# DO DIETARY PATTERNS DETERMINE LEVELS OF VITAMIN B6, FOLATE, AND VITAMIN B12 INTAKES AND CORRESPONDING BIOMARKERS IN EUROPEAN ADOLESCENTS? THE HELENA STUDY

(1)1 GENUD (Growth' Exercise' NUtrition and Development) Research group. Universidad de Zaragoza. Instituto Agroalimentario de Aragón (IA2). Instituto de Investigación Sanitaria Aragón (IIS Aragón). Zaragoza. Red de Salud Materno-infantil. Spain; (2)PhD. Nutritionist. Department of Public Health. Ghent University. Ghent. Belgium. International Agency for Research on Cancer (IARC). 150 Cours Albert Thomas. 69372. Lyon. Cedex 08. France.; (3)PhD. Nutritionist. GENUD (Growth' Exercise' NUtrition and Development) Research group. Universidad de Zaragoza. Spain; (4)PhD. Statistician. Department of Preventive Medicine and Public Health. Universidad de Zaragoza. Zaragoza. Spain.; (5)PhD. Nutritionist. Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III. Madrid. Spain. GENUD (Growth' Exercise' NUtrition and Development) Research group. Universidad de Zaragoza. Spain.; (6)PhD. Nutritionist. GENUD (Growth' Exercise' NUtrition and Development) Research group. Universidad de Zaragoza. Spain. Instituto Agroalimentario de Aragón (IA2). Instituto de Investigación Sanitaria Aragón (IIS Aragón). Zaragoza. Spain.; (7)PhD. MD. Université Lille. CHU Lille. LIRIC UMR 995 Inserm. Clinical Investigation Centre. CIC-1403-Inserm-CHU. F-59000 Lille. France.; (8)PhD. MD. Research Institute of Child Nutrition. Pediatric University Clinic. Ruhr-University Bochum. Germany.; (9)PhD. Pharmacologist. ImFINE Research Group. Department of Health and Human Performance. Universidad Politécnica de Madrid. Madrid. Spain.; (10)PhD. MD. GENUD (Growth' Exercise' NUtrition and Development) Research group. Universidad de Zaragoza. Spain. Instituto Agroalimentario de Aragón (IA2). Instituto de Investigación Sanitaria Aragón (IIS Aragón). Zaragoza. Spain.

**Background and objectives:** Adolescence is a period in which micronutrients status might be compromise due to frequent consumption of food with low nutritional value and due to increased requirements for growth and development. Among those, B-vitamins, as key-role micronutrients, might be affected. For this reason, the aim of this study is to determine the dietary patterns (DPs) explaining the highest variance of vitamin B6, folate, and B12 intake and related concentrations among European adolescents.

**Methods:** 2,173 adolescents participating in the HELENA (Healthy Lifestyle in Europe by Nutrition in Adolescence) study met the eligibility criteria for B-vitamins intake analysis (46 % males) and 586 did it for biomarkers analysis (47 % males). Two non-consecutive 24-h dietary recalls were used to assess mean

intakes. Concentrations were measured by chromatography and immunoassay. Reduced rank regression was applied to elucidate the combined effect of food intakes in B-vitamins intakes and concentrations.

**Results:** Identified dietary patterns (one per each B-vitamin intake and biomarker and by sex) explained a variability between 34.2 % and 23.7% of the B-vitamin intakes and between 17.2 % and 7% of the biomarkers. Fish, eggs, cheese, and white and buttermilk intakes, loaded positively for B-vitamins intake in both sexes; in contrast, soft drinks and chocolate, loaded negatively. For biomarkers, there was a higher heterogeneity in terms of foods loads in the identified patterns, like in the case of cheese, that loaded negatively for the identified pattern in males and females for plasma vitamin B12, and positively in the pattern of males and females for RBC-folate, and also positively for PLP in females. Besides, some food items loaded differently between intakes and biomarkers, like fish products which, in females, loaded positively for intakes, but negatively for plasma folate.

**Conclusions:** The identified dietary patterns explained more variance of the B-vitamins intake than for their related biomarkers both in males and females. The dietary patterns were different for the same B-vitamin between males and females, and between intake and status of the same B-vitamin. Further studies are needed to elucidate the factors determining such differences.

**Keywords:** Dietary patterns, Reduced Rank Regression, B-vitamins, Europe, adolescents

**Further collaborators:**

Yannis Manios, Alejandro De la O Puerta, Anthony Kafatos, Ascensión Marcos, Stefania Sette, Maria Plada, Peter Stehle, Dénes Molnár, Kurt Widhalm, Stefaan De Henauw.

# RELATIONSHIP BETWEEN NON NUTRITIVE SWEETENERS INTAKE AND NUTRITIONAL STATUS

García, María Isabel; Tello, Sofía; Poch, Clarisa; Rellecati, Candela; Muguerza, María de los Ángeles; Capaldi, Luciana; Ruiz, Laura Matilde.

LDN. Nutritionist. Department of food and nutrition. Centro Gallego de Buenos Aires. Argentina.

**Background and objectives:** Obesity and associated metabolic diseases are going through a constant worldwide increase. At the need to prevent and/or maintain a good health condition an interest for non-nutritive sweeteners (NNS) rise. Although their impact on intake and body weight is unclear, its consumption is high for being widely distributed in foods. Epidemiologic studies associate NNS consumption with excessive food intake and body weight gain.

The objectives are to estimate NNS consumption's prevalence in diet products on an adult population, evaluate association between nutritional status and NNS consumption and estimate NNS intake and compare it with the Acceptable Daily Intake (ADI).

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<footnote>(4)Santabárbara, Javier; (5)Fernández-Alvira, Juan Miguel;</footnote>

<footnote>(6)Santaliestra-Pasías, Alba M.; (7)Gottrand, Frédéric; (8)Kersting, Mathilde; (9)González-Gross, Marcela; (10)Moreno, Luis A.</footnote>
termed by real-time PCR. Also, duodenum and jejunum was
ligated and water-soluble heme or hemin iron was administered
to measure the uptake of iron into mucosal cells and portal vein
blood.

**Results:** Levels of expression of HO-1 and Fpn1 mRNA were high in the duodenum, and these were upregulated in iron-deficient rats. In the duodenum, iron uptake was about 30% after loading with water-soluble heme iron, but hemin was not absorbed. Levels of expression of Hcp1 and Flvcr1 mRNAs were high in the jejunum, and iron uptake was also high after loading with hemin in this section. Hcp1 and Flvcr1 mRNAs were not upregulated by iron deficiency.

**Conclusions:** In the duodenum and jejunum of the small intestine, different iron absorption mechanisms for water-soluble heme iron and hemin are considered to exist.

**Keywords:** heme iron absorption, HCP1, Flvcr1, iron deficiency, hemin

and fasting insulin (70.2 vs 42.4 pmol·L; p<0.05). There were not
significant differences in plasma total FFAs concentration between
groups. However, children with AO had higher palmitoleic acid
(0.94 vs 0.70 wt%; p<0.05) and dihomo-gamma linoleic acid (2.76
vs 2.07 wt%; p<0.05). Palmitoleic and dihomo-gamma linoleic ac-
ids correlated (p<0.05) with BMI (r=0,397; r=0,296, respectively)
and waist circumference (r=0,380; r=0,276, respectively). Myristic
and palmitoleic acids correlated (p<0.05) with high-density lipo-
protein cholesterol (r=-0.408; r=-0.572, respectively).

**Conclusions:** These findings suggest that abdominal obese children have different plasma FFAs concentrations than non-obese peers, and the associations of FFAs with cardiometabolic risk factors are present in childhood.

**Keywords:** Students, body mass index, cardiometabolic risk factors, palmitoleic acid, dihomo-gamma linoleic acid.

(1)Aristizabal, Juan; (1)González-Zapata, Laura Inés; (2)Estrada-Restrepo, Alejandro; (3)Monsalve, Julia; (4)Restrepo-Mesa, Sandra; (5)Gaitan-Charry, Diego.

(1)PhD. Nutritionist. School of Nutrition and Dietetics, University of Antioquia, Medellín. Colombia; (2)MSc. Epidemiology. Grupo de Investigación en Demografía y Salud. Universidad de Antioquia. Colombia; (3)MSc. Nutritionist. Escuela de Nutrición y Dietética. Universidad de Antioquia. Colombia; (4)Professor. Colombia; (5)PhD. Doctor. Grupo de Investigación Alimentación y Nutrición Humana. Universidad de Antioquia. Colombia.

**Background and objectives:** In adults with abdominal obesity (AO), increased concentration of free fatty acids (FFAs) is associated with cardiometabolic risk factors. However, this association remains controversial in children. This study aimed to compare plasma FFAs concentration in children with and without AO, and to examine FFAs associations with cardiometabolic risk factors.

**Methods:** Twenty-nine children with AO were matched, one by one, according to sex and age, with 29 non-obese peers. Abdominal obesity was classified using the ninety percentile from the IDEFICS-Study (Identification and prevention of Dietary- and lifestyle-induced health EFects In Children and infantS). Fasting plasma glucose, insulin, and lipids were determined by colorimetric and enzymatic methods. Plasma FFAs were analyzed by gas chromatography.

**Results:** Seventeen boys and twelve girls with similar average age (7.1 ± 2.6 vs 7.2 ± 2.7 y; p>0.05) were included in each group. Children with AO showed higher body mass index (BMI) (19.7 vs 15.4 kg/m2; p<0.001), waist circumference (64.9 vs 53.7 cm; p<0.001), systolic blood pressure (98.6 vs 93.5 mmHg; p<0.01)

(1)Cho, Younok; (2)An, Jeehyun.

(1)PhD. Professor. Department of Food and Nutrition. Duksung women's university. Republic of Korea; (2)Department of Food and Nutrition. Duksung Women's University. Republic of Korea.

**Background and objectives:** Replenishment of body's fuel store after exercise is a key process for the next bout of exercise. This study was conducted to examine the effect of exercise-to-rest period ratio and physical training on the recuperation of stored fuel.

**Methods:** Forty rats weighing 95-105 g were randomly assigned to either non-exercise training (NT) or regular exercise training (T) groups for 5 weeks and were subdivided into two groups: non-exercise(NE), exercise and recuperation group(ER). NE group were sacrificed without exercise and ER group were sacrificed after 1 hour rest from exercising for 0.5(ER2:1), 1(ER1:1), 2(ER1:2) hours.

**Results:** Liver glycogen level was increased in ER group and was reached to the highest level at ER1:1 ratio while it was not different between NE group and ER group in NT animal. Muscle glycogen level was increased at ER2:1 ratio and returned to the level of NE in NT animal while it was reached to the highest level at ER2:1 ratio and decreased but still higher than those of the level of NE in T animal. Free fatty acid level was reached highest level at ER2:1 ratio and decreased and was lower than those of NE at ER1:1 and ER1:2 ratio in NT animal while it was reached to the highest level at ER2:1 ratio and decreased but was not lower than those of NE at ER1:2 ratio in T animal. Muscle triglyceride level of ER group was lower than that of NE group regardless of the different exercise-to-rest period ratio in NT animal while it was decreased and lower than that of NE group at ER2:1 and ER1:1 ratio but returned to that of NE in T animal. Muscle protein level was decreased at ER2:1 and started to be increased but was not reached to the level of NE group in NT animal while it was decreased.
creased at ER2:1 and ER1:1 but was returned to the level of NE group in T animal.

**Conclusions:** Animal would have to rest for periods in the 1:2 ratio in non trained group and 1:1 ratio in trained group to replenish the body's fuel store after exercise.

**Keywords:** exercise, rest, recuperation, training, fuel

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EFFECT OF EXERCISE-TO-REST PERIOD RATIO AND PHYSICAL TRAINING ON THE RECUPERATION OF ANTIOXIDATIVE DEFENSE SYSTEM IN RATS

(1)Cho, Younok; (2)Kang, Keumjee.

(1)PhD. Professor. Department of Food and Nutrition, Duksung women's university. Republic of Korea; (2)PhD. Professor. Department of Food and Nutrition. Duksung Women's University. Republic of Korea.

**Background and objectives:** Successful recuperation after exercise is a key process to prepare the athlete for the next bout of exercise and the major components of recuperation may be the recuperation of antioxidative defense system of the body. This study was conducted to examine the effect of exercise-to-rest period ratio and physical training on the recuperation of antioxidative defense system of tissue from exercise in animal.

**Methods:** Forty 4-week old male Sprague-Dawley rats weighing 95-105 g were randomly assigned to one of two groups: non-exercise training (NT) or regular exercise training (T). At the end of week 5, the rats were subdivided into two groups: non-exercise(NE), exercise and recuperation group(ER). NE group were sacrificed without having performed exercise at the end of week 5. ER group were sacrificed after 1 hour rest from exercising on a treadmill (15° incline, 0.5-0.8 km/hr) for 0.5(ER2:1), 1(ER1:1), 2(ER1:2) hours respectively. Catalase activity was determined in plasma. The activity of superoxide dismutase (SOD), ratio of reduced glutathione to oxidized glutathione (GSH/GSSG), and levels of malondialdehyde (MDA) were determined in liver cytosol.

**Results:** SOD activities of NT animal were decreased at ER1:2, ER1:1, ER2:1 ratio and was not recovered to that activity of NE group while that of T animal were decreased at ER2:1, ER1:1 ratio and was recovered to that activity of NE group at ER1:2 ratio. Catalase activities of both NT and T animals were not different at ER2:1, ER1:1, ER2:1 ratio from that of NE group. MDA levels of both NT and T animals were increased at ER2:1, ER1:1 ratio but decreased at ER1:2 ratio and were not recovered to level of NE group. The results of this study suggest that training may improve exercise performance by enhancing the antioxidative defense system.

**Conclusions:** Animal would have to rest for periods in the 1:2 in non trained group and 1:1 in trained group to recuperate an antioxidative defense system and this system is effective in exercise trained animals.

**Keywords:** exercise, rest, recuperation, training, antioxidative system

144/835

VALIDITY OF NUTRIENT INTAKES DERIVED FROM WEBSITE DISH-BASED DIETARY ASSESSMENT

(1)Matsuzaki, Eri; (2)Michie, Mikiko; (3)Kawabata, Terue.

(1)Master of Nutrition. Kagawa Nutrition University. Japan.;

(2)Nutritionist. Greenhouse Co., Ltd. Japan.; (3)Dr. Nutrition. Kagawa Nutrition University. Japan.

**Background and objectives:** We compared the relative validity of nutrient intakes derived from Internet website dish-based dietary records (WDDR) and weighed dietary records (DR).

**Methods:** The study participants were 218 registered users of a dietary management website and all were female. After excluding participants with missing data, 137 participants were included in the final analyses. The participants recorded their daily meals on both the DR and WDDR. The DR calculated energy and nutrient intakes using Standard Tables of Food Composition in Japan. For the WDDR, energy and nutrient intakes were automatically calculated by the website. We compared differences in the DR and WDDR with respect to intakes of energy and 13 nutrients.

**Results:** The median values for the intake of protein and seven nutrients showed no significant difference between the DR and the WDDR, and the estimated values were generally similar. The interquartile range of nutrients with a significant difference did not show a large deviation with the exception of vitamin A. Correlation coefficients showed a strong correlation of 0.7 with the exception of sodium and vitamin E. The kappa coefficients for energy, calcium, iron, vitamin C, and dietary fiber were good to fair and all others were moderate. Using Bland-Altman plots, we found proportional errors in intake values for vitamins and minerals. We could not confirm obvious systematic errors for energy, protein, fat, and carbohydrate intake.

**Conclusions:** Overall, the WDDR is considered to be a valid method of estimating energy and nutrient intake.

**Keywords:** web, dish-based, dietary records, Internet

**Conflict of Interest Disclosure:** Eri Matsuzaki, Mikiko Michie, and Terue Kawabata declare that they have no conflicts of interest. The authors whose names are listed certify that they have NO affiliation with any organizations or any entities in relation to financial interest.
# FERROGENIC ANEMIA: OXIDATIVE STRESS AND CYTOMOLECULAR DAMAGE ASSOCIATED WITH FERROUS SULFATE PREVENTIVE ADMINISTRATION

(1)Gambaro, Rocio Celeste; (2)Seoane, Analía; (3)Padula, Gisel.

(1)BSc. Anthropologist. Instituto de Genética Veterinaria "Ing. Fernando N. Dulout". Universidad Nacional de La Plata. Consejo Nacional de Investigaciones Científicas y técnicas. Argentina;

(2)PhD. Biologist. Instituto de Genética Veterinaria "Ing. Fernando N. Dulout". Universidad Nacional de La Plata. Consejo Nacional de Investigaciones Científicas y Técnicas. La Plata. Argentina;

(3)PhD. Anthropologist. Instituto de Genética Veterinaria "Ing. Fernando N. Dulout". Universidad Nacional de La Plata. Consejo Nacional de Investigaciones Científicas y Técnicas. Facultad de Ciencias Naturales y Museo UNLP. La Plata. Argentina.

**Background and objectives:** Iron deficiency anemia (IDF) is a pathological condition characterized by insufficient iron to synthesize iron-containing proteins and enzymes. Iron is an essential trace mineral for a number of biological functions. Although iron deficiency may be prevented by adequate dietary intakes, it is internationally agreed that iron supplementation to lactating mothers and infants is a priority. The Argentine Pediatric Society recommends daily ferrous sulfate supplementation as a preventive strategy. Since the 90's, a single weekly dose of ferrous sulfate has been an alternative treatment. On the other hand, ferrous sulfate is known for its pro-oxidant properties, leading to increased oxidative stress that may seriously damage membranes, proteins and DNA. The aim of this study was to determine the possible cytomolecular damage and oxidative stress caused by daily and weekly administration of ferrous sulfate for IDF prevention, in in vitro cultured human lymphocytes.

**Methods:** We performed two controls (negative and positive) and two ferrous sulfate treatments (weekly and daily). Cytomolecular damage was assessed by the comet assay and oxidative stress using T-BARS.

**Results:** Weekly ferrous sulfate treatment elicited a decreasing trend in the production of oxygen free radicals and, consequently, of oxidative stress and cytomolecular damage.

**Conclusions:** Since weekly ferrous sulfate supplementation causes less side effects, it would allow higher treatment compliance, thus decreasing IDF prevalence. Future in vivo studies and the development of further techniques would enable us to obtain more conclusive results.

**Keywords:** Iron deficiency anemia, T-BARS, Comet assay, Preventive treatment

# INVITRO AND IN VIVO EVALUATION OF THE FERMENTABILITY OF TWO DIFFERENT WHEAT-DE-RIVED FIBRE FRACTIONS

(1)Deroover, Lise; (2)Boets, Eef; (3)Gomand, Sara; (4)Preston, Thomas;

(5)Vermeulen, Karen; (2)De Preter, Vicky; (3)Courtin, Christophe;

(3)Delcour, Jan; (2)Verbeke, Kristin.

(1)MSc. Department of Clinical and Experimental Medicine. KU Leuven. Belgium.; (2)PhD. Department of Clinical and Experimental Medicine. KU Leuven. Belgium.; (3)PhD. Department of Microbial and Molecular Systems. KU Leuven. Belgium.; (4)PhD. Scottish Universities Environmental Research Centre. University of Glasgow. UK.; (5)MSc. Department of Pathology, Bacteriology and Avian Diseases. Ghent University. Belgium.

**Background and objectives:** Dietary fibre (DF) is neither digested nor absorbed in the small intestine but is fermented in the large intestine by the resident microbiota with production of short-chain fatty acids (SCFA). The extent of DF fermentation depends on structural and physicochemical characteristics of the fibre.

In this study, the SCFA production from two structurally different wheat-derived fibre fractions was quantified using in vitro fermentation experiments and a dietary intervention study in healthy humans.

**Methods:** Readily fermentable arabinoxylan oligosaccharides (AXOS) and water-insoluble dietary fibre-enriched bran (DF-B) were produced from both unlabelled and 13C-enriched wheat bran (WB). Both (unlabelled) fractions were incubated in vitro with faecal slurries obtained from healthy subjects at 37°C for 24h. Remaining arabinoxylan (AX) was quantified using GC-FID and metabolite profiles, including SCFA, were analysed using GC-MS. In the human intervention study, ten healthy subjects consumed 13C-labeled AXOS and 13C-labelled DF-B at breakfast on two separate days followed by collection of breath and blood samples on regular time points for 14h. Breath samples were analysed for 13CO2 and H2 and blood samples were analysed for 13C-SCFA levels. A faecal sample was analysed for butyrate-producing capacity.

**Results:** After in vitro incubation of faecal slurries with AXOS for 24h, only 5% of residual AX was retrieved whereas almost 90% of the AX remained after DF-B incubation. Fermentation of AXOS resulted in a faster and higher increase in SCFA compared to DF-B. Metabolite profiles showed that AXOS fermentation was associated with SCFA, some alcohols and aldehydes, whereas DF-B fermentation was associated with branched-chain and median-chain faty acids. In humans, excretion in breath of the fermentation gases, 13CO2 and H2 increased after 13C-AXOS but not after 13C-DF-B. Cumulative total 13C-SCFA levels in blood were significantly higher after 13C AXOS compared to 13C-DF-B (89 [74-103] mmol vs. 9.3 [5.4-15.5] mmol; p=0.005). Total butyrate recovery was not correlated to the butyrate-producing capacity in the faecal samples.

**Conclusions:** In conclusion, the molecular structure and physicochemical characteristics of AX in WB, clearly determine the extent of fermentation. Furthermore, the use of stable isotope labelled substrates provides an efficient manner to establish differences in in vivo fermentation.
FROM STARCH STRUCTURE TO ITS IN VIVO METABOLIC FATE: ADVANCED IMAGERY TECHNIQUES TO EXPLAIN THE CHANGES IN STARCH STRUCTURE DURING DIFFERENT BISCUIT-MAKING PROCESSES

(1)Cesbron Lavau, Gautier; (2)Goux, Aurélie; (2)Meynier, Alexandra;

(2)Vinoy, Sophie.

(1)Msc. Nutrition Scientist. Mondelez International; (2)PhD.

Mondelēz International R&D. Nutrition Research Department.
France.

**Background and objectives:** Based on WHO recommendations, starch represents the largest component of our daily energy intake (40% to 50%). During food manufacturing, heat, moisture and pressure alter dramatically the digestibility of starch in processed foods (Bornet et al. 1993; Vinoy et al. 2016). The control of these major process parameters can lead to large differences of starch digestibility (Englyst et al. 2003).

The originality of this work was to combine complementary measures to characterize the extent of starch gelatinization, its structure through sophisticated imagery as well as its rate of digestion measured in vitro (Englyst method) and in vivo (Glycaemic Index (GI) determination).

**Methods:** Four different technologies were chosen for their different process parameters: rotary biscuits, extruded cereals, soft cakes and rusks. The first three were made in two versions, wholegrain by addition of wheat bran and plain using regular wheat flour. Samples were sent for starch digestion analysis in vitro and a GI test using all 9 products was launched. Conjunctively, the internal structure and aspect of the starch were determined by using x-ray diffraction (XRD), x-ray micro-tomography and field emission gun scanning electron microscopy (FEG-SEM).

**Results:** The study highlighted several differences between technologies. Only the rotary products, with and without bran maintained a high slowly digestible starch (SDS) level (24g/100g, and 28 g/100g respectively) and a low GI (47 ± 5, and 43 ± 3 respectively). Soft cakes with and without bran showed low SDS values (2g/100g for both product) and medium GI values (63 ± 6, and 66 ± 5 respectively). Rusk products displayed low SDS values (1g/100g) and a medium GI (61 ± 4). The extruded cereals, with and without bran had the lowest SDS values (0.1g/100g for both products) and higher GI values (66 ± 4 and 77 ± 4 respectively). The imaging technologies unveiled significant changes in starch structure and showed a higher level of preservation for the rotaries which corresponded to the in vitro and in vivo digestion results.

**Conclusions:** By controlling food process, slow digestibility of starch can be maintained because of its starch structure preservation.

**Keywords:** Carbohydrate metabolism, Digestion, Wholegrain, Glycaemic index, Food Process

THE PROTECTIVE ROLE OF MITOCHONDRIAL SERINE HYDROXYMETHYLTRANSFERASE AGAINST OXIDATIVE STRESS-INDUCED MITOCHONDRIA DYSFUNCTION AND DNA DAMAGE

(1)Yeh, I-Ling; (2)Shane, Barry; (3)Huang, Rwei-Fen S.

(1)Dietitian. Department of Dietetics. National Taiwan University Hospital. Taiwan; (2)Professor. Nutritional Sciences & Toxicology. Department of Nutritional Sciences & Toxicology. University of California, Berkeley. USA.; (3)Professor. Department of Nutritional Science. Fu Jen Catholic University. Taiwan.

**Background and objectives:** Two different isoform enzymes serine hydroxymethyltransferase (SHMT) involved in folate metabolism occurs mainly in the cytosol and mitochondria (mt). Numerous studies suggested that polymorphisms of cytosol SHMT (cSHMT) were associated with related to plasma homocysteine levels and increased risks of cancer development. The functional role of mitochondrial SHMT (mSHMT) remains obscure. By use of the Chinese hamster ovary (CHO) mutant cells, glyA, with defective mSHMT, we have previously shown that treatment of glyA with tert-Butylhydroperoxide (tBH), an exogenous oxidative stress, resulted in elevated superoxide levels and depleted mt folate levels. Aims of the study were to determine the role of mSHMT and its interaction with folate insufficiency in mt function upon oxidative stress.

**Methods:** Treated with 50µM tBH for 72hrs and pre-incubation with folate supplements in glyA.

**Results:** Treated with 50µM tBH for 72hrs in glyA was showed that mt biogenesis increased including mtDNA content and mt mass. Additional, mtDNA-encoded genes also increased including cytochrome c oxidase subunit (COX) II, III and ATP synthase Fo subunit 6. However, it also increases 8-hydroxy-20-deoxyguanosine (8-OHdG), a DNA damage indication in glyA treatment via tBH. Intriguingly, pre-incubation with folate supplements increase both cytosolic and mt folate and reduced the t-BH-induced ROS. The folate supplementation was attenuated confusing mt biogenesis and 8-OHdG by t-BH treated glyA.

**Conclusions:** Taken together, the data suggest that functional defects of mSHMT in the mt, as observed in glyA, has handicapped the biosynthesis of folate, and folate in the mt plays a critical role in the protective action of exogenous oxidative stress.

**Keywords:** Folate, mitochondria, serine hydroxymethyltransferase, mitochondrial biogenesis, DNA damage
# BIOMARKER OF LONG-CHAIN N-3 PUFA AND RISK OF BREAST CANCER: ACCUMULATIVE EVIDENCE FROM AN UPDATED META-ANALYSIS OF EPIDEMIOLOGICAL STUDIES

(1)Yang, Bo; (2)Ma, Ai Guo; (2)Li, Duo.

(1)PhD. Nutritionist. Research Institute of Nutrition and Health. Qingdao University; (2)PhD. Nutritionist. Research Institute of Nutrition and Health. Qingdao University. China.

**Background and objectives:** Long-chain (LC) n-3 polyunsaturated fatty acid (PUFA) in different biospecimens (serum/plasma/erythrocyte/adipose tissue) as biomarkers of fatty acid intake can provide objective measures that reflect both dietary consumption and relevant biological processes. Nevertheless, results of epidemiological studies on biomarker of LC n-3 PUFA in relation to risk of breast cancer (BC) are inconsistent. We therefore conducted a meta-analysis to assess the up-to-date pooling effects based on prospective cohort and case-control studies.

**Methods:** The compositions of LC n-3 PUFA in erythrocyte membrane (EM) and adipose tissue (AT) were compared between BC cases and noncases as a standardized mean difference (SMD). Risk ratios (RRs) or odds ratios (ORs) with 95% confidence interval (CI) from prospective and case-control studies were pooled using a random-effects model in the highest vs. lowest categorical analysis, respectively.

**Results:** A database was derived from 19 eligible studies (10 cohorts with circulating LC n-3 PUFA as biomarker, 6 case-control with AT and 3 with EM), comprising 133,943 individuals (4,386 cases and 129,557 noncases). BC cases have significantly lower LC n-3 PUFA compositions in EM than noncases (SMD = -0.27; 95% CI: -0.41, -0.13; I2 = 28.10%), whereas no significant difference was found in AT (SMD = -0.15; 95% CI: -0.06, 0.36; I2 = 81.50%). In pooled analysis of cohort studies (n=10), circulating LC n-3 PUFA was significantly associated with reduced risk of BC (RR = 0.88; 95% CI: 0.78-0.99; I2 = 0.00%), and the summary RR was 0.85 (95% CI: 0.72-1.00; I2 = 0.00%) for 20:5n-3, 0.98 (95% CI: 0.81-1.19; I2 = 0.00%) for 22:5n-3, and 0.86 (95% CI: 0.76-1.02; I2 = 10.00%) for 22:6n-3, respectively. In pooled analysis of case-control studies (n=9), LC n-3 PUFA composition in EM was inversely associated with risk of BC (OR = 0.36; 95% CI: 0.18-0.70; I2 = 86.40%), whereas no significant association was observed for AT (OR = 0.91; 95% CI: 0.64-1.28; I2 = 29.90%).

**Conclusions:** Circulating LC n-3 PUFA was inversely associated with risk of BC, especially 20:5n-3 and 22:6n-3. Such findings support that LC n-3 compositions in circulating blood may be an independent predictive factor for BC.

**Keywords:** Fatty acid, PUFA, Biomarker, Breast cancer, Meta-analysis

# TRANSCULTURAL ADAPTATION OF THE EATING MOTIVATION SURVEY (TEMS) TO BRAZILIAN PORTUGUESE

(1)Moraes, Jéssica; (2)Koritar, Priscila; (3)Alvarenga, Marle.

(1)BSc. Nutricionista Departamento de Nutrição. Faculdade de Saúde Pública. Universidade de São Paulo. Brazil.; (2)MSc. Departamento de Nutrição. Faculdade de Saúde Pública da Universidade de São Paulo. Brazil.; (3)PhD. Departamento de Nutrição. Faculdade de Saúde Pública da Universidade de São Paulo. Brazil.

**Background and objectives:** When we think about food that are many reasons why the people eat what they eat. People can eat motivated by factors like hunger, the appearance of the food, desire, taste, price, to control weigh or maybe handle some emotions, because it is fashionable or because of the nutritional content - and still by cultural or family traditional reasons. Understanding these motivations is crucial to development of nutritional recommendations able to modify inappropriate and dysfunctional eating patterns. This understanding can be evaluated by instruments - among the existing ones, The Eating Motivation Survey (TEMS) allows the identification of the motivations to eat and food choices. Therefore, the aim of this study is to present the TEMS transcultural adaptation process for applicability in Brazilian population studies.

**Methods:** The process involved the conceptual and item equivalence; semantic equivalence by 2 translators, 1 linguist, 22 experts (response frequency) and 23 bilinguals (comparing the results by paired t test, Pearson Correlation and Intraclass Correlation Coefficient); and the operational equivalence carried out with 32 adult individuals.

**Results:** All these equivalences showed satisfactory results to use the scale in Brazil, which allows the TEMS application to evaluate the food choices motivations in our context.

**Conclusions:** All these equivalences showed satisfactory results to use the scale in Brazil, which allows the TEMS application to evaluate the food choices motivations in our context.

**Keywords:** Eating behaviour, motivation, cross-cultural comparison, questionnaires
MULTIVARIATE NUTRITIONAL ASSESSMENT
THROUGHOUT SEXUAL MATURATION

(1)Cumpian-Silva, Jéssica; (2)Rinaldi, Ana Elisa Madalena; (3)Mazzeti, Camila; (4)Sá, Beatriz Medeiros; (4)Santos, Aline Micaele Reis; (4)Cruz, Danielle Rocha; (5)Conde, Wolney Lisboa.

(1)MD. Nutricionista. Laboratório de Avaliação Nutricional de Populações. Faculdade de Saúde Pública (FSP). Universidade de São Paulo (USP). Brasil; (2)PHD. Professor Doutor. Nutricionista. Faculdade de Medicina. Universidade Federal de Uberlândia. Brasil; (3)MD. Nutricionista. Departamento de Nutrição. Universidade de São Paulo. Brasil; (4)Estudante Graduação. Universidade de São Paulo. Brasil; (5)PHD. Professor Doutor. Nutricionista. Departamento de Nutrição. Universidade de São Paulo. Brasil.

**Background and objectives:** The form of analysis can lead to confusion in the influence of sexual maturation in the assessment of nutritional status.

**Methods:** Data came from 680 adolescents surveyed in city of Piracicaba (SP, Brazil), 1134 adolescents surveyed in city of Sao Paulo (SP, Brazil) and 3321 adolescents American research National Health and Nutrition Examination Survey (NHANES III). Multivariate patterns of nutritional status were estimated using Principal Components Analysis. We set eigenvalues to 0.7 and loadings to 0.2. Sexual maturation was self-evaluated according to five Tanner stages.

**Results:** The first 4 components are responsible for 63.5% of the variability of Piracicaba, 71.13% NHANES and the first three components in Sao Paulo account for 81.45%. The first component of Piracicaba and NHANES was characterized by anthropometric variables especially skinfolds (triceps, subscapularis, suprailiac and thigh) and waist circumference. The data of Sao Paulo held the second component was characterized by the same anthropometric variables. The second component was characterized by sexual maturation in Piracicaba and NHANES (pubic hair, breast and external genitalia) beyond the age, height, weight and phase angle. In São Paulo, the component that marks the sexual maturation was retained in the first, and presented the same characteristics of other banks with the same markings variables. The third was characterized by cholesterol, triglycerides and glucose in Piracicaba and NHANES. The third and final component in Sao Paulo was characterized by body composition variable (phase angle). The fourth component mixed load presented for biochemical variables. The adequacy of the sample in relation to the degree of partial correlation between variables was estimated by the Kaiser-Meyer-Olkin in (KMO), which was 0.8031 Piracicaba, 0.8712 NHANES and 0.7713 Sao Paulo.

**Conclusions:** Multivariate analyses of nutritional status reveals many possible profiles whose cannot be properly described using usually a single univariate indicator, such as body mass index. We have no conflict of interest to declare.

**Keywords:** Adolescent; Nutritional Assessment; Sexual Maturation; Body Composition; Biomarkers.

VEGETARIAN DIET, NUTRITIONAL STATUS AND
CIRCULATING BRAIN DERIVED NEUROTROPHIC
FACTOR LEVELS IN ADULTS

(1)Pimentel, Carolina; (2)Philippi, Sonia; (3)Simomura, Viviane;

(4)Teodorov, Elisabeth.

(1)Nutricionist. PhD. Professor of Health Science Institute Paulista University. UNIP. São Paulo. Brazil.; (2)Nutricionist. PhD. Professor of Department of Nutrition. Public Health School. University of Sao Paulo. São Paulo. Brazil.; (3)Nutricionist. MSc. Paulista University. UNIP. São Paulo. Brazil.; (4)Biologist. Professor of Mathematics, Computing and Cognition Center. Federal University of ABC. Santo André. Brazil.

**Background and objectives:** Introduction: Recent studies have demonstrated that the Brain Derived Neurotrophic Factor (BDNF) is related to a possible endocrine role and also to nutritional factors, since they are involved in the regulation of energy metabolism and a positive correlation with the Body Mass Index (BMI). Objective: To evaluate nutritional status (waist circumference [WC] and BMI) and circulating BDNF levels in adult vegetarian individuals compared to omnivores.

**Methods:** A cross-sectional study was carried out with 96 subjects (56 vegetarians and 40 omnivores) adults. BMI was obtained by dividing body weight (kg) by squared height (m2). To obtain weight, we used an electronic scale (Welmy®, model R-110) and the stature was measured using a portable anthropometer (Estad® - Alturexata, 0,35m to 2,13m). The waist circumference (WC) was used to evaluate the body composition with the umbilical scar measurement using an inextensible metric tape with a Sanny® brand lock. The classification of the nutritional status according to BMI followed the cut-off points recommended by the WHO (1995) and classified as normal (18,5 kg/m2 to 24,9 kg/m2) and altered (> 24,9 kg/m2). The WC was classified according to the risk of metabolic complications associated with obesity, according to gender, considered normal for women, when <80 cm and altered or at risk if > 80 cm, for men normal WC was considered < 94 cm and altered or at risk >94 cm (WHO, 1998). BDNF levels were evaluated with the BDNF Emax ImmunoAssay System (Promega, Madison, WI, USA) according to the manufacturer's specifications and was classified according to the median (626.0 pg/mL).

**Results:** In relation to nutritional status, the VEGs are leaner (63,9 ± 10,4 kg vs 69,4 ± 14,6 kg, p=0,032), they have a lower BMI (22,5 ± 2,6 kg/m2 vs 25,0 ± 3,9 kg/m2, p< 0,001) and lower values of WC (81,8 ± 8,2cm vs 87,8 ± 10,9cm, p=0,003). Regarding the BDNF variable, there was no difference between vegetarians and omnivores (662,8 ± 276,5pg/ml vs 698,1 ± 314,9 pg/ml, p=0,563).

**Conclusions:** The results suggest that the vegetarian diet has a beneficial role in relation to the nutritional status of these individuals.

**Keywords:** Nutritional status; Brain Derived Neurotrophic Factor; Vegetarian diet.
# THE TRUE EFFECT OF SEXUAL MATURATION ON THE ASSESSMENT OF THE NUTRITIONAL STATUS OF ADOLESCENTS

(1)Cumpian-Silva, Jéssica; (2)Mazzeti, Camila; (3)Rinaldi, Ana Elisa Madalena; (4)Sá, Beatriz Medeiros; (4)Santos, Aline Micaele Reis; (4)Cruz, Danielle Rocha; (5)Conde, Wolney Lisboa.

(1)MD. Nutricionista. Laboratório de Avaliação Nutricional de Populações. Faculdade de Saúde Pública (FSP). Universidade de São Paulo (USP). Brasil; (2)MD. Nutricionista. Faculdade de Saúde Pública. Universidade de São Paulo. Brasil; (3)PHD. Nutricionista. Professor Doutor. Faculdade de Medicina. Universidade de Uberlândia. Brasil.; (4)Estudante Graduação. Universidade de São Paulo. Brasil; (5)PHD. Professor Doutor. Nutricionista. Departamento de Nutrição. Universidade de São Paulo. Brasil.

**Background and objectives:** The index used in analysis can lead to confusion about influence of sexual maturation in the assessment of nutritional status. We will estimate multivariate indices from a pool of body measures to describe nutritional status.

**Methods:** Data came from 833 adolescents surveyed in Piracicaba(Brazil) and 3321 in United States(NHANESIII). Principal Components Analysis was applied to define multivariate patterns of nutritional status from anthropometric measurements (weight, height, skinfolds, and waist circumference), body composition (phase angle) and biochemical exams (glucose, cholesterol, triglycerides and hemoglobin). To define multivariate patterns of nutritional status we set eigenvalues to 0.7 and eigenvectors to ±0.2. Each pattern was set as outcome in a mixed multilevel regression with two levels. In the individual level we included sex and in the second we included Tanner stages, age, sex and socioeconomic score.

**Results:** The first four components are responsible for 63.5% of the variability in Piracicaba and 71.1% in NHANESIII. All patterns were the same for both surveys. The first component was characterized by anthropometric variables; especially skin folds (triceps, subscapular, suprailiac and thigh) and waist circumference a pattern of adiposity. The second was characterized by sexual maturation (pubic hair, breast and gonad), age, height, weight and phase angle a pattern of linear growth process. The third was characterized by cholesterol, triglycerides and glucose a pattern of metabolic markers linked to adiposity. The fourth component was characterized by phase angle and hemoglobin (positive eigenvector) and glucose (negative eigenvector) patterns of metabolic markers linked to lean mass. The adequacy of the sample in relation to the degree of partial correlation between variables was estimated by Kaiser-Meyer-Olkin(KMO), which was 0,80. Piracicaba and 0,87 in NHANESIII. In mixed effects regression models, variance attributable to sexual maturation at level 2 was low; the individual fixed effects explained changes in body composition mostly. The two surveys Tanner's stages explained less than 20% of patterns that are unrelated to sexual maturation and explained more than 80% pattern two that is related to linear growth and sexual maturation.

**Conclusions:** The nutritional status in adolescents does not require adjustment by sexual maturation. Adolescent weight gain may be the source of confusion at the time of diagnosis.

**Keywords:** Adolescent; Nutritional Assessment; Sexual Maturation; Body Composition; Biomarkers.

# GLYCEMIC PROFILE AND CIRCULATING BRAIN DERIVED NEUROTROPHIC FACTOR LEVELS IN ADULT VEGETARIANS

(1)Pimentel, Carolina; (2)Philippi, Sonia; (3)Simomura, Viviane; (4)Teodorov, Elisabeth.

(1)Nutricionist. PhD. Professor of Health Science Institute Paulista University. UNIP. São Paulo. Brazil.; (2)Nutricionist. PhD. Professor of Department of Nutrition. Public Health School. University of Sao Paulo. São Paulo. Brazil.; (3)Nutricionist. MSc. Paulista University. UNIP. São Paulo. Brazil.; (4)Biologist. Professor of Mathematics, Computing and Cognition Center. Federal University of ABC. Santo André. Brazil.

**Background and objectives:** Introduction: Data from the literature have repeatedly demonstrated the beneficial role of the vegetarian diet in reducing risk factors for cardiovascular diseases, mainly because they positively modulate biochemical parameters, particularly those related to glycemic control and lipemia. Recently, animal studies have demonstrated the relationship of the Brain Derived Neurotrophic Factor (BDNF) and its relationship with increased insulin sensitivity and consequent lowering of blood glucose levels. Objective: To evaluate the glycemic profile (fasting glycemia) and circulating levels of BDNF in adult vegetarian individuals compared to omnivores.

**Methods:** A cross-sectional study was carried out with 96 subjects (56 vegetarians and 40 omnivores) adults. The biochemical parameters were obtained by means of blood collection, glycemia by the Automated Enzymatic Colorimetric method and the BDNF levels by BDNF Emax® ImmunoAssay System (Promega®, Madison, WI, USA), according to the manufacturer's specifications. For analysis, biochemical variables were classified as normal and altered using the following reference ranges for altered fasting glycemia ( > 110mg/dL) and for BDNF (median [626.0 pg/mL]). The analyzes were performed in Statistical Package for Social Sciences (SPSS) version 20.0 and for all a level of significance of 5% was considered.

**Results:** Vegetarians had lower fasting blood glucose levels when compared to omnivores (82,13 ± 9,11 mg/dL vs 85,8 ± 10,87 mg/dL, p=0,004). Regarding the BDNF variable, there was no difference between vegetarians and omnivores (662,8 ± 276,5 pg/ml vs 698,1 ± 314,9 pg/ml, p=0,563).

**Conclusions:** The results indicate that the vegetarian diet has a beneficial role in relation to the glycemic control of these individuals, research with a greater number of subjects is necessary to understand its relationship with BDNF levels.

**Keywords:** Brain Derived Neurotrophic Factor; Glycemia; Vegetarian diet.
# METIGENTITY, A NUTRIGENETIC APPROACH GUIDING INDIVIDUAL NEEDS AND PREFERENCES TOWARDS OPTIMAL AND PERSONALIZED NUTRITION

**Background and objectives:** Nutritional assessment is based on reliable information that combines nutrient exposure, its bioavailability and metabolic function. However, individual's response to dietary assessment can be highly variable, which has been associated with small genetic differences found throughout people's DNA, being SNP (single nucleotide polymorphisms) the most common. Therefore, implementation of personalized nutritional guidance requires the development of evidence-based biomarkers able to dissect the interaction between gene variants, nutrients and life-style aspects.

The objective has been the design and implementation of a nutrigenetic approach characterizing a set of gene variants focused on genetic susceptibility to obesity (Metigentity) and extended to analysis of sports performance (MetigentitySport).

**Methods:** A panel of genes were selected from studies showing association with obesity, sports performance or related issues (diabetes, hypertension, sliming pattern, endurance, motivation, etc). In addition, a selection of genetic variants playing a role in individual responses to nutrients or ergogenic aids was also included.

**Results:** Around 50 gene variants have been prioritized and incorporated at Metigentity. Identification of the number of alleles associated with the risk-benefit variants allowed the setting of specific genetic scores (GS) and assessment of people's genotype. Metigentity estimates GS for (1) risk to develop metabolic alterations associated with obesity (type 2 diabetes, hypertension, hypertriglyceridemia,...); (2) macronutrient metabolism performance; (3) eating behaviour; (4) gustative perception and nutrient/food preferences; (5) the impact of dietary components on biomarkers related to metabolic diseases; and (6) the ability to lose weight and to experience the rebound effect in response to dietary interventions. MetigentitySport analyzes around 30 gene variants and estimates (1) genetic capacity to practice aerobic endurance sports and power, sprint and speed sports; (2) psychological aptitude towards sports; (3) genetic susceptibility to suffer from sport injuries; (4) response to nutrients and supplements related to performance; and (5) the response to particular exercises in terms of performance and health. Holistic integration of all GS is elaborated and guidelines best suited to the individual's genotype is supplied.

**Conclusions:** The nutrigenetic approach Metigentity (http://www.metigentity.com/), and its extension MetigentitySport, constitute evidence-based biomarkers and are valuable tools for nutritionists guiding individual needs and preferences towards optimal and personalized nutrition.

**Keywords:** nutrigenetics, SNP, obesity, sports performance, personalized nutrition

**Further collaborators:** JM was the recipient of a Torres Quevedo contract co-financed by the Ministerio de Economía y Competitividad of Spain Government and the European Social Fund.

# THE INFLUENCE OF THE PEAKING TRAINING ON SALIVARY IGA SECRETION IN COLLEGE TRIATHLETE

**Background and objectives:** Some triathletes often feel sick in conditioning period (peaking period) toward the race. The aim of this study is to consider the method of conditioning athletes by investigating the changes of the immunity through examining the variation of salivary IgA secretion.

**Methods:** For the college students' triathletes of 33 males and 7 females who participate the race, the amount of salivary IgA about one month before the race were continuously measured. In addition, meal content and the amount of water intake and physical activity in the training were recorded and the relationship with salivary IgA secretion was evaluated.

**Results:** No significant change in nutrition intake during the test period was observed. Significant high level of water intake was shown only on the race day. The average amount of physical activity a week of two to three weeks before the race was the highest as 744.7 ± 51.5 kcal/day, and that of one to two weeks before the race was 513.2 ± 28.5 kcal/day. That of last week before the race was 305.5 ± 29.9 kcal/day, and this value was significantly lower than the initial value (P <0.05). In the subjects whose average amount of physical activity of the three days in one to two weeks before the race (pre-race 12 to 14 days) was more than 1000 kcal/day, the amount of salivary IgA secretion significantly reduced at one week before the race (P <0.05), and at the day before the race further reduction was observed (P <0.1). When the daily high-intensity exercises above a certain level two weeks before the race were performed, the salivary IgA secretion from the week to the day before the race reduced. Therefore, peaking method performed during a certain period before the race which contains high-intensity exercise weakens the mucosal immune function and induces the state which is easy to break a condition.

**Conclusions:** In the college students' triathletes, the amount of the salivary IgA secretion was decreased by the peaking carried

<footnote>(¹)Palou, Andreu; (²)Mercader, Josep; (³)Gil, Raul; (⁴)Serra, Francisca;</footnote>

<footnote>(⁵)Sabater Bibiloni, Agustín.</footnote>

<footnote>(¹)MBA. Executive Director. Alimentómica S.L. Spin off of the University of the Balearic Islands. Spain; (²)PhD. R+D Department. Alimentómica S.L. Spin off of the University of the Balearic Islands. Spain; (³)MsC. Consultant. Alimentómica S.L. Spin off of the University of the Balearic Islands. Spain; (⁴)Prof. PhD. Laboratory of Molecular Biology Nutrition and Biotechnology. Nutrogenomics. University of the Balearic Islands. Spain. CIBER Physiopathology of Obesity and Nutrition CIBEROBN. Spain; (⁵)University of the Balearic Islands. Spain.</footnote>

<footnote>(¹)Matsumoto, Megumi; (²)Satoh, Kentaro; (³)Kushi, Hidehiko;</footnote>

<footnote>(⁴)Hamuro, Koji; (⁴)Sakurai, Masao; (⁴)Saito, Hiroshi; (⁴)Tanaka, Ryohei;</footnote>

<footnote>(⁴)Saito, Takao; (⁴)Kohda, Noriyuki; (⁴)Hamada, Koichiro.</footnote>

<footnote>(¹)PhD. Nutritionist.; (²)MSc; (³)PhD., MD; (⁴)PhD.</footnote>
out toward the race. This was considered to be one of the causes
breaking the condition just before the race.

**Keywords:** IgA, triathlete, peaking training, conditioning

144/1154

EFFECT OF JAPANESE SOUP STOCK “DASHI” IN AUTONOMIC NERVOUS SYSTEM ACTIVITY AND MENTAL FATIGUE

(1) Yamazaki, Hanae; (2) Moritaki, Nozomi; (3) Inoue, Kazuo.

(1)Ph.D. Associate professor. Department of Food Science and Human Nutrition. Faculty of Agriculture. Ryukoku University Associate professor. Department of Food Science and Human Nutrition, Faculty of Agriculture. Ryukoku University. Japan;

(2)Grad Student. Laboratory of Nutrition Chemistry. Division of Food Science and Biotechnology. Graduate School of Agriculture. Kyoto University. Japan.; (3)Ph.D. Associate professor. Laboratory of Nutrition Chemistry. Division of Food Science and Biotechnology. Graduate School of Agriculture. Kyoto University. Japan.

**Background and objectives:** The Japanese soup stock, called “Dashi”, is used in many traditional Japanese cuisine and it’s taste and flavor assume a fundamental part in most of the dishes. Recently, some studies have indicated that a long-period intake of dashi brings beneficial influence on health. In this study, we focused on the instant effect of dashi intake on the autonomic nervous system activity and emotion related to a feeling of fatigue.

**Methods:** Healthy and non-smoking volunteers aged 20-25 were participated in this study. The subjects consumed assigned breakfast at 9:00, and the experiments were conducted between 10:30 and 11:30. Heart rate variability (HRV) was employed to assess the autonomic nervous system (ANS) activity. To measure degree of mental fatigue condition, the flicker test was used with 30 minutes simple calculation tasks “Uchida Kraepelin Test”. The subjective assessment for emotional states were also determined by a Visual Analog Scale (VAS) method.

**Results:** We observed that the increase of parasympathetic nerve activity at 15 minutes time point after intake of dashi, which means that the emotional condition supposed to shift to the phase of relaxing. Interestingly, only the inhalation of dashi odor also induced similar effect on ANS. The results of flicker test and VAS showed that dashi intake decreased an accumulation of a feeling of fatigue.

**Conclusions:** These results suggested that dashi has acute effects on improving the emotional states, and the distinctive odor of dashi plays an important role in these effects.

**Keywords:** dashi, autonomic nervous system activity, flicker test, visual analog scale, anti-fatigue

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AN INCREASE IN 4E-BP1 LEVEL MEDIATES TO ENHANCE TRIGLYCERIDE ACCUMULATION IN RAT LIVER UNDER PROTEIN MALNUTRITION

(1) Toyoshima, Yuka; (2) Tokita, Reiko; (1) Taguchi, Yusuke; (3) Yoshizawa, Fumiaki; (4) Takahashi, Shin-Ichiro; (5) Kato, Hisanori; (1) Minami, Shiro.

(1)PhD. Department of Bioregulation. Nippon Medical School. Japan.; (2) Bachelor. Department of Bioregulation. Nippon Medical School. Japan.; (3) PhD. Department of Agrobiology and Bioresources. Faculty of Agriculture. Utsunomiya University. Japan.; (4) PhD. Department of Animal Sciences. Graduate School of Agriculture and Life Science. The University of Tokyo. Japan.; (5) PhD. Corporate Sponsored Research Program “Food for Life”. Organization for Interdisciplinary Research Projects. The University of Tokyo. Japan.

**Background and objectives:** We have shown that feeding of a low-protein diet increases the amount of eukaryotic translation initiation factor (eIF) 4E-binding protein 1 (4E-BP1), a substrate of mammalian target of rapamycin complex 1 (mTORC1) in rat liver. There are increasing evidences that mTORC1 pathway controls lipid metabolism. However, the role of 4E-BP1 in the regulation of lipid metabolism is not clear. The present study was undertaken to examine whether 4E-BP1 is involved in the enhancement of hepatic triglyceride (TG) accumulation in rats fed a low-protein diet.

**Methods:** Experiment 1. In order to increase 4E-BP1 in the liver artificially, five-week-old rats were injected via their tail vein with adenovirus (Ad) expressing rat 4e-bp1 or LacZ and fed a standard chow. Seven days after Ad injection, their livers were collected.

Experiment 2. In order to knockdown hepatic 4E-BP1 under protein malnutrition, five-week-old rats were injected via their tail vein with Ad expressing rat 4e-bp1 short-hairpin RNA (sh4E-BP1) or non-specific short-hairpin RNA. Eight days after Ad injection, each Ad-injected group was divided into two groups. One was fed a 15% casein diet as a control diet and the other was fed a 5% casein diet (5C) as a low-protein diet for 4 days. On the 5th day, their livers were collected.

**Results:** Experiment 1. The 4E-BP1 overexpression did not change hepatic TG levels.

Experiment 2. The 5C-feeding increased 4E-BP1 levels in the liver of both Ad-injected rats, but 4E-BP1 knockdown by sh4E-BP1 Ad injection suppressed 5C-feeding-induced increases in hepatic 4E-BP1 levels. The 4E-BP1 knockdown also suppressed 5C-feeding-induced increases in the protein levels of eIF4G, a translation initiation scaffolding protein, in the liver. Hepatic TG levels were significantly increased by 5C-feeding in the liver of both Ad-injected rats. However, 4E-BP1 knockdown attenuated 5C-feeding-induced hepatic TG accumulation.

**Conclusions:** These data indicate that TG accumulation is not enhanced just by increasing 4E-BP1 in the liver, although the increase in 4E-BP1 levels is necessary to enhance TG accumulation in the liver under protein malnutrition. In addition, it is suggested that 4E-BP1 might control TG levels in concert with eIF4G in rat liver under protein malnutrition.

**Keywords:** protein malnutrition, liver, 4E-BP1, triglyceride, rat
RISK OF EATING DISORDERS IN PATIENTS WITH
CELIAC DISEASE

(1)Babio Sánchez, Nancy Elvira; (2)Alcázar, Mireia; (3)Castillejo,
Gemma; (2)Recasens, Miriam; (4)Martínez-Cerezo, Francesc; (5)Vaqué,
Cristina; (5)Vila-Martí, Anna; (5)Torres-Moreno, Miriam; (2)Barrubés,
Laura; (6)Salas-Salvadó, Jordi.

(1)PhD. BSc. Nutritionist. 1Human Nutrition Unit. Faculty of Medicine and Health Sciences. Biochemistry and Biotechnology Department. Rovira i Virgili University. Sant Joan Hospital de Reus. IISPV. Reus. 2. CIBEROBN. Instituto de Salud Carlos III. Spain; (2)RD. Human Nutrition Unit. Faculty of Medicine and Health Sciences. Biochemistry & Biotechnology Department. Rovira i Virgili University and Sant Joan de Reus Hospital. IISPV. Reus. Spain; (3)MD. Pediatrics Gastroenterology Unit. Sant Joan de Reus Hospital, Pediatrics Research Unit. Rovira i Virgili University. IISPV. Reus. Spain; (4)MD. Service of Adult Gastroenterology. Sant Joan de Reus Hospital. Reus. Spain; (5)PhD. BSc. Faculty of Health Sciences and Welfare. Research Group of Food, Health and Welfare. University of Vic-Central University of Catalonia. Spain; (6)MD. PhD. Human Nutrition Unit. Faculty of Medicine and Health Sciences. Biochemistry & Biotechnology Department. Rovira i Virgili University and Sant Joan de Reus Hospital. IISPV. Reus. CIBEROBN. ISCIII. Spain.

**Background and objectives:** Several cases of eating disorders (EDs) were reported in patients with celiac disease (CD), suggesting that ED could be a comorbidity associated with CD. However, few epidemiological studies have assessed this potential association. We aimed to evaluate the risk of EDs in individuals diagnosed with CD in comparison to healthy controls.

**Methods:** A total of 98 cases and 98 controls matched by gen-
der, age and body mass index between 10 and 23 years-old were
studied. A questionnaire was completed on medical history, and
sociodemographic and anthropometric characteristics. Various
ED screening self-reported tests were administered.

**Results:** A total of 61.2% of the study population were girls with a mean age of 15.3±3.7 years-old. Patients with CD scored non-significantly higher on all the ED screening tests than control participants. No differences were observed between study groups in terms of the frequency of individuals who exceeded the clinical cutoff identifying those at risk of ED. Being a patient with CD above 13 years-old was associated with a 2.15 point increase in the Eating Attitude Test (EAT) score compared to being a control [βcoefficient=2.15 SE 1.04; P=0.04] after adjusting for various confounders.

**Conclusions:** Although being a patient with CD was associ-
ated with a significantly higher EAT score in individuals above 13
years old, no clear differences were observed between individuals
with CD and controls in terms of risk of ED when other screening
tests were used. More studies with larger samples and prospective
designs are warranted to confirm these findings.

**Keywords:** Celiac disease, eating disorders, Eating Attitude Test.

**Further collaborators:**

Vanessa Gutiérrez-Pensadoa. RD; Enric Sánchez. RD

A RANDOMIZED TRIAL OF IRON-BIOFORTIFIED
BEANS IN SCHOOL CHILDREN IN MEXICO

(1)Finkelstein, Julia; (2)Mehta, Saur; (3)Luna, Sarah V.; (3)Rahn, Maike;

(4)Villalpando, Salvador; (5)Mundo Rosas, Veronica; (6)Shamah-Levy, Teresa; (7)Beebe, Stephen E.; (8)Haas, Jere D.

(1)ScD. Assistant Professor. Division of Nutritional Sciences. Cornell University. USA; (2)ScD. Associate Professor. Division of Nutritional Sciences. Cornell University. USA; (3)PhD. Division of Nutritional Sciences. Cornell University. USA; (4)MD PhD. Instituto Nacional de Salud Publica, Cuernavaca. Mexico; (5)MSc. Instituto Nacional de Salud Publica, Cuernavaca. Mexico; (6)PhD. Instituto Nacional de Salud Publica, Cuernavaca. Mexico; (7)PhD. Centro Internacional de Agricultura Tropical. Cali. Colombia; (8)PhD. Professor. Division of Nutritional Sciences. Cornell University. USA.

**Background and objectives:** The objective of this randomized efficacy trial was to determine the effects of iron-biofortified beans (Fe-Beans) on iron status in children, compared to control beans (Control-Beans).

**Methods:** A randomized trial of biofortified beans (Phaseolus vulgaris L), bred to enhance iron content, was conducted for six months in Oaxaca, Mexico. Participants were school-aged children (5-12 y) attending 20 rural, public, boarding schools from the Mexican state of Oaxaca. Iron status [hemoglobin, serum ferritin (SF), soluble transferrin receptor (sTfR), and total body iron (TBI)], inflammatory biomarkers (C-reactive protein (CRP) and -1 acid glycoprotein (AGP)), and anthropometric indices were evaluated at enrollment and at the end of the trial. The primary outcomes evaluated included: hemoglobin, serum ferritin, and sTfR concentrations; anemia (Hb <12.0 g/dL) and iron deficiency (serum ferritin <15.0 µg/L); and total body iron. Hemoglobin concentrations were adjusted for altitude, and serum ferritin levels were adjusted for inflammation (CRP, AGP) using Thurnham and BRINDA methods. Total body iron was calculated using Cook's equation. Mixed models were used to examine the effects of Fe-Beans on hematological outcomes, compared to Control-Beans, adjusting for baseline indicator, with school as a random effect.

**Results:** At baseline, 17.8% of children were anemic (Hb <12.0 g/dL) and 15.9% were iron deficient (serum ferritin <15.0 mg/L). A total of 14.5% of children had elevated CRP (>5.0 mg/L) or AGP (>1.0 g/L) concentrations at baseline. During the randomized efficacy trial, indicators of iron status, including hemoglobin, serum ferritin, sTfR, and TBI concentrations improved from baseline to endline (6 months) in both intervention and control groups. However, Fe-Beans did not significantly improve iron status indicators, including hemoglobin, serum ferritin, sTfR, or total body iron, compared to Control-Beans. Similarly, there were no significant effects of the iron-biofortified beans intervention on anemia or iron deficiency, compared to the control beans.

**Conclusions:** In this 6-month randomized efficacy trial of iron-biofortified beans in school children in Mexico, indicators of iron status improved in both intervention and control groups. However, there were no significant differences in the effects of Fe-Beans on iron status, compared to Control-Beans.

**Keywords:** Iron, biofortification, anemia, iron deficiency, Mexico
144/1202

MUSCLE STRENGTH, METABOLIC PROFILE, SARCOPENIA AND SARCOPENIC OBESITY IN OLDER CUBAN PEOPLE. PRELIMINARY RESULTS

**Background and objectives:** Background: Sarcopenia is the age-associated loss of skeletal muscle mass and function; and the sarcopenic obesity may be defined by a high fat to fat-free mass ratio. These conditions can be accompanied by reduced muscle strength and metabolic disorders due to a decline of both number and fibre size of the muscle. Objectives: To evaluate variations on the muscle strength and metabolic profile associated to sarcopenia and sarcopenic obesity in elderly people.

**Methods:** A sample of adults older than 60 years, not institutionalized or attending day care institutions for the elderly, was evaluated to get information on body composition, blood glucose and lipids, and muscle strength. Muscle strength was measured by dynamometry; body fat, by electrical bioimpedance; and skeletal muscle mass (SMM) and skeletal muscle index (SMI), by Baumgartner equations. Univariate and multivariate statistical analysis were used.

**Results:** The frequency of sarcopenia was similar between men and women. Older individuals were more likely to have this condition. Body Mass Index (BMI), SMM and SMI were lower in sarcopenic subjects. Grip strength was markedly decreased in sarcopenic men, while no significant difference was found within women. Body fat was higher in sarcopenic men and not in sarcopenic women. Therefore, Sarcopenic obesity was more frequent within men. Subjects with this condition in both sexes were older, had higher BMI and body fat; and lower SMM, SMI and grip strength. The metabolic profile showed no differences between subjects with or without sarcopenia and was not associated with sarcopenic obesity either.

**Conclusions:** Reduced muscle strength is evident in both sarcopenia and sarcopenic obesity, and was more marked as the age increases. The lack of association between sarcopenia, sarcopenic obesity and metabolic disruptions must be further analysed using multivariate analysis and include treatment for metabolic diseases, physical activity and diet.

**Keywords:** elderly, sarcopenia, sarcopenic obesity, metabolism, strength.

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FOOD OR WATER FOR SANITATION: EXPLAINING REGIONAL DIFFERENCES IN CHILD MALNUTRITION RATES IN UGANDA

**Background and objectives:** Child undernutrition rates remain high across Uganda. However, there are important regional differences in malnutrition rates, with the southwestern region—described as the country's food basket—persistently recording disproportionately higher levels of child undernutrition compared to, for example, the northern region which has higher poverty and food insecurity linked to many years of civil war. It is important for policymakers to understand context-specific determinants of poor nutrition and not assume that higher agricultural productivity alone can solve such problems.

This study analyzes factors explaining differential child stunting rates between Southwestern and Northern Uganda.

**Methods:** Using panel data collected from two rounds of household surveys in rural Uganda in 2012 and 2014, the analysis includes >3,300 households (in each round), containing >5,400 children below 5 years of age. In addition to child anthropometry, the data contains variables on agriculture, socioeconomic, as well as on water, sanitation and health practices. Oaxaca decomposition methods, controlling for time and locational fixed effects, were used to examine factors that drive mean differences in height-for-age z-scores (HAZ) between regions.

**Results:** Mean HAZ scores were -0.95±0.03 and -1.77±0.06 for northern and southwestern Uganda, respectively. This represents a significant mean difference of 0.82±0.07 (p<0.001). Decomposition analysis reveals major differences in water access/volumes and important divergence in levels of food output. Households in Northern Uganda on average use 8 more liters (p<0.001) of water per capita per day for domestic purposes than households in the southwest. This difference alone accounts for 0.40±0.07 (p<0.001) in variability in HAZ scores observed. Interacting domestic water use with sanitation patterns showed that northern households are likely using domestic water for sanitation related habits, such as washing hands after toilet use.

**Conclusions:** Contrary to conventional wisdom, this study showed that differentials in undernutrition across regions of Uganda are not necessarily due to differences in food production or diet diversity, but are heavily driven by differences in domestic water use. This does not negate the need to address serious problems of food insecurity across the country, but the findings suggest

<footnote>(1)Ruiz-Alvarez, Vladimir; (2)Díaz-Sánchez, María Elena; (3)Hernández-Triana, Manuel; (4)Llera-Abreu, Elisa; (5)Díaz-Fuentes, Yoandry;</footnote>

<footnote>(4)Rogue Treville, Celia María; (5)Lanyau Domínguez, Yeneisy;</footnote>

<footnote>(5)Campos-Hernández, Derbis; (4)Díaz-Domínguez, Maybe;</footnote>

<footnote>(5)Hernández-Hernández, Hector.</footnote>

<footnote>(1)Msc. Researcher; (2)PhD. Nutrition Center. National Institute of Hygiene, Epidemiology and Microbiology. Cuba.; (3)PhD. International Institute for Healthcare Professionals. USA.; (4)BSc. Nutrition Center. National Institute of Hygiene, Epidemiology and Microbiology. Cuba.</footnote>

<footnote>(1)Kabunga, Nassul; (2)Ghosh, Shibani; (3)Duggan, Christopher;</footnote>

<footnote>(4)Bashaasha, Bernard; (5)Griffiths, Jeffrey; (2)Webb, Patrick.</footnote>

<footnote>(1)PhD. Research Fellow. International Food Policy Research Institute. Kampala Office. Uganda.; (2)Feed the Future Innovation Lab for Nutrition. Friedman School of Nutrition Science and Policy. Tufts University. Boston. USA.; (3)Department of Global Health and Population. Harvard T.H. Chan School of Public Health. Harvard University. USA.; (4)College of Agriculture and Environmental Sciences. Makerere University. Kampala. Uganda.; (5)Department of Public Health and Community Medicine. School of Medicine. Tufts University. Boston. USA.</footnote>
a need for greater attention to WASH in development programming alongside agricultural productivity enhancement.

**Keywords:** Child malnutrition, height-for-age (HAZ), domestic water use, sanitation, Uganda.

(1)Mazzeti, Camila; (1)Cumpian-Silva, Jéssica; (2)Rinaldi, Ana Elisa Madalena; (3)G Carvalho, Julia; (4)Lisboa Conde, Wolney.

(1)MD. Nutricionist. School of Public Health. University of Sao Paulo. Brazil.; (2)PhD. Nutricionist. Federal University of Uberlândia.; (3)Student of Nutrition. School of Public Health. University of Sao Paulo. Brazil.; (4)PhD. Nutricionist. School of Public Health. University of Sao Paulo. Brazil.

**Background and objectives:** Allometry presents an alternative for to discuss and analyze the weight and height gains during the growth, monitoring the peculiarities of physical growth and changes in the body for a good nutritional assessment. To propose a new allometric nutritional index based on the weight-height relation and to evaluate its performance according to Benn's premises.

**Methods:** Individuals under 19 years old from Brazil, USA, South Korea, Mexico and England. The cleaning excluded values under or above 2SD of the residues of the regressions weight-Age, height-Age and weight-height by country, gender and age. The coefficient b0 of the log-linear weight-height regressions were used as exponent at each age for the calculation of the new international allometric body mass index (IMCA). The exponent was modeled between the ages by spline. Associations between traditional BMI (IMCT) or IMCA with height and adiposity were estimated by the Pearson coefficient.

**Results:** Cleaning excluded 8.5% individuals from the data pool. The value of b0 is 2 at birth in both genders and evolves to values of 3 at the beginning of puberty (in different moments for boys and grils), gradually regressing to the value 2 at the end of growth. The association between IMCA and height was zero and higher than 0.8 with total adiposity in all age groups. In the same comparison, the associations with IMCT were near to 0,4 in some genders and ages, and above 0.8 for association with adiposity.

**Conclusions:** The exclusion of extreme values of the sample provided b0 exponents more consistent than in other published works. The IMCA showed a superior performance to that recorded for the IMCT in the two premises of Benn and revealed an index not influenced by height, which makes it more suitable to analyze the nutritional status of children and adolescents under 19 years. The new IMCA requires additional studies to determine new cutoff points.

**Keywords:** Allometry; BMI; Growth; Nutritional Evaluation; Adiposity

(1)Pinto-e-Silva, Maria Elisabeth Machado; (2)Santos, Fernanda Ferreira; (3)Simoni, Natalia Koren; (4)Gines Geraldo, Ana Paula.

(1)Assoc. Professor. School of Public Health. University of São Paulo. São Paulo. Brazil; (2)BSc. Nutr Depart. School of Public Health. University of São Paulo. Brazil.; (3)MSc. Nutr.Depart. School of Public Health. University of São Paulo. Brazil.; (4)PhD. Nutr. Depart. Federal University of Santa Catarina. Brazil.

**Background and objectives:** BACKGROUND: Social medi- as has increasingly been used by individuals from Brazil's different regions and social strata, what turn them relevant to communicate nutritional knowledge in a planned and referenced way. Its use can be optimized by nutrition and marketing knowledge interdiscipli- narity.

**OBJECTIVES:** It aims to observe the effects of nutrition and marketing knowledge interdisciplinarity during planning and exe- cution of publications related to nutrition, available on Facebook's DIETÉCNICA page; and its metrics variations in two distinct mo- ments.

**Methods:** Metrics provided by Facebook's DIETÉCNICA page were compared between 2013 and 2015, in which publica- tions development and execution were not planned, and between 2015 and 2017, applying "Digital Marketing Decalogue". This tool consists of ten steps that are used in accordance with the time in which the media has been. In this project was used "social net- works", "search", "planning", "blogs", "final touch" and "results" for contents planning and execution, decision of publication frequen- cy and collection of its metrics. Metrics are composed by number of people who view the publication; likes, clicks, comments and shares.

**Results:** As a result of the use of "Digital Marketing Decalogue" there was a greater increase on DIETÉCNICA's metrics. Without the use of the marketing tool, number of views on publications (range) was around 300, after the its implementation it achieved about 3000, demonstrating the relevance of planning publications. It was observed that nutrition and marketing knowledge interdis- ciplinarity during publications planning and execution contrib- uted positively to average increase on Facebook metrics, when compared with the moment when there was no interdisciplinarity.

**Conclusions:** With this project is possible to observe that knowledge interdisciplinarity is essential for planning interven- tions of nutritional information communication on social media, such as Facebook, because it enables directing efforts to achieve a common goal; in this project it was a greater communication of nutritional knowledge for media users, which was achieved.

**Keywords:** Nutrition, Communication of Nutritional Infor- mation, Digital Marketing, Social Media.
## GENETIC POLYMORPHISMS IN THE AHR AND ADORA2A GENES ARE ASSOCIATED WITH COFFEE CONSUMPTION

(1)Miranda, Andreia; (2)Steluti, Josiane; (2)Carvalho, Aline M.; (2)Fisberg, Regina; (2)Marchioni, Dirce.

(1)PhD student. Nutritionist. Department of Nutrition. School of Public Health. University of São Paulo. SP. Brazil.; (2)PhD. Nutritionist. Department of Nutrition. School of Public Health. University of São Paulo. Brazil.

**Background and objectives:** Coffee is among the most widely consumed beverages in the world and has received considerable attention regarding health risks and benefits. Recent epidemiological studies identified single nucleotide polymorphisms (SNPs) in aryl-hydrocarbon receptor (AHR) and adenosine A2A receptor (ADORA2A) genes that are associated with caffeine and coffee consumption. The aim of this study was to examine whether the genetic polymorphisms from the AHR and ADORA2A gene regions were associated with usual coffee consumption in the Brazilian population.

**Methods:** Data came from the 'Health Survey of Sao Paulo' (ISA-Capital 2008). The study sample included 540 participants aged 20 years or older, in Sao Paulo City, Brazil. Diet was assessed by two 24-hour dietary recalls (24HR) and usual intakes were calculated using the Multiple Source Method. Coffee consumption was categorized into <1, 1-3, and ≥3 cups/day. DNA extraction occurred by salt method, with the support of a mass spectrometry-based detection for quantification. Three SNPs (rs2298383, rs4410790, rs6968865) using the CRP-based assays were genotyped. Multiple logistic regression models were performed to estimate the association between SNPs and categories of coffee consumption, adjusting for potential confounders i.e. age, sex, race, body mass index, smoking status, physical activity level, alcohol consumption, and total energy intake.

**Results:** The final study population showed a mean age of 44.9 (SE 1.1) years, mostly women (54.3%), self-declared white (60.1%), never or former smokers (76.8%), insufficiently active (77.4%), and with overweight/obesity (55.5%). The mean of coffee consumption was 141.1 mL/day. After multivariate adjustment, significant and positive associations were observed between coffee consumption (≥ 3 cups/day) and ADORA SNP (rs2298383) and AHR SNPs (rs4410790, rs6968865). Compared with subjects who consumed ≤1 cup coffee/day, subjects who consumed ≥3 cups/day were more likely to be carriers of mutant allele. The corresponding odds ratio (OR) and 95% confidence interval (CI) were 1.68 (1.01, 2.82), P-trend=0.047; 1.84 (1.07, 3.17), P-trend=0.025; and 1.71 (1.03, 2.84), P-trend=0.037 for rs2298383, rs4410790, and rs6968865, respectively.

**Conclusions:** Our findings show that genetic variations in the ADORA2A and AHR receptor gene polymorphisms were associated with coffee consumption (≥ 3 cups/day).

**Keywords:** coffee consumption; ADORA2A, adenosine A2A receptor gene; AHR, aryl-hydrocarbon receptor gene; polymorphisms; genotype.

## DIFFERENT ISOMERS OF TOCOPHEROL EVOKE DISSIMILAR CYTOSOLIC CALCIUM INCREASES AND ANTIOXIDANT IMPACT IN LIVING CACO-2 CELLS

(1)Hidalgo, Miltha; (2)Rodríguez, Vania; (3)Porras, Omar.

(1)Nutricionist. Instituto de Nutrición y Tecnología de los Alimentos. Universidad de Chile. Chile; (2)Physician. Instituto de Nutrición y Tecnología de los Alimentos de la Universidad de Chile. Chile; (3)PhD. Instituto de Nutrición y Tecnología de los Alimentos de la Universidad de Chile. Chile.

**Background and objectives:** Tocopherols are hydrophobic bioactive compounds, which isomers together are considered as vitamin E. Particularly abundant in seeds and nuts, each isomer is present at different proportion, being the α-tocopherol the most abundant. However, comparisons of biological impact of each individual isomer are scarce; therefore we have evaluated the acute impact of α, γ and δ-isomers on intestinal epithelial cell line, Caco-2.

**Methods:** Cell line Caco-2 was used in the passages 40-60. Living cell imaging was accomplished by epifluorescent microscopy technique in an open chamber modality. Cytosolic calcium recordings were possible by loading the cells with fura-2, a fluorescent calcium indicator. Antioxidant cellular capacity was achieved by transducing HyPer biosensor, a fluorescent protein able to sense intracellular H2O2. This study also determined fluctuations in intracellular sodium and pH by SBFI and BCECF, respectively.

**Results:** Acute exposure with tocopherol isomers evoked a rapid and consistent increase in cytosolic calcium by Caco-2 cells at concentration as low as 5 µM, with slight differences between them. Removal of extracellular Ca2+ was only effective to reduce the number of responsive cells by α-tocopherol but did not affect the calcium response evoked by γ and δ isomers. On the other hand, redox impact of tocopherol isomers was evaluated in Caco-2 cells expressing the HyPer biosensor, α- and δ- were the isomers that evoked bigger changes in the biosensor signal than γ-tocopherol. By mimicking a rapid cytosolic increase with ionomycin and monensin, both ionophores with different intrinsic mechanisms, we also could observe a deflection in HyPer signal, suggesting a role for cytosolic calcium in activate redox cellular machinery.

**Conclusions:** The different isomers of tocopherol elicited an acute redox response in Caco-2 cells, along with this cellular response we also detected a fast increase in cytosolic Ca2+, which had an extracellular origin only for α-tocopherol, whereas that calcium responses evoked by γ and δ isomers were not affected by extracellular Ca2+ removal. By inducing a massive entry of Ca2+ we were also able to record a redox response, suggesting that fluctuations in cytosolic Ca2+ are enough to mediate a redox response in living cells.

**Keywords:** tocopherol, Caco-2, Calcium imaging, HyPer, antioxidant capacity

**Further collaborators:** supported by CONICYT-FONDEF IT15I10048
ASSOCIATIONS BETWEEN FOOD INTAKE, PERCEIVED EXERTION, SUM OF 6 SKINFOLDS AND LEG CROSS-SECTIONAL AREAS IN PROFESIONAL ARGENTINEAN CYCLISTS ASSESSED BETWEEN MARCH-AUGUST 2016

Leonardi, Antonella.

PhD. Nutritionist. Department Nutrition Sciences. Maimónides University. Argentina.

**Background and objectives:** Road Cycling is known to be one of the most physically demanding sports, where athletes aim to achieve optimum muscle and fat parameters through a meticulous training and nutrition, which are crucial when seeking high performance. The aims of this study were to explore and relate Professional Cyclist's diet in terms of Carbohydrates, Proteins and fats, their perceived exertion during competition and their fat and muscle parameters.

**Methods:** Argentinean professional road cyclists were studied. 104 active elite road cyclists were assessed in terms of Age, Weight and Height, sum of 6 Skinfolds and leg cross-sectional areas, in order to quantify adipose and muscular tissues and compare with optimum standards. Perceived Exertion was inquired regarding a competition. Food Intake was evaluated through a 3-day register. Data was assembled in SARA System to assess nutrition in terms of calories, carbohydrates, proteins and fats. The rest of the variables were processed in order to create a full-descriptive statistical analysis and seek possible association between them through the x2 test.

**Results:** 40% of the cyclists reported a high sum of 6 skinfolds (55,2mm ±14,6mm). Diet was deficient in terms of energy and carbohydrates, and excessive in proteins. Significative association was found between an elevated sum of 6 skinfolds and a high perceived exertion (p<0,0001). An adequate consumption of energy (p=0,002) and carbohydrates (p=0,00001) were associated to a lower range of perceived exertion. An optimum sum of 6 skinfolds was associated to an adequate energy intake.

**Conclusions:** Nutrition assessment is needed to promote proper eating habits according to the demands of Argentinean professional cyclists, in order to reach optimum fat and muscle ratios and to a maximum performance. It is important that Argentinean cyclists begin to cover their high carbohydrate needs rather than exceeding their protein daily intake.

**Keywords:** Sport Nutrition, Cycling, Anthropometry, Perceived Exertion.

**Conflict of Interest Disclosure:** Identifying the feeding practices of Argentinean cyclists and their anthropometric profile, as well as the way these affect effort perception, could be useful not only approach a diagnosis, but also to estimate possible improvements in performance by developing more accurate diet patterns, looking for a morphological profile that adjusts better to professional road cycling.

INTERMITTENT FASTING IN DIFFERENT PERIODS OF PREGNANCY: MATERNAL EFFECT AND METABOLIC PROGRAMMING OF THE INSULINIC HYPOTHALAMIC VIA AND OBESITY IN THE MALE PROLE

(1)Geminiani A. Baptista, Julia; (2)Rosas Fernández, Mariana Alejandra; (3)Oliveira Batista, Leandro; (3)Tibau Albuquerque, Kelse.

(1)Nutritionist. Postgraduate Student. Rio de Janeiro Federal University. Macaé. Brazil.; (2)MSc. Nutritionist. Postgraduate Student. Rio de Janeiro Federal University. Macaé. Brazil.; (3)PhD. Nutritionist. Researcher in Experimental Nutrition Laboratories. Rio de Janeiro Federal University. Macaé. Brazil.

**Background and objectives:** Chronic dietary restriction during gestation may promote changes in fetal metabolism and impair the hypothalamic signaling of insulin in the offspring. Nonetheless, nausea and vomiting are common during gestation and induce intermittent feeding. However, it is not clear whether this intermittent involuntary fasting causes harm to offspring in adult life. We evaluated whether intermittence between feed restriction and ad libitum (IFR) in pregnant Wistar rats is capable of inducing obesogenic changes in male offspring.

**Methods:** Pregnant rats were kept with free access to chow (the control group, C) or with 60 % restriction of C on alternate days during the first half (fasting 1 group, F1) or in the second half (fasting 2 group, F2) or in whole pregnancy (total fasting group, TF). In lactation, the diet was ad libitum. After the period of lactation, the progenitors were euthanized for visceral fat collection, while the male offspring were maintained until adulthood (100 days) with ration ad libitum. Part of the adult offspring received intrahypothalamic cannula for administration of insulin (20 mU) or vehicle (0.9 % saline) and evaluated visceral adiposity.

**Results:** The offspring C responded to intracerebroventricular infusion of insulin, but F1, F2 and TR did not reduce food intake after 12 h and 24 h of insulin injection versus vehicle infusion. The maternal weight gain during pregnancy did not differ, but the cumulative consumption of group C was higher versus the other groups. Binge eating was observed in the ad libitum period of F1 and TF parents versus C and F2. There was no difference in the adiposity in the offspring.

**Conclusions:** Pregnant rats subjected to intermittent fasting were able to adjust energy homeostasis while maintaining adequate gestational weight gain, still the restriction induced metabolic programming and ruptured the hypothalamic anorectic response of adult offspring, increasing the risk of obesity at this stage.

**Keywords:** intermittent feeding, metabolic programming, female rat pregnancy, male rat offspring
# THE RELATIONSHIP OF NUTRITIONAL STATUS WITH INTELLECTUAL COEFFICIENT AND ACADEMIC ACHIEVEMENT IN MEXICAN CHILDREN FROM URBAN PRIMARY SCHOOLS

(1)Galván, Marcos; (2)Guerrero-Barreto, Esmeralda; (1)López-Rodríguez, Guadalupe; (1)Guzmán-Saldaña, Rebeca; (3)Suárez-Dieguez, Teodoro.

(1)Phd. Professor. U Autonoma de Hidalgo; (2)MsC. Nutritionist. U Autonoma de Hidalgo.; (3)MsC. Professor. U Autonoma de Hidalgo.

**Background and objectives:** The objective was to determine the magnitude of association of the nutritional status with the IQ and academic performance in children from urban primary schools in Hidalgo, Mexico.

**Methods:** An analytical observational study was carried out on 459 schoolchildren aged 6 to 10 years old from a public and a private school located in Tula, Hidalgo, during the 2013-2014 school cycle. The students were given weight and height measurements with calibrated instruments and standardized personnel, Koppitz test was applied to assess IQ and emotional indicators (EI), the grade point average was recorded as an indicator of academic achievement (AA), and socioeconomic status (SE) was assessed with household assets and father's level of schooling. Body mass index (BMIZ) and height-for-age (HAZ) Z scores were calculated using WHO 2007 references. Differences, correlations and interactions were assessed, and the magnitude of association of predictor variables (BMIZ and HAZ) and response (IQ and AA) with adjusted linear regression models (LRM).

**Results:** In school children of middle SE, obesity was 10 percent higher compared to high and low SE (p <0.01). The grade point average was higher in school children of high SE (8.9) compared to low SE (8.1) (p <0.01), and IQ average was higher in high SE school children (4.7) compared to low SE (4.2) (p <0.01). HAZ correlated positively with AA and IQ in low SE school children (r = 0.30 and r = 0.27, respectively); and the BMIZ correlated negatively with AA in school children of high SE (r = -0.41). The LRM for AA of school children of low SE, the HAZ, adjusted for EI, age and gender; explained the 33% of academic performance. The LRM for IQ of school children of low SE, the BMIZ, adjusted for EI, age and gender; explained the 44% of the IQ.

**Conclusions:** The anthropometric variables of HAZ and BMIZ explained the third part of academic and cognitive performance in low SE school children; indicating the strong influence that nutrition still has on the poorest children in countries in economic transition.

Financing. - This study was supported by CONACYT project no. 216092 and PRODEP UAEH-CA-86.

**Keywords:** Nutritional status, cognitive development, achievement performance, school children.

**Further collaborators:**

Fernanda Ríos-Pérez, Liliana López-Moctezuma.

# HEPATIC STEATOSIS INDUCED BY AMINO ACID DEFICIENCY OR BY MANIPULATION OF THE DIEDINARY AMINO ACID COMPOSITION

(1)Nishi, Hiroki; (2)Yamanaka, Daisuke; (3)Kamei, Hiroyasu; (1)Goda, Yuki; (4)Kumano, Mikako; (5)Toyoshima, Yuka; (6)Takenaka, Asako; (7)Chida, Kazuhiro; (7)Hakuno, Fumihiko; (7)Takahashi, Shin-Ichiro.

(1)MSc. Graduate School of Agricultural and Life Sciences. The University of Tokyo. Japan.; (2)PhD. Graduate School of Agricultural and Life Sciences. The University of Tokyo. Japan.; (3)PhD. Institute of Science and Engineering. Kanazawa University. Japan.; (4)BSc. Graduate School of Agricultural and Life Sciences. The University of Tokyo. Japan.; (5)PhD. Department of Bioregulation. Nippon Medical School. Japan.; (6)PhD. Department of Agricultural Chemistry. Meiji University. Japan.; (7)PhD. Graduate School of Agricultural and Life Sciences. The University of Tokyo. Japan.

**Background and objectives:** A low-protein diet causes young rats to decrease serum insulin and IGF-I levels, leading to mild growth retardation. In addition, such animals show increased insulin sensitivity and marked lipid accumulation in peripheral metabolic organs. In the present study, we unveiled this process to induce lipid accumulation in response to amino acid deficiency, especially focusing on hepatic steatosis development.

**Methods:** For animal experiments, we prepared a control diet containing 15% (w/w) amino acids equivalent to 15% casein and a low-amino acid diet containing 5% (w/w) amino acids or diets deficient in a single amino acid, and fed these diets to 6-week-old male Wistar rats for 7 days. Additionally, we applied a cell-culture model in which rat primary hepatocytes, and H4IIE and HuH7 cells, rat and human hepatoma cell lines respectively, were cultured in serum-free media containing complete amino acids (Full) or no amino acids (Zero).

**Results:** While a low-amino acid diet could induce triglyceride (TG) accumulation in the rat liver, simply a low-threonine or low-arginine diet could also develop the hepatic steatosis, accompanying an unexpected change in serum amino acid profiles, which implies that particular amino acid-specific signals were involved in hepatic lipid metabolic regulation. Since respiratory exchange ratios were not significantly different between control group and low-amino acid diet group, their lipid consumption rate might be comparable. On the other hand, when hepatocytes were cultured in Zero medium, their intracellular TG levels were increased, compared to Full medium, indicating that amino acid deficiency can induce hepatocytes to accumulate TG in a cell-autonomous manner. In addition, lipid synthesis activity of H4IIE and HuH7 cells using glucose as a substrate was enhanced in Zero medium. Similar to the rat model, when H4IIE cells were cultured in media deficient in a single amino acid, TG accumulation levels varied according to the amino acid composition of the media.

**Conclusions:** We conclude that a manipulation of dietary amino acid composition causes a dynamic change in blood amino acid profiles, which is sensed by hepatocytes and accelerates their lipid synthesis in a cell-autonomous manner, leading to hepatic steatosis.

**Keywords:** amino acid, liver, triglyceride, steatosis, rat
LYSINE DEFICIENCY SIGNAL SELECTIVELY INDUCES LIPID ACCUMULATION IN MUSCLE AND ADIPOSE TISSUES OF RATS

(1)Hakuno, Fumihiko; (2)Goda, Yuki; (2)Nishi, Hiroki; (2)Kumano, Mikako; (3)Yamanaka, Daisuke; (4)Yamanouchi, Keitaro; (4)Katsumata, Masaya; (5)Kato, Hisanori; (4)Nishihara, Masugi; (3)Takahashi, Shin Ichiro.

(1)PhD. Molecular Biologist. Graduate School of Agriculture and Lie Sciences. The University of Tokyo.; (2)Graduate Student. Nutritionist. Graduate School of Agricultural and Life Sciences. The University of Tokyo. Japan; (3)PhD. Endocrinologist. Graduate School of Agricultural and Life Sciences. The University of Tokyo. Japan; (4)PhD. Veterinary. Graduate School of Agricultural and Life Sciences. The University of Tokyo. Japan; (5)PhD. Nutritionist. Graduate School of Agricultural and Life Sciences. The University of Tokyo. Japan.

**Background and objectives:** We have previously reported that dietary amino acid restriction reduces serum levels of IGF-I and insulin, resulting in growth retardation, and additionally accumulation of triglycerides (TG) in liver, skeletal muscle and adipose tissues. The present study was undertaken to elucidate the effects of dietary amino-acid deprivation on lipid accumulation in muscle and adipose tissues.

**Methods:** We fed growing rats with the control diet containing sufficient (15%, w/w) amino acids (CN), or deficient (5%, w/w) amino acids (5AA) or diets deficient in each single amino acid in which only a single amino acid was reduced to the same amount as a 5AA diet and the others were the same as a CN diet (low AA) for a week. TG content was measured in muscle and liver tissue. And adipose tissues were imaged by computerized tomography (CT) scans.

**Results:** Lipid accumulated in adipose and skeletal muscle tissues in rats fed the 5AA diet and low-Lys diet compared with rats fed CN diets. Then rats were fed the 5AA diet or low-Lys diet for 8 weeks and we investigated lipid accumulation in each tissue. Adipose tissues in abdomen were imaged by CT scans and we calculated the volume of adipose tissues in each rat. The volume of adipose tissues was increased in rats fed the 5AA diet and low-Lys diets. The cell size of adipocytes of epididymis fat tissue was enlarged in the rats fed the 5AA and low-Lys diets. TG accumulation in skeletal muscle was also increased at 2-weeks feeding with 5AA and low-Lys diets.

**Conclusions:** These data suggest that depletion of lysine from the diets resulted in the accumulation of lipids in muscle and adipose tissue. Taken together with the data that lipid accumulated in liver in rats fed with low-Arg diets, selective amino acid deprivation from the diet could become a distinct signal to exert insulin-like activities for lipid metabolism, leading to induction of lipid accumulation in liver, skeletal muscle or adipose tissue selectively.

**Keywords:** amino acid, muscle, adipose tissue, lipid accumulation

EFFECTS OF DIETARY B VITAMINS INTAKE AND FOLATE METABOLISM IN THE DEVELOPMENT OF MILD COGNITIVE IMPAIRMENT AND THE POTENTIAL UNDERLYING MECHANISM

(1)An, Yu; (1)Zhang, Xiaona; (1)Lu, Yanhui; (1)Rong, Hongguo; (1)Tao, Lingwei; (2)Yu, Huanling; (2)Yuan, linhong; (2)Xiao, Rong.

(1)PhD. School of Public Health. Capital Medical University. China.; (2)PhD. Professor. School of Public Health. Capital Medical University. China.

**Background and objectives:** There is emerging evidence that blood levels of several B vitamins and cognitive function may be affected by dietary intake of these vitamins but the underlying mechanism is still controversial. DNA methylation together with oxidative stress are key to understanding the pathogenesis of mild cognitive impairment (MCI). The present study aimed to examine the relationship between B vitamins intake and MCI among the middle-older aged and elderly in China and shed light on the potential mechanism.

**Methods:** A cross-sectional study was carried out in 1487 Chinese adults aged 50 and above. A selection of comprehensive cognitive tests was administered to evaluate their cognitive performance. MCI patients were screened by using Mini-mental state examination (MMSE) Montreal Cognitive Assessment (MoCA) test and then diagnosed by neurologists. A semi-quantified Food frequency questionnaire (FFQ) method was used for dietary intake of B vitamins (mainly folic acid, vitamin B6, and vitamin B12) survey. Serum folate metabolism and oxidative stress markers were measured with ELISA. Global DNA methylation was determined by Illumina Methylation 450K chips.

**Results:** Subjects with higher intake of folate, vitamin B6 and vitamin B12 performed better in memory and processing speed. MCI patients had significantly higher concentrations of homocysteine (P<0.001) and oxidative stress markers(P<0.001) and lower folate (P<0.001) than non-MCI subjects. Serum levels of homocysteine was negatively correlated with folate and positively correlated with oxidative stress markers. Besides, 2277 different methylation gene loci were screened between MCI patients and non-MCI subjects, suggesting several gene function and metabolic pathways associated with MCI development.

**Conclusions:** Inadequate dietary intake of B vitamins, disturbances in folate and homocysteine metabolism and oxidative stress, with different DNA methylation as essential cofactors, are integral to MCI development.

**Keywords:** B vitamins, DNA methylation, folate metabolism, oxidative stress, mild cognitive impairment
# MECHANISM FOR THE DECREASE IN PLASMA ALPHA-TOCOPHEROL UNDER PROTEIN DEFICIENCY

(1)Takenaka, Asako; (2)Sato, Ryota; (3)Osada, Kyoichi; (4)Kawabata, Hiroaki.

(1)PhD. School of Agriculture. Meiji University. Japan; (2)Graduate School of Agriculture. Meiji University. Japan.; (3)School of Agriculture. Meiji University. Japan.; (4)School of Agriculture. Meiji University. Japan.

**Background and objectives:** Alpha-tocopherol is a major form of vitamin E and one of the most important antioxidants in mammals. It has been reported that plasma alpha-tocopherol concentration was reduced in rats fed protein deficient diet, which means that oxidative stress is increased under the condition of protein deficiency. In the present study, we investigated the mechanism for the decrease in plasma alpha-tocopherol concentration under protein deficiency in rats.

**Methods:** Five-week-old male Wistar rats were fed a control diet containing 20% casein (20C) or a low protein diet containing 5% casein (5C) for 7 days. Alpha-tocopherol concentrations in plasma, liver, peripheral tissues, feces and bile were measured. In order to investigate the effect of protein deficiency on alpha-tocopherol absorption in the intestine and transport from the liver into the circulation, mRNA and protein levels of intestinal transporters (NPC1L1 and SR-BI) and hepatic transporter (alpha-TTP) were measured. In addition, we fed rats with 20C or 5C with/without 0.08% ezetimibe, an inhibitor of NPC1L1, and measured plasma, liver, and fecal alpha-tocopherol concentrations to estimate the contribution of NPC1L1 on alpha-tocopherol absorption.

**Results:** Plasma and tissue alpha-tocopherol concentrations were lower in protein deficient rats while protein deficiency did not affected hepatic alpha-TTP mRNA and protein levels. These results demonstrate that reduced plasma and tissue alpha-tocopherol concentration is not caused by the decreased alpha-tocopherol transport activity from the liver into the circulation. On the other hand, fecal excretion of alpha-tocopherol was increased in protein deficient rats, demonstrating that reduced alpha-tocopherol absorption in the small intestine is a possible cause for the decrease in plasma and tissue alpha-tocopherol concentration in protein deficient rats. Ezetimibe did not affect fecal excretion of alpha-tocopherol and intestinal NPC1L1 level was not affected by protein deficiency, which demonstrated that reduced absorption of alpha-tocopherol was not caused by NPC1L1. On the other hand, intestinal SR-BI level was decreased in protein deficient rats that may reduce alpha-tocopherol absorption.

**Conclusions:** Protein deficiency reduces intestinal SR-BI and increases fecal excretion of alpha-tocopherol, which results in the reduction of plasma and tissue alpha-tocopherol concentrations in rats.

**Keywords:** alpha-tocopherol, protein deficiency, vitamin E

# MYOKINE RESPONSES TO RESISTANCE EXERCISE WITH DIFFERENT NUTRIENT AVAILABILITY ON A CONCURRENT EXERCISE DAY IN YOUNG, HEALTHY, PHYSICAL ACTIVE MALES

(1)Mensink, Marco; (2)Knuiman, Pim; (3)Hopman, Maria;

(2)Hangelbroek, Roland.

(1)PhD. MD. Assistant Professor. Division of Human Nutrition. Wageningen University. The Netherlands; (2)MSc. PhD-student. Division of Human Nutrition. Wageningen University. Wageningen. The Netherlands; (3)PhD. Professor. Department of Physiology. Radboud University Medical Centre. Nijmegen. The Netherlands.

**Background and objectives:** Myokines are proteins produced and released by skeletal muscle in response to exercise, and suggested to play a role in exercise-induced adaptations of muscle tissue. Nutrient availability, in particular carbohydrate (CHO), is proposed as a potential regulator of myokine responses. We evaluated the effects of a pre-exercise carbohydrate-rich meal versus a low carbohydrate -high fat meal on plasma myokine responses to resistance exercise after a glycogen-depleting endurance exercise earlier that day.

**Methods:** Fifteen males performed two experimental days with a 90 min bout of endurance exercise at 70% VO2max in the morning (8.30-10 am) and resistance exercise in the afternoon (2-2.30 pm):5 x 8 80% 1-RM repetitions of bilateral leg press and extension. Either a high CHO/low fat meal (110g CHO, 52g protein, 9g fat; ~750 kcal) or a iso-caloric low-CHO/high fat meal (20 gr CHO, 52g protein, 51g fat) was provided 2h before resistance exercise (noon). Blood was drawn after an overnight fast and at regular time intervals up to 3 hours after resistance exercise to measure plasma myokine/cytokine levels.

**Results:** Postprandial plasma glucose and insulin levels were higher, and FFA lower, on the high-CHO condition compared to the low-CHO. IL-6 increased post-exercise, and was significantly higher after the low-CHO meal in the post-resistance exercise period compared to the high-CHO meal. IL-8 and IL-10 were only increased after the endurance exercise. IL-15 increased postprandial on the high-CHO condition only, and was increased in the early post-resistance exercise period, with a slightly higher level on the high-CHO condition. TNFα and Decorin did not show a clear response, while ANGPTL4 was slightly elevated post-exercise, and strongly increased in the postprandial period, with no differences between meals. MCP-1 tended, on both occasions, to increase after the endurance exercise with subsequent a reduction below baseline.

**Conclusions:** The composition of the pre-exercise meal did in general not influence myokine responses in the post-resistance exercise period, although plasma IL-6 levels were higher in the low-CHO condition compared with high-CHO. Our findings support the view that pre-resistance exercise carbohydrate availability does not have a major impact on acute responses of myokines.

**Keywords:** Resistance Exercise, carbohydrate availability, myokines, Interleukin-6
# EFFECT OF EXERCISE STIMULUS-INDUCED ANDROGEN STEROIDOGENESIS ON LCFA TRANSPORT PROTEINS IN SKELETAL MUSCLE

(1)Kim, Jisu; (2)Park, Jonghoon; (3)Nahyun, Kim; (4)Kim, Eunseok; (4)Park, Yeram; (5)Lim, Kiwon.

(1)Physical activity & Performance Institute. Konkuk University. Korea; (2)Ph.D. professor. Department of Physical Education. Korea University. Seoul. Korea; (3)Department of Physical Education. Korea University. Seoul. Korea; (4)Department of Physical Education. Konkuk University. Seoul. Korea; (5)Ph.D. professor. Department of Physical Education. Konkuk University. Seoul. Korea.

**Background and objectives:** The aim of this study was to determine whether an acute of aerobic exercise would increase androgen hormone concentration in skeletal muscle and LCFA transport proteins.

**Methods:** 20 Male ICR mice, 9 weeks old, were randomized into three groups: Control (n=7, CON), Exercise (n=7, EX), and Exercise + SRD5A1A2 Inhibitor (n=7, EX-IN, SRD5A1A2 is an enzyme to be needed when free testosterone (FT) is metabolized to dihydrotestosterone (DHT)). Inhibitor was administered to the EX-IN intraperitoneally while the CON and EX were treated with vehicle only. After 1-hour administration in mice, exercise was performed at 60-70%VO2max for 30min. Respiratory Exchange Ratio (RER) during exercise were measured in calorimeter. The levels of FT and DHT in Skeletal muscle tissue were determined using an ELISA kit, and FAT/CD36 and CPT1 were examined by western blot.

**Results:** We found that FT and DHT concentration in skeletal muscle were no significantly different among the groups. However, EX group had significantly lower integrated area under the curve in RER at last 10-min during an exercise than EX-IN. The EX and EX-IN groups had greater FAT/CD36 expression in skeletal muscle than CON group. Similarly, EX groups had significantly higher CPT1 expression than in CON group, whereas EX-IN group had significantly lower than in EX group.

**Conclusions:** Our results provide a novel finding that aerobic exercise did not change FT and DHT levels in skeletal muscle. However, androgen metabolism on skeletal muscle in association with the increase of CPT1 gene activation.

**Keywords:** Androgenmetabolism, LCFA transport protein, Fat oxidation, Energy metabolism during exercise

# CARBON STABLE ISOTOPE RATIOS HAVE POTENTIAL AS A BIOMARKER FOR THE ASSESSMENT OF SUGARS INTAKE IN NEW ZEALAND POPULATIONS

(1)Kruimer, Devonia; (2)Te Morenga, Lisa; (3)Mann, Jim; (4)Merriman, Tony.

(1)MSc. PhD Candidate. Department of Human Nutrition and Riddet Institute. University of Otago. New Zealand; (2)PhD. Senior Research Fellow. Department of Human Nutrition. Edgar Diabetes and Obesity Centre and Riddet Institute. University of Otago. New Zealand; (3)PhD. Professor. Departments of Human Nutrition and Medicine. Edgar Diabetes and Obesity Centre and Riddet Institute. University of Otago. New Zealand; (4)PhD. Professor. Department of Biochemistry and Edgar Diabetes and Obesity Centre. University of Otago. New Zealand.

**Background and objectives:** Carbon stable isotope ratios (13C/12C; expressed as δ13C) in various tissues have recently been shown to predict intakes of free sugars derived from cane and corn in U.S. populations. Whereas corn, cane and sugar-beet derived sugars are commonly used in the U.S., cane sugar predominates in New Zealand (NZ). Our objective was to investigate whether δ13C is associated with sugars intakes in two small cohort studies conducted in NZ.

**Methods:** The first cohort included young people who identified with at least one Pacific Island group aged 15-20y (n = 80). The second involved Māori (indigenous) adults living in a semi-rural community on the East Coast of NZ (n= 37). Sugars intakes were assessed using culturally tailored and validated food frequency questionnaires. Isotopic compositions for carbon (δ13C) and nitrogen (δ15N) in red blood cells were determined by combustion in an Elemental Analyser and measurement of the resulting N2 and CO2 gases by Isotopic Ratio Mass Spectrometry (EA-IRMS). Compound-specific analysis of the stable isotope ratios in the alamine fraction (δ13Calanine) of red blood cells was determined by Gas Chromatography with combustion and Ipe Ratio Mass Spectrometry (CG-IRMS).

**Results:** In the Pacific cohort a dual isotope model including δ13C and δ15N correlated with self-reported free sugars intakes; r=0.48 for total free-sugars and r=0.61 for sugar-sweetened beverage intakes in normal-weight subjects (body mass index (BMI): 19-25 kg/m2), but not in overweight and obese participants. Conversely, δ13Calanine was not correlated with free sugars intakes. In the Māori cohort δ13Calanine was correlated with total sugar intake (r= 0.49, p=0.002), sucrose intake (r=0.45, p=0.004), and added sugars intake (r=0.49, p=0.002). Linear regression analysis adjusted for sex, age, BMI and HbA1c showed that δ13Calanine was strongly associated with total sugars intake (r2= 0.49, p = 0.002). The dual isotope model including δ13C and δ15N was not associated with self-reported sugar intakes.

**Conclusions:** Carbon stable isotope ratios in red blood cells have potential as objective biomarkers of sugar intakes in New Zealand population studies. A larger validation study involving optimal dietary assessment method is now warranted to confirm these preliminary findings.
EFFECT OF THE PPM1K GENETIC VARIANT ON
GLUCOSE METABOLISM TRAITS IN RESPONSE
TO WEIGHT LOSS DIETS

(1)MSc. Department of Nutrition, Food Sciences and Physiology. Centre for Nutrition Research. University of Navarra. Spain/ CIBERobn. ISCIII. Spain/ Navarra Institute for Health Research (IdiSNA). Spain; (2)MD. PhD. Department of Epidemiology. Tulane University. New Orleans / Department of Epidemiology and Department of Nutrition. Chan School of Public Health. Boston. USA; (3)PhD. Department of Nutrition, Food Sciences and Physiology. Centre for Nutrition Research. University of Navarra. Spain/ CIBERobn. ISCIII. Spain/ Navarra Institute for Health Research (IdiSNA). Spain; (4)MD. Novo Nordisk Foundation Center for Basic Metabolic Research. University of Copenhagen. Denmark / Danish Diabetes Academy. Odense. Denmark; (5)MD. PhD. Novo Nordisk Foundation Center for Basic Metabolic Research. University of Copenhagen. Denmark; (6)MD. PhD. Department of Nutrition, Food Sciences and Physiology. Centre for Nutrition Research. University of Navarra. Spain/ CIBERobn. ISCIII. Spain/ Navarra Institute for Health Research (IdiSNA). Spain.

**Background and objectives:** Circulating branched chain amino acids (BCAAs) have been related to insulin resistance and type 2 diabetes (T2D) risk. We tested whether the BCAAs-increasing rs1440581 PPM1K T-allele affected changes in glucose metabolism traits depending on dietary fat intake during a weight loss dietary intervention.

**Methods:** The rs1440581 PPM1K variant was genotyped in 757 non-diabetic individuals (NUGENOB study). Participants were randomly assigned to one of the two energy restricted diets (low-fat diet: 20-25% fat, 15% protein, 60-65% carbohydrate; high-fat diet: 40-45% fat, 15% protein and 40-45% carbohydrate). The changes in fasting glucose, insulin, insulin resistance (HOMA-IR) and cell function (HOMA-B) were measured after 10 weeks of intervention.

**Results:** Significant interactions were found between the PPM1K genotype and low-/high-fat diet on changes in insulin, HOMA-IR and HOMA-B (p interaction 0.01, 0.02 and 0.003, respectively). In the low-fat dietary group the T allele was marginally associated with smaller decreases in insulin and HOMA-IR (0.69 U/mL per risk allele, p=0.07; and 0.21 per risk allele, p=0.04; respectively). On the other hand, in the high-fat diet group, the T allele was associated with decrease in HOMA-B ( -12.5 per risk allele p=0.006).

**Conclusions:** Our findings indicate that the rs1440581 polymorphism may differently influence changes in glucose metabolism during weight loss depending on dietary fat intake.

EXTRA-VIRGIN OLIVE OIL OR NUTS CONSUMPTION AND DNA METHYLATION IN PERIPHERAL BLOOD CELLS WITHIN THE PREDIMED-NAVARRA TRIAL

(1)MSc. Department of Nutrition, Food Sciences and Physiology. Centre for Nutrition Research. University of Navarra. Spain;
(2)PhD. Department of Nutrition, Food Sciences and Physiology. Centre for Nutrition Research. University of Navarra. Spain/
CIBERobn. ISCIII. Spain; (3)PhD. Department of Nutrition, Food Sciences and Physiology. University of Navarra. Spain/
CIBERobn. ISCIII. Spain/ Navarra Institute for Health Research (IdiSNA).
Spain; (4)PhD. Department of Preventive Medicine and Public Health. University of Navarra. Spain.; (5)PhD. Department of
Nutrition, Food Sciences and Physiology. Centre for Nutrition Research. University of Navarra. Spain/ Navarra Institute for
Health Research (IdiSNA). Spain; (6)MD. PhD. Department of Nutrition, Food Sciences and Physiology. Centre for Nutrition
Research. University of Navarra. Spain/ CIBERobn. ISCIII. Spain/
Navarra Institute for Health Research (IdiSNA). Spain.

**Background and objectives:** Epigenetic marks, including DNA methylation, might be reversible and modulated by environmental factors such as dietary exposures. Some dietary components have demonstrated to mediate metabolic effects through methylation processes. Therefore, some of the beneficial effects of the Mediterranean diet (MedDiet) could involve methylation changes. The objective of this study was to analyse whether an intervention with a MedDiet supplemented with extra virgin olive oil (MedDiet+EVOO) or nuts (MedDiet+nuts) was able to influence the methylation levels in peripheral blood cells.

**Methods:** A subset of 36 individuals was selected within the PREvención con DIeta MEDITerránea (PREDIMED)-NAVARRA study, a randomised, controlled, parallel trial with three groups of intervention in high cardiovascular risk volunteers: MedDiet+EVOO, MedDiet+nuts and a low-fat control group. Methylation levels were measured at 5 years of intervention and at baseline. CpG sites (CpGs) with high changes in methylation and homogeneous dispersion with putative biological implications were chosen by statistically restricted selections.

**Results:** Five CpGs related to intermediate metabolism, diabetes, obesity, inflammation and signal transduction were finally selected; one with methylation differences in the MedDiet+nuts group (cg01081346-CPT1B) and four in the MedDiet+EVOO group (cg00173776-PUM1, cg12147622-DDIT4, cg21870545-EPB41, cg24045357-LINC01117).

**Conclusions:** Specific components of MedDiet, particularly nuts and EVOO, were able to induce changes in methylation levels

<footnote>(1)Goni, Leticia; (2)Qi, Lu; (3)Cuervo, Marta; (3)Milagro, Fermin I.;</footnote>

<footnote>(4)Svendstrup, Mathilde; (5)Sørensen, Thorkild I.A.; (5)Hansen, Torben;</footnote>

<footnote>(6)Martínez, J. Alfredo.</footnote>

<footnote>(1)Arpón, Ana; (2)Milagro, Fermin I.; (3)Marti, Amelia; (4)Ruiz-Canela, Miguel; (5)Riezu Boj, José Ignacio; (6)Martínez, J. Alfredo.</footnote>
in several genes from peripheral blood cells. These methylation
changes may be related to some of the potential benefits in health
and may contribute to explain the role of MedDiet on health out-
comes.

**Keywords:** Epigenetics, DNA methylation, Mediterranean diet, Olive oil, Nuts

**Further collaborators:**

PREDIMED investigators

low folic acid was related to salt inducible kinase 1 (SIK1) gene
hypomethylation. Moreover, methylation levels of SIK1, a direct
CREB target gene involved in glucose metabolism, negatively cor-
related with HOMA-IR index.

**Conclusions:** These results suggest that SIK1 hypomethylation could be an epigenetic mechanism underlying low folic acid-induced insulin resistance in obese subjects.

**Keywords:** Folic acid, epigenetics, DNA methylation, obesity, insulin resistance

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LOW FOLIC ACID INTAKE IS ASSOCIATED WITH SIK1 HYPOMETHYLATION AND INSULIN RESISTANCE IN OBESE SUBJECTS

(1)Ramos-Lopez, Omar; (2)Samblas, Mirian; (3)Milagro, Fermin I.;

(1)Mansego, María L.; (4)Zulet, Maria Angeles; (5)Riezu Boj, José Ignacio; (6)Martínez, J. Alfredo.

(1)PhD. Department of Nutrition, Food Sciences and Physiology.
Centre for Nutrition Research. University of Navarra. Spain;

(2)MSc. Department of Nutrition, Food Sciences and Physiology.
Centre for Nutrition Research. University of Navarra. Spain; (3)PhD.
Department of Nutrition, Food Sciences and Physiology. Centre
for Nutrition Research. University of Navarra. Spain / CIBERobn.
ISCIII. Spain; (4)PhD. Department of Nutrition, Food Sciences and
Physiology. Centre for Nutrition Research. University of Navarra.
Spain / CIBERobn. ISCIII. Spain / Navarra Institute for Health

Research (IdiSNA). Spain; (5)PhD. Department of Nutrition, Food
Sciences and Physiology and Centre for Nutrition Research.
University of Navarra. Spain / Navarra Institute for Health Research

(IdiSNA). Spain; (6)MD. PhD. Department of Nutrition, Food
Sciences and Physiology. Centre for Nutrition Research. University
of Navarra. Spain / CIBERobn. ISCIII. Spain / Navarra Institute for

Health Research (IdiSNA). Spain.

**Background and objectives:** Folic acid is involved in the regulation of many biological processes including DNA methylation via the one-carbon metabolism pathway. Furthermore, folic acid deficiency has been putatively implicated in the onset of diverse metabolic diseases, including insulin resistance, by altering DNA methylation patterns on key regulatory genes. The aim of this study was to investigate the association between folic acid intake and metabolic features, with emphasis on gene-specific DNA methylation patterns.

**Methods:** A cross-sectional ancillary study was conducted in obese subjects (n=47) from the RESMENA study (Spain). Fat mass was measured by dual-energy X-ray absorptiometry (DXA). Dietary intake and the metabolic profile were assessed by standardized methods. DNA methylation in peripheral white blood cells was analyzed by microarray (Infinium Human Methylation 450K BeadChips).

**Results:** Subjects with a folic acid intake lower than 300 µg/day showed more fat mass (especially trunk fat), as well as higher levels of glucose, insulin, HOMA-IR index, cortisol and PAI-1 than those consuming at least or more than 300 µg/day. Noteworthy,

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IMPROVEMENTS IN INSULIN RESISTANCE AND FGF21 IN OBESE SUBJECTS AFTER A LONG-TERM WEIGHT LOSS INTERVENTION: RESMENA PROJECT

(1)Cantero González, Irene; (2)Abete, Itziar; (3)Crujeiras, Ana Belén;

(4)Casanueva, Felipe F; (5)Martínez, J. Alfredo; (6)Zulet, Maria Angeles.

(1)MSc. Department of Nutrition, Food Sciences and Physiology.
Centre for Nutrition Research. University of Navarra. Spain; (2)PhD.
Department of Nutrition, Food Sciences and Physiology. Centre
for Nutrition Research. University of Navarra. Spain/ CIBERobn.
ISCIII. Spain.; (3)PhD. CIBERobn / Division of Endocrinology,
Department of Medicine, Complejo Hospitalario Universitario
de Santiago (CHUS/SERGAS), Instituto de Investigación Sanitaria
de Santiago (IDIS), Santiago de Compostela, Spain; (4)MD, PhD.
CIBERobn / Division of Endocrinology, Department of Medicine,
Complejo Hospitalario Universitario de Santiago (CHUS/SERGAS),

Instituto de Investigación Sanitaria de Santiago (IDIS), Santiago de Compostela, Spain; (5)MD, PhD. Department of Nutrition, Food Sciences and Physiology. Centre for Nutrition Research. University of Navarra. Spain/ CIBERobn. ISCIII. Spain/ Navarra Institute for Health Research (IdiSNA). Spain; (6)PhD. Department of Nutrition, Food Sciences and Physiology. Centre for Nutrition Research. University of Navarra. Spain/ CIBERobn. ISCIII. Spain/ Navarra Institute for Health Research (IdiSNA). Spain.

**Background and objectives:** Fibroblast growth factor 21 (FGF-21), a potent activator of glucose uptake, has been proposed to be related to insulin resistance, metabolic syndrome (MetS), nonalcoholic fatty liver disease (NAFLD), and weight status. The objective was to evaluate the relationship between FGF21 and criteria of MetS in obese subjects after 6 months under energy restricted diet.

**Methods:** Sixty-six adults (55.5% men; 49 ± 9 years; BMI: 35.7 ± 4 kg/m2) of RESMENA study were evaluated at baseline and after 6 months. Blood pressure, anthropometric, body composition (DXA), glycemic and lipid profile (Pentra C-200) as well as FGF21 (ELISA assay) were determined.

**Results:** The participants showed a significant reduction in weight (6.9 ± 2.9 kg), and waist circumference, body fat, visceral adipose tissue, blood pressure, total cholesterol, LDL-c, TG, glucose and HOMA-IR. Interestingly, FGF21 significantly decreased after weight loss intervention (p<0.001), whose change positively
and significantly correlated with variation in HOMA-index (r= 0.312; p= 0.015). Thus, a linear regression model was set up to assess the influence of FGF21 on HOMA-index (β= 23.6; R2= 0.112; p= 0.040) revealing that the association depend on weight loss.

**Conclusions:** Our data show a positive relationship between FGF21 and insulin resistance suggesting the potential mediating role of FGF21 on glucose homeostasis in weight loss.

**Keywords:** FGF21, obesity, weight loss, insulin resistance.

**Further collaborators:**

de la Iglesia R, Lopez-Legarrea P, Perez-Cornago A, Perez S, Martinez de Morentin BE, Ciaurriz V, Amil M.

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ELEVATED THYROGLOBULIN CONCENTRATIONS, BUT NO INCREASED PREVALENCE OF THYROID FUNCTION DISORDERS IN AREAS WITH CHRONIC EXCESSIVE IODINE INTAKE

(1)Farebrother, Jessica; (2)Untoro, Juliawati; (3)Assey, Vincent; (4)Makokha, Anselimo; (5)Andersson, Maria; (6)Zimmermann, Michael Bruce.

(1)MPharm MPH. Human Nutrition Laboratory. Institute of Food, Nutrition and Health. ETH Zurich. Switzerland.; (2)PhD. East and Southern Africa Regional Office (ESARO). UNICEF. Kenya;
(3)PhD. Nutrition Services. Ministry of Health and Social Welfare. Tanzania.; (4)PhD. Department of Food Science and Technology. Jomo Kenyatta University of Agriculture and Technology. Kenya.;
(5)PhD. Human Nutrition Laboratory. Institute of Food, Nutrition and Health. ETH Zurich. Switzerland.; (6)MD. Human Nutrition Laboratory. Institute of Food, Nutrition and Health. ETH Zurich. Switzerland.

**Background and objectives:** Excessive iodine intake may have adverse effects on thyroid function, including hyperthyroidism, hypothyroidism and thyroid autoimmunity, but the relative effects across the life cycle are unclear. Large regional variations in iodine intake have been reported in Eastern Africa, from deficient to well above recommended levels, questioning the adherence to existing universal salt iodization (USI) programmes. We sought to evaluate the effects of prolonged excessive iodine intake on thyroid function.

**Methods:** In this cross-sectional study, we evaluated the iodine status (UIC) and thyroid function (tT4, TSH, Tg on dried blood spots) of approximately 3600 participants from 6 population groups (pregnant, lactating and women of reproductive age, schoolchildren, breastfeeding and weaning infants), at two purposefully chosen sites in Kenya (K) and Tanzania (TZ) with previously documented excessive iodine intakes in schoolchildren. We analysed household salt iodine content and groundwater concentrations, and a questionnaire assessed the intake of iodised salt and consumption of foods rich in iodine.

**Results:** Iodine intake greatly exceeded minimum requirements in all groups: median UIC in schoolchildren was above the 300 g/L WHO cut-off indicating excessive intake, however below that of 500 g/L proposed during pregnancy. At 43.5 mg/kg (IQR 33.5, 54.0), salt iodine concentrations complied with national legislation at both sites.

Drinking water iodine concentrations ranged from 0 to 2943 g/L. The prevalence of thyroid disorders was low in all population groups (<0.5% overt, <2.8% subclinical disease), however a high prevalence of elevated thyroglobulin was observed in schoolchildren (K: 46.2%, TZ: 31.1%) and pregnant women (K: 20.1%; TZ: 16.7%).

**Conclusions:** In these populations, exposed to a prolonged iodine intake exceeding the dietary requirements, the prevalence of thyroid disorders was similar to iodine-sufficient populations suggesting that the thyroid gland may adapt to high intakes. The well-functioning USI programmes should be maintained, but monitoring and surveillance must be strengthened. Groundwater likely contributes to the high iodine intakes in the present study populations. In areas of known iodine excess, monitoring of groundwater sources should be encouraged.

**Keywords:** Iodine excess, thyroid function, thyroglobulin, USI, groundwater

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FEASIBILITY OF COLLECTING NUTRITION INFORMATION USING COMPUTER ASSISTED TELEPHONE INTERVIEWING

(1)Hachethu, Kusum; (2)Chesterman, Sabrina; (3)Mwongela, Beatrice;

(4)Ng'endo, Mary; (5)Bauer, Jean-Martin.

(1)MSc. Nutritionist.United Nations World Food Programme. Vulnerability Analyis and Mapping. Rome. Italy; (2)MSc. World Agroforestry Centre (ICRAF). Nairobi. Kenya; (3)Msc. United Nations World Food Programme. Nairobi. Kenya; (4)PhD. World Agroforestry Centre (ICRAF). Nairobi. Kenya; (5)MPP. United Nations World Food Programme. Rome. Italy.

**Background and objectives:** Regular information on nutrition status within target populations is needed to monitor changes over time, provide early warning, and guide policies and programmes linked to nutrition in vulnerable counties. With rapid growth in mobile phone access and ownership in sub-Saharan Africa, data collection using mobile methodologies such as Computer-Assisted Telephone Interviewing (CATI) could enable quick, affordable and high frequency data collection at scale.

The objective of the study was to understand mobile phone access, ownership, and usage patterns by rural women in Kenya, in particular the opportunities and constraints for monitoring nutrition via mobile phone interviews with women of reproductive age and caregivers of young children.

**Methods:** We conducted 17 focus group discussions and 16 in-depth interviews with women, as well as 22 key informant interviews with mobile phone vendors, local nutritionists and local government representatives. The study was conducted in two counties in Kenya (Kitui and Baringo) representing different socioeconomic and agroecological conditions. Qualitative data were coded and analysed using NVivo 10 based on defined codes; additional codes and sub-nodes were added based on emerging themes from the data.
**Results:** Our results show that the majority of women in both counties had access to phones through inter- and intra-household sharing, even if they didn't own one themselves. Similarly, a majority of participants showed high willingness to participate in mobile phone surveys related to diet. However, potential barriers for women to participate in mobile phone surveys included poor network coverage, trust issues, and gender norms.

**Conclusions:** Innovative data collection methodologies such as CATI can be used to collect nutrition information from rural women. Potential barriers related to trust issues and gender norms can be addressed through prior community sensitization, particularly with local leaders and male household heads. Mobile phone surveys are most feasible in areas where network coverage is good, however pre-scheduling of phone calls can minimize non-response in areas of poor network.

**Keywords:** Nutrition, Mobile survey, CATI, Data Innovation, Rural Kenya

**Conflict of Interest Disclosure:** United Nation World Food Programme's mobile Vulnerability Analysis and Mapping (mVAM) project uses various mobile methodologies to collect food security data.

## CHANGES IN OBESITY METABOLOMIC PROFILE ASSOCIATED WITH MANGANESE INTAKE IN A POPULATION BASED STUDY

(1)Macedo Silva, Alexsandro; (2)Mendes Gorgulho, Bartira; (3)Carioca, Antonio Augusto; (4)Fisberg, Regina; (5)Marchioni, Dirce.

(1)PhD Student. Department of Nutrition of University of Sao Paulo; (2)PhD. Nutricionist. Department of Nutrition. University of Sao Paulo. Brazil; (3)PhD Student. Department of Nutrition. University of Sao Paulo; (4)Professor. Nutritionist. Department of Nutrition. University of Sao Paulo. Brazil.; (5)Professor. Nutricionist. Department of Nutrition. University of Sao Paulo.

**Background and objectives:** The amino acids (AA) profile has been reported to be related to metabolism alteration, impairing the mitochondrial activity, in obese individuals. Manganese dietary intake inadequacy could collaborate in abnormal accumulation of body and tissue fat, such as impairers lipid metabolism, and glucose tolerance. The relationship between manganese intake, and AA profile could indicate mitochondrial metabolism alteration, and future diseases. The aim of this study was evaluate the association between dietary manganese intake and AA profile.

**Methods:** The dietary intake data were obtained from the "Health Survey of São Paulo" (ISA-Capital 2008), a population based cross-sectional survey in São Paulo, Brazil. The blood sample was obtained from 168 adults (≥20 years) and 21 amino acids in plasma were analyzed, using Affinity/HPLC with electrochemical detection method. AA profile was performed using factor analysis with extraction of principal components by varimax rotation. The factor loading considered the eigenvalue more than 0.4. Finally, linear regression was used to verify the relationship

between manganese intake and AA profile, adjusting by age, sex, and energy intake.

**Results:** The factor analysis retained five factors with cumulative variance of 0.662, and Kaiser-Meyer-Olkin of sampling adequacy equal to 0.824: F1 (Isoleucine, Leucine, Lysine, Methionine, Phenylalanine, Tryptophan, Tyrosine, Valine); F2 (Asparagine, Glycine, Serine, Threonine); F3 (Arginine, Glutamate, Histidine); F4 (Citrulline, Glutamine, Ornithine); F5 (Alanine, Aspartate, Proline). F1 (β= -0.2616; p=0.009) and F3 (β= 0.2047908, p=0.047) revealed significant association with the manganese intake. The F1, that represented the AA profile related to β-oxidation, was negatively correlated with manganese intake; while F3, that represent the citric acid cycle, was positively correlated with manganese intake.

**Conclusions:** The manganese intake was associate negatively to the AA profile related to the β-oxidation, composed mainly by the branch chain amino acids and aromatic amino acids, which ones have been associated to obesity, and metabolism alterations, causing future diseases. However, the manganese consumption revealed positively association with citric acid cycle AA profile, whereas the manganese might be related to the enzymes responsible for aerobic metabolism.

**Keywords:** Manganese Intake. Metabolomics. Obesity. Metabolism.

## L-LACTATE ADMINISTRATION STIMULATES FAT OXIDATION AND GLYCOGEN STORAGE IN RESTING RATS

(1)Yoo, ChoongSung; (2)Kim, Jisu; (3)Hashimoto, Takeshi; (4)Tomi, Hironori; (4)Kawasaki, Mihoko; (5)Teramoto, Noboru; (2)Park, Mi-Young; (6)Lim, Insoo; (7)Lim, Kiwon.

(1)BSc. Physical Education. Graduate School of Konkuk University. Republic of Korea.; (2)PhD. Physical Activity & Performance Institute. Konkuk University. Republic of Korea; (3)Department of Sport and Health Science. Ritsumeikan University. Japan; (4)Department of Central R&D Laboratory. Kobayashi Pharmaceutical Co., Ltd. Japan.; (5)researcher. OSU Co., Ltd. Japan; (6)PhD. Physical Education. Graduate School of Changwon University. Republic of Korea.; (7)PhD. Physical Education. Graduate School of Konkuk University. Republic of Korea.

**Background and objectives:** L-lactate is known as the end-product of glycolysis. Therefore it is considered as a fatigue product. However, since last decades there has been demonstrated positive effect of L-lactate. Nevertheless it is not known whether the L-lactate changes on energy metabolism and energy substrates utilization.

**Methods:** Study1. We investigated the effects of L-lactate treatment by using an open calorimetric chamber on resting rats for 6 hours. We randomized 32, 7-week-old male SD rats into 4 groups: the control (Con: DW), caffeine (Caf: 1mg/100g), L-lactate (Lac:400mg/100g), and caffeine and L-lactate mixed compound
(Caf+Lac: 1mg, 400mg/100g). We treated different substances to each groups by using sonde for oral administration. We examined 6 hours on energy metabolism in resting conditions by using respiratory metabolic chambers.

Study 2. We did Additional experiment to see gene movements within 2 hours with Caf+Lac group, because they were the highest group in fat oxidation. We treated same dose of Caf+Lac and sacrificed 30minutes, 60 minutes and 2 hours after the treatment. We sacrificed zero-point without any treatment and analyzed all the groups by RT PCR.

**Results:** Study1. There was no significant difference in oxygen uptake and carbon dioxide production between groups. However with fat oxidation, Lac and Caf+Lac were significantly higher than Con in first 2 hours after administration in fat oxidation.

Study2. MCT1 decreased after 2hours compared to zero and FAT/CD36 showed up and down tendency with significant difference within 2 hours. Finally PDK4 increased at 2hours compared to zero.

Also when we examined blood sample we could see there is a significant increase in glycerol level after 2hours.

Furthermore when we analyzed glycogen level in liver, we could find significant increase within 2 hours.

**Conclusions:** The rejection of glycolysis which might be understood as increase in glycogen synthesis, has possibility of shown to as increase in fat oxidation. For this reason, L-lactate administration could be an effective supplement for aerobic athletes' glycogen loading and therefore it needs to be studied more with metabolism clarification and dose response.

**Keywords:** L-lactate, Fat oxidation, Glycogen, Resting metabolic rate

144/1721

ANTHROPOMETRIC MEASUREMENTS ASSOCIATED TO METABOLIC SYNDROME AND INSULIN RESISTANCE IN CHILEAN SCHOOL AGE CHILDREN

(¹)Fernandez, Ximena; (²)Arnaiz, Pilar; (³)Barja, Salesa; (⁴)Villarroel, Luis; (⁵)Mardones, Francisco.

(¹)BSc. MSc. Department of Public Health. Pontificia Universidad Católica de Chile. Chile.; (²)MD. Department of Pediatrics. Pontificia Universidad Católica de Chile. Chile.; (³)MD. MSc. Department of Pediatrics. Pontificia Universidad Católica de Chile. Chile.; (⁴)PhD. Department of Public Health. Pontificia Universidad Católica de Chile. Chile.; (⁵)MD. MSc. Department of Public Health. Pontificia Universidad Católica de Chile. Chile.

**Background and objectives:** The World Health Organization has recommended determining - sex and population - specific relationships between measures of fatness and morbidity. Obesity anthropometric indicators were associated in this study of Chilean children with the metabolic syndrome (MS) and with insulin resistance (IR).

**Methods:** A cross-sectional study enrolled 3325 subjects attending 5th and 6th grade of primary education during 2009-2011. The relationship of MS and IR with anthropometric indicators was

assessed. Receiver operator characteristic (ROC) curves were used to compare discrimination and estimate optimal cutoffs of body mass index (BMI), fat mass (FM), waist circumference (WC) and waist-to-height ratio (WHtR). Sensitivity, specificity and net classification improvement (NRI) for these cut cutoffs were compared with values obtained applying international recommendations.

**Results:** Children were 52.2% females and mean age was 11.4±1 years. All anthropometric indicators had similar ROC areas for MS and RI, both in girls and boys. Optimal cutoffs slightly improved NR when compared with international recommendations.

**Conclusions:** The similarities observed permitted to propose two simpler anthropometric measures for clinical practice. They are BMI and WHtR; the latter has also the advantage of not needing tables to assess the specific diagnosis.

**Keywords:** children, insulin resistance, metabolic syndrome, fat mass, body mass index

**Conflict of Interest Disclosure:** This study was funded by the Chilean fund for science of technology (FONDECYT project No. 1090594) which had no influence in the presented results.

144/1723

EFFECT OF LEAA SUPPLEMENTATION ON MUSCULAR FATIGUE AND INFLAMMATORY CYTOKINES IN WHEELCHAIR BASKETBALL PLAYERS

(¹)An, Young Hwan; (²)Kim, Hee-Jae; (³)Jung, Yong Jin; (⁴)Fujimura, Naoko; (⁴)Toyoda, Takuya; (⁵)Yamada, Toshiyuki; (⁵)Uchida, Hirohisa;

(¹)Lim, Kiwon.

(¹)Department of Physical Education. Konkuk University. Seoul. Korea; (²)Physical Activity and Performance Institute (PAPI). Konkuk University. Seoul. Korea; (³)Department of Internal Medicine. Seoul National University. College of Medicine. SMG-SNU Boramae Medical Center. Seoul. Korea; (⁴)Institute of Food Science & Technologies. Ajinomoto Co., Inc.. Kanagawa. Japan;

(⁵)Sports Nutrition Department. Ajinomoto Co., Inc.. Tokyo. Japan.

**Background and objectives:** Several studies found that an effect of amino acid supplement on reducing muscle damage and attenuated development of delayed onset muscle soreness (DOMS) induced by exercise. This study has a purpose of investigating the effect of LEAA (Leucine-enriched essential amino acids mixture) intake on muscle recovery and muscle fatigue of disabled athletes through examining the wheelchair basketball players.

**Methods:** Ten of wheelchair basketball players with spinal cord injury and amputated participated were recruited. All subjects were exposed to a LEAA (12.0 g/day) or placebo treatment in a double-blind, randomized, and cross-over experiment. We measured the variables related to muscular fatigue or inflammatory response before the intense exercise and followed 4 day of recovery.

**Results:** In our results, there was significant decrease of circulating IL-6 level in LEAA treated group compared to placebo
group. However, there was not alternation of TNF-alpha and CK levels. There was no significant difference in relative values of muscle soreness with ANOVA analysis. However, significant improvement in relative values of whole body & back soreness with Effect size analysis with Cohen's d.

**Conclusions:** Based on our results and previous reports, supplementation of LEAA before and after the intense exercise-induced muscular fatigue could be helpful to reduced muscular soreness and inflammatory response in wheelchair basketball players.

**Keywords:** Wheelchair basketball, Muscular fatigue, Inflammatory, LEAA (Leucine-enriched essential amino acids mixture)

**Background and objectives:** In Rwanda, Nyamagabe and Rutsiro districts have stunting prevalence rates above the WHO critical threshold of 40%. The one UN Nutrition projects 'multi-multi' was implemented between August 2014 and December 2016, to effectively reduce the stunting levels in the two districts. Impact of siblings on child stunting in targeted households was studied.

**Methods:** A cross-sectional survey sampled 3700 children aged 6-59 months in the two districts. Data were collected between August and September 2016, and were stratified by number of siblings in household. Stunting in children was determined using the World Health Organization (WHO) growth standard 2006 index (HAZ). Data quality was assessed using ENA plausibility check and statistical analysis was performed with SPSS version 24.

**Results:** Stunting was lower in children enrolled in the project 50.9% vs. 54.9%, P > 0.05. Trend is much significant 46.4% vs. 54.1%, p<0.05 in children enrolled in the project with no sibling under five years in the household. Stunting level was reversed among households having targeted children with older siblings not enrolled in the programme (57.4% vs.56.3%). Stratification by age in household with older siblings, indicates that the older the child is (12-24 months) the more affected he or she is.

**Conclusions:** Household size especially, presence of one or more siblings under five years is a social determinant of child stunting. Additional research is required on quantifying the impact of siblings on child undernutrition for future programme targeting.

**Keywords:** Siblings, Stunting, 'multi-multi'

**Further collaborators:**

Cyprian Ouma;

Alex Kimani;

Allison Oman.

**Background and objectives:** Policy-makers increasingly call for evidence of cost-effectiveness in international aid and nutrition programming. Guidance on tailoring methods to policy relevant questions is limited, making it challenging to determine cost-effectiveness in programmatic settings. As part of the Food Aid Quality Review (FAQR) funded by USAID/Food For Peace Office (FFP), we describe cost-effectiveness research methods applied to a preventive supplementary feeding trial comparing four specialized nutritious food aid products.

**Methods:** A cluster-randomized trial enrolled 6,092 children at six months old. Each enrollee received one of four isocaloric foods (three Fortified Blended Foods and one Ready-to-Use-Supplementary Food) with monthly anthropometric measurements for 18 months. Outcomes relevant to cost-effectiveness analysis include number of child-months and number of children ever diagnosed as wasted (Weight-for-Height Z score<-2) or stunted (Height-for-Age Z score<-2). Using an Activity-Based-Costing-Ingredients (ABC-I) approach, we collected costs associated with program fixed investments, food commodities, transportation, storage, distribution, repackaging, programmatic activities, and beneficiary and volunteer opportunity costs. The Incremental Cost-Effectiveness Ratio (ICER) for each cost-effectiveness measure was calculated across the four arms. Cost-effectiveness measures and their formulas were determined based on the nature of the program and outcomes of interest.

**Results:** Because a child could fall into wasting or stunting more than once over 18 months, we assess the programmatic "cost per child never wasted" and "cost per child never stunted". The ICER formula used to compare across arms is "Difference in total program cost per child / Difference in (number of children ever diagnosed with wasting or stunting / number of children)". Because the length of time during which a child was malnourished is important to long-term growth and development, we will determine "cost per child-month of stunting averted" and "cost per child-month of wasting averted". The ICER formula is "Difference in total program cost per child / Difference in (number of total child-months with stunting or wasting/number of children)".

**Conclusions:** These methods offer rigor to cost-effectiveness research design, including cost data collection and creation of appropriate cost-effectiveness measures. Meaningful cost-effectiveness estimates produced by the research community will in turn

<footnote>(1)Shen, Ye; (2)Cliffer, Ilana; (3)Suri, Devika; (4)Vosti, Stephen; (5)Webb, Patrick; (5)Rogers, Beatrice.</footnote>

<footnote>(1)MSPH. Friedman School of Nutrition Science and Policy. Tufts University. USA.; (2)MPH. Friedman School of Nutrition Science and Policy. Tufts University. USA.; (3)MS. MPH. Friedman School of Nutrition Science and Policy. Tufts University. USA.; (4)PhD. University of California-Davis. USA.; (5)PhD. Friedman School of Nutrition Science and Policy. Tufts University. USA.</footnote>
lead to real impact on better-designed-and-managed nutrition programs and policies in international development.

**Keywords:** cost-effectiveness, supplementary feeding, stunting, wasting, food aid

PLASMA ALANINE AMINOTRANSFERASE (ALT) ACTIVITY IS A RELIABLE BIOMARKER FOR THE RISK OF INSULIN RESISTANCE AND DIABETES PREDISPOSITION

(1)Goda, Toshinao; (2)Honma, Kazue; (3)Tohyama, Kazushige;

(4)Mochizuki, Kazuki.

(1)PhD. Professor. School of Food and Nutritional Sciences. University of Shizuoka. Japan; (2)PhD. Research Assistant Professor. School of Food and Nutritional Sciences. University of Shizuoka. Japan; (3)SBS Shizuoka Health Promotion Center. Japan; (4)PhD. Professor. Department of Local Produce and Food Sciences. University of Yamanashi. Japan.

**Background and objectives:** The onset of diabetes is reportedly related with insulin resistance and uncontrolled gluconeogenesis in the liver. In this study, we examined whether elevated plasma concentration of alanine aminotransferase (ALT), a hepatic key enzyme for gluconeogenesis, is associated with the increases in blood HbA1c levels during the 5-year follow-up period after health checkups in middle-aged apparently healthy Japanese men.

**Methods:** Associations between the baseline clinical parameters or lifestyle factors (categorized in Q1-Q4) and the increase in blood HbA1c levels were determined in 1,729 men aged 35-54 years by logistic regression analyses. We also conducted In vitro analysis where human liver-derived HepG2 cells were cultured with 0.5-100 nmol/L insulin for 8h, and the mRNA levels of ALT gene and the extent of histone acetylation around the gene were measured.

**Results:** Multivariate logistic regression analyses showed that among the clinical parameters and lifestyle factors, the baseline plasma ALT activity was most explicitly associated with the increase in blood HbA1c levels, and the odds ratios for the increment of HbA1c of 0.5% or more in 5 years were 1.71 and 2.64, in the quartile groups Q3 (p = 0.018) and Q4 (p < 0.001) for ALT, respectively, as compared with Q1. In vitro analysis of human liver-derived HepG2 cells showed that the mRNA levels of ALT2, but not ALT1, were reduced by insulin in a dose dependent manner, and that the insulin-induced suppression of ALT2 mRNA levels was in association with decreases in histone H3 and H4 acetylation surrounding the ALT2 gene.

**Conclusions:** These results suggest that plasma ALT activity is a reliable biomarker for the risk of insulin resistance and diabetes predisposition in middle-aged apparently healthy Japanese men.

**Keywords:** ALT, diabetes, insulin resistance, liver

BENEFICIAL EFFECT OF PERSONALISED LIFE-STYLE ADVICE AS COMPARED TO GENERIC ADVICE ON WELLBEING AMONG ACTIVE DUTCH SENIORS - A PILOT STUDY

(1)de Hoogh, Iris; (2)Doets, Esmée; (3)Holthuysen, Nancy; (4)Wopereis, Suzan; (5)Verain, Muriel; (4)Boorsma, André; (6)Bouwman, Emily; (7)Pasman, Wilrike; (4)van Erk, Marjan; (8)Reinders, Machiel.

(1)MSc. Nutrition Scientist. Microbiology & Systems Biology. TNO. The Netherlands.; (2)PhD. Wageningen Food & Biobased Research. Wageningen University & Research. The Netherlands.; (3)BSc. Wageningen Food & Biobased Research. Wageningen University & Research. The Netherlands.; (4)PhD. Microbiology & Systems Biology. TNO. The Netherlands.; (5)Wageningen Economic Research. Wageningen University & Research. The Netherlands.; (6)MSc. Wageningen Economic Research. Wageningen University & Research. The Netherlands.; (7)PhD. Microbiology & Systems Biology. TNO. the Netherlands; (8)PhD. Wageningen Economic Research. Wageningen University & Research. The Netherlands.

**Background and objectives:** Ageing is associated with a gradual loss of skeletal muscle mass and function. It is estimated that sarcopenia affects 30% of seniors over 60 years. Also, 60% of Dutch seniors suffer from overweight or obesity. Both obesity and sarcopenia can be treated with a healthy diet and regular physical exercise.

Previous studies have shown that personalised advice is more effective than general information for improving healthy behavior. Personalisation of lifestyle advice can be done by using individual's health status and behavior (biological factors), and by using personal characteristics (socio-psychological factors). The primary focus of this study was to demonstrate whether personalisation of lifestyle advice as compared to generic advice improves measures of wellbeing in a population of active seniors.

**Methods:** At total of 59 seniors aged 60 years or older and with a self-reported BMI of 20-30 kg/m2 were recruited. The study was designed as a 9-week single-blind randomised controlled trial. At baseline and end, biological, genetic and socio-psychological measurements were performed. Food intake and compliance were measured three times during the study. The intervention group (n=30) received Personalised dietary and lifestyle Advice (PA), based on their dietary intake, phenotype and genotype. The control group (n=29) received Generic Advice (GA) for improving their muscle health.

**Results:** Scores on the Short Physical Performance Battery (SPPB) increased significantly in both the PA and GA group. Decrease in body fat percentage and hip circumference was significantly greater in the PA group. Waist circumference decreased significantly in the PA group, but the difference between the PA and GA group was not significant. There were no differences in changes of self-perceived health between groups. However, in the PA group, scores for motivation and resilience increased significantly.

**Conclusions:** This study showed that both the PA and GA group improved their muscle health as shown by improved scores
on SPPB. However, the PA group showed a stronger beneficial decline in body fat percentage and hip circumference as compared to the GA group. This indicates that personalised lifestyle advice results in additional health benefits in a population of active seniors as compared to general advice.

**Keywords:** personalized advice, elderly, lifestyle, nutrition, muscle health

**Further collaborators:**

Jos van den Puttelaar, MSc. Wageningen Economic Research. Wageningen University & Research. the Netherlands.

Koen Hogenelst, PhD. Microbiology & Systems Biology. TNO. the Netherlands.

Marielle Timmer, MSc. Wageningen Food & Biobased Research. Wageningen University & Research. the Netherlands.

144/1802

NUTRIENTS AND DNA METHYLATION ACROSS THE LIFE COURSE: A SYSTEMATIC REVIEW OF STUDIES IN HUMANS

(1)Braun, Kim; (2)Mandaviya, Pooja; (3)Nano, Jana; (4)Thomas, Carolin;

(5)Troup, Jenna; (6)Muka, Taulant; (7)Bramer, Wichor; (8)Heil, Sandra;

(9)Franco, Oscar; (9)Voortman, Trudy.

(1)RD. MSc. Department of Nutrition. Harvard T.H. Chan School of Public Health. USA.; (2)MSc. Department of Clinical Chemistry. Erasmus University Medical Center. The Netherlands.; (3)MD, MSc. Department of Epidemiology. Erasmus University Medical Center. The Netherlands.; (4)MSc. Department of Epidemiology. Erasmus University Medical Center. The Netherlands.; (5)MSc. Research and Development. Metagenics Inc. USA; (6)MD, PhD. Department of Epidemiology. Erasmus University Medical Center. The Netherlands.; (7)Medical Library. Erasmus University Medical Center. The Netherlands.; (8)PhD. Department of Clinical Chemistry. Erasmus University Medical Center. The Netherlands.; (9)PhD. Department of Epidemiology. Erasmus University Medical Center. The Netherlands.

**Background and objectives:** DNA methylation can be modified by environmental factors, including nutrition. The susceptibility to change of DNA methylation in response to nutrition is particularly high during early life. Nevertheless, nutrition has also been reported to be associated with DNA methylation in other stages of the life course, for instance, during adolescence and adulthood. In order to gain more insight in effects of several nutrients on DNA methylation across the life course, a comprehensive overview of the current knowledge is of importance. Therefore, we conducted a systematic review on the relationship between nutrients and DNA methylation in humans across the life course.

**Methods:** The literature search was designed by an experienced biomedical information specialist. Six bibliographic databases (Embase.com, Medline (Ovid), Web-of-Science, PubMed, Cochrane Central and Google Scholar) were searched. We selected studies that examined the association between nutrients (blood levels; dietary intake; or dietary supplements) and DNA

methylation (global, site specific or genome-wide) in humans of all ages. We did not apply any restrictions on year of publication or language. All abstracts, full text selection and data extraction was performed by two independent reviewers, with a third reviewer available for disagreements.

**Results:** Of the 3774 references identified, 109 studies met our inclusion criteria. Of the included studies, 31 studies investigated the association between maternal nutrition and offspring DNA methylation, nine studies were carried out in subjects during infancy, childhood, or adolescence, and 69 studies were performed in adults. Several candidate gene and epigenome-wide association studies identified effects of nutrients, including folate, fatty acids, and vitamin D, on DNA methylation of CpG sites known to be involved in several health aspects, such as embryonic development and cancer.

**Conclusions:** To date, promising results have been reported in the field of nutrition and DNA methylation in humans at different stages across the life-course. Especially for nutrients known to be involved in one-carbon metabolism, such as folate, but also others, such as fatty acids and vitamin D. However, further large-scale studies of high quality are needed to expand our understanding on the role of nutrition in DNA methylation and its effects on health and disease.

**Keywords:** DNA methylation, systematic review, epidemiology, life-course, epigenetics.

**Conflict of Interest Disclosure:** The authors Kim V.E. Braun, Trudy Voortman, Jenna Troup, Taulant Muka, and Oscar H. Franco work in ErasmusAGE, a center for ageing research across the life course funded by Nestlé Nutrition (Nestec Ltd.), Metagenics Inc. and AXA. The authors report no conflict of interest related to this work.

144/1837

THINK FLEXIBLE TO ADDRESS NUTRITION RESEARCH CHALLENGES

(1)Tanguy, Jérôme; (2)Crabbé, Rafael; (3)Chow, Shein-Chung.

(1)MSc. Biostatistician. Nestle Research Center. Clinical Development Unit. Lausanne. Switzerland; (2)MD. Head of Clinical Development Unit. Nestle Research Center. Clinical Development Unit. Lausanne. Switzerland; (3)PhD. Professor of Biostatistics and Bioinformatics. Department of Biostatistics and Bioinformatics. Duke University School of Medicine. Durham. North Carolina. USA.

**Background and objectives:** Human studies are essential to obtain evidence of beneficial effects of nutritional interventions on health. Nutrition clinical trials have specific challenges to overcome such as population selection, lack of learning phases, small beneficial effects, high heterogeneity of the response and difficulties in finding appropriate endpoints. We believe that this challenging framework favors the need of exploratory but optimized clinical trials: It creates significant opportunities, but also barriers, for the application of adaptive design methods.
**Methods:** This work highlights specificities and challenges in nutritional clinical trials. The application of adaptive design methods is then evaluated in terms of benefits, challenges and obstacles during implementation. Practical recommendations for implementation are also presented.

**Results:** In practice, adaptive design provides the opportunity to modify certain aspects of the trial design whilst the study is ongoing, without violating the quality and the integrity of the data. It allows the investigator to correct wrong assumptions made at the beginning of the trial such as power calculation for sample size; it helps to select the most promising option early with limited number of subjects available at interim; and it provides the opportunity to react earlier to surprise either positive (e.g., strong benefits) or negative (e.g., safety concern or futility).

Compared to traditional designs implementation, adaptive designs require additional efforts at each stage of the trial. It requires careful planning. Simulation-based evaluation of the trial and formal assessment of trial options with quantitative comparison of design choices are recommended. It requires a precise operational execution, with appropriate technology. A close collaboration and coordination is needed among different functional and operational teams during the course of the study. Underlying theoretical complexity of adaptive design implies a solid statistical ground and a careful interpretation of the results. Major adaptations or modifications to a trial could prevent from answering to the scientific questions that the original trial intended to answer.

**Conclusions:** Authors hope this work will allow a better understanding and adoption of the adaptive design methodology by the nutrition scientific community, and will contribute to the design of more efficient and ethical optimized clinical trials.

**Keywords:** Clinical Research, Nutrition specificities, Adaptive Design, Flexible trials

build capacity to identify and measure results of nutrition-related interventions, 2) address knowledge gaps on effectiveness and cost-effectiveness of nutrition interventions, and how they can be delivered at scale, 3) support civil society organizations (CSOs) to strengthen their ability to carry out effective evaluations of their nutrition-related interventions, and 4) disseminate evidence of best practice for impact. After conducting a gap analysis to determine NEEP priorities, PATH (following consultation with DFID) allocated grants and provided technical assistance to local CSOs, tracking performance and compliance throughout the program. Altogether, NEEP supported robust evaluation of 18 nutrition-related interventions in 12 different low- and middle-income countries.

**Methods:** Near the halfway point of the program and approximately one month before CSO contracts were scheduled to end, PATH surveyed grantees to identify any common challenges they faced during the evaluation process and how those challenges were overcome. The authors of this paper reviewed qualitative data from these surveys, monthly and quarterly NEEP grantee reports, and other internal documents and compiled a list of key implementation challenges and recommendations for future nutrition evaluation funding projects.

**Results:** Grantees reported challenges related to tracking and accessing intervention participants for evaluation, obtaining an adequate intervention sample size to achieve statistically significant results, experiencing delays with local internal ethical review board approval, political unrest creating unsafe conditions for evaluators, and transporting anthropometric measurements/equipment. PATH recognized that the timeline for these evaluations could be made more flexible to allow for thorough data collection and publication, and accommodate for environmental challenges beyond the evaluators' control.

**Conclusions:** The NEEP administration and implementation process was challenging, but provided valuable lessons that are transferrable to other nutrition program evaluators and funders. NEEP presented a successful model for an “ongoing external presence” for CSO grantees, ensuring quality and consistency in monitoring and evaluation from the initial planning stages to project completion.

**Keywords:** Evaluations, nutrition-sensitive, CSOs, PATH

**Conflict of Interest Disclosure:** This abstract was produced on the lessons learned through the project (NEEP) supported by the UK aid and the UK Government; however, the views expressed do not necessarily reflect the UK Government's official views or policies.

# THE NEEP APPROACH: CLOSING RESEARCH GAPS AND BUILDING CSO CAPACITY THROUGH EVALUATION

(1)Kothari, Monica; (2)Mackison, Dionne; (3)Houston, April; (4)Fimbi, Sandrine; (5)Perry, Abigail; (6)Musselman, Brianna; (7)Lionetti, Denise.

(1)MS/MPH, Senior Monitoring and Evaluation Adviser, PATH, Washington, DC, USA; (2)PhD. Nutrition Adviser. Nutrition Team. DFID. London. United Kingdom; (3)MS. Communications Consultant, PATH, Washington DC.USA; (4)Finance and Operations Team Lead, PATH, Washington DC.USA; (5)MSc. Senior Nutrition Adviser. Nutrition Team. DFID. London. United Kingdom; (6)MA, Senior Program Assistant, PATH, Washington. DC. USA; (7)MPH, Project Director, PATH, Washington. DC. USA.

**Background and objectives:** The Nutrition Embedding Evaluation Program (NEEP) was a four-year program that began in October 2013, funded by the UK Department for International Development and managed by US-based international health organization PATH. The objectives of NEEP were to: 1)
EFFECT OF A FUNCTIONAL MILK FAT ENRICHED
IN RUMENIC ACID AND VACCENIC ACID ON LI-
PID AND CARBOHYDRATE METABOLISM IN
RATS FED A HIGH-FAT DIET

(1)Bernal, Claudio; (2)Sain, Juliana; (3)Gerstner, Carolina; (4)Lavandera, Jimena; (5)Scanarotti, Ignacio; (6)González, Marcela; (7)Antonacci, Liliana; (8)Gagliostro, Gerardo.

(1)PhD. Biochemist. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. National Scientific and Technical Research Council. Argentina.; (2)PhD. Biotechnologist. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. National Scientific and Technical Research Council. Argentina.; (3)PhD student. Biochemist. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. National Scientific and Technical Research Council. Argentina.
(4)PhD. Biologist. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. National Scientific and Technical Research Council. Argentina.; (5)Biotechnologist student. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. Argentina.; (6)PhD. Biochemist. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. Argentina.; (7)Msc. Agronomist. National Institute of Agricultural Technology. Argentina.; (8)PhD. Agronomist. National Institute of Agricultural Technology. Argentina.

**Background and objectives:** Intake of high-fat (HF) diets has been associated with steatosis, hepatic lipid alterations and insulin resistance in humans and in experimental animal models. Since rumenic acid (RA) and vaccenic acid (VA), naturally present on dairy products, showed to induce health benefits on lipid and carbohydrate metabolism, some efforts have been made to increase their levels in the milk fat (MF) and obtain a functional milk fat (FMF). The aim of the present study is to investigate parameters related to glucose and triacylglycerol (TAG) metabolism in rats fed high-fat diets containing a FMF obtained by PUFA supplementation to dairy cow diets.

**Methods:** Male Wistar rats were fed (60-d) with S7 (soybean oil, 7%), S30 (soybean oil, 30%), MF30 (soybean oil, 3% + MF, 27%) or FMF30 (soybean oil, 3% + FMF, 27%) diets. Biochemical parameters on serum; TAG levels in liver and skeletal muscle; lipogenic and β-oxidative enzymes activities and expressions in liver; and hepatic TAG secretion rate in vivo were determined. In addition, an oral glucose tolerance test, the metabolites from glycolysis and some key enzyme activities in skeletal muscle were assessed. Statistical differences (p<0.05) were tested by ANOVA.

**Results:** Compared to S7, S30 diet induce TAG accretion in liver (+36%) and skeletal muscle (+61%), decreased muscle glu-

cose utilization by a lower flux through hexokinase (-53%) and 6-phosphofructokinase (-49%), glucose intolerance and hyperinsulinemia (+48%). The FMF30 group showed lower levels of TAG in liver (-26%) associated with an enhanced β-oxidation (CPT-I gene expression +122%) and a higher TAG secretion rate (+31%), compared to S30. In muscle, FMF30 also reduced the TAG levels (-49%) and increased the flux through the 6-phosphofructokinase (+116%) improving the glucose utilization.

**Conclusions:** In conclusion, the FMF prevented the TAG accretion induced by high fat diet consumption and improve the glucose utilization in skeletal muscle.

**Keywords:** rumenic acid, vaccenic acid, functional milk fat, lipid metabolism, glucose metabolism

MATERNAL CONJUGATED LINOLEIC ACID MOD-
ULATES TRIACYLGLYCERIDE METABOLISM IN
ADULT OFFSPRING RATS

(1)Bernal, Claudio; (2)Lavandera, Jimena; (3)Gerstner, Carolina;

(4)Sain, Juliana; (5)Fariña, Ana; (5)González, Marcela; (4)Vera Candioti, Luciana; (6)Mancini, Sonia; (7)Williner, Maria.

(1)PhD. Biochemist. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. National Scientific and Technical Research Council. Argentina.; (2)PhD. Biologist. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. National Scientific and Technical Research Council. Argentina. (3)PhD student. Biochemist. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. National Scientific and Technical Research Council. Argentina. (4)PhD. Biotechnologist. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. National Scientific and Technical Research Council. Argentina. (5)PhD. Biochemist. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. Argentina. (6)PhD student. Biotechnologist. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. Argentina.; (7)Msc. Biochemist. Food Science and Nutrition. Department of Biological Sciences. School of Biochemistry and Biological Sciences. National University of Litoral. Argentina.

**Background and objectives:** Conjugated linoleic acid (CLA) might regulate the lipid liver and adipose tissue accretion. Since there is an association between maternal nutrition, fat depots and risk of offspring chronic disease, the aim was investigate the effect of maternal CLA consumption on triacylglycerol (TAG) regulation and some inflammatory parameters in adult male rat offspring receiving or not CLA.
**Methods:** Female Wistar rats were fed control (C) or CLA-supplemented (1%,w/w) diets during 4 weeks before and throughout pregnancy and lactation. After weaning, male offspring of CLA rats were fed C or CLA diets (CLA/C and CLA/CLA groups, respectively), while C male offspring rats were fed C diet (C/C group) for 9 weeks. Serum TAG concentration, liver TAG content, lipogenic and oxidative enzyme activities, gene expressions, transcriptional factors (SREBP1c, PPARα and PPARγ) and inflammatory cytokines mRNA levels; and epididymal white adipose tissue (EWAT) LPL activity were measured.

**Results:** Serum TAG levels were increased in CLA/CLA (+65%) and CLA/C (+92%) groups, associated with a reduction of EWAT LPL activity (-41%) and (-67%), respectively. Liver TAG levels were decreased in CLA/CLA (-22%) group related to a significant reduction of FAS (-37%) and ACC (-27%) enzyme activities, as well as of FAS (-44%), ACC (-48%), SCD1 (-54%) and SREBP1c (-25%) mRNA levels, without changes in β-oxidation parameters. Even though in CLA/C rats a normal liver TAG levels were founded, FAS (-38%) and ACC (-31%) activities, as well as ACC (-62%) expression were significantly diminished, associated with a lower expression of the key enzyme of β-oxidation (CPT-Ia) (-23%), PPAR-α (-22%) and PPAR-γ (-45%). Liver IL-1β gene expression showed a significant decrease in both, CLA/CLA and CLA/C groups. No changes were observed in IL-6 mRNA levels.

**Conclusions:** These results demonstrate that maternal CLA supplementation programs male rat offspring liver lipid metabolism leading to a prevention of the TAG accretion in adipose tissue and liver. The present findings could be important to develop some dietary strategies to reduce the incidence of obesity and fatty acid liver disease in humans.

**Keywords:** Conjugated linoleic acid, Offspring, Triacylglycerol, Fetal programming, Lipid metabolism

144/1879

## SYSTEMATIC REVIEW ON THE MARKERS OF GLYCAEMIC EXPOSURE IN THE NON-DIABETIC POPULATION

(1)Feskens, Edith; (2)Brennan, Lorraine; (3)Dussort, Pierre; (4)Mela, David J.; (5)Rabbani, Naila; (6)Rathmann, Wolfgang; (7)Respondek, Frédérique; (8)Theis, Stephan; (5)Thornalley, Paul; (9)Vinoy, Sophie.

(1)Division of Human Nutrition and Health. Wageningen University. The Netherlands; (2)PhD. Professor. University College Dublin. Dublin. Ireland.; (3)PhD. Coordinator. ILSI Europe. Brussels. Belgium.; (4)PhD.Scientist.Unilever. Vlaardingen. The Netherlands.; (5)PhD. Professor. Warwick University.Coventry. United Kingdom.; (6)MD. Vice-Director. German Diabetes Center. Dusseldorf. Germany.; (7)PhD. Scientist. Tereos. Thumeries. France; (8)PhD. Scientist. Südzucker Group. Mannheim. Germany.; (9)PhD. Scientist. Mondelēz International. Saclay. France.

**Background and objectives:** Reducing dietary glycaemic exposures in the general population is increasingly supported as a preventive approach to address the growing global burden of Type

2 diabetes. To study the impact of dietary modifications affecting glycaemic exposures, the availability of established biomarkers suitable for use in non-diabetic populations is essential. Therefore, our objective was to review and characterize alternative markers of sustained glycaemic exposures that could be used to evaluate and compare the glycaemic impact of dietary interventions among non-diabetic populations.

**Methods:** The protocol has been described in the PROSPERO database of systematic reviews, register number CRD42017037408. The following markers/measurements were pre-selected: continuous glucose monitoring, di-carbonyl stress, 1,5-anhydroglucitol, fructosamine, glycated albumin/proteins, advanced glycated end products (AGEs), HbA1c, and metabolomics. Systematic searches were done for publications of human studies in MEDLINE and EMBASE published until July 2016, to identify research where these markers had been measured in relation to diet as part of intervention or observational studies. Results were interpreted with respect to sensitivity, time-responsiveness, conditions and range of response.

**Results:** The systematic search generated 6547 unique records. After the screening process, 111 papers were included, ranging from only 4 for 1,5-anhydrioglucitol to 30 for AGEs. The selected papers highlighted the paucity of available data for these markers as tools to assess the glycaemic exposure in non-diabetic populations.

**Conclusions:** Considerable additional work is needed to validate the use of any of these as biomarkers for assessment of glycaemic exposure in non-diabetic populations. Such work includes demonstration of dose response, sensitivity and specificity, development of quantitative measurement assays and measurement in larger cohorts.

**Keywords:** biomarkers, glycemic exposure, type 2 diabetes, prevention, carbohydrates

**Conflict of Interest Disclosure:** This work was conducted by an expert group of the European branch of the International Life Sciences Institute, ILSI Europe. This publication was coordinated by the Dietary Carbohydrates Task Force. Authors DM, FR, ST and SV are employed by commercial organizations that manufacture carbohydrate-containing foods and beverages or ingredients used in making these. The expert group received funding from the ILSI Europe the Dietary Carbohydrates Task Force members. Industry members of this task force are listed on the ILSI Europe website at http://ilsi.eu/task-forces/nutrition/dietary-carbohydrates/. Experts are not paid for the time spent on this work; however, the non-industry members within the expert group were offered support for travel and accommodation costs from the Dietary Carbohydrates Task Force to attend a meeting to discuss the manuscript and a small compensatory sum (honoraria) with the option to decline. The research reported is the result of a scientific evaluation in line with ILSI Europe's framework to provide a precompetitive setting for public-private partnership (PPP). ILSI Europe facilitated scientific meetings and coordinated the overall project management and administrative tasks relating to the completion of this work. The opinions expressed herein and the conclusions of this publication are those of the authors and do not necessarily represent the views of ILSI Europe nor those of its member companies.

**Further collaborators:**

Coen Stehouwer.

cda.stehouwer@maastrichtuniversity.nl.

MD. Professor. Department of Internal Medicine. Maastricht University, Maastricht. The Netherlands.
# GHRELIN AND LEPTIN SERUM LEVELS AND ANTHROPOMETRIC NUTRITIONAL STATUS IN RELATION TO HELICOBACTER PYLORI GENOTYPE AND GASTRIC PATHOLOGY OF DYSPEPTIC ADULTS

(1)Mantero, Paula; (2)Marchesi Olid, Liliana; (3)Matus, Gonzalo; (3)Corti, Rodolfo; (4)Cabanne, Ana; (5)Zerbetto de Palma, Gerardo; (6)Piskorz, María Marta; (7)Zubillaga, Marcela; (8)Janjetic, Mariana; (7)Goldman, Cinthia.

(1)Biochemist. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Cátedra de Física. Argentina; (2)Nutritionist. Universidad de Buenos Aires. Facultad de Medicina. Escuela de Nutrición. Argentina.; (3)MD. Hospital de Gastroenterología "Dr. Carlos Honorino Udaondo". Esophagus-Stomach Section. Argentina.; (4)MD. Hospital de Gastroenterología "Dr. Carlos Honorino Udaondo". Pathology Unit. Argentina.; (5)Biochemist. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Cátedra de Física. Argentina. Universidad de Buenos Aires. CONICET. Instituto de Microbiología y Parasitología Médica (IMPAM). Argentina.; (6)MD. Hospital de Clínicas "José de San Martín". Gastroenterology Unit. Argentina.; (7)PhD. Biochemist. Pharmacist. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Cátedra de Física. Argentine National Scientific and Technological Research Council (CONICET). Argentina.; (8)PhD. Nutritionist. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Cátedra de Física. Facultad de Medicina. Escuela de Nutrición. Argentine National Scientific and Technological Research Council (CONICET). Argentina.

**Background and objectives:** Hormonal regulation of appetite could be affected by the bacterium Helicobacter pylori. The objective of this study was to evaluate the association of H. pylori genotype and the type of gastric pathology, with serum ghrelin and leptin concentrations and Body Mass Index (BMI).

**Methods:** Fasted dyspeptic adults referred for an upper gastrointestinal endoscopy were enrolled. Height, weight and nutrient intake were assessed. Serum total ghrelin and leptin levels were analyzed by enzyme-linked immunosorbent assay. 13C-Urea Breath Test was performed and four gastric biopsies were obtained during endoscopy for histopathology and H. pylori DNA amplification and genotyping. Data analysis was performed using the χ2, Mann-Whitney U, and Kruskal-Wallis tests, Spearman's correlation and linear regression.

**Results:** 163 patients (40.8 ± 14.0y), 98/65 females/males, were included. H. pylori prevalence was 53.4% (CI95%; 45.7 - 65.8%). Neither nutrient intake nor BMI differed significantly between groups. Serum ghrelin concentrations were significantly lower in H. pylori+ patients [median 311.0 pg/mL (IQR; 230.0 - 385.5)], independently of their cagA genotype, than in H. pylori- [median 355.0 pg/mL (IQR; 253.8 - 547.8)] (P = 0.025), even after adjusting for BMI and gender (P = 0.03). Leptin levels did not differ significantly between infected and uninfected patients [median 1.84 (0.80 - 4.85) vs 1.84 (0.50 - 5.09), (P = 0.87)]. The type and severity of gastric pathology in the corpus was associated with lower

ghrelin serum levels (P = 0.04); whereas, gastric pathology of the antrum was not associated to ghrelin levels (P = 0.08).

**Conclusions:** H. pylori infection, independently of its cagA genotype, and the severity of gastric pathology of the corpus, are associated with lower ghrelin serum concentrations in dyspeptic patients, emphasizing its role on appetite regulation.

**Keywords:** H. pylori, cagA genotype, ghrelin, leptin, gastric pathology

# EXPLORING VARIATIONS IN HEMOGLOBIN CONCENTRATION AND MEASUREMENT – THE HEMOGLOBIN MEASUREMENT (HEME) WORKING GROUP

(1)Moorthy, Denish; (2)Namaste, Sorrel; (3)Rappaport, Aviva.

(1)Technical Advisor. Micronutrients. Spring Project. USA.;

(2)Team Lead. Technical Advisor. Anemia. Spring Project. USA.;

(3)Consultant. Spring Project. University Of British Columbia. Canada.

**Background and objectives:** Measurement of hemoglobin concentration is common practice for monitoring and evaluating anemia reduction programs. Accuracy of hemoglobin measurement may vary widely. In order to improve the accuracy and quality of hemoglobin measurement across contexts, the USAID-funded Strengthening Partnerships, Results: **Methods:** Data were identified from cross-sectional studies among different population groups (children 6 -59 months, and pregnant and non-pregnant women 18-45 years) to assess precision, agreement, and measures of clinical validity to diagnose anemia.

**Results:** We identified data from nine studies conducted in Asia (Lao PDR, Cambodia, and India), Africa (Ghana, Rwanda, and Republic of The Gambia), and the Americas (Mexico, Guatemala, and USA) that compared hemoglobin measurements done using multiple devices (hematology autoanalyzers, and HemoCue devices) in different settings (laboratory and field). Priority research topics generated include: (1) factors influencing hemoglobin concentration, specifically capillary versus venous blood samples, type of analyzer, different models of the Hemocue devices, collection methods, and effect of sex, age, and climate.

**Conclusions:** This multi-investigator, multi-country research collaboration is the first of its kind to study variations in hemoglobin measurement. The findings from the study will contribute to the World Health Organization (WHO)'s review of its global guidelines for hemoglobin thresholds used to define anaemia at the individual and population level.

**Keywords:** Hemoglobin Anemia Measurement Variation Collaboration

**Further collaborators:** Donnie Whitehead. PhD. Centers for Disease Control and Prevention. USA. Zuguo Mei. MD. Centers for Disease Control and Prevention. USA. Sonja Hess. PhD. University of California at Davis. USA. Crystal Karakochuk. PhD. University of British Columbia. Canada. Sam Newton. MD. Kwame Nkrumah
University of Science and Technology. Ghana. Elizabeth Abu-Haydar. MPH. PATH. USA. Ignacio Mendez Gomez-Humaran. MSc. National Public Health Institute. Mexico. Dora Ines Mazariegos Cordero. MSc. Instituto de Nutrición de Centro América y Panamá. Guatemala. Megan Parker, PhD. PATH. USA. Rita Wegmüller. PhD. Medical Research Council. The Gambia. Lynnette Neufeld. PhD. Global Alliance for Improved Nutrition. Geneva. Teemar Fisseha. MPH. USAID-funded SPRING Project. John Snow Inc. USA.

## HARMONIZATION OF DATA COLLECTION ACROSS 4 COUNTRIES - THE ENRICH EXPERIENCE

(1)Owais, Aatekah; (2)Begum, Khadija; (3)Kekre, Priya; (4)Sellen, Daniel.

(1)MPH PhD. Project Statistician. Department of Anthropology. University of Toronto. Canada; (2)MBBS MPH PhD. Research Coordinator. Department of Anthropology. University of Toronto. Canada; (3)MPH. Project Technical Assistant. Department of Anthropology. University of Toronto. Canada; (4)PhD. Director. Centre for Child Nutrition and Health. University of Toronto. Canada.

**Background and objectives:** Undernutrition remains a major contributor to maternal and childhood morbidity and mortality. Enhancing Nutrition Services to Improve Maternal and Child Health in Africa and Asia (ENRICH) is a multi-country, broad-spectrum and integrated intervention program designed to help member countries achieve their global nutrition targets.

**Methods:** As part of ENRICH impact evaluation, a baseline assessment was conducted via household surveys (HHS) using a quasi-experimental design. In three of the four countries (Bangladesh, Kenya, Tanzania), a comparison area likely to have baseline characteristics similar to the intervention area was selected. In Pakistan, no comparison area was planned due to contextual reasons.

**Results:** The survey questionnaires were developed by University of Toronto (UofT) in coordination with World Vision Canada (WVC), ensuring consistency in tools, variables and response sets across settings. In-country research partners (CRPs) in each of the four countries carried out local contextualization and translation of the questionnaires. Data collection at all sites was conducted via Open Data Kit (ODK) platform on android tablets. English and predominant local language ODK templates for field-based entry of HHS were designed in Excel, converted into XML format, and uploaded to country-specific secure ODK Aggregate servers. Blank ODK forms were then accessed by field teams on their android tablets to be filled in during data collection in each country. Completed forms were saved on the tablets until uploaded to the ODK Aggregate server at the end of each working day. Multiple challenges were encountered and met across settings.

**Conclusions:** ODK is an effective and efficient platform for collecting large amounts of data across different geographical sites in a relatively short period of time under a range of field challenges. Data thus collected were of high quality, with minimal effort required for data cleaning and management. Using ODK also allowed for rapid, real-time updates to data collection tools based on feedback from enumerator training and first few days of data collection in each country. Strong training, supervision and real-time international coordination was key to maximizing the potential and flexibility of ODK platforms for complex and large scale evaluations in low-income settings.

**Keywords:** harmonization of data collection, maternal and child nutrition, nutrition intervention program, ODK, multi-country

## PARATHYROID HORMONE AND FETAL LENGTH IN A PREGNANCY COHORT IN DHAKA, BANGLADESH

(1)Qamar, Huma; (2)Perumal, Nandita; (3)Papp, Eszter; (4)Gernand, Alison D; (5)Mahmud, Abdullah Al; (6)Roth, Daniel.

(1)BSc. Graduate Student. Department of Nutritional Sciences. University of Toronto. Canada.; (2)MPH. Graduate Student. Dalla Lana School of Public Health. University of Toronto. Canada.; (3)PhD. Research Manager. Centre for Global Child Health. Hospital for Sick Children. Canada.; (4)PhD, MPH. Nutritional Scientist. Department of Nutritional Sciences. Pennsylvania State University. USA.; (5)MSc. Scientist. Centre for Nutrition and Clinical Services Division. International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b). Bangladesh.; (6)MD, PhD. Clinician Scientist. Centre for Global Child Health. Hospital for Sick Children. Canada.

**Background and objectives:** The prevalence of fetal growth restriction remains high throughout South Asia, but the causes remain unclear. An important measure of fetal skeletal growth is birth length, which has been associated with psycho-motor development and postnatal length gain. Maternal parathyroid hormone (PTH) is a potential modifiable factor that may be a marker of fetal skeletal growth. PTH is a peptide hormone important in regulating calcium homeostasis, mineral metabolism, and bone turnover. In the present study, we aimed to investigate the association of maternal parathyroid hormone (PTH) with infant length at birth in a pregnancy cohort in Dhaka, Bangladesh.

**Methods:** This was a cross-sectional study using mother-infant data and biospecimens obtained at birth from a subset of participants (n=519) enrolled in the Maternal Vitamin D for Infant Growth (MDIG) trial, a dose-ranging prenatal vitamin D supplementation trial in Dhaka, Bangladesh. Infant anthropometry was collected within 48 hours of birth. Laboratory analyses were performed to estimate concentrations of maternal serum 25-hydroxyvitamin D and maternal plasma PTH, magnesium, FGF23, and CRP at delivery. We derived relative maternal calcium intake using a limited food frequency questionnaire. The primary outcome was newborn length-for-gestational-age z-scores (LAZ). Secondary outcomes included other newborn anthropometric indices. Statistical analyses were performed using multivariable linear regression models.

**Results:** At delivery, median maternal perinatal PTH concentration was 3.20 pmol/L (IQR: 1.94-4.80). Mean infant LAZ
at birth was -0.89 (SD=0.99). The associations between maternal PTH concentrations at delivery and infant LAZ as well as other anthropometric indices will be presented.

**Conclusions:** The association of perinatal PTH concentrations and infant anthropometric indices at birth will be summarized. The significance of the findings, including the potential role of PTH as a marker of fetal growth restriction and maternal nutrition status will be discussed.

**Keywords:** Parathyroid hormone, pregnancy, fetal growth restriction, skeletal growth

144/1930

METHODS FOR RIGOROUS IN-HOME OBSERVATION CONDUCTED DURING A FOOD AID COST-EFFECTIVENESS TRIAL IN BURKINA FASO

(1)Cliffer, Ilana; (2)Nikiema, Laetitia; (3)Garanet, Franck; (4)Suri, Devika;

(1)Langlois, Breanne; (5)Shen, Ye; (6)Walton, Shelley; (7)Webb, Patrick;

(7)Rogers, Beatrice.

(1)MPH. Friedman School of Nutrition Science and Policy. Tufts University. USA.; (2)MD. Nutrition Department. Institut de Recherche en Science de la Sante. Burkina Faso.; (3)MD.Nutrition Department. Institut de Recherche en Science de la Sante. Burkina Faso.; (4)MS. MPH. Friedman School of Nutrition Science and Policy. Tufts University. USA.; (5)MSPH. Friedman School of Nutrition Science and Policy. Tufts University. USA.; (6)MPH. RD. Friedman School of Nutrition Science and Policy. Tufts University. USA.; (7)PhD. Friedman School of Nutrition Science and Policy. Tufts University. USA.

**Background and objectives:** To satisfy the need for direct assessments of feeding behaviors, in-home observation methods are gaining in use in global health and nutrition research. Bolstering existing methodological literature, we describe an approach to in-home observation aimed at understanding food preparation and feeding practices during a study comparing the cost-effectiveness of food aid in Burkina Faso.

**Methods:** From November 2014 to October 2016, we conducted four consecutive days of non-participant observations in randomly selected households of project beneficiary children receiving one of four foods in a supplementary feeding programme. Female observers surveyed beneficiary children aged 6-23 months, using a pre-coded grid in 30-minute time increments from 06:00 to 18:00 to record feeding and hygiene practices linked with product use. Qualitative notes were taken on events related to childcare, health and feeding. Post-observation, observers collected self-reported data on feeding practices by administering structured in-depth surveys to each household. The protocol was designed to minimize any Hawthorne effect. Village leaders were informed of observations, and consent was obtained from households. Family members were instructed to ignore the presence of the observer. Observers first spent a day in each household without taking notes to habituate their presence, and lodged elsewhere to reduce influence on normal practices.

**Results:** A total of 209 of the 224 selected households were observed; one household refused observation. Challenges included ensuring that observer presence did not noticeably influence practices, place undue burden on families, or induce social desirability bias. Additionally, it was challenging to create instruments that allowed efficient collection, entry and analysis of data. Advantages to conducting multi-day consecutive observations were habituation of the observed household to the presence of the observer, thoroughness and objectiveness of data, accurate time tracking of beneficiary opportunity costs, and validating reported versus observed behaviors. Observers report that acclimatization after multiple days may reduce social desirability bias, and that village leader buy-in was essential.

**Conclusions:** Multi-day in-home observations can provide rigorous data on observed food preparation and feeding practices and reduce the Hawthorne effect. Techniques, including using tablets, which may streamline data collection, should be further refined to address noted challenges.

**Keywords:** Food Aid, Observation, Supplementary Feeding, Stunting, Wasting

144/1956

CLASSIFICATION OF FOODS INTO FOOD GROUPS IN THE DIETSYS DATABASE

(1)Rodrigues, Marcela Perdomo; (2)Fung, Teresa T; (3)Beato, Gariel c;

(4)Rossato, Sinara L.

(1)Post-graduation program in cardiology and cardiovascular disease. Federal University of Rio Grande do Sul. Porto Alegre. Rio Grande do Sul. Brazil; (2)Department of Nutrition. Harvard T.H. Chan School of Public Health. Boston. Massachusetts. Department of Nutrition. Simmons College. Boston. Massachusetts. USA; (3)Institute of Bioscience. Universidade Estadual Paulista "Julio Mesquita Filho". Botucatu (UNESP). Sao Paulo. Brazil; (4)Presenting author; Department of Nutrition. Harvard T.H. Chan School of Public Health. Boston. Massachusetts. USA. Department of Nursing. Universidade Estadual Paulista "Julio Mesquita Filho". Botucatu (UNESP). Sao Paulo. Brazil.

**Background and objectives:** The DietSys is a diet-data processing system for research that incorporates an international food database to maximize and improve the accuracy of diet-data assessment across countries. However, while using an international database it is difficult to locate a particular food due to the large number of items. This difficulty can be mitigated by classifying food items into food groups. This study aimed to describe the classification of foods into food groups in the DietSys database.

**Methods:** Three food composition tables were merged: the Argentinean food composition table (Argenfoods), the Brazilian food composition table (TACO), and the U.S. Department of Agriculture - Agricultural Research Service (USDA). Food items were classified into 21 food groups: Alcoholic beverages; Non-alcoholic beverages; Cereal and derivatives; Condiments and sauces;
Egg products; Fats and oils; Finfish and shellfish products; Fruits and derivatives; Infant formula & baby food; Legumes and derivatives; Meats and derivatives; Milk and Dairy; Miscellaneous; Mixed dishes; Nuts and seed products; Other; Sugars; Sausages and luncheon meats; Snacks and sweets; Soy products; Vegetables, leafy greens and derivatives.

**Results:** The DietSys database comprised 9851 food items; 4.7% (465 items) from Argenfoods, 6.1% (597 items) from TACO, and 89.2% (8789) from USDA. Food items from Argenfoods fell within 15 of the 21 food groups, with 21.7% categorized as finfish and shellfish products and 20.6% as milk and dairy. From TACO, 26% of food items belonged to miscellaneous, 19% to fruits and derivatives, and 4.6% to meats and derivatives. Of the 9851 food items in the DietSys database, the USDA accounted for 91% of meats and derivatives (N=2060), 96.8% of mixed dishes (1194 items) and 95.9% of snacks and sweets (N=1205 items).

**Conclusions:** The DietSys incorporates a comprehensive database making it feasible to assess diet-data from Argentina, Brazil, and the United States. Classifying foods into food groups expedited and facilitated the search for food items in the large database of the Dietsys system.

**Keywords:** DietSys, DietSys database, food groups database, food groups

insulin signaling cascade were tested using Oil red, immunofluorescence and Western blot, respectively.

**Results:** Both mixes induced apoptosis at 400 µM concentration as compared with lower concentrations. Mix 1 induced macrovesicular steatosis and Mix 2 treatment induced microvesicular steatosis. Higher expression of IL-6, IL-17 and IL-8 and lower expression of Akt protein found in Mix 1 as compared with Mix 2.

**Conclusions:** The results suggest that the mixture of fatty acids associated with the consumption of Mediterranean diet induced less inflammation, apoptosis, insulin resistance and macrovesicular steatosis than a mixture with higher content of SFA in hepatic cells.

**Keywords:** Fatty acids, hepatic cells, inflammation, steatosis.

144/2008

EFFECTS OF THE STIMULATION WITH DIFFERENT MIXES OF MONO, SATURATED AND POLYUNSATURATED FATTY ACIDS ON HEPATIC CELLS

(1)Castellanos Tapia, Lyssia; (2)Gómez Quiroz, Luis Enrique; (3)Aparicio Bautista, Diana Ivette; (3)Tejero, Maria Elizabeth.

(1)Instituto Nacional de Medicina Genómica; (2)PhD. Professor. Universidad Autónoma Metropolitana Iztapalapa. Mexico City. México.; (3)PhD. Researcher. Laboratorio de Nutrigenómica Instituto Nacional de Medicina Genómica. Mexico City. México.

**Background and objectives:** The rate of hepatic disease has increased worldwide over a decade. One of the causes may be the high intake of saturated fatty acids (SFA). These fatty acids acid have been considered as lipotoxic for hepatic cells, induce insulin resistance, inflammation and endoplasmic reticulum stress. Furthermore, oleic and omega 3 polyunsaturated fatty acids may revert the effects of SFA. However, the effect of mixtures of fatty acids that resemble their proportion and concentration in human plasma has not been tested. The purpose of this study is to evaluate the effect of the exposure to two mixtures of fatty acids similar to those found in plasma of subjects exposed to a diet rich in SFA and to a Mediterranean diet, on cultured HepG2 cells.

**Methods:** Cultured HepG2 cells were incubated with Mix 1 (67% saturated, 26% monounsaturated, and 6.5% polyunsaturated) or Mix 2 (55% saturated, 28% monounsaturated and 14.5% polyunsaturated), for 24 h and control. Three different concentrations were tested (100, 200 and 400 µM). Triglyceride accumulation, markers for apoptosis, inflammation and proteins of the

144/2013

NUTRIENT INTAKE, APPETITE SENSATION AND GASTRIC PATHOLOGY IN HELICOBACTER PYLORI INFECTED PATIENTS BEFORE AND AFTER ERRADICATION

(1)Mantero, Paula; (2)Marchesi Olid, Liliana; (3)Giacomantone, Candela; (3)Corti, Rodolfo; (4)Cabanne, Ana; (5)Zubillaga, Marcela; (6)Janjetic, Mariana; (5)Goldman, Cinthia.

(1)Biochemist. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Cátedra de Física. Argentina; (2)Nutritionist. Universidad de Buenos Aires. Facultad de Medicina. Escuela de Nutrición. Argentina.; (3)MD. Hospital de Gastroenterología "Dr. Carlos Bonorino Udaondo". Esophagus-Stomach Section. Argentina.; (4)MD. Hospital de Gastroenterología "Dr. Carlos Bonorino Udaondo". Pathology Unit. Argentina.; (5)PhD. Biochemist. Pharmacist. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Cátedra de Física. Argentine National Scientific and Technological Research Council (CONICET). Argentina.; (6)PhD. Nutritionist. Universidad de Buenos Aires. Facultad de Medicina. Escuela de Nutrición. Facultad de Farmacia y Bioquímica. Cátedra de Física. Argentine National Scientific and Technological Research Council (CONICET). Argentina.

**Background and objectives:** H. pylori could be involved in apetite regulation. Our study aimed to evaluate the nutritional anthropometric status, the appetite sensation and nutrient intake and the gastric pathology in patients before and after H. pylori eradication.

**Methods:** Dispeptic adults referred for an upper gastrointestinal endoscopy were enrolled. We assessed weight and height for body mass index determination. Nutrient intake was evaluated by 24-hour recalls and appetite sensation by a validated questionnaire (SNAQ). We obtained biopsies for gastric pathology analysis. Infected patients returned for control 12 weeks after eradication therapy. During the control we evaluated the same variables as in the first appointment. We used the proportion test and the Wilcoxon Signed Rank Test to perform statistical analysis.

**Results:** 117 patients (43.2±12.8y), 66.7% females (CI95%; 57.7-74.5), were included in this study. H. pylori infection prevalence