Objective To measure biomarkers of lipid metabolism in response to a marginal zinc depletion, repletion, and supplementation in healthy male subjects. Methods Eighteen male subjects between ages of 18–45 consumed a controlled diet (80% carbohydrate, 10% fat, 10% protein) with two levels of dietary zinc. Phase I: Low zinc: 6 mg/day with 1.5 mg of phytate (2 weeks); Phase II: Zinc repletion: 10 mg/day with no phytate (4 weeks). Thereafter, a 25 mg/day zinc supplement was administered with an ad libitum diet for 3 weeks (Phase III). Targeted analysis of lipids and lipoprotein particle size were performed using standard methodology. Results Plasma zinc levels remained unchanged during Phases I and II in all subjects; they increased by 17.4 ± 3.8% (mean ± SE) at the end of supplementation (phase III). Apo‐A1 decreased significantly from 119.2 ± 13.2 mg/dL (mean ± SD) to 110.2 ± 13.2 mg/dL (p=0.003) at the end of the low zinc period. Apo‐A1 remained low at the end of phase II, but returned back to baseline after supplementation. A similar pattern was observed in HDL‐c and HDL‐L (large buoyant HDL particles) levels across the three phases. Plasma triglyceride levels increased during the 6‐week high‐carbohydrate diet, but were not modified by shifts in dietary zinc. Based on our previous observation of higher DNA strand breaks in the low zinc metabolic period and literature reports linking cancer to low HDL‐c levels, we performed a correlation analysis of DNA strand breaks and HDL subspecies. Our results show that the percent changes in HDL‐L and DNA strand breaks from phase I to II are negatively correlated (r=−0.45; p=0.05). Conclusion Results from our studies show that low zinc combined with a high carbohydrate diet increased biomarkers of dyslipidemia, but provision of adequate dietary zinc or a zinc supplement for a short period of time mitigated the increase in these biomarkers. Support or Funding Information Harvest Plus

BACKGROUND:Food fortification has been recommended to improve a population's micronutrient status. Biofortification techniques modestly elevate the zinc content of cereals, but few studies have reported a positive impact on functional indicators of zinc status.OBJECTIVE:We determined the impact of a modest increase in dietary zinc that was similar to that provided by biofortification programs on whole-body and cellular indicators of zinc status.DESIGN:Eighteen men participated in a 6-wk controlled consumption study of a low-zinc, rice-based diet. The diet contained 6 mg Zn/d for 2 wk and was followed by 10 mg Zn/d for 4 wk. To reduce zinc absorption, phytate was added to the diet during the initial period. Indicators of zinc homeostasis, including total absorbed zinc (TAZ), the exchangeable zinc pool (EZP), plasma and cellular zinc concentrations, zinc transporter gene expression, and other metabolic indicators (i.e., DNA damage, inflammation, and oxidative stress), were measured before and after each dietary-zinc period.RESULTS:TAZ increased with increased dietary zinc, but plasma zinc concentrations and EZP size were unchanged. Erythrocyte and leukocyte zinc concentrations and zinc transporter expressions were not altered. However, leukocyte DNA strand breaks decreased with increased dietary zinc, and the level of proteins involved in DNA repair and antioxidant and immune functions were restored after the dietary-zinc increase.CONCLUSIONS:A moderate 4-mg/d increase in dietary zinc, similar to that which would be expected from zinc-biofortified crops, improves zinc absorption but does not alter plasma zinc. The repair of DNA strand breaks improves, as do serum protein concentrations that are associated with the DNA repair process. This trial was registered at clinicaltrials.gov as NCT02861352.

ObjectiveTo determine the effects of marginal zinc deficiency and repletion on essential fatty acid, sphingolipid, and lipoprotein metabolism.MethodsSixteen apparently healthy male subjects between ages of 18–45 were subjected to three sequential phases of dietary zinc intake modulation: Phase 1: Low Zinc intake: 6 mg/day with 1.5 g of phytate (2 wks); Phase II: Zinc Repletion: 10 mg/day with no phytate (4 wks); Phase III: Zinc Supplementation: Ad lib diet supplemented with 20 mg zinc (2 wks). Subject weight, and compliance with diet were monitored every 3–4 days during the first two phases. Plasma free fatty acids and sphingolipids were measured by mass spectrometry. FADS1 and FADS2 activities were calculated by g‐linolenic (GLA)/linolenic acid (LA) or arachidonic acid (AA)/dihomo‐g‐linolenic acid (DGLA) ratios, respectively.ResultsPlasma zinc levels remained unchanged during Phases I and II in all subjects and increased only at the conclusion of phase III. FADS1 activity significantly (p=0.005) decreased from 5.2 ± 0.5 to 3.6 ± 0.5 (mean ± SEM) following Zn depletion. In contrast, FADS2 activity was insensitive to effects of low zinc intake. As a consequence of FADS1 activity loss, concentrations of AA‐containing phosphatidylcholine (PC), phosphatidylethanolamine (PE), sphingomyelins and plasmalogen antioxidant lipids decreased significantly. Furthermore, plasma concentrations of C16:0 ceramide increased following Zn depletion (p = 0.06) and was inversely correlated with FADS1 activity (p = 0.04; r2 = 0.6). Zn depletion also increased plasma triglyceride (TG) by 26 ± 15% (p< 0.05) and decreased high‐density lipoprotein (HDL) by 18.5 ± 3%; P<0.05. Interestingly, changes in FADS1 activity observed during the depletion was not normalized by dietary Zn repletion (Phase II) but, it was completely normalized with Zn supplementation (20 mg/day; Phase III).ConclusionDietary Zn depletion (6 mg/day) decreases FADS1 activity and lowered AA incorporation into major lipid sub‐classes. Loss in FADS1 activity was inversely correlated with C16:0 ceramide and was further associated with dyslipidemia as evidenced by significant alterations in plasma TG and HDL. These effects were not corrected with dietary Zn repletion (10 mg/day); supplementation with 20 mg Zn/d for an additional 3 weeks was required. These data suggest that FADS1 activity may be a sensitive biomarker of inadequate zinc intake and they also implicate marginal Zn intake as a novel risk factor for dyslipidemia and insulin resistance.Support or Funding InformationHarvest Plus, NIH S10OD0018070‐01

Background: The DASH (Dietary Approaches to Stop Hypertension) dietary pattern, which is high in fruit, vegetables, and low-fat dairy foods, significantly lowers blood pressure as well as low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol. Objective: The study was designed to test the effects of substituting full-fat for low-fat dairy foods in the DASH diet, with a corresponding increase in fat and a reduction in sugar intake, on blood pressure and plasma lipids and lipoproteins. Design: This was a 3-period randomized crossover trial in free-living healthy individuals who consumed in random order a control diet, a standard DASH diet, and a higher-fat, lower-carbohydrate modification of the DASH diet (HF-DASH diet) for 3 wk each, separated by 2-wk washout periods. Laboratory measurements, which included lipoprotein particle concentrations determined by ion mobility, were made at the end of each experimental diet. Results: Thirty-six participants completed all 3 dietary periods. Blood pressure was reduced similarly with the DASH and HF-DASH diets compared with the control diet. The HF-DASH diet significantly reduced triglycerides and large and medium very-low-density lipoprotein (VLDL) particle concentrations and increased LDL peak particle diameter compared with the DASH diet. The DASH diet, but not the HF-DASH diet, significantly reduced LDL cholesterol, HDL cholesterol, apolipoprotein A-I, intermediate-density lipoprotein and large LDL particles, and LDL peak diameter compared with the control diet. Conclusions: The HF-DASH diet lowered blood pressure to the same extent as the DASH diet but also reduced plasma triglyceride and VLDL concentrations without significantly increasing LDL cholesterol. This trial was registered at clinicaltrials.gov as NCT01404897.

The project reported here served to assess a curriculum for EFNEP to ensure theory compliance and content validity. Adherence to Adult Learning Theory and Social Cognitive Theory tenets was determined. A curriculum assessment tool was developed and used by five reviewers to assess initial and revised versions of the curriculum. T-tests for differences in mean responses from initial review to follow-up for each tenet and Cronbach's α for internal consistency of each tenet were conducted. Reviews found that the Eating Smart • Being Active curriculum successfully incorporated tenets of both theories and content remained true to Dietary Guidelines.

Zinc deficiency continues to be a major concern of international nutrition, yet there are still few sensitive and reliable biomarkers for identifying moderate zinc deficiency or monitoring changes in zinc status. Plasma zinc content is used most frequently, but it is influenced by conditions other than zinc intake and, therefore, is not reliable except when zinc intake is very low. In this study, the sensitivity of leukocytic zinc transporter gene expression and genomic integrity were evaluated in a moderate depletion/repletion study. Eighteen healthy men were provided a low zinc (6 mg/day) diet with added phytate for two weeks followed by an adequate zinc (10 mg/day) diet for four weeks. They then consumed an ad libitum diet while taking a 25mg zinc supplement for three weeks. Leukocyte DNA damage increased (p<0.0001) during the depletion phase, as detected by the Comet Assay. ZIP1, ZIP4, and ZnT1 zinc transporter gene expression was evaluated in isolated leukocytes and measured using qPCR. ZnT1 gene expression increased (p=0.02) during the depletion phase, although total plasma and leukocyte zinc levels did not change. These results suggest that a short‐term, low zinc intake alters genomic integrity and zinc transporter expression, but not plasma zinc. These potential biomarkers may be useful to evaluate zinc deficiency and the effectiveness of zinc interventions in conditions where plasma zinc remains unchanged.Grant Funding Source: This study was funded by a grant from HarvestPlus.

Animal and human studies show that intracellular zinc levels are sensitive to changes in dietary zinc. The protein metallothionein (MT) is maintained in proportion to intracellular zinc content, and therefore may be a useful marker of zinc status. Measuring MT levels is difficult due to the unique properties of the protein, so most studies focus on MT gene expression. Our objective was to develop new methods to quantify MT protein and MT‐bound zinc content. Zinc deficiency or excess was modeled in the Jurkat leukocytic cell line using the zinc‐selective chelator TPEN or exogenous zinc sulfate, respectively. Cellular protein was separated using size‐exclusion filtration to isolate an MT‐rich fraction. Then, endogenous zinc content was released, apo‐MT was loaded with cadmium, and MT‐bound cadmium levels were quantified by ICP; this method permitted the determination of cellular MT‐bound zinc content (MBZC) and MT zinc saturation (%MTsat). Gene expression of the major MT and zinc transporter isoforms were measured in parallel by RT‐PCR, which reflected the change in zinc balance in these cells. MBZC and %MTsat levels correlated with intracellular zinc content and changes in MT gene expression, showing bimodal responses to zinc deficiency and zinc excess. These measures of the cellular zinc‐metallothionein ratio may have utility as markers of zinc homeostasis. This work was supported by the HarvestPlus.

To determine (1) whether EFNEP participants are using thermometers, and (2) whether giving EFNEP participants a thermometer affects thermometer use.

The purpose of including physical activity (PA) in a new EFNEP curriculum, Eating Smart; Being Active (ESBA), was to teach participants the importance of regular physical activity as part of a healthy lifestyle and to teach participants simple, inexpensive, safe exercises to incorporate into their daily lives.

The Expanded Food and Nutrition Education Program (EFNEP) is a unique program funded by the USDA designed to assist limited‐resource audiences in acquiring the knowledge, skills, attitudes, and behavior necessary for improved health. The program is intended to contribute to their personal development and the improvement of the total family diet and nutritional well‐being. California is host to a diverse population and a collaborative effort over several years has resulted in an adaptation of a series of lessons that has overcome linguistic and cultural challenges. The objective of this research project was to deliver a culturally relevant adaptation of the EFNEP curriculum and examine outcomes, using a revised evaluation instrument. The lessons included a learner‐centered approach that was complemented with video clips on meal planning, increasing physical activity, reducing sugar intake and promoting families eating together. The lessons were delivered by bilingual, Hmong paraprofessionals. Seventy‐five participants completed the program. Results included gaining practical skills in food handling and safety and improvement in self‐reported behavior changes, including “planning ahead” for meals and saving money when food shopping. This project serves as a successful model with potential for reaching other underserved diverse populations.

Current recommendations, nationally and internationally, are to eat more vegetables and fruits. Improving the intake of vegetables and fruits remains an important public health challenge, especially for children. The objective of this observational, cross sectional study was to determine vegetable intake in a sample of multi‐ethnic children and examine the influence of flavor enhancers on vegetable intake. The setting was 2 elementary schools in northern California in the spring of 2006. The sample included 181 third and fourth graders of mixed ethnicities. Children were served a 2‐ounce portion of 5 different vegetables, every other day for 2 weeks, with a one‐week break between the two‐week periods for a total of 15 exposures in approximately 10 weeks. Vegetables were served plain and then with two different flavor enhancers. There were no significance differences in vegetable intake based on school, grade, ethnicity and gender. A period effect was found in total amount change. As a result, we used data from the first two periods and modeled the amount of vegetables eaten to determine any differences in the three groups while controlling for gender, school and ethnicity. There was a significant difference between vegetable intake alone versus vegetable with flavor enhancers(p<.0001). For romaine, broccoli and celery, there was a significant difference with flavor enhancer one (p<.003), and for green beans with flavor enhancer two (p=.0003). This study has implications for improving vegetable intake in children.

In response to Hmong community concerns regarding rising rates of chronic diseases, such as diabetes, an extender-delivered health promotion project was undertaken. From the beginning, efforts have been made to enlist the community's participation. This is an integral component of our successful outreach in each step of the project's development, delivery, and evaluation. A basic premise has been that expertise resides within the community on their own social and cultural environment, which has been shown to have a significant impact on community health. Focus groups were conducted with Hmong community members to elicit information about effective health messages. Education materials were developed, field-tested and integrated with Hmong cultural knowledge, values, and strengths. The materials and methods were integrated as part of a Hmong-adapted nutrition education curriculum. Respect was paid to a preference for learning through “showing” rather than “telling”. Taking the community seriously and engaging their participation in this way will ensure that subsequent outreach efforts are recognized as a true benefit and as a shared commitment. An extender-based project has the potential to nurture and strengthen community-centered actions that are then capable of creating significant impact in the everyday lives of the Hmong. This research was funded in part by NIH Center for Minority Health Disparities Grant Center # P60MD00222.

Links between poverty and obesity may be mediated by food purchase decisions. Price and the family not feeling hungry are two leading motivators of food purchase decisions. The Dietary Guidelines (DG) provide recommended servings but do not consider purchase price. The Food Stamp (FS) Program uses the Thrifty Food Plan (TFP) to support access to nutritious diets on a limited budget. The TFP is adjusted for changes in national budget but not regional differences and few families receive the maximum allocation. Although the DG and TFP provide unit amounts for consumption, FS allocations do not consider original supermarket unit amount that must be initially purchased. This study examined actual market food unit prices required to meet DG and TFP for a family of four for one week. Prices were collected in four California counties in a chain supermarket and a market frequented by FS recipients. Food unit prices varied slightly by county and type of market with most variation being foods with short shelf-lives. The average monthly FS allotment in California for a family of four is $178 and it is evident in all counties, the price of actual market units to comply with DG and the TFP would meet or exceed average monthly allotments within 2 weeks. This may impact food purchase decisions to less nutritious sources during the last weeks of the food stamp cycle.

The purpose of this research project was to pilot test a culturally appropriate Hmong nutrition education curriculum, adapted to the existing California Expanded Food and Nutrition Education Program (EFNEP). The program is unique in that Hmong community members provided input in the curriculum development and were recruited from three counties to implement the 8-week program. The extenders are involved in the recruitment, delivery of the lessons and the evaluation. The curriculum emphasizes healthy eating, messages from the new Dietary Guidelines for Americans 2005 and interactive learning activities. Each lesson was designed to incorporate cultural foods and recipes, Hmong-adapted handouts and culturally tailored messages. Topics covered include maintaining healthy traditional habits, MyPyramid messages, modifying less healthy food patterns, keeping foods safe and increasing physical activity. Essential components of the lessons are family involvement, goal setting and weekly evaluation. Evaluation for the program includes formative evaluation as well as use of the standard EFNEP evaluation tools, including the food behavior checklist and 24-hour recalls. Community health workers, or more specifically, extenders, who represent and work with communities provide links between providers and the broader community and have the ability to bridge cultural gaps and assist community members in making changes.

INTRODUCTION:Research indicates that low fruit and vegetable intake is a risk factor for many chronic diseases. Despite large-scale education campaigns, the great majority of Americans do not consume recommended levels. We tested the ability of a single brief interactive experience of the Little by Little CD-ROM to increase fruit and vegetable intake in low-income women.METHODS:A randomized placebo-controlled, parallel-group trial included 481 low-income, female participants: mean age 50.1 years, 48.4% African American, 51.6% non-Hispanic white, and 92.5% below 185% of the federally designated poverty level. Participants received one of three conditions: 1) a one-time experience with the Little by Little CD-ROM, 2) the Little by Little CD-ROM plus two reminder telephone calls, or 3) a stress management CD-ROM (control condition). We assessed baseline and follow-up dietary intake with a modified 24-hour recall.RESULTS:Two months after the one-time experience with the CD-ROMs, both intervention groups reported significantly higher intakes of fruits and vegetables than the control group. The Little by Little group with reminder calls increased daily intake by 1.32 fruits/vegetables, an 86% greater increase than the control group (P = .016). The Little by Little group without reminder calls increased daily intake by 1.20 fruits/vegetables, a 69% greater increase than the control group (P = .052). Significantly greater movement in Stage of Readiness for Change also occurred in the Little by Little groups compared with the control group.CONCLUSION:The Little by Little CD-ROM may be useful in public health and clinical situations to increase fruit and vegetable intake.

Previous studies suggest that rapidly exchanging zinc pools (EZP), thought to supply the zinc required by tissues, are smaller and turn over more rapidly in individuals with lower zinc intakes. We studied the effects of low dietary zinc (4.6 mg/d) on EZP mass and turnover in seven healthy men confined during a 20-wk clinical study. Supplements of 9.1 mg zinc were given during the 5-wk baseline and repletion periods, and placebos were given during a 10-wk zinc-restriction period. Stable Zn tracers were administered intravenously at the end of baseline, 3 and 10 wk after the start of zinc restriction and at the end of repletion. Multiple plasma samples were collected over an 8-d period after tracer administration. Zn:Zn ratios were measured using inductively coupled plasma mass spectrometry, and tracer-tracee data were analyzed by compartmental modeling. Activities of the zinc-dependent enzymes, alkaline phosphatase and 59nucleotidase, were unchanged during the study. There were no significant changes in EZP masses or kinetic parameters. A three-compartment model indicated that the masses of plasma zinc and total EZP averaged 3.25 6 0.58 and 147.8 6 33.2 mg, respectively, at the four time points studied. Plasma zinc mass turned over at an average of 5.3 times per hour. There was an 11% reduction (P 5 0.06) in plasma zinc flux 3 wk after the start of the low zinc diet period, but it returned to baseline values after 10 wk of zinc restriction. The results suggest that total EZP mass is maintained when dietary zinc is reduced to 4.6 mg/d over a 10-wk period. J. Nutr. 131: 2288–2294, 2001.

A dietary requirement is defined as the lowest continuing intake of a nutrient that for a specified indicator of adequacy, will maintain a defined level of nutriture in an individual. An essential dietary component is one that the body cannot synthesize in sufficient quantities to maintain health. Recommended dietary allowances (RDAs) are based on estimates of the dietary requirements, and are designed to prevent deficiency diseases and promote health through an adequate diet. In 1996, the Food and Nutrition Board (FNB) began a revision process of the RDAs using as criteria specific indicators of adequacy and functional end points for reducing the risk of chronic disease. Boron (B) is a dietary component, and evidence from animal studies indicates that it is a dietary essential; it cannot be synthesized in tissues, and organisms exposed to very low levels of B show developmental defects. In humans, there is evidence of homeostatic regulation of B and an interrelationship with bone metabolism. To understand better the relationship between dietary B and B homeostasis, we measured the dietary B intake and urinary B losses in seven male participants of a controlled metabolic study of Zn homeostasis. Average dietary B intake for the repeated menu days, days 1, 2, and 3, was 4.56, 1.87, and 4.75 mg/d, respectively. Urinary B excretion during the 42-d collection period averaged 3.20 ± 0.41 mg/d. When dietary B was low, urinary B loss (2.92 mg/d) was significantly lower than when B intake was higher (3.15 and 3.54 mg/d). Our study showed that urinary B excretion changes rapidly with changes in B intake, indicating that the kidney is the site of homeostatic regulation. To enable establishment of a dietary requirement for B in the future, further research of homeostatic regulation and functional markers of B metabolism need to be performed, followed by epidemiological studies to identify health conditions associated with inadequate dietary B.