## BIOCHEMICAL PARAMETERS OF DISLIPIDEMIC RATS CONSUMING MACAÍBA (ACROCOMIA INTUMESCENS DRUDE)

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

144/1422

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

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

144/1428

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

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

Universidad Autónoma del Estado de México.

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

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

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

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

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

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

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

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

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

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

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

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

144/1430

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

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

Universidad Autónoma del Estado de México.

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

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

144/1460

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

(1)Haskell-Ramsay, Crystal; (1)Jackson, Philippa; (2)Forster, Joanne;

(3)Robertson, Bernadette; (1)Kennedy, David.

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

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

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

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

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

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

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

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

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

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

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

(1)Department of Food and Human Health Sciences. Graduate School of Human Life Science. Osaka City University.; (2)M.Sc. Graduate student. Department of Food and Human Health Sciences. Graduate School of Human Life Science. Osaka City University.; (3)B.Sc. Student. Department of Food and Human Health Sciences. Graduate School of Human Life Science. Osaka City University.; (4)Ph.D. Researcher. Life Science Institute Co. Ltd.; (5)Ph.D. President. Life Science Institute Co. Ltd.; (6)Ph.D. Associate professor. Department of Food and Human Health Sciences. Graduate School of Human Life Science. Osaka City University.

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

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

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

**Conclusions:** These results suggest that DNSM treatment represents a novel tool for the prevention of alcohol-induced liver injury.
**Keywords:** DNA-rich nucleic acid prepared from salmon milt (DNSM), plasma aminotransferases (AST and ALT), liver fibrosis, CYP2E1 activity, alcohol-induced liver injury.

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

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

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

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

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

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

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

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

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

Within-Subject-Factor: time

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

Between-Subject-Factors: RSV/placebo.

Covariates: age, sex, education.

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

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

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

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

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

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

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

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

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

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

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

144/1694

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

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

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

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

Abstracts Presented as Posters

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

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

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

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

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

144/1697

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

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

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

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

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

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

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

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

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

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

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

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

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

144/1715

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

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

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

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

144/1734

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

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

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

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

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

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

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

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

**Further collaborators:**

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

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

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

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

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

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

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

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

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

## 144/1900

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

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

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

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

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

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

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

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

**Further collaborators:**

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

## 144/1905

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

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

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

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

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

**Results:** For cupuaçu pulp the results were: pH of 3.26; Acidity of 0.182 g / 100g; Content of phenolic compounds obtained was 130.80 mg equivalent of gallic acid / 100 g; Flavonoid content was 14.06 mg / 100g, with antioxidant activity 96.50%. Noni: pH 3.95; Acidity of 0.119 g / 100g; Phenolic compounds was 43.30
mg equivalent of gallic acid/100 g; 9.38 mg / 100g of flavonoids;
97.14% antioxidant activity. For cajá-manga: pH of 3.44; Total
acidity 0.068 g / 100g; Content of phenolic compounds obtained
was 160 mg equivalent of gallic acid / 100 g; Flavonoids content
was 2.35 mg / 100 g, with antioxidant activity 95.23%. For ca-
já-manga pulp presents a higher content of phenolic compounds,
but the cupuaçu pulp had a higher content of flavonoids.

**Conclusions:** Fruit pulps were shown to have high levels of phenolic compounds, such as flavonoids and high potential for antioxidant activity.

**Keywords:** Cupuaçu, Noni, Cajá-manga, Lyophilization, Antioxidant.

144/1922

EXPLORING THE FUNCTIONAL PROPERTIES OF
ELEPHANT FOOT YAM (AMORPHOPHALLUS
PAEONIIFOLIOUS) TO AMELIORATE PERIMENO-
PAUSAL SYMPTOMS

(1)Dave, Dr. Mital; (2)Nair, Dr. Sirimavo.

(1)Ph.D. Nutritionist. Department of Foods and Nutrition. The
Maharaja Sayajirao University of Barod. Vadodara. Gujarat. India.;

(2)Ph.D. Associate Professor. Department of Foods and Nutrition.
The Maharaja Sayajirao University of Baroda. Gujarat, India.

**Background and objectives:** Background: Phytoestrogens are natural compounds present in the food, with structural similarity to human estrogen. These compounds have been considered to compete effectively with endogenous estrogens which bind to the ER and stimulate estrogen like effects. They help in ameliorating the perimenopausal and menopausal symptoms in women. Elephant foot yam contains phytoestrogens – isoflavones, coumestans and lignans. A limited research has look at these functional properties of elephant foot yam. It has both nutritional and medicinal value.

Objective: To evaluate and promote the functional properties of Elephant Foot Yam (Amorphophallus paeoniifolious) to ameliorate perimenopausal systems and improve the quality of life in middle age women.

**Methods:** Women (n=100; 50 for experimental group and 50 for control group) between age group of 35-60 years who fulfilled the inclusion and exclusion criteria and given their consent were enrolled. The experimental group (EG) was supplemented with fine powder of yam and control group (CG) was provided nutrition health education for 45 days. Anthropometric measurements and menopause related symptoms score (MRS - using a standard questionnaire from the Professor Heinemann from Center of Epidemiology and Health Studies, Berlin, Germany) of study subjects were obtained before and after supplementation.

**Results:** Mean±SD for weight in EG and CG were 62±7.4 and 63.2±7.4 respectively before intervention; which were reduced significantly to 60.6±6.9 (p<0.001) in EG and non-significantly to 63.1±7.5 (p=0.6) in CG. Similarly for BMI, EG showed a significant

reduction from 25.9±3.4 to 25.3±3.2 (p<0.001) and no
specific change was observed for BMI in control group. Waist cir-
cumference was significantly reduced in EG – from 89.67±4.7 to
88.65±5 (p<0.01).

Mean±SD of MRS in EG was 11.9±2.5, which reduced to 6.3±3 after supplementation (p<0.001). A reduction in mean±SD of MRS was also observed in CG group where 11.2±1.52 was reduced to 9.0±1.7. Mean value showed a marked reduction for all three dimensions in both groups as well.

**Conclusions:** The study supports acceptability and effectiveness of elephant foot yam to improve quality of life in women. Detailed studies on underutilized crops, such as yams, and their health benefits are needed if food security and nutrition security are to be assured.

**Keywords:** Elephant Foot Yam, Menopause Rating Scale (MRS), Phytoestrogen.

144/1928

HYDROLYSED PEA PROTEINS MITIGATE IN VITRO WHEAT STARCH DIGESTIBILITY

(1)Lopez Baron, Nataly; (2)Sagnelli, Domenico; (3)Blenow, Andreas;

(4)Holse, Mette; (5)Saaby, Lasse; (6)Gao, Jun; (7)Vasanthan, Thava.

(1)PhD. Food Chemist. Department of Agricultural, Food and Nutritional Science. University of Alberta. Canada.; (2)PhD. Biotechnologist. Department of Plant and Environmental Sciences. University of Copenhagen. Denmark; (3)PhD. Associate Professor. Department of Plant and Environmental Sciences. University of Copenhagen. Denmark.; (4)PhD. Food Scientist. Department of Food Science. University of Copenhagen. Denmark.; (5)PhD. Pharmacologist. Bioneer. Denmark.; (6)MSc. Senior Lab Tecnologist. Department of Agricultural, Food and Nutritional Science. University of Alberta. Canada; (7)PhD. Professor. Department of Agricultural, Food and Nutritional Science. University of Alberta. Canada.

**Background and objectives:** Isolated pea proteins and their hydrolysates show potential to reduce blood glucose levels. An in vitro amylase digestibility study of cooked wheat starch in the presence of pea protein isolate in their native, heat-denatured and protease-hydrolysed counterparts was conducted in our lab. The results revealed that protein denaturation and protein hydrolysis enhanced starch-protein interaction that may have reduced the rapidly digestible starch content. Nevertheless, prior to a clinical trial, a more realistic in vitro digestibility technique that simulates the human gastrointestinal tract was needed to better understand this effect.

**Objectives:** Determine amylolysis of cooked wheat starch in the presence of native and hydrolysed pea protein by measuring the glucose release, in an in vitro static digestion model.

Determine starch amylolysis of extruded snacks prepared with blends of wheat flour and native or protease hydrolysed pea proteins at 12%, by measuring glucose release in an in vitro gastrointestinal dynamic model.
**Methods:** In vitro static digestion: Cooked wheat starch (control) and blends with native or hydrolysed pea protein, were mixed with simulated salivary fluid at pH6.9 and incubated (37°C-5min-170rpm). Samples were mixed with simulated gastric fluid pH2 for 2h in the presence of pepsin. Then, simulated intestinal fluid pH7 was added in the presence of bile salts and pancreatin. Samples were collected for glucose determination every 20min for 2h.

In vitro dynamic digestion: Extruded samples of wheat flour+native/hydrolysed pea proteins were mixed with human saliva and poured in a fundus resembling the stomach. Acid and pepsin were added gradually. Samples were collected every 7min and mixed in a vessel containing intestinal fluid. After the addition of the last sample from the fundus to the static intestinal digestion, samples were collected every 20min for 2h for glucose release determination.

**Results:** Native and protease-hydrolysed proteins when cooked with starch showed significant reduction in glucose release compared to the control at 40, 60 and 120min.

The addition of hydrolysed-protein to an extruded wheat snack reduced significantly the glucose release at 5, 20, 40 and 90min.

**Conclusions:** The study suggests a possible novel natural protein ingredient to formulate low glycemic food products.

**Keywords:** Pea proteins, Protein-hydrolysis, Starch, In vitro digestibility.

**Conflict of Interest Disclosure:** This study was supported by Natural Sciences and Engineering Research Council of Canada (NSERC), Alberta Innovates BioSolutions and the Colombian Administrative Department of Science, Technology and Innovation (Colciencias).

**Methods:** Amaranth protein was isolated according to the isoelectric precipitation method, obtaining 89% of purity. The assay was performed with twenty-four hamsters divided into three groups with diets distinguished by protein source and fat profile. Experimental group (I) were fed a diet with 23% amaranth protein, high in SAFA and with 0.1% cholesterol. The control groups received a diet with 23% casein and fat profile according to AIN-93 (N), or 23% casein, high in SAFA and with 0.1% cholesterol (H). The total lipid content of livers was measured by gravimetric method, cholesterol concentration was analyzed by HPLC-DAD and fatty acids profile was performed by GC.

**Results:** Group I presented lower total lipid concentration (24%) compared to group H and did not differ from N group. The same occurred to cholesterol concentration in liver (65% lower than H group and similar to N group). Fatty acids profile for I group presented about 53% SAFA, H group presented 52% and N group presented 41%. Monounsaturated fatty acids (MUFA) was 16% for I group, 24% for H group and 9% for N group, furthermore polyunsaturated fatty acids (PUFA) was 32% for I group, 24% for H group and 50% for N group.

**Conclusions:** The use of amaranth protein demonstrated positive changes in liver lipid profile even including high-saturated fat acids in diet. Possibly amaranth peptides are absorbed and achieve liver cells, and/or amaranth protein act in intestine by signaling pathways. Therefore, stimuli on the consumption of amaranth represent an alternative to diversify the diet and promote a reduction of the risk of CVD.

**Keywords:** Amaranth protein, Lipids metabolism, Hypocholesterolemic effect.

144/1943

## AMARANTH PROTEIN ACTS AS A PROTECTIVE FACTOR IN LIVER OF HAMSTERS FED WITH HIGH CONCENTRATIONS OF SATURATED FATTY ACIDS AND CHOLESTEROL

**Background and objectives:** Incidence of cardiovascular disease (CVD) in Western countries is elevated and related to high cholesterol levels, and is evident the role of diet in the control of cholesterol and dyslipidemia. Although studies suggest amaranth protein has a hypocholesterolemic effect, the biological pathways by which the protein acts are still not fully known. Thus the objective of this study was to evaluate the effect of ingestion of amaranth protein over lipid profile in liver of hamsters fed with high concentrations of saturated fatty acids (SAFA) and cholesterol.

144/1949

## EFFECT OF ANTIOXIDANT DIETARY FIBER FROM SPENT COFFEE (COFFEA ARABICA L.) GROUNDS ON GASTROINTESTINAL HEALTH OF ADULTS

**Background and objectives:** Spent coffee grounds (SCG), the residue obtained after brewing, is rich in dietary fiber, compound that imparts several health benefits such as prevention or reduction of bowel disorders, and decrease risk of coronary heart disease and type 2 diabetes. Previously a novelty antioxidant dietary fiber rich extract was obtained from SCG (ADFSCG). Moreover, with higher dietary fiber intake, stool weight tends to be higher and transit time enhances which may contribute to the prevention of large bowel disorders such as constipation, diverticulitis and large bowel cancers. Thus, the objective of this study was to evalu-

<footnote>(¹)Manólio Soares Freitas, Rosana Aparecida; (¹)Caliman Salvador, Bianka; (¹)Pereira da Silva, Cintia; (²)Maria Guizellini, Gloria;</footnote>

<footnote>(³)Rodrigues Sampaio, Geni; (³)Gomes Aréas, José Alfredo.</footnote>

<footnote>(¹)MSc. Nutrition Department. School of Public Health. University of São Paulo – USP. São Paulo. Brazil.; (²)Nutrition student. Nutrition Department. School of Public Health. University of São Paulo – USP. São Paulo. Brazil; (³)PhD. Nutrition Department. School of Public Health. University of São Paulo – USP. São Paulo. Brazil.</footnote>

<footnote>(¹)Gómez Castrejón, Karen Lizzet; (²)Campos Vega, Rocio.</footnote>

<footnote>(¹)Universidad Autónoma de Querétaro, Facultad de Química, Departamento de Investigación y Posgrado de Alimentos (DIPA), Cerro de las Campanas s/n, C.P. 76010, Santiago de Querétaro, Qro. México.; (²)Universidad Autónoma de Querétaro, Facultad de Química, Departamento de Investigación y Posgrado de Alimentos (DIPA), Cerro de las Campanas s/n, C.P. 76010, Santiago de Querétaro, Qro. México.</footnote>
ate the effect of ADFSCG (5g/day) consumption (21 days), as part of a cookie, on gastrointestinal health (43 adults) in a randomized and parallel to a control group (cookie without fiber) trial.

**Methods:** The intestinal habits and stool consistency were determined by Bristol scale, and the severity and frequency of 18 gastrointestinal symptoms were also evaluated (FDA). The enzymatic activity of β-glucosidase, β-glucuronidase, tryptophanase and urease as well as the antioxidant capacity (DPPH) were determined in faeces.

**Results:** None of the cookies significantly modified the anthropometric parameters. After ADFSCG cookies intake, total cholesterol increased significantly (within normal values). This group showed a decrease in the general discomfort, with significantly impact on inflammation. On the other hand, all the 18 gastrointestinal evaluated symptoms were improved after ADFSCG cookies intake, showing a significantly decrease in the frequency of diarrhea; while, a lower retrosternal burning was reported after-control cookie intake. A decrease in β-glucosidase and β-glucuronidase was observed in the ADFSCG group, with no statistical difference in faeces antioxidant capacity (DPPH).

**Conclusions:** The ADFSCG can serve as a source of dietary fiber by improving the bowel's well-being. Additional studies are needed to extend the knowledge of ADFSCG effects on gastrointestinal health.

**Keywords:** Spent coffee grounds, Antioxidant dietary fiber, Gastrointestinal health.

**Further collaborators:**

R. Campos-Vega PhD

144/1958

ENDOSPERM OF TARA SEED (CAESALPINEA SPI-NOSA) AND AMERICAN AGAVE LEAVES POW-DER REGULATE BODY WEIGHT GAIN AND IN-TESTINAL HEALTH IN HOLTZMAN RATS

**Background and objectives:** There is scientific evidence about dietary fiber intake in prevention and treatment of non-transmissible diseases, which effects depend on type and fiber proportion. On this basis, the present study aimed to investigate the effect of different proportions of endosperm of tara seed (ETS), powder of the Agave americana leaves (AAL) and cellulose (pattern) on the physiological properties related to body weight gain and intestinal health.

**Methods:** Holtzman rats were fed with different proportions of ETS:AAL (6:0, 3:3, 0:6, 10:0, 5:5, 0:10 % respectively) and measure of feed intake, gain body weight, fat apparent digestibility, volume and fecal pH.

**Results:** Lower feed intake, decreased body weight gains, fecal mass and pH were showed in ETS (soluble fiber) diets in comparison to control diet. Indeed, higher values in fecal fat content and lower fat apparent digestibility were found to ETS. On the other hand, an increase in fecal volume and decrease in gut transit time were showed in animals feed with AAL (insoluble fiber).

**Conclusions:** These findings demonstrated that ETS reduce body weight gain due to hydration capacity and to the formation of viscous solutions of soluble fiber (75%) in the intestinal lumen that avoid contact between the digestive enzymes and nutrients. AAL powder with higher fiber insoluble content (80%) may play a role in the prevention and treatment of constipation due to its water retention capacity, low viscous solution formation and increase fecal volume.

**Keywords:** Dietary fiber. Soluble fiber. Insoluble fiber. Physiological properties.

**Further collaborators:**

To Fondo Nacional de Desarrollo Científico, Tecnológico y de Innovación Tecnológica con el Convenio de Subvención N° 275-2015-FONDECYT-PERU for the financial support of the study as well as to "Molinos Asociados SAC"-Lima for the donation of ETS.

144/2036

GPETAFLR, A PEPTIDE ISOLATED FROM LU-PINUS ANGUSTIFOLIUS L. PROTEIN HYDRO-LYSATE, EXERTS ANTI-INFLAMMATORY EF-FECTS IN HUMAN PRIMARY MONOCYTES

**Background and objectives:** Recent studies have shown that lupine protein hydrolysates (LPH) could be an important source of bioactive peptides, especially showing anti-inflammatory activity. The present study aimed to test whether peptide GPETAFLR released from the enzymatic hydrolysis of lupine protein may modulate the inflammatory response in human primary monocytes.

**Methods:** Freshly human monocytes were used to analyze the effects of GPETAFLR, after 24 hours of treatment at 50-100 µg/ mL, on LPS-induced inflammatory response using FACS analysis, RT-qPCR, and ELISA procedures.

**Results:** We report that GPETAFLR from LPH possesses anti-inflammatory properties in human monocytes. GPETAFLR skewed the monocyte plasticity towards the anti-inflammatory non-classical CD14+CD16++ monocytes and reduced the inflammatory competence of LPS-treated human monocytes diminishing IL-1β, IL-6, and TNF-α production and gene expression. **Results:** **Conclusions:** Taken together, our results show that

<footnote>(1) Vilcanqui Pérez Fulgencio; (2) Villanueva Espinoza, María Elena;</footnote>

<footnote>(3) Vilchez Perales, Carlos.</footnote>

<footnote>(1) MSc. Departamento de Nutrición. Universidad Nacional Agraria la Molina. Perú; (2) Dra. Departamento de Nutrición. Universidad Nacional Agraria la Molina. Perú; (3) Dr. Departamento de Nutrición. Universidad Nacional Agraria la Molina. Perú.</footnote>

<footnote>(1) Millán-Linares, Mª Carmen; (1) Montserrat de la Paz, Sergio; (1) Yust Escobar, Mª del Mar; (1) Pedroche Jiménez, Justo Javier; (2) Millán Rodríguez, Francisco.</footnote>

<footnote>(1) Research, Health and Nutrition Department, Instituto de la Grasa (CSIC), Sevilla, España; (2) Research Professor, Health and Nutrition Department, Instituto de la Grasa (CSIC), Sevilla, España.</footnote>
GPETAFLR has the potential to impair the activation of human primary monocytes and the derived inflammatory conditions, and suggest a new role for Lupinus angustifolius L. protein hydrolysate in the regulation of the pathogenesis of health disorders in which monocytes play a key role, including atherosclerosis and several chronic diseases.

**Keywords:** Lupine, Bioactive peptide, Protein hydrolysates, Monocyte, Inflammation.

**Further collaborators:**

Laboratory of Cellular and Molecular Nutrition, Instituto de la Grasa (CSIC)

Research Professor Fco. J. García Muriana

144/2070

THE ANTIOXIDANT DIETARY FIBER FROM SPENT COFFEE (COFFEA ARABICA L.) GROUNDS IMPROVE CIRCADIAN LOCOMOTOR ACTIVITY AND ANTHROPOMETRIC PARAMETERS IN ADULTS WITH NORMAL WEIGHT AND OVERWEIGHT

(1)Campos Vega, Rocio; (2)Oseguera Castro, Karla Yadhira; (3)García Obregón, Olga Patricia; (4)Loarca Piña, Guadalupe; (4)Mendoza Díaz, Sandra Olimpia.

(1)PhD. Programa en Alimentos del Centro de la República (PROPAC). Facultad de Química. Universidad Autónoma de Querétaro. México.; (2)BsC. Programa en Alimentos del Centro de la República (PROPAC). Facultad de Química. Universidad Autónoma de Querétaro. México.; (3)PhD. Programa de Nutrición. Facultad de Ciencias Naturales. Universidad Autónoma de Querétaro. México.; (4)PhD. Programa en Alimentos del Centro de la República (PROPAC). Facultad de Química. Universidad Autónoma de Querétaro. México.

**Background and objectives:** All organisms present processes in their physiological functions that occur in a periodic and predictable way called "biological rhythms", including circadian rhythms (CR). Disruption of these rhythms (chronodisruption) is associated with an increased risk to develop non-communicable diseases such cancer, overweight and obesity, specially in the evening chronotype, for which reason the establishment of appropriate circadian rhythms is a priority. Recently, foods have been proposed as a powerful modular of the CR, however there is no evidence of the effect of fiber consumption on chronodisruption parameters like locomotor activity. The aim of this study was to evaluate the effects of the consumption (21 days) of antioxidant dietary fiber (ADFSCG, 5g/day), extracted from spent coffee grounds (SCG), as part of a cookie (C-ADFSCG), on the circadian locomotor activity, plasma antioxidant capacity as well as biochemical parameters of adults with normal weight and overweight.

**Methods:** The study was randomized and parallel to a control group (cookie without fiber).

**Results:** Caloric intake decreased in subjects who consumed the C-ADFSCG, reflected on the decrease of waist circumference

and body fat percentage, in relation to baseline values and to the control group (p <0.05). In addition, plasma antioxidant capacity increased significantly (p <0.05) after C-ADFSCG consumption. Normal weight subjects (36%, from both treatments) increased their sleep quality and physical activity. At the beginning of the study, morningness chronotypes in subjects with normal weight, and eveningness chronotype in subjects with overweight, were identified. Interestingly, a significant increase in morningness chronotypes was observed in the 62.5% of overweight subjects and 45.4% on the normal weight subjects (63% of this group population) after C-ADFSCF consumption.

**Conclusions:** This study informs, for the first time, the potential of antioxidant dietary fiber, particularly ADFSCG, to modulate circadian locomotor activity, plasma antioxidant capacity and body fat.

**Keywords:** Spent coffee grounds, Antioxidant dietary fiber, Circadian rhythms, Plasma antioxidant capacity.

**Further collaborators:**

PhD. Rocio Campos Vega. Programa en Alimentos del Centro de la República (PROPAC). Facultad de Química. Universidad Autónoma de Querétaro. México.

144/2093

NUTRITIONAL, BIOACTIVE, AND BOTANICAL CHARACTERIZATION OF BEE POLLEN HARVESTED IN THE COLORADO RIVER VALLEY, ARGENTINA

(1)García Paoloni, María Soledad; (2)Vallese, Federico Danilo;

(3)Springer, Valeria; (4)Andrada, Ana Cristina; (3)Pistonesi, Marcelo Fabián.

(1)Biochemistry. Laboratorio de Apicultura. INTA Hilario Ascasubi. Argentina; (2)BSc. INQUISUR. Departamento de Química. Universidad Nacional del Sur (UNS)-CONICET. Argentina; (3)PhD. INQUISUR. Departamento de Química. Universidad Nacional del Sur (UNS)-CONICET. Argentina; (4)PhD. Dpto. Agronomía. Universidad Nacional del Sur (UNS). Argentina.

**Background and objectives:** Honey bees (Apis mellifera L.) collects pollen from flowers as pollen loads to get proteins for sustaining the colony. Due to its protein content (macronutrient) bee pollen is consumed as a nutritional supplement. Additionally the presence of bioactive compounds like polyphenols (micronutrients) provides biological function reason why this bee product is considered a natural functional food. Bee pollen composition depends strongly on the botanical and geographical origin (even though post-harvested processing). The aim of this study is to characterize bee pollen collected in the Buenos Aires Valley of Colorado River (BAVCR) zone.

**Methods:** During the beekeeping season 2013-2014, twelve samples were harvested to quantify total proteins and total polyphenols and to evaluate the botanical origin. Total protein content was estimated by Kjeldahl method with a conversion factor of 6.25 as the Argentine Food Code indicates (Chapter X, art.
785) and the mean value obtained was 22% ± 2%. Total polyphenols was determinate by the reference method of Folin-Ciocalteu and the average content was 13.1 ± 3.3 mg gallic acid/g pollen.

**Results:** The values obtained are comparable to those reported in the literature. The botanical origin was established by acetolysis method (Erdtman-1960) and subsequently the pollen grains were identified and counted using light microscope. Main represented taxa in the samples were Brassicaceae, Lotus and Eucalyptus.

**Conclusions:** Both protein and polyphenols obtained values allow us to conclude that bee pollen collected in the BAVCR can be considered an excellent dietary supplement as well as a natural functional food. The vegetation availability of mentioned botanical taxa will enable beekeepers of this geographical area to obtain very good quality bee pollen.

**Keywords:** Functional food, Dietary supplement, Total protein, Polyphenols, Botanical origin.

Count agar + nistatina while the count of mould/yeast was done in Sabouraud dextrose agar at 26°C.

**Results:** Fifty percent of BP loads from both controls showed fungal growth after 48 hours of incubation while treatments with PEE developed growth after 168 hours (seven days). The same results were obtained when antibacterial activity was measured on BP loads from PEE treatments. The total of the BP loads from controls were contaminated by bacteria and fungus after 72 hs. In contrast, PEE5 and PEE7.5 never reached this percentage during the whole trial.

**Conclusions:** This study allowed us to conclude that PEE present antimicrobial activity against bacteria and mould/yeast from fresh BP loads. Thus, PEE becomes a new alternative that offers a preservation method for BP intended for human consumption. Further investigations may reveal extra nutritional power conferred by PEE.

**Keywords:** Functional food, Nutritional supplement, Microbiological food preservation.

144/2109

## PROPOLIS ETHANOLIC EXTRACT AS NATURAL BEE POLLEN LOAD PRESERVATIVE

(1)García Paoloni, María Soledad; (2)Krepper, Gabriela; (3)Cibanal, Irene; (4)Fernandez, Leticia; (5)Centurión, María Eugenia; (5)Di Nezio, María Susana; (5)Pistonesi, Marcelo Fabián.

(1)Biochemistry. Laboratorio de Apicultura. INTA Hilario Ascasubí. Argentina; (2)Biochemistry. INQUISUR Departamento de Química. Universidad Nacional del Sur (UNS)-CONICET. Argentina; (3)Agricultural engineer. Laboratorio de Estudios Apícolas (LabEA). Universidad Nacional del Sur (UNS)-CIC. Argentina;

(4)PhD. Laboratorio de Estudios Apícolas (LabEA). Universidad Nacional del Sur (UNS)-CONICET. Argentina; (5)PhD. INQUISUR Departamento de Química. Universidad Nacional del Sur (UNS)-CONICET. Argentina.

**Background and objectives:** Bee-products are recognized worldwide as natural functional foods mainly owing to their polyphenols content. Additionally there are many scientific reports about the use of propolis ethanolic extracts (PEE) for food conservation. In Argentina bee pollen (BP) is used as a dietary supplement and quality parameters are regulated in the Argentine Alimentary Code (Chapter X, art. 785). The drying process prior to commercialization is critical to prevent microbiological development and therefore increase larger shelf life. However, the temperature of this step must be controlled to avoid the loss of the phenolic components. The aim of this work was to determine the antimicrobial activity of PEE application over BP loads.

**Methods:** The antimicrobial effect against bacteria and yeast/ mould was tested separately using the percentage of contaminated pollen loads method. For this in vitro trial, 20 Petri plates (20 BP loads per plate) were divided in four treatments with five repetitions: one control with BP loads (T1); another control with BP loads soaked with 7.50 µL of ethanol (T2), and BP loads soaked with 5.00 µL (PEE5) and 7.50 µL (PEE7.5) of PEE respectively. The number of pollen loads with bacteria was studied at 35°C in Plate

144/2132

## POLYPHENOLS CONSUMPTION AND EXPRESSION OF GENES RELATED TO ENDOPLASMIC RETICULUM STRESS IN VASCULAR ENDOTHELIUM

(1)Meza Miranda, Eliana; (2)Rangel Zúniga, Oriol; (3)Haro, Carmen;

(3)García Carpintero, Sonia; (4)García Ríos, Antonio; (4)López Miranda, José; (4)Pérez Jiménez, Francisco; (5)Camargo, Antonio.

(1)MSc. PhD. Nutricionist. Centro Multidisciplinario de Investigaciones Tecnológicas. Universidad Nacional de Asunción. Paraguay; (2)PhD. Microbiologist. Lipids and Atherosclerosis UNIT, IMIBIC, CIBEROBN; (3)PhD. Biologist. Lipids and Atherosclerosis UNIT, IMIBIC, CIBEROBN; (4)PhD. MD. Lipids and Atherosclerosis UNIT, IMIBIC, CIBEROBN; (5)PhD. Biochemist. Lipids and Atherosclerosis UNIT, IMIBIC, CIBEROBN.

**Background and objectives:** There is a relationship between Metabolic Syndrome (MetS), and expression of genes related to endoplasmic reticulum stress. This condition alter the homeostasis of the endoplasmic reticulum in a chronic manner, which affects normal cellular response. OBJECTIVE: to evaluate the expression of the XBP1, CALR, CANX, HSPA5 and GSTP1 genes after the consumption of a breakfast with virgin olive oil (VOO) rich in phenolic compounds (FC) in HUVEC cells as an in vitro model of vascular endothelium.

**Methods:** Two breakfasts were administered with VOO with high (398 ppm) and low (70 ppm) FC content to 20 patients with MetS in a double-blind, randomized and crossover design. The HUVEC were treated with serum obtained at 0,2 and 4 hours after the consumption of the breakfasts. Gene expression was performed with OpenArray (Applied Biosystem).

**Results:** In comparison to serum obtained in the fasting state, treatment of HUVEC with serum obtained after consumption of VOO low in FC, decreased expression of XBP1, HSPA5 and CANX genes (p= 0.03, p=0.02 and p=0.01 respectively) at 2 hours
post-breakfast intake and increased expression of CALR gene (p = 0.03) at 4 hours post-breakfast intake. The treatment of HUVEC with serum obtained after consumption of the VOO-prepared breakfast with high FC content, on the other hand, decreased the expression of XBP1, CALR and GSTP1 genes (p=0.02, p=0.01, and p=0.05 respectively) at 4 hours post-breakfast intake. The comparison between treatments showed a significantly lower expression of XBP1, CALR and GSTP1 genes (p <0.001, p<0.001 and p=0.03 respectively) after the intake of the breakfast prepared with VOO with high content of FC at 4 hours in the postprandial state.

**Conclusions:** VOO rich in FC intake, regulates the expression of genes related to endoplasmic reticulum stress in patients with MetS in the postprandial state, which contributes to a less activation of cellular stress signaling.

**Keywords:** Phenolic compounds, Metabolic syndrome, Vascular endothelium, Endoplasmic reticulum stress.

144/2135

APPLICATION OF PEAR ORANGE CITRUS FOOD FIBER (CITRUS SINENSIS OSBECK) IN THE PRODUCTION OF PRODUCTS FUNCTIONAL FOODS

**Background and objectives:** Consumers often despise parts of fibers of the vegetables. The use of by-products of fruits adds value to new food products. OBJECTIVE: Develop cakes with flour of flavedo, albedo and pear orange pulp (FAP) and characterize them sensorially.

**Methods:** At the Laboratory of Food of Plant Origin of the Federal University of Rio de Janeiro the pear orange was submitted to the hygiene and extraction of the juice. The residue was dehydrated and crushed to obtain the citric flour. Four cakes were prepared, 3 of which were added with 1.5; 3.0 and 5% of citrus flour, respectively, and 1 cake without citrus flour. Common ingredients of cakes: vegetable oil, demerara sugar, egg, baking powder, wheat flour and orange juice. The cakes were submitted to an Acceptance Test by Order of Preference in a scale of preference, preferably in level 4, and the least in level 1. The statistical test was based on Friedman's Order Sum Test at 1% probability. The research was submitted to the Research Ethics Committee and its approval protocol is 954,908 with application to the Free and Informed Consent Form.

**Results:** 49 volunteers between 18 and 53 years old accomplish the Preference Ordination Test. There was no significant difference between the cakes of 0; 1.5; 3.0 and 5% of citrus flour at the 1% probability level, demonstrating that the tasters did not measure

the distinction between these samples. Among the 0% and 5% samples, there was a statistical difference at the 1% probability level. The cake with 5% of citrus flour was the less preferred, which can be explained by the higher percentage of citric fiber, which may interfere with the taste and texture of food products.

**Conclusions:** Citrus flour in cakes seems to be good for the use of pear orange by-products due to their good acceptance, who did not present significant differences in the sensory acceptance test and can be consumed by the general population.

**Keywords:** Dietary fiber, Citric fiber, Orange-pear, Soluble fiber, By-products.

**Conflict of Interest Disclosure:** The authors below declare that they participated in the conception, analysis of results and contributed effectively to the accomplishment of the Scientific Summary: " Application of pear orange citrus food fiber (Citris sinensis Osbeck) in the production of products functional foods". They make public the responsibility for its content, that no links or financing agreements between authors and companies that may have interest in the publication of this Scientific Summary have been omitted. They affirm that they have no conflict of interest with the topic addressed in the article, nor with the products cited. We declare that the above Scientific Summary is original and that the work, in part or in full, or any other work with substantially similar content, has not been submitted to and will not be submitted to the other Scientific Congress as long as its publication is being considered. List of authors: Simone Silveira van Boekel Alexandre Marques van Boekel (CPF 69984930700). Lais Buriti de Barros (CPF 00489553702).Déborah Carolina Martins de Jesus (CPF 14685199723). Priscila Vieira Pontes (CPF 08307924731). Larissa Ribeiro Barreta Matheus (CPF 13532972760). Nicolly Rindolphy Basilio Rodrigues (CPF 135019958716).

144/2146

ANTIOXIDANT ACTIVITY AND SENSORY ANALYSIS OF FROZEN KALE PULP (BRASSICA OLERACEA L. VAR. ACEPHALA)

**Background and objectives:** Kale (Brassica oleracea L. var. Acephala) is a vegetable whose consumption has increased due to researchs on its nutraceutical properties. This vegetable is rich on natural antioxidant compounds and have high levels of carotenoids, ascorbic acid, flavonoids, tocopherols, phenolic compounds

<footnote>(1)Van Boekel, Simone; (2)Rodrigues, Nicolly; (3)Jesus, Déborah;</footnote>

<footnote>(3)Matheus, Larissa; (4)Buriti, Lais; (4)Pontes, Priscila.</footnote>

<footnote>(1)PhD. Nutritionist. Teacher. Course of Nutrition. Federal University of Rio de Janeiro.Campus Macaé. Rio de Janeiro.;</footnote>

<footnote>(2)Graduate Course Nutrition. Federal University of Rio de Janeiro. Campus Macaé. Brazil.; (3)Nutritionist. Rio de Janeiro. Brazil.;</footnote>

<footnote>(4)Teacher of Nucleous of Food Technology. Federal University of Rio de Janeiro.Campus Macaé. Brazil.</footnote>

<footnote>(1)Mastella, Paula; (2)Fabregat, Mariane; (2)GEBARA, TELMA; (3)Souza, Wesley; (4)Iwamura, Louise; (5)Tarzia, Andréa.</footnote>

<footnote>(1)PhD. Department of Biological and Health Sciences. Tuiuti University of Parana. Brazil.; (2)BSc. Nutritionist. Department of Biological and Health Sciences. Tuiuti University of Parana. Brazil.; (3)BSc. Department of Biological and Health Sciences. Tuiuti University of Parana. Brazil.; (4)MSc. Nutritionist. Department of Biological and Health Sciences. Tuiuti University of Parana. Brazil.; (5)PhD. Nutritionist. Department of Biological and Health Sciences. Tuiuti University of Parana. Brazil. andreatarzia@hotmail.com.</footnote>
and glycosinolates, substances that act as antioxidants and that protect the human body against reactive oxygen species. Most perishable foods can be preserved by refrigeration and freezing for a limited time, with the benefits of retarding microbial activities and retaining their nutrients. The objective of this research was to elaborate kale pulp and evaluate its antioxidant activity and sensory acceptability after different periods of freezing.

**Methods:** Samples (6 kg of organic kale leaves) were cleaned, sanitized and processed in order to obtain as final product the kale pulp. Samples were frozen (-18°C) and analyses were performed after 5, 30, 60 and 75 days of storage under freezing. An affective (hedonic) sensory test was performed on each test day by untrained assessors. For the affective test the taste was investigated on a 5-point hedonic scale. Antioxidant capacity of the products were evaluated according to the method of Prieto et al. (1999). Antioxidant capacity of the pulps were expressed as relative antioxidant activity (RAA%) to ascorbic acid. Results were analyzed using analysis of variance (ANOVA), and mean values were compared using Tukey's test. Values of p ≤0.05 were considered statistically significant. The data were analyzed using Statistica 8.0 (Statsoft Inc., Tulsa, OK, USA).

**Results:** Results indicated that freezing the kale pulp up to 30 days did not affect the product acceptability by assessors regarding the parameter analyzed (taste). No difference (p<0.05) was found on RAA% during the tested period, and values ranged from 9.37% (Day 5) to 8.87% (Day 75), showing a good retention of the original antioxidant activity at the end of storage time.

**Conclusions:** Freezing of kale pulp can be seen as a good alternative for preserving the vegetable quality for a period of 30 to 60 days, without significant losses in the antioxidant activity and sensory profile.

**Keywords:** Brassica oleracea L. var. acephala, Kale pulp, Hedonic scale, Antioxidant activity.

**Conflict of Interest Disclosure:** Brassica oleracea L. var. acephala, kale pulp, hedonic scale, antioxidant activity.

researchers for its therapeutic and nutritional properties. Prepared as fruit pulp juice has great demand in alternative medicine for the treatment of diseases and there are many works that prove its pharmacological properties. However, there are few scientific studies on the seeds of the fruits (Barros, 2009). The most common bioactive compounds determined in all parts of plants are the phenolic substances, which are formed in the secondary metabolism of plants. These compounds can be found in free form or linked to sugars and proteins, having several functions (Liu, 2007). The objective of this work was to analyze the content of bioactive compounds, vitamin C, titratable acidity and antioxidant activity of noni seeds.

**Methods:** The noni seeds were separated from the fruit pulp and dried in an oven at 60 °C for 12 hours. Vitamin C analysis by titration with 2,6-dichlorophenolindofenol sodium solution, determination of phenolic compounds by the Follin-Ciocauteau method, carotenoids, total flavonoids and antioxidant activities by free radical capture DPPH (2,2-Diphenyl-1 -pyryl-hydrazyl).

**Results:** The pH value of the dried seed was 4.97. The total acidity resulted in 0.006 g / 100g. The vitamin C content was 303.03 mg / 100g. The content of phenolic compounds obtained was 297.00 mg equivalent of gallic acid / 100 g. The flavonoid content was 25.04 mg / 100 g and carotenoids 15.16 mg / 100 g. The antioxidant activity resulted in 90.96%. With these results we can highlight the high content of vitamin C and great potential of antioxidant activity. According to Costa et al. (2013) noni is a representative fruit with significant amounts of vitamin C.

**Conclusions:** The noni seed presented low content of organic acids, content of phenolic compounds such as flavonoids and carotenoids, high content of vitamin C and with great potential of antioxidant action.

**Keywords:** Morinda citrifolia L., Seeds, Bioactive compounds, Antioxidant.

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ANALYSIS OF BIOACTIVE COMPOUNDS OF NONI SEEDS (MORINDA CITRIFOLIA L.)

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

(1)Dra. Agronomy. Food Technology. Federal Institute of Mato Grosso do Sul. Coim. Brazil.; (2)Graduate student of Food Technology. Federal Institute of Mato Grosso do Sul. Coxim, Brazil.; (3)Graduate Student in Chemistry. Federal Institute of Mato Grosso do Sul. Coxim. Brazil.; (4)Graduate Student in Food Technology. Federal Institute of Mato Grosso do Sul. Coxim. Brazil.; (5)Dr. Vegetable Production. Federal Institute of Mato Grosso do Sul. Coxim. Brazil.

**Background and objectives:** The noni (Morinda citrifolia L.) is a plant that has a fruit that has aroused the interest of the

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EFFECT OF LACTOBACILLUS FERMENTUM CRL1446 ON IMMUNE-METABOLIC PARAMETERS AND INTESTINAL MICROBIOTA COMPOSITION IN OBESITY IN HIGH-FAT DIET FED MICE

(1)Gauffin-Cano, María Paola; (2)Fabersani, Emanuel; (3)Marquez, Antonela; (4)Ross, Romina; (2)Abeijón-Mukdsi, Claudia; (1)Medina, Roxana.

(1)PhD. Biochemist. Centro de Referencia para Lactobacilos (CERELA). CONICET. Argentina; (2)PhD. Biotechnologist. Centro de Referencia para Lactobacilos (CERELA). CONICET. Argentina; (3)BSc. Biotechnologist. Centro de Referencia para Lactobacilos (CERELA). CONICET. Argentina; (4)PhD. Pharmacist. Centro de Referencia para Lactobacilos (CERELA). CONICET. Argentina.

**Background and objectives:** Dietary strategies, including the use of probiotics as preventive agents that handle the intestinal microbiota and/or modulate the function of adipose tissue, would be suitable tools for the prevention or treatment of different condi-
tions associated with obesity. In this context, our aim was to evalu-
ate the effect of Lactobacillus fermentum CRL1446 (CRL1446) on
metabolic and immunological parameters and on the composition
of intestinal microbiota in a model of obesity induced by a high-fat
diet (HFD).

**Methods:** Balb/c mice were separated into the following groups: 1) SD: Standard diet, 2) HFD: High-fat diet, 3) HF-D+CRL1446: mice receiving a HFD and a daily dose of 1 x 108 CFU of L. fermentum CRL1446 (CRL1446) in the drinking water. The dietary intervention was maintained for 6 weeks. Weight and food intake were monitored daily. It were determined at 20 and 45 days of experimental period: fat tissue weight (histologic studies), metabolic parameters (triglyceride (TG), total cholesterol (TC), glucose (Glu) and leptin (Lep)), seric immunologic parameters (TNF-alpha, IL-6, IL10 and MCP-1), macrophage and adipocyte functionality (ex vivo studies) and the gut microbiota composition (evaluated by high-throughput sequencing of 16S rRNA gene amplicons).

**Results:** CRL1446 strain induces a decrease in body and adipose tissue weights and the size of adipocytes. A decrease in plasmatic glucose, cholesterol, triglycerides, leptin, TNF-alpha and IL-6 levels and an increase of IL-10 were observed. Ex vivo studies showed a minor secretion of leptin by adipocytes. In macrophages, CRL1446 induced lower levels of MCP-1, TNF-alpha, IL-6 and higher levels of IL-10. Regarding intestinal microbiota, an increase in Lactobacillus and a decrease in Firmicutes/Bacteroidetes (F/B) index were observed.

**Conclusions:** The supplementation with L. fermentum CRL1446 produce changes in intestinal microbiota structure and metabolic and immunologic parameters. CRL1446 could potentially be a probiotic strain useful to correct the immuno-metabolic alterations associated with obesity through its adipomodulatory capacity.

**Keywords:** Obesity, Lactobacillus fermentum, Probiotics, Intestinal microbiota.

of diabetes. Inhibitors of the carbohydrate-hydrolysing enzymes
such as acarbose decrease post-prandial hyperglycaemia. Howev-
er, this drug has certain adverse effects such as diarrhoea and nau-
sea. Therefore, natural inhibitors from foods could be a good strat-
egy to control glycaemic. The Brazilian Savannah has an extensive
biodiversity, but it is underexplored. The jatobá-do- cerrado (Hy-
menaea stigonocarpa Mart.) is a native legume of Brazilian Savan-
nah with great potential for exploration because it's an excellent
source of dietary fiber and polyphenols. The objective of this study
was to evaluate the potential inhibitory activity of jatobá extracts
after digestion in vitro on α-amylase and α-glucosidase enzymes.

**Methods:** The phenolic extract was obtained by sequential ex-
traction with 80% acetone and 60% ethanol. The digested extract
was done using enzyme solutions (α-amilase, pepsin and pancre-
atin) at physiological pH. The total phenolic content (TPC) was
evaluated by Folin reagent. Different concentrations of extracts
were tested.

**Results:** The extract of jatobá after digestion in vitro presents 10 mg GAE.g-1 of sample. Our study have showed that the phenolic extracts greatly inhibited the α-amylase in a positive dose-dependence way (76 and 91% for 9 and 15 mg. mL-1, respectively). On the other hand, the extract showed a high α-glucosidase inhibitory activity (77 and 53%) but this activity is not positively related to concentration of extract (3 and 6 mg. mL-1, respectively).

**Conclusions:** The results of this present study indicate that polyphenols from jatobá are effective α- amylase and α- gluco- sidase inhibitors, which may modulate the postprandial glucose levels. Thus, the jatobá can be helpful in the management of type 2 diabetes.

**Keywords:** Jatobá-do-cerrado, Type 2 diabetes, α-amylase, α-glucosidase.

144/2162

INHIBITION OF ACTIVITY OF CARBOHY-
DRATE-HYDROLYSING ENZYMES BY POLYPHE-
NOLS OF JATOBÁ-DO-CERRADO (HYMENAEA
STIGONOCARPA MART.)

(1)Pereira da Silva, Cintia; (2)Aparecida Manólio Soares Freitas, Rosana; (3)Rodrigues Sampaio, Geni; (4)Alfredo Gomes Arêas, José.

(1)MSc. Nutritionist. Nutrition Department. School of Public Health. University of Sao Paulo. São Paulo. Brazil; (2)MSc. Chemistry. Nutrition Department. School of Public Health. University of Sao Paulo. São Paulo.Brazil; (3)PhD. Biologist. Nutrition Department. School of Public Health. University of Sao Paulo. São Paulo. Brazil; (4)PhD. Professor. Nutrition Department. School of Public Health. University of Sao Paulo. São Paulo. Brazil.

**Background and objectives:** Nutritional management of blood glucose levels is a strategy for the prevention and control

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EVALUATION OF THE ABILITY OF FERMENTED
CACTUS PEAR JUICE TO IMPROVE OXIDATIVE
STRESS TOLERANCE IN YEAST

(1)Torres, Sebastian; (2)Veron, Hernan; (3)Isla, Maria Ines; (4)Gauffin-Cano, María Paola; (5)Dantur, Karina; (6)Fernández-Espinar, María Teresa; (7)Gil-Ponce, Jose Vicente.

(1)PhD. Biochemist. Instituto de Bioprospección y Fisiología Vegetal (INBIOFIV). CONICET. Argentina; (2)BSc. Biotechnologist. Instituto de Bioprospección y Fisiología Vegetal. CONICET. Argentina.; (3)PhD. Biotechnologist. Instituto de Bioprospección y Fisiología Vegetal. CONICET. Argentina.; (4)PhD. Biochemist. Centro de Referencia para Lactobacilos (CERELA). CONICET. Argentina; (5)PhD. Biochemist. Instituto de Tecnología Agroindustrial del NOA. CONICET. Argentina.; (6)PhD. Biologist. Instituto de Agroquímica y Tecnología de Alimentos. CSIC. España.; (7)PhD. Instituto de Agroquímica y Tecnología de Alimentos. CSIC. España.

**Background and objectives:** Cactus pears from Opuntia ficus-indica are strongly recommended in the human diet since
their nutritional and health-promoting features. Scientific evidence has demonstrated the benefits from cactus pear ingestion with special consideration about potentially active antioxidant phytochemicals. However, long-term storage represents the main drawback to the commercialization of this fruit. Since in many places cactus pears are consumed as juice, different procedures were developed to prevent its spoilage. Lactic acid fermentation of cactus pear juice (CPJ) using autochthonous bacteria constitutes an important biotechnology for the exploitation of this fruit. The aim of this study was to evaluate the effect of fermented CPJ (F-CPJ) on oxidative stress tolerance of Saccharomyces cerevisiae.

**Methods:** Lactobacillus plantarum S-811, an autochthonous lactic acid bacteria strain isolated from O. ficus-indica fruits from Northwest Argentina was used to fermented the juice. To evaluate the antioxidant response induced by CPJ and F-CPJ, cultures of S. cerevisiae were incubated with the juices (amounts equivalent to 50, 250 and 500 mg/L of phenolics) during 18 h at 28 °C. Cells were then exposed to sub-lethal oxidative stress, using 0.5 or 4 mM H2O2. After stress treatments, cells were cultured in YPD medium and incubated for 18 h at 28 °C. To evaluate the oxidant effect in each culture, growth-ratio curves (GRC) and effect curves (EC) were constructed as follows: RC = quotient between the growth curve (GC) of the culture exposed to oxidant and the GC of the non-exposed culture; EC = GRC for the culture pre-incubated with the juice was divided by the GRC for the culture pre-incubated without juice.

**Results:** Against a final concentration of 0.5 mM H2O2 there was not observed a significant protective effect by any of the evaluated juices. However, against 4 mM H2O2, CPJ and F-CPJ showed a significant protective effect, that was much higher with the F-CPJ (up to 15 times protection) than CPJ (up to 4 times protection).

**Conclusions:** These results suggest that F-CPJ with L. plantarum S-811 could be a suitable choice for the development of a CPJ with improved functional properties.

**Keywords:** Opuntia ficus-indica, Lactobacillus plantarum, Cactus pear juice.

with the highest concentrations of fiber are flavedo, albedo and pulp most despised. Avoiding waste is sustainability and it is necessary to encourage the full use of fruits, which benefits human beings and the environment. OBJECTIVE: The objective of the present work will be to elaborate cereal bars with flavedo flour, albedo and pulp of orange-pear (FAP) and to evaluate their acceptance through affective sensorial methods.

**Methods:** In the Laboratory of Food of UFRJ, the oranges were sanitized, and then the seeds and the juice were despised. The FAP residue was dehydrated and crushed to obtain the citrus flour. After, were preparation 2 cereal bars of which were added with 3.0 and 5% of citrus flour, respectively. The sensorial analysis of the elaborated bars, was through Acceptance and Preference Tests of 50 non-trained adults participated, after consent in the research that was approved by the Research Ethics Committee.

**Results:** For cereal bars at 3.0 and 5.0% with FAP flour, the note indicated by the non-trained adults in the appearance attributes (25%, respectively); Aroma (45 and 40%, respectively); Texture (20%, respectively); Sweet taste (30%, respectively) and orange flavour (30 and 35%, respectively) was 8.0 indicating a similarity of acceptance between the different attributes. It was stated that up from 30% of non-trained adults buy cereal bars. The t-test with p-value 5% indicated for all attributes, averages very close and p value above 0.05, demonstrating that there was no significant difference between acceptance of samples for all attributes.

**Conclusions:** The sensorial tests showed good acceptance and purchase intention for the two bars with 3 and 5% of FAP flour of orange-pear. In this way, cereal bars may be nutritionally suitable food product options, since they are made with the orange parts that concentrate the dietary fibers.

**Keywords:** Food fiber, Citric fiber, Orange residue, Flavedo, Albedo.

**Conflict of Interest Disclosure:** The authors below declare that they participated in the conception, analysis of results and contributed effectively to the accomplishment of the Scientific Summary: "Application of pear orange citrus food fiber (Citrus sinensis Osbeck) in the production of products functional foods". They make public the responsability for its content, that no links or financing agreements between authors and companies that may have interest in the publication of this Scientific Summary have been omitted. They affirm that they have no conflict of interest with the topic addressed in the article, nor with the products cited. We declare that the above Scientific Summary is original and that the work, in part or in full, or any other work with substantially similar content, has not been submitted to and will not be submitted to the other Scientific Congress as long as its publication is being considered.

List of authors: Simone Silveira van Boekel Alexandre Marques van Boekel (CPF 69984930700). Lais Buriti de Barros (CPF 00489553702). Priscila Vieira Pontes (CPF 08307924731). Camila Mussi Ferreira (CPF 14963874733). Lorrene Rodrigues Pimentel (CPF 16028105759). Juliana de Souza Dofini (CPF 12361030764). Stefani Ribeiro Chaves (15637510709).

<footnote>(1)Van Boekel, Simone; (2)Pimentel, Lorrene; (2)Dofini, Juliana;</footnote>

<footnote>(2)Ferreira, Camila; (2)Chaves, Stefani; (3)Buriti, Lais; (3)Pontes, Priscila.</footnote>

<footnote>(1)PhD. Nutritionist. Teacher. Course of Nutrition. Federal University of Rio de Janeiro.Campus Macaé. Rio de Janeiro.;</footnote>

<footnote>(2)Graduate Course Nutrition. Federal University of Rio de Janeiro. Campus Macaé. Brazil.; (3)Teacher of Nucleous of Food Technology. Federal University of Rio de Janeiro.Campus Macaé. Brazil.</footnote>
(1)Albarracín, Sonia Luz; (2)Velasco, José María; (3)Osorio, Coralia;

(4)Sutachan, Jhon-Jairo.

(1)PhD. Chemistry. Department of Nutrition and Biochemistry. Faculty of Sciences. Pontificia Universidad Javeriana. Bogotá. Colombia; (2)BSc. Nutritionist. Department of Nutrition and Biochemistry. Faculty of Sciences. Pontificia Universidad Javeriana. Bogotá. Colombia; (3)PhD. Chemistry. Department of Chemistry. Faculty of Sciences. Universidad Nacional de Colombia. Bogotá. Colombia; (4)PhD. Physiology and Neuroscience. Department of Nutrition and Biochemistry. Faculty of Sciences. Pontificia Universidad Javeriana. Bogotá. Colombia.

**Background and objectives:** Neurodegenerative diseases such Alzheimer, Parkinson and Huntington are characterized by protein aggregates, mitochondrial impairment, and oxidative stress that can lead to neuronal and astrocytic cell death. Epidemiological studies suggest that consumption of fruits rich in anthocyanins may reduce the risk or delay the development of some neurodegenerative diseases. Bactris guineensis (Corozo) and Pourouma cecropiifolia (Uva caimarona) are endemic plants from the Caribbean and Amazon regions whose fruits are consumed in juices and jellies. Previous studies have reported that these fruits are rich in antioxidants molecules like anthocyanins that may have therapeutical potential. In this study, we investigated whether fruit extracts from Bactris guineensis and Pourouma cecropiifolia suppress the toxic effects of hydrogen peroxide on astrocytes.

**Methods:** Anthocyanins rich extracts [ARE] from Bactris guineensis and Pourouma cecropiifolia fruits were isolated by high-speed countercurrent chromatography (HSCCC) and preparative HPLC, and their chemical structures were elucidated by using spectroscopic methods.

Once the extracts were obtained, astrocytes were cultured in 96 plate-wells and then treated with different doses of Bactris guineensis and Pourouma cecropiifolia extracts to evaluate their toxicity. Afterwards, to evaluate the protective effect of ARE, astrocytes were pre-treated with non-toxic doses of extracts for 24 hours and then were exposed to hydrogen peroxide for 1 hour. Protective effect was assessed by evaluating changes in cell survival, mitochondrial membrane potential and free radical production by resazurin test and flow cytometry.

**Results:** Our result showed that Bactris guineensis and Pourouma cecropiifolia extracts were not toxic to any of the concentrations evaluated, exhibited a great protective activity, and rescued astrocytes from oxidative peroxide-induced stress by maintaining the mitochondrial membrane potential.

**Conclusions:** These findings suggest that anthocyanins extracts rich in delphinidin-3-O-β-glucopyranoside, cyanidin-3-O-β- glucopyranoside, and cyanidin-3-O-(600-malonyl) glucopyranoside from Bactris guineensis and Pourouma cecropii-

folia can regulate reactive oxygen species via enhancement of mitochondrial function, protecting astrocytes from cell death.

**Keywords:** Antioxidants, Neurodegenerative diseases, Astrocytes, Bactris guineensis, Pourouma cecropiifolia.

(1)Eliana María, Rodriguez; (2)Eliana María, Rodriguez.

(1)210587; (2)Mg. Nutricionist. Facultad de Ciencias Ecáctas y Tecnología. Universidad Nacional de Tucumán. Argentina.

**Background and objectives:** Inulin is stored in many plant species. Research indicates that inulin would have positive effects on the treatment of diabetes. The objectives of this research were to elaborate a low calorie ice cream added with inulin, to determine its chemical composition, sensorial and acceptability, and to compare the variation of pre and post consumption levels of traditional and modified ice cream.

**Methods:** From an experimental study, an ice cream with 10% inulin and 40% of peach pulp, added with sucralose, was made. Its chemical composition was determined and its sensorial properties and acceptability were evaluated in 50 people between 40 and 60 years old of both sexes residing in Tucumán, through tasting and hedonic scale.

**Results:** The prepared ice cream has a caloric value of 49 Kcal / 204 KJ per serving, carbohydrate 3.8 g, protein 0.9 g, total fat 3.3 g, saturated fat 2.2 g, fiber 1 g, and sodium 24 mg. 82% (41) detected a strong, shiny orange hue. 18% (9) did not observe differences between the two ice creams. 90% (45) detected peach aroma and Chantilly cream. 90% (45) reported feeling sweet taste. 88% (44) said that the modified ice cream was creamier than the average. To 84% (42) he liked the modified ice cream. Of the remaining. Regarding the variation in blood glucose levels, people who consumed the common ice cream had 100% normal blood glucose levels (70-110), with a mean of 67.5 μ. While in the post-iced common ice creams 100% had values above 110 mg with a μ of 142.54. In all cases, the glycemia before the modified ice cream was normal (70-110) with a μ of 66.1 and the post-consumption blood glucose levels were 94% normal with a μ of 101.7 and 6% of Values above normal with a μ of 117.3.

**Conclusions:** According to previous scientific evidence, inulin has positive effects on chronic noncommunicable diseases, and according to the data provided by this research, it does not produce significant variations in post-consumption levels of glycemias, therefore, it is proposed to insert in the Market this ice cream to contribute to healthy eating strategies.

**Keywords:** Modified ice cream, Inulin, Low calories.

**Further collaborators:**

Dra. Ing. María Julia Barrionuevo
# A SYSTEMATIC REVIEW EXAMINING THE RELATIONSHIP BETWEEN ANIMAL SOURCE FOOD INTAKE AND GROWTH IN CHILDREN 6 TO 60 MONTHS IN LOW- AND MIDDLE-INCOME COUNTRIES

(1)Shapiro, Myra; (2)Downs, Shauna; (3)Quelhas, Diana; (4)Kreis, Katharine; (5)Kraemer, Klaus; (6)West, Jr., Keith; (2)Fanzo, Jessica.

(1)Phd Candidate. Center for Human Nutrition. Department of International Health. Johns Hopkins Bloomberg School of Public Health. Baltimore. USA.; (2)Phd. Berman Institute of Bioethics. Johns Hopkins University. Baltimore. USA.; (3)PhD. PATH. Seattle. USA.; (4)MPH. Nutrition Innovation. PATH. Seattle. USA.; (5)Sight and Life Magazine; (6)DrPH. Center for Human Nutrition. Department of International Health. Johns Hopkins Bloomberg School of Public Health. Baltimore. USA.

**Background and objectives:** Adequate nutrition is a critical factor in reducing the burden of growth faltering that affects millions of children in low- and middle-income countries (LMIC). This systematic review aimed to examine the association between animal source food (ASF) intake and length/height outcomes in LMIC children aged 6 to 60 months. Secondary outcomes were weight, head circumference (HC), hemoglobin (Hb) concentration, and anemia prevalence.

**Methods:** Databases searched were CINAHL, Embase, Global Index Medicus, PubMed, and Web of Science. Grey literature was searched in International Food Policy Research Institute publications, New York Academy of Medicine Grey Literature Report, OpenGrey, and Proquest Dissertations. Randomized controlled trials (RCT) and longitudinal and cross-sectional observational studies that quantified ASF consumption by frequency, quantity, or both in children 6 to 60 months in LMIC were included.

**Results:** There were 13,431 titles, 205 abstracts, and 89 full texts reviewed. Fourteen articles were included: 4 RCTs, 4 observational cohorts, and 6 cross-sectional. Three RCTs used a plant-based food (PBF) as the comparator, the results showed no differences in height (n=3) or Hb (n=1), mixed positive and null associations for weight (n=1), and higher HC for PBF (n=1). One RCT with a usual diet control resulted in higher Hb, lower anemia prevalence, and no difference in height or weight. In observational cohorts, ASF intake and height had a positive association (n=1), mixed positive/ null (n=1), and negative (n=1). ASF and HC had a positive association (n=1), mixed positive/null (n=1), and no association (n=1); Hb and anemia were not examined. For cross-sectional studies, ASF and height had positive associations (n=1), mixed positive/ null (n=3), and no association (n=2). ASF and weight had mixed positive/null associations (n=3); HC, Hb, and anemia were not examined. Positive or mixed positive/null associations were found in 6 of 13 (6/13) studies for height, 4/9 for weight, 1/2 for HC, and 1/2 for Hb; 1/2 studies had a negative association for ASF and anemia prevalence.

**Conclusions:** The variation in associations may be explained by the heterogeneous quantification of ASF intake. Our results

suggest that increased access to ASF in LMIC may support reducing the burden of growth faltering in early childhood.

**Keywords:** Children, Growth, Animal-source-food, Meat, Poultry.

# TOTAL ANTIOXIDANT CAPACITY AND ITS DIETARY SOURCES IN DIETS OF STUDENTS WITH DIFFERENT PHYSICAL ACTIVITY LEVEL

(1)Czlapka-Matyasik, Magdalena; (2)Przeorska, Dorota; (3)Van Lieshout, Machteld.

(1)Poznan University of Life Sciences; (2)Msc. Food Technologist. Department of Human Nutrition and Hygiene. Poznan University of Life Sciences. Poland; (3)PhD. Food Technologist Nutrition Consultant. Foodwise Pro B.V. Netherlands.

**Background and objectives:** Introduction: Physical activity, determine dietary patterns. Information about dietary sources of antioxidants in daily diet of students is limited.

**Objectives:** The aim of this study was to compare antioxidant capacity of diets students with different physical activity level and their dietary sources of antioxidants.

**Methods:** Method/Design: The study involved 47 subjects aged 19-25 years. The quantitative FFQ questionnaire administered by reviewer was collected and analyzed. Dietary recalls regarding the consumption of dietary sources of antioxidants, socioeconomic data and physical activity were collected. The antioxidant density of daily diet (Q-ORAC; µmolTE/1000 kcal), the share of food groups in total ORAC was evaluated. Daily physical activity of studies group was evaluated according to subject's declaration.

**Results:** Total daily antioxidant density in studied group was 7 056 µmolTE/1000kcal. Sources of total antioxidant capacity in daily students diet come from fruits (52%), vegetables (22%) and spices (25%). Students who were less active consumed less dietary sources of antioxidants, presented lower levels of diet antioxidant capacity. Dietary sources of antioxidants were fruits (52%), vegetables (22%) and spices 25%).

**Conclusions:** We found correlation between dietary sources of antioxidants and physical activity level. Dietary sources of antioxidants should be included in epidemiologic studies and analysed in aspects of physical activity.

**References:**

1. Zujko ME, Witkowska AM, Waskiewicz A, Piotrowski W, Terlikowska KM. Dietary antioxidant capacity of the patients with cardiovascular disease in a cross-sectional study. Nutr J. 2015;14:13. doi:10.1186/s12937-015-0005-4.

2. Czlapka-Matyasik M, Ast K. Total Antioxidant Capacity and Its Dietary Sources and Seasonal Variability in Diets of Women with Different Physical Activity Levels. Polish J Food Nutr Sci. 2014;64(4):267-276. doi:10.2478/pjfsn-2013-0026.

**Keywords:** Antioxidant diet density, Sources of antioxidants, Students, FFQ.
# EVALUATION OF PHYSICO-CHEMICAL CHARACTERISTICS AND ANTIOXIDANT ACTIVITY OF FLOUR JABOTICABA (MYRCIARIA CAULIFLORA) RESIDUE

(1)Thais Resende Teixeira, Karla; (2)Azevedo dos Santos, Talita;

(3)Pimentel de Abreu, Joel; (1)de Souza Gouveia Moreira, Laís; (4)Lima Torres, Taíssa.

(1)Nutritionist of Antonio Pedro University Hospital / Fluminense Federal University, Niterói/ RJ - Brazil.; (2)Nutritionist graduate of Federal University of the State of Rio de Janeiro, Rio de Janeiro / RJ - Brazil; (3)Master of Science in food science at Federal University of the State of Rio de Janeiro, Rio de Janeiro / RJ - Brazil.; (4)Professor of Federal University of the State of Rio de Janeiro, Applied nutrition department, Rio de Janeiro / RJ - Brazil.

**Background and objectives:** The jaboticaba Myrciaria cauliflora (Mart.) is a native Brazilian fruit with high antioxidant potential, but still has a low commercialization and consumption, because of its high perishability and waste generation. The physico-chemical characterization and analysis of the antioxidant activity of the jaboticaba residue provide subsidies to the development of by-products with healthy and sustainable value, to allow greater accessibility to this fruit. The objective of the present study was to evaluate the physical-chemical characteristics and antioxidant activity of jabuticaba flour residue (Myrciaria cauliflora) to be an alternative in food preparation or enrichment.

**Methods:** The jabuticabas were purchased from the Center of Supply of the City of Rio de Janeiro (CADEG) in November 2014, respecting the seasonality of the fruit. The jaboticaba flour residue was obtained by drying the residue resulting from the jabotica-ba juice formulation. The physical and chemical characterization was performed through pH, fixed mineral residue, lipid extraction, protein analysis, total sugars, fiber determination and total titratable acidity. In vitro antioxidant activity studies were carried out, the antioxidant compounds and phenolic compounds were extracted, the TEAC assay and the FRAP assay. Quantification of phenolic compounds was determined by the Folin-Ciocalteau spectrophotometric method.

**Results:** The flour jaboticaba residue showed a yield of 18%. The moisture (11.29%), ash (3.5%) Acidity (1.8%) and pH (3.5) in accordance with legal standards and the percentage of total sugar (62.43%), lipids (0.6%), protein (12.13%) and fibers (9.35%) were adequate. The total antioxidant activity and phenolic of the flour jaboticaba residue showed: 1419.8 micromole Fe 2+ / g (FRAP) and 2505.53 micromole Trolox / g (ABTS) and 5410,3mg GAE / 100g.

**Conclusions:** These results are expressed over various other fruit flour documented in the literature. Therefore the flour jaboticaba residue is a viable way to provide this fruit in a sustainable manner, and to present itself as an alternative source of antioxidants to be used in the preparation of healthy products.

**Keywords:** Flour, Myrciaria cauliflora, Antioxidant, Phenolic acid.

# THE COMBINATION OF MEAL IMPROVEMENT AND PROTEIN-ENRICHED FOOD FOR INCREASING MUSCLE MASS OF THE ATHLETE

(1)Suzuki, Aiyuu; (2)Masaki, Yuusuke; (2)Matsuzawa, Hiroki; (3)Koyama, Yuuzou; (3)Namba, Hideyuki; (1)Murakami, Yukifumi; (3)Matsumoto, Megumi.

(1)MSc; (2)BSc; (3)PhD.

**Background and objectives:** Most athletes aim to improve performance by increasing muscle mass without increasing body fat mass. In this study, in order to establish the ideal weight-increasing method, we examined a diet improvement method using protein-enriched foods and performed a 3-week intervention test.

**Methods:** The subjects were five male shot-put players and eight soccer players in the university students. The intervention period is 3 weeks, and the subjects daily took Lumilan 200 ml (Kewpie Co., Ltd.) containing 84 kcal of energy, 8 g of protein and 0 g of lipid, and took foods containing 2 g of protein per 1 kg body weight, which were based on the high protein food table. Measurements of body composition and meal survey were conducted for 1 week before and after the intervention, and changes in nutrient and amount of food intake were measured.

**Results:** In shot-put players the body composition did not change, but in soccer plyers the body fat mass significantly decreased. Regarding meal intake, the amounts of lipid and dairy products were significantly increased, and cereals intake was significantly decreased in shot-put players. Meanwhile in soccer plyers, the intake amounts of protein, vitamin C, beans, fish and shellfish, and milk were significantly increased. Compared among both athletes, the increments of muscle, fat and body weight were significantly higher in shot-put players than in soccer plyers. Furthermore, increase in intake of protein and lipid was significantly higher in shot-put players. Both players were unable to achieve intake of 2 g protein per 1 kg body weight, so it was thought to be difficult to voluntarily improve diet habit for college student athletes. In addition, it was suggested that 8 g of protein-enriched foods per day were insufficient to achieve increments in 3 weeks.

**Conclusions:** When comparing features of the competition, the shot-put players have to take higher amount of meal for increments than the soccer players, then shot-put players could not achieve necessary intake amount of protein because of the increase of lipid intake accompanied by increased meal intake.

**Keywords:** Soccer, Increment, Protein, Shot-put, Muscle mass
## COMPLEX LIPID COMPOSITION IN HUMAN MILK SAMPLES FROM DIFFERENT ETHNIC GROUPS

(1)Rowan, Angela; (2)Ma, Kevin; (3)McJarrow, Paul; (4)McGibbon, Alastair; (5)Fong, Bertram.

(1)GM Nutrition, Fonterra Research & Development Centre, Palmerston North, New Zealand; (2)MSc, Research Scientist, Fonterra Research & Development Centre, Private Bag 11029, Palmerston North, New Zealand; (3)PhD, Senior Research Scientist, Fonterra Research & Development Centre, Private Bag 11029, Palmerston North, New Zealand; (4)PhD, Principal Research Scientist, Fonterra Research & Development Centre, Private Bag 11029, Palmerston North, New Zealand; (5)MSc, Senior Research Scientist, Fonterra Research & Development Centre, Private Bag 11029, Palmerston North, New Zealand.

**Background and objectives:** Complex lipids, such as gangliosides and phospholipids, play important roles in intra- and inter-cellular signalling, migration, proliferation, neurological development, and inflammatory and immune responses. These lipids are found in biological fluids, including human milk in the milk fat globule membrane.

Growing evidence shows that complex lipids play an important role in infant development, which has led to studies of human milk composition. Furthermore, it has been suggested that human milk composition may be influenced by diet and ethnicity.

The objective of this study was to use modern techniques to determine gangliosides and phospholipids in human milk throughout lactation from a cross section of women of different cultures and ethnicities including Chinese, Malay and the Middle East.

**Methods:** The human milk samples were analysed for gangliosides and phospholipids using a validated high-performance liquid chromatography-mass spectrometry (HPLC-MS) method to determine the ganglioside and phospholipid classes and concentrations.

**Results:** Human milk consisted of both GM3 and GD3. GD3 was the dominant ganglioside in colostrum and transitional milk, while GM3 was the major ganglioside class in mature milk. Total ganglioside concentration was highest in colostrum and transition milk before decreasing in mature milk. Over the mature milk period, there was a gradual increase in average total ganglioside (GD3 +GM3) concentration from around 14 mg/L to high of around 25 mg/L.

Total phospholipid (SM, PC, PE, PI and PS) concentrations were highest in colostrum, before dropping to lower levels at the start of the mature milk period. However the pattern of changes for total phospholipid concentrations over the lactational period differed between ethnic groups.

**Conclusions:** Using modern HPLC-MS techniques, we measured the ganglioside and phospholipid concentrations in breast milk from a number of different ethnic populations across the lactation periods. There were similarities in average total ganglioside and phospholipid concentrations in the mature milk.

These components may be important for normal growth and development of the infant immune system, visual performance

and cognitive performance. Data from this study may support the development of human milk replacers for use in situations where mothers are unable to breastfeed.

**Keywords:** Human milk, Gangliosides, Phospholipids

**Conflict of Interest Disclosure:** This work was supported by the New Zealand Primary Growth Partnership post-farm gate dairy programme, funded by Fonterra Co-operative Group and the New Zealand Ministry for Primary Industries.

**Further collaborators:**

X Liu, M Hamid, H Radwan

## COMPOSITION OF FATTY ACIDS AND CHARACTERIZATION OF BIOACTIVE COMPOUNDS IN THE TUCUMÃ FRUIT (ASTROCARYUM HUAIMI MART.)

(1)Marques Romeiro Santos, Mirelly; (2)Stéphanie Fernandes, Dayane;

(2)Monteiro Santee, Cynthia; (3)Jordão Cândido, Camila; (4)Fontoura Cavalheiro, Leandro; (5)Fernandes da Silva, Anderson; (6)Aragão do Nascimento, Valter; (7)Aiko Hiane, Priscila; (7)Mendes Ramos Filho, Manoel; (8)Freitas dos Santos, Elisvânia.

(1)Doctorate Student in the Health and Development in the Brazilian Midwestern Region Post Graduate Program. Nutritionist. Federal University of Mato Grosso do Sul. Brazil.;

(2)Nutritional Science Undergraduate Student. Federal University of Mato Grosso do Sul. Brazil.; (3)Pharmacist. Food Processing Technitian. Federal University of Mato Grosso do Sul. Brazil.;

(4)Chemist. Institute of Chemistry. Federal University of Mato Grosso do Sul. Brazil.; (5)MSc. Biologist. Federal University of Mato Grosso do Sul. Brazil.; (6)PhD. Professor. Post Graduate Studies in Health and Development in the Brazilian Midwestern Region.

Federal University of Mato Grosso do Sul. Brazil.; (7)PhD. Professor of Pharmacy. Federal University of Mato Grosso do Sul. Brazil.;

(8)PhD. Professor of Nutritional Science. Federal University of Mato Grosso do Sul. Brazil.

**Background and objectives:** The Brazilian Cerrado has a large variation of fruits with different nutritional and physicochemical properties that have not been studied. The purpose of this study is to analyze the antioxidant properties and quantify the fatty acids of the tucumã fruit (Astrocaryum huaimi Mart.) found in the Brazilian state of Minas Gerais.

**Methods:** Total phenols, tannin, and antioxidant activity were analyzed using the Folin-Ciocalteu, Folin-Denis and the DPPH (1,1-diphenyl-2-picryl hydrazyl) stable free radical assay methods, respectively, in the extracts: aqueous, acetone, and ethanolic. For determining fatty acids from tucumã oil Gas-liquid chromatography Varian (model CP-3800) was used. The data was analyzed using the Statistical Package for the Social Sciences (SPSS) software, version 22.0 and reported as average ± standard deviation. The comparisons were done using analysis of variance (ANOVA), followed by Tukey post hoc analysis. The values were statistically significant when p<0.05.
**Results:** The present study indicated a significant differ-
ence (p<0.0001) in the concentration of bioactive compounds in
the different extracts. The tannins in the acetone extract result-
ed in 90.77 ± 1.13 mg EAT.100g-1, greater than in the ethanol-
ic (63.57 ± 1.52) and aqueous (31.10 ± 0.34) extracts. The phe-
nolic compounds were best extracted in ethanol (101.78 ± 1.88
mg EAG.100g-1) when compared to acetone (87.80 ± 2.48) and
aqueous (62.17 ± 0.96). Among the analyzed extracts, the aqueous
presented the least antioxidant potential with a confidence interval
(CI50) equal to 259.95 ± 0.75, followed by the ethanolic (138.49 ±
0.89) and acetone (115.76 ± 5.67). The nutritional composition of
the tucumã oil presented 29.4% of saturated fatty acids, 12.56% of
polyunsaturated fatty acids and 46.2% of unsaturated fatty acids,
with emphasis on oleic acid.

**Conclusions:** In the analysis of bioactive compounds, the acetone extract followed by the ethanolic had the best antioxidant potential and more phenolic compounds and tannins. The quantification of the fatty acids in the fruit presented more unsaturated fatty acids. Based on the results, the inclusion of tucumã pulp in the diet can prevent chronic diseases, as well as nutritional deficiencies, thus promoting health benefits.

**Keywords:** Cerrado, Fruit, Fatty acids, Antioxidants.

**Further collaborators:**

FUNDECT - Fundação De Apoio Ao Desenvolvimento Do
Ensino, Ciência E Tecnologia Do Estado De Mato Grosso Do Sul.

that its endogenous synthesis diminishes in the process of aging.
It has been proven that the dietary enrichment in PA results in an
increase of PA plasmatic levels.

The aim of this work was to estimate the PA intake of the Span-
ish population, mainly focusing in the comparison between elder-
ly and young adult populations.

**Methods:** PA intake estimation was done taking into account the PA contents collected in the database of our research group and food consumption data from Spanish National dietary survey in adults, elderly and pregnant women (ENALIA 2, 2016).

**Results:** Mean PA intake was close to 170 µmol/day for the Spanish population, slightly lower than that estimated from previous surveys in Spain. In comparison with other countries, PA intake in Spain was lower than that previously reported in the United Kingdom and similar to that reported in Japan. The total estimated PA intake for the Spanish population aged 65-74 years (elderly) was higher than that estimated for young adults (18-39 years).

**Conclusions:** The eating habits of the elderly seem to guarantee a high PA intake, principally explained by a higher consumption of fruits and vegetables. This partially compensates the lower efficacy of the endogenous PA synthesis described in this population group.

**Keywords:** Polyamines, Bioactive compounds, Elderly.

144/2563

ESTIMATION OF THE CURRENT POLYAMINE
INTAKE IN THE SPANISH POPULATION

(¹)Vidal-Carou, M. Carmen; (²)Comas-Basté, Oriol; (³)Sánchez-Pérez, Sònia; (⁴)Hernández-Macias, Salvador; (⁵)Latorre-Moratalla, M. Luz;

(¹)Veciana-Nogués, M. Teresa; (⁶)Rivero Urgell, Montserrat.

(¹)PharmD. PhD. Department of Nutrition, Food Sciences and Gastronomy. University of Barcelona. Spain.; (²)PharmD. MSc. Department of Nutrition, Food Sciences and Gastronomy. University of Barcelona. Spain.; (³)BSc. MSc. Department of Nutrition, Food Sciences and Gastronomy. University of Barcelona. Spain.; (⁴)BSc. Department of Nutrition, Food Sciences and Gastronomy. University of Barcelona. Spain.; (⁵)BSc. PhD. Department of Nutrition, Food Sciences and Gastronomy. University of Barcelona. Spain.; (⁶)PharmD. PhD. Associació Catalana de Ciències de l'Alimentació (ACCA). Spain.

**Background and objectives:** For many years, the interest of the study of polyamines (PA) content in foods has lied in the need to formulate low PA diets as part of the treatment of certain neoplastic processes. Currently, however, more attention has been paid on PA potential beneficial effects, both at intestinal and systemic levels. PA are involved in the maturation of the intestinal epithelium in new-borns and have also been postulated as potential protective agents in cardiovascular diseases and diabetes complications due to their anti-inflammatory and antioxidant properties. Moreover, PA have awakened a renewed interest after realising

144/2579

KIMCHI AND ITS BIOACTIVE COMPOUNDS
IMPROVED COGNITIVE DEFICITS IN MICE IN-
DUCED BY AMYLOID BETA

(¹)Song, Yeong Ok, (²)Woo, Minji; (³)Kim, Mijeong; (⁴)Noh, Jeong Sook.

(¹)Department of Food Science and Nutrition. Kimchi Research Institute. Pusan National University. Busan 46241. Republic of Korea.; (²)MSc. Department of Food Science and Nutrition. Kimchi Research Institute. Pusan National University. Busan 46241. Republic of Korea.; (³)PhD. Department of Food Science and Nutrition. Kimchi Research Institute. Pusan National University. Busan 46241. Republic of Korea.; (⁴)PhD. Department of Food Science and Nutrition, Tongmyong University, Busan 48520, Republic of Korea.

**Background and objectives:** Malfunction of cognition is often noticed in the elderly person suffering the dementia wherein elevation in oxidative stress is recognized. Kimchi, Korean fermented vegetable dish, demonstrated numerous health benefits and bioactive compounds in kimchi with antioxidant activity are claimed for revealing these effects. In this study, effects of kimchi and its bioactive compounds on the improvement of cognitive deficits damaged by Aβ25-35 were examined in mice. For the study, permeability of blood-brain barrier (BBB) of kimchi and bioactive compounds were determined in vitro.

**Methods:** Kimchi was mixed with a brined cabbage and condiments prepared with red pepper, garlic, green onion, and ginger. Kimchi was fermented until pH was reached 4.3±0.1. Brain damage was induced by a single intracerebroventricular injection of
Aβ25-35. Following 14 days, mice were fed chow diet and a concomitant oral administration of kimchi methanol extract (KME, 200 mg/kg/day), 3-(4'-hydroxyl-3',5'-dimethoxyphenyl)propionic acid (HDMPPA, 50 mg/kg/day), quercetin (50 mg/kg/day), ascorbic acid (50 mg/kg/day), or capsaicin (10 mg/kg/day), respectively (n = 7 per group). PBS was injected to the mice as a vehicle. Behavior tests including water-maze test, T-maze test, and novel object recognition test were performed. BBB permeability was evaluated using PAMPA assay, in vitro.

**Results:** Compared with the control group, in the water-maze test, the latency time to reach the platform was shorter in the KME and bioactive compounds group (p<0.05). These groups showed a greater preference for the new route than old route in the T-maze test (p<0.05). In the novel object recognition test, KME and bioactive compounds groups spent more time around the novel object than familiar object (p<0.05). Plasma ROS and peroxynitrite level elevated by Aβ25-35 was decreased by KME or bioactive compounds (p<0.05). KME and other compounds except for ascorbic acid were successfully penetrated BBB. The permeability of BBB was high in capsaicin, quercetin, KME, and HDMPPA, in order.

**Conclusions:** Kimchi exerted a preventive effect against the cognitive deficits induced by Aβ25-35 via suppression of oxidative stress, suggesting that kimchi consumption might have benefits on dementia. Synergic effects of bioactive compounds in kimchi rather than a single compound would increase these effects.

**Keywords:** Amyloid beta protein; Blood-brain barrier; Cognitive deficits; Kimchi; Oxidative stress.

**Further collaborators:**

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Education (2015R1D1A1A01057267).

through lipid peroxidation. Hypercholesterolemia is one known fact to disturb the lipid metabolism in the brain. Kimchi, Korean fermented vegetable dish, has demonstrated antioxidative, lipid-lowering, anti-inflammation, and antiatherogenic activities where quercetin, capsaicin, vitamin, mineral, and lactic acid bacteria are present. In this study, beneficial effects of kimchi consumption on alleviating apoptosis mediated through lipid accumulation, oxidative stress, and inflammation in the brain were examined in HCD-fed mice.

**Methods:** Kimchi methanol extracts (KME) prepared from fermented kimchi (pH 4.3±0.1) was used. Animal groups were mice fed high-cholesterol diet (HCD having an oral administration of KME (200 mg/kg bw/day) or distilled water for 8 weeks. The level of oxidative stress and lipid peroxidation were determined in the brain tissue. Gene expressions were analysed by western blot assay to elucidate the related mechanisms.

**Results:** Compared with the control group, expression of lipogenic enzymes (SREBP-1, -2, ACCα, FAS, and HMGCR) in the kimchi group were downregulated with concomitant upregulation of genes involved in the fatty acid oxidation (PPAR-α, CPT1, and ACOX1) and cholesterol export (CYP7A1) (P < 0.05). Reactive oxygen species and lipid peroxidation levels in the brain were significantly reduced (P < 0.05) in the kimchi groups, in contrast, the glutathione level was increased (P < 0.05). Protein expressions such as SOD, CAT, and GPx regulated by Nrf2 were increased (P < 0.05). Moreover, NFκB and their related inflammatory gene expressions were significantly downregulated (P < 0.05). Additionally, expression for proapoptotic factors (Bax, caspase -3, and -9) was downregulated while those for anti-apoptotic factor (Bcl-2) were upregulated (P < 0.05).

**Conclusions:** Kimchi decreased apoptosis in the brain of mice fed HCD through decreasing lipid accumulation, inflammation, and oxidative stress, suggesting that kimchi consumption might improve the brain function.

**Keywords:** Brain; High-cholesterol diet; Kimchi; apoptosis; Oxidative stress; Inflammation.

**Further collaborators:**

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Education (2015R1D1A1A01057267).

<footnote>(1)Song, Yeong Ok; (2)Kim, Serim; (3)Woo, Minji; (4)Kim, Mijeong; (5)Noh, Jeong Sook.</footnote>

<footnote>(1)Department of Food Science and Nutrition. Kimchi Research Institute. Pusan National University. Busan 46241. Republic of Korea.; (2)BSc. Department of Food Science and Nutrition. Kimchi Research Institute. Pusan National University. Busan 46241. Republic of Korea.; (3)MSc. Department of Food Science and Nutrition. Kimchi Research Institute. Pusan National University. Busan 46241. Republic of Korea.; (4)PhD. Department of Food Science and Nutrition. Kimchi Research Institute. Pusan National University. Busan 46241. Republic of Korea.; (5)PhD. Department of Food Science and Nutrition, Tongmyong University, Busan 48520, Republic of Korea.</footnote>
# BENEFICIAL EFFECTS OF RICE ENDOSPERM PROTEIN ON CHRONIC KIDNEY DISEASE-MINERAL AND BONE DISORDER (CKD-MBD) IN ZUCKER DIABETIC FATTY RATS

**Background and objectives:** The increase of chronic kidney disease (CKD) is a serious problem worldwide. In Japan, CKD patients are estimated 13.3 million, about 13% of adults, and diabetic patients are 9.5 million. Diabetes and CKD are directly related, and CKD including diabetic nephropathy leads to CKD-mineral and bone disorder (CKD-MBD). Moreover, CKD-MBD is one of the major risk factors for cardiovascular disease as well as bone fractures. We have reported that rice endosperm protein (REP) had preventive effects on obese diabetes, fatty liver and diabetic nephropathy, compared with casein (C). This study is the advanced experiment and the aim is to clarify the effects of REP on the bone metabolism in Zucker Diabetic Fatty (ZDF) rats.

**Methods:** Seven week old male ZDF rats were fed 20% C and REP diets for 10 weeks. Fasting blood glucose levels and urinary albumin excretion (UAE) were monitored. At the end of the experimental period, some blood parameters related to the mineral metabolism (Ca, P, parathyroid hormone (PTH), and fibroblast growth factor-23 (FGF-23)) and a blood parameter related to the bone formation (osteocalcin) were determined to evaluate the progression of CKD-MBD. The femur bones were used for histomorphometric observation by µCT, and the bone strength was measured. In addition, the urinary metabolome measurements were conducted.

**Results:** Fasting blood glucose levels and UAE in the REP group were significantly suppressed compared with those in the C group. In the REP group, blood FGF-23 concentrations was significantly decreased although blood Ca and P concentration had no abnormal alteration in ZDF rats. Moreover, the osteocalcin level significantly increased in the REP group. The REP had no significant effects on bone mineral density but suppressed deterioration of bone microstructure parameters, and the bone strength was improved in the REP group. PCA analysis showed clearly different urinary metabolic profiles in the two groups.

**Conclusions:** Dietary REP delays the progression of CKD-MBD and improves the bone strength, compared with C in ZDF rats.

**Keywords:** Rice endosperm protein, CKD-MBD, Diabetes, ZDF rat

# BENEFICIAL EFFECT OF RICE ENDOSPERM PROTEIN ON IGE PRODUCTION

**Background and objectives:** Rice is a staple food and an important food not only as an energy source but also as a protein source especially in Asian countries. There are some unknown areas as for the physiological functions, especially immune system, of rice endosperm protein (REP) although the effect of formulae based on rice protein has been reported in the treatment with milk protein allergy. In our preliminary study, REP affected intestinal gene expressions related to immune system and also REP enhanced cellular immunity and deflected the balance of type I helper T cell (Th1)/type II helper T cell (Th2) to Th1 side in Lewis rats. Therefore, we attempted to clarify the effect of REP on the Th1/Th2 balance and IgE production.

**Methods:** Female BALB/c mice at 3 weeks of age were employed and were fed on an egg white protein (EWP) diet as a control group and an REP diet for 4 weeks. To assess the Th1/Th2 balance and IgE production, blood samples were collected from tail vein at 4 weeks, and IgG2a, a marker for Th1 type immune response, IgG1, a marker for Th2 type immune response, and IgE concentrations were measured by the ELISA method.

**Results:** At 4 weeks, the serum IgG1 concentration was significantly suppressed and IgG2a/IgG1 ratio was significantly increased by REP consumption although IgG2a concentration was not altered between two groups. This result suggests that REP deflected the Th1/Th2 balance to Th1 side. In addition, the serum IgE concentration in the REP group was significantly suppressed at 4 weeks compared with the EWP group.

**Conclusions:** REP deflected the balance of Th1/Th2 to Th1 side and suppressed the IgE production compared with EWP.

**Keywords:** Rice endosperm protein, Th1/Th2 balance, IgE

<footnote>(1)Watanabe, Reiko; (2)Kubota, Masatoshi; (3)Sugaki, Shogo;</footnote>

<footnote>(4)Hosojima, Michihiro; (5)Saito, Akihiko; (6)Fujii, Mikio; (7)Sasagawa, Ichiro; (8)Fujimura, Shinobu; (9)Kadowaki, Motoni.</footnote>

<footnote>(1)PhD. Professor. University of Niigata Prefecture; (2)PhD. Lecturer. Niigata University of Pharmacy and Applied Life Sciences. Japan.; (3)MSc. Student. Graduate School of Science and Technology. Niigata University. Japan.; (4)MD. PhD. Associate Professor. Graduate School of Medical and Dental Sciences. Niigata University. Japan.; (5)MD. PhD. Professor. Graduate School of Medical and Dental Sciences. Niigata University. Japan.;</footnote>

<footnote>(6)PhD. Senior Researcher. Kameda Seika Co. Ltd. Japan.; (7)PhD. Professor. School of Life Dentistry at Niigata. The Nippon Dental University. Japan.; (8)PhD. Associate Professor. Graduate School of Science and Technology. Niigata University. Japan.; (9)PhD. Professor. Graduate School of Science and Technology. Niigata University. Japan.</footnote>

<footnote>(1)Kubota, Masatoshi; (2)Kamada, Takuya; (3)Hara, Takashi; (4)Higuchi, Yuki; (5)Fujii, Mikio; (6)Sato, Shinji; (3)Fujimura, Shinobu; (3)Kadowaki, Motoni.</footnote>

<footnote>(1)PhD. Faculty of Applied Life Sciences. Niigata University of Pharmacy and Applied Life Science. Japan.; (2)MSc. Graduate School of Science and Technology. Niigata University. Japan.;</footnote>

<footnote>(3)PhD. Graduate School of Science and Technology. Niigata University. Japan.; (4)MSc. Kameda Seika Co. Ltd. Japan.; (5)PhD. Kameda Seika Co. Ltd. Japan.; (6)PhD. Faculty of Applied Life Science. Niigata University of Pharmacy and Applied Life Science. Japan.</footnote>
## COLITIS IMPROVEMENT EFFECT OF PYRO-GLUTAMYL PEPTIDES-RICH FERMENTED RICE DRINK IN MICE

(1)Wada, Sayori; (2)Kasuno, Toko; (3)Kiyono, Tamami; (2)Aoi, Wataru;

(4)Naito, Yuji; (5)Sato, Kenji; (2)Higashi, Akane.

(1)Kyoto Prefectural University; (2)Division of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Kyoto, Japan; (3)Kyoto Municipal Institute of Industrial Technology and Culture, Kyoto, Japan; (4)Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine, Kyoto, Japan; (5)Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.

**Background and objectives:** Three pyroglutamyl peptides with anti-colitic activity had been identified in Japanese rice wine, sake, by oral administration of a small dose (0.1-1.0 mg/kg body weight) in mice. Since the alcohol percentage of sake is approximately 15%, we developed a functional peptides-rich fermented rice drink with no alcohol, for human application. We also compared the anti-colitic activity of developed fermented rice drink with Japanese traditional drinks, including sake and amazake, a traditional sweet, low- or non-alcohol Japanese drink.

**Methods:** For preparation of fermented rice drink, steamed rice, koji, lactic acid, and water were fermented according as amazake, but at low temperter for 1 week. Acute colitis was induced by oral administration of dextran sulphate sodium dissolved in drinking water for 7 days in mice. 4 samples (fermented rice drink, amazake, 2 different types of sake) were dissolved in distilled water and administered by gavage in aliquots of 200 microL once a day during the entire induction. The mice were sacrificed on day 7 the entire colon was removed from the cecum to the anus. We evaluated the colitis severity on the basis of microscopic observations of stool consistency (0, normal; 1, mild loose; 2, loose; 3, mild diarrhea; and 4, diarrhea). The inner contents of the colon were collected for microbiota analysis. Population of Bacteroiditis and Firmicutes were quantified on the basis of amplification of genome DNA coding 16S rRNA by polymerase chain reaction by using group-specific primers.

**Results:** Fermented rice drink group had a tendency to improve diarrhea score (1.6 ± 0.2, p = 0.061) compared with colitis group (2.7 ± 0.4), while Japanese traditional drink (amazake and sake) did not improve. There were no significant difference in the population of Firmicutes/Bacteroidetes ratio between colitis and rice drink group.

**Conclusions:** We developed novel no-alcohol functional rice drink, and it suppressed diarrhea to a greater extent than did the Japanese traditional drinks in mice. Fermented rice drink might hold promise for human application.

**Keywords:** Pyroglutamyl peptides, Fermented foods, Gut microbiota, Colitis.

## EFFECTS OF BIOACCESSIBLE GARLIC COMPOUNDS ON CULTURED VASCULAR SMOOTH MUSCLE CELLS FROM SPONTANEOUSLY HYPERTENSIVE RATS

(1)Torres, Carolina; (2)Redondo, Analía; (3)Quesada, Isabel; (4)Camargo, Alejandra; (3)Castro, Claudia.

(1)MSc. Food Science. Instituto de Biología Agrícola - CONICET - Universidad Nacional de Cuyo. Argentina.; (2)MSc. Biologist. Instituto de Medicina y Biología Experimental de Cuyo - CONICET - Universidad Nacional de Cuyo. Argentina.; (3)PhD. Instituto de Medicina y Biología Experimental de Cuyo - CONICET - Universidad Nacional de Cuyo. Argentina.; (4)PhD. Instituto de Biología Agrícola - CONICET - Universidad Nacional de Cuyo. Argentina.

**Background and objectives:** Several studies have associated a moderate consumption of garlic with improvements in health. The evidence suggests that its medicinal activities are mainly due to the organosulfur compounds (OSCs) garlic content. Garlic compounds have exhibited antioxidant activities and cardiovascular disease-protective properties. The remodeling of small arteries by excessive proliferation and migration of vascular smooth muscle cells (SMCs) is a phenomenon to consider, since it contributes to the development and complications of hypertension, one leading cause of morbidity and mortality globally. It seems that the mystery of OSCs and hypertension would be easy to elucidate; nevertheless, OSCs have shown to have different biological actions, potency and mechanisms in different cell lines and organ tissues. Moreover, only the bioavailable compounds would have bioefficacy. In this regard, previous works have identified a few OSCs in blood. For this study three bioaccessible OSCs, allyl mercaptan (AM), S-allyl cysteine (SAC) and 2-vinyl-4H-1,3-dithiin (2VD), were selected to evaluate their in-vitro effects on cultured SMCs isolated from mesenteric arteries of male spontaneously hypertensive rats.

**Methods:** Cell proliferation, cell migration and intracellular oxidative stress were evaluated after cell exposure to OSCs expected quantities in plasma, 10 µg/L of AM and 2VD and 50 µg/L of SAC.

**Results:** The results of the proliferation test by MTT assay showed that 2VD reduced by 20% the proliferation compared to the control, conversely SAC enhanced the proliferation by 15%. Scrape-wound migration assay showed that 2VD and AM reduced the cell migration in similar ratios (30%), whereas SAC increased the migration by 30% compared to the control. Concerning to the intracellular ROS levels, the compounds showed different behaviors at the studied concentrations. While AM was an intracellular antioxidant, 2VD and SAC increased the oxidative stress.

**Conclusions:** In conclusion, expected quantities of 2VD and AM in plasma, provided by garlic intake, were able to inhibit the migration of SMCs of spontaneously hypertensive rats. These compounds could help to protect the cardiovascular system from vascular remodeling produced by hypertension. Further studies will be needed to elucidate the specific mechanism of action to
inhibit the SMCs migration and the role of its different antioxidant effects.

**Keywords:** Organosulfur compounds, Garlic, Hypertension, Vascular remodeling.

## UTILITY OF DEWAXED BROWN RICE AS A FOOD STAPLE IN THE MAINTENANCE AND IMPROVEMENT OF NUTRITION AND HEALTH STATUS IN THE ELDERLY

**Background and objectives:** A new type of brown rice with its wax layer removed (dewaxed brown rice) resulting quick-cooking and tasty product contains 100-times more LPS than polished white rice. It also improves weight loss and constipation in antibiotics administered mouse model. In this study, the health maintenance function of dewaxed brown rice was investigated its effects on physical and nutritional status, bowel movements, immune function, cognitive function and other factors in residents of special nursing homes for the elderly who continuously ingested dewaxed brown rice.

**Methods:** This study was a controlled crossover trial in 35 elderly men and women living in special nursing homes for the elderly who were sorted into two groups and ingested either dewaxed brown rice or polished white rice three times a day for six months. The subjects underwent body weight measurements once a month, were checked daily for any bowel movements, and underwent salivary sIgA and C-reactive protein (CRP) measurements once every three months and assessments of cognitive function (Hasegawa dementia scale) once every six months. (Ethics Committee of Nagoya University of Economics: receipt number 2016-1).

**Results:** An interim assessment was conducted six months after the study began. In terms of physical status, body weight and body mass index (BMI) tended to increase in emaciated subjects and were significantly greater (p<0.05) after three months of ingesting the test food compared with before the study began. The mean number of days per month without a bowel movement tended to decrease in the dewaxed brown rice group. Salivary sIgA levels in the dewaxed brown rice group showed an increasing trend compared with before the study began, while CRP levels showed a decreasing trend. Assessments of cognitive function in the de-

waxed brown rice group showed improvement compared with those in the polished white rice group.

**Conclusions:** The results of this study suggested that dewaxed brown rice improves constipation, weight loss and inflammation in the elderly, and if introduced as a food staple to residents of welfare facilities for the elderly, could help improve constipation and malnutrition and prevent infections and cognitive decline in the elderly.

**Keywords:** Dewaxed brown rice. Nutritional status. Constipation. Immune system. Elderly.

**Conflict of Interest Disclosure:** Nagoya University of Economics. Scholarship donation: Toyo Rice Corp.. Wakayama Japan

## PURIFICATION OF BIOACTIVE ANTIOXIDANT AND ANTIHYPERTENSIVE QUINOA (CHENOPODIUM QUINOA WILLD.) PEPTIDES

**Background and objectives:** Quinoa is a pseudo Andean cereal known for its protein quality. Also, there are studies that had shown antioxidant and antihypertensive capacity of quinoa peptides. Thus, the aim of this work was to purify bioactive peptides from quinoa protein concentrate through ultrafiltration (10 and 3 kDa cut off) and low pressure chromatographic techniques (adsorption and gel filtration chromatography), in order to improve their antioxidant and antihypertensive capacity.

**Methods:** -Determination of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) antioxidant capacity (umol Trolox equivalent/g protein)

-Determination of antihypertensive capacity through concentration of quinoa peptide to inhibit the angiotensin converting enzyme (ACE) by fifty percent (IC50)

-Adsorption chromatography with ethanol as eluent in steps of 0, 25, 50 and 75% aqueous solution of ethanol

-Gel filtration chromatography with Milli-Q water as isocratic eluent

**Results:** -It was found an antioxidant capacity of 1,667.38±9.55 µmol trolox equivalent (TE)/g protein and 0.3007 mg/mL of IC50 value in quinoa hydrolyzate.

-The fraction obtained from a 3 kDa ultrafiltration cut off got higher both, antioxidant and antihypertensive capacity than the quinoa hydrolyzate.

-When both (adsorption and gel filtration chromatography, in that order) separation techniques were used after 3 kDa ultrafiltration cut off, the highest antioxidant (3,784.85±17.06 µmol TE/g protein) and antihypertensive capacity (IC50 = 0.0391±0.01

<footnote>(1)Uenobe, Maya; (2)Wakuaku, Natsumi; (3)Saika, Toshiyuki;</footnote>

<footnote>(3)Nishiyama, Naoki; (4)Itou, Kazuhiro; (5)Inagawa, Hiroyuki.</footnote>

<footnote>(1)PhD. MD. Department of Health and Nutritional Science. Nagoya University of Economics. Aich Japan; (2)Course of Nutrition Management. Graduate School of Human Life Science. Nagoya University of Economics. Aich Japan; (3)Toyo Rice Corp.. Wakayama Japan; (4)Heiann no Sato. Special Nursing Home for the Elderly. Aich Japan; (5)PhD. Phar.D.. Department of Research & Development. Control of Innate Immunity Technology Research Association, Kagawa Japan.</footnote>

<footnote>(1)Cisneros, Miluska; (2)Campos, David; (3)Aguilar, Ana María.</footnote>

<footnote>(1)BSc. Food Engineering. Industrial Biotechnology Institute. National Agrarian University La Molina. Peru; (2)Dr. Agronomic Sciences. Industrial Biotechnology Institute. National Agrarian University La Molina. Peru; (3)Dra. Agronomic Sciences. Industrial Biotechnology Institute. National Agrarian University La Molina. Peru.</footnote>
mg/mL) were obtained, in comparison to only apply gel filtration chromatography.

-It was found a 82.05 percent of correlation between antioxidant and antihypertensive capacity, showing that both functional properties are related to each other

**Conclusions:** The fraction that presented the highest antioxidant and antihypertensive capacity in the first purification stage, corresponded to the permeate obtained with the cut off membrane 3 kDa

-In the adsorption chromatography, the highest antioxidant and antihypertensive capacity were attributed to the peptides eluted with a 25% aqueous solution of ethanol, indicating the slight hydrophobicity of said peptide sequence

-During the gel filtration chromatography, better results were obtained in peptides previously subjected to adsorption chromatography, which represent those with lower molecular weight and hydrophobic character

-Purification achieved the increase of both, antioxidant and antihypertensive capacity in bioactive quinoa peptides, with a high correlation between each other

**Keywords:** Quinoa, Antioxidant peptides, Antihypertensive peptides, Purification.

**Further collaborators:** Adelayda Pardo

144/2685

A HEALTHY POSSIBILITY: MALTED FLOUR FROM RYE GROWN IN ARGENTINA

(1)Zuleta, Ángela; (2)Zuleta, Ángela; (2)Moreyra, Federico; (2)García, Pilar Teresa; (2)Gómez Castro, Maria Laura; (2)Larregain, Claudia Cecilia; (2)Coscarello, Ethel Noemi; (2)Weisstaub, Adriana Ruth.

(1)university of buenos aires; (2)BUENOS AIRES.

**Background and objectives:** Rye is the second most important winter forage cereal in Argentina. The flour made from rye has a greater proportion of dietay fiber than wheat flour.. Dietary fiber (DF) has beneficial physiological effects on the human organism. Increased intake of DF through foods is actually being recommended for good health because DF is likely to help reduce diabetes and intestinal diseases. Some components of soluble fiber called prebiotics, like beta glucans or aranbioxilans, are defined as non-digestible food ingredients that positively affect the host. The quality of DF varies according to the type of cereals and to the process that is carried out to obtain the flour. Rye grains for this work came from the INTA, Bordenave Púan. The production of flours, with functional properties implies the malting of the rye, and there are antecedents by which the malting process increases the amount of soluble fibers.

**Methods:** In the first instance, an evaluation of the quality of the seeds of the different varieties was carried out. The most efficient rye varietes after the malting process are Don Ewald and Camilo. In order to determinate their characteristics, we analyzed the composition of these varieties . The total, soluble and insoluble fiber content was determined by Method 991.43 AOAC, with flours obtained from the unmalted and malted cereal.

**Results:** The comparative results demonstrate a tendency to modify the amount of fiber in cereal flour after the malting process. Our results show a decrease in the insoluble fiber fraction and an increase in the soluble fiber fraction. The total fiber content of malted and unmalted rye flour was on average 18%. The insoluble fiber content of unmalted rye flour was 16% and malted rye flour was 14%, obtaining of soluble fiber content of 2% in the unmalted flour and 4% in the malted flour. This functional property of bioactive ingredients is currently being evaluated in an animal model of Wistar rats.

**Conclusions:** Preliminary results are showing a functional activity of these ingredient.

**Keywords:** Rye flour, Malted rye, Dietary fiber.

144/2743

SYNERGISTIC EFFECT OF WALNUT AND BLACK CHOKEBERRY MIXTURE ON ANTI-LIPID PER-OXIDATION IN D-GALACTOSE INDUCED AGING MODEL

(1)Song, Eunkyung; (2)Choi, Yoon jeong; (2)song, Mijeong; (3)Kim, Hyun-Sook.

(1)MSc. Student. Food and Nutrition. Sookmyung Women's University. Seoul. Korea.; (2)BSc. Student. Food and Nutrition. Sookmyung Women's University. Seoul. Korea; (3)PhD. Professor. Food and Nutrition. Sookmyung Women's University. Seoul. Korea.

**Background and objectives:** Many studies mentioned the cellular damage caused by reactive oxygen species (ROS) could be aging. D-galactose causes aging-related changes and oxidative stress in the organism. Lipids are the first target of free radicals, and lipid peroxidation related the aging process. Walnut kernels (Juglans regia L.) contains antioxidant phenolic compounds. Many studies showed that walnut phenolics belong to ellagitannin, or hydrolyzable tannins related antioxidant activities. Chokeberry(Aronia melanocarpa) is one of the richest sources of polyphenols, especially anthocyanin among other fruits. Previous studies showed that polyphenols of chokeberry have antioxidant and anti-inflammatory activities. In this study, we mainly investigated that synergistic effect of walnut and chokeberry mixture on anti-lipid peroxidation of the serum and various tissues in D-galactose induced aging model.

**Methods:** Thirty-five Balb/c mice (8 weeks of age) were divided into five groups (n=7 for each group): the normal control group (NC), D-galactose control group (DC), D-galactose with chokeberry diet group (CHD), D-galactose with walnut diet (WD), and D-galactose with walnut and chokeberry mixture diet (WCH). All groups were intraperitoneally infected with D-galactose (120mg/kg) daily for eight weeks, except for NC group which was treated with saline. Levels of malondialdehyde (MDA) in serum, liver, brain, and kidney were determined by according assay kits in D-galactose treated mice.
**Results:** It was found that the serum, hepatic, and brain MDA level in DC groups were higher than the NC group(p<0.05). Compared to DC group, CHD, WD and WCH groups showed significantly decrease serum, hepatic, and kidney MDA level(p<0.05). Serum MDA level in WD group was the lowest concentration compared to other groups. Hepatic MDA level in CH group was the lowest concentration compared to other groups, and also hepatic MDA level in WCH group lower than WD group. There was the lowest MDA level in kidney than WD and CHD groups.

**Conclusions:** Walnut and chokeberry mixture had anti-lipid peroxidation and synergistic effect. Especially, walnut and chokeberry mixture had been decreased the lipid peroxidation product in kidney compared walnut and chokeberry, respectively. But results showed that walnut and chokeberry mixture had not anti-lipid peroxidation in serum, liver, and brain. Further studies are needed to the mechanism of the lipid peroxidation and synergistic effect of various tissues.

**Keywords:** Synergistic effect, Walnut, Chokeberry, Lipid peroxidation, Aging

144/2750

EVALUATION OF THE ANTI-INFLAMMATORY/
ANTIOXIDANT EFFECT OF AQUEOUS LEAVES
EXTRACT OF PASSIFLORA ALATA CURTIS AND
CATECHIN, RUTIN, ISOORIENTIN, VITEXIN IN
CO-CULTURE OF MIN6/LYMPHOCYTES FROM
NOD MICE

(1)Silva, Priscila Matos; (2)Silva, Priscila Matos; (3)Figueiredo, Daniella;

(3)Colomeu, Talita Cristina; (3)Schumacher, Nayara Simon Gonzales;

(4)Carvalho, Virgínia de Campos; (2)Abram, Débora Moitinho;

(5)Fernandes, Luís Gustavo Romani; (6)Zollner, Ricardo Lima.

(1)MSc. Student. Biomedical.n Laboratory of Translational Immunology. Department of Internal Medicine. University of Campinas. São Paulo. Brazil; (2)MSc. Student. Biomedical. Laboratory of Translational Immunology. Department of Internal Medicine. University of Campinas. São Paulo. Brazil; (3)PhD. Student. Biologist. Laboratory of Translational Immunology. Department of Internal Medicine. University of Campinas. São Paulo. Brazil; (4)MSc. Student. Biologist. Laboratory of Translational Immunology. Department of Internal Medicine. University of Campinas. São Paulo. Brazil; (5)PhD. Biologist. Laboratory of Translational Immunology. Department of Internal Medicine. University of Campinas. São Paulo. Brazil; (6)PhD. Doctor. Laboratory of Translational Immunology. Department of Internal Medicine. University of Campinas. São Paulo. Brazil.

**Background and objectives:** Type 1 Diabetes leads to the increase of reactive oxygen species, promoting oxidative stress in the organism. This study aimed to evaluate the effects of the treatments with aqueous leaves extract of P. alata or isolated compounds catechin, rutin, isoorientin and vitexin in the inhibition of proliferation of TCD4+ and TCD8+ cells of NOD mice in co-cul-

ture with MIN6 cells and also the antioxidant effects in the supernatants of these cultures.

**Methods:** T cells (2,5x10⁵) were submitted to co-culture with MIN6 cells (8x10⁴) to determine the concentrations of aqueous extract of P. alata and isolated compounds in inhibition of T cell proliferation. After that, the supernatants were collected for antioxidant activity analyses by FRAP and ABTS protocols and oxidative stress by TBARS protocol.

**Results:** The concentration that promoted inhibition in the cell proliferation in approximately 50% (ID50) was 500ug/mL of P.alata; 50uM of catechin; 300uM of rutin; 5uM of isoorientin and 200uM of vitexin. The FRAP analysis showed higher antioxidant activity in the treatment with rutin, catechin and P. alata while in the ABTS analysis it was verified higher antioxidant activity in the treatment with rutin and vitexin. These both techniques (FRAP and ABTS) demonstrated an increase in capacity to reduce preforms oxidized ions in the groups treated with P. alata, rutin, catechin and vitexin. This activity is associated to antioxidant and anti-inflammatory action, enhancing the protective action in experimental model diabetes type 1. In TBARS analyses, the compound rutin showed a tendency in decrease lipid peroxidation levels when compared to cells that were not submitted to treatment.

**Conclusions:** All the treatments presented inhibition in T cells proliferation, suggesting an anti-inflammatory effect and P. alata and the isolated compounds rutin, catechin and vitexin have showed an antioxidant effect when analyzed the supernatants. The oxidative stress verified by TBARS suggest that rutin is the main isolated compound that decrease the lipid peroxidation. These antioxidant properties from P. alata and compounds can indicate a protection against cell damage from oxidative stress observed in type 1 diabetes evolution.

**Keywords:** Oxidative stress, Lipid peroxidation, Passiflora alata Curtis, Anti-inflammatory effect; NOD mice.

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

144/2781

NEUROPROTECTIVE EFFECT OF SALVIA HIS-
PANICA I (CHIA) OIL AGAINST PESTICIDES IN
RATS

(1)Oriondo Gates, Rosa Lorenza; (2)Valdivieso Izquierdo, Lazaro Ruben; (2)Delgado Pérez, Doris.

(1)Docente miembro investigador Instituto Centro de Investigaciones de Bioquímica y Nutrición. Peru; (2)Docente miembro investigador Instituto Centro de Investigaciones de Bioquímica y Nutrición"Alberto Guzmán Barrón" Facultad de Medicina - UNMSM.Peru

**Background and objectives:** Salvia hispanica (chia) oil has considerable amounts of polyunsaturated fatty acids (PUFAs), similar to linseed oil. The oil could be used as a potential source of high functional ingredients in omega-3 fatty acids. On the other hand toxicological studies on exposure to neurotoxic substances in humans have shown that the central nervous system (CNS) is
higher levels than the group DHCB. In addition, the DHC group presented a higher glucose intolerance compared with DHCB and control. The fecal lipid excretion was higher in group DHCB.

**Conclusions:** The biomass of green bananas to a high calorie diet supplemented gives a protection to changes caused by this diet on lipid profile and glucose Swiss mice, probably by increasing the fecal excretion of lipids.

**Keywords:** Green Banana, Resistant starch, Swiss mice, Hypercaloric diet.

**Conflict of Interest Disclosure:** The authors certify that they have NO affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers' bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this abstract

**Further collaborators:** This study was supported by Scientific Program of Redentor University

144/2796

## INTAKE OF RED WINE GRAPE POMACE FLOUR INCREASES ANTIOXIDANT FUNCTION OF HIGH DENSITY LIPOPROTEINS (HDL) IN HUMANS

**Background and objectives:** In recent years, new antiatherogenic mechanisms have been attributed to high density lipoproteins (HDL), including their ability to prevent lipid peroxidation, a key pathogenic process underlying a wide variety of chronic diseases. Natural antioxidant components present in certain foods are able to prevent this oxidative process. Thus, the food industry is leading a major effort to develop and increase consumption of

antioxidant-enriched products for improving health and preventing disease in the general population. In Chile, approximately 10 mill. hectoliters of wine are produced per year, a process by which 20% of the total grape weight is lost as pomace waste. Red wine grape pomace (RWGP), a by-product derived from winemaking, is composed mainly of grape skin and seeds. This pomace is a valuable resource for production of fiber and antioxidant-rich flour that can be used as ingredient in the development of functional foods.

To evaluate the effect of RWGP flour consumption on the antioxidant capacity of HDL in humans.

**Methods:** While one group of subjects added 20 g RWGP flour/day to their usual diet for 4 months (treated group, n=12), another group consumed their usual diet during the same time (control group, n=12). Blood samples were taken from all volunteers, before and after the intervention. Plasma was used to evaluate total cholesterol content and lipoprotein cholesterol profile. In addition, antioxidant capacity of total plasma and gel filtration-purified HDL particles was determined through an in vitro fluorescence-based assay.

**Results:** Compared with controls, subjects who consumed a diet supplemented with RWGP flour during 4 months showed an increase in the antioxidant capacity of total plasma and purified HDL, without significant changes in total cholesterol or lipoprotein cholesterol levels.

**Conclusions:** Intake of RWGP flour -presumably due to its content of biologically active components- exhibits a favorable effect in vivo by improving the antioxidative functionality of HDL in humans. Thus, widespread use of this functional ingredient may be beneficial in the prevention and treatment of atherosclerotic cardiovascular disease.

This work was supported by grants FONDECYT #1150399 and FONDEF #AF10i1014.

**Keywords:** Wine grape pomace flour. HDL. Antioxidant function.

144/2810

## EFFECT OF THE CONSUMPTION OF MORINGA OLEIFERA IN DIABETIC RATS

**Background and objectives:** The use of Moringa oleifera (Mo) has been evaluated in diseases such as Diabetes mellitus for its possible ability to lower glucose concentration as it contains

<footnote>(1)Rivera, Katherine; (2)Mendoza, Camila; (3)Echeverría, Guadalupe;</footnote>

<footnote>(4)Urquiaga, Ines; (5)Amigo, Ludwig; (6)Busso, Dolores; (7)Rigotti, Attilio.</footnote>

<footnote>(1)Biochemical. Departamento de Nutrición, Diabetes y Metabolismo. Facultad de Medicina. Pontificia Universidad Católica de Chile. Chile.; (2)Biochemist, M. Sc. in Nutrition. Departamento de Nutrición, Diabetes y Metabolismo. Facultad de Medicina. Pontificia Universidad Católica. Chile.; (3)Engineer, M. Sc. in Bioprocessing. Centro de Nutrición Molecular y Enfermedades Crónicas. Facultad de Medicina. Pontificia Universidad Católica. Chile.; (4)Biochemist, Ph.D. Centro de Nutrición Molecular y Enfermedades Crónicas. Facultad de Medicina. Pontificia Universidad Católica. Chile.; (5)Biochemist. Departamento de Nutrición, Diabetes y Metabolismo. Facultad de Medicina. Pontificia Universidad Católica. Chile.; (6)Biologist, Ph.D. Departamento de Nutrición, Diabetes y Metabolismo. Facultad de Medicina. Pontificia Universidad Católica. Chile.; (7)M.D., Ph.D. Departamento de Nutrición, Diabetes y Metabolismo and Centro de Nutrición Molecular y Enfermedades Crónicas. Facultad de Medicina. Pontificia Universidad Católica. Chile.</footnote>

<footnote>(1)Piña Cruz, Rubén; (2)Villarruel-López, Angélica; (2)Vázquez-Paulino, Olga Deli; (3)Guerrero-Quiroz, Luis Alfonso; (4)López-Dela Mora, David Alejandro; (4)Nuño-Anguiano, Karla Janette.</footnote>

<footnote>(1)Student of Nutrition Degree. University Center of Tonala. Guadalajara University. Mexico.; (2)PhD. Department of Pharmacobiology. University Center of Exact Sciences and Engineering. University of Guadalajara. Mexico.; (3)PhD. University Center of Agricultural Biological Sciences. University of Guadalajara. Mexico.; (4)PhD. Department of Health Sciences. University Center of Tonala. University of Guadalajara. México.</footnote>
polyphenols (quercetin, kaempferol, glucosides). However, most studies have been done with the extract of the plant, which does not reflect the actual way of consumption as the population consumes it in powder.

Evaluate Mo powder to study the effects on glucose, triglycerides, Cholesterol, body weight, toxicity, genotoxicity and predominance of microbiota in diabetic rats.

**Methods:** Four groups were established for the toxicity test: control and Mo with doses of 100, 200 and 500 mg/kg (Milligrams per kilogram of weight) respectively, which were used for the micronucleus count in erythrocytes. Another 5 groups were used for the diabetic model. In addition, glucose, body weight, triglycerides, cholesterol, HDL and LDL were measured. Histological sections of the liver, intestine and kidney were performed to observe any abnormalities. Finally, the number of enterobacteria before and after Mo consumption was monitored.

**Results:** No significant difference was found between the amount of micronucleus on control group with groups administered with Mo. There were no adverse effects at any of the doses, so no signs of toxicity were found. A slight tendency to decrease glucose was observed with the Mo groups. As well we observed weight gain in diabetic groups that were not treated with Mo. Similar results were obtained for measurements of triglycerides, cholesterol, LDL and HDL. In intestinal microbiota no differences were observed between the Mo groups and control group. Histologically, no abnormal change was observed in the administered groups with Mo in comparison with control group.

**Conclusions:** The consumption of Mo powder at high or low doses did not represent a risk in the experimental model. Previous studies have shown that consuming of Mo helps lower blood glucose. However, in the present study the decrease was not significant. The consumption of Mo powder could function as an adjuvant in lowering cholesterol and triglycerides. It is required to compare the consumption of Mo powder and the Mo extract to evaluate the hypoglycaemic properties.

**Keywords:** Moringa oleifera, Hypoglycemic, Genotoxicity.

(1)Basile, Lívia; (1)Lima, Cláudia; (2)Oliveira, Karoline; (3)Cassuya, Candida; (4)Cruz, Analia.

(1)PhD. Nutritionist. Nutrition Course. Federal University of Grande Dourados. Brazil.; (2)Student. Nutrition Course. Federal University of Grande Dourados. Brazil.; (3)PhD. Pharmaceutical. Nutrition Course. Federal University of Grande Dourados. Brazil.; (4)Technique in nursing. Nutrition Course. Federal University of Grande Dourados. Brazil.

**Background and objectives:** Some studies have reported the beneficial effects of green banana on diseases such as diabetes, dyslipidemia and cardiovascular disease. The objective of the present study was to evaluate the influence of regular consumption of green banana flour on risk factors associated with the metabolic syndrome.

**Methods:** The study group consisted of 44 servers from a federal university, of which 23 were from the Green Banana Flour Group and 21 from the Control Group. The individuals in the Green Banana Flour Group consumed two tablespoons per day of green banana flour, and those in the Control Group consumed two tablespoons per day of cassava flour in the main meals, for eight weeks. Nutritional assessment was performed with measures of weight, height and waist circumference, three 24-hour recalls, and biochemical tests of total cholesterol and fractions, triglycerides, fasting glucose, insulin, homocysteine and C-reactive protein, before and after consumption of flours. Systemic blood pressure was also measured. Statistical analysis was performed using Student's t-test and paired t-test, with a significance level of 5%. The research was approved by the Ethics Committee in Research of the Federal University of Grande Dourados, for the protocol nº 1.074.863, 06/11/2015.

**Results:** There was no significant difference between the baseline clinical characteristics of subjects in both groups. It was observed that after the consumption of green banana flour, individuals had a significant reduction of total cholesterol (-6.8%) and LDL cholesterol (-10.4%), and increased fiber intake (33.1%). There was no significant difference in the anthropometric variables and in the systemic blood pressure in both groups, after the consumption of the flours.

**Conclusions:** It was concluded that although green banana flour did not collaborate to reduce risk factors associated with the metabolic syndrome, it presented important contributions to the health of the individuals, such as reduction of plasma levels of total cholesterol and LDL cholesterol and increased consumption of fibers, and may play an important role in disease prevention, especially dyslipidemias and cardiovascular diseases.

**Keywords:** Green banana flour. Fibers. Dyslipidemias. Cardiovascular diseases.

**Further collaborators:** This research was financed by Fundect.
# RESEARCH IN ANALYTICAL CHEMISTRY APPLIED TO NUTRITION AS A TOOL IN TEACHING

(¹)López-Donado, Lorena; (²)Saldaña, Dulce; (³)Vargas, Rodrigo.

(¹)MSc. Nutritionist. School of Nutrition. Faculty of Medicine and Health Sciences. Mariano Galvez University. Guatemala.; (²)MSc. Pharmaceutical chemist. Professor. Faculty of Medicine and Health Sciences. Mariano Galvez University. Guatemala.

**Background and objectives:** The Faculty of Medical Sciences and Health of the Mariano Gálvez University of Guatemala provides the student with high quality laboratories with state-of-the-art technology that enables productive teaching and research development. Research provides the opportunity to develop competencies for the execution of research projects that seek the resolution of health and nutrition problems of the country.

The objective was to promote creativity as well as motivate the development of applied research in the field of nutrition.

**Methods:** The research developed during the course of Analytical Chemistry applied to Nutrition, are carried out with the equipment, reagents and supplies provided by the university. The laboratories have equipment to perform the following procedures: potentiometry, gas chromatography and high performance liquid chromatography, among others. Minor laboratory equipment and a variety of glassware including Soxhlet equipment are also used to develop these processes. The research is carried out in 3 stages, generating a preliminary project, followed by a protocol, and finally, the student has at his disposal the laboratory to develop the experimental part and final report.

**Results:** Six research projects were developed to determine important analytes in the selected foods taking care of the technical quality of the processes.

The students who have taken part in the Analytical Chemistry applied to Nutrition have excelled in different areas: 3 students have presented their results by means of posters at the scientific fair of the Faculty of Medical Sciences and Health of the university; a student has been involved as part of the scientific team of a co-financing project of the National Secretariat of Science and Technology and a student has been inclined to develop his ad gradum thesis on analytical chemistry topics.

**Conclusions:** The experiences in the course of Analytical Chemistry applied to Nutrition demonstrate the importance of promoting curiosity, as well as encouraging students to formulate research proposals, from the formulation of questions related to the subjects of the regular courses, which are taught in undergraduate medical science and health care.

**Keywords:** Analytic chemistry, Clinical nutrition, Investigation, Teaching

**Conflict of Interest Disclosure:** The authors whose names are listed immediately below certify that they have NO affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers' bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest

(such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript.

MSc. Lorena López Donado; MSc. Dulce Saldaña; MSc. Rodrigo Vargas

**Further collaborators:**

Billy Knedel. I2QB3 Coordinator. Mariano Galvez University. Guatemala.

Ricardo Sanjosé. I2QB3 Director. Mariano Galvez University. Guatemala.

# ADMINISTRATION OF THREE PROBIOTIC STRAINS MODIFIES THE INTESTINAL MICROBIOTA IN AN OBESITY RAT MODEL

(¹)Plaza-Díaz, Julio; (²)Vilchez-Padal, Laura Maria; (³)Fontana, Luis;

(⁴)Robles Sanchez, Candido; (⁵)Chueca, Natalia; (⁶)Lopez Bueno, Josefa; (⁷)Garcia, Federico; (⁸)Saez-Lara, Maria Jose; (⁹)Gil, Ángel;

(³)Gomez-Llorente, Carolina.

(¹)PhD, Department of Biochemistry and Molecular Biology II, School of Pharmacy, University of Granada, Granada, Spain;

(²)MSc, Institute of Nutrition and Food Technology "José Mataix", Biomedical Research Centre. University of Granada, Granada, Spain; (³)PhD, Dept. Biochemistry & Molecular Biology II, School of Pharmacy, and Institute of Nutrition and Food Technology "José Mataix", Biomedical Research Center, University of Granada, Spain; (⁴)MSc, Dept. Biochemistry & Molecular Biology II, School of Pharmacy, and Institute of Nutrition and Food Technology "José Mataix", Biomedical Research Center, University of Granada, Spain; (⁵)PhD, Department of Microbiology, Complejo Hospitalario Universitario de Granada, Instituto de Investigación Biosanitaria (IBS), Granada 18016, España.; (⁶)BSc, Department of Microbiology, Complejo Hospitalario Universitario de Granada, Instituto de Investigación Biosanitaria (IBS), Granada 18016, España.; (⁷)PhD, Department of Biochemistry and Molecular Biology I, School of Sciences, University of Granada, Granada, España; (⁸)Ph D/Dept. Biochemistry & Molecular Biology II, School of Pharmacy, and Institute of Nutrition and Food Technology "José Mataix", Biomedical Research Center, University of Granada, Spain.

**Background and objectives:** Intestinal microbiota (IM) plays a central role in the host's health and, consequently, control of the IM is a priority to prevent a plethora of health issues. Probiotics are used in the treatment of diverse pathologies. However, evidences of their capacity to influence the IM is scarce. The aim of this work was to investigate the effects of Lactobacillus paracasei CNCM I-4034, Lactobacillus rhamnosus CNCM I-4036 and Bifidobacterium breve CNCM I-4035 on the IM of genetically obese rats.

**Methods:** Forty-eight obese Zucker-Leprfa/fa rats and 16 Zucker-lean+/fa rats were used. Eight obese and 8 lean rats were
sacrificed as baseline. The remaining 40 obese rats were randomly assigned to receive one of the 3 probiotic strains, a mixture of L. paracasei and B. breve, or a placebo by oral administration each day for 30 days. An additional group of 8 lean rats received placebo for 30 days. Fecal samples were taken to perform fluorescence in situ hybridization (FISH) and 16S rRNA gene pyrosequencing.

**Results:** FISH analysis showed significant increases in the Bacteroides genus in the rats treated with the mixture of B. breve and L. rhamnosus, and the Atopobium cluster in the B. breve group. Pyrosequencing analysis showed: 1) an increase in the Marinilabiaceae and Streptococcaceae families in the rats fed the mixture; 2) a decrease in the Lactobacillales order in the B. breve group; 3) a decrease in the Anaerostipes genus in the L. paracasei group; 4) a decrease in the Parabacteroides genus in the L. rhamnosus group. An increase in the relative abundance of the Parabacteroides and Lactobacillus genera together with a decrease in the Prevotella and Lactococcus genera were found in the obese rats fed the placebo.

**Conclusions:** These probiotic strains are capable of modifying the IM of genetically obese rats.

**Keywords:** Obesity, Probiotics, Intestinal microbiota

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PRENYLATION AT CARBON-6 RESULTS IN DECREASED TRANSEPITHELIAL TRANSPORT OF SELECTED FLAVONOIDS FROM HOPS (HUMULUS LUPULUS L.) IN CACO-2 CELLS

**Background and objectives:** 6-Prenylnaringenin (6-PN) and 8-prenylnaringenin (8-PN) are prenylflavonoids predominantly found in hops (Humulus lupulus L.) and beer, with potential health-promoting properties. Their oral bioavailability in humans is low and limits their development for clinical applications. The aim of this study was to compare the apparent permeability coefficient (Papp) of both 6-PN and 8-PN using the transepithelial

transport experiment with Caco-2 cells, a well described in vitro model of intestinal absorption, which can be correlated to in vivo absorption in humans.

**Methods:** Caco-2 cells were grown in permeable membrane inserts until enterocytic differentiation. The integrity of the epithelium was confirmed by testing the transepithelial electrical resistance and Lucifer Yellow permeability. The monolayers were incubated with 100 µmol/L 6-PN or 8-PN in the apical chamber, and the apical (lumen) to basolateral (systemic) transport of 6-PN and 8-PN was determined by HPLC. Prenylflavonoid concentrations in the apical chamber and inside the cells were also measured at the end of the experiment. Papp was calculated for both free and total (after deconjugation with β-glucuronidase/sulfatase) prenylflavonoid concentrations.

**Results:** The Papp of 8-PN was significantly higher than for 6-PN, but only when comparing the concentrations after enzymatic de-conjugation and not when comparing the free compound alone. Moreover, Papp of total 6-PN (8.4±0.7x10⁻⁶ cm/s) and total 8-PN (2.4±0.4x10⁻⁵ cm/s) were 12.7 and 38.7-fold higher than Papp of the free compound, respectively. Of the initial dose, only up to 8% total 6-PN and 25% total 8-PN were recovered in the basolateral chamber. Both compounds displayed evidence of considerable efflux of metabolites towards the apical chamber and abundant accumulation of the free substance in the enterocytes.

**Conclusions:** The prenylflavonoids 6-PN and 8-PN exhibited low intestinal permeability in vitro, with less than 1% transepithelial transport of the compounds in the free form. In addition, the significantly lower Papp for 6-PN suggests that prenylation at carbon-6 may result in decreased bioavailability of this compound in comparison to its positional isomer 8-PN. Further research should focus on the search for alternatives to improve the bioavailability of these bioactive prenylflavonoids.

**Keywords:** 6-prenylnaringenin, 8-prenylnaringenin, Humulus lupulus, Caco-2 cells, Apparent permeability.

**Further collaborators:**

This project was funded by grants from the AKF program of the Medical Faculty of the Eberhard Karls University of Tuebingen; the Wissenschaftsfoerderung der Deutschen Brauwirtschaft e.V. project B103; and the European Foundation for Alcohol Research (ERAB) Grant Ref. EA 15 28. LACC was funded by Costa Rica Institute of Technology.

<footnote>(1)Calvo-Castro, Laura A.; (2)Rojas Rodríguez, Pablo; (3)Sus, Nadine;</footnote>

<footnote>(4)Irías Mata, Andrea Paola; (5)Venturelli, Sascha; (6)Frank, Jan.</footnote>

<footnote>(1)MSc. 1. Division of Biofunctionality and Safety of Food. Institute of Biological Chemistry and Nutrition. University of Hohenheim. D-70599. Stuttgart. Germany. 2. School of Biology. Costa Rica Institute of Technology. Cartago. Costa Rica.; (2)School of Biology. Costa Rica Institute of Technology. Cartago. Costa Rica.; (3)BTA. Division of Biofunctionality and Safety of Food. Institute of Biological Chemistry and Nutrition. University of Hohenheim. D-70599. Stuttgart. Germany.; (4)MSc. Division of Biofunctionality and Safety of Food. Institute of Biological Chemistry and Nutrition. University of Hohenheim. D-70599. Stuttgart. Germany.; (5)PhD. MD. Institute of Physiology. Department of Vegetative and Clinical Physiology. Medical University Hospital. Tuebingen. Germany.; (6)PhD. Division of Biofunctionality and Safety of Food. Institute of Biological Chemistry and Nutrition. University of Hohenheim. D-70599. Stuttgart. Germany.</footnote>