Brief Summary of Minutes of Annual Meeting

Brief Summary of Minutes of Annual Meeting: Members Attending: Tammy Bray (Administrative Advisor), Terry Shultz (WA), Sung Koo (CT), Connie Weaver (IN), Etta Saltos (CSREES), Barbara Stoecker (OK), Raymond Glahn (NY, invited participant), Emily Ho (OR), Denis Medeiros (KS), Joy Winzerling (AZ), Cynthia Thomson (AZ, interested in joining), Mary Camire (ME), Judy Driskell (NE), Barry Shane (CA-B). Barry Shane, Chair, called the meeting to order. All attendees introduced themselves. Barry asked for a volunteer to take minutes and Terry Shultz volunteered. Etta Saltos, CSREES, presented an overview and update of the CSREES and Nutrition Programs. Etta started her presentation by outlining the NRI Competitive Grants Program which includes: * Bioactive Food Components for Optimal Health * Human Nutrition and Obesity * Improving Food Quality and Value Other nutrition related multistate research and pending nutrition-related research projects were mentioned. She reminded participants that CSREES will require electronic submission for all proposals submitted to the NRI and SBIR programs in 2007. Tammy Bray gave the Administrative Report. Tammy stressed the importance of establishing greater collaboration within the technical committee, particularly in relationship to competitive multistate grants. Tammy indicated that the annual report was due within 60 days after our meeting. The group talked about the website that had been developed. The group understood that Ann Bock had set-up a website for this project. However, the New Mexico website was not current and an updated website was needed. Members then unanimously approved a motion that the website needed updating. Because of the difficulty for any individual member to maintain a website the group then approved a motion to shift responsibility for the website from New Mexico to the USDA maintained website. Tammy Bray indicated that her office would help set up an updated website and that a staff member at Oregon State University would assist as the web master to keep the link updated with appropriate materials. The W1002 group should e-mail materials to Tammy at Tammy.Bray@oregonstate.edu. We thank Ann Bock for her contribution in maintaining the list serve and New Mexico website during past years. Mary Camire, Joy Winzerling, Ray Glahn, and Denis Medeiros will design outlines for web dissemination of information to be evaluated by committee members. A discussion of roster correction ensued since several members have not attended meetings for sometime. Tammy Bray will check with the appropriate Agricultural Experiment Station Directors for these individuals status in the group. Tammy Bray indicated that the W1002 project would be terminated next year. One suggested approach for writing the new project proposal was for each person to write a portion of the proposal and submit it to a group who would compile the work. It was thought that the proposal needs to be unique and outcome driven. Joy Winzerling and Connie Weaver said they would help. The meeting will be held in Washington DC next year (2007) in conjunction with Experimental Biology. Connie Weaver will organize next years meeting with Etta Saltos. Judy Driskell announced that she will be leaving the group at the termination of the project (September 30, 2007). We will miss Judy and wish her the very best. Station reports were given by attending members. Afterwards, the meeting was adjourned. Terry Shultz Secretary

W1002 SAES-422: Accomplishments

Short term outcomes: The individual and collaborative research efforts of the W1002 group listed below have provided new information on the requirements and metabolism of trace metals, vitamins and phytochemicals which have suggested and will suggest dietary modifications to ensure optimal health and which may reduced risk of diseases such as cancer, heart disease and birth defects.

Outputs: The major outputs have been peer reviewed publications and reports.

Activities:

University of Maine (Mary Ellen Camire): A USDA NRICGP equipment grant provided the Camire laboratory with a Hologic Discovery W bone densitometer that was installed in the fall of 2005. The equipment will be used in osteoporosis and obesity research. Specifically, Dr. Camire is seeking funds to examine the role of appetite-suppressing foods on weight loss in overweight women.

The study of the role of anthocyanin-rich fruits in reducing risks for Type 2 diabetes just ended. Twenty-five overweight persons with elevated fasting glucose levels completed the study; 9 people were assigned to the anthocyanin group and asked to consume two servings of anthocyanin-rich foods per day for three months. Subjects were provided with a list of fruits and servings sizes that should provide about 180 mg of anthocyanins. The control group was matched to the anthocyanin group for size, age and gender, and asked to follow their typical diet, but consume no more than three servings of foods containing anthocyanins per week. The final 3-month laboratory values have not yet been analyzed. Fasting serum glucose levels were not lower in the anthocyanin group at 2 months into the study, but insulin values were significantly higher in the control group. Future work will focus on the effect of anthocyanins and related compounds on digestion and postprandial glucose and insulin responses.

Subject screening has begun for a project on the treatment of Helicobacter pylori infections with cranberry supplements. Adults who test positive for the infection using a breath test will be randomized to a commercially-available cranberry supplement or to a placebo. Three capsules will be consumed daily, with repeat breath tests three and six weeks after the start of supplementation.

University of California-Berkeley (Barry Shane): Cellular, metabolic and kinetic effects of genetic polymorphisms on folate metabolism. The Shane group have continued studies on the metabolic and nutritional effects of common polymorphisms in human folate-related genes that have been shown to influence disease risk. They identified a gene-gene interaction that influences the risk of heart disease and are developing cell culture models to mimic this interaction. They have continued to evaluate two new genetic mouse models to mimic the effects of these polymorphisms and to evaluate their effects on metabolism and how this is modified by nutritional status. These new animal models have gene interruptions that disturb mitochondrial folate metabolism and histone methylation, a process involved in regulation of gene expression. They have continued to study one carbon metabolic fluxes and DNA methylation in these animals and in embryonic fibroblasts. Their data indicate that disturbances in mitochondrial folate metabolism result in early embryonic lethality.

In collaborative human studies with Jess Gregory at Florida, they have shown that a common polymorphism in the MTHFR gene causes elevated homocysteine prior to defective remethylation in folate-restricted young women. They have continued to evaluate genetic risk factors for neural tube defects.

Impact: Neural tube defects are the most common birth defects in humans and identification of genetic risk factors for this condition will allow screening to identify at risk individuals. Polymorphisms in genes encoding folate-dependent enzymes have been implicated as risk factors for cancer and vascular disease. The studies may indicate whether this risk can be modified by dietary changes.

Kansas State University (Denis M. Medeiros): Iron and bone work - Dr. Medeiros previously reported that severe iron deficiency negatively impacts bone microarchitecture. He has now determined whether marginal iron restriction that reflects some human consumption patterns could have similar consequences. Thirty-two weanling female, rats were randomly divided into four groups and fed the following diets for 10 weeks: 1) iron adequate, calcium adequate (FeA:CaA), 2) calcium restricted (FeA:CaR), 3) iron restricted (FeR:CaA), and 4) both calcium and iron restricted (FeD:CaR). DEXA analysis revealed that CaR decreased bone mineral density (BMD), and FeR decreased whole body bone mineral content (BMC). Iron restricted and calcium restricted groups had lower BMD than their adequate counterparts. All treatment restricted groups had lower BMD in the 4th lumbar (L-4) vertebrae than the FeA:CaA group. Vertebrae BMD was lower in all treatment groups compared to the control group and for BMC, the CaR groups were lower than the CaA groups and the FeR groups were lower that the FeA groups.and BMC were lower in iron and calcium restricted groups. The microarchitecture of the L-4 vertebrae was compromised in FeA:CaR, FeR:CaA, and FeR:CaR: 1) the connectivity density was reduced by FeR and by CaR; and 2) trabecular number was decreased and trabecular separation was increased by FeR ; Cortical thickness of the femur was reduced by both FeR and CaR. Finite element analysis revealed that L-4 vertebrae from the FeR:CaA group had greater internal stress with an applied force than the FeA:CaA group, thus would be more likely to break. Chelation of iron in cultured osteoblast cells impaired mineralization, but had no impact upon Type I collagen deposition. Iron depletion, similar to that occurring among some human populations reduced bone strength and microarchitecture based on in vivo and in vitro results reported here. Impaired mineralization with iron depletion appears to be a possible mechanism for the observed bone abnormalities.

Copper and cardiac mitochondrial biogenesis. The Medeiros laboratory continues to research the impact of copper-deficiency upon mitochondrial biogenesis as affected by copper deficiency. Using super-array technology, they conducted a study to determine whether certain metabolic and mitochondrial biogenic pathways were affected as a result of copper-deficiency in the heart. Rats were fed either control diets or diets deficient in copper from weaning until 5 weeks thereafter. RNA was obtained and subsequent hybridization to customized super-arrays were carried out. The genes spotted on the super-arrays included sequences that encode for connective tissue proteins, substrate utilization enzymes for fatty acid and glucose utilization, apoptosis proteins, and cell cycle genes. Early analysis suggests that NFkB-1, a key enzyme involved in cell proliferation and apoptosis was up-regulated in copper deficient rat hearts. Another study using rats suggested that marginal copper intakes at levels likely to be consumed by westerners, can lead to abnormal electrocardiograms in the absence of ultra-structural pathologies, but with enhanced lipid deposition in the heart.

Impact: Marginal iron intake has a negative consequence upon bone health. The levels used in the study are cause for concern in that many humans consume marginal iron diets worldwide. Iron may be an overlooked factor in bone health. The role of copper in maintaining the integrity of the heart may be related to NFkB, a critical gene in cell proliferation. Marginal copper intakes can lead to abnormal electrical activity of the heart.

University of Nebraska (Judy Driskell): Anthropometric measurements and selected nutrient intakes of Latino immigrant children, 4-8 y, in rural Nebraska were determined. 19.4% were at risk of being overweight and 11.1% were overweight. 72% had higher fat intakes; 94.4%, higher saturated fat intakes; and 33.3%, higher cholesterol intakes than recommended. These Latino immigrant children had higher median energy, protein, fat, folate, vitamin C, iron, and calcium intakes than reported for school children, 4-11 y, in a 1998 national Mexican Survey. Anthropometric measurements, selected nutrient intakes, and hemoglobin, hematocrit, and serum cholesterol values of 65 male and 66 female Koreans, 2-6 y, were determined. 8.4% were underweight; 10.7%, at risk of being overweight; and 4.6%, overweight. Hemoglobin concentrations of 3.8% were <110 g/L, 4.6% had hematocrits <.33, and 3.8% had serum cholesterol >5.17 mmol/L. The majority of subjects met the Korean Estimated Average Requirements and U.S./Canadian Estimated Average Requirements for protein, phosphorus, iron, zinc, and vitamin C. Blood concentrations of vitamin A, carotenoids, and vitamin E are currently being quantitated. Anthropometric measurements, selected nutrient intakes, and food beliefs have been collected from over 70 urban American Indians in the Lincoln, NE area. Selected vegetables were prepared by induction stir-frying and carotenoid concentrations and sensory attributes determined using a 78-member consumer panel of college students, 19-30 y. The cooked vegetables retained more than 85% of all their carotenoids (beta-carotene, alpha-carotene, beta-cryptoxanthin, lutein, lycopene). Over three-quarters of the panelists reported liking extremely/liking very much or liking moderately/liking slightly the appearance, color, flavor, texture, and overall acceptability of carrot, green onion, red pepper, snow pea, and yellow pepper; whereas, half to three-quarters reported similarly for white onion. Few differences in attribute responses were observed by gender. Induction stir-frying is a convenient method for cooking vegetables in which most of the carotenoids are retained, while still maintaining their acceptability/desirability to consumers.

Impact: Young children of Latino immigrants living in rural Nebraska have greater nutrient intakes than reported in a national Mexican survey, which is suggestive of acculturation-related dietary changes, some of which are undesirable. Young children in Korea appear to be in the acceptable weight category, have adequate dietary intakes, and have normal hemoglobin, hematocrit, and serum cholesterol values. Cooking vegetables by induction-stir-frying appears to be a desirable method in terms of cooking yield, carotenoid retention, and sensory characteristics.

Oregon State University (Emily Ho): The classic view of cancer etiology is that genetic alterations damage DNA structure and induce mutations resulting in non-functional proteins that lead to disease progression. More recently, the role of EPIGENETIC alterations during cancer has gained increasing attention. These epigenetic alterations affect gene expression without directly changing DNA sequences, but rather turn on or off gene expression by post-translational modifications. Interestingly, many of these epigenetic modifications can also be modified by dietary factors. For example, pharmacological histone deacetylase (HDAC) inhibitors are currently being tested in human clinical trials and are proposed to have potent anti-cancer activity. The Ho laboratory have found that sulforaphane, a chemical found in cruciferous vegetables is also an HDAC inhibitor, increases acetylated histone levels and has anti-cancer properties in the prostate. Addition of 15 mM SFN inhibited HDAC activity and induced accumulation of acetylated histones in both normal and prostate cancer cells. However, SFN induced expression of p21 and a G2/M arrest only in PC3 cells, with no effect in PrEC. Thus, similar to other HDAC inhibitors, SFN exhibited selective anti-proliferative effects to cancer cells. Secondly, they examined the effects of SFN on HDAC inhibition and prostate chemoprevention in vivo using a xenograft model. Dietary SFN (~7.5 mmol per day) caused a significant decrease in HDAC activity and increase in histone acetylation in the xenografts, prostates and peripheral blood mononuclear cells (PBMC) and suppressed the growth of PC3 xenografts. Thus, consumption of SFN in the diet was able to inhibit HDAC activity in circulating blood cells and prostate, and retard prostate tumor growth in vivo. These findings provide evidence that one potential mechanism by which SFN acts as a cancer chemopreventive agent is by the inhibition of HDAC activity.

Impact: Prostate cancer remains a very significant contributor to the morbidity and mortality of U.S. men. Increased vegetable intake has long been associated with decreases in prostate cancer risk. However, mechanisms for chemoprevention are unclear. This research is significant because nutritional strategies that target aberrant epigenetic alterations, such as acetylation, in prostate cancer have the potential to dramatically reduce the incidence of prostate cancer, reduce health care costs associated with prostate cancer, and improve the quality of life of thousands of American men. This work will form the basis for future work and larger trials to identify effective dietary intervention strategies that are broadly applicable nutrition recommendations and will significantly reduce the burden of prostate cancer. University of Connecticut (Sung Koo): Dr. Koo and associates have been studying whether there is a relationship between plasma cholesterol response and plasma lutein and vitamin E responses after egg consumption. Using a crossover design, postmenopausal women (50-68y) were assigned to a pasteurized whole egg treatment containing 640 mg cholesterol, 600 mcg lutein/zeaxanthin/d (EGG) or an egg substitute treatment containing 0 mg cholesterol, 0 mcg lutein/zeaxanthin and 540 mcg beta-carotene/d (SUB) for 30 d, followed by a 3 wk washout period and 30 d alternate treatment period. Nine women were classified as hyper-responders to cholesterol and 17 women were classified as hypo-responders to cholesterol. After the EGG period cholesterol hyper-responders had a significantly greater increase in plasma lutein (42%) and alpha-tocopherol (33%) compared to the cholesterol hypo-responders who has a 23% increase in lutein and no increase in alpha-tocopherol. Increase in plasma lutein from eggs did not affect other carotenoids in the blood. After the SUB period, the 63% increase in plasma beta-carotene observed in the cholesterol hyper-responders was significantly greater (p<0.05) than the 39% increase observed in the cholesterol hypo-responders. These results suggest plasma responses to dietary cholesterol, carotenoids and alpha-tocopherol are related. The Koo laboratory have also examined whether the inhibition of intestinal lipid absorption by green tea is associated with the inhibitory effect of its catechins on pancreatic phospholipase A2 (PLA2). PLA2 activity was assayed by using 1,2-dioleoylphosphatidylcholine (DOPC), porcine pancreatic PLA2, and catechins at varying concentrations (0.075-1.80 micromol/L). The amount of 1-oleoyl-2-hydroxyphosphatidylcholine liberated was determined by HPLC. The percentages of inhibition of PLA2 by (-)-epicatechin (EC), (+)-catechin (CAT), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), and (-)-epigallocatechin gallate (EGCG) at 0.6 micromol were 23.3, 24.8, 25.7, 39.7, and 64.9%, respectively. In an in vivo study, ovariectomized rats with lymph cannula were infused intraduodenally for 8 h with a triolein emulsion containing [dioleoyl-1-14C] PC, DOPC, alpha-tocopherol (TP), and retinol (ROH) without (CAT0) or with CAT or EGCG. The lymphatic total 14C-radioactivity (14C) was significantly lowered by EGCG (45.5% dose) compared with CAT (56.2% dose) and CAT0 (64.7% dose). The 14C radioactivity remaining in the small intestinal lumen and cecum was higher in EGCG (24.1% dose) than in CAT (9.5% dose) and CAT0 rats (9.0% dose). Significantly less 14C radioactivity was incorporated into lymph triacylglycerol and cholesteryl ester in EGCG rats. The absorption of TP, used as a marker of extremely hydrophobic lipids, was significantly lower in EGCG (7.8 micromol) than CAT (14.4 micromol) and CAT0 rats (16.8 micromol). The absorption of ROH was unaffected, whereas oleic acid output was lower in EGCG rats. The results show that EGCG inhibits the intestinal absorption of lipids, which is in part associated with its inhibition of PC hydrolysis. Impact: Study results show: 1) Egg may be used as a vehicle for delivering bioavailable carotenoids in humans and 2) Green tea or catechins lower the intestinal absorption of fat and fat-soluble compounds by inhibiting pancreatic phospholipase A2. The findings indicate that the absorption and bioavailability of fat-soluble compounds are significantly influenced by egg intake in humans and green tea in rats. Data may be used as a basis for specific dietary recommendations regarding the inclusion of eggs and green tea in diets for the prevention and treatment of certain diet-related diseases and disorders.

Oklahoma State University (Barbara J. Stoecker): The Stoecker laboratory have collaborated with several investigators to examine effects of nutrients and phytonutrients on bone strength and structure. Highlights of three projects completed are included below:

Effects of marginal iron status on microarchitecture and strength of bone This study was a collaboration with Dr. Dennis Medeiros and is reported in detail in the Kansas State University W-1002 report. In summary, iron restriction produced a number of changes in microarchitecture in addition to those seen with calcium restriction alone. Connectivity density was decreased both by iron and by calcium restriction. Trabecular number was decreased and trabecular separation was increased by iron restriction. When a force was simulated on the 4th lumbar vertebra, bones from animals fed calcium adequate but iron restricted diets showed greater stress than bones from animals fed calcium adequate, iron adequate diets.

Effects of enhanced antioxidant status on bone quality in a rat model of male osteoporosis One-year-old retired breeder rats were randomly assigned to orchidectomy (ORX) or sham-surgery (SHAM) treatments. Three days post-surgery, ORX rats were assigned to the following treatments: ORX, ORX + 2.5% powdered orange pulp, ORX + 5.0% orange pulp and ORX + 10% orange pulp. This isonitrogenous, isocaloric modified AIN-93M diet was fed for four months with all ORX rats pair fed to the mean intake of the sham group. At the termination of the study, blood and bone samples were collected. Plasma antioxidant capacity was determined spectrophotometrically and bone microarchitecture was assessed using microcomputed tomography (MicroCT-40, Scanco Medical). Data were analyzed using the General Linear Model procedure in SAS (version 9.1) to determine the effects of orange pulp on plasma total antioxidant capacity and on bone microarchitecture. Lack of testosterone decreased (p<0.05) antioxidant status and even the 2.5% level of orange pulp increased antioxidant capacity of ORX rats to that of the SHAM animals. In trabecular cores from the 4th lumbar vertebrae, ORX rats had significantly reduced bone volume fraction, connectivity density, and trabecular number and increased trabecular separation. In ORX rats fed 10% powdered orange pulp, bone volume fraction, and trabecular number were significantly increased and trabecular separation significantly decreased compared to ORX animals. This study was a collaboration with Dr. Farzad Deyhim at Texas A&M University-Kingsville and Dr. B.S. Patil at Texas A&M UniversityCollege Station.

Effects of high does of vitamin E on bone quality in a rat model of male osteoporosis Vitamin E is commonly known to function as an antioxidant. However, little research has been done to discover the effects that Vitamin E might have on bone quality. In this experiment, thirty-two rats were randomly assigned to four treatment groups. Eight rats were sham operated and 24 were orchidectomized. The groups were: Control (SHAM), orchidectomy (ORX), low dose supplemental vitamin E of 65.6 mg/kg diet (ORX+LD), or high dose vitamin E of 656 mg/kg diet (ORX+HD). After four months of feeding, microarchitecture of the 4th lumbar vertebra (L4) were analyzed using a µCT 40 scanner. Orchidectomy significantly reduced bone volume fraction, trabecular number and connectivity density, and increased trabecular separation. Microarchitecture of the ORX+LD group did not differ significantly from the ORX group. Bone volume fraction of the ORX+HD group tended to be lower than the ORX group (15.4 ± 0.6 vs 18.5 ± 1.3, p=0.11) and trabecular thickness was significantly reduced in the ORX+HD group. Bone strength was estimated using finite element analyses. Compared to SHAM, the ORX group required significantly less force for compression and demonstrated decreased stiffness and size independent stiffness and increased von Mises stresses. Strength predictions for the ORX+LD dose did not differ from the ORX group. The ORX+HD showed significantly increased von Mises stresses compared to the ORX group. The changes in bone with ORX demonstrate the importance of testosterone or its metabolites in maintenance of bone quality. The high dose Vitamin E (ORX+HD) had detrimental effects on the microarchitecture and predicted stress with load of the L4 vertebra, suggesting reduced bone quality. This study was a collaboration with Dr. Farzad Deyim at Texas A&M University-Kingsville.

Impact: Various dietary components have the potential to change bone strength and microarchitecture and recent improvements in instrumentation have increased our capacity to measure these effects. Dietary components that affect antioxidant status may be beneficial or detrimental to bone quality depending on dose. Many people take vitamin E supplements greater than 10-fold the Recommended Dietary Allowance and assume these levels of supplements are safe. However, our results raise the concern that high dose vitamin E supplements may have detrimental effects on bone; further studies addressing mechanisms for these effects on microarchitecture and studies in humans are needed. The effects of adding orange pulp to animal diet were positive at the dose levels tested; studies are underway to determine if specific components of orange pulp were responsible for the beneficial effects on bone microarchitecture. Finally, the work with marginal iron intake is important because many women and children around the world are deficient in iron. To date, little attention has been given to possible effects of iron status on development of optimal bone mass.

University of Arizona (Joy Winzerling): Dr. Winzerling was unable to continue with the original objectives proposed for her regional research project because her collaborator was in a severe accident. There are several researchers in her group working on smoking in relationship to disease. Her laboratory is now contributing in the area of human lung cell iron metabolism and comparative iron metabolism in disease insect vectors. Inhalation of airborne pollution particles that contain iron can result in a variety of detrimental changes to lung cells and tissues. A primary concern is the potential for iron-mediated oxidative stress. The lung iron burden can be substantially increased by exposure to cigarette smoke and smoke contains environmental toxins that could influence intracellular iron status. Previously, she reported that although cigarette smoke contains iron particles that enter lung cells, the exposure of lung cells to smoke down regulates ferritin synthesis. Ferritin is important because it is the primary protein for storing iron and decreased levels increase the potential for iron-mediated oxidative stress. Her findings suggest that this mechanism could be compromised in lung cells at a time when the oxidative stress potential is increased. She now reports that iron also alters levels of protein called early growth response 1. This protein controls the synthesis of proteins involved in cell division. Message levels of EGR1 determine how much protein is made. However, in lung cells iron uncouples EGR1 protein levels from EGR1 message levels. Message levels are sustained with iron exposure time, while protein levels decline. She does not yet know the implications of this finding. However, if Egr1 is needed as part of the process for the replacement of cells damaged by smoke, then iron in smoke particulate matter could possibly effect cell replacement. Dr. Winzerlings work in mosquitoes is directed toward evaluating the role of ferritin in these animals. Female mosquitos blood feed to complete their lifecycle. The blood meal is high in iron and females have developed mechanisms that allow use of the iron load in egg development while avoiding oxidative stress. She has found that the majority of iron from a blood meal is moved to the ovaries for egg development indicating a crucial role for iron in this process. She also has found that mosquitoes secrete ferritin in response to the iron load by a non-classical pathway. These findings contrast with those for mammals where ferritin for iron storage is found in the cell cytoplasm.

Impact: Others in the group are working in the area of smoking and nutrition. Our data complements their work. The findings of the lung iron project are important in the short term because they provide insight into the mechanisms that could promote lung cancer, a primary form of cancer in the US. Future work in this area could lead to dietary recommendations for iron intake for smokers or for individuals exposed routinely to smoke that differ from the general population. The mosquito project is important in the short term because iron is required for egg development. Mosquitoes transmit hundreds of millions of cases of disease each year because of the large numbers of these disease vectors. If the iron pathway differs from that of mammals, we can identify potential targets for mosquito control that could reduced the numbers of disease vectors. Purdue University (Connie Weaver): The Weaver laboratory has been studying the effect of isoflavones on calcium metabolism and bone. They completed their dose response study of the effect of soy isoflavones in a soy protein powder on 44Ca absorption and postmenopausal women using novel 41Ca technology. Thirteen postmenopausal women (>6 years since menopause) were pre-dosed with 41Ca intravenously. After a 100-day baseline period, subjects were given 40 grams of soy protein per day that contained either 0, 97.5, or 135.5 mg of total isoflavones in randomized order. The soy protein isolate powder was incorporated into baked products and beverages. Each 50-day treatment phase was followed by a 50-day recovery phase. Serum isoflavone levels, biochemical markers, and Ca absorption were measured at the end of each phase. Twenty-four hour urine collections were performed every ~ 10 days during each phase for 41Ca/Ca analysis by Accelerator Mass Spectrometry. Serum isoflavone levels reflected the mount of isoflavones consumed in a dose¬-dependent manner. None of the isoflavone levels had a significant effect on biochemical markers of bone turnover, urinary cross-linked N-teleopeptides of type I collagen (NTx) and serum osteocalcin (OC), 44Ca absorption or bone turnover as assessed by urinary 41Ca/Ca. In conclusion, soy isoflavones up to 135.5 mg per day did not suppress bone resorption in postmenopausal women. She now has data showing that bone resorption was significantly reduced by estrogen and a bisphosphonate. The Weaver group have also carried out calcium bioavailability studies. They conducted acute and chronic feeding studies on the effect of purported calcium absorption enhancing components on calcium absorption and retention and bone measures in growing rats. Whey protein and honey and its constituents enhanced calcium absorption in a dose response manner in single meal experiments, but the effect largely disappeared on chronic feeding attributable to adaptation through the calcium homeostatic mechanism.

Washington State University (Terry D. Shultz): Hypertension is a major controllable risk factor associated with cardiovascular disease events, such as myocardial infarction, stroke, and heart failure. Cleavage of angiotensinogen by rennin produces angiotensin I, which is subsequently hydrolyzed by angiotensin-I-converting enzyme (ACE) to angiotensin II (a potent vasoconstrictor). It has been reported that in vitro incubation of milk proteins with proteinase preparations can result in the release of ACE inhibitory peptides. The Shultz group examined three commercial proteases, Validase BNP L, Validase FP II, and Multifect Neutral for digestion of 20% solutions of whey protein concentrate 80 (WPC80) for production of ACE inhibitory activity. Enzymes were used at 1% (w/w) and incubated at pH 7 for up to 200 min at 50 C. Degree of hydrolysis (DH) was monitored by TNBS assay and ACE inhibition measured using hippuryl-L-histidyl-L-leucine as substrate. Incubation for 5-15 min produced DH of 0.5-3% and maximal ACE inhibitory activity. Validase FP II produced 61% inhibition with an IC50 value of 4.1 mg/mL, Validase BNP L 46% inhibition with an IC50 value of 5.7 mg/mL, and Multifect Neutral 32% with an IC50 value of 8.1 mg/mL. The Validase BNP L hydrolysate was selected for nutraceutical beverage development because the enzyme is readily inactivated by lowering pH.

Impact: This research brings novel application to the dairy industry: utilization of whey proteins in a beverage with potential use as a dietary treatment for hypertension.

Milestones: Peer review articles have been published and a number of new extramural research grants have been obtained. New methods for assessing bone function and for measuring trace metal and vitamin turnover have been developed which have allowed a number of new collaborative projects within the W1002 group.

Journal Articles - Peer Reviewed

Braun, M. and Weaver, C.M. A call to evaluate the impact of calcium-fortified foods and beverages. Nutr. Today 41 :40-47,2006.

Britten, P., Lyon, J., Weaver, C.M., Kris-Etherton, P., Nicklas, T., Weber, J., Davis, C. MyPyramid food intake pattern modeling for the Dietary Guidelines Advisory Committee. JNEB In press, 2006.

Cai, D.J., Zhao, Y., Glasier, J., Cullen, D., Barnes, S., Turner, C.H., Wastney, M., Weaver, C. Comparative effect of soy protein, soy isoflavones and 1713-estradiol on bone metabolism in adult ovariectomized rats. J Bone Miner Res, 20:828-39, 2005.

Camire, M.E., Dougherty, M.P. 2005. Internet survey of nutrition claim knowledge. J Food Sci. Education 4(2):18-21.

Camire ME, Dougherty MP and Briggs JL. 2005. Antioxidant-rich foods retard lipid oxidation in extruded corn. Cereal Chemistry. 82(6):666-670.

Clark, R.M., Herron, K.L., Waters, D., and Fermandez M.L. (2006) Low and High Plasma Response to Egg Cholesterol Predicts Response to Lutein and Beta-Carotene in Men and Women. J. Nutr. (accepted Dec. 23, 2005).

Dashwood, RH, Myzak, MC and Ho, E. (2005) Dietary HDAC inhibitors: time to rethink weak ligands in cancer chemoprevention? Carcinogenesis. 2006 Feb;27(2):344-9

Davis, S. R., Quinlivan, E. P., Shelnutt, K. P., Ghandour, H., Capdevila, A., Coats, B. S., Wagner, C., Shane, B., Selhub, J., Bailey, L. B., Shuster, J. J., Stacpoole, P. W. and Gregory III, J. F. (2005). Homocysteine is elevated but total remethylation is unchanged by the methylenetetrahydrofolate reductase 677C®T polymorphism and by dietary folate restriction in young women. J. Nutr. 135:1045-1050.

Dayal, S., Devlin, A. M., McCaw, R. B., Liu, M.-L., Arning, E., Bottiglieri, T., Shane, B., Faraci, F. M. and Lentz, S. R. (2005). Cerebral vascular dysfunction in methionine synthase-deficient mice. Circulation 112:737-744.

Deyhim F, Stoecker BJ, Brusewitz GH, Devareddy L, Arjmandi BJ. (2006) Dried plum reverses bone loss in an osteopenic rat model of osteoporosis. Menopause (In Press)

Driskell JA, Kim Y-N, Goebel KJ. Few differences found in the typical eating and physical activity habits of lower-level and upper-level university students. J Am Diet Assoc 2005; 105:798-801.

Driskell JA, Marchello MJ, Giraud DW, Sulaeman A. Vitamin and selenium content of ribeye cuts from grass- and grain-finished bison of the same herd. J Food Qual 2004; 27:388-398.

DSouza, N., Skonberg, D.I., Camire, M.E., Guthrie, K.E., Malison, J., Lim,a L. 2005. Influence of dietary genistein levels on tissue genistein deposition and on the physical, chemical, and sensory quality of rainbow trout, Oncorhynchus mykiss. J. Agric. Food Chem. 53(9): 3631-3636.

Gailer J, George GM, Pickering IE, Prince RC, Younis HS, Winzerling JJ. (2002) Biliary excretion of [(GS)(2)AsSe](-) after intravenous injection of rabbits with arsenite and selenate. Chemical Research in Toxicology. 15:1466-71.

Geiser DL, Chavez CA, Flores-Munguia R, Winzerling JJ, Pham DQ. Aedes aegypti ferritin. Eur J Biochem. 2003 Sep;270(18):3667-74.

Geiser DL, Zhang D, Winzerling JJ. Secreted ferritin: Mosquito defense against iron overload? Insect Biochem Mol Biol. 2006 Mar;36(3):177-87.

Gunther, C.W., Legowski, PA, McCabe, L.D., McCabe, G.P., Peacock, M., Lyle, R.M., Weaver, C.M., Teegarden, D. Parathyroid hormone is associated with decreased fat mass in young health women. Int. J. Obesity. 30:94-99, 2006.

Hansen, C.M., Shultz, T.D., Hunt, K.C., Hardin, K., Leklem, J.E., Huang, A. & Ames, B.N. (2006) Vitamin B-6 intake and smoking status influence lymphocyte serine hydroxymethyltransferase activity in healthy adults. Submitted.

Hansen, C.M., Shultz, T.D., James, S.J., Melnyk, S., Leklem, J.E., Hardin, K. & Ames, B.N. (2006) Effects of smoking and vitamin B-6 intake on plasma thiol concentrations. Preparation.

Hansen, C.M., Shultz, T.D., James, S.J., Melnyk, S., Leklem, J.E., Hardin, K. & Ames, B.N. (2006) Effects of smoking and vitamin B-6 intake on plasma thiol concentrations. Preparation.

Hansen, C.M., Shultz, T.D., Hunt, K.C., Hardin, K., Leklem, J.E., Huang, A. & Ames, B.N. (2006) Vitamin B-6 intake and smoking status influence lymphocyte serine hydroxymethyltransferase activity in healthy adults. Submitted.

Heaney, RP. and Weaver, C.M. Newer perspectives on calcium and bone quality. J. Am. Coil. Nutr 24(6):574S-581S, 2005.

Kling, P.J. and Joy J. Winzerling (2002) Iron status and the treatment of the anemia of prematurity. Clinics in Perinatology 29(2):283-94.

Lim, U., Peng, K., Shane, B., Stover, P. J., Litonjua, A. A., Weiss, S. T., Gaziano, J. M., Strawderman, R. L., Raiszadeh, F., Selhub, J., Tucker, K. L. Cassano, P. A. (2005). Polymorphisms in cytoplasmic serine hydroxymethyltransferase and methylene-tetrahydrofolate reductase affect the risk of cardiovascular disease in Men. J. Nutr. 135:1989-1994.

Mashiyama, S.T., Hansen, C.M., Roitman, E., Sarmiento, S., Leklem, J.E., Shultz, T.D. & Ames, B.N. (2006) DNA-uracil in healthy humans measured with an improved assay (uracil assay V3.0) is not affected by marginal vitamin B-6 deficiency. Submitted.

Mayo JJ, Kohlhepp P, Zhang D, Winzerling JJ. Effects of sham air and cigarette smoke on A549 lung cells: implications foriron-mediated oxidative damage. Am J Physiol Lung Cell Mol Physiol. 2004 Apr;286(4):L866-76

Medeiros DM, Shiry LJ, McCune SA. 2005. Marginal copper intakes over a protracted time period in genetically and non-genetically susceptible heart disease rats disturbs electrocardiograms and enhances lipid deposition. Nutr. Res. 25: 663-672.

Myzak, MC, Hardin, K. Dashwood, RH and Ho, E. (2004) Sulforaphane Inhibits Histone Deacetylase Activity in BPH-1, LnCaP, and PC-3 Prostate Epithelial Cells. Cancer Epidemiology biomarkers and prevention, 13:1847S.

Myzak, MC, Hardin, K, Rong W, Dashwood, RH and Ho, E. (2005) Sulforaphane inhibits histone deacetylase activity in BPH-1, LnCaP, and PC-3 prostate epithelial cells. Carcinogenesis. 2005 Nov 15; [Epub ahead of print]

Nichol, Helen, John H. Law and Joy J. Winzerling. (2002) Iron Metabolism in Insects. Annual Review of Entomology, 47:535-559.

Nichol H, Winzerling, JJ (2002) Structured RNA upstream of insect cap distal iron responsive elements enhances iron regulatory protein-mediated control of translation. Insect Biochemistry and Molecular Biology, 2002 32(12):1699-710.

Nicklas, T.A., Weaver, C.M., Stitzel, K.F. The 2005 Dietary Guidelines Advisory Committee: Developing a Key Message. JADA 105(9):1418-24,2005.

OLeary, V. B., Mills, J. L., Pangilinan, F., Kirke, P. N., Cox, C., Conley, M., Weiler, A., Peng, K., Shane, B., Scott, J. M., Parle-McDermott, A., Molloy, A. M., Brody, L. C., Members of the Birth Defects Research Group. (2005). Analysis of methionine synthase reductase polymorphisms for neural tube defects risk association. Molec. Genet. Metab. 85:220-227.

Parelman M, Stoecker B, Baker A, Medeiros DM. 2006. Iron restriction negatively affects bone in female rats and mineralization of hFOB osteoblast cells. Exp. Biol. Med. (Maywood) In Press.

Parelman M, Stoecker B, Baker A, Medeiros DM. (2006) Iron restriction negatively affects bone in female rats and mineralization of hFOB osteoblast cells. Exp Biol Med (In Press).

Pham DQ, Kos PJ, Mayo JJ, Winzerling JJ. Regulation of the ribonucleotide reductase small subunit (R2) in the yellow fever mosquito, Aedes aegypti. Gene. 2006 Mar 9;

Pham Q.D. D, Bluchuta B,Nichol H,Winzerling JJ. (2002) Cloning, sequencing and expression of RNR R1 and R2 in mosquito. Insect Biochemistry and Molecular Biology, 32(9):1037-44.

Quinlivan, E. P., Davis, S. R., Shelnutt, K. P., Henderson, G. N., Ghandour, H., Shane, B., Selhub, J., Bailey, L. B., Stacpoole, P. W. and Gregory III, J. F. (2005). Methylenetetrahydrofolate reductase 677C->T polymorphism and folate status affect one-carbon incorporation into human DNA deoxynucleosides. J. Nutr. 135:389-396.

Reinwald, S. and Weaver, C.M. Soy isoflavones and bone health: are they a double¬ edged sword? J. Natural Products In press, 2006.

Ritzenthaler, K.L., McGuire, M.K., Shultz, T.D., McGuire, M.A., Koepp, A.E., Luedecke, L.O., Hanson, T.W., Dasgupta, N. & Chew, B.P. (2005) Consumption of conjugated linoleic acid (CLA) from CLA-enriched cheese does not alter milk fat or immunity in lactating women. J.Nutr. 135:422-430.

Ritzenthaler, K.L., Shultz, T.D., McGuire, M.A., Shahin, M.A., Dasgupta, N. & McGuire, M.K. (2006) Concentrations of conjugated linoleic acid (CLA) in human plasma and lipid fractions and their relationship with dietary CLA intake. Preparation.

Ritzenthaler, K.L., Shultz, T.D., McGuire, M.A., Shahin, M.A., Dasgupta, N. & McGuire, M.K. (2006) Concentrations of conjugated linoleic acid (CLA) in human plasma and lipid fractions and their relationship with dietary CLA intake. Preparation.

Shultz, T.D., Hansen, C.M., Hardin, K., Leklem, J.E. & Ames, B.N. (2006) Assessment of vitamin B-6 status in young men and women cigarette smokers and nonsmokers consuming a controlled diet. Submitted.

Smith BJ, Lucas EA, Turner RT, Evans GL, Lerner MR, Brackett DJ, Stoecker BJ, Arjmandi BH. Vitamin E provides protection for bone in mature hindlimb unloaded male rats. (2005) Calcified Tissue Int 76:272-279.

Spence, L.A, Lipscomb, E.R, Cadogan, J., Martin, B., Wastney, ME, Peacock, M., Weaver, C.M. The effect of soy protein and soy isoflavones on calcium metabolism and renal handling in postmenopausal women: A randomized cross over study. Am. J. Clin. Nutr. 81 :916-922, 2005.

Stempak, J. M., Sohn, K.-J., Chiang, E.-P., Shane, B. and Kim, Y.-I. (2005). Cell and stage of transformation-specific effects of folate deficiency on methionine cycle intermediates and DNA methylation in an in vitro model. Carcinogenesis 26:981-990.

Teh, Y.-H., Dougherty, M.P., and Camire, M.E. 2005. Frozen blueberry-soy dessert quality. J. Food Sci. 70(2):S119-22.

Weaver, C.M. Inulin, oligofructose and bone health: experimental approaches and mechanisms. Br. J. Nutr. 93(1 ):S99-S1 03, 2005.

Weaver, C.M. and Cheong, J.MK Soy isoflavones and bone health: The relationship is still unclear. J. Nutr. 135:1243-1247,2005

Welch, J.M. and Weaver, C.M. Calcium and exercise affect the growing skeleton. Nutr. Reviews 63(11):361-373, 2005.

Weaver, C., Nicklas, T., Britten, P. The 2005 Dietary Guidelines Advisory Committee Report. Nutr. Today 40:102-107,2005.

Wigertz, K., Palacios, C., Jackman, L.A, Martin, B.R, McCabe, L.D., McCabe, G.P., Peacock, M., Pratt, J.H., Weaver, C.M. Racial differences in calcium retention in response to dietary salt in adolescent girls. Am. J. Clin. Nutr. 81 :845-850,2005.

Winzerling JJ, Pham DQ. Iron metabolism in insect disease vectors: Mining the Anopheles gambiae translated protein database. Insect Biochem Mol Biol. 2006 Apr;36(4):310-21.

Zhang D, Dimopoulos G, Wolf A, Minana B, Kafatos FC, Winzerling JJ. (2002) Molecular characterization of two mosquito iron regulatory proteins. Insect Biochemistry and Molecular Biology, 32(5):579-89.

Zhao, Y., Martin, B.R., Weaver, C.M. Calcium bioavailability of calcium carbonate fortified soymilk is equivalent to cow's milk in young women. J Nutr. 135:2379-2392, 2005.

Zhao, Y., Martin, B.R, Wastney, M.E., Schollum, L., Weaver, C.M. Acute Versus Chronic Effects of Whey Proteins on Calcium Absorption in Growing Rats. Exp. BioI. Med. 230:536-542, 2005.

Extension Publication

Driskell JA. Nutrient Recommendations of Adults. NebGuide G-05-1555-A. 2005. Available at http://www.ianrpubs.unl.edu/epublic/pages/publicationD.jsp?publicationId=294.

Books/Book Chapters

Bagley, P. and Shane, B. (2005). Folate. In Encyclopedia of Dietary Supplements, Coates, P., Blackman, M. R., Cragg, G., Levine, M., Moss, J. and White, J., eds., pp. 219-228, Marcel Dekker, New York.

Heaney, RP. and Weaver, C.M. Ch. 7 Requirements for What Endpoint. In: Calcium in Human Health. Weaver, C.M. and Heaney, RP., eds. Humana Press. 97-104,2006.

Wastney, ME, Y. Zhao, Weaver, C.M. Ch. 6 Kinetic Studies. In: Calcium in Human Health. Weaver, C.M. and Heaney, RP., eds. Humana Press. 83-93, 2006.

Weaver, C.M. Ch. 17 Pre-Puberty and Adolescence. In: Calcium in Human Health. Weaver, C.M. and Heaney, RP., eds. Humana Press. 281-296,2006.

Weaver, C.M. Ch. 5 Clinical Approaches for Studying Calcium Metabolism and Its Relationship to Disease. In: Calcium in Human Health. Weaver, C.M. and Heaney, RP., eds. Humana Press. Pp 65-81, 2006.

Weaver, C.M. and Heaney, RP. Ch.9 Calcium In: Modern Nutrition in Health Disease. 10th Ed. M.E. Shils, M. Shike, AC. Ross, B. Caballero, RJ. Cousins, eds. Lippincott Williams & Wilkins Pp 194-210, 2006.

Weaver, C.M. and Heaney, RP., editors. Calcium in Human Health, Humana Press. Totowa, New Jersey 2006.

Weaver, C.M. and Heaney, RP. Ch. 9 Food Sources, Supplements and Bioavailability. In: Calcium in Human Health. Weaver, C.M. and Heaney, RP., eds. Humana Press. 129-142, 2006.

Winzerling, Joy J, and Daphne Q.-D. Pham (2004) Ferritin. In Comprehensive Insect Physiology, Biochemistry, Pharmacology and Molecular Biology. Biochemistry and Molecular Biology, Ed: Lawrence I. Gilbert,Kostas Iatrou,and Sarjeet Gill. Elsevier.

Winzerling, Joy J, and Daphne Q.-D. Pham (2004) Ferritin. In Comprehensive Insect Physiology, Biochemistry, Pharmacology and Molecular Biology. Biochemistry and Molecular Biology, Ed: Lawrence I. Gilbert,Kostas Iatrou,and Sarjeet Gill. Elsevier.

Theses/Dissertations Directed

Aldaous, S.A. 2005. Development Of Extruded Wild Blueberry Confection. M.S. Thesis, University of Maine, Orono, ME.

Michelle Braun. Bone mineral metabolism in adolescents, as assessed by metabolic balance studies, Ph.D. thesis, Purdue University

Jennifer Cheong. Effects of phytochemicals on bone turnover & validation of a novel method to assess bone turnover, Ph.D. thesis, Perdue University

Yong Jiang. Phytochemicals and bone, M.S. thesis, Purdue University

Lora KR. Vitamin B-6 status of a group of 4-8 y old children of Latino immigrants in rural Nebraska. M.S. Thesis. University of Nebraska-Lincoln. December, 2004.

Nunn MD. The effects of various cooking methods on the sensory qualities and carotenoid retention in selected vegetables. M.S. Thesis: University of Nebraska-Lincoln. December, 2004.

Oberholtzer, A.S. 2005. Development and Evaluation of a New Salmon Jerky Snack Product. M.S. Thesis, University of Maine, Orono, ME.

Parelman M. 2005. Marginal iron restriction negatively impacts bone. MS Thesis, Kansas State University

Yongdong Zhao. Calcium bioavailability of functional foods and discovery of bone biomarkers using serum proteomic approach, Ph.D. thesis, Purdue University

Abstracts

Lora KR, Giraud DW, Davy SR, Driskell JA. Vitamin B-6 status of 4-8 year old Latino children in rural Nebraska. FASEB J 2005;19(4):A433-434.

Driskell JA, Giraud DW, Davy SR, Lora KR. Vitamin E, vitamin C, and carotenoid status of children, 4-8 y, of Latino immigrants. FASEB J 2005;19(4):A475.

Presentations

Bailey B, Deyhim F, Stoecker BJ. Effects of vitamin E supplementation on bone quality. Experimental Biology 06, San Francisco, 2006.

Blake, P., Mayo, J.J., Kohlhepp, P. and Winzerling, J.J. (2003) Screening a Genomic Library for Aedes aegypti Ribonucleotide Reductase-R1 Subunit. Center for Insect Science University of Arizona, Tucson, AZ.

Camire, M.E. Anthocyanins: are they the fountain of youth? Kemin Foods, Des Moines IA. Oct. 5, 2005.

Camire, M.E., Guthrie, K., Bolton, J., Jordan, J.J., Kelley, S., Oberholtzer, A., Qiu, X., and Dougherty, M.P. Does color affect consumer attitudes about whole wheat baked products? AACC International Annual Meeting, Sept. 11-14, 2005, Orlando, FL.

Camire, M.E. Healthy Foods Laboratory Research. Wild Blueberry Health Summit, August 11-12 2005, Bar Harbor, ME. Camire, M.E., Dougherty, M.P., and The, Y.H. Frozen wild blueberry-soy desserts. Institute of Food Technologists Annual Meeting, July 16-20, 2005, New Orleans, LA.

Camire, M.E., Potter, R.M., Aldaous, S., Halteman, W., and Dougherty, M.P. Characteristics and sensory quality of wild blueberry soy beverages. Institute of Food Technologists Annual Meeting, July 16-20, 2005, New Orleans, LA.

Camire, M.E. Supplements for Men's Health. Togus Veterans Hospital, Augusta, ME. June 24, 2005.

Camire, M.E. Understanding the role of functional and nutraceutical food ingredients and the impact on anti-obesity formulations. 2nd Marcus Evans Ingredients-Based Obesity Symposium. Schaumburg, IL. June 8, 2005.

Camire, M.E. Complementary Medicine: Research and Education Challenges for the Nutrition Professional. Maine Nutrition Council Quarterly Meeting. Augusta, January 26, 2005.

Daniel, H. (presented by Camire, M.E.). Assessing the market for a blueberry and soy beverage. AACC International Annual Meeting, Sept. 11-14, 2005, Orlando, FL.

Garcia-Bañuelos, M., Luz Vazquez-Moreno1, Alfonso A. Gardea and Joy Winzerling. (2005) Differential Expression in Dormancy and Release of Dormancy in Apple (Malus domestica) shoots. American Society for Biochemistry and Molecular Biology. San Diego, CA.

Geiser, D.L., Jonathan J. Mayo, Meng-Chieh Shen and Joy J. Winzerling. (2005) Pilot Study: The Regulation by Iron of a Putative Divalent Metal Transporter in Anopheles gambiae Larval Cells, MOS55. American Society for Biochemistry and Molecular Biology. San Diego, CA. Student Travel Award Presentation.

Geiser, D. L., Jonathan J. Mayo, Meng-Chieh Shen and Joy J. Winzerling. (2005) Pilot Study: The Regulation by Iron of a Putative Divalent Metal Transporter in Anopheles gambiae Larval Cells, MOS55. Southwestern and Rocky Mountain Region of AAAS. Tucson, AZ.

Geiser, D., Mayo, J, Zhang, D., Kohlhepp, P. and Winzerling, J.J. (2002) Response of Aedes aegypti Larval Cells To Iron. Center for Insect Science Ninth Annual Poster Hexapodium. Tucson, AZ.

Geiser, D., Mayo, J, Zhang, D., Kohlhepp, P. and Winzerling, J.J (2002) Response of Aedes aegypti Larval Cells To Iron. Fourth International Symposium on Molecular Insect Science. Tucson, AZ.

Geiser, DL, Chavez, C, Pham, D. Q-D., Zhang, D., and Winzerling J.J. (2003) Response of Aedes aegypti Larval Cells To Iron. American Society of Biochemistry and Molecular Biology Annual Meeting. San Diego, CA

Mayo, J.J., Kohlhepp, P., Goodman, N. and Winzerling, J.J. (2003) Cigarette Smoke Influences Intracellular Iron Metabolism in A549 Lung Cells. College of Agriculture and Life Sciences, University of Arizona, Tucson, AZ.

Mayo, J.M.,,Kos, P., Shaffer, J., Pham, D.Q.-D. and Winzerling J. J. (2004) Characterization of Ribonucleotide Reductase in the Mosquito Aedes aegypti. 8th Annual American Society for Biochemistry and Molecular Biology Conference, Boston, MA (2004).

Mayo, J. J., Meng-Chieh Shen, Dawn L. Geiser and Joy J. Winzerling. (2005) Localization and Secretion of Ferritin in Cultured Aedes aegypti Larval Cells.: American Society for Biochemistry and Molecular Biology. San Diego, CA. Student Travel Award Presentation

Mayo, J. J., Meng-Chieh Shen, Dawn L. Geiser and Joy J. Winzerling. (2005) Localization and Secretion of Ferritin in Cultured Aedes aegypti Larval Cells. Southwestern and Rocky Mountain Region of AAAS. Tucson, AZ.

Meng-Chieh Shen, Jonathan J. Mayo, Dawn L. Geiser and Joy J. Winzerling. (2005) Localization and Secretion of Ferritin in Cultured Aedes aegypti Larval Cells . BIO5: Research Frontiers. The University of Arizona, Tucson, AZ 85721.

Morrow R, Deyhim F, Patil BS, Stoecker BJ. Feeding orange pulp improves bone microarchitecture in orchidectomized rats. Experimental Biology 06, San Francisco, 2006.

Parelman M, Stoecker B, Baker A, Medeiros DM. Marginal iron deficiency negatively impacts bone. Experimental Biology, San Diego, CA, 2005

Ramos-Clamont, G., Roberto Guzman Z. Joy Winzerling and Luz Vázquez-Moreno. (2005) Escherichia coli K88 Interaction with Serum Porcine IgA Oligosaccharides. Affinity 2005. Uppsala, Sweden. Young Investigators Award of Excellence.

Rickleffs K, Klaahsen DL, Medeiros DM. Copper deficiency and mitochondrial biogenesis: the role of transcriptional factors PPAR-alpha, PGC1-alpha, and NFKappaB1. Experimental Biology, San Francisco, 2005

Shane, B. Folate metabolism and retention: effect of dosage, Ortho-McNeil Pharmaceuticals, New Jersey, December 2004

Shane, B. Models for phenotypic characterization of nutrient  gene polymorphism interactions: studies on the euchromatic histone methyltransferase, methionine synthase and MTHFR mouse knock-outs, Fred Hutchinson Cancer Research Center, Seattle, Washington, April 2005

Shane, B. Gowland Hopkins Lecture - Molecular-genetic models for investigating the regulation of one-carbon metabolism, Thirteenth International Symposium on Chemistry and Biology of Folates and Pteridines, Egmond aan Zee, Netherlands, June 2005

Shane, B. Folate and vitamin B12 metabolism, Technical consultation on folate and vitamin B12, Department of Nutrition for Health and Development, WHO, Geneva, Switzerland, October 2005.

Winzerling, J.J. (2003) Iron-Binding Proteins in Insects. Department of Life Sciences. Presented at University of Wisconsin- Parkside, Parkside, WI.

Grants

Enhancing Obesity and Osteoporosis Research in Maine, USDA NRICGP

Can anthocyanin-rich foods prevent diabetes?, Gustavus and Louise Pfeiffer Research Foundation

Extruded Salmon Snack Utilizing Processing By-products, USDA NRICGP

Iron Metabolism of Mosquitoes, USPHS NIH GM56812. (2004)

Transcriptional Control of Ribonucleotide Reductase of Mosquitos, USPHS NIH (2003)