EFFECTS OF NUTRIENTS AND TOXICANTS ON HUMAN HEALTH

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1017721
• Project Start Date: Oct 3, 2018
• Project End Date: Sep 30, 2023
• Program Code: [(N/A)]- (N/A)

Recipient Organization
UNIVERSITY OF CALIFORNIA, DAVIS
410 MRAK HALL
DAVIS,CA 95616-8671

Performing Department
Interdepartmental

Non Technical Summary
This project supports the mission of the Agricultural Experiment Station by addressing the Hatch Act areaof human nutrition.Undernutrition, overnutrition and exposure to toxic substances that affect nutrient availability have major adverse consequences on human health. While potentially affecting many populations, these conditions are particularly frequent in low-income groups. The food we eat and the toxic substances we are exposed to can have a major influence on our brain health and function. This project will focus on the impact of diet and environmental factors on brain development and brain inflammation and their consequences on brain function and ultimately on behavior and learning capabilities. Thus, we will study:A) The impact of low levels of the zinc (Zn) (due to a deficient diet or exposure to toxic substances) during gestation and early life on brain development and the risk of the newborns to develop neurological/behavioral problems. A large body of evidence indicates that diet and environmental exposures during pregnancy and early development have a major impact on the offspring's health and on the risk to develop diseases later in life. This is why understanding the relevance of Zn in normal brain development is of critical public health relevance to evaluate the need of preventive strategies.B) The capacity of dietary flavonoids (which are naturally occurring compounds present in fruit, vegetables, chocolate, and beverages like wine and tea) to alleviate the inflammation of the brain caused by overnutrition/obesity and how these affect behavior. We will also examine the benefits of varying the content of active dietary flavonoids by using particular fruit varieties and farming practices. Many experts have long recognized the need to identify specific fruit compounds that have biological actions in order to design better knowledgeable recommendations on not only which fruits are the healthiest, but within a fruit, which are the varieties and the cultivar/postharvesting practices that provide maximum levels of beneficial bioactives, including dietary flavonoids. This aspect of the proposal is highly relevant given the alarming impact of Western-style diets on health and its consequences on obesity-related pathologies in the USA and worldwide.

Animal Health Component

20%

Research Effort Categories

Basic

80%

Applied

20%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
702	7010	1010	60%
802	6020	1150	40%

Knowledge Area
702 - Requirements and Function of Nutrients and Other Food Components; 802 - Human Development and Family Well-Being;

Subject Of Investigation
7010 - Biological Cell Systems; 6020 - The family and its members;

Field Of Science
1010 - Nutrition and metabolism; 1150 - Toxicology;

Keywords

zinc

plasticizers

development

behavior

obesity

flavonoids

neuroinflammation

Goals / Objectives
JustificationUndernutrition, overnutrition and exposure to toxicants that affect nutrient availability have major adverse consequences on human health. This project will focus on the impact of diet and environmental factors on brain development and brain inflammation and their consequences on behavior. We will study: A) the impact of suboptimal zinc (Zn) nutrition or availability (i.e. plastizicer exposure) during gestation and early postnatal stages on brain development; and B) the capacity of flavonoids to mitigate overnutrition/obesity-induced neuroinflammation. While potentially affecting many populations, these conditions are particularly frequent in low-income groups.1. Brain development: relevance of diet and toxicant exposure affecting Zn homeostasis. Zn deficiency associated with inadequate Zn intake and undernutrition is frequent in the U.S.A. (1, 2), and worldwide (3, 4). Zn deficiency can occur not only as a dietary deficiency, but secondary to frequent conditions associated to poverty, such as chronic infection, parasitosis, toxicant exposures, alcoholism and chronic diseases (e.g. uncontrolled diabetes) (5, 6). Our long term goal is to elucidate if decreased Zn availability during early development could increase the risk to develop neurological disorders in childhood, adolescence or early adulthood. This aim will focus on the importance of adequate gestational and early postnatal Zn nutrition and on the impact of gestational toxicant exposure (i.e. di-2-ethylhexyl phthalate (DEHP)) that affect Zn homeostasis, on the optimal neurological and neurobehavioral outcome of the offspring. A better understanding of the mechanisms that contribute to the adverse effects of Zn deficiency will result in improved methods for the identification of a functional Zn deficiency, and its treatment.2. Brain inflammation: relevance of diet on the gut-brain interactions. Overnutrition, particularly the consumption of diets rich in carbohydrates and fat, constitute a major health concern worldwide given the associated risk for obesity and associated diseases. Epidemiological studies on obesity and cognition in children, adolescents, and adults provide evidence that obesity is positively associated with cognitive impairment (7-10). Based on our previous results (11-16), we will investigate the capacity of select flavonoids present in fruit and vegetables (i.e. epicatechin (EC), catechin (CT), procyanidins (PAC), anthocyanins (AC)) to mitigate the adverse consequences of consuming Western styles on brain inflammation and behavior. We will assess the benefits of varying the content of CT, PAC and AC by using particular fruit (i.e. grapes) varieties and farming practices. The field of bioactives has long recognized the need to identify specific fruit compounds that have biological actions in order to design knowledgeable recommendations on not only which fruits are the healthiest, but within a fruit, which are the varieties and the cultivar/postharvesting practices that provide maximum levels of beneficial bioactives. This aspect of proposal is highly relevant given the alarming impact of Western-style diets on health and its consequences on obesity-related pathologies in the USA and worldwide.Previous work and present outlookAim 1. Zn deficiency, toxicant exposure and brain development. The developing brain can be highly sensitive to a deficit of this nutrient for several reasons, including the need of Zn for cell differentiation, migration, and growth. Zn deprivation during critical developmental periods can result in altered emotionality and food motivation early in life (17). A moderate Zn-deficient diet imposed during the adolescent growth spurt in monkeys leads to an impairment in the performance of attention and memory tasks and in the amount of time spent active, which are reverted by Zn supplementation (18). In humans, suboptimal Zn nutrition during development is associated with altered neonatal/infant behaviour, cognitive and motor performance (19-22). In the frame of our previous 5 year HATCH, we found that both primary (dietary) and secondary (exposure to DEHP) Zn deficiency affect maternal and fetal steroid profiles (23). However, fetal exposure to glucocorticoids was not increased, and glucocorticoid metabolism by the placenta was not affected by Zn deficiency (24), suggesting that glucocorticoids are not involved in the deleterious effects of developmental Zn deficiency. Gestational Zn deficiency inhibited the STAT1/STAT3 cascades in the E19 brain (25), which irreversibly affected astrocyte number and distribution in the mature brain ((26) and paper in preparation). Gestational Zn deficiency decreased the number of brain fetal progenitors (27) which could impair neurogenesis and oligodendrogenesis affecting the number and distribution of neurons and oligodendrocytes/myelination in the adult brain. Thus, the overall objective of this aim is to investigate how developmental nutritional and secondary Zn deficiency (toxicant exposure) can disrupt neurogenesis and oligodendrogenesis leading to an increased risk for altered offspring behavior. This aim will investigate:1.1. The effects of gestational Zn deficiency on neuro and oligodendrogenesis. 1.2. The effects of early postnatal marginal Zn deficiency on precursor differentiation into mature oligodendrocytes and on myelination. 1.3. The consequences of both gestational (aim 1.1) and early postnatal (aim 1.2.) marginal Zn deficiency on the number, type and distribution of neurons and oligodendrocytes, and on myelination in the adult brain. Aim 2. Brain inflammation: relevance of diet on the gut-brain interactions. Obesity and its comorbidities are causing not only serious health burdens but also incalculable social and medical costs. While consumption of Western-style diets can lead to obesity, inflammation, steatosis, type 2 diabetes (T2D); consumption of fruits and vegetables can have protective effects (28). The gastrointestinal (GI) tract plays a central role in sustaining overall health, while disruption of its physiology can lead to disease (29). In this regard, the permeabilization of the intestinal barrier can allow the passage of bacterial endotoxins which can initiate local and systemic inflammation. Intestinal dysbiosis also contributes to endotoxemia in obesity (30), being proposed as an underlying mechanism in obesity-associated pathologies (30). Significantly, fat and energy intake are associated with endotoxemia both in healthy (31, 32) and in T2D individuals (33). Additionally, the GI tract secrete hormones that have important functions (34, 35), including the inhibition of neuroinflammation (36). Flavonoids can have multiple beneficial effects at the GI tract, stressing their potential relevance in preventing diet-induced endotoxemia and neuroinflammation. Our previous results showed that (-)-epicatechin protect rodents from high fat- and high fructose-mediated development of obesity, dyslipidemia, T2D, and adipose/liver inflammation (11, 12, 16). These beneficial effects were associated with the preservation of the gut barrier integrity and a decrease in circulating endotoxins (15). We recently showed that also AC prevent high fat-induced insulin resistance, dyslipidemia, inflammation and steatosis (37) and prevented inflammation-induced monolayer permeabilization in Caco-2 cells The overall objective of this aim is to investigate if select flavonoids (EC, PAC and AC) can prevent high fat diet-induced neuroinflammation and associated altered behavior. This aim will investigate:2.1. The capacity of EC, AC and PAC to inhibit high fat diet-induced neuroinflammation and its association to endotoxemia in mice. 2.2. The mechanisms underlying the capacity of EC, AC and PAC to modulate endotoxemia. 2.3. The capacity of EC, AC and PAC to prevent behavioral changes associated with the consumption of high fat diets and obesity in mice.

Project Methods
Experimental procedures Aim 1. 1.1.The effects of Zn deficiency during gestation on neurogenesis and oligodendrogenesis. Zn deficiency will be either induced between gestation day 0 and postnatal day 2 through dietary deficiency, or as a secondary deficiency caused by exposure to the plasticizer DEHP.1.2. The effects of early postnatal marginal Zn deficiency on OPC differentiation into mature oligodendrocytes and on myelination. A Zn deficient diet will be fed to rats between post-natal days 0 and 20, and OPC differentiation followed at postnatal days 5, 10 and 20.1.3. The consequences of both gestational (aim 1.1) and early postnatal (aim 1.2.) marginal Zn deficiency on the number, type and distribution of neurons and oligodendrocytes, and on myelination in the adult brain. Biochemical and histological parameters will be measured at postnatal day 56 and associated changes in behavior one week before.Rat model of developmental Zn deficiency. Female adult Sprague-Dawley rats will be fed an egg white protein based diet with adequate Zn (25 µg Zn/g) (control) or a marginal Zn (10 µg Zn/g) (MZD) content. Animals will be fed; A- control or MZD diets from gestation day 0 (GD0) until postnatal day 2 (P2), or B- control or MZD diets from P2-P20 (weaning). For both studies, either at P2 or P20 all animals will be fed the control diet until P56. Two additional groups (ten rats per group) will be fed the control or marginal Zn diets supplemented with 300 µg DEHP/g diet from GD0 until GD19.Characterization of neurogenesis, oligodendrogenesis and myelination. Differentiation of progenitor cells into neurons and oligodendrocytes will be followed through the expression of specific protein markers by Western blot, RT-PCR and immunohistochemistry (IHC). Myelination will be evaluated through the expression of myelin proteins by Western blot and IHC. IHC will be the methods to assess neuron, oligodendrocyte and myelin abundance and distribution.Cell culture. IMR-32 cells (human neuroblastoma) will be used to evaluate the effects of Zn deficiency on the capacity of neuronal cells to respond to oxidant stressors (dopamine) and the underlying signaling. Cells will be incubated in control or Zn-depleted media containing 1.5 15 or 50 µM Zn and exposed to different concentrations of dopamine. Cells will be incubated for variable periods of time depending on the experiment.Aim 2.2.1.The capacity of EC, AC and PAC to inhibit high fat diet-induced neuroinflammation and its association to endotoxemia in mice. Diet-induced obesity and neuroinflammation will be induced through a high fat diet fed to mice for 15 weeks. EC, AC and PAC (2-40 mg/kg body weight) will be supplemented in the diet as either single compounds, berry extracts or whole berry powders. The development of endotoxemia and protective actions of flavonoids will be also investigated after an acute fat intake.2.2.The mechanisms underlying the capacity of EC, AC and PAC to modulate endotoxemia. We will evaluate if the inhibition of endotoxemia by flavonoids occurs through the preservation of the GI barrier integrity, the modulation of LPS transport through chylomicrons, and/or by shaping the microbiota.2.3. The capacity of EC, AC and PAC to prevent behavioral changes associated with the consumption of high fat diets and obesity in mice. We will evaluate locomotor activity, exploration and anxiety, short-term and spatial memory, learning.Mouse model of high fat diet-induced obesity. Healthy male C57BL/6J mice (20-25 g) (10 mice/group) will be fed for 15 weeks either: A- a diet containing approximately 10% total calories from fat (Control) (TD.06416, Harlan Laboratories, Madison, WI), B- a diet containing approximately 60% total calories from fat (lard) (HF) (TD.06414, Harlan Laboratories, Madison, WI), C- the control and the high fat diet supplemented with EC, PAC or AC (2-40 mg/kg body weight). Body weight and food intake will be measured weekly.Acute exposure to a high fat meal. The effects of consuming a single high fat amount on endotoxemia and LPS transport via chylomicrons will be investigated inmice gavaged with triolein and LPS. The kinetics of LPS appearance in plasma and in chylomicrons will be followed. Chylomicrons synthesis and parameters of endotoxemia (LPS binding protein, TLR4) and inflammation will be evaluated.Characterization of neuroinflammation. Protein markers of neuroinflammation and of the tightly associated endoplasmic reticulum stress will be evaluated by RT-PCR and Western blot in brain cortex and hippocampus, and by IHC in the whole brain.Metabolic parameters, barrier permeability and endotoxemia will be assessed as we previously described (15).Methods common to both aimsEvaluation of signaling pathways will be done using different approaches: 1- activation of upstream events and downstream targets by Western blot, 2- nuclear transport and DNA binding activity (EMSA, Western blot), 4- expression of target genes (Western blot and RT-PCR), 3- immunohistochemistry (IHC), 4-phosphatase/kinase activity. Behavior For the assessment of the impact of Zn deficiency on behavior in the young adult rat, we will do: i) an open field test to assess animal's locomotor activity, exploration and anxiety, and ii) a rotarod test to assess sensorimotor function, balance and coordination. For the assessment of obesity-induced altered behavior we will do: i) an open field test, ii) a novel object recognition task to assess short-term object recognition memory, iii) an object location memory task to evaluate spatial memory, and iv) a Morris water maze test to assess spatial learning and reference memory.Duration We consider that the project will take approximately 5 years. We completed most of the experimental and writing aspects of the previous proposal within the 5 year time frame. All the proposed models are of current use in my laboratory.Financial SupportUnrestricted Gifts (Packer-Wentz Endowment, Pharmanex Res.)Research contract (Nuskin Ent.)Pending support: USDA NIFA-AFRI (we are resubmitting a previously (2017) rated high priority proposal)PersonnelPrincipal Investigator (30% effort): Patricia I. Oteiza, Professor, Department of Nutrition and Department of Environmental Toxicology, University of California, Davis.Postdoctoral fellows (100% effort): Eleonora CremoniniPostdoctoral fellow (100% effort): To be appointedGraduate student (50% effort): Xiuzhen LiuGraduate student (50% effort): JiYe KangGraduate student (50% effort): Ziwei WangGraduate student (50% effort): Zacchary KaufmanCollaborators:Dr. Ana M. Adamo, Associate Professor, University of Buenos Aires, Argentina.Dr. Cesar G. Fraga, Professor, University of Buenos Aires, Argentina; Affiliated Faculty, Department of Nutrition, University of California, Davis.Dr. Gerardo Mackenzie, Assistant Professor, Department of Nutrition, University of California, Davis.Dr. David Mills, Professor, Department of Viticulture and Enology, University of California, Davis.Dr. Gabriela Salvador, Assistant Professor, Universidad Nacional del Sur, Argentina.Dr. Andrew Waterhouse, Professor, Department of Viticulture and Enology, University of California, Davis.Institutional Units InvolvedDepartment of Nutrition and Department of Environmental Toxicology, College of Agriculture and Environmental Sciences, University of California, Davis.CooperationNone

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:The research in this project directly addresses problems of great importance to highly labile populations of California: pregnant women and small children, and the increasing obese population that particularly affects minorities and the poorer. We have presented the results of our research in scientific conferences, universities, and private companies that sell food and supplements. The audience of my research presentations include physicians, nurses, registered dietitians and other participants in the area of human health. They can later convey the information to their patients and communities. In my interactions with private companies I have: i) contributed to design and improve existing dietary supplements, ii) proposed research that would scientifically support the importance of their products on human health, iii) helped find ways to improve the availability of fruit/vegetable bioactives with recognized health effects, and iv) increase the commercial value of crops and their industrial byproducts. I have also presented seminars to future professionals in public health, on the impact of nutrition and toxicant exposure during early development on the risk of disease and disabilities later in life. The ultimate purpose of my research is to improve nutrition to promote health and normal development, and mitigate toxicological exposures by improving food safety and working conditions. Iam actively involved through different programs, as a Hispanic woman and faculty, on coaching minority undergraduate, graduate students and young faculty. These activities have the purpose to support and encourage them on their studies and to help them in their advancement in the next steps of their academic and/or professional careers. In this regard, I am part of the Graduate Students of Color (GSoC) Mentoring Program. As part of this program, throughout 2020 I mentored a Latino graduate student, helping her navigate the advancement of her career in the particular Covid19 circumstances, and providing socioemotional support. Although this activity lasts two quarters for each student, I continue to support her and plan to do it throughout her career at UCD. I also participated in the NUTGAP Program (Nutritional Biology Graduate Admissions Pathways) (https://nutgap.ucdavis.edu/) aimed at "increasing the number of scientists from underrepresented groups who pursue research careers in nutritional biology" In 2020 I also attended the 2020 World Ag expo CA&ES booth (February 12). As a representative of UC Davis at this event, I interacted with many families and prospective students. I informed them about the College, majors and life at UCD. I grew up in a farm, and I felt that this helped me connect even better with those families living in rural communities. I also discussed my research with those interested, including agricultural stake holders, and provided some nutritional and toxicological advice. Finally, at the end of 2020 we submitted a grant that would help us reach out to rural agricultural communities in the central valley. In partnership with the EHSC Community Engagement Core and the EHSC Community Stakeholder Advisory Committee (CSTAC) we aim to predict environmental chemicals exposures through food intake pattern analysis among migrant and seasonal farmworkers, and provide toxico-nutritional consumption advice in these populations. Changes/Problems:No problems are envisioned. What opportunities for training and professional development has the project provided?Many undergraduate and graduate students, postdoctoral fellows and international scholars usually work and advance their careers under the umbrella of this HATCH project. Given the extenuating circumstances of 2020, only the Ph. D. students and postdoctoral fellows could do experimental work. However, we managed to provide online opportunities to undergraduate students. Students and pot-docs that worked under the umbrella of this HATCH project during 2020 were: UC Davis undergraduate internships Students are usually incorporated in a research project. Given the unusual circumstances of 2020, only internships that allowed online work, i.e. evaluation of animal behavior through videotaped behavioral tests, scientific reading, oral presentations and paper writing, were incorporated as research activities. Janine Mozoomdar Vanessa Veronica Garnadi Elena Chai Cecilia Tran Yue-Liang Brewerton Melissa Clark Xianyu (Adam) Zhu Saba Almassi UC Davis NUTGAP program; Sarry Pierre. I mentored this student as part of the NUTGAP Program (Nutritional Biology Graduate Admissions Pathways) (https://nutgap.ucdavis.edu/) developed to "increasing the number of scientists from underrepresented groups who pursue research careers in nutritional biology" *Given Covid19, research and instruction was done online. He read papers and analyzed data from studies linking diet-induced obesity in mice with the development of type 2 diabetes. He gave an oral presentation showing the obtained data and his conclusions. Polytech'Montpellier-University of Montpellier/AgroSup Dijon University (France) undergraduate interns Students usually do a 5-6 months research visit, which includes a research project and a final written thesis required to obtain the professional degree of Engineers in Food Science. Given Covid19, online research and instruction was in place. Research consisted of evaluation of mouse behavior in a model of high fat diet-induced obesity. They analyzed video recordings and processed data. They wrote a paper on the topic in preparation for the final thesis. Celine Houri Laura Paper Graduate students I do not only directly advise the graduate students listed below, but I am the Master Adviser for the Graduate Group in Nutritional Biology. In 2020 I received the "Graduate Program Advising and Mentoring Award" which is given by the Office of Graduate Studies, UC Davis. Ph.D. thesis (role of P. Oteiza) Liu, XiuZhen, Ph.D. (GGNB) In progress (Chair) Wang, Ziwei, Ph.D. (GGNB) In progress (Chair) Kang, JiYe, Ph.D. (GGNB) In progress (Chair) Zhara Mostofinejad, Ph.D. (GGNB) In progress (Chair) Catherine Cox, M.S. (GGNB) Awarded (Member) Post-doctoral fellows Eleonora Cremonini Wei Zhu Dario Iglesias How have the results been disseminated to communities of interest?We have disseminated our research through: 1-the presentation of several abstracts/posters to national and international conferences. Several students/researchers were distinguished with awards and invitations for oral presentations. 2- several conferences presented by the P.I. and postdocs in academic and professional environments with direct input to health professionals. 3- publication of research papers. 4-Teaching undergraduate and graduate courses on nutrition and health not only at UC Davis but internationally (Argentina/Germany Masters program). 5-Supporting the career advancement of minority UCD students, particularly women. I have had special interactions with latino women graduate and undergraduate students given that they look at me as a role model given my Hispanic origin. Having the opportunity to help the advancement of promising young women, through advising and support for fellowships and grants, has been a very rewarding experience. 6-Giving presentations at companies interested in bioactives from agricultural origin. 7-Sharing my research with the general public and stake holders at the 2020 World Ag expo CA&ES booth. What do you plan to do during the next reporting period to accomplish the goals?I will continue the ongoing projects focused on A-the understanding of the impact of zinc undernutrition and of toxicants that could affect zinc homeostasis during gestation on brain development, focusing on myelination and motor performance; B- the interactions between zinc nutrition and toxic plasticizers on early development; C- the interactions of diet and toxicants for toxicant absorption/distribution, and D- continue to provide evidence of the beneficial health effects of phytochemicals present in US agricultural products that can provide both, a benefit for human health and also contribute evidence to add commercial value to this products and derivatives. If the submitted grant is not awarded, I will apply for other sources of financing to support our research on toxicological exposures and the impact on diet in California agricultural workers.

Impacts
What was accomplished under these goals? The focus of this project is to understand the impact nutrition and exposure to toxicants on human health at different life stages. During the current period we have worked on the following projects: A- One major aspect of my research interest is the investigation of the role of zinc in the development of the nervous system. We hypothesize that marginal zinc nutrition during critical developmental periods can affect neural progenitor cell proliferation, migration, differentiation and survival. This could affect the genesis and distribution of neurons, astrocytes and oligodendrocytes which could be one underlying cause of the altered motor and cognitive functions associated with developmental zinc deficiency. During the period under review: A.1. We published evidence that a mild maternal zinc deficiency affects the antioxidant defenses in the developing brain and in particular in neurons. We described that decreased availability of zinc make neurons more susceptible to oxidant stressors/toxicants. We advanced research on the effects of maternal marginal zinc deficiency on oligodendrogenesis and myelinogenesis in the developing offspring. Recent results show that marginal zinc deficiency during critical developmental periods in rats affects the amount of myelin in the adult offspring's nervous system, affecting motor performance. These findings are of enormous relevance, because they show that even a mild deficiency of zinc during brain development can have long-term and irreversible effects on brain structure, and potentially function. A.2. We completed studies showing that gestational exposure to plasticizers (DEHP) disrupts embryonic brain zinc homeostasis. We found similar changes in embryonic brain neurogenesis as those found in rats fed a marginal zinc diet during pregnancy. These effects and the underlying mechanisms were confirmed in neuronal cells. B- Other major aspect of my research interests is the investigation of the health effects of dietary bioactives. This research can significantly help increase the commercial value of particular fruits, vegetables and crops, and give commercial value to byproducts of the wine industry (grape pomace). We did research on flavan-3-ols, procyanidins, anthocyanins and ellagic acid that are present, among others, in grapes, apples, berries, persimmons, pomegranates, nuts and select rice varieties. In this regard: B.1. We continued investigating the mechanisms of action of the flavonoid (-)-epicatechin (EC) as anti-inflammatory/anti diabetic bioactive. With all the students and post docs in my laboratory we published a review on the beneficial actions of dietary epicatechin against the comorbidities of obesity. We also published evidence that EC supplementation can attenuate high fat diet-induced neuroinflammation in mice and improve associated alterations in behavior. B.2. We published a relevant study showing the potential anticolorectal cancer actions of persimmon dimeric procyanidins. These studies provide strong support to the concept of a beneficial capacity of fruit and vegetable consumption in decreasing colorectal cancer risk. B.3. We published evidence that grape pomace and some of its components help attenuate adipose hypertrophy occurring as a consequence of consumption of high fat diets in rats. The grape pomace also promoted the conversion of white adipose tissue into a healthier and "browner" tissue through the induction of mitochondriogenesis. Such effects have a direct beneficial impact against the negative consequences of an enlarged and inflamed adipose tissue on human health. B.4. We demonstrated the capacity of ellagic acid to mitigate inflammation in a cell model of intestinal epithelium and prevent its permeabilization. This is very relevant given that intestinal permeabilization is a major contributor to obesity-induced chronic inflammation and the associated comorbidities, e.g. diabetes, cardiovascular disease and non-alcoholic fatty liver disease. Results described in section A can help raise awareness on the relevance of maternal/child nutrition on brain development, cognition and behavior. Results described in section B not only support important health benefits of select bioactives, but can also be of high relevance from an agriculture stand point, providing growers with information on particular crops, varieties and cultivation conditions that help them develop new options/markets for their products.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: Kaufman, Z., Salvador, G.A., Liu, X.,�Oteiza, P.I. Zinc and the modulation of Nrf2 in human neuroblastoma cells. Free Radical Biology and Medicine (2020) 155:1-9.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Cremonini E, Iglesias, D. E., Kang, J., Lombardo, G.E., Mostofinejad, Z., Wang, Z., Zhu, W. and �Oteiza, P.I. (-)-Epicatechin and the comorbidities of obesity. Archives of Biochemistry and Biophysics (2020) 690:108505
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Perdicaro, D.J.. Rodriguez Lanzi, C., Gambarte Tudella, J., Miatello, R.M., Oteiza, P.I., Vazquez Prieto, M.A. Quercetin attenuates adipose hypertrophy, in part through activation of adipogenesis in rats fed a high-fat diet Journal of Nutritional Biochemistry (2020) 79:108352
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Iglesias, D. E., Cremonini, E., Fraga, C.G., Oteiza, P.I. Ellagic acid protects Caco-2 cell monolayers against inflammation_induced permeabilization. Free Radical Biology and Medicine (2020) 152:776-786
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Rodriguez Lanzi, C., Perdicaro, D.J.. Gambarte Tudella, J., Muscia, V. Fontana, A.R., Oteiza, P.I., Vazquez Prieto, M.A. Grape pomace extract supplementation activates FNDC5/irisin in muscle and promotes white adipose browning in rats fed a high-fat diet. Food and Function (2020) 11:1537-1546
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Zhu, W., Li, C., Wang, R., Mackenzie, G.G., and Oteiza, P.I. The inhibitory effect of ECG and EGCG dimeric procyanidins on colorectal cancer cells growth is associated with their actions at lipid rafts and the inhibition of the Epidermal Growth Factor Receptor signaling. Biochemical Pharmacology (2020) 175:113923
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Kang, J., Wang, Z. and Oteiza, P.I. (-)-Epicatechin mitigates high fat diet-induced neuroinflammation and altered behavior in mice. Food and Function (2020) 11:5065-507

Progress 10/03/18 to 09/30/19

Outputs
Target Audience:The research in this project directly addresses problems of great importance to highly labile populations of California: pregnant women and small children, and the increasing obese population. We have presented the results of our research in scientific conferences, universities, and private companies that sell food and supplements. The audience of my research presentations include physicians, nurses, registered dietitians and other participants in the area of human health. They can later convey the information to their patients and communities. In my interactions with private companies, I have contributed to design and improve existing dietary supplements. I have also presented seminars to future professionals in public health, on the impact of nutrition and toxicant exposure during early development on the risk of disease and disabilities later in life. I am actively involved through different programs, as a Hispanic woman and faculty, on coaching high school, undergraduate and graduate students outside the classroom. This has the purpose to support and encourage them on their studies and on their advancement in the next steps in their academic or professional careers. Changes/Problems:No problems are envisioned. What opportunities for training and professional development has the project provided?Many undergraduate and graduate students, postdoctoral fellows and international scholars have worked and advanced in their careers under the umbrella of this HATCH project, including: UC Davis undergraduate internship Students are incorporated in a research project and in some cases, they presented posters and/or their work was part of a publication. Promise Lee (Nutritional Sciences Major) (2018-2019) (presented poster at the UC Davis URC Undergraduate Conference) Zachary Booze (Global Disease Biology) (2019) I supervised his practicum on "Meta-analysis of Micronutrient Malnutrition, Undernutrition, and The Factors That Contribute to Them" , a requirement to complete his graduation. Research Internships (2 quarters): Janine Mozoomdar Vanessa Veronica Garnadi Elena Chai Cecilia Tran Yue-Liang Brewerton Melissa Clark Xianyu (Adam) Zhu Saba Almassi Polytech'Montpellier-University of Montpellier/AgroSup Dijon University (France) undergraduate interns Students do a 5-6 months research visit, which includes a research project and a final written thesis required to obtain the professional degree of Engineers in Food Science. Pierrick Girard (2019) Pauline Pinel (2019) Ph.D./M.S. thesis (role of P. Oteiza) Kauffman,Z. M.S. (GGNB) Awarded (Chair) Liu, XiuZhen, Ph.D. (GGNB) In progress (Chair) Wang, Ziwei, Ph.D. (GGNB) In progress (Chair) Kang, JiYe Ph.D. (GGNB) In progress (Chair) Chodur, G. Ph.D. (GGNB) In progress (Member) Post-doctoral fellows Eleonora Cremonini Giovani Lombardo Wei Zhu Dario Iglesias Visiting scholars Ana M Adamo, Associate Professor, University of Buenos Aires, Argentina Corina Litterio, Research Scientist, University of Buenos Aires, Argentina Mirko Marino, Graduate StudentUniversity of Milan, Italy How have the results been disseminated to communities of interest?We have disseminated our research through: 1-the presentation of several abstracts/posters to national and international conferences. Several students/researchers were distinguished with awards and invitations for oral presentations. 2- several conferences presented by the P.I. and postdocs in academic and professional environments with direct input to health professionals. 3- publication of research papers. 4-Teaching undergraduate and graduate courses on nutrition and health not only at UC Davis but internationally (Argentina/Germany Masters program). 5-Supporting the career advancement of minority UCD students, particularly women. I have had special interactions with latino women graduate and undergraduate students given that they look at me as a role model given my Hispanic origin. Having the opportunity to help the advancement of promising young women, through advising and support for fellowships and grants, has been a very rewarding experience. 6-Giving presentations at companies interested in bioactives from agricultural origin. What do you plan to do during the next reporting period to accomplish the goals?I will continue the ongoing projects focused on A-the understanding of the impact of zinc undernutrition during gestation on brain development, ifocusing on myelination and motor perfromance; B- the interactions between zinc nutrition and toxic plasticizers on early development; C- the interactions of diet and toxicants for toxicant absorption/distribution, and D- continue to provide evidence of the beneficial health effects of phytochemicals present in US agricultural products that can provide both, a benefit to human health and also contribute evidence to add commercial value to this products and derivatives.

Impacts
What was accomplished under these goals? The focus of this project is to understand the impact nutrition and exposure to toxicants on human: i) early development and ii) onset/progression of metabolic diseases. During the current period we have worked on the following projects: A- One major aspect of my research interest is the investigation of the role of zinc in the development of the nervous system. We hypothesize that a decreased zinc availability, secondary to nutrition deficits or toxicant exposures, during critical developmental periods can affect neural progenitor cell proliferation, migration, differentiation and survival. This could affect the genesis and distribution of neurons, astrocytes and oligodendrocytes which could be one underlying cause of the altered motor and cognitive functions associated with developmental zinc deficiency. During the period under review: A.1. We showed that a mild maternal zinc deficiency causes a lower number of stem cells in the fetal rat brain. Even when dams and offspring were fed a diet with normal levels of zinc postnatally, in the adult offspring cortex we observed a lower number of neurons and alterations in the specification of neurons (no effects on gabaergic neurons, and a decreased amount of cortical glutamatergic neurons). These findings are of enormous relevance, because they show that even a mild deficiency of zinc during brain development can have long-term and irreversible effects on brain structure, and potentially function. A.2. We observed (unpublished results) that gestational exposure to plasticizers (DEHP) disrupts embryonic brain zinc homeostasis. We found similar changes in embryonic brain neurogenesis as those found in rats fed a marginal zinc diet during pregnancy (See A.1.). A manuscript is in preparation to be submitted to Toxicological Sciences. B- Other major aspect of my research interests is the investigation of the health effects of dietary bioactives. During the period under review we have particularly focused our interest on the beneficial actions at the gastrointestinal tract of flavonoids (flavan-3-ols and anthocyanins), procyanidins and other plant bioactives (glucosinolates) present in grapes and other berries, select rice varieties, persimmons and kale. In this regard: B.1. We continued investigating the mechanisms of action of the flavonoid (-)-epicatechin (EC) as anti-inflammatory/anti diabetic bioactive. While EC did not affect the increased fecal amount and altered profile of bile acids in mice fed a high fat diet, it did prevent intestinal permeabilization. We found that EC prevent the permebilization of intestinal cell monolayers induced by bile acids. This could in part explain the intestinal barrier protective actions of EC observed in mice. Prevention of intestinal permeabilization can contribute to the EC's capacity to mitigate the associated endotoxemia, systemic inflammation and development of insulin resistance. B.2. We completed the study on the anti-colorectal cancer actions of persimmon dimeric procyanidins. A paper was recently submitted to Biochemical Pharmacology. B.3. We published evidence that anthocyanins can protect mice from high fat diet-induced intestinal permeabilization and associated endotoxemia. While the high fat diet caused major changes in microbiota profiles, supplementation with anthocyanins recovered the control healthy microbiota. These actions can in part explain their beneficial effects on obesity-associated insulin resistance. B.4. We assessed the bioavailability of glucosinolates from Kale in a collaboration with the Food Science Department. Glucosinolates have very important health benefits and understanding their bioavailability from kale, can make this vegetable more appealing to consumers, and benefit producers. Results described in section A can help raise awareness on the relevance of maternal/child nutrition and toxicant exposures on brain development, cognition and behavior. Results described in section B not only support important health benefits of select bioactives, but can also be of high relevance from an agriculture stand point, providing growers with information on particular crops, varieties and cultivation conditions that help them develop new options/markets for their products.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Cremonini, E., Fraga, C.G., Oteiza, P.I. (-)-Epicatechin in the control of glucose homeostasis: Involvement of redox-regulated mechanisms. Free Radical Biology and Medicine (2019) 130:478-488
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Adamo, A.M., Liu X.,Mathieu, P., Nuttall, J.R., Supasai S., and Oteiza, P.I. Early Developmental Marginal Zinc Deficiency Affects Neurogenesis Decreasing Neuronal Number and Altering Neuronal Specification in the Adult Rat Brain. Frontiers in Cellular Neuroscience (2019) 3:62
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Cremonini, E., Daveri, E., Mastaloudis, A, Adamo, A.M., Hester, S.N., Wood, S.M., Mills, D., Kalanetra, K., Fraga, C.G., and Oteiza, P.I. Anthocyanins protect the gastrointestinal tract from high fat diet-induced alterations in redox signaling, barrier integrity and dysbiosis. Redox Biology (2019) 26:101269
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Hwang, E.S., Bornhorst, G.,�Oteiza, P.I., Mitchell, A.E. Assessing the fate and bioavailability of glucosinolates in kale (Brassica oleracea) using simulated human digestion and Caco-2 cell uptake models. Journal of Agriculture and Food Chemistry (2019) 67:9492-9500
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Wang, Z., Litterio, C., Miller, M., Vauzour, D., and Oteiza, P.I. (-)-Epicatechin and NADPH oxidase inhibitors prevent bile acid-induced Caco-2 monolayer permeabilization through ERK1/2 modulation. Redox Biology (2020) 28:101360