeting.

**Conclusions:** Home fortification with iron-containing micronutrient powders is recommended by WHO where the prevalence of anaemia is over 20%. This study found that micronutrient supplementation, when combined with seasonal chemoprevention against malaria, had limited impact on childhood anaemia in an area of high prevalence, despite good uptake of both interventions.

**Keywords:** Micronutrient, anaemia, malaria, home fortification, iron.

**Further collaborators.**

Seybou Diarra, Philippe Thera, Kalifa Sidibe, Modibo Bamadio, Yvonne Griffiths, Mouctar Coulibaly, Fatoumata Dougnon, Bore Saran Diakité, Bonaventure Maiga

**Background and objectives:** Thyroidal iodine uptake and turnover has been measured in adults using radioactive iodine tracers, but these cannot be used safely in women of reproductive age, pregnant women or children. Iodine 129 (129I) is a semi-stable isotope that is considered safe for use in human trials. Our objective was to assess 129I as a novel tracer for the assessment of fractional absorption (129IFA) and thyroidal uptake (129ITU).

**Methods:** We administered an oral physiological dose of 129I (129I Dose) to euthyroid young adults with adequate iodine intake. We collected baseline complete urines, complete feces and spot plasma samples. After tracer consumption, we collected complete urines (over seven days), complete feces (three days), and spot plasma (five days, with eight withdrawals on first day). The subjects followed an iodine-standardized diet from three days before until three days after tracer consumption. We measured 129I in alkaline-extracted plasma (129IP), urine (129IU) and feces (129IF) samples by a multicollector ICP-MS by use of isotope dilution analysis with 127I. 129IFA is evaluated as 129I Dose minus 129IF; 129ITU is evaluated as129IFA minus 129IU; 129IP kinetics allows the evaluation of thyroid uptake patterns.

**Results:** Four male and four female euthyroid (mean±SD TSH 1.3±0.5 mU/L; T4 110.0±19.7 nmol/L) participants with a median (IQR) urinary iodine concentration of 118 (101-140) µg/L consumed 12.42±0.05 µg 129I. 129IU was first detected ≈1 hour after administration and a median cumulative 129IU of 6.9 (6.4-7.4) µg was excreted within 48 hours; this corresponds to 54.8±10.6 % of the 129I Dose. **Results:** **Conclusions:** A novel 129I-tracer based method successfully quantified 129I renal excretion patterns, showing that ≈55% of ingested oral iodine is excreted in the subsequent 48 hours. Quantification of 129IFA and 129ITU thyroidal uptake patterns are pending. This promising and safe tracer methodology can safely be used in all age groups to investigate a variety of research questions on thyroid health and iodine nutrition, including iodine requirements in pregnancy, lactation and infancy.

**Keywords:** 129I, mass-spectroscopy, dietary iodine, thyroid uptake, iodine kinetic.

<footnote>(1)Galetti, Valeria; (2)van der Reijden, Olivia; (3)Mannhart, Andrea;</footnote>

<footnote>(4)Krzystek, Adam; (4)Zeder, Christophe; (5)Andersson, Maria; (5)Herter-Aeberli, Isabelle; (6)Zimmermann, Michael Bruce.</footnote>

<footnote>(1)PhD. Laboratory of Human Nutrition. Institute of Food, Nutrition and Health. ETH Zurich. Switzerland; (2)MSc. Laboratory of Human Nutrition. Institute of Food, Nutrition and Health. ETH Zurich. Switzerland.; (3)BSc. Laboratory of Human Nutrition. Institute of Food, Nutrition and Health. ETH Zurich. Switzerland.;</footnote>

<footnote>(4)Laboratory of Human Nutrition. Institute of Food, Nutrition and Health. ETH Zurich. Switzerland.; (5)PhD. Laboratory of Human Nutrition. Institute of Food, Nutrition and Health. ETH Zurich. Switzerland.; (6)MD. Laboratory of Human Nutrition. Institute of Food, Nutrition and Health. ETH Zurich. Switzerland.</footnote>
# INTERRELATIONSHIPS BETWEEN FATTY ACID COMPOSITION IN PLASMA CHOLESTEROL ESTERS AND PHOSPHOLIPIDS IN MEN AND WOMEN - A POOLED ANALYSIS

(1)Marklund, Matti; (2)Pingel, Ronnie; (1)Rosqvist, Fredrik; (3)Lindroos, Anna Karin; (4)Eriksson, Jan; (5)Vessby, Bengt; (6)Oscarsson, Jan; (7)Lind, Lars; (8)Risérus, Ulf.

(1)PhD. Department of Public Health and Caring Sciences, Clinical Nutrition and Metabolism. Uppsala University. Sweden.; (2)PhD. Department of Public Health and Caring Sciences. Uppsala University. Sweden.; (3)PhD. National Food Agency. Sweden. Department of Internal Medicine and Clinical Nutrition. University of Gothenburg. Sweden; (4)MD, PhD. Department of Medical Sciences, Clinical Diabetes and Metabolism. Uppsala University. Sweden.; (5)MD, PhD. Department of Public Health and Caring Sciences, Clinical Nutrition and Metabolism. Uppsala University. Sweden.; (6)MD, PhD. AstraZeneca Gothenburg. Sweden.; (7)MD, PhD. Department of Medical Sciences, Cardiovascular Epidemiology. Uppsala University. Sweden; (8)MMED, PhD. Department of Public Health and Caring Sciences, Clinical Nutrition and Metabolism. Uppsala University. Sweden.

**Background and objectives:** Fatty acid (FA) proportions in cholesterol esters (CE) and phospholipids (PL) are widely used as biomarkers of dietary fat quality. Information on how proportions in these fractions correlate could have implications for interpretation and use of FA biomarkers in observational and interventional studies. The objectives of this study were to investigate correlations between FA proportions in CE and PL among free-living individuals and to assess how diet-induced alterations of FA proportions correlate between fractions.

**Methods:** Spearman's rank correlation coefficients (rs) between FA proportions (% of total FA) in circulating CE and PL were calculated separately in 8 individual study populations (total n=2052) and pooled by inverse-variance weighted meta-analysis. Additionally, each study population was stratified by age, sex, BMI, and type 2 diabetes status and strata-specific rs were pooled by meta-analysis. Furthermore, rs between FA changes in CE and PL during isocaloric dietary interventions were calculated separately per treatment in two randomized controlled trials (total n=79).

**Results:** Overall, FA proportions in CE and PL ranked individuals similarly. Polyunsaturated FA proportions correlated especially strongly between fractions, with pooled rs (95% CI) ranging from 0.74 (0.72; 0.76) for α-linolenic acid to 0.92 (0.91; 0.93) for eicosapentaenoic acid. Weak correlations (pooled rs< 0.4) were only observed for, palmitic acid and stearic acid, with pooled rs (95% CI): 0.29 (0.24; 0.33) and 0.30 (0.26; 0.34), respectively. Correlations were not evidently affected by age, sex, BMI, or type 2 diabetes. Strong correlations (rs≥ 0.6) between diet-induced FA changes in CE and PL were observed for most PUFA and especially eicosapentaenoic acid, where rs ranged 0.85-0.94 after different treatments.

**Conclusions:** Correlations between FA proportions in CE and PL were generally strong, suggesting that FA proportions in these fractions can be used interchangeably in diverse populations.

However, caution is advised when comparing results from studies assessing palmitic acid or stearic acid in different lipid fractions.

**Keywords:** Biomarker; Fatty acid; Cholesterol ester; Phospholipid; Meta-analysis.

# ZINC INTAKES AND DIETARY SOURCES IN AN IRISH ADULT POPULATION

(1)Lenighan, Yvonne; (2)Nugent, Anne P.; (3)Walton, Janette; (4)Flynn, Albert; (5)Roche, Helen; (6)McNulty, Breige A.

(1)Research Nutritionist. UCD Conway Institute and UCD Institute of Health. University College Dublin. Ireland.; (2)Lecturer in Nutrition. UCD Institute of Food and Health. University College Dublin. Ireland; (3)Research Fellow. School of Food and Nutritional Sciences. University College Cork. Cork. Ireland.; (4)Professor in Nutrition. School of Food and Nutritional Sciences. University College Cork. Cork. Ireland.; (5)Professor in Nutrition. UCD Conway Institute and UCD Institute of Food and Health. University College Dublin. Dublin. Ireland.; (6)Lecturer in Nutrition. UCD Institute of Food and Health. University College Dublin. Dublin. Ireland.

**Background and objectives:** Adequate zinc intakes are important for growth, immune function and cognitive health. Zinc deficiency affects approximately 17% of the global population, with young children and older adults at increased risk. There is currently limited data on zinc intakes in the Irish population. Therefore the aim of this analysis is to determine zinc intakes and sources in Irish adults using data from the cross-sectional National Adult Nutrition Survey (NANS).

**Methods:** The NANS assessed habitual food and beverage intake between 2008 and 2010 for 1500 Irish adults (18-90 years) using a 4-day semi-weighed food diary. 2552 food codes were aggregated into 33 food groups, representative of the overall diet. Differences in zinc intakes were assessed using one-way ANOVA and independent t-tests.

**Results:** Mean daily zinc intakes were 10.3mg in the total population. Males (11.6mg) presented greater intakes than females (9.0mg) (P<0.001). In comparison to the younger males (12.4mg), intakes of zinc were significantly lower in older males (10.2mg) (P=0.004). Smokers presented significantly lower zinc intakes than non-smokers in the total population, while intakes were significantly greater in supplement users than non-supplement users (P<0.05). Similar differences were observed when the population was split by gender (P<0.05). The Institute of Medicine recommend daily zinc intakes of 11mg and 8mg in males and females, respectively. These intakes were achieved by the current population. The dominant food groups contributing to zinc intakes were unprocessed red meat (22.3%), wholemeal/brown bread and rolls (10.1%), processed red meat (7.5%), white meat (5.7%) and white bread, rolls, scones and croissants (5.1%).

**Conclusions:** In summary, zinc intakes are adequate in this Irish population; however, they should be monitored continuous-
ly, particularly in older men as they presented significantly lower
intakes.

**Keywords:** Zinc, dietary intakes, dietary sources.

144/1740

PHYTIC ACID CONTENT DETERMINES THE IRON
BIOAVAILABILITY FROM WHEAT GENOTYPES

(1)Jyrwa, Yvette Wilda; (2)Palika, Ravindranadh; (3)R., Ananthan;

(4)Pullakhandam, Raghu; (5)Longvah, Thingnganing.

(1)M.Sc. Micronutrient Research Division. National Institute of Nutrition. Indian Council of Medical Research. Hyderabad. India;

(2)M.Sc. Micronutrient Research Division. National Institute of Nutrition. Indian Council of Medical Research. Hyderabad. India.;

(3)PhD.Food Chemistry Division. National Institute of Nutrition.
Indian Council of Medical Research. India. Hyderabad.; (4)Phd.
Micronutrient Research Division. National Institute of Nutrition.

Indian Council of Medical Research. Hyderabad. India.; (5)Director
In- Charge. National Institute of Nutrition. Indian Council of
Medical Research. Hyderabad. India.

**Background and objectives:** Biofortification envisages iden-
tification, selection and stabilization of genotypes with higher nu-
trient contents. However ensuring bioavailability of nutrients is
required for the intended beneficial outcome in target population.
The objective of the present study is to understand the relation-
ships amongst iron content, iron bioavailability and phytic acid
content among wheat genotypes.

**Methods:** Iron and phytic acid content in 22 different wheat genotypes were estimated using standard methods. The iron dialyzability after simulated digestion of wheat flour was performed in a 6-well plate format. The iron content in the digesta and dialysable fraction were estimated by atomic absorption spectrometry.

**Results:** The iron and phytic acid content of wheat genotypes varied from 3.3 to 4.9mg/100g and 2.2 to 8.9mg/100g dry weight, respectively. The dialyzable iron and percent dialyzability are in the range 0.24 to 0.72mg/100g and 7.4 to 16%, respectively among the 22 wheat genotypes tested. There was no significant correlation amongst iron content with that of either iron dialyzability or phytic acid content. However, phytic acid content negatively correlated with that of dialyzable iron content (correlation coefficient-0.0641 p=0.0013)

**Conclusions:** The above results suggest significant variability in iron, phytic acid content and bioavailable iron among wheat genotypes. The phytic acid but not total iron appears to determine the extent of iron dialyzability. These results suggest that iron and phytic acid content alone appear to provide the key selection marker for breeding wheat to improve iron status

**Keywords:** Biofortification, bioavailability, in vitro digestion, phytic acid.

144/1755

MEASUREMENT OF HUMAN MILK INTAKE USING STABLE ISOTOPE IN INDIAN RURAL INFANTS FROM BIRTH TO 24 MONTHS: A LONGITUDINAL STUDY

(1)Deshmukh, Urmila; (2)Thomas, Tinku; (3)Swaminathan, Sumathi;

(4)Kurpad, Anura.

(1)MD. Pediatrician. Division of Nutrition. St John's Research Institute. St. John's National Academy of Health Sciences.
Bangalore. India.; (2)PhD. Biostatistician. St John's Research Institute. St. John's National Academy of Health Sciences.

Bangalore. India.; (3)PhD. Nutritionist. St John's Research Institute. St. John's National Academy of Health Sciences.
Bangalore. India.; (4)MD. Physiologist. Division of Nutrition. St John's Research Institute. St. John's National Academy of Health Sciences. Bangalore. India.

**Background and objectives:** Exclusive breastfeeding for first six months of life, followed by complementary feeding and continuation of breastfeeding is advocated by international health agencies. In this study, human milk intake from birth to 24 months was measured, and its associations with maternal and infant factors were studied.

**Methods:** In this community based, longitudinal study data collected were: Socio-demographics, anthropometry (weight, length and skin-folds of mother and offspring), morbidity and diet recall. Using deuterium oxide (30g) dose-to-mother technique, saliva samples of mother and infant were collected (on day 0, 1, 2, 3, 4, 13, and 14) at 1, 3, 6, 12, 18 and 24 months post-partum, and were analyzed by Fourier transform infrared spectroscopy. The human milk intake and non-milk oral water intake were estimated using a standardized method. Associations between human milk intake and maternal and infant factors were studied using correlations and linear mixed models. Approval from the Institutional Ethics Committee was obtained.

**Results:** Thirty rural mother-offspring pairs (age 29±3y and 30±7d, respectively) were enrolled. The mean human milk intake increased from 1st to 6 months from 686 ±162 to 844 ±117 g/d (p<0.001), and then decreased to 534 ±239 and 380 ±239 g/d at 18 and 24 months respectively. Rate of breastfeeding fell from 100% at one month to 97% at 6, 78% at 12 and 52% at 24 months. The human milk intake in first six months was positively associated with maternal fat mass, infant weight, length, sum of skin-folds and anthropometric z scores. After controlling for parity, birth weight, age and gender, human milk intake was associated with only infant's weight (p<0.001).

**Conclusions:** The HMI increased from first to sixth month, and then decreased significantly after 12 months till 24 months of age in infants who continued to be breastfed. High rate of breastfeeding beyond six months underpins need to assure adequacy of maternal nutrient stores, in this population.

Funding agency: The International Atomic Energy Agency, Vi-
enna, Austria.

**Keywords:** Human milk intake, breastfeeding, infants, India.
## VALIDATION OF THE DIETARY REFERENCE INTAKES FOR PREDICTING ENERGY REQUIREMENTS IN ELEMENTARY SCHOOL-AGE CHILDREN

(1)Kim, Eun-Kyung; (2)Ndahimana, Didace; (3)Kim, Ye-Jin; (4)Park, Jonghoon; (5)Ishikawa-Takata, Kazuko.

(1)PhD. Department of Food and Nutrition. Gangneung-Wonju National University. South Korea.; (2)PhD student. Department of Food and Nutrition. Gangneung-Wonju National University. South Korea.; (3)MSc student. Department of Food and Nutrition. Gangneung-Wonju National University. South Korea.; (4)PhD. Professor. Department of Physical Education. Korea University. South Korea.; (5)PhD. Head of Section of Nutritional Care and Management. Department of Nutritional Education. National Institute of Health and Nutrition. Japan.

**Background and objectives:** Accurate assessment of children energy requirements is important for the prevention of pediatric obesity, which has been a big public health challenge in recent years. The purpose of the present study was to examine the accuracy of Dietary Reference Intakes for predicting energy requirements in elementary school-age children, by using the doubly labeled water as a reference method.

**Methods:** The doubly labeled water (DLW) method was used to measure total energy expenditure (TEE) in 25 elementary school children aged 9-11 years. The accuracy of DRI predictive equations for TEE was assessed by comparing the predicted TEE (TEEDRI) to the measured TEE (TEEDLW), based on the mean percentage error (bias), the Root Mean Squared prediction Error (RMSE) and the percentage subjects predicted within 10% of TEEDLW. The Bland-Altman plot was used to assess agreement between the two methods.

**Results:** The DRI predictive equation overestimated TEE by 242.7 kcal/ day (12.6%) in boys, and underestimated TEE by 31.2 kcal/ day (-1.6%) in girls, with the RMSE of 376.7 and 216.8 kcal/ day, respectively. The percentage of accurate predictions was 28.6% in boys and 63.6% in girls. The bland-Altman plot showed larger limits of agreement in boys compared to girls. In addition, there was a proportional bias in case of boys, with a tendency of underprediction in subjects with low TEE and overprediction in subjects with high TEE. On the contrary, no proportional bias was observed in case of girls.

**Conclusions:** These results suggest that in elementary school-age boys, the DRI predictive equation could be inaccurate in predicting energy requirements. In girls, the present study indicates that the DRI equation could be used accurately at the group level. However, caution should be taken when estimating energy requirements for individual girls as this equation may result in inaccurate estimation in a large percentage of subjects. More studies are required to confirm our findings, and to assess the accuracy of the DRI equations in other groups including obese children, younger children and adolescents.

**Keywords:** Children, energy metabolism, dietary recommended intake.

## ESTIMATION OF PROTEIN REQUIREMENTS IN INDIAN PREGNANT WOMEN USING A WHOLE BODY POTASSIUM COUNTER (WBPC)

(1)Kuriyan Raj, Rebecca; (2)Naqvi, Saba; (2)Josephine D'Souza, Ninoshka; (3)Singh, Meenu; (4)Bhat, Kishor; (5)Mendiratta, Gaurav; (6)HPS, Sachdev; (7)Preston, Thomas; (8)Kurpad, Anura.

(1)Associate Professor. India.; (2)PhD student, Division of Nutrition. St. John's Research Institute (SJRI). Bangalore. India; (3)Post-doc student, Division of Nutrition. St. John's Research Institute (SJRI). Bangalore. India; (4)Assistant Professor, St. John's Research Institute (SJRI). Bangalore. India; (5)PhD student, Centre for High Energy Physics. Indian Institute of Science. Bangalore. India; (6)Professor, Sitaram Bhartia Institute of Science and Research. New Delhi. India; (7)Professor, Stable Isotope Biochemistry Laboratory. Scottish Universities Environmental Research Centre. Glasgow. Scotland. UK; (8)Professor, St. John's Research Institute (SJRI). Bangalore. India.

**Background and objectives:** The diets of Indian pregnant women do not often meet their increased gestational protein requirement, as defined by the WHO/FAO/UNU. Since the occurrence of low birth weight is high in India, it is critical to examine how the quantity/ quality of protein intake affects gestational body and fetal protein accretion. While body composition measurements during pregnancy are limited by hydration status and radiation exposure, measuring the naturally occurring radioactive isotope of potassium (40K) in the body, by a Whole Body Potassium Counter (WBPC), is independent of assumptions of the hydration status, and gives accurate estimates of Body Cell Mass (BCM) and further, the fat free mass (FFM). Accurate measurements of gestational BCM, as an indicator of protein accretion, will also define gestational protein requirements. The objective of the present study was to build a WBPC and use it to arrive at protein requirements for Indian pregnant women.

**Methods:** A state of the art WBPC was constructed in Bangalore, using a shadow shield, with alternate layers of steel and lead to shield four Sodium Iodide (NaI) crystal detectors from natural background radiation. Gamma spectra obtained were used to estimate total body potassium using phantoms of varying shapes/ sizes with known concentrations of potassium chloride and Monte Carlo simulations to adjust for body geometry. Fifty pregnant women were recruited in their first trimester and measurements of anthropometry, body composition and dietary intakes were performed at each trimester.

**Results:** The WBPC had an accuracy of 3%. Estimates of FFM from the WBPC correlated well with the FFM obtained by deuterium dilution and Dual Energy X-ray Absorptiometry (DEXA). The preliminary results suggest that the protein requirement is 14.2 ± 5.5 g/day in the 2nd trimester and 15.9 ± 7.0 g/day in the 3rd trimester, for a gestational weight gain of 12.5 kg in Indian women.

**Conclusions:** The results from this first facility in Asia will provide critical data with regard to protein requirement in pregnancy and will address several questions around pregnancy nutrition and birth outcomes in India.
**Keywords:** Body composition, whole body potassium counter, protein requirement, Indian pregnant women.

**Further collaborators:** Rebecca Kuriyan PhD, Saba Naqvi MSc, Ninoshka Josephine D'Souza MSc, Meenu Singh PhD, Kishor Bhat PhD, Gaurav Mendiratta PhD, HPS Sachdev MD, Thomas Preston PhD

Anura V Kurpad MD PhD.

144/2094

MATERNAL IRON ABSORPTION AND IRON TRANSFER TO THE FETUS DURING PREGNANCY IN NORMAL-WEIGHT AND OVERWEIGHT/OBESE WOMEN AND THE EFFECTS ON INFANT IRON STATUS

(1)Stoffel, Nicole Ursula; (2)Zimmermann, Michael Bruce; (3)Zeder, Christophe; (4)Quack-Loetscher, Katharina; (5)Herter-Aeberli, Isabelle.

(1)Msc. Department of Health Science and Technology. Laboratory of Human Nutrition. ETH Zürich. Switzerland.; (2)MD. Department of Health Science and Technology. Laboratory of Human Nutrition. ETH Zürich. Switzerland; (3)Msc. Department of Health Science and Technology. Laboratory of Human Nutrition. ETH Zürich. Switzerland; (4)MD. Department of Obstetrics. University Hospital Zürich. Switzerland; (5)PhD. Department of Health Science and Technology. Laboratory of Human Nutrition. ETH Zürich. Switzerland.

**Background and objectives:** Overweight/obesity causes low-grad systemic inflammation and thereby an up-regulation of the iron regulator hepcidin and a reduction in fractional iron absorption (FIA) even when iron stores are low. Pregnancy increases iron needs because of the expansion of maternal blood volume and fetal needs. It is unclear whether and/or to what extent overweight/obese pregnancy influences FIA, iron supply of the fetus and risk of iron deficiency in mother and newborn. In this case-control study, we (1) determined the impact of maternal overweight/obesity on FIA in pregnancy and on the transfer of iron to the fetus and newborn iron status; (2) confirmed the relationship between BMI, hepcidin, plasma ferritin (PF) and inflammatory markers; and (3) determined differences in women's iron needs during the 2nd and 3rd trimester.

**Methods:** In this ongoing multicenter case-control study (normal-weight n=36; overweight/obese n=36) we have administered labeled [57Fe]- or [58Fe]-FeSO4 to women during the 2nd and 3rd trimester of pregnancy. We measured FIA determining erythrocyte incorporation of iron stable isotopes 14 days after administration. From pregnancy week (PW) 12 until PW 36 iron-, inflammation and hepcidin were monitored. Iron transfer to the fetus was determined as iron stable isotope concentration in cord blood.

**Results:** Subject characteristics (mean±SD) in PW 12 for the normal-weight/obese were: age: 32±6/33±5years, BMI: 20.8±2.5/41.0±7.4kg/m2, hemoglobin: 12.1±0.9/13.5±0.8g/dL and PF: 61±23/47±24µg/L. Preliminary data showed an 83% and 24% lower FIA in the 2nd trimester compared to the 3rd in normal-weight and overweight/obese pregnant women, respectively.

Iron isotopes were highly detectable in cord blood. The [58Fe]/[57Fe]-ratio determined in cord blood corresponded to the [58Fe]/[57Fe]-ratio determined in the mother in the 3rd trimester.

**Conclusions:** In normal pregnancy, FIA increases over time to support increased iron needs of mother and fetus. This is consistent with decreasing hepcidin concentrations during pregnancy. Our preliminary data show a dramatically reduced increase in FIA in overweight/obese pregnant women indicating hepcidin may still play an important role in this group even in the 3rd trimester. Thus, even though iron demands are strongly increased, overweight/obesity may prohibit an adequate iron supply to the expecting mother and the fetus due to persistent subclinical inflammation.

**Keywords:** Overweight/obese pregnancy, hepcidin, subclinical inflammation, iron absorption, iron transfer to the fetus.

144/2501

RETENTION, IRON BIOAVAILABILITY AND SENSORY EVALUATION OF EXTRUDED RICE FORTIFIED WITH IRON, FOLIC ACID AND VITAMIN B12

(1)Madhari, Radhika Madhari; (2)WJ, Yvette; (3)Pullakhandam, Raghu; (4)P, Elumalai; (5)B, Swetha; (6)B, Naveen kumar; (7)T, Longvah.

(1)PhD. Nutrition Scientist. Division of Community Studies. National Institute of Nutrition. India; (2)MSc. Nutrition. Micronutrient Research Group. National Institute of Nutrition. India; (3)PhD. Micronutrient Research Group. National Institute of Nutrition. India; (4)Suvarnabhoomi Enterprises Private Limited. Tamil Nadu. India; (5)MSc. Nutrition. Division of Community Studies. National Institute of Nutrition. India; (6)PhD. Division of Biostatistics. National Institute of Nutrition. India; (7)MSc. Director Incharge. National Institute of Nutrition. India.

**Background and objectives:** Food fortification is considered a sustainable long term strategy for the prevention of micronutrient deficiencies in general population. Extrusion technologies enabled rice fortification with iron and other multiple nutrients. In the current study we have developed iron, folic acid and vitamin B12 fortified rice, and tested the retention, iron bioavailability and sensory properties compared to unfortified rice.

**Methods:** The rice flour is fortified with iron, folic acid and vitamin B12, and extruded rice kernels were prepared using a warm extrusion process. The iron, folic acid and vitamin B12 content were analyzed by HPLC methods before and after washing the fortified rice in water to test the retention. The iron dialyzability after simulated digestion of fortified rice was performed in a 6-well plate format. The iron content in rice, digesta and dialyzable fraction were estimated by atomic absorption spectrometry. The sensory evaluation was done in 84 adult human respondents using triangle test to evaluate if fortified rice was similar to unfortified rice both in the uncooked and cooked form.

**Results:** The iron (59.3mg±3.2/kg), folic acid (0.136±0.019mg/kg) and vitamin B12 (0.086±0.03 mcg/kg) of fortified rice, pre-
pared by 1:100 dilution of extruded rice mix with normal rice, is in agreement with expected fortification levels. The retention of iron, folic acid and B12 after incubation of fortified rice with excess water for a period of 1h, is more than 90% for all nutrients. The percent dialyzable iron from fortified rice (1.39%±0.3) was much lower compared to unfortified rice (6.79%±0.54), but net dialyzable iron content was 14 times higher in fortified rice. Triangle test revealed that 95% and 78% of the respondents were not able to distinguish the fortified rice from unfortified rice in uncooked (P=0.85) and cooked form (P=0.15) respectively.

**Conclusions:** These results suggest that rice fortified with iron, folic acid and vitamin B12 has high retention of nutrients during washing procedures at household levels, and provides more bioavailable iron compared to unfortified rice. Further, sensory studies also demonstrated acceptable similarity between fortified and unfortified rice and therefore can be used interchangeably. Nevertheless, further studies in human subjects are required to validate these in vitro findings.

**Keywords:** Food fortification, extruded rice, iron, folic acid, vitamin B12, bioavailability, sensory properties.

**Conflict of Interest Disclosure:** The project is funded by Suvarnabhoomi Enterprises Pvt. Ltd., Tamil Nadu, India and the authors declare no conflict of interest.

144/2768

IS ENERGY EXPENDITURE CONSIDERED IN THE LITERATURE WHEN ENERGY INTAKE IS MEASURED? A NEED FOR A METHODOLOGICAL CONSENSUS

(1)González-Gross, Marcela; (2)Aparicio Ugarriza, Raquel; (2)Calonge-Pascual, Sergio; (3)Gómez-Martínez, Sonia; (2)García-Carro, Alberto;

(4)Sanz-Valero, Javier; (5)Martínez, J. Alfredo; (6)Gil, Ángel; (7)Marcos, Ascensión; (8)Moreno, Luis A.

(1)PhD. Professor. Universidad Politécnica de Madrid. Vice-president. Spanish Nutriiton Society (SEÑ). Madrid. Spain.; (2)MSc. ImFINE Research Group. Department of Health and Human Performance. Universidad Politécnica de Madrid. Spain; (3)PhD. Instituto de Ciencia y Tecnología de Alimentos y Nutrición (ICTAN). Consejo Superior de Investigaciones Científicas. Madrid. Spain; (4)PhD. Professor. Departamento de Salud Pública. Universidad Miguel Hernández. Alicante. Spain.; (5)PhD. Professor. Dept. of Nutrition, Food Science, Physiology Navarra University. Spain; (6)PhD. Professor. Departamento de Bioquímica y Biología Molecular II. Universidad de Granada. Spain.; (7)PhD. Professor. Instituto de Ciencia y Tecnología de Alimentos y Nutrición (ICTAN). Consejo Superior de Investigaciones Científicas. Madrid. Spain; (8)PhD. Professor. Universidad de Zaragoza. President of the Spanish Nutrition Society (SEÑ). Spain.

**Background and objectives:** One of the health-transitions humans have passed during the XX Century related to nutrition is without doubt that from undernutrition to obesity, which persists in the ongoing years of the XXI Century. Energy intake (EI) is a contributing factor and therefore a "hot topic" in nutritional

sciences. The aim of this study was to review if studies including data on dietary intake, and specifically EI, also included data on energy expenditure (EE) and physical activity (PA).

**Methods:** A systematic search of PubMed from 1953 to December 2015 was conducted. Our systematic search consisted of keywords related to energy intake and nutrition ("nutritional requirements", "recommended dietary allowances" or "diet records"). We used MeSH and MajR terms to build up a structured search and the final equation was: ("Recommended Dietary Allowances"[Majr] OR "Nutritional Requirements"[Majr] OR "Energy Intake"[Majr] OR "Diet Records"[Majr]) NOT ("Diseases Category"[Mesh] OR "Pregnancy"[Mesh] OR "Breast Feeding"[Mesh]). Filters were "Humans" and "Adults over >19 years".

**Results:** From 2229 identified articles, 933 articles were finally retained fulfilling inclusion and quality criteria. A total of 307543 adults were included, and most studies had been performed in EEUU, UK, Canada, Australia, Japan and Europe (mainly France, followed by Ireland and Denmark); only 9 studies were conducted in Spain. Few studies took place in Africa in comparison with other continents. Only 15% of the studies included both EI and EE. A great methodological variety was observed, with more than 14 different methods regarding EI, and more than 10 for EE. PA was only analysed in few studies, and mostly not considered for interpretation of data and conclusions. Moreover, PA was commonly measured by non- validated questionnaires.

**Conclusions:** A great diversity regarding the methodology was observed, which makes comparability among studies difficult. EE and PA are missing in around 80% of studies or not included in interpretation of results. Conclusions regarding EI in adults should not be taken without interpreting EE. We highlight the need for a methodological consensus and analyses standardization regarding dietary intake and energy expenditure in nutritional studies.

**Keywords:** Energy intake, energy expenditure, physical activity, energy balance.

**Further collaborators:** Spanish Nutrition Society

144/2887

THE PREVALENCE OF MALNUTRITION IN ENRICH PROJECT SITES IN BANGLADESH, KENYA, MYANMAR, PAKISTAN AND TANZANIA

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

(1)MD.MPH. Chief of Party. ENRICH Program. World Vision Canada. Canada; (2)Grant/DME advisor. World Vision Canada. Canada;

(3)DME Team Leader. World Vision Canada. Canada.

**Background and objectives:** ENRICH is a Maternal New-born and Child Health and Nutrition program funded by the Government of Canada and implemented in Bangladesh, Pakistan, Myanmar, Kenya and Tanzania. ENRICH will use nutrition pathway to reduce U5M by strengthening health systems and improving basic nutrition and nutrition sensitive services and addressing 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 assessment was done as part of the project baseline survey. Stratified multi-stage cluster sampling method was used to select households and respondents. ENA for SMART software was used to determine sample size and for data analysis. Children between 6-59 months were measured to determine their nutrition status. Mothers/caregiver's knowledge on Infant and Yong Child Feeding practices and status of household food security were also assessed using a structured questionnaire.

**Results:** The prevalence of stunting in ENRICH project sites in Bangladesh, Kenyan, Myanmar, Pakistan and Tanzania was 30.9%, 35.1% 39.0%, 33.8% 31.5% respectively. Prevalence of wasting was 6.5%, 5.3%, 7.0%, 10.2% and 4.9% in the same order. Undernutri- tion was also ranged from as low as 15.3% in Tanzania to 29.7% in Pakistan. The proportion of households with sever food insecurity in Bangladesh, Kenyan, Pakistan and Tanzania was 11.0% 23.0%, 15.5%, 32. 3% respectively. Food insecurity and poor infant and young child feeding practices are among the major underlying causes of malnutrition in the target communities.

**Conclusions:** Malnutrition is a major public health problem in implementing countries. Integrated nutrition specific and sen- sitive interventions are required to address the immediate and un- derlying causes of child malnutrition in the target communities. The project will implement food based interventions such as bio- fortification to improve availability of nutrient dense foods

**Keywords:** Stunting, Wasting, biofortification.

**Further collaborators.**

World Vision Implements ENRICH project in collaboration with Nutrition International and HarvestPlus.

Twice repeated samplings of 24-h urine and spot urine samples were collected to estimate habitual daily iodine intake of children. Thyroid volume of children was measured and blood samples were collected to determine thyroid function indicators.

**Results:** This study enrolled 2224 children including 1100 boys and 1114 girls. The median iodine content of water in this study was 181 (67.2-402) µg/L. In the first sampling, the median urinary iodine concentration (UIC) of spot urine and 24-h urine (24-h UIC) were 481 (218-818) µg/L and 380 (203-646) µg/L, while the spot UIC and 24-h UIC were 417 (195-753) µg/L and 397 (200-682) µg/L, respec- tively, indicating iodine status of children in this study was excessive. The habitual iodine intake (Best linear unbiased predicator, BLUP) of children was 298 (186-437) µg/d. The total goiter rate (TGR) was 9.7% and the incidence of subclinical hypothyroidism (SCH) was 237 (11.5%) in children. TSH of children increased as the increase of iodine intake (r=0.11, P<0.0001), while the incidence of SCH was 11.2% in children with iodine intake of 200-300µg/d and increased accordingly. Adjusted for age, sex, height, weight, logistical regres- sion analysis shows that the risk of TGR significantly increased at iodine intake of 250-299µg/d in 7-10 year-old children and the risk of TGR increased at iodine intake of 300-399µg/d in 11-14 year- old children, and the risk increased as the increase of iodine intake. However, there were no differences in the risk of SCH among differ- ent iodine intake groups (P>0.05).

**Conclusions:** TGR may be a more sensitive indicator to reflect long-term iodine excess. We recommend the UL of iodine for 7-10 year-old children is 250µg/d and the UL of iodine for 11-14-year- old children was 300µg/d.

**Keywords:** Children, tolerable upper safe intake level, thyroid volume, subclinical hypothyroidism (SCH).

**Further collaborators.**

Elizabeth N Pearce, MD., Section of Endocrinology, Diabetes, and Nutrition, Boston University School of Medicine, Boston, MA, USA

Michael B Zimmermann, PhD. , the Human Nutrition Labo- ratory, Swiss Federal Institute of Technology, Zu¨rich, Switzerland (Michael B Zimmermann, PhD)

144/2898

ADVERSE EFFECTS ON THYROID OF CHINESE CHILDREN EXPOSED TO LONG-TERM IODINE EX- CESS: OPTIMAL AND SAFE UPPER INTAKE LEVEL OF IODINE FOR 7-14-YEAR-OLD CHILDREN

(1)Zhang, Wanqi; (2)Chen, Wen; (2)Wang, Wei; (3)Du, Cong; (3)Wu, Yalan;

(4)Bian, Jiancao; (3)Shen, Jun; (2)Lin, Laixiang; (2)Tan, Long; (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:** The adverse effects of iodine excess on thyroid in children were not well understood. This study aimed to assess the effects of iodine intake on thyroid and explore the safe iodine intake level (UL) for children.

**Methods:** A multi-stage cross-sectional study involved 2224 children from areas with high iodine content in drinking water.
# Track 6: Functional Foods and Bioactive Compounds

144/267

## BIOINFORMATIC AND CHEINFORMATIC APPROACHES IN STUDYING BIOACTIVE PEPTIDES DERIVED FROM FOOD PROTEINS

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

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

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

**Background and objectives:** There are few criteria to evaluate proteins originating from foods. One of them is result-ed from the presence of peptide fragments generated upon the proteolytic enzymes action. These fragments are called bioactive peptides (biopeptides) because they can regulate body functions. Many peptides act as inhibitors of angiotensin converting en-zyme (ACE inhibitors), antioxidant, antibacterial, antithrombot-ic etc. agents. as well as affect the quality of foods due to their taste properties.

The progress in the development of information technologies contributed to the massive growth of scientific interests in the ap-plication of computer methodologies in studying bioactive pep-tide sequences. These methodologies concerning the analysis of biopeptides include e.g.: elaboration of databases of information, development of criteria of evaluation of proteins as the potential source of bioactive peptides, and/or programs to simulate protein hydrolysis to generate biopeptides. This bioinformatic methods are suitable to study peptides especially when combined with the cheminformatic approach including the databases of chemical information about peptides as well as multivariate analyses like principal component analysis (PCA), partial least square meth-ods (PLS) and artificial neural networks (ANN). Such approach is useful in prediction of relationship between the structure and bioactivity of peptides (QSAR approach).

**Methods:** The aim of the study was to present the method-ology involving some examples of bioinformatic/cheminformat-ic analysis of peptides derived from food proteins. This approach includes the presentation of sources of information on peptides provided in bioinformatic/cheminformatic databases like BIO-PEP, EropMoscow, ChemSpider or ChEMBL. Moreover, some examples of cheminformatics methods to study the properties of peptides in the context of their bioactivity will be presented.

**Results:** The results of such studies can be useful when com-bined with the research concerning generation of peptides with use of commercial enzymes or human digestive juices.

**Conclusions:** The approach presented above enables develop-ing the knowledge on peptides derived from different foods.

**Keywords:** Bioactive peptides, proteins, bioinformatics, cheminformatics.

144/402

## GINSENG OLIGOPEPTIDES PROTECT RATS AGAINST BINGE DRINKING-INDUCED LIVER INJURY

(1)Li, Yong; (2)Liu, Rui; (3)Mao, Ruixue; (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:** The present study aimed to investigate the effects and mechanism of ginseng oligopeptide (GOP) on acute alcohol poisoning in rats.

**Methods:** A total of 70 SD rats were randomly assigned to 7 groups, including a normal control group, a model group, a whey protein group (0.2500 g/kg), and 4 GOP intervention groups (0.0625, 0.1250, 0.2500, 0.5000 g/kg). 7 g/kg/BW 50% v ethanol was given intragastrically after the 30 days of rat stomach-gavaged continuously. The serum aminotransferase and inflammatory cy-tokine were assayed, the alcohol dehydrogenase (ADH) activity and oxidative stress and lipid peroxidation parameters in liver tis-sue were conducted.

**Results:** We found that the GOP treatment reduced the levels of TNF-α, IL-6, IL-1β in serum and MDA concentrations in liver (p<0.05), increased the levels of AST, ALT in serum and the con-tents of ADH, SOD, GSH and GSH-PX in liver tissue(p<0.05).

**Conclusions:** These findings suggest that GOP have a signifi-cant protective effect on acute alcohol poisoning in rats and acute liver injury, and the underlying mechanism may be related to in-hibit the inflammatory response and Lipid peroxidation reaction, and to improve the alcohol dehydrogenase activity and enhance anti-oxidative activities.

**Keywords:** Ginseng oligopeptide; Binge Drinking-Induced Liver Injury; Oxidative stress; Lipid peroxidation; Inflammatory response.

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

144/416

## EICOSAPENTAENOIC ACID AND DOCOSAHEXAENOIC ACID HAVE ANTIDEPRESSANT EFFECTS WITH 17B-ESTRADIOL INJECTION VIA REGULATION OF A NEUROBIOLOGICAL SYSTEM IN OVA-RIECTOMIZED RATS

(1)Park, Yongsoon; (2)Choi, Jeongeun.

(1)PhD. Professor. Department of Food and Nutrition, Hanyang University. Seoul. Korea; (2)MSc. Graduate Student. Department of Food and Nutrition. Hanyang University. Seoul. Korea.

**Background and objectives:** Our previous studies found that n-3 polyunsaturated fatty acids (PUFAs) and estrogen had
synergistic antidepressant-like effects. The purpose of the pres-
ent study was to investigate the hypothesis that three major n-3
PUFAs, α-linolenic acid (ALA), eicosapentaenoic acid (EPA) and
docosahexaenoic acid (DHA), individually had antidepressant ef-
fects combined with 17β-estradiol-3-benzoate (E) through a neu-
robiological pathway in ovariectomized (OVX) rats.

**Methods:** The experimental protocol was approved by the Institutional Animal Care and Use Committee of Hanyang University (HY-IACUC-16-0044). Rats were fed a modified AIN-93G diet with 0% n-3 PUFAs, and 1% ALA, EPA and DHA relative to total energy intake for 12 weeks, and were injected with corn oil or E every 4 days during the last 3 weeks.

**Results:** Regardless of E injection and sham surgery, supplementation with ALA increased the brain phospholipid proportion of 18:3n3 as compared to the NC diet and supplementation with EPA and DHA. Supplementation with ALA, EPA and DHA increased the brain phospholipid proportions of 20:5n3, 22:5n3 and 22:6n3; this increase was greater with EPA and DHA than ALA supplementation. Supplementation of EPA, DHA and E increased serum concentrations of serotonin and climbing behavior, and decreased immobility during a forced swimming test. Supplementation with EPA, DHA and E also decreased hippocampal expressions of interleukin-6 and tumor necrosis factor-α, and increased cAMP response element binding protein, brain-derived neurotrophic factor (BDNF) and estrogen receptor-α. Immunofluorescence staining consistently showed elevated expressions of BDNF. Magnetic resonance spectroscopy showed that E increased glucose and decreased glutamate, glutamine and myo-inositol concentrations regardless of n-3 PUFA supplementation. In addition, supplementation with EPA, DHA and E decreased levels of nitric oxide metabolite. However, ALA had no antidepressant effect.

**Conclusions:** The present study suggested that the antidepressant-like effects of EPA and DHA supplementation and E injection could be due to the regulation of serotonergic neurotransmission and inflammatory cytokines, rather than due to the anti-oxidative system. Supplementation with n-3 PUFA and E had the additional function of modulating neurometabolites in the hippocampus.

**Keywords:** BDNF. Depression. Estrogen. Neurometabolite. N-3 polyunsaturated fatty acids.

**Further collaborators:** This study was supported by a Korean Research Foundation grant funded by the Korean Government (NRF-2015R1D1A1A09060823).

**Background and objectives:** Alzheimer's disease (AD) is the most common age-dependent neurodegenerative disorder and causes a progressive decline in cognitive function. Therefore, it has been highly desirable to find a method of preventing and treating AD. 1'-Acetoxychavicol acetate (ACA) is naturally obtained from the rhizomes and seeds of Alpinia galangal. The plants are used as a spice or Chinese medicine in Southeast Asia. ACA exhibits various biological activities. However, the effect of ACA on neuronal cells death remains unclear. Here, we examined the effect of ACA on learning and memory in senescence-accelerated mice prone 8 (SAMP8).

**Methods:** Eight age-matched SAM resistance/1 (SAMR1) mice and 24 SAMP8 mice (8 weeks old) were used. After the interval (2 weeks) of acclimation, SAMR1 mice were used as homologous controls and 24 SAMP8 mice were assigned randomly to the control group and the 0.02% ACA group and maintained for 25 weeks. After 25 weeks, the learning ability in the Morris water maze test and the Y-maze test were performed. Serum metabolite profiles were obtained by GC-MS analysis, and each metabolic profile was plotted on a 3D score plot.

**Results:** In mice that were fed a control diet containing 0.02% ACA, the learning ability in the Morris water maze test was significantly enhanced in comparison with mice that were fed the control diet alone. In the Y-maze test, SAMP8 mice showed decreased spontaneous alterations in comparison with senescence-accelerated resistant/1 (SAMR1) mice, a homologous control, which was improved by ACA pretreatment. Furthermore, the contents of betab-hydroxybutyric acid and palmitic acid in the serum of SAMP8-ACA mice were higher than those of SAMP8-control mice and SAMR1-control mice. We also found that SAMR1 mice did not show histological abnormalities, whereas histological damage in the CA1 region of the hippocampus in SAMP8-control mice was observed. However, SAMP8-ACA mice were observed in a similar manner as SAMR1 mice.

**Conclusions:** These findings confirm that ACA increases the serum concentrations of betab-hydroxybutyric acid and palmitic acid levels and thus these fuels might contribute to the maintenance of the cognitive performance of SAMP8 mice.

<footnote>(1)Kojima-Yuasa, Akiko; (2)Yamamoto, Tomiya; (3)Yaku, Keisuke;</footnote>

<footnote>(2)Hirota, Shiori; (4)Matsui-Yuasa, Isao.</footnote>

<footnote>(1)PhD. Associate Professor. Department of Food and Human Health Sciences. Graduate School of Human Life Science. Osaka City University. Osaka. Japan.; (2)MSc. Graduate student. Department of Food and Human Health Sciences. Graduate School of Human Life Science. Osaka City University. Osaka. Japan.; (3)PhD. Department of Food and Human Health Sciences. Graduate School of Human Life Science. Osaka City University. Osaka. Japan.; (4)DSc & PhD. Professor. Department of Food and Human Health Sciences. Graduate School of Human Life Science. Osaka City University. Osaka. Japan.</footnote>
**Keywords:** Morris water maze test; Y-maze test; 1'-Acetoxychavicol acetate; Senescence-accelerated mouse prone 8 (SAMP8); Betab-hydroxybutyric acid.

144/675

BREAD FORTIFICATION WITH MICROENCAPSULATED VITAMIN D

(1)Leiva Rodriguez, Carla Andrea; (2)Benavides, Sergio; (3)Franco, Wendy; (1)Rojas, Loreto.

(1)Msc. Nutricionista. Carrera de Nutrición y Dietética. Pontificia Universidad Católica de Chile. Chile; (2)PhD (C). Ingeniero en Alimentos. Facultad de Ingeniería Química y Bioprocesos. Pontificia Universidad Católica de Chile. Chile; (3)PhD. Ingeniero en Alimentos. Facultad de Ingeniería Química y Bioprocesos. Pontificia Universidad Católica de Chile. Chile.

**Background and objectives:** Vitamin D plays an important role in the health of the population, regarding bone metabolism, cardiovascular system, insulin resistance, diabetes, neurological development, immunomodulation and regulation of cell growth. Active Vitamin D can be derived from the skin, where 7-dehydrocholesterol or diet is formed from its precursor by the action of solar radiation. The evidence reveals that at national and international level, there is a generalized deficiency of Vitamin D. An alternative to diminish this deficiency is to incorporate vitamin D in food of habitual consumption. In our country, fortified wheat flour proves to be a good vehicle since it is of regular consumption and is considered a food of first necessity by the school and adult population. However, the incorporation of vitamin D into food matrices presents a disadvantage, since the thermal treatments that undergo these matrices can limit its bioavailability and bioactivity. Because of the above, microencapsulation is a very good alternative to protect vitamin D for later use in food processing.

The objectives of this research were to evaluate the effect of concentration and degree of dispersion of vitamin D on the quality of the process of encapsulation by ionic gelation.

**Methods:** Emulsions were prepared from alginate solutions incorporating vitamin D in three concentrations (1, 2 and 3% v / v). Emulsions were prepared at three speeds and a dispersion time of 5 min. Emulsions were evaluated by image analysis. The drop size of the dispersed phase was determined as well as the polydispersity. To obtain the microcapsules, each emulsion was dripped into a solution of CaCl2 (1.0% w / v). The microcapsules were oven dried at 35 °C for 12 hours. The efficiency of encapsulation and load capacity by spectrophotometry was determined.

**Results:** The results indicated that EE% and CC% are dependent on the concentration and degree of dispersion of vitamin D. In addition, it was determined that vitamin D concentration affects the size and shape of the microcapsule.

**Conclusions:** Therefore microencapsulation is a viable alternative for vitamin D. Future studies will analyze the organoleptic quality of bread and the bioavailability of microencapsulated Vitamin D.

**Keywords:** Vitamin D, fortification.

144/686

STIGMASTEROL SERUM ELECTROLYTES ELEVATING POTENTIALS IN POLOXAMER-407 INDUCED HYPERLIPIDEMIC ALBINO RATS

(1)Iombor, Terhemba; (2)Nnam, Ngozi.

(1)MSc. Nutritionist. Department of Home Science and Management. University of Agriculture. Makurdi. Nigeria.;

(2)PhD. Nutritionist. Department of Home Science, Nutritio and Dietetics. University of Nigeria.Nsukka. Nigeria.

**Background and objectives:** Hyperlipidemia often occurs in conjunction with other metabolic risk factors including glucose intolerance, obesity, diabetes and metabolic syndromes. The study investigated effect of stigmasterol administration on serum calcium, potassium, sodium and phosphorus levels of poloxamer-407 induced hyperlipidemia in albino rats.

**Methods:** Thirty albino rats were divided into six groups of five rats viz: normal, poloxamer, atorvastatin and 50, 100 and 200 mg stigmasterol treated animals. Poloxamer-407 was dissolved in saline (1g/mL, 40C) and administered by intraperitoneal injection at a dose of 1g/kg body weight. Injections were given at days 1 and 3. Blood samples were obtained by cardiac puncture at baseline, day 4 and post-treatment.

**Results:** Induction of hyperlipidemia significant decreased serum calcium in positive control (4.52-4.41 mg/dl), atorvastatin (4.34-4.18 mg/dl), 50 mg (5.08-4.46 mg/dl), 100 mg (4.46-4.10 mg/dl) and 200 mg (4.83-4.35 mg/dl) stigmasterol treated rats. Induction serum potassium decreased from 0.69-0.65 mg/dl (positive control), 0.85-0.82 mg/dl (atorvastatin), 0.90-0.79 mg/dl (50 mg), 0.82-0.68 mg/dl (100 mg) and 0.87-0.72 mg/dl (200 mg) stigmasterol treated rats. Induction serum sodium decreased from 55.23-54.97 mg/dl (positive control), 67.13-66.97 mg/dl (atorvastatin), 74.09-65.39 mg/dl (50 mg), 74.28-66.16 mg/dl (100 mg) and 66.35-65.16 mg/dl (200 mg) stigmasterol treated rats. Induction serum phosphorus decreased from 3.38-3.27 mg/dl (positive control), 4.60-4.24 mg/dl (atorvastation), 4.44-4.20 mg/dl (50 mg), 3.81-3.36 mg/dl (100 mg) and 3.51-3.10 mg/dl (200 mg) stigmasterol treated rats. Post-treatment serum calcium increased from 4.18-5.39 mg/dl (atoevastatin), 4.46-5.68 mg/dl (50 mg), 4.10-5.24 mg/dl (100 mg) and 4.35-5.72 mg/dl (200 mg) stigmasterol treated rats. The serum potassium levels at post-treatment increased from 0.82-0.85 mg/dl (atorvastatin), 0.79-0.90 mg/dl (50 mg), 0.68-0.82 mg/dl (100 mg) and 0.72-0.87 mg/dl (200 mg) stigmasterol treated rats. Post-treatment serum sodium increased from 66.97-122.91 mg/dl (atorvastatin), 65.39-123.65 mg/dl (50 mg), 66.16-127.71 mg/dl (100 mg) and 65.16-122.42 mg/dl (200 mg) stigmasterol treated rats. Post-treatment serum phosphorus increased from 4.18-5.51 mg/dl (atorvastatin), 3.69-4.57 mg/dl (50 mg), 4.28-5 mg/dl (100 mg) and 3.52-5.18 mg/dl (200 mg) stigmasterol treated rats.

**Conclusions:** Stigmasterol has electrolytes elevating potential in a dose dependent manner.

**Keywords:** Electrolytes, Stigmasterol, Poloxamer-407, Hyperlipidemia, Atorvastatin.
# GLYCOGEN IMPROVES COGNITIVE FUNCTION IN HUMAN AND MICE

Furuyashiki, Takashi; Kato, Kazuko; Kakutani, Ryo; Kamasaka, Hiroshi; Kuriki, Takashi.

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

**Background and objectives:** Recently, we have developed the method of synthesizing glycogen from corn starch by using 3 enzymes, isoamylase, branching enzyme and amylomaltase. We named the product enzymatically synthesized glycogen (ESG), which is equivalent in the physicochemical properties to natural-source glycogen. It is confirmed that ESG possess health-beneficial functions such as immunomodulating effect and dietary fiber like effect. In this study, the effects of ESG on brain function were investigated in human clinical study and animal study.

**Methods:** Human clinical study: The study was approved by the ethics committee of Ezaki Glico Co., Ltd. and carried out with a placebo-controlled, double-blind, randomized, cross-over design. Thirty-eight healthy adult subjects ingested 5 g/day of ESG or maltodextrin as a placebo for 4 weeks. Before and after 2 and 4 weeks of starting ingestion, cognitive performances were detected by using CogHealth battery. Subjective fatigue and stress markers in saliva were measured simultaneously.

Animal study: The study was approved by the Institutional Animal Care and Use Committee of Ezaki Glico Co., Ltd. and performed in accordance with the Guidelines for Proper Conduct of Animal Experiments (Science Council of Japan). Senescence-accelerated mouse prone 8 (SAMP8) were fed 6.0% ESG containing feed or AIN-93M as a control for 6 month. After that, memory function was evaluated by Morris water maze.

**Results:** In the human clinical study, supplementation of ESG significantly improved the visual discrimination at 2 week and the memory function at 4 week compared with placebo control. On the other hand, subjective fatigue and stress markers were not changed by ESG. In the study of SAMP8 mice, the better memory function was observed in ESG ingestion group.

**Conclusions:** In this study, we confirmed that ESG ingestion improves cognitive function in human and mice.

**Keywords:** Glycogen, brain, cognitive function, human, mice

# SUPPRESSION OF POSTPRANDIAL HYPERGLYCEMIA BY BUCKWHEAT ALBUMIN

(1)Kumagai, Hitomi; (2)Yamaguchi, Yusuke; (2)Ninomiya, Kazumi;

(2)Ina, Shigenobu; (2)Akao, Makoto; (3)Kumagai, Hitoshi.

(1)Nihon University; (2)Department of Chemistry and Life Science, Nihon University, Fujisawa-shi, Japan; (3)Professor, Department of Food Science and Nutrition, Kyoritsu Women's University, Chiyoda-ku, Tokyo, Japan.

**Background and objectives:** Diabetes mellitus is a disorder of blood glucose regulation. Most of the patients are categorized into type 2 that shows a symptom of insulin resistance and/or deficiency. For the prevention of type 2 diabetes, it is important to control the postprandial blood glucose level by an appropriate food intake. The retardation of starch hydrolysis by inhibiting the α-amylase activity is one of the effective ways to suppress the elevation in blood glucose level. Wheat α-amylase inhibitor possesses the inhibitory activity against α-amylase and suppresses postprandial hyperglycemia after starch administration. It has been used as a functional component in Food for Specified Health Uses (FOSHU) in Japan. We have already reported that rice albumin is indigestible and effectively suppresses the elevation in blood glucose level not only after starch administration but also after glucose administration. However, the effect of other cereal protein on hyperglycemia has not been fully examined yet. Therefore, the present study was conducted to examine if buckwheat protein could suppress postprandial hyperglycemia.

**Methods:** Albumin was purified from buckwheat by ammonium-sulfate precipitation and gel filtration. First, the inhibitory activity of buckwheat albumin against α-amylase from human saliva, porcine pancreas and mealworm was evaluated in vitro. Next, rats were administered starch with and without the addition of buckwheat albumin. After the administration, the blood was collected from the tail vein at certain intervals. Then, the blood glucose level and the plasma insulin level were measured.

**Results:** Buckwheat albumin inhibited the activity of α-amylase from porcine pancreas and mealworm, but did not inhibit that of α-amylase from human saliva. Although buckwheat albumin was hydrolyzed by digestive enzymes, it suppressed the elevation in blood glucose level after carbohydrate administration. The plasma insulin level was also suppressed by the administration of buckwheat albumin.

**Conclusions:** Buckwheat albumin suppressed postprandial hyperglycemia in vivo though it was digestible indicating that peptides in the hydrolysate maintained the inhibitory activity against α-amylase even after digestion.

**Keywords:** Inhibition of α-amylase activity, suppression of postprandial hyperglycemia, buckwheat albumin, diabetes mellitus.
# ADDRESSING FOOD, NUTRITION AND ECO-
NOMIC SECURITY THROUGH IRON FORTIFIED
RICE TECHNOLOGY

(1)Angeles-Agdeppa, Imelda; (2)Saises, Marcela; (3)Capanzana, Mario.

(1)Assistant Scientist. Food and Nutrition research Institute, Department of Science and Technology, Taguig City, Philippines;

(2)Senior Research Specialist. Food and Nutrition Research Institute, Department of Science and Technology, Taguig City, Philippines; (3)Director. Food and Nutrition Research Institute, Department of Science and Technology, Taguig City, Philippines.

**Background and objectives:** The Philippine Food Fortification Law covers mandatory fortification of staples like rice with iron due to the alarmingly high prevalence of iron deficiency anemia (IDA) in 1998 (30.6%). Rice was used as a vehicle for fortification but was projected to be inadequate in 2025 to supply the growing population. During the milling process the recovery rate is estimated at 60% whole grain and 40% broken grain. Objective: This paper presents the results of the different studies of FNRI on iron-fortified rice addressing pronged issues: food, nutrition and economic insecurities besetting the Filipino population.

**Methods:** FNRI conducted Series of studies on rice fortification: (1) technology generation and product development of iron rice premix (IRP) and iron fortified rice (IFR); (2) efficacy trial; (3) pilot-scale field trials; (4) large-scale commercialization of IFR; and (5) scaling-up in Mindanao and national implementation.

**Results:** FNRI developed IRP from broken grains made into flour and blended with iron using hot extrusion technology which is stable for 24 months. The IRP is mixed with ordinary rice to produce IFR. An efficacy study among schoolchildren showed significant decline in anemia prevalence from 100% to 33%. The market trials studies in the municipality of Orion, Bataan and the province of Zambales showed that IFR is efficacious in reducing anemia and feasible to be sold in public markets. The FNRI has scaled-up the transfer of the IRP and IFR technologies to several private mills and investors. Currently, IFR is used in the Social Safety Net (SSN) programs of the government.

**Conclusions:** The technology of IRP addresses the 3 pronged issues: (1) food security by increasing rice supply because the broken rice which is usually used for animal feed is converted to whole grain rice for human consumption; and (2) nutrition security by increasing iron intake through fortification, and (3) economic security by converting broken rice which is a low priced rice (PhP 27.00) to high priced whole grain rice (Php 45.00). The proven significant benefits of fortifying rice merits to be scaled up to more millers and investors to make available ample supply of IFR for nationwide consumption.

**Keywords:** Food, nutrition, economic security, iron fortified rice.

**Further collaborators:** ILSI-CHP Japan.

# ANTIOXIDANT ACTIVITY AND AVAILABILITY OF POLYPHENOLS IN FRUITS AND VEGETABLES OF MAJOR CONSUMPTION IN THE PERUVIAN POPULATION

(1)Fernández Condori, Roxana; (2)Ramos-Escudero, Fernando.

(1)MSc. Nutritionist. Roxana Fernández Condori. Carrera Profesional de Nutrición y Dietética. Facultad de Ciencias de la Salud. Universidad Católica Sedes Sapientiae-UCSS. Perú; (2)PhD. Chemistry. Carrera de Nutrición y Dietética. Facultad de Ciencias de la Salud. Universidad Peruana de Ciencias Aplicadas-UPC. Perú.

**Background and objectives:** In fact, a big variety of fruits and vegetables which differ in shape, color, taste and nutritional value are available in the market of the Peruvian population. In this study, the physicochemical characteristics (pH, °Brix and acidity), total phenol content and antioxidant activity of fruits (strawberry, grape, papaya, banana "seda", mango, apple "Israel", watermelon, orange, tangerine, lemon) and vegetables (celery, cabbage, pumpkin, tomato, garlic, onion, Peruvian yellow chili, carrot, baby corn, lettuce) were evaluated.

**Methods:** The fruits and vegetables were analyzed using physicochemical analysis (volumetric titrations, potentiometric and refractometry methods). The total polyphenol content by the Folin-Ciocalteu method and antioxidant activity by QUENCHER-CUPRAC method (visible molecular absorption spectroscopy).

**Results:** The strawberry and grape showed the highest total phenolic content in fruits while onion and Peruvian yellow chili showed higher content of polyphenols in vegetables. The highest antioxidant activity in the QUENCHER-CUPRAC assay was found for strawberry, grape, apple "Israel", orange and tangerine while the showed highest antioxidant activity in vegetables was for garlic, onion and Peruvian yellow chili. Additionally, the positive correlation between the phenolic compounds content and antioxidant activity of the studied samples has been noted. However, the linear correlation between the polyphenol content versus the antioxidant activity in fruits was higher than in the vegetables.

**Conclusions:** Our results revealed that the fruits presented highest availability of polyphenols and a great antioxidant activity in compared to vegetables.

**Keywords:** Fruits, vegetables, polyphenols, antioxidant activity, peruvian populations.

**Conflict of Interest Disclosure:** The authors have no conflicts of interest to declare. This study is the doctoral thesis of RFC that has been developed within the Doctoral Program in Biochemistry and Nutrition at the university San Martin de Porres, Lima, Peru
## OXIDATION OF MAJOR CATECHINS IN GREEN, BLACK AND OOLONG TEAS AND THEIR CORRESPONDING BIOACTIVITIES

(1)Jiang, Heyuan; (2)Engelhardt, Ulrich H.; (3)Zhang, Jiangyong;

(3)Wang, Weiwei; (4)Shi, Liting; (4)Su, Wei.

(1)Prof/PhD.Scientist.Tea Research Institute of Chinese Academy of Agricultural Sciences.China; (2)Prof/PhD.Institute of Food Chemistry of Technische Universität Braunschweig.Germany;

(3)Assit Prof.Tea Research Institute of Chinese Academy of Agricultural Sciences.China; (4)Master Candidate.Tea Research Institute of Chinese Academy of Agricultural Sciences.China.

**Background and objectives:** Tea (Camellia sinensis L.O. Kuntze) is the most popular flavored and healthy beverage beyond water in the world. Catechins are the most important compounds in tea due to their physiological activity and the important effect on tea brew color and taste. In different kinds of teas, catechins might be kept, or be oxidized into dimers and other polymers, and could exhibit different bioactivities. This paper will figure out the major catechins oxides, and their corresponding bioactivities.

**Methods:** In this paper, we compared the content and composition of catechins and their major dimers including theaflavins and theasinensins, in green, black and oolong teas, based on UH-PLC and HPLC assisted with electrospray tandem MS. The bioactivities of catechin oxides and the related tea samples were also analyzed based on in-vitro studies.

**Results:** Besides green teas as non-fermentated teas, catechins encounters different degrees of catechins oxidation, whilst different kinds of oxidation pathways to form some kinds of catechin oxides, in black teas or oolong teas respectively as full- or partial- fermentated teas. **Results:** **Conclusions:** Their compositions might induce differential biological activities from those tea samples due to variant oxidation pathways of catechins.

**Keywords:** Tea, catechin dimers, theaflavins, theasinensins, bioactivities.

## ACCURATE QUANTITATION OF PLASMALOGEN AND RELATED PHOSPHOLIPIDS BY USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY COUPLED WITH MASS SPECTROMETRY

(1)Otoki, Yurika; (2)Kato, Shunji; (3)Miyazawa, Teruo; (2)Nakagawa, Kiyotaka.

(1)PhD. Agriculture. Graduate School of Agricultural Science. Tohoku University. Japan.; (2)PhD. Agriculture. Graduate School of Agricultural Science. Tohoku University. Japan.; (3)PhD. Agriculture. New Industry Creation Hatchery Center (NICHe). Tohoku University. Japan.

**Background and objectives:** Concentration of plasmalogen (Pls) in human plasma/serum have recently gained attention, since certain patients including those with neurodegenerative disorders have been reported to exhibit reduced levels of Pls. The possible cause of this reduction is the decline in synthesis activity and/or degradation of Pls, although it is still unknown due to the lack of a suitable analytical method. If we can characterize Pls together with its synthesis intermediates (Alkyl-phospholipid (Alkyl-PL), Lyso Alkyl-PL) and degradation products (Lyso Pls and Lyso Acyl-PL) in vivo, the cause of Pls reduction will be clarified. Therefore, in this study, we aimed to develop an accurate method to simultaneously quantitate Pls molecular species along with its synthesis intermediates and degradation products.

**Methods:** The standards (Pls, Diacyl-PL, Alkyl-PL, Lyso Alkyl-PL, Lyso Pls and Lyso Acyl-PL; Avanti Polar Lipid, Alabaster, AL, USA) were analyzed by mass spectrometry (MS/MS) in the presence of the sodium ion. The resultant product ion specific to each compound was used for the high performance liquid chromatography (LC-)MS/MS operated in multiple reaction monitoring mode. To quantitate the analytes in human plasma, analytical concerns including the extraction and matrix effect were evaluated.

**Results:** First, by the MS/MS analysis it was shown that Pls yielded a specific product ion in the presence of sodium ion. Similarly, each characteristic product ion from Diacyl-PL, Alkyl-PL, Lyso Alkyl-PL, Lyso Pls and Lyso Acyl-PL species was determined. Under the optimal LC-MS/MS conditions and with the use of the characteristic product ions, all analytes were detected with high sensitivity at femtomole levels. The extraction recoveries of all analytes from plasma were nearly 100%, and matrix effects were not observed. Moreover, as the intra-/inter-day precision and accuracy were sufficient (<20%), the present LC-MS/MS method enabled to accurately quantitate Pls, its synthesis intermediates and degradation products. We are currently applying the novel method to analyze clinical samples.

**Conclusions:** The herein developed method appears to be a powerful tool for analyzing Pls and may provide a better understanding of their physiological roles in vivo.

**Keywords:** Plasmalogen, Alkyl-acyl-phospholipid, Diacyl-phospholipid, Lyso phospholipid, LC-MS/MS.
# MATÉ TEA AND LIPID PROFILE IN OVERWEIGHT WOMEN UNDER CALORIC RESTRICTION

(1)Messina, Diego; (1)Avena, Virginia; (1)Kemnitz, Carolina; (2)Corte, Carla; (2)Mussi, Jessica; (2)Perez Elizalde, Rafael.

(1)Nutritionist. Laboratorio de Enfermedades Metabólicas. Universidad Juan Agustín Maza. Argentina.; (2)Biochemist. Laboratorio de Enfermedades Metabólicas. Universidad Juan Agustín Maza. Argentina.

**Background and objectives:** Maté tea is a traditional beverage consumed in South America, made from roasted leaves of Ilex paraguariensis. Several studies have shown its hipolipemic properties, due to the presence of polyphenols and saponins. The objective of this research was to analize the lipid profile in overweight women with or without maté consumption and or caloric restriction.

**Methods:** 119 overweight women between 25 and 50 years old were included into three groups: maté and diet (MD), maté and no diet (M), water and diet (WD). "Maté" groups drank that beverage prepared with 100 g of yerba maté on a daily basis during 12 weeks, while "diet" groups followed a low-calorie diet during that time. Anthropometric measurements and blood tests (including lipid profile: Total Cholesterol, LDL Cholesterol, HDL Cholesterol and Triglycerides) were performed at the beginning and after 12 weeks. Statistical analysis was performed by Student t-test or Wilcoxon test for paired samples and ANOVA (p<0,05 in all cases).

**Results:** After 12 weeks of treatment, Total Cholesterol fell in all groups (10,21 mg/dl in MD, 18,29 mg/dl in M, 17,63 mg/dl in W, no differences between groups). LDL Cholesterol fell in both maté groups (8,07 mg/dl in MD, 16,04 mg/dl in M group, no difference between groups); HDL Cholesterol lowered in M group (2,09 mg/dl). On the other hand, Triglycerides fell 10,74 mg/dl only in MD group.

**Conclusions:** Daily maté tea intake helps lowering Total Cholesterol and LDL Cholesterol, and provides a Triglycerid reduction together with a low-calorie diet.

**Keywords:** Ilex paraguariensis, cholesterol, dyslipidemia.

**Further collaborators.**

Laboratorio de Enfermedades Metabólicas (Universidad Juan Agustín Maza) and Instituto Nacional de la Yerba Mate.

# IRON AND ZINC CONCENTRATION IN WHEAT CULTIVARS FROM THE SOUTH BRAZILIAN RECOMMENDED LIST

(1)Scheeren, Pedro Luiz; (2)Carvalho, José Luiz Viana de; (2)Nutti, Marilia R; (2)Caierão, Eduardo; (3)Castro, Ricardo Lima de; (2)Silva, Marcio Só e; (2)Miranda, Martha Zavariz de.

(1)PhD. Geneticist and plant breeder. Embrapa Trigo. Embrapa. Brazil; (2)MSc; (3)PhD.

**Background and objectives:** Iron and zinc are important to human nutrition and for human health. The FAO Harvest Plus program is working on biofortification of human food with the objective to obtain biofortified agricultural products. In Brazil, the Embrapa's wheat breeding program begun the research of wheat biofortification in 2009. Since this time many genotypes were characterized for iron and zinc concentration in the grains. The previously results showed that levels above 40 ppm of iron and zinc are high for the evaluated material. On the other side, it is also important to know what we are consuming in our meals. So, the main objective of this research is to evaluate iron and zinc concentration in cultivars from the South Brazilian wheat recommended list.

**Methods:** Twenty nine cultivars were sowed in the field, in 2015, at the Embrapa's experimental Station. At maturity, spikes of each cultivar were harvested and threshed manually, to avoid iron and zinc contamination. The iron and zinc quantification was performed in an inductively coupled plasma optical emission spectrometer.

**Results:** For zinc, the results showed an average of 31,04 ppm of zinc. Twelve cultivars with similar concentration formed the highest group, including Vaqueano (the highest with 36,64 ppm of iron), TBIO Tibagi, BRS Parrudo, BRS Guamirim, BRS 327, Marfim, Estrela Atria, Jadeíte, FCEP 50, CD 1440, BRS Marcante and FCEP Campo Real. The lowest group was formed by TEC 10, Ametista, TBIO Sintonia, ORS Vintecinco and LG Prisma (which was the lowest with 23,82 ppm of zinc). For iron, the results showed an average of 35,10 ppm. The cultivar BRS Parrudo was superior in iron to all other cultivars, with 44,01 ppm, confirming previous evaluation. A second group was formed, in which iron concentration varied from 38,91 to 34,49 ppm. The lower group included TEC 10, BRS Marcante, TEC Frontale, Celebra, Estrela Atria, Ametista and ORS Vintecinco (with only 28,91 ppm of Iron).

**Conclusions:** After this results, BRS Parrudo showed the highest level of iron concentration in the grains and will be the basic material for continuing the breeding research for wheat biofortification in South Brazilian Region.

**Keywords:** Triticum, biofortification, cereal, breeding.
# POTENTIAL OF THE POLYPHENOL, CYANIDIN 3-O-GLUCOSIDE, IN PREVENTING CARDIOVASCULAR DEFECTS IN AN ANIMAL MODEL OF HYPERTENSIVE HEART DISEASE

(1)Netticadan, Thomas; (2)Aloud, Basma; (3)Raj, Pema; (4)McCallum, Jason; (4)Kirby, Christopher; (5)Louis, Xavier; (6)Duhamel, Todd;

(6)Wigle, Jeffrey; (1)Blewett, Heather; (4)Netticadan, Thomas.

(1)PhD. Research Scientist. Agriculture and Agri-Food Canada;

(2)MSc. PhD Student. MSc. PhD student. University of Manitoba;

(3)MSc. PhD Student. University of Manitoba. Agriculture and Agri-Food Canada; (4)PhD. Research Scientist. Agriculture and Agri-Food Canada; (5)PhD. Post Doctoral Fellow. University of Manitoba.; (6)PhD. Associate Professor. University of Manitoba.

**Background and objectives:** Polyphenols are a class of plant compounds which have gained interest in recent times due to reported health benefits. Cyanidin 3-O-glucoside (C3G) is a polyphenol found in abundance in certain dark fruits such as blackberries and black elderberries. Recent animal studies have reported that CG can prevent obesity and diabetes; however, its potential in preventing hypertension and heart disease, has not been studied. The in vitro cardiac effects of CG was studied using a cell culture model - cardiomyocytes exposed to endothelin 1 (ET1), a peptide which contributes to development of hypertension. The in vivo cardiovascular effects of C3G was examined using an animal model - the spontaneously hypertensive rat (SHR); the effects of CG was studied alone or in combination with a conventional blood pressure medication, hydrochlorothiazide (HCT).

**Methods:** For the in vitro studies, ET1 exposed cardiomyocytes and control cardiomyocytes were treated with and without CG, and cardiomyocyte hypertrophy and cell death were assessed. For the in vivo studies, five-week-old SHR and 5-week old normotensive controls [the Wistar-Kyoto rats (WKY)] were treated with and without C3G (10 mg/kg/day), HCT (10 mg/kg/day), and a combination of C3G+HCT for 15 weeks. Cardiovascular assessments included measurement of blood pressure by tail-cuff plethismography, and heart structure and function using echocardiography.

**Results:** Our in vitro studies showed that C3G prevented ET1-induced cardiomyocyte hypertrophy and cell death. Our in vivo studies showed that at 8-weeks blood pressure was lowered in SHRs treated with C3G, HCT, and C3G+HCT in comparison to SHRs. At 15 weeks SHRs treated with HCT and C3G+HCT showed lower blood pressure in comparison to SHRs, however, blood pressure was not lowered in SHRs treated with C3G alone. At 15 weeks, SHRs treated with C3G, HCT, and C3G+HCT showed a reduction in cardiac hypertrophy and improved diastolic heart function in comparison to SHRs.

**Conclusions:** The results of our study show that while CG was cardioprotective, the blood pressure lowering effect of CG was not sustained with increased severity of the disease. The results also suggest that consumption of CG rich dark berries may render cardiovascular benefits.

**Keywords:** Heart disease, polyphenol,cyanidin 3-O-glucoside, hydrochlorothiazide.

**Further collaborators:** Liping Yu MD. Research Technician. Agriculture and Agri-Food Canada.

Fahmida Jahan MSc. University of Manitoba.

# IMPACT OF OMEGA-3 FATTY ACIDS IN QUANTITATIVE AND QUALITATIVE FEATURES OF LIPOPROTEIN OF BRAZILIAN SMOKERS AND NON-SMOKERS

(1)Damasceno, Nagila; (2)Damasceno, Nagila.

(1)Department of Nutrition. Faculty of Public Health. University of Sao Paulo; (2)MSc.PhD. Department of Nutrition. Faculty of Public Health. University of Sao Paulo. Brazil.

**Background and objectives:** Cigarette smoking is an independent risk factor for cardiovascular diseases. In addition, smokers have lower plasma omega-3, which contribute to increase the cardiovascular risk in these subjects. Objectives: To evaluate the impact of omega-3 supplementation on traditional lipid profile of smokers and its relation with qualitative features of lipoproteins.

**Methods:** Based in a randomized and parallel clinical study smokers and non-smokers subjects were enrolled. Omega-3 was administrated in capsules (3.0 g/day) and all subjects were monitored during 8 weeks. After complete demographic (sex, age), clinic (medication, actual disease, familial history of disease), diet (R24h), anthropometric (weight, waist circumference) and habitual physical activity evaluation, blood samples were collected after 12h of fasting. Lipid profile (total cholesterol, LDL-c, HDL-c, triglycerides, Apo AI and Apo B), glucose, insulin, non-esterified fatty acids, oxidized LDL and subfractions of HDL and LDL were analyzed.

**Results:** Both, smoker (n=17) and non-smoker (n=44) groups showed similar age, ethnicity, diet, physical activity, current medication and Framingham risk score, however smokers group had more male (p=0.039). After 8 weeks of intervention, total cholesterol and LDL-c were reduced in smokers. Triglyceride levels decreased in both groups (smoker= -15%; non-smoker = -17%, p= 0.536) during follow-up time. There was not significant changes in LDL subfraction, however, non-smokers showed higher large HDL (smoker= +5%; non-smoker = +14%, p= 0.010) and lower small HDL (smoker= -3%; non-smoker = -16%, p= 0.024). The omega-3 was associated with non-esterified fatty acids (r=-0.512; p=0.042) and electronegative LDL (r=0.793; p<0.001) in smoker group. Smoker showed 4.85 odds (CI=1.08-21.6; p=0.039) to have the sum EPA+ DHA lower than 8% - a cut off point for high cardiovascular risk.

**Conclusions:** Omega-3 promoted positive changes in lipid profile of smokers, but only in non-smokers the qualitative features of HDL were improved.

**Keywords:** Omega-3, lipoproteins, smoking.

**Further collaborator:** Financial support: FAPESP, INCT-FCx, NAP-FCx, CNPq, CAPES.
# EFFECTS OF HONEY VARIETIES ON GLYCEMIC RESPONSES, SALIVARY GHRELIN, LEPTIN AND APPETITE RATINGS. A RANDOMIZED CLINICAL TRIAL IN HEALTHY HUMANS

(1)Emilia Aimilia Papakonstantinou; (2)Gourdomichali, Theodora;

(2)Kardara, Ioanna.

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

**Background and objectives:** Diets low in GI/GL have been associated with lower hunger, increased satiety and lower risk of chronic diseases. Glycemic index (GI) and glycemic load (GL) of honey varieties vary significantly worldwide; whereas GI/GL of Greek honey varieties is unknown. Therefore, the aims of two-studies were to: 1. determine the GI/GL of three honey varieties (fir, heather, pine) and 2. investigate their effects on salivary ghrelin, leptin, insulin and subjective appetite ratings.

**Methods:** Eleven healthy, normal-weight volunteers participated in GI investigation following International Standard Organization's protocol. Then, eleven healthy, normal-weight women consumed, cross-over, in random order, three-honey varieties, with 3-days wash-out period. Capillary blood glucose samples and salivary insulin were collected at baseline, 60 and 120min. Salivary ghrelin and leptin were collected at baseline and 120min. Subjective appetite (hunger, satiety, desire to eat) were rated on visual analogue scales (100mm, VAS) at baseline and 120min.

**Results:** Fir was medium GI (63 on glucose scale), low GL (8); heather was high GI (73), low GL (9); and pine high GI (100), medium GL (12). There was a significant main effect of time on blood glucose, salivary insulin, salivary ghrelin, subjective hunger and satiety (p for all<0.05). Peak rise in glucose was significantly higher after pine honey vs. fir and heather (p<0.001 for both). Salivary ghrelin concentrations decreased significantly after consumption of all honeys (p<0.001). Subjective hunger increased significantly after consuming fir (p=0.03) and pine (p=0.04), but not after heather (p=0.08); without significant differences between them. Subjective satiety decreased significantly after consuming fir (p=0.03) and pine (p=0.04), but not after heather; without significant differences between them. No differences were observed for fasting glucose, iAUC for blood glucose, fasting insulin, iAUC for insulin, fasting leptin, 120min-leptin and subjective desire to eat.

**Conclusions:** Honey varieties produced different GI/GL. All honeys, independently of their GI/GL, decreased salivary ghrelin concentrations, without affecting leptin or insulin, indicating decreased hunger. However, subjective hunger was increased and satiety decreased after consumption of all honey varieties. More studies are needed to investigate the effects of honey varieties on the complex issue of appetite.

**Keywords:** Honey, glycemic index, ghrelin, leptin.

# EVALUATING THE EFFECT OF DOUBLE FORTIFIED SALT ON MULTIPLE MARKERS OF IRON STATUS IN CHILDREN AND ADULTS: A SYSTEMATIC REVIEW AND META-ANALYSIS

(1)Baxter, Jo-Anna; (2)Kamali, Mahdis; (3)Gaffey, Michelle; (4)Bhutta, Zulfiqar.

(1)PhD Candidate. Centre for Global Child Health and Department of Nutritional Sciences. Hospital for Sick Children and University of Toronto. Canada.; (2)Research Assistant. Centre for Global Child Health. Hospital for Sick Children. Canada; (3)Project Manager. Centre for Global Child Health. Hospital for Sick Children. Canada.; (4)Senior Scientist. Centre for Global Child Health and Centre for Excellence in Women and Child Health. Hospital for Sick Children and Aga Khan University. Canada.

**Background and objectives:** Salt is one of few universally consumed food vehicles, making it a good candidate for fortification. Iodized salt has been highly effective in reducing iodine deficiency, thus further fortifying salt with iron could improve iron status within populations with poor access to iron supplements and widespread iron deficiency anemia. Intended to provide approximately 30% of one's daily dietary iron requirement, double fortified salt (DFS) has been developed.

**Methods:** To understand the effect of DFS compared to iodized salt (IS) on several markers of iron status, we conducted a systematic review of published and grey literature with no age restrictions. A total of 14 articles and reports were identified to meet the inclusion criteria. Meta-analyses were performed, and results were presented as mean differences (MD), standard mean differences (SMD), or risk ratios (RR) with 95% confidence intervals (CI).

**Results:** DFS was found to increase hemoglobin concentration in most studies, and the total MD in hemoglobin concentration was 3.4 g/dL (95% CI: 2.4 to 4.4; 12 trials; 39,682 individuals; I2: 95%). Anemia also significantly decreased among those provided with DFS compared to IS (RR: 0.79; 95% CI: 0.69 to 0.89; 9 trials; 36662 individuals; I2: 98%). However, no significant difference was observed between groups for serum ferritin (SMD: -0.08 µg/L; 95% CI: -1.06 to 0.91; 5 trials; 1775 individuals; I2: 99%) or transferrin receptor concentration (MD: -4.70 µg/L; 95% CI: -11.67 to 2.27; 5 trials; 1,261 individuals; I2: 98%). Notably, for all comparisons high heterogeneity was observed.

**Conclusions:** DFS could offer a modest increase in hemoglobin concentration, although its ability to improve the iron status of those at the greatest risk of poor iron-related health outcomes could be limited (i.e., pregnant women and young children). Furthermore, effective delivery of DFS could be limited by supply chain requirements.

**Keywords:** Salt, iron, iodine, food fortification.
# EFFECTS OF CONJUGATED A-LINOLENIC ACIDS (CLNAS) FROM POMEGRANATE AND BITTER GOURD SEED OILS ON INFLAMMATORY PARMETERS IN RAW 264.7 CELL CULTURES

(1)Gasparotto Sattler, José Augusto; (2)Yoshime, Luciana; (2)Melo, Illana Louise; (3)Carvalho, Eliane; (4)Mancini-Filho, Jorge.

(1)PhD Candidate. Nutricionist. Faculty of Pharmaceutical Sciences. University of Sao Paulo. Brazil.; (2)PhD. Nutricionist. Faculty of Pharmaceutical Sciences. University of Sao Paulo. Brazil.; (3)PhD. Veterinarian. Faculty of Pharmaceutical Sciences. University of Sao Paulo. Brazil.; (4)PhD. Senior Professor. Faculty of Pharmaceutical Sciences. University of Sao Paulo. Brazil.

**Background and objectives:** Punicic acid (C18:3 – 9c11t13c) and α-eleostearic acid (C18:3 – 9c11t13t), two conjugated isomers of α-linolenic acid (C18:3 – 9c12c15c), are found at high concentrations in pomegranate seed oil (PSO) and bitter gourd seed oil (BSO), respectively. Both conjugated seed oils exert anti-inflammatory effects possibly through mechanisms involving alterations in eicosanoid synthesis, reduction in the levels of signaling proteins and NF-κB, activation of PPARγ, and reduction in the levels of proinflammatory cytokines. This study aimed to investigate the anti-inflammatory effects of PSO and BSO, compared with those of linseed oil (LSO), on the activity of four cytokines, MCP-1, IL-6, IL-1β, and TNF-α, in RAW 264.7 cell cultures.

**Methods:** RAW 264.7 (monocyte/macrophage) cells were plated at a density of 5x10⁵ cells/mL and treated with samples of PSO, BSO and LSO at 5, 25 and 50 µM for 24 hours before the assessment of activities of MCP-1, TNF-α, IL-6, and IL-1β.

**Results:** Treatment with PSO, BSO, and LSO resulted in lower activities of MCP-1 and IL-6 in cells compared with those in lipopolysaccharide-stimulated cells (LPS). The activity of IL-1β showed no significant difference among LPS-treated cells and non-treated cells. TNF-α activity dropped in cells treated with BSO, PSO, and LSO.

**Conclusions:** These results can help to better understand the physiological role of CLNAs in inflammatory processes and contribute to research into the development of nutraceuticals and functional foods rich in CLNAs.

**Keywords:** Conjugated fatty acids, punicic acid, alpha-eleostearic acid, inflammatory; seed oil.

# DETERMINATION OF PARTIAL CONTENT OF PHENOLIC CONTENT AND ANTIOXIDANT ACTIVITY IN CHIA SEEDS (SALVIA HISPANICA L.)

(1)Gonzalez, Blanca Edelia; (2)López Hernández, Abad Arturo;

(3)Ortega Villarreal, Ana Sofía; (4)González Trejo, Ariadna Berenice.

(1)Universidad Autónoma de Nuevo León. Facultad de Salud Pública y Nutrición; (2)Lic. Nut. Food Laboratory. Center of Research on Nutrition and Public Health. Faculty of Public Health and Nutrition. Autonomus University of Nuevo Leon. Mexico.;

(3)MC. Food Laboratory. Center of Research on Nutrition and Public Health. Faculty of Public Health and Nutrition. Autonomus University of Nuevo Leon. Mexico.; (4)Biol. Food Laboratory. Center of Research on Nutrition and Public Health. Faculty of Public Health and Nutrition. Autonomus University of Nuevo Leon. Mexico.

**Background and objectives:** Due to the growing interest for the world's public health, functional foods and nutraceutical demand has increase. Chia seeds (Salvia hispanica L.) are a popular food of Mexico since pre-Hispanic, it's considered as a functional food due to its bioactive compounds believed to have health benefits. It possesses a high content of phenolic compounds, which are important for the protection of the lipid oxidation that impact as much as the health of the users as food quality.

**Objectives:** Determine the content of phenolic compounds and antioxidant activity in methanolic extract of chia seeds.

**Methods:** The chia seeds used in this project were cultivated in Mexico. Extracts were made with 50g of chia in 250ml of methanol-maceration concentrated by rotavaporation.

Antioxidant capacity was determined using DPPH {2,2-diphenyl-1-picrylhydrazyl} (Bondet, 1997), FRAP {ferric reducing antioxidant power} (Benzie y Strain, 1996) and ABTS {2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)} (van den Berg, 2000) methods, expressing results as mM of TEAC g-1 {mmol Trolox equivalent antioxidant capacity/g}. Previous to the quantification of phenolic compounds by HPLC {High Performance Liquid Chromatography} an hydrolytic split off of the glycosides of polyphenols was performed following the methodology proposed by Taga et al., (1984) and Hempel and Bohm (1996) for the chromatographic technique, with modifications by analyzing the extracts before and after hydrolysis. Total polyphenol content was determined as mg GAE/g {milligrams equivalent of gallic acid per gram of extract} by the Folin-Ciocalteu method proposed by Singleton and Rossi 1965.

**Results:** The antioxidant activity of the extracts was 2.49±0.15, 8.32±0.91 and 4.77±0.38 mM TEAC g-1 for DPPH, FRAP and ABTS respectively. Total polyphenol content was 1.08 ± 0.03 mg GAE/g. In the chromatographic analysis, caffeic acid was found mainly in the non-hydrolyzed extract (3.47±0.59 mg/g) followed by chlorogenic acid (0.51±0.16 mg/g). In non-hydrolyzed extracts, after hydrolysis was found: myricetin 1.21±0.89 mg/g, quercetin 2.70±1.14 mg/g and kaempferol 0.27±0.12 mg/g.

**Conclusions:** Regarding the above, chia seeds can be considered as a good component of human diet, due to its high phenol-
ic compounds content (caffeic acid, chlorogenic acid, myricetin, kaempferol and quercetin) with antioxidant activity, which can lead to health benefits.

**Keywords:** Chia seeds, antioxidant activity, phenolic compounds.

144/2183

EFFECT OF THE STIMULATION WITH (-) EPICAT-
ECHIN ON GENE EXPRESSION IN HUVEC CELLS

(1)Tejero, Maria Elizabeth; (2)Suizer-Ponce de León, Sandra; (3) Barrera-Reyes, Paloma; (4)Ruiz-Leyja, Estela D´Abril; (5)Vadillo-Ortega, Felipe.

(1)PhD. Researcher. Laboratorio de Nutrigenomica. Instituto Nacional de Medicina Genomica. Mexico.; (2)Nutrition Student. Universidad Iberoamericana. Mexico; (3)PhD student. Doctorado en Ciencias Biomédicas. UNAM. Mexico; (4)M Sc student. Maestria en Biologia Experimental. UAM-I. Mexico; (5)MD. PhD. Facultad de Medicina. UNAM. Mexico.

**Background and objectives:** Epicatechin (EC) is the most abundant polyphenol in cocoa. Consumption of cocoa polyphenols has been related with cardiovascular benefits such as decrease in blood pressure and improvement of markers of glucose and lipid metabolism in human. These polyphenols have been shown to modulate gene expression and exert an anti-inflammatory effect. The present study analyzed the dose-response effect of EC stimulation on the expression of genes related with inflammation (NFkB) and endothelial cell function (eNOS) in primary culture of human umbilical vein endothelial cells (HUVEC). The tested doses included concentrations that are achievable by intake of EC containing food.

**Methods:** HUVEC cells were isolated from the umbilical cord of women with a history of healthy pregnancy who signed the informed consent. Endothelial cells were isolated and cultured using standard methods. Cells were stimulated with three different doses of EC (1, 0.1, 0.01 uM and untreated control) during 4 h. All experiments were conducted in triplicates. RNA was isolated from cells and eNOS and NFkB (p50) mRNA was measured using real time, quantitative RT-PCR by absolute quantitation. HUVEC cells were identified by immunofluorescence staining. Comparison of the mRNA abundance across treatments was conducted by a one-way ANOVA. Statistical analysis was conducted in SPSS v.20 software. This project was approved by the Ethics committee at INMEGEN.

**Results:** The immunofluorescence staining was positive for endothelial cells. Significant differences in mRNA abundance were found for the analyzed genes. Stimulation with EC at 0.01 uM decreased the expression of eNOS mRNA as compared with control (p<0.001). The expression of NFkB significantly increased after the stimulation with 0.1 and 0.01 uM of EC as compared with control. No effect of the highest dose was observed on the analyzed transcripts.

**Conclusions:** These results suggest that treatments induced moderate pro-inflammatory changes in the analyzed genes. Further investigation is required to determine the effect of EC at different times and changes in the protein products of the analyzed genes.

**Keywords:** Epicatechin, eNOS, NFkB, endothelial cells, cocoa.

144/2201

EXPRESSION MODULATION OF AGTR1A AND
BDKRB2 GENES IN HYPERTENSIVE RATS TREAT-
ED WITH EXTRACT OF CHIA SEEDS

(1)Gonzalez, Blanca Edelia; (2)Arredondo Mendoza, Gerardo Ismael;

(3)Guzmán de la Garza, Francisco Javier; (4)Campos Góngora, Eduardo.

(1)Universidad Autónoma de Nuevo León. Facultad de Salud Pública y Nutrición; (2)Lic. Nut. Investigation Centre of Public Health and Nutrition. Autonomous University of Nuevo León. Mexico.; (3)Dr. en C. Investigation Centre of Public Health and Nutrition. Autonomous University of Nuevo León. Mexico;

(4)Dr. en C. Investigation Centre of Public Health and Nutrition. Autonomous University of Nuevo León. Mexico.

**Background and objectives:** High Blood Pressure (HBP) is the principal risk factor for cardiovascular disease. HBP is regulated by the renin-angiotensin system, particularly the angiotensin-converting enzyme (ACE). ACE inhibitors are used as first-line therapy, however, these produce secondary reactions. Searching of natural source is an alternative for HBP treatment and salvia species have shown a hypotensive effect. We analyzed the effect of an ethanolic extract of chia (Salvia hispanica L.) seeds (EESH) on the expression of Agtr1a and Bdkrb2 genes, involved in HBP regulation.

**Methods:** A hypertension rat model was generated by administration of a nitric oxide synthesis inhibitor: NG-nitro-L-arginine methyl ester (L-NAME). Hypertensive rats (12) were allocated to three different treatments; the first group received captopril, the second group received EESH and the last group remained with administration of L-NAME. Control group (healthy group, 4 rats) was treated with distilled water during the whole study. In the second phase, gene expression analysis was realized by RT-PCR in cardiac tissue samples, using specific oligonucleotides designed for Agtr1a, Bdkrb2 and Gapdh genes. Comparisons of level expression between groups was performed using statistical analyses with ANOVA test.

**Results:** Rats treated with L-NAME increased its blood pressure: between 310 to 328 mmHg, while the control group showed values of 100 mmHg. After, the administration of the different treatments, rats that was receiving L-NAME showed an increment on blood pressure levels (31.2%), and the administration of captopril and EESH decreased 0.2% of blood pressure levels. Gene expression analysis showed that the administration of L-NAME incremented significantly the expression of Agtr1a gene, in comparison to basal levels (healthy rats). In the hypertensive rats treated with captopril there were no changes in the expression of Agtr1a. The administration of EESH produced a similar effect as captopril in Agtr1a expression levels. The Bdkrb2 gene expression did not show significative changes in the different treatment groups.

**Conclusions:** The EESH shown to have a similar effect to the captopril on the blood pressure of hypertensive rats. Besides, a regulatory effect of the expression of the genes involved on the modulation of blood pressure was observed.

**Keywords:** High Blood Pressure, Salvia hispanica L, expression genes.
PHYSICOCHEMICAL, FUNCTIONAL AND NUTRITIONAL CHARACTERIZATION OF RAW FLOUR OBTAINED FROM DIFFERENT SWEET POTATO (IPOMOEA BATATAS) AND ÑAMPI (DIOSCOREA TRIFIDA)

(¹)Quintero Montenegro, Rosa Itzela; (²)Valenzuela, Ximena;

(³)Sevillano, Alejandrino; (⁴)Rivera, Dayra.

(¹)Dra. Ciencias y Tecnología de Alimento. Universidad Tecnológica de Panamá; (²)MSc Laboratorio de Envase de la Universidad de Santiago de Chile; (³)Licdo. Ciencias y Tecnología Innovadora de Alimentos. Universidad Tecnológica de Panamá;

(⁴)Licda. Ciencias y Tecnología Innovadora de Alimentos. Universidad Tecnológica de Panamá.

**Background and objectives:** The main objective in food processing is the use of inexpensive raw materials, high durability and high amount of compounds with nutritional value, in order to reduce the potential costs and prevent deterioration, establishing as a priority the search for new alternatives to increase its conservation.

The use of technological purpose flour in bread and pastry requires knowledge of the characteristics regarding the ability to develop along with the other ingredients that make up the mass. These characteristics of the flour contemplated parameters humidity, water absorption, and enzyme activity bread making quality. This paper aims to characterize two types of flour from artisanal mining, in order to determine their physical, nutritional, functional characteristics and potential uses in baking and pastry

**Methods:** For this performance from the fruit from a craft collection process it was determined by a proximal composition and activity antioxidants was determine by method DPPH and polyphenols totals. Also, determining the thermal qualities thermal analysis (DSC) and thermo gravimeter (TGA) was performed

**Results:** Statistically significant differences were found between the samples flour studied humidity (1.88 – 3.57%), ashes (3.51 – 4.42%), protein (5.74 ± 1.36 %), crude fat (0.262 - 1.52%), total carbohydrate (4.48 – 9.02%) and polyphenols (200.8 ± 50.67 mg GAE/100 g dry mass)

The swelling and solubility of both flours showed similar changing patterns as the temperature increased.

**Conclusions:** The flour obtained show physicochemical characteristic and chemical composition desirable mainly by high protein contain, total carbohydrates. Also, presented high capacity water absorption and swelling power. On the other hand, the phenol content and antioxidant capacity were

All the flour has excellent potential as ingredients for new baking product development

**Keywords:** Flour, sweet potato, ñampi, tropical roots, functional properties.

**Further collaborators.**

Dr. Abel Guarda Moraga

CONSUMPTION OF MILK PRODUCTS WITH 100% B-CASEIN A2 IMPROVES OVERALL GASTROINTESTINAL TOLERANCE BUT HAD NO EFFECT ON BEHAVIOR OF MEXICAN CHILDREN WITH AUTISM SPECTRUM DISORDER

(¹)Pedroza García, Karina Alejandra; (¹)Ronquillo, Dolores; (²)Palacios Delgado, Jorge Raúl; (²)Anaya-Loyola, Miriam Aracely; (²)Rosado, Jorge L.

(¹)MD. Facultad de Ciencias Naturales. Universidad Autónoma de Querétaro. México; (²)PhD. Facultad de Ciencias Naturales. Universidad Autónoma de Querétaro. México.

**Background and objectives:** Autism spectrum disorders (ASD) affect two areas of development, the first comprises restricted, repetitive and stereotyped behaviors, second refers to alterations in social communication. There is evidence of behavioral improvement of children with ASD following casein free diets because digestion of β-casein A1, but not β-casein A2, produce an opioid peptide called β-casomofphin 7 which has been associated with behavioral disturbances and gastrointestinal problems. The aim of the study was to compare the effect of milk and cheese ingestion with either a combination of A1/A2 or only A2 β-casein on ASD children's behavior, milk tolerance and intestinal permeability.

**Methods:** A double-blind, randomized 16-week crossover study was conducted. 22 ASD children and adolescents participated. Subjects underwent 2-week dairy washout followed by 6 weeks of dairy products with β-casein A2 or A1/A2 and a further 2-weeks dairy washout followed by final 6 weeks of dairy products with β-casein A2 or A1/A2. Child Behavior Checklist (CBCL) and Repetitive Behavior Scale-Revised (RBS-R) were applied to measure behavioral changes in internalized and externalized behaviors and repetitive behaviors. Intestinal permeability was evaluated by a lactulose/mannitol test in urine by HPLC. Gastrointestinal tolerance was measured by questionnaire in which the number of bowel movements, consistency, diarrhea, constipation, or loss of appetite were recorded.

**Results:** No changes were observed in the psychometric questionnaires RBS-R (0.6±8.7 vs. -5.6±9.3, p=0.06) and CBCL (12.8±2.7 vs.13.6±2.7, p=0.16) between treatments which could be a reflection of the integrity of the intestinal permeability, average L/M was 0.01 ± 0.02. Gastrointestinal tolerance was consistently improved during the intake of products with β-casein A2 than β-casein A1/A2.

**Conclusions:** In this initial study we conclude that the intake of milk and milk products with 100% of β-casein A2 have benefits in gastrointestinal tolerance but not beneficial effects in overall behaviour of children with ASD. Since mucosal permeability was not affected in our children, we cannot rule out the possibility that milk with 100% of β-casein A2 has a positive effect on the behavior of children with ASD who have increased intestinal permeability or frequent gastrointestinal problems.

**Keywords:** Autism spectrum disorder, behavior, β-casein A2, intestinal permeability, milk tolerance.

**Conflict of Interest Disclosure:** Jorge L Rosado has stocks in a Company that produces milk with β-casein A2, but he did not intervene in data analysis. The remaining authors declare no conflict of interest.
# TOTAL POLYPHENOL CONTENT AND ANTIOXIDANT CAPACITY IN JUICE AND PEEL OF BLUEBERRIES (VACCINIUM CORYMBOSUM) GROWN IN CAÑETE VALLEY, PERU

(1)Lon Kan Prado, Elena Elizabeth; (2)Macavilca Ticlayauri, Edwin Antonio; (3)Lon Kan Prado, Carlos Alberto; (4)Canchos Chipana, Carlos; (5)Reyes Veramendi, Jyothisa Eva-Gina.

(1)PhD. Professor Nutritionist. School of Nutrition. Universidad Le Cordon Bleu. Peru.; (2)Engineer. Professor. School of Food Engineering. Universidad José Faustino Sanchez Carrion. Peru.;

(3)MSc. Professor. School of Nutrition. Universidad Le Cordon Bleu. Peru.; (4)BSc. Food Engineering. School of Food Engineering. Universidad José Faustino Sanchez Carrion. Peru.

**Background and objectives:** Blueberries are considered as the king of antioxidant foods, due its high content that protects our bodies from damage by free radicals, unstable molecules that can damage cellular structures and contribute to aging and diseases like cancer. The aim of the present study was to determine the total polyphenol content and antioxidant capacity of juice and peel from blueberries (Vaccinium corymbosum) grown in Cañete Valley, Peru.

**Methods:** Folin-Ciocalteu and DPPH methods were used to determine the concentration of total polyphenol content and antioxidant activity, respectively. Extracts were obtained from juice and peel in four study parameters. At first, solvents with different polarities were tried (water, methanol, ethanol 80% and acetone-water 75%); second, ratio fruit/solvent; thirth, extraction-time and fourth, temperature.

**Results:** Results in juice: total polyphenol content 3,474 mg Gallic acid/ml juice and antioxidant capacity 12,131 Trolox mg/ml juice. Results in peel: showed solvent acetone/water 75% > ethanol/agua 80% > methanol > distilled water. The ratio 1/10 was greater ratio 1/12. 4 hours-time exhibited the higher values in total polyphenol content 16.40 mg Gallic acid/ml extract and antioxidant capacity 8,90 mg Trolox/g blueberries. The effect of temperatures were studied between 40 to 80 °C. At 60, 70 and 80 °C showed higher total polyphenol content values: 19,213; 26,522; 32,830 mg Gallic acid/ml extract, respectively; however the stability and antioxidant capacity were reduced when increased the temperature: 13,500; 13,675 and 12.525 mg Trolox/g peel, respectively; and at temperatures below about 40-50 °C the total polyphenol content and the antioxidant capacity showed higher stability. It is concluded that the peel has higher total polyphenol content and antioxidant capacity than the juice and fresh whole fruit.

**Conclusions:** The stability and antioxidant capacity were reduced when increased the temperature (13,500; 13,675 and 12.525 mg Trolox/g peel, respectively).

At temperatures below about 40-50 °C the total polyphenol content and the antioxidant capacity showed higher stability.

It is concluded that the peel has higher total polyphenol content and antioxidant capacity than the juice and fresh whole fruit.

**Keywords:** Blueberries, polyphenol content, antioxidants capacity.

# EFFECT OF DEEP FRYING ON BIOACTIVE COMPONENTS OF WHEAT FLOUR PRODUCTS

(1)Prakash, Jamuna; (2)Oghbaei, Morteza.

(1)Professor (Retd.), Department of Food Science and Nutrition, University of Mysore, India.; (2)Research Scholar.

**Background and objectives:** Deep frying is one of the most common cooking methods used for many products, and in particular for cereal based snacks and dishes. Frying uses high temperature and could lead to partial destruction of bioactive components. Frying also changes the microstructure of foods altering the bioavailability of its constituents.

**Objective:** Hence the present investigation was undertaken to determine the effect of deep frying wheat flour based products on their bioactive components namely, total phenols and flavonoids.

**Methods:** The flours used were whole wheat (WWF) and refined flour (RWF) as these are commonly used for preparation of staple products. Mineral fortification of flour was one of the variation chosen as many times staple flours are fortified with minerals in developing countries. Whole and refined wheat flour fortified either with iron or zinc (each with 10 & 20mg) were used for preparing 'Chapathis' (Indian unleavened pan-roasted flat bread), and 'Poories' (deep fried flat bread) and analyzed for total and in vitro bioavailable total phenols and flavonoids.

**Results:** Results showed that the total phenol content in chapathi was higher in WWF products (83.25-93.74 mg/100g) and much lesser in RWF products (19.82-25.96 mg/100g). Similar trend was seen in bioavailability pattern (60-77% for WWF and 36-40% in RWF). Wheat had a small flavonoid content which was higher in WWF products than RWF products. However bioavailability was slimier in WWF and RWF. On deep frying, there was a visible decline in the bioactive constituents, both total phenols and flavonoids reduced in fried products. The bioavailability was also much lesser in comparison with pan-roasted products. In WWF poories, 43-58% of total phenols were bioavailable whereas similar range for RWF poories were 17-21%. Again the RWF products had much lesser availability than WWF products indicating that both refining and deep frying were detrimental to content and availability of bioactive components in wheat flour.

**Conclusions:** Hence, limiting the use of refined flour as well as deep frying is beneficial to retain maximum bioactive components.

**Keywords:** Total phenols, flavonoids, bioavailability, roasting, frying.
RECOMMENDATION VS. REALITY: A GLOBAL ASSESSMENT OF FRUIT AND VEGETABLE INTAKE AND VARIETY

(1)Randolph, Keith; (2)Murphy, Mary M; (3)Barraj, Leila; (3)Spungen, Judith; (4)Herman, Dena R; (5)Randolph, Keith.

(1)PhD. Technology Strategist. Nutrilite Health Institute. United States; (2)Exponent Inc. Washington DC. USA; (3)Exponent Inc. Washington. USA.; (4)Department of Family and Consumer Sciences. California State University. Northridge. USA.; (5)PhD. Nutrilite Health Institute. USA.

**Background and objectives:** In 2004 the World Health Organization (WHO) and Food and Agricultural Organization (FAO)

established minimum fruit and vegetable intake recommendations for adults at 400 g or 5 servings per day. The basis for this recommendation was epidemiological data that showed a strong inverse relative risk for chronic disease morbidity with fruit and vegetable intakes up to this level. Despite this recommendation, there has not subsequently been a global assessment of fruit and vegetable intake.

**Methods:** Utilizing WHO World Health Survey data (2002) and FAO food supply utilization data (2012), we evaluated fruit and vegetable intake, fruit and vegetable variety, and intakes of selected phytonutrients in different regions globally.

**Results:** The results show a striking shortfall globally in fruit and vegetable intake relative to the WHO minimum recommendation, and relatively narrow range of variety. According to this assessment, approximately 75% of individuals worldwide fail to meet WHO minimum intake levels.

**Conclusions:** These data strongly support the need to better educate consumers regarding the health promoting and risk reducing effects of fruit and vegetable dietary patterns.

**Keywords:** Fruit and vegetable intake, dietary pattern.

**Further collaborators.**

Alli Klosner

EFFECT OF AQUEOUS LEAVES EXTRACT FROM PASSIFLORA ALATA CURTIS AND VITEXIN, ISORIENTIN IN CO-CULTURE OF MIN6/LYMPHOCYTES FROM NOD MICE IN OXIDATIVE STRESS AND CELL DEATH

(1)Colomeu, Talita Cristina; (2)Colomeu, Talita Cristina; (2)De Figueiredo, Daniella; (3)Silva Matos, Priscila; (3)de Campos Carvalho, Virginia; (2)Simon Gonzalez Schumacher, Nayara; (3)Moitinho Abram, Debora; (4)Romani Fernandes, Luis Gustavo; (5)de Lima Zollner, Ricardo.

(1)PhD student of medical clinical. Faculty of Medicine. University of Campinas.Brazil; (2)PhD student of clinical medical. Laboratory of Translational Immunology, Department of Internal Medicine, University of Campinas, Campinas, São Paulo, Brazil; (3)M.S.c student of clinical medical.Laboratory of Translational Immunology, Department of Internal Medicine, University of Campinas, Campinas, São Paulo, Brazil; (4)PhD. Biologist. Laboratory of Translational Immunology, Department of Internal Medicine, University of Campinas, Campinas, São Paulo, Brazil; (5)PhD. Prof. Doctor Laboratory of Translational Immunology, Department of Internal Medicine, University of Campinas, Campinas, São Paulo, Brazil.

**Background and objectives:** Type 1 Diabetes mellitus can be associated with increase of oxidative stress and infiltration of cells of immune system, such as lymphocytes CD4+ and CD8+. The Palata is considered by the Brazilian Pharmacopoeia as a medicinal plant that plays important role decreasing inflammation. The aim was to evaluate the effect of Palata aqueous leaves extract and vitexin and isoorientin in cell inhibition and free radicals production.

**Methods:** NOD mice spleen lymphocytes were isolated through the wool column/2,5x10⁵ and cultured with min6 cells/8x10⁴. The co-cultures were treated with different concentrations of P.alata and vitexin and isoorientin and analyzed in proliferation of CD4+ and CD8+ lymphocytes to evaluate the concentration of inhibition dose (ID50). The ID50 were used to analysis of Apoptosis, Mitochondria Depolarization, CellRox oxidative stress reagentes, MitoSOX red mitochondrial superoxide indicator and Glutathione ThiolTracker Violet. All analyses were done by flow cytometry. The supernatants were used to evaluate insulin production by ELISA in min6 cells.

**Results:** The concentration of inhibition proliferation of Palata was 500µg/mL, Vitexin 200µM and Isoorientin 5µM. The apoptosis demonstrated most of cells in late apoptosis/necrosis and the mitochondrial depolarization was higher in Palata and Vitexin when compared with Isoorientin. CellRox showed higher in all treatment and Glutathione decrease in all treatment, but in Palata showed greater decrease of this antioxidant. The production of superoxide treated with Vitexin and Palata was higher in CD4+. Insulin analysis showed high production of hormone in all treatment. The CD4+/CD8+ lymphocytes are the main type of cells involved in destruction of β cells during type 1 Diabetes Mellitus. The results showed that CD4+ and CD8+, when treated with Palata or Vitexin and Isoorientin, can inhibit cell prolifera-
tion through mitochondrial depolarization increase the oxidative stress and consequently cell death. Besides that, the high production of insulin demonstrated that β cell in min6 continued to produce this hormone, maintaining viability cellular.

**Conclusions:** In conclusion, the treatment with P.alata or vitexin and Isoorientin leads to lymphocytes death triggered by increased oxidative stress without causing damage to β cells, therefore these treatments can aid in decreasing cell infiltration in β cells.

**Keywords:** Passiflora alata curtis, vitexin and isoorientin, antioxidant activity, decrease inflammation.

144/2695

EFFECT OF AQUEOUS EXTRACT OF PASSIFLORA ALATA CURTIS, CATECHIN AND RUTIN IN PROLIFERATION, APOPTOSIS, ERK PHOSPHORYLATION AND AKT SIGNALING PATHWAY OF LYMPHOCYTES (NOD MICE) FROM CO-CULTURE WITH MIN6

**Background and objectives:** Passiflora alata curtis leaves (passion fruit) contains phenolic compounds in its composition with anti-inflammatory and antioxidant properties. This study aimed to investigate the effects of aqueous leaf extract of P. alata, catechin and rutin, compounds presents in the leaves, in lymphocytes submitted to co-culture with MIN6 cells.

**Methods:** Different concentrations of P. alata (50, 200 and 500µg/mL), catechin (25, 50 and 75µM) and rutin (50, 150 and 300µM) were added separately to cell culture containing lymphocytes (2.5×10⁵) from diabetic NOD mice and MIN6 cells (8×10⁴) and it was maintained in culture for 96 hours (CO₂ 5% at 37°C)

for analysis of proliferation, apoptosis, ERK phosphorylation cascade's and AKT signaling pathway in TCD4+ and TCD8+ populations. The presence of inflammatory cytokines and insulin in culture supernatant was also analyzed. All analyses were done by flow cytometer (Guava easycyte, Millipore*, Bellirica, USA).

**Results:** It was observed a decrease in the percentage of CD4+ and CD8+ T cells proliferation after each treatment. The concentrations of P. alata (500µg/mL), catechin (50µM) and rutin (300µM) to be used in apoptosis, ERK, AKT and supernatant analysis were determined calculating 50% of inhibition of lymphocytes proliferation (ID50). Increase in the percentage of lymphocytes in apoptosis and late apoptosis/necrosis was observed when submitted to P. alata. AKT analysis showed lower expression in lymphocytes submitted to P. alata and catechin while lymphocytes in the presence of P. alata and both compounds presented lower percentage of ERK phosphorylation. Decrease in TNFα and IFNγ cytokines was observed in supernatants of P. alata and rutin treatment while the expression of insulin was preserved after all treatments.

**Conclusions:** TCD4+ and TCD8+ cells develops an important role in triggering type 1 diabetes whereas these cell types attack the pancreatic islets destroying beta cells. In addition, inflammatory cytokines such as TNFα and IFNγ are important mediators of beta-cell death induced by CD4+ T cells. The decrease in the levels of these cytokines after treatment associated with preservation of insulin, inhibition of proliferative T cells, induction of apoptosis and decrease in the expression of AKT/ERK may provide useful information for the treatment of autoimmune diabetes with natural products.

**Keywords:** Passiflora alata, rutin, catechin, anti-inflammatory effect, NOD mice.

**Further collaborators:** Supported by: CNPq and FAPESP

144/2890

EFFECTS OF GENETIC AND ENVIRONMENTAL FACTORS ON PROANTHOCYANIDINS IN SEA BUCKTHORN (HIPPOPHAË RHAMNOIDES)

**Background and objectives:** Sea buckthorn (Hippophaë rhamnoides) berries are rich sources of flavonoids and other polyphenols. Proanthocyanidins (PAs) in sea buckthorn have biological activity and may be effective in maintaining health.

**Methods:** Sea buckthorn berries of three subspecies (H. rhamnoides ssp. rhamnoides, ssp. mongolica and ssp. sinensis) including nine varieties grown in Finland, Canada and China were

<footnote>(1)De Figueiredo, Daniella; (1)Figueiredo, Daniella; (1)Colomeu, Talita Cristina; (2)Silva Matos, Priscila; (3)de Campos Carvalho, Virginia;</footnote>

<footnote>(4)Simon Gonzalez Schumacher, Nayara; (2)Moitinho Abram, Debora;</footnote>

<footnote>(5)Romani Fernandes, Luis Gustavo; (6)de Lima Zollner, Ricardo.</footnote>

<footnote>(1)PhD student. Biologist. Laboratory of Translational Immunology. Department of Internal Medicine. Faculty of Medical Sciences. University of Campinas. Campinas. São Paulo. Brasil.; (2)MSc student. Biomedical. Laboratory of Translational Immunology. Department of Internal Medicine. Faculty of Medical Sciences. University of Campinas. Campinas. São Paulo. Brasil.; (3)MSc student. Biologist. Laboratory of Translational Immunology. Department of Internal Medicine. Faculty of Medical Sciences. University of Campinas. Campinas. São Paulo. Brasil.; (4)PhD student. Biomedical. Laboratory of Translational Immunology. Department of Internal Medicine. Faculty of Medical Sciences. University of Campinas. Campinas. São Paulo. Brasil.; (5)PhD. Biologist. Laboratory of Translational Immunology. Department of Internal Medicine. Faculty of Medical Sciences. University of Campinas. Campinas. São Paulo. Brasil.; (6)PhD. Laboratory of Translational Immunology. Department of Internal Medicine. Faculty of Medical Sciences. University of Campinas. Campinas. São Paulo. Brasil.</footnote>

<footnote>(1)Yang, Wei; (2)Laaksonen, Oskar; (3)Kallio, Heikki; (4)Yang, Baoru.</footnote>

<footnote>(1)Food Chemistry and Food Development. Department of Biochemistry. University of Turku. Finland; (2)PhD. Food Chemistry and Food Development. Department of Biochemistry. University of Turku. Finland; (3)PhD. Professor (emeritus). Food Chemistry and Food Development. Department of Biochemistry. University of Turku. Finland; (4)PhD. Professor. Food Chemistry and Food Development. Department of Biochemistry. University of Turku. Finland.</footnote>