bluuebunny/crossref_metadata_2025 huggingface dataset:
The effects of Gpr1 and Gpa2 on the mating pathway in Saccharomyces cerevisiae
Authors: Duck Zachary A , Klouda Elisabeth , Haskins David , Johnson Janelle , Selcer Katie , Willhite Grant , Brigati Jennifer , Wright Stephen
Published: June 2021

Nutrient sensing and mating are clearly linked in a number of different fungal species. The mechanisms by which these two processes interact appear to differ in different species. We have observed that glucose sensing and mating are closely linked in Saccharomyces cerevisiae, with disruptions in the glucose sensing pathway causing reduction in mating pathway activity. In this study we sought to determine the mechanisms by which this reduction occurs. In a series of gene replacement experiments we have shown that overexpressing the mating receptor in glucose receptor (Gpr1) deficient cells restores the mating pathway. Likewise, overexpression of the g protein associated with glucose sensing, Gpa2, is able to restore the mating pathway in Gpr1蜐 cells. These effects seem to be independent of the activity of the g‐protein. Dominant negative effects were seen with some modified forms of Gpr1. We propose a mechanism of interaction that involves the formation of a complex involving potentially Ste2, Gpr1, Gpa2, and Gpa1. This research was funded by the NSF, Project #0952519
The silencing of GAPDH—an investigation of siRNA in an undergraduate biochemistry lab
Authors: Gabriel Scott E. , Wright Catherine A. , Backstrand Kyle M. , Temple Glena G.
Published: June 2021

This project outlines an 8‐week undergraduate laboratory which investigates the use of siRNA technology to silence a well‐known housekeeping gene, Glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) in a mammalian HeLa cell line. siRNA was discovered just over ten years ago, however there are few published reports on the use of this important technology in the undergraduate curriculum. This laboratory for upper level science majors, aimed to improve students’ mastery of modern day experimental techniques and strengthen their molecular biology lab skills by working with RNA, an inherently unstable molecule. Presented student data will showcase their successful efforts to knockdown GAPDH using siRNA through fluorescence, Western Blot and GAPDH assays. In addition to the experimental results, our course embedded assessment will show that students gained significant confidence in understanding and applying siRNA technology. This project was supported by a NSF CCLI grant #068183789.
Education of algae biofuels and phylogenetic trees for undergraduate students in a laboratory setting
Authors: Larson Sabreena , Jensen Travis , Keller Bryant , Patel Niraj , Bailey Cheryl
Published: June 2021

As alternative fuel research continues it is important to educate students as to be an active part of the scientific community. By integrating a new and interesting research area, such as algae biofuels, with a basic concept it is possible to have more student involvement and interaction. Here we use interactive worksheets combining basic knowledge of algae, DNA, and our basic concept: phylogenetic trees. The worksheets are supported by laboratory experiments where the students extract algal DNA, amplify a specific region, obtain the sequence and then build a phylogenetic tree with their obtained sequence and other sequences provided to identify the algal species. The laboratory module is being implemented at three different universities in Nebraska: Creighton University, University of Nebraska‐Kearney, and University of Nebraska‐Lincoln. The laboratory module is set up to cover multiple skills and education requirements so that it may fit in to a multitude of classes. Currently it is being implemented in a microbiology lab course, biochemistry lab course, and cellular biology lab course. This research is funded by: NSF EPSCoR‐ 1004–094
Formative study on the acceptance of Wheat Soy Blend Plus Plus (WSB++) as a daily complementary food supplement (CFS) in rural Bangladesh
Authors: Kramer Rebecca , Shamim Abu Ahmed , Kumkum Mehnaz Alam , Hanif Mohammad , Shaikh Saijuddin , Merrill Rebecca Day , Christian Parul
Published: June 2021

During a preparatory phase of a CFS feeding trial in rural NW Bangladesh, we field tested WSB++, an enhanced fortified blended food, using formative research methods to examine its acceptance by mothers and children and potential barriers to its use. Mothers (n = 44) living in the (JiVitA) study area with a child aged 6 to 18 months received pre‐cooked or uncooked WSB++ for 10 to 12 days. Acceptability and amount of WSB++ consumed were assessed through observation, interview and focus group discussion. Mothers liked feeding the WSB++ and found it as easy or easier to prepare as traditional complementary foods, but many added sugar or other spices to “improve the flavor”. Compliance (child consumed >;75% of the daily portion) was highest (100%) among mothers who cooked WSB++ at home with daily field worker visits, compared to those who prepared WSB++ without worker visits (63%) or who received pre‐cooked WSB++ (55%). Child illness was the main reason given for lower intake. Mothers reported reduced consumption of other complementary foods during WSB++ distribution compared to before, especially when the CFS was offered at usual mealtimes. Although diet replacement is a potential challenge for effective CFS use, mothers’ willingness to cook and feed WSB++, especially with some encouragement, suggests it is an acceptable CFS to use in rural Bangladesh.

Grant Funding Source: USDA (FANEP, NIFA), USA; WFP, Italy; DSM, Switzerland
The development of a reliable and feasible measure of school children's fruit and vegetable consumption
Authors: Taylor Jennifer C , Yon Bethany A , Johnson Rachel K
Published: June 2021

Feasible and reliable measurements of fruit and vegetable (F/V) consumption are needed that are based on children's actual consumption and are not subject to errors of children's self‐report. Our objective was to test the feasibility and reliability of three methods of assessing children's F/V consumption during lunch in two elementary school cafeterias. F/V consumption was assessed by weighed plate waste (WPW), direct observation (DO), and digital imaging (DI) during a training simulation and over ten school days. Feasibility was tested by the ability to assess F/V consumption for a target sample of at least 30–40 lunch trays during each school day. Interobserver reliability (IOR) was tested for DO and DI based on percent agreement for individual F/V item consumption and intraclass correlations (ICC) for total portions consumed per tray. WPW, DO, and DI assessed 340, 219, and 277 lunch trays respectively. Thirty‐one to 68 lunch trays were assessed during each school day, but no F/V items were selected on 15.2% of all trays. The net trays assessed for F/V consumption met or exceeded the target samples on all except one day, where 22% of participants did not select a F/V item. Percent agreement for DO was 93% and the ICC was 0.51. DI percent agreement was 96% and the ICC was 0.92. All methods were feasible for assessing F/V consumption during school lunch, but DI was more reliable than DO.

Grant Funding Source: USDA ‐ Vermont Agricultural Experiment Station and the Bickford Scholar Research Fund
Dietary Protein and Risk of Obesity and Central Adiposity in Middle‐aged and Older Adults in Framingham
Authors: Hasnain Syed Ridda , Buendia Justin Rene , Singer Martha R , Moore Lynn L
Published: June 2021

Aging triggers increases in fat mass and changes in body composition. Dietary protein may impact the balance of lean/fat mass, and the current protein RDA may be inadequate for older adults. Our goal was to examine the role of dietary protein on obesity and central adiposity in middle‐aged and older adults, using data from the Framingham Offspring Study, a prospective study of 5124 adults.

Dietary data were collected by 3‐day records for 1490 non‐obese (BMI<30kg/m2) healthy subjects. BMI and waist (umbilicus) were measured at each exam. ANCOVA and multiple logistic regression were used to control confounding by age, sex, education, height, activity, smoking, energy, % energy from fat and baseline weight.

Consuming more protein, alone or as part of a healthy diet (FV, fiber, whole grains) led to lower mean BMI and lower risk of obesity and central adiposity. Animal and plant protein intakes reduced obesity risk by 55% (OR: 0.45; 95% CI: 0.31–0.65) and 77% (95% CI: 0.15–0.34), respectively. Mean waist size declined (M: 40.9, 38.4, 36.2 cm; W: 34.7, 31.6, 29.4 cm; p<0.0001) with increasing protein intake. Higher total, animal and plant protein led to 71% (95% CI: 0.23–0.36), 49% (95% CI: 0.43–0.62) and 70% (95% CI: 0.24–0.37) lower central obesity risk, respectively. Effects were partly attenuated by adding baseline weight to the model. These results suggest that higher intakes of dietary protein may lower obesity risk in adults.

Grant Funding Source: Framingham Study funded by NHLBI; small analysis grant from ENC/USDA
Associations of magnesium intake with coronary artery calcification in the Framingham Heart Study
Authors: Hruby Adela , O'Donnell Christopher J. , Jacques Paul F. , Meigs James B. , Wood Richard J. , McKeown Nicola M.
Published: June 2021

Magnesium (Mg) may prevent vascular calcification, including calcification within atherosclerotic plaques underlying cardiovascular disease (CVD). We examined cross‐sectional associations of Mg intake, ascertained by food frequency questionnaire, on coronary artery (CAC) and abdominal aortic calcification (AAC) in men (N=1386, mean age 51±11 y) and women (N=1359, 55±11 y) free of CVD who participated in the Framingham Heart Study Multi‐Detector Computed Tomography (MDCT) Sub‐study. The Agatston score, a measure of calcium from CT scans, was ln‐transformed in sex‐specific Tobit models adjusted for covariates including age, BMI, major CVD risk factors and treatment for CVD risk factors, as well as intakes of calcium, vitamins D and K, saturated fat, fiber, alcohol, and energy, and hormone replacement therapy use in women. In women, a 50‐mg/d increment in Mg intake was associated with 24% lower CAC and 19% lower AAC (beta [SE]: −0.28 [0.11], P<0.01; −0.21 [0.10], P=0.05, respectively). Mg was not significantly associated with either CAC or AAC in men (−0.10 [0.08], P=0.18; −0.05 [0.08], P=0.51, respectively). The origins of the discrepant associations between men and women are unknown and require further study.

Grant Funding Source: American Heart Association Predoctoral Fellowship, the National Heart, Lung and Blood Institute's Framingham Heart Study (Contract No.N01‐HC‐25195), and the United States Department of Agriculture (USDA Agreement No. 58–1950‐7–707).
Why do WIC mothers choose not to follow current infant feeding recommendations?
Authors: Neshteruk Cody , Savage Jennifer , Birch Leann
Published: June 2021

Little is known about the perceptions and feeding practices of mothers choosing to formula feed. We examined the views and feeding behaviors of low‐income formula feeding mothers and obtained feedback about perceptions for a text message based feeding intervention. Focus groups (n=11) and phone interviews (n=20) were conducted with 68 women participating in the Pennsylvania Special Supplemental Nutrition Program for Women, Infants, and Children (WIC). Mothers in this study often exhibited inappropriate feeding behaviors such as feeding as a first response to infant distress and disregard for portion size recommendations. Mothers in this study indicated that they received conflicting advice from medical professionals, WIC, and family members. Many reported relying on their “instincts” and the notion that “all babies are different” when deciding whether or not to follow feeding guidance. When asked about what they would like with respect to delivery of an intervention, the majority of mothers were in favor of receiving information on a smart phone using text messages and websites. Interventions targeting this population should explore the use of smart phones and online media as modes of delivery. Curriculum should incorporate individualized messaging, tailor to social norms, and focus on continuity of information. Supported by PA TRACKS and CYFC at PSU.
Assessment of Nutrition and Physical Activity in Children Participating in the Shaping Healthy Choices Program
Authors: Scherr Rachel E , Linnell J , Nicholson Y , Spezzano T , Bergman J , Brian K , Briggs M , Cherr S E , Ermakov I , Feenstra G , Gellermann W , Hillhouse C , Nguyen L , Ontai L , Schaefer S , Smith M , Sutter C , Wright J , Zidenberg‐Cherr S
Published: June 2021

The Shaping Healthy Choices Program (SHCP) involves the integration of five components: nutrition education; family and community partnerships; regional agriculture; foods available on site; and school wellness policies. This study uses a pre/post‐test design to test the hypothesis that schools utilizing the SHCP, which includes garden‐based learning, will improve nutrition knowledge, science processing skills, and health related behaviors. Schools meeting inclusion criteria were randomly assigned to control or intervention groups. Dietary behaviors were assessed using digital photography plate waste measures, Block Food Frequency Questionnaires, and dermal carotenoid Resonance Raman Spectroscopy (RRS). Physical activity was measured using accelerometers. Nutrition knowledge and science processing skills were measured using validated questionnaires. Accuracy of plate waste was measured by 3 independent recorders; no statistical differences were observed. There was correlation between reported intake of MyPlate vegetable (MPV) servings and RRS (R2 = .31; p < .04). A slight correlation was observed between MPV and vegetable preferences at the intervention site (R2 = .03; p <.03). The assessment methods utilized in the SHCP and their interrelationships provide novel insight into conducting research in school‐based settings. Supported by UCANR and USDA 2011–38420‐20082.
Human Mammary Epithelial Cell Gene Expression and Product Secretion of Milk Lipids during Secretory Activation
Authors: Mohammad Mahmoud Ali , Haymond Morey W.
Published: June 2021

Background

The gene expression of lipid biosynthetic enzymes during initiation of lactation in humans in unknown.

Objective

To study mRNA expression of lipid metabolic enzymes in human mammary epithelial cell (MEC) in conjunction with the measurement of milk fatty acid (FA) composition during secretory activation.

Subjects

Gene expression from mRNA isolated from milk fat globule (MFG), a reflection of gene expression in MEC, and milk FA composition were measured from 6h to 42d postpartum in 7 normal women.

Results

Over the first 96h postpartum, gene expression of all aspects of lipid metabolism and milk FA production increased several folds, including: lipolysis at the MEC membrane, FA uptake from blood, intracellular FA transport, de novo FA synthesis, FA and glycerol activation, FA elongation, FA desaturation, synthesis of triglycerides, cholesterol synthesis and lipid droplet formation. Gene expression of the key lipid synthesis regulator, sterol regulatory element‐binding transcription factor1 (SREBF1), increased 2.0‐fold by 36h and remained elevated over the study duration. The mRNA expression of thyroid hormone responsive (THRSP) and insulin‐induced2 (INSIG2) genes increased progressively to plateau by 96h. In contrast, mRNA expression of peroxisome proliferator‐activated receptor gamma (PPARG) decreased several folds. With the onset of lactation, de novo synthesis of FAs (C6‐C12) was the most prominent change in milk FA composition and mirrored the expression of FA synthesis genes.

Conclusion

Milk fat synthesis and secretion in humans is a complex process requiring the orchestration of a wide variety of pathways of which SREBF1 may play a primary role.

Grant Funding Source: USDA/ARS Grant # 6250–51000
High protein diets enhance body composition in rats: a comparative analysis of milk‐and soy‐based energy restricted diets
Authors: Pasiakos Stefan M , Wulff Charles R , Murphy Nancy E , Gaffney‐Stomberg Erin , Young Andrew J , McClung James P
Published: June 2021

Consuming high protein diets during energy restriction may protect muscle health. However, the extent to which protein source modulates muscle responses to energy restriction is not well described. Eighty male Sprague‐Dawley rats (12 weeks old) were randomly divided into 8 treatment groups and fed energy adequate (EA) and energy restricted (ER; 60% of the ad libitum feed intake for EA) diets containing two levels (control protein: CP, 10%; and high protein: HP, 35%) and sources (milk protein concentrate: MPC; and isoflavone‐free soy isolate: SOY) of protein for 16 weeks. Body mass was assessed weekly, and fat (FAT) and fat‐free mass (FFM), as percentages of total body mass were determined on weeks 0 and 16 using dual energy x‐ray absorptiometry. Body mass was similar for all rats on week 0, and increased (P < 0.05) linearly independent of protein level and source. Subsequent to week 0, body mass was lower for ER compared to EA (energy‐by‐time, P < 0.05). Protein level influenced FAT and FFM independent of protein source and energy, as FAT was lower and FFM was higher at week 16 for HP compared to CP (P < 0.05). Regardless of protein level and source, FAT was lower and FFM was higher at week 16 for ER than EA (P < 0.05). These data suggest consuming a HP diet may spare FFM and improve body composition independent of energy status, although skeletal muscle responses to milk‐and soy‐based diets are likely comparable. Supported by Dairy Research Institute and USAMRMC
Identification of bean polyphenols that inhibit and enhance iron uptake by Caco‐2 cells
Authors: Hart Jonathan J. , Glahn Raymond
Published: June 2021

Polyphenolic compounds present in bean seed coats are known to be inhibitors of Fe bioavailability; however, the specific polyphenols in bean seed coats that inhibit Fe bioavailability have not been identified. In addition, the antioxidative properties of some polyphenols may actually promote Fe absorption. The objective of this study was to identify the major polyphenols that affect Fe bioavailability in black beans. Extracts of white beans and black beans were evaluated for effect on Fe uptake using a Caco‐2 cell model. LC/MS was then used to identify compounds present in active extracts. As expected, Iron uptake was inhibited by extracts from black bean seed coats relative to that of white beans. Examination of the effect of individual polyphenols present in black bean seed coats showed varying dose related Fe uptake responses, ranging from inhibition to stimulation. Myricetin, quercetin and quercetin 3‐glucoside inhibited iron uptake; ferulic acid and salicylic acid had no effect; and caffeic acid, epicatechin, gallic acid and kaempferol stimulated iron uptake in Caco‐2 cells. The results clearly indicate that not all polyphenols inhibit iron uptake and that some are promoters. Targeting these compounds in breeding and or processing of beans could yield improvements of Fe bioavailability. Funded by USDA‐ARS.
Differential effects of hepcidin on heme and nonheme iron absorption in a rat model of iron overload
Authors: Cao Chang , Thomas Carrie , Insogna Karl , O'Brien Kimberly
Published: June 2021

Heme and non‐heme iron (Fe) are taken up by enterocytes via distinct transporters but may share the same exporter protein, ferroportin (FPN). The Fe regulatory hormone, hepcidin, controls non‐heme Fe absorption by inducing FPN degradation. Whether hepcidin affects heme Fe absorption is largely unknown. To address this question, we utilized stable Fe isotopes to measure heme and non‐heme Fe absorption in a rodent model of Fe overload. Male Sprague Dawley rats received bi‐weekly injections of PBS (control), 4 mg Fe (moderate Fe), or 12 mg Fe (high Fe) to induce Fe overload. After 2 weeks of treatment, rats were gavaged on alternate days with 57Fe as labeled ferrous sulfate and 58Fe as labeled porcine RBCs. Red cell incorporation of 57Fe and 58Fe was assessed 10‐days post‐dosing. Fe overload in rats increased Fe concentrations in liver (p=0.008) and spleen (p=0.002), increased levels of liver hepcidin transcripts (p=0.005), and reduced absorption of both heme (p=0.004) and nonheme Fe (p=0.01) by 69% and 63%, respectively. While humans absorb heme Fe more efficiently, non‐heme Fe absorption was 2.4‐times higher than heme Fe absorption in the rat (p=0.0009). Liver hepcidin had a greater association with non‐heme compared to heme Fe absorption (p=0.02, ANCOVA). Hepcidin up‐regulation in the rat inhibits both non‐heme and heme Fe absorption but suppresses heme Fe absorption less effectively.

Grant Funding Source: USDA 2008–0857

Grant Funding Source: USDA
Vitamin D and calcium supplementation decrease body and fat weight gain and improve biomarkers of adiposity in diet‐induced obesity
Authors: Song Qingming , Sergeev Igor N
Published: June 2021

Low vitamin D status (circulating concentration of 25(OH)D) and low calcium intake have been linked with an increased risk of obesity, however the underlying molecular mechanisms of vitamin D and calcium in obesity are not understood. To investigate effects of calcium and vitamin D supplementation on the development of obesity, male C57BL/6J mice (4 weeks of age) were fed for 10 weeks either a high fat (HF) diet (60% energy from fat) or HF diets containing 1.2% calcium, 10000 IU/kg vitamin D3 and their combination. The results obtained demonstrated that the calcium plus vitamin D3 supplementation reduced the body and adipose tissue (epididymal and retroperitoneal fat depots) weight gain and decreased the plasma glucose and insulin concentrations, as compared with the HF diet group. In addition, a high vitamin D3 intake significantly raised the plasma concentrations of 25(OH)D and its hormonal form, 1,25(OH)2D, in HF diet‐fed mice. These findings indicate that increased dietary vitamin D and calcium intakes improve biomarkers of adiposity, insulin sensitivity, and vitamin D status in diet‐induced obesity and suggest a role for vitamin D and calcium in preventing body and fat weight gain. Supported by USDA 2009–35200‐05008 and USDA SD00H325 to INS.
Dysregulation of Vitamin K Homeostasis in Brain, not Heart of Vitamin E‐Injected rats Fed Deuterium‐labeled Phylloquinone
Authors: Farley Sherry Mae , Leonard Scott W , Kirkwood Jay S , Stevens J Frederick , Traber Maret G
Published: June 2021

Subcutaneous (SC) α‐tocopherol (α‐T) injections in rats resulted in significant reductions in phylloquinone (PK) and menaquinone‐4 (MK‐4) in many extra‐hepatic tissues. To investigate the mechanism for vitamin K loss, rats were fed deuterium‐labeled (d4)‐PK for 17 days; starting on day 10, they were SC injected for 7 days with saline, vehicle, or α‐T (100 mg/kg BW), then on day 8, plasma and tissues were collected. After α‐T injections, liver α‐T was increased 10‐fold and its carboxychromanol catabolite (CEHC) increased 40‐fold, while brain α‐T was unchanged, brain CEHC increased from undetectable to 300 ± 60 pmol/g. Brain vitamin K was ~70% labeled; it was enriched in d4‐MK‐4 (14 pmol/g) relative to d4‐PK (1.5 pmol/g); α‐T injections decreased d4‐PK to 67% and d4‐MK‐4 to 45% compared with controls; PK conversion to MK‐4 was similar between groups. In contrast, heart contained ~85% enrichment in d4‐PK, while α‐T injections had no significant effects on heart total vitamin K. The high heart d4‐PK concentrations (~40 pmol/g) may protect it from the deleterious α‐ T effects. In brain, injections decreased both d4‐MK‐4 and d4‐PK concentrations suggesting that either uptake of d4‐PK or recycling of d4‐MK‐4 was potentially inhibited by CEHC.

Grant Funding Source: National Research Initiative Grant 2009–35200‐05031 from the USDA
Identification of Anthocyanins in Purple‐fleshed Sweetpotato and Stability during Various Cooking Conditions
Authors: Xu Jianteng , Su Xiaoyu , Lim Soyoung , Griffin Jason , Carey Edward , Katz Benjamin , Tomich John , Wang Weiqun
Published: June 2021

We previously selected a purple‐fleshed sweetpotato P40 that prevented colorectal cancer in a murine model. This study is to identify anthocyanins and tested stability. P40 contains anthocyanins up to 13 mg/g dry matter. Total 12 acylated anthocyanins have been identified by HPLC‐MS/MS. The top three anthocyanins are cyanidin 3‐caffeoyl‐p‐hydroxybenzoyl sophoroside‐5‐glucoside, peonidin 3‐caffeoyl sophoroside‐5‐ glucoside, and cyanidin 3‐(6″ ‐caffeoyl‐6″‐feruloylsophoroside)‐5‐ glucoside. Seven acylated cyanidin species and five peonidin species contribute for 69% and 31% of total anthocyanins, respectively. Over 80% belong to cyanidin as measured by acid hydrolysis. Therefore, as a cyanidin‐predominated variety, P40 is unique when compared with other purple‐fleshed sweetpotatoes that usually contain more peonidin than cyanidin. While baking does not impact overall anthocyanins, steaming, high pressure cooking, microwaving, and frying significantly reduce 20% of total anthocyanins. Mono‐acylated anthocyanins show a higher resistance against heat than di‐ and non‐acylated. Among of which, cyaniding 3‐p‐hydroxybenzoylsophoroside‐5‐glucoside exhibits the best thermal stability. Better understanding of sweetpotato anthocyanins and stability may lead the development of a functional product for health benefits (supported by USDA Cooperative KS410–0214022).
Menaquinones content of human serum and feces
Authors: Karl J Philip , Fu Xueyan , Saltzman Edward , Meydani Simin N , Meydani Mohsen , Barnett Junaidah B , Booth Sarah L
Published: June 2021

Bacterially‐synthesized menaquinones (MKn) may contribute to vitamin K (VK) nutriture. There are limited data on interindividual variability in endogenous MK synthesis and its relation to circulating forms of VK. Serum and fecal VK concentrations were assessed in 13 healthy adults (45–65 yr) consuming a standardized diet for 14 d. Phylloquinone (PK), MK4 and MK6‐MK13 concentrations were measured by HPLC in fasting serum collected on day 8 and 72 hr‐fecal homogenates collected days 8–10. LC/MS was used to confirm individual MKn. PK, the primary dietary form, comprised only 6 ± 3% of total fecal VK content (fecal PK [mean ± SD]; 0.45 ± 0.17 μg/g dry wt), and was not correlated with serum PK concentrations (1.85 ± 0.89 pmol/L). No MKn were detected in serum. MK4 and MK6‐MK12 were detected in all, and MK13 in 7 fecal homogenates. Mean fecal MKn concentrations ranged from 0.17 ± 0.05 μg/g dry wt for MK4 to 4.95 ± 2.30 μg/g dry wt for MK10, with MK10 comprising 63 ± 19% of total fecal MKn content. Based on fecal contents, we conclude that the majority of MKn isoprenologues are present in the human colon, but with evidence of interindividual variation despite 10 d of diet standardization. The clinical implications of these findings, and the role of MKn in supporting VK nutriture are yet to be determined. Funded by a grant from General Mills and USDA contract #58–1950‐7–707.

Grant Funding Source: USDA
Characterization and bioaccessibility of tangeretin‐loaded zein colloidal system
Authors: Chen Jingjing , Zheng Jinkai , McClements David Julian , Xiao Hang
Published: June 2021

Tangeretin, a citrus flavonoid has been shown to possess many beneficial bioactivities. However, bioaccessibility of tangeretin is relatively low due to its high hydrophobicity, which limits its absorption in gastrointestinal tract. The aim of this study was to design a delivery system to improve bioaccessibility of tangeretin. Tangeretin‐loaded zein colloidal system with an average particle size of 400 nm was produced by mixing aqueous phase containing β‐lactoglobulin and organic phase containing zein and tangeretin. After removal of organic solvent, particle size was decreased to about 300 nm. The influence of temperature, pH, ionic strength on the stability of the system was examined. Apparent particle aggregation occurred at high temperature (70 °C), low ionic strength and around isoelectic point of β‐lactoglobulin. Powder form of the above delivery system was successfully produced by lyophilization. After reconstitution into an aqueous solution, the delivery system maintained similar responses to the environment stress as freshly prepared one, suggesting an excellent applicability of the system as a convenient powder ingredient. Using a simulated gastrointestinal model, we demonstrated the colloidal system significantly enhanced tangeretin bioaccessibility. In conclusion, we successfully developed a stable colloidal system that can potentially improve bioaccessibility of tangeretin.

Grant Funding Source: EPA‐USDA‐NSF grant
Blueberries and strawberries activate neuronal housekeeping in critical brain regions of stress‐induced young rats.
Authors: Poulose Shibu , Bielinski Donna F , Carrihill‐Knoll Kirsty , Rabin Bernard , Shukitt‐Hale Barbara
Published: June 2021

Dysfunctional autophagy, where accumulation of damaged or complex cellular components in neurons in response to sublethal cell stress, has been implicated in an array of brain disorders. This phenomenon plays a pivotal role in aging because of the increased vulnerability of the aging brain to increased oxidative stress and inflammation. Blueberries and strawberries are rich in neuroactive compounds, and diets supplemented with these berries have been shown to improve behavior and cognitive functions in young rats which were subjected to oxidative and inflammatory stress by exposure to irradiation with high energy and charge (HZE) particles, a model for accelerated aging. The current study explored the neurochemical changes in the brain, particularly autophagy, a neuronal housekeeping mechanism. Irradiation disrupted protein homeostasis as evidenced by increased accumulation of ubiquitinated proteins in hippocampus and striatum. Feeding animals with either berry diet (2% w/w), prior to irradiation as well as 30 days post irradiation, protected these brain regions against inflammation and oxidative stress. The berry diets also enhanced the clearance of ubiquitinated proteins measured in terms of the levels of p62, beclin1, ATG7, and MAP1B‐LC3, and phosphorylation of mTOR, indicating activation of autophagy. This study extends molecular evidence for the health‐promoting properties of berries.

Grant Funding Source: USDA intramural funds and NASA grant NNX13AB73G
Tomato powder, not tomato extract or lycopene, attenuates alcohol‐induced hepatic inflammatory foci and steatosis in rats
Authors: Stice Camilla Peach , Liu Chun , Greenberg Andrew S , Ausman Lynne M , Wang Xiang‐Dong
Published: June 2021

Alcoholic fatty liver disease (AFLD) is a major cause of morbidity and mortality worldwide. Consumption of tomatoes, rich in the carotenoid lycopene (LYC), is associated with decreased risk of chronic disease. However, our previous results show a negative interaction between alcohol consumption and supplementation with LYC. The present study aims to investigate the potential protective effects of tomato powder (TP, representative of whole tomato) and tomato extract (TE, lipid soluble tomato components) against AFLD development in male Wistar rats. Animals were group pair‐fed with Lieber‐DeCarli liquid diet (LD) or alcoholic liquid diet (ALD, 36% energy from ethanol) and supplemented with LYC, TP, or TE to provide 1.1 mg/kg BW LYC for 4 weeks. We observed a significant increase in both hepatic steatosis and inflammatory foci in rats consuming ALD as compared to LD. LYC supplementation had no effect on the degree of steatosis and, consistent with our previous results, significantly increased inflammatory foci in rats fed ALD. Interestingly, dietary TP significantly reduced the severity of both steatosis and inflammatory foci in rats fed ALD, whereas TE had no effect on either outcome. This study reveals the protective effects of TP supplementation against AFLD development and further highlights the potential detrimental interaction of alcohol and supplementation with a single nutrient in replace of whole food.

Grant Funding Source: USDA 1950–51000‐064S
Individual genetic variations (IGV) of β,β‐carotene monooxygenase‐1 (BCMO1) are associated with changes in total cholesterol in young Mexican adults
Authors: Marques Courtney , Teran‐Garcia Margarita , Erdman John W. , Vargas‐Morales Juan Manuel , Aradillas‐Garcia Celia , Group UP‐AMIGOS
Published: June 2021

Retinoids are key in facilitating many physiological functions, including lipid metabolism, and are typically formed from the cleavage of β,β‐carotene by BCMO1. Recently, an IGV in the BCMO1 gene (SNP, rs10048138) was associated with cholesterol levels in a U. S. cohort; however, there are no replicates in the Mexican population. Our objective was to examine the association of this BCMO1 marker with cholesterol levels, a risk factor for metabolic disease, in college‐age individuals from the UP‐AMIGOS cohort. UP‐AMIGOS is a multidisciplinary project on genetics, obesity, and social environment between the Universities of San Luis Potosi (San Luis Potosi, Mexico) and Illinois. For this cross‐sectional study genotype, health and nutrition data from 72 participants (aged 18–21 years) was analyzed. Homozygotes for the minor allele of BCMO1‐rs10048138 had lower cholesterol (137.4±9.8 mg/dL) compared to the other genotypes (158.1±2.7 and 157.6±4.0 mg/dL) adjusted for sex and age (p<0.04). Our analyses show that genetic variability in BCMO1 was associated with cholesterol levels and may prove to influence risk for metabolic disease; this association was strengthened (p<0.02) when considering smoking.

Grant Funding Source: USDA/NIFA Hatch Project (#600108–793000‐793323 and #600109–698000‐698354)
Synergistic inhibition of lung cancer cell proliferation by 4′‐ hydroxylnobiletin and atorvastatin
Authors: Charoensinphon Noppawat , Zheng Jinkai , Dong Ping , Qiu Peiju , Xiao Hang
Published: June 2021

Combination treatment with different anticancer agents may produce enhanced effects due to their synergistic interaction. Previously, we reported that nobiletin (NBT, a flavonoid found in citrus fruits) and atorvastatin (ATST, Lipitor, a lipid‐lowering drug) in combination produced strong synergy in inhibiting lung cancer cell growth. Herein, we further investigated the interaction between ATST and a major metabolite of nobiletin, 4′‐ hydroxylnobiletin (4HN) in lung cancer cells. Our results showed that 4HN produced much stronger growth inhibition on cancer cells than its parent compound NBT. Moreover, 4HN/ATST co‐treatments at low doses consistently produced much stronger inhibitory effects on different cancer cells in comparison to those produced by 4HN or ATST alone at higher doses. Isobologram analysis confirmed that the interaction between 4HN and ATST was highly synergistic. Our results further demonstrated that the synergistic growth inhibition on lung cancer cells by 4HN/ATST co‐treatment was associated with extensive G0/G1 cell cycle arrest and apoptosis, as the results of modulating key oncogenic signaling proteins by 4HN/ATST co‐treatment. Taken together, our results showed a strong synergy between ATST and 4HN in inhibiting lung cancer cell growth, which provided a strong rationale to use NBT and ATST in combination for lung cancer inhibition.

Grant Funding Source: AICR, NIH, and USDA
Apigenin and naringenin decrease cell number in a dose dependent manner in non‐transformed young adult mouse colonocytes (YAMC) but not in those expressing a dominant negative p53‐mutant (mp53 YAMC)
Authors: Daniels Wesley Danielle , Allred Kimberly F. , Allred Clinton D. , Patil Bhimanagouda S. , Turner Nancy D.
Published: June 2021

We have shown that apigenin and naringenin (flavonoids) suppress colon carcinogenesis by inducing apoptosis and suppressing proliferation in rats. The aim of this study was to determine if the mechanism of action was p53 dependent. YAMC and mp53 YAMC cells were treated with apigenin (0.1 μM, 1μM, 10 μM), naringenin (0.1 μM, 1 μM, 10 μM, 25 μM, 50 μM), or estradiol (1 nM, positive control) in DMSO for 4 d at non‐permissive conditions. Cells were counted and proliferation measured using Brd‐U. YAMC cells treated with 1 μM or greater apigenin or naringenin exhibited dose‐dependent decreases (P<0.005) in cell numbers compared to DMSO. No differences in cell number were identified in the mp53 YAMC cells except with the highest concentration of apigenin and naringenin (P < 0.0001). No differences in proliferation were observed with apigenin or naringenin in either cell line. The dose‐dependent reduction in non‐transformed cell number induced by apigenin and naringenin are in part p53‐mediated. The reduction of cell numbers in mp53 YAMC cells resulting from the greatest concentrations suggests alternate pathways can be induced by these flavonoids. However, the effect on cell number is not linked directly to suppression of colonocyte proliferation in either p53 competent or mutant colonocytes. Funded by USDA/VFIC 2010–34402‐20875.
Identification of variants in the fatty acid desaturase (FADS) gene cluster associated with total cholesterol and HDL‐C in young Mexican college students. UP‐AMIGOS study.
Authors: Vazquez‐Vidal Itzel , Teran‐Garcia Margarita , Andrade Flavia C.D. , Aradillas‐Garcia Celia , Vargas‐Morales Juan Manuel
Published: June 2021

A recent genome‐wide association study (GWA) in the Mexican population has identified several genetic loci associated with blood lipid levels in adults. The FADS gene cluster has not been studied in youngsters. Our aim was to report the single‐nucleotide polymorphism (SNP) in the FADS gene cluster associated with high TC, and low HDL‐C in young Mexicans from the college‐age UP‐AMIGOS cohort. Anthropometrics, smoking habits, cardiovascular disease history, and serum lipid profile were determined (n=72, aged 18–21 years). Genotyping was conducted using the Illumina system. Genotype‐phenotype associations were assessed by multiple linear regression models adjusted by sex and age. Genetic variability in the FADS gene was associated with TC (6 SNPs) and HDL‐C (2 SNPs) levels. Our data provides new valuable information for future studies where other environmental factors (diet, exercise) are taken into consideration to better understand the onset of dyslipidemia. It also provides useful data for early identification of individuals at risk and for comparative studies in different population.

Grant Funding Source: USDA/NIFA Hatch Project (#600108–793000‐793323 and #600109–698000‐698354)
Accelerated growth during infancy alters bone characteristics and the activity of mesenchymal stem cells (MSC) in neonatal piglets
Authors: Seabolt Brynn , Li Yihang , Stahl Chad
Published: June 2021

Rapid growth during infancy has been associated with increased risk of childhood obesity. We examined the effects of increased growth rate caused by overfeeding on growth, bone integrity, and activity of MSC in neonatal piglets. Limit fed (LF) animals received a standard piglet milk replacer at a rate to achieve normal growth, while the overfed (OF) group voluntarily consumed nearly 60% more of the same diet for 21 days. Overfed piglets had higher growth rate (P < 0.001) and feed intake (P < 0.05), and lower feed efficiency (P < 0.001) compared to LF piglets. The OF animals had heavier, larger bones with greater bone mineral content than the LF group (P < 0.05). Surprisingly, MSC isolated from OF animals adopted an adipocytic lineage less readily than those isolated from LF animals after induction with adipocytic media. This was indicated by lower gene expression of PPARG, LPL and AP2 (P < 0.05) as well as by decreased lipid accumulation determined by Oil Red O staining. Increased (P < 0.2) gene expression of Runx2 following osteogenic induction was also seen in MSC from OF pigs. Accelerated growth during infancy led to a decreased response of MSC to an adipocytic environment, as well as an increased response to osteogenic induction. These results indicate the potential for neonatal dietary interventions to influence growth and body composition later in life via programming of MSC.

Grant Funding Source: NC Agricultural Research Service
Phosphoinositide‐3‐kinase regulation of neutrophil mechanosensing is context dependent
Authors: O'Brien Xian , Oakley Katie E , Loosley Alex J , Tang Jay X , Reichner Jonathan S
Published: June 2021

To localize to an injury or infection, neutrophils interpret biochemical and physical cues. Previously, we have shown that neutrophil chemotaxis on fibronectin (Fn) is also regulated by mechanical stiffness through a phosphoinositide‐3‐kinase (PI3K) dependent mechanism. To determine whether substrate stiffness is a universal regulator of function, we quantified human neutrophil migration dynamics on fibrinogen (Fgn), collagen (Col), and Fn, integrin ligands encountered during an inflammatory response. As an inflammatory matrix includes multiple ligands, we also analyzed neutrophils on mixtures of Fn and Fgn. Analysis of mean square displacement, rate of migration, chemotactic index and cell polarity identified ligand‐specific mechanosensitivity influenced by both beta 1 and 2 integrins. Stiffness‐dependent changes in migration and cell polarity varied by ligand, with mechanospecific changes in migration observed on Fn, Col, and mixtures of Fn and Fgn, but not Fgn alone. Inhibitor studies show the role of PI3K in regulating neutrophil polarization and mechanotactic migration dynamics increases in complexity with the model system. Our data show that PI3K can act to either facilitate or inhibit neutrophil chemotaxis toward fMLP with protein substrate and stiffness determining this contextual regulation. Supported by grants from the NIH, the NSF's MRSEC at Brown, and RI Hospital.
Edge assay: kinetic analysis of reagents affecting cell clumping
Authors: Zem Gregory , Ter‐Papyan H , Arvizu S , Nernsuan J , Rabizadeh S , Amidi M , Nazari S , Waas S , Lee S , Orujyan A , Manookian L , Taghinia P , Carpio C , Trinh D , Balazadeh H , Oppenheimer S B
Published: June 2021

Cells in clumps are often more dangerous than single cells in a cancer setting and in infectivity. We present a fine‐tuned method to test reagents that may affect cell clumping. In each experiment two 0.2ml droplets of distilled water were placed on glass microscope slides to which 1% formaldehyde fixed yeast (Saccharomyces cerevisiae) in distilled water (pH adjusted to 4.0) were added at dilutions that resulted in 8–25 single yeast and clumped yeast cells that were observed at the edge of the droplets, providing a standardized reference point for counting single vs clumped (two or more) yeast. Using birchwood toothpicks, various concentrations of solid reagents (salts, sugars, complex carbohydrates, amino acids) (1–10 mg) were added to the experimental drops and nothing was added to the control drops. Numbers of single and clumped yeast were counted before addition of reagents, immediately after addition and at 20, 40, 60 min after reagent additions in 1179 trials (21 C). The numbers of single yeast and yeast clumps were observed with a light microscope (100–200 X) and recorded at each step. The percentages of single yeast in experimentals and controls were plotted showing SEM error bars and t‐tests were performed at 20,40,60 min comparing the percentage of single yeast in experimentals vs controls. P values of less that 0.05 suggested that the percentages of single cells in the experimentals were significantly different than in the controls. The results show that the precise analysis that this assay affords is able to detect very small yet statistically significant differences in reagent effects and could lead to identification of reagents that may be further developed into anti‐clumping drugs (Supported by NSF Presidential Award 0731633, NIH SCORE, RISE, MARC, the Joseph Drown Foundation, the Sidney Stern Memorial Trust).
Use of Platelet Proteomic Profiles to Distinguish Platelets Activated by Different Agonists
Authors: Yin Wei , Shanmugavelayudam Saravan Kumar , Rubenstein David Alan
Published: June 2021

The objective of this study was to identify platelet signature proteomic profiles that would distinguish platelets activated by mechanical agonists and biochemical agonists. Human platelet rich plasma were treated with TRAP or exposed to various shear stress waveforms at physiological and pathological level. Plasma proteins and platelet lysate were then applied to CM‐10 ProteinChip arrays and analyzed using a SELDI ProteinChip reader. In the low molecular range (2,000 to 10,000 M/Z), more than 100 protein/peptide peaks were identified, and platelets responded to TRAP and shear stress differently. For platelet proteins, TRAP not only modified the magnitudes of significantly differentiated protein peaks, but also altered the shape of the spectral patterns. Shear stress only affected the magnitudes of differentiated protein expression, but did not cause the spectral pattern change. For plasma proteins, TRAP treatment did not cause any changes. Shear stress led to a significant decrease in peak intensity at M/Z=7765, while the same peak increased significantly in platelet proteins, indicating platelets might have sequestered this protein in response to shear stress stimulation. Preliminary studies suggested that this protein was possibly platelet factor 4. This work is supported by NSF 1039913.
Euglycemia following IED‐like blast exposure
Authors: Scrimgeour Angus Gray , Joosen Marloes J. , Young Andrew J. , van Helden Herman H.
Published: June 2021

Episodes of hyperglycemia are considered to be a secondary insult in traumatic brain injury (TBI) patients and have been shown to be associated with impaired outcome. Intensive insulin therapy and strict monitoring of blood glucose (BG) during neurointensive care is considered crucial to maintain the brain's glucose demand after brain injury, and to avoid both hypo‐ and hyper‐glycemia. This study investigated glucose levels in peripheral blood in the acute phase following IED‐like blast injury.

METHODS

Twenty‐two anesthetized adult Wistar rats were exposed to a blast induced by an explosive charge (18g Semtex; 120kPa incident pressure; 12 sec positive phase duration) in a 40×40cm blast‐tube; 22 anesthetized, non‐exposed rats served as Controls. Tail blood was sampled at 24h, 48h and 14 days post‐blast. BG was determined using a commercial photometric analyzer.

RESULTS

All blast‐exposed rats exhibited transient bradycardia and bradypnea post‐injury, and loss‐of‐conciousness averaged 3min 20sec. Blast induced deficits in spatial learning and memory, and impaired vestibular motor activity. Post‐blast BG values ranged from 4.8 to 7.3 mmol/l over 14 days. No biochemical hypoglycemia (BG <3.3 mmol/l) was observed. Stress hyperglycemia (BG >;10.0 mmol/l) occurring during an acute illness in those with previously normal glucose tolerance was not observed.

CONCLUSIONS

Blast‐induced TBI did not induce hyperglycemia in this rat model, and hence there is no support for the use of intensive insulin therapy, in contrast to non‐blast TBI patients with hyperglycemia.

Funded by the U.S. Army MRMC and U.S. Air Force MSA.
Chronic stress alters brain and peripheral glucocorticoid and mineralocorticoid receptors in a wild bird
Authors: Lattin Christine Renee , Romero L. Michael
Published: June 2021

Although sustained high glucocorticoid titers can result in pathology, the effects of chronic stress on peripheral tissue sensitivity to glucocorticoids are not well understood. In this study, we subjected house sparrows (n=24) to a 3‐week standardized chronic stress protocol followed by a week of recovery and compared their glucocorticoid and mineralocorticoid receptor binding to that of control birds (n=18). We examined receptors in 9 tissues: whole brain, liver, muscle, fat, kidney, spleen, skin, testes and ovary. Animals were sacrificed weekly in the stress group, and every other week in the control group. Unlike previous work, which has typically demonstrated decreased tissue sensitivity in response to sustained high glucocorticoid titers, we found that chronic stress either did not affect receptor binding (skin, fat, liver and spleen) or increased receptor binding in a time‐and tissue‐dependent manner (muscle, brain, kidney, ovary and testes). Most chronic‐stress induced effects disappeared after a week of recovery, but receptor binding in pectoralis muscle remained high, suggesting ongoing metabolic disruption. Work supported by NSF, EPA, and Tufts University.