bluuebunny/crossref_metadata_2025_split huggingface dataset:
Assessing nutrition knowledge using Healthy Plate photo method among youth participating in WeCook
Authors: Chai Weiwen , Walther Ashley , Dunker Tara , Franzen‐Castle Lisa , Krehbiel Michelle 
Published: June 2021

ObjectiveMounting evidence suggests that poor dietary habits are linked to a lack of cooking at home and insufficient nutrition knowledge such as making a healthy plate. This study assessed the changes in nutrition knowledge among youth participating in WeCook.MethodsTwenty‐four 4th and 5th grade students from two Title I elementary schools in Nebraska were recruited to participate in WeCook, an on‐going, semester‐based, 12‐week after‐school prevention program focusing on youth cooking and physical activity. Surveys were administered at each site before and after the program. Youth's nutrition knowledge was assessed using a novel Healthy Plate photo method (based on MyPlate guidelines) and previously validated instrument. Youth were asked to build a healthy plate using a blank MyPlate template and food models pre‐ and post‐intervention, with healthy plate photos taken by the researchers. Healthy plate photos were assessed using a scoring system based on the five food groups present on healthy plate; one point was assigned to each of the correct food groups present with a maximum total score of five. The differences in pre‐ and post‐intervention scores of healthy plate photos and related nutrition knowledge were assessed using paired t‐test and chi‐square test.ResultsThe average score of correct food groups on healthy plate photos was significantly increased after WeCook (Pre: 3.8±0.8, Post: 4.4±0.8; P=0.003). Fifty‐six percent of participants received the maximum score (5 points) after the program whereas only 17% scored five pre‐ program. Additionally, there were non‐statistically significant increases in knowledge on daily fruit and vegetable consumption and the importance of breakfast after the program.ConclusionOur preliminary results suggest WeCook appears effective to improve nutrition knowledge among 4th and 5th grade participants in the study.Support or Funding InformationThis material is based upon the work of CYFAR University of Nebraska‐Lincoln Sustainable Community Project, a project supported by the National Institute of Food and Agriculture, United States (U.S.) Department of Agriculture, through a cooperative agreement with the University of Nebraska‐Lincoln under award number 2014‐41520‐22207.
Identification of Glycoproteins on the Surface of Bovine Milk Exosomes and Intestinal Cells that Facilitate Exosome Uptake in Human Colon Carcinoma Caco‐2 Cells
Authors: Sukreet Sonal , Zhang Hanyuan , Adamec Jiri , Cui Juan , Zempleni Janos 
Published: June 2021

BackgroundExosomes are small, cargo‐containing vesicles that are secreted by donor cells to elicit changes in gene expression and metabolism in recipient cells. In previous studies, we provided evidence that, in addition to endogenous synthesis, exosomes may also be obtained from dietary sources such as bovine milk, and intestinal transport is facilitated by endocytosis.HypothesisGlycoproteins on the surface of bovine milk exosomes and human intestinal cells play essential roles in exosome transport.MethodsExosomes were isolated from bovine milk by differential centrifugation and authenticated as recommended by International Society for Extracellular Vesicles (Fig. 1). Surface proteins or glycan modifications in exosomes or Caco‐2 cells were removed using proteases (Glu‐C, trypsin, Arg‐C, Asp‐N, or proteinase K) or glycosidases [N‐glycosidase F (PNGase F), galactosidase, O‐glycosidase, neuraminidase, N‐acetyl glucosaminidase, or a combination of all]. Controls were incubated with solvent. Exosomes were labeled with FM4–64, and unlabeled fluorophore was removed. Surface peptides released after the treatment were identified using LC/MS‐MS, Mascot and Sequest databases. Glycoproteins on the external surface were identified using a series of tools based on neuronal networks including NetNglyc, NetOglyc, NetCglyc 1.0 and GlycoEP. Transport kinetics were modelled using the Michaelis Menten equation. One‐way ANOVA and Bonferroni's multiple comparison were used for statistical analyses.ResultsWhen exosomes or cells were treated with individual proteases (Fig. 2) or glycosidases (Fig. 3), exosome uptake decreased up to 80% when compared to controls even at prolonged incubation times. Treatment with galactosidase, PNGase F and O‐glycosidase had a stronger effect than other glycosidases. When exosomes or cells were treated with the glycosidase mixture, no uptake was detected or uptake was decreased by 80%, respectively, in comparison to controls (Fig. 4). 149 total proteins were identified in bovine exosome, including 4 (N), 2 (O) and 2 (C) glycan binding sites; 298 proteins were identified in Caco‐2 cells, including 46 membrane proteins that included 29 (N), 33 (O) and 6 (C) glycan binding sites.ConclusionsGlycoproteins on the surface of bovine milk exosomes and intestinal cells facilitate the uptake of exosomes in human intestinal cells. The β‐galactoside, Core 1 & Core 3‐O‐linked disaccharide and N‐acetylglucosamine modifications appear to be of greater importance than other glycan features present on complex glycoproteins.Future DirectionsThe transport of bovine exosomes will be studied in conditional glycan knockout mice.Support or Funding InformationNIFA 2015‐67017‐23181, NIFA 2016‐67001‐25301/NIH DK107264, NIH 1P20GM104320, the Gerber Foundation, and USDA Hatch Act and W3002.
Association of Added Sugars Intake with Physiologic Parameters in Adults: NHANES 2001–2012
Authors: O'Neil Carol E , Nicklas Theresa A , Fulgoni Victor L 
Published: June 2021

The association of added sugars intake with liver enzymes, cardiovascular risk factors, and markers of red cell counts was determined using NHANES 2001–2012 data from adults (19+ years; n=26,402). Dietary intake was determined using 24‐hour dietary recall interviews using an Automated Multiple‐Pass Method. The usual intake of added sugars as a percent of energy was estimated using the Markov Chain Monte Carlo (MCMC) ratio method as implemented in v1.0 of the SAS programs published by the National Cancer Institute (NCI). Balanced repeated replication (BRR) was used for variance estimation. Subjects were separated into six groups: 0<5, 5 to <10, 10 to<15, 15 to <20, 20 to <25, and ≥25% of energy as added sugars. Group trends and linear trends for were determined using linear regression (p<0.01) for: alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, gamma‐glutamyl transferase, lactate dehydrogenase, blood pressure, high‐ and low‐density lipoprotein cholesterol, triglycerides, glucose, c‐reactive protein, waist circumference, ferritin, serum and red blood cell folate, hemoglobin, and hematocrit. None of the liver enzyme levels showed any significant association with added sugars intake. Mean plasma glucose levels (mg/dL) showed an inverse group (beta = −0.71; p=0.0003) and linear trend (beta = −0.13); p=0.0002) association with increasing percent energy of added sugars. No other cardiovascular risk factors, including metabolic syndrome, showed any association with added sugars intake. Only hemoglobin (g/dL) (beta = −0.03, p=0.0007; beta = 0.005, p=0.0013) and hematocrit (%) (beta = −0.09, p=0.0001; beta = −0.02, p=0.0002) showed a group or linear trend, respectively. No other statistical associations were found. These results suggest that the association of usual added sugars intake, as determined by sophisticated models, up to >25% of energy, had limited effect on physiologic parameters in adults.Support or Funding InformationUSDA Hatch Project LAB 94209 and USDA/ARS through specific cooperative agreement 58‐6250‐6‐003 was received. Partial support was also received from the Corn Refiners Association.
Neonatal diet composition modulates ileum mitochondrial function in a neonatal pig model
Authors: Carvalho Eugenia , Moreira Ana R , Bowlin Anne K , Saraf Manish K , Adams Sean H , Borsheim Elisabet , Yeruva Laxmi 
Published: June 2021

BackgroundThe composition of postnatal diet (i.e., breastmilk vs. formula) has a strong influence on a variety of physiological outcomes in infants, but the impact on bioenergetics and mitochondrial function remains an open question. In a published study (1), early ingestion of dairy‐based infant formula vs. human breast milk differently modulated liver mitochondrial function and gut microbiota in rodents. However, the effects on bioenergetics in other tissues, including the gut, and the persistent effect after weaning remain unknown. In the current study we used a piglet model to determine the effects of these two milk types on bioenergetics phenotypes in the ileum.MethodsNeonatal piglets were fed isocaloric diets of either human breast milk (n=9) or a dairy‐based infant formula (n=9) from 48 hours post‐delivery until day 21, followed by an ad libitum solid diet until day 51. Subsequently, ileum biopsies were collected for evaluation of permeabilized cell mitochondrial function by high resolution respirometry, following a substrate and inhibitor protocol. In addition, gene expression for a key regulator of mitogenesis and mitochondrial function, peroxisome proliferator‐activated receptor gamma coactivator 1‐alpha (PGC‐1α) was assessed.ResultsATP‐linked respiration tended to be increased in the ileum of piglets fed a dairy‐based infant formula compared to those fed with human breast milk, even at 4 weeks of post‐weaning neonatal diet (10.4 +/− 2.2 vs. 5.8 +/− 0.9 pmol·s−1·mg−1; p=0.07). Cytochrome c oxidase (COX) activity was significantly elevated in the formula compared to the human breast milk (23.3 +/− 4.8 vs. 9.9 +/− 1.8 pmol·s−1·mg−1; p=0.02). Thus, the substrate control ratio for succinate relative to COX was significantly lower in the dairy fed group (0.07 +/− 0.02 vs. 0.14 +/− 0.02; p=0.03). Interestingly, PGC‐1α gene expression tended to be elevated (2.3 fold, p=0.06) in ileum of formula‐fed piglets compared to those fed human breast milk.ConclusionWe conclude that different milk diets may have a sustained impact on ileum bioenergetics in neonatal piglets, even a month after discontinuation of the milk diets. Future efforts are needed to understand the implications of these novel findings on infant tissue function and whole‐body macronutrient metabolism.Support or Funding InformationFunded by USDA ARS Project 6026 51000 010 05S. FEDER and FEDER funds through the Operational Programme Competitiveness Factors ‐ COMPETE and UID/NEU/04539/2013.
Cytotoxicity of ethanol extracts from glanded and glandless cottonseed in cultured mouse RAW macrophages
Authors: Cao Heping , Sethumadhavan Kandan 
Published: June 2021

Cotton plant provides two economically important products, fiber as the major product and cotton seed as the minor product. However, cotton plant produces more seed in terms of quantity than fiber with a seed/fiber ratio of 1.5 – 1.7. Cottonseed typically accounts for 15 – 25% of the crop value. Due to presence of toxic gossypol in the commonly cultivated cotton (glanded cotton), the uses of cottonseed is limited to feeding cows and not for human and other animal consumption. To explore the potential use of glandless cottonseed which has only trace amount of gossypol, we investigated the cytotoxicity of extracts from glandless cottonseed along with those from glanded cottonseed and compared directly with gossypol and lipopolysaccharide (LPS) using cultured mouse macrophages. Cottonseeds were separated into coat and kernel fractions. Both fractions were homogenized in 0.1 N acetic acid, defatted with chloroform and hexane and extracted with ethanol. The ethanol extracts were rotoevaporated to remove ethanol and the dried extracts were reconstituted in DMSO before being added to cultured mouse RAW264.7 macrophages using gossypol and LPS as controls. Cell cytotoxicity was determined with MTT method. Dosage study showed that no cytotoxicity effect was observed in cells treated for 2–24 h with extracts (up to 1 mg/ml) from glandless cottonseed coat or kernels, nor with glanded cottonseed coat. However, mitochondrial activity measured by the MTT method was decreased by 40% with glanded cottonseed kernel extract after cells were treated with 0.1–1 mg/ml for 24 h. LPS (up to 1 ug/ml) did not show toxic activity but gossypol (≥5 ug/ml, 24h) almost completely inhibited cell viability. These results suggest that glandless cottonseed extractis harmless towards the cultured macrophages but glanded cottonseed extract from its kernel, like gossypol, is harmful to the immunological cells.Support or Funding InformationSupported by USDA‐ARS Quality and Utilization of Agricultural Products Research Program 306 through CRIS 6054‐41000‐103‐00D.
Singlet Oxygen Production in the Caudodorsal Medulla of Rat Brain Slices Increases During Hyperoxia at Normobaric Pressure and Following Decompression from Iso‐oxic Hyperbaric Nitrogen
Authors: Roberts Teryn R. , Ciarlone Geoffrey E. , Dean Jay B. 
Published: June 2021

Previous studies in brain slices harvested from the medulla oblongata of Sprague‐Dawley (SD) rats indicate that neurons in the caudal solitary complex (cSC) are stimulated by normobaric hyperoxia (NBO2) and hyperbaric O2 (HBO2). We've also reported that during NBO2, cSC cells produce a variety of reactive O2 and N2 species, including superoxide, nitric oxide, peroxynitrite, hydrogen peroxide, hydroxyl radicals, carbonate radicals, & nitrogen dioxide. These findings support the premise that cSC neurons employ redox signaling mechanisms that are activated by increasing pO2. Presumably, this explains, in part, the paradoxical stimulation of breathing by prolonged exposures to NBO2 and HBO2. We have tested the hypothesis that singlet oxygen (1O2) production increases in cSC cells exposed to a) NBO2 in the presence of nominal N2 and b) iso‐oxic hyperbaric nitrogen (HBN2). In the case of HBN2, we wanted to determine if inert gas molecules react with O2 molecules at pressure to form collision complexes that yield 1O2. The fluorogenic dye, Singlet Oxygen Sensor Green (SOSG, 2.5μM), was used to measure SOSG intensity of cSC neurons (P16–40 M/F SD rats, 400μm). Fluorescent images were captured at 1–2.5 min intervals and correlated with 1O2 production using one of two Nikon TE2000 inverted microscopes; one microscope was automated for use inside a hyperbaric chamber. The HCO3−‐buffered nutrient medium was equilibrated at room pressure with either (in atmospheres absolute, ATA) 0.4 O2 + 0.05 CO2 + 0.55 N2 (control) or 0.95 O2 + 0.05 CO2 + “0.0” N2 (NBO2).* For iso‐oxic HBN2, the air‐filled hyperbaric chamber (0.8 ATA N2) was compressed (0.25 ATA/min) with 100% N2 to 80–85 psig (5.4–5.8 ATA N2), thereby increasing atmospheric pN2 to 6.2–6.6 ATA (0.95 O2 + CO2 in the nutrient medium; pN2 increases in the medium and slice from HBN2 diffusing in from the overlying atmosphere). Nutrient media were delivered to the brain slice at 1.5 ml/min using an HPLC pump (30–31°C). Results show that increasing pO2 from 0.4 to 0.95 ATA at room pressure increased the rate of 1O2 fluorescence. 1–4 mM ascorbate, but not <1mM, decreased the rate of 1O2 fluorescence in 0.4 ATA O2 and NBO2 in a dose‐dependent manner. Exposure to iso‐oxic (0.95 ATA O2) HBN2 (10–30 min) did not increase 1O2; however, decompression (0.5ATA/min) back to sea level induced a post‐dive increase in 1O2 fluorescence that persisted for 10–30 min. The post‐dive increase in 1O2 fluorescence was inhibited by 4mM ascorbate. These findings suggest that 1O2 production is activated in cSC cells by hyperoxia, compression to at least 6.2 ATA N2, decompression, or the combination of compression/decompression in hyperoxic HBN2. We postulate that 1O2 generated in cSC neurons may contribute, in part, to CNS O2 toxicity, inert gas narcosis, and decompression sickness.Support or Funding InformationONR N000141512572 & N000141612537
The Growth and Differentiation Capacity of Porcine Renal Papilla Progenitor Cells Decline with Passaging
Authors: Burmeister David M. , McIntyre Matthew K. , Gomez Belinda I. , Mongomery Robbie , Dubick Michael A. 
Published: June 2021

When challenged by an ischemic insult, such as that accompanying traumatic hemorrhage, the kidney undergoes functional recovery via replacement of cells. Different theories about the cellular mechanisms behind this endogenous regeneration exist, however recent studies have identified a population of renal stem/progenitor cells from the renal papillae. With the increased use of swine in, for example, hemorrhagic shock models, we aimed to isolate and characterize progenitor cells from the renal papillae from pig kidneys. After excision and collagenase digestion of renal papillae, non‐adherent cells were removed after 2 days, and the ensuing renal progenitor cell (RPC) population was expanded under normal culture conditions. At passages 2 and 6, RPCs were examined for their: growth kinetics; expression of markers CD24, CD133, CD90, and NFATC1 via flow cytometry; and potential to differentiate into osteoblasts and adipocytes. Additionally, passage 2 RPCs were used in a mixed lymphocyte reaction to determine immunomodulatory activity. At passage 2, RPCs displayed a characteristic spindle‐shaped morphology with substantial proliferative capacity. By passage 6, accelerated vacuolization and minimal growth was seen in RPCs. Flow cytometry revealed that 96.3% of RPCs at passage 2 expressed CD24, which was dramatically reduced to 23.9% at passage 6. Expression of CD90 at the two passages was similar (44.4% and 47.2% at passage 2 and 6, respectively), while there was low expression (~10%) of CD133 or NFATC1 at both passages. Differentiation studies revealed that RPCs were able to differentiate into both adipocytes and osteoblasts at passage 2, but not at passage 6. Moreover, RPCs from passage 2 increased the amounts of IL‐6 and IL‐8 (p<0.05) released in the mixed lymphocyte reaction, but tended to reduce the amount of Interferon‐γ released. Taken together, these data further indicate that porcine RPCs are heterogeneous and lose their “stemness” properties with serial passaging. However, their immunomodulatory capabilities lend them the potential to be leveraged as therapies or therapeutic targets. In instances of ischemic insult, augmentation of RPCs may potentiate the recovery and regenerative properties of the kidney. Further work exploring the characteristics of renal stem cell populations, their interactions with resuscitation fluids, and their efficacy in pre‐clinical animal models is warranted.Support or Funding InformationUS Army Medical Research and Materiel Command (MRMC) provided funding for this project.
Balance of Conformational States Affect the Intrinsic Hydrolysis of NRas When Compared to Other Ras Isoforms
Authors: Reid Derion F , Mattos Carla 
Published: June 2021

Ras GTPases are small proteins that are involved in various signal transduction pathways that regulate cellular proliferation, survival, migration, and apoptosis. Although the first crystal structure of Ras was determined more than two decades ago, biochemical and structural differences in the G‐domain of the three isoforms, H‐, K‐ and N‐Ras is only now beginning to emerge. Each isoform functions as a molecular switch where guanine nucleotide exchange factors (GEFs) facilitate the exchange of guanosine diphosphate (GDP) for guanosine triphosphate (GTP) turning its signal on. Ras signaling is turned off via intrinsic hydrolysis or through the aid of GTPase activating proteins (GAP). Ras mutants are involved in 20% of human cancers, yet there are no inhibitors that target these proteins effectively. Most of the effort was originally focused on HRas as the classical representative of all three isoforms and more recently on KRas, the most frequently mutated in cancers. Thus, the structural biology and biochemistry of NRas have been largely overlooked. We have determined hydrolysis rate constants for NRas and determined the crystal structure of wild‐type NRas bound to GppNHp (GNP), a non‐hydrolyzable analogue of GTP, for comparison with H‐Ras and K‐Ras. Although each of the three isoforms follow a one‐step mechanism of hydrolysis, K‐ and NRas have lower rates than HRas, with structural features that promote disorder in switch II near the active site. N‐Ras mutants at G12 and Q61 are commonly found in melanoma tumors. These conserved residues are found in the phosphate‐binding loop (G12) and switch II (Q61) that help make up the active site of NRas. The structures of NRas mutants reveal distinct mechanisms through which they affect catalysis, consistent with lower hydrolysis rate constants found for G12D, G12S, Q61H, and Q61K compared to wild‐type NRas. Biochemical and structural understanding of NRas mutants will help further influence the development of novel therapeutic targets of NRas mutant melanoma.Support or Funding InformationThis research was funded by NSF Grant Number MCB‐1517295.
A High Serum Vanin‐1 Phenotype is Not Unique to Diving Marine Mammals
Authors: Boxall Baylye K. , Prager Katherine , Neely Benjamin A. , Lloyd‐Smith James O. , Gulland Frances , Janech Michael G. 
Published: June 2021

Diving marine mammals have elevated levels of antioxidant proteins in their tissues to counter oxidative stress in part due to diving. A proteomic study of bottlenose dolphin serum identified and measured the highest concentration of pantetheinase (Vanin‐1) for any mammal. Vanin‐1 is known to produce the antioxidant, cysteamine, from the vitamin B5 precursor, pantetheine. High levels of this protein may represent an adaptation or response to diving in marine mammals. If this is true, then elevated levels of circulating Vanin‐1 may be a state shared amongst, but unique to, diving marine mammals. To test this hypothesis, sera from a diving pinniped (N=6), California sea lion (Zalophus californianus), were assayed for Vanin‐1 concentration by liquid chromatography/tandem mass spectrometry using parallel reaction monitoring. Commercially purchased sera from 9 terrestrial species belonging to five different mammalian orders: Rodentia (Muridae), Primates (Hominidae), Cetartiodactyla (Suidae and Bovidae), Perissodactyla (Equidae), and Carnivora (Canidae) were also monitored via product ion scan for a tryptic peptide homologous to the sea lion Vanin‐1 peptide. The presence of the monitored Vanin‐1 peptide was detected in 5/6 sea lion samples as defined by a lower limit of detection of 3.7 μg/mL, which was calculated by 3.3σ of the lowest standard used in an external calibration curve. Only 3/6 serum samples had measureable concentrations above the lower limit of quantification of 5 μg/mL, which was calculated by 10σ of the lowest standard. Of those samples above the lower limit of quantification the mean ± standard deviation of Vanin‐1 concentration was 5.7 μg/mL ±0.4. Monitoring of serum Vanin‐1 by product ion scan in terrestrial mammals revealed a signal that was well above the lower limit of quantification in sheep, cow, goat, donkey, and horse, but not detected in pig or dog. Product ion scan data in these species were consistent with proteomic rank data. These findings indicate that a high circulating Vanin‐1 phenotype is not unique to diving marine mammals. Data suggest that an elevated Vanin‐1 phenotype is shared amongst some members of the clade Ungulata, which includes the orders Perissodactyla and Artiodactyla, within which fall cetaceans.Support or Funding InformationResearch was supported in part by the Office of Naval Research grant N000141410361, National Science Foundation grant OCE‐1335657, and the John H. Prescott Marine Mammal Rescue Assistance Grant Program.