bluuebunny/crossref_metadata_2025 huggingface dataset:
Behavior related genes, dietary preferences and anthropometric traits
Authors: Berciano Silvia , Lai Chao‐Qiang , Herranz Jesus , Aslibekyan Stella , Claas Steve A , Irvin Marguerite R , Tsai Michael Y , Hopkins Paul N , Hidalgo Bertha , Arnett Donna K , Ordovas Jose M , Tiwari Hemant
Published: June 2021

Food preferences and cognitive control influence dietary habits thus affecting compliance with dietary advice and the risk of chronic diseases. A complex mix of genetic and environmental, cultural and social factors drives these preferences. Therefore, efforts to improve diet and behavior at the individual level should take into consideration this distinct combination of genetic and environmental factors, specifically addressing the psychological component of food consumption with the goal of facilitating long‐term compliance with dietary interventions.

Objective

To investigate the association between behavioral candidate genes, food preferences and anthropometric traits.

Population and Methods

For this purpose we used data from the Genetics and Lipid Lowering Drugs and Diet Network (GOLDN) Study including two genetically homogeneous sites (Minneapolis, Minnesota, and Salt Lake City, Utah). In the current study, 818 participants (404 men and 414 women) of European ancestry were included in our analyses. Single nucleotide polymorphisms (SNPs) within 38 loci (1359 SNPs) selected on the basis of previous associations with several behavioral and psychological traits (i.e., stress, addiction, depression, impulsivity, novelty‐seeking, aberrant eating) were extracted from the original genome genotype data that was generated from the Genome‐Wide Human SNP Array 6.0 (Affymetrix). Information about dietary intake was collected with the use of a diet‐history questionnaire, which was developed by the National Cancer Institute.

Results

Multiple nominally significant associations (p<0.05) were observed between genetic variability at the selected loci and the consumption of specific foods and nutrients. However, after adjustment for multiple comparisons, significant associations (Padj<0.05) were observed for the FTO locus with vegetable and total fiber intake; the CREB1 and GABRA2 loci were associated with salt intake; and the SLC6A2 with total fat and monounsaturated fatty acids. Finally, chocolate intake was associated with variation at the OXTR locus. The most significant association with anthropometric traits was found for OXTR and waist circumference.

Conclusion

Our data indicate that genes implicated in behavioral and psychological traits drive a significant component of an individual's food preferences and dietary habits. This information will contribute to a better understanding of eating behavior and facilitate the implementation of personalized dietary advice that should result in better compliance and more successful prevention and therapy of chronic disorders.

Support or Funding Information

National Heart, Lung, and Blood Institute (NHLBI) grants U01HL072524 (Genetic and Environmental Determinants of Triglycerides), NHLBI R01 HL091357 and by contracts 53‐K06‐5‐10 and 58‐1950‐9‐001 from the USDA, Agriculture Research Service
A Genome‐Wide Analysis of Placental Epigenetic Markers in Response to Maternal Choline Supplementation
Authors: Kwan Sze Ting (Cecilia) , Wang Minghui , King Julia H , Yan Jian , Jiang Xinyin , Sun Qi , Roberson Mark S , Caudill Marie A
Published: June 2021

Choline is an essential micronutrient that provides methyl groups for cellular DNA methylation, an epigenetic modification with downstream effects on gene expression. Maternal choline supplementation (MCS) during pregnancy has many beneficial effects on placental and fetal development, which may be mediated in part by an altered placental methylation profile. In addition to DNA methylation, other epigenetic mechanisms such as genomic imprinting and microRNA (miRNA) regulation are also important to normal placental and fetal development, but whether these markers are modulated by choline remains largely unknown. We sought to investigate the impact of MCS on these epigenetic markers using a subset of placental samples collected at gestational day 15.5 from dams consuming 1× or 4× the recommended choline level. Placental mRNA and miRNA were extracted and sequenced, and the RNAs with differential expression were determined using the edgeR package developed in R. In response to MCS, three imprinted genes (Dcn, Qpct and Tnfrsf23) were downregulated in the male placentas (Padj≤0.01) whereas four imprinted genes (Ampd3, Tfpi2, Gatm, and Aqp1) were upregulated in the female placentas (Padj≤0.05). Furthermore, five miRNAs (miR‐712‐5p, miR‐3470a, miR‐6538, miR‐6240 and miR‐5126) were significantly downregulated in the female placentas from the 4× choline group (Padj≤0.033). Consistent with the downregulation of these miRNAs, some of their predicted mRNA targets showed a higher expression (P<0.05). These target genes are involved in processes such as cell signaling, immune response and macronutrient transport, all of which are crucial to placental development and fetal health. In contrast, none of the miRNAs in the male placentas remained significant after correction for false discovery rate (Padj>0.05). Overall, these data indicate that the placental imprintome and miRNA profile are responsive to MCS, but the effects may differ between the male and the female placentas.

Support or Funding Information

USDA, NIH, Graduate Women in Science, Egg Nutrition Center
Reducing Added Sugar Intake Increases the Relative Reinforcing Value of High‐Sugar Foods
Authors: Flack Kyle David , Roemmich James , Johnson LuAnn
Published: June 2021

Objective

To determine whether reducing added sugar intake to <10% of calories for 1 week changes the relative reinforcing value (RRV) of foods high in sugar in non‐overweight and obese adults.

Background

The 2015–2020 Dietary Guidelines of Americans (DGA) focuses on 5 overarching guidelines for healthy eating patterns, including limiting calories from added sugars to less than 10% of calories/day. Foods high in added sugars (e.g., soda, desserts, candy) are reinforcing and reducing their consumption in order to adhere to the DGA may paradoxically increase their RRV.

Methods

Twenty seven adults aged 18–39 y with a BMI below 25 kg/m2 (n=18) or above 30 kg/m2 (n=9) who reported consuming >10% of daily calories from added sugar completed a 1‐week controlled feeding intervention. Participants consumed ~3% of their daily calories from added sugar while maintaining energy balance and the average macronutrient distribution for adults over the age of 20 (~16% En pro, 51% En CHO, 33% En fat). Prior to and on the seventh day of the dietary intervention participants completed an operant responding task that used independent progressive ratio schedules of reinforcement to earn a high‐added (e.g. skittles, tootsie roll) and a low‐added (e.g. potato chips, pretzels) sugar food. The RRV of added sugar foods was calculated as the number of sessions completed for access to a high‐added sugar food/(sessions completed for high‐added sugar food + sessions for low‐added sugar food). Hunger and satiety were measured prior, and calories consumed of the earned food measured after, RRV tests.

Results

RRV of high‐added sugar foods increased (47%, p<0.001) from 0.51 at baseline to 0.77 post with no significant effects of weight status, hunger or satiety on RRV change. All but 2 participants earned high added sugar food Pre, but all earned high‐added sugar foods Post (p = 0.15). For low‐added sugar foods, all but 2 participants earned some Pre, but only 14 of 27 earned any Post (p < 0.001).

Conclusions

In adults, the RRV of foods high in added sugar increases when access to these foods is reduced for 7 days below their base rate to meet the DGA. This presents a potential mechanistic barrier to meeting the DGA recommendations for reducing added sugar and why many Americans have difficulty making food choices that promote adherence to a reduced added sugar intake diet.

Support or Funding Information

This study was supported by USDA‐ARS Project USDA‐ARS 3062‐51000‐051‐00D
Diversity of the primate musculoskeletal system: modularity, integration, and anatomical variation in humans and their relatives
Authors: Molnar Julia L
Published: June 2021

The concepts of modularity and integration help to explain how morphological transformations during primate evolution enabled modern humans to carry out complex functions such as speaking, chewing, and using tools while also being able to respond to selective pressures. We used anatomical network analysis (AnNA) to assess modularity and integration in the head and upper limb of extant primates. AnNA is the study of complex morphological structures using tools and statistics from network theory. In this context, a module is a group of anatomical elements (e.g., bones and muscles) with more connections (i.e., more integrated) among them than to any other elements outside the group. These modules are identified using community detection algorithms. We analyzed the networks of the head and upper limbs of 20 taxa representing all the major extant primate groups based on dissection of hundreds of specimens, observation of skeletons, and an extensive literature review to ensure that the total sample sizes per taxon would be sufficient to determine the normal muscle phenotype for each taxon. Our results reveal patterns of evolutionary change that are highly relevant for biological and physical anthropology and primatology. For example, the division into left and right orofacial muscle modules seen in humans is not found in bonobos or common chimps, meaning that this division may be related to the more complex, asymmetrical facial expressions displayed by humans.

Support or Funding Information

NSF grant 1516557 (2015–16) to Rui Diogo and American Association of Anatomists Innovations Program
Sexual Dimorphism in Bones of the Thenar and Hypothenar Aspects of the Hand
Authors: Williams Avery A. , Patel Biren A. , Tocheri Matthew W. , Orr Caley M.
Published: June 2021

Osteoarthritis of the pollical carpometacarpal joint is significantly more prevalent in females than in males. Although a number of hypotheses to explain sex differences in the prevalence of thumb osteoarthritis have been proposed, one possibility is that it is a product of sexual dimorphism where female hands have smaller joint surfaces relative to muscle strength, which results in higher stresses on the joints. Although dimorphism of the skull and long bones has been extensively researched, there are few basic data on female‐versus‐male differences in the short and irregular bones. Therefore, it is unclear whether sexual dimorphism is a relevant avenue of investigation for determining the cause of such sex‐based differences in hand pathology. Because powerful precision gripping is dependent on the thenar and hypothenar muscles, the hand bones with muscle attachments or that are directly subjected to transarticular loading by these muscles are of particular interest. Therefore, we investigated four hand bones contributing to the first and fifth carpometacarpal articulations in a sample of humans of African and European descent, and it was predicted that females would have smaller overall bone volumes and areas, smaller individual joint surface areas, as well as different joint and muscle attachment shapes. The hamate, trapezium, first metacarpal (Mc1), and fifth metacarpal (Mc5) of each individual were laser scanned to generate 3D surface polygon models. Volume and surface area were calculated for each bone as a whole in addition to the areas of specific joint surfaces. The shape of the pollical carpometacarpal joint (proximal Mc1 and distal trapezium) was assessed with quadric fitting functions to determine their dorsovolar and radioulnar curvatures. For the hamate, the orientation of the triquetrohamate joint surface relative to the proximodistal axis was quantified, and for the hamulus (which provides attachments for the flexor digiti minimi brevis, opponens digiti minimi, and flexor carpi ulnaris) its length, width, breadth, and angle were measured. The results indicate that females tend to have bones that are significantly smaller in overall size with correspondingly smaller joint surfaces, and a hamulus that differs in both size and shape. Future work will diversify the sample and incorporate measurements of relative thenar and hypothenar muscle force potential to further evaluate the biomechanical consequences of skeletal dimorphism. Such an approach may allow us to better investigate more specific factors contributing to the increased osteoarthritis risk in the hands of females.

Support or Funding Information

National Science Foundation (BCS‐1539741, BCS‐1317047), Wenner‐Gren Foundation, Leakey Foundation
A novel, high precision multiple‐meal stable isotope method to compare iron absorption from extruded FePP‐fortified rice containing different zinc compounds, citric acid/trisodium citrate and EDTA in Ghanaian children
Authors: Hackl Laura , Abizari Abdul‐Razak , Zungbey‐Garti Helene , Zeder Christophe , Speich Cornelia , Cercamondi Colin I. , Zimmermann Michael B. , Moretti Diego
Published: June 2021

Rice fortification can be a viable approach to combat iron deficiency in rice‐consuming populations, but it is crucial to identify micronutrient formulations with high iron bioavailability and acceptable sensory properties. To date, ferric phosphates are the only iron compounds resulting in sensory acceptable iron fortified rice grains.

We measured fractional iron absorption (FAFe) from isotopically labeled ferric‐pyrophosphate (54FePP, 57FePP, 58FePP) in a cross‐over multiple meal absorption study. Fortified extruded rice meals either contained: zinc oxide (ZnO; 54FePP+ZnO), zinc sulphate (ZnSO4; 57FePP+ZnSO4), alone or in combination with a citrate buffer CA/TSC (54FePP+ZnO+CA/TSC or 57FePP+ZnSO4+CA/TSC) or ZnO, CA and edetate (EDTA; 58FePP+ZnO+CA+EDTA).

Iron depleted school‐age children with and without anemia (N=26) in Northern Ghana were fed six different rice meals (all meals containing 2mg iron) over the course of six weeks. Each type of meal was administered twice daily for five consecutive days. FAFe was compared from meals 54FePP+ZnO, 57FePP+ZnSO4, 54FePP+ZnO+CA/TSC, 57FePP+ZnSO4+CA/TSC, 58FePP+ZnO+CA+EDTA versus non‐fortified extruded rice with 58FeSO4 (reference) added after cooking. FAFe was measured as erythrocyte‐incorporation of stable iron isotopes at least 11 days after meal‐administration.

Geometric mean FAFe (95% CI) from meals 54FePP+ZnSO4+CA/TSC (6.3%; 5.3,7.4) and 58FePP+ZnO+CA+EDTA (6.5%; 5.3,8.1) did not differ from the reference (6.6%; 5.4,8.1). FAFe between 57FePP+ZnSO4 (3.5%; 2.7,4.5) and 54FePP+ZnO+CA/TSC (4.4%; 3.6,5.5) was not different, but both differed from 54FePP+ZnSO4+CA/TSC, 58FePP+ZnO+CA+EDTA and the reference (P<0.038), however, 54FePP+ZnO (2.3%; 1.9,2.8) showed the lowest FAFe and significantly differed from all other meals (P<.035).

Conclusions

Iron absorption from FePP‐fortified rice is affected by both the zinc source (ZnO or ZnSO4) and absorption enhancers (CA/TSC and CA+EDTA). For maximal iron absorption, rice fortification should be implemented using ZnSO4 as the zinc fortificant and CA/TSC or CA+EDTA as iron absorption enhancers.

Support or Funding Information

This study was funded by the Laboratory of Human Nutrition, DSM Nutritional Products, USAID and Abbott Nutrition.
Ablation of systemic SIRT1 activity promotes nonalcoholic fatty liver disease by affecting liver‐mesenteric adipose tissue fatty acid mobilization
Authors: Cheng Junrui , Liu Chun , Hu Kangquan , Greenberg Andrew , Wu Dayong , Ausman Lynne M. , McBurney Michael W. , Wang Xiang‐dong
Published: June 2021

The incidence of nonalcoholic fatty liver disease (NAFLD) is escalating paralleled with obesity rates in both adults and children. Mammalian sirtuin 1 (SIRT1), a highly conserved NAD+‐dependent protein deacetylase, has been identified as a metabolic regulator of lipid homeostasis and a potential target for NAFLD prevention and treatment. However, the mechanism of how SIRT1, in particularly its deacetylase activity, affects NAFLD has not been well investigated. The current investigation addressed the causal effect of systemic SIRT1 activity on NAFLD development and the potential mechanism involved in both liver and mesenteric adipose tissue. At 6 weeks of age, both SIRT1 homozygous mice carrying a point mutation (H355Y, SIRT1Y/Y, n=10) that ablates the catalytic activity and their corresponding wild type littermates (WT, n=10) were fed with high fat diet (HFD, 60% calories from fat) for 34 weeks. Results showed no changes of body weight, liver weight and body composition (anlyzed by EchoMRI‐700) between WT and SIRT1Y/Y mice, but SIRT1Y/Y mice showed significantly higher level of hepatic triglyceride, and a trend of more severe hepatic steatosis by pathological analysis, which was accompanied with higher protein levels of LXRα, SREBP‐1, SCD1, involved in lipogenesis, and decreased phosphorylation of LKB1 and AMPK in the liver. In the mesenteric adipose tissue, mRNA expression of lipogenic genes (lxrα, srebp‐1c, scd1 and fas) increased significantly in SIRT1Y/Y mice, as compared with those of wild‐type mice. Fatty acid oxidation biomarkers (acox1, acox3, cpt, ucp1, sirt3) in both liver and mesenteric adipose tissue were comparable between groups. Interestingly, we observed that in SIRT1Y/Y mice, the mRNA level of hormone sensitive lipase (hsl), adipose triglyceride lipase (atgl) and perilipin‐2, involved in lipolysis, significantly increased in mesenteric adipose tissue (not in epididymal adipose tissue), as compared with those of WT. These changes were correlated with higher circulating free fatty acid (FFA) concentrations and higher hepatic mRNA expression of cd36 for FFA uptake. Taken together, the present study provided novel experimental evidence that under HFD‐induced metabolic surplus, the lack of SIRT1 catalytic activity promoted release of FFA from mesenteric fat into systemic circulation and escalated NAFLD by interfering with lipid homeostasis in both liver and mesenteric adipose tissue.

Support or Funding Information

Supported by USDA/NIFA (2015‐67017) and USDA/ARS (58‐1950‐0014)
Introduction of Enzymatic Polymer Degradation to enhance the Undergraduate Polymer Chemistry Curriculum
Authors: Neely Amy E , Davis Nicole Y , Weiland Mitch H
Published: June 2021

Changes to the 2015 ACS Guidelines and Evaluation Procedures for Bachelor's Degree Programs included mandatory instruction in polymers and macromolecules for the ACS‐certified degrees. To satisfy this mandate, we have begun developing a series of undergraduate laboratory exercises involving the synthesis, degradation, and characterization of poly(aspartic acid) (PAA). PAA, a biodegradable, water‐soluble synthetic polymer, could potentially replace commonly used poly(acrylic acid), which is a non‐biodegradable polymer that possess environmental risks. In this module of experiments, PAA is prepared in the organic chemistry laboratory course by polymerizing aspartic acid to form poly(succinimide) as an intermediate, which is converted to PAA by base induced ring‐opening. The synthetic PAA, composed of both β‐amide (70%) and α‐amide units (30%), is then transferred to the biochemistry laboratory course for enzymatic biodegradation using two enzymes, PAA hydrolase‐1 (PAAH‐1) and PAA hydrolase‐2 (PAAH‐2). A proposed mechanism suggests that PAAH‐1 hydrolyzes β‐β amide linkages, resulting in oligo (aspartic acid), wherein PAAH‐2 completes the degradation process by cleaving the oligo (aspartic acid) units into monomeric aspartic acid. Current progress on the development of this biochemistry laboratory exercise including overexpression of recombinant PAAH‐1 and PAAH‐2, purification, and characterization will be presented.

Support or Funding Information

DUE 1611988
Profiling of DNA methylation patterns in response to inorganic arsenic exposure and reversal of exposure
Authors: Eckstein Meredith Lee , Rea Matthew , Eleazer Rebekah , Fondufe‐Mittendorf Yvonne
Published: June 2021

Epigenetic modifications are heritable changes in gene expression not encoded in the DNA sequence. One such epigenetic mark that has been extensively studied is DNA methylation as it is most stable and also reversible. Recent studies suggest that epigenetic regulation through DNA methylation could be responsible for cellular transformation instigated by environmental toxins, such as inorganic arsenic. Research has been done to investigate DNA methylation and gene expression patterns at specific genes during this process; but so far, a comprehensive understanding of the impact of DNA methylation changes on gene expression patterns is missing. This study presents a comprehensive genome‐wide DNA methylation analysis in response to chronic, low‐dose inorganic arsenic exposure and the removal of this toxin. We aim to determine how inorganic arsenic exposure leads to the epithelial‐to‐mesenchymal transition in cellular transformation and to understand mechanistically the importance of DNA methylation in driving this process. We show that several key genes are differentially expressed during this process and that some of these expression states revert when inorganic arsenic is removed. To understand the functional role of DNA methylation in this process, we profiled DNA methylation changes at individual CpG loci using the Infinium Methylation EPIC Bead Chip Methylation array. We found that chronically inorganic arsenic‐exposed cells showed changes in DNA methylation corresponding with changes in their expression profiles. Many of the methylation changes were concentrated at upstream promoter regions and gene regulatory regions, suggesting a functional relevance in inorganic arsenic‐mediated cellular transformation. Our studies indicate that inorganic arsenic‐mediated transformation is instigated, in part, by DNA methylation pattern changes that cause alterations in gene expression and reversal of inorganic arsenic treatment moderately reverses the methylation and gene expression changes.

Support or Funding Information

NSF (MCB 1517986‐YFM), NIEHS (R01‐ES024478‐YFM)
Acetaminophen Hepatotoxicity testing using 3D rat hepatocyte cultures
Authors: Paliwal Vipin , Clapham Margaret
Published: June 2021

Animal models are currently used for drug development and testing but are expensive and often inaccurate. Monolayer (2D) cell culture models have been utilized due to lower cost, but have proven less reliable because they are unable to preserve the functionality of the liver in vitro. This study aimed to establish a more accurate and inexpensive pharmaceutical testing method that has reliability and reproducibility. H35 Rat Reuber hepatoma cells were grown in both two‐dimensional (2D) and three‐dimensional (3D) cell cultures. Cell cultures were treated with different concentrations of acetaminophen (APAP) and analyzed quantitatively for cytotoxicity using a Neutral Red Dye. Cell function was then assessed by quantification of albumin secretion using ELISA. Results showed that both 2D and 3D cultures declined in albumin production after 48 hrs treatment with 2.25 mM APAP treatment. After 72 hrs of treatment, 3D cells showed an 81.2% decline in albumin secretion even with a much lower dose of 0.15 mM APAP whereas 2D cells in fact increased albumin secretion slightly by 0.63% at this concentration.

We conclude that when cells are grown in 3D cultures, an environment physiologically more similar to that of the liver, they have much higher sensitivity to acetaminophen hepatotoxicity.

Support or Funding Information

1) National Science Foundation, 2) Milwaukee School of Engineering and 3) Medical College of Wisconsin
Peptide Facilitated Intracellular Delivery of Quantum Dots for Live‐Cell Imaging
Authors: Vera Claudia Ivette Rivera
Published: June 2021

Semiconductor Quantum Dots (QDs) are luminescent colloidal nanocrystals that display unique optical properties resulting from a combination of their core semiconductor materials and their nanometer‐sized quantum confined effects. QDs are composed of semiconductor cores, which are often coated with one or more shell(s) consisting of another semi‐conductor material with suitable lattice parameters and higher band gap energy (for type I QDs; e.g., CdSe/ZnS core/shell or CdSe/CdS/ZnS core/shell/shell QDs). The photophysical properties of QDs can be controlled by their core sizes, the shell thickness, the composition of the cores/shell(s), and partly by their surface ligands. Thus, the final absorption and emission features of QDs can be tuned from the UV to the near‐IR to fit the desired spectral characteristics, making them ideally suited for many spectroscopic and imaging applications. Due to their outstanding photophysical properties, QDs are suited for a wide range of applications as labels and sensors in biochemistry, biotechnology and medicine (e.g. fluorescence spectroscopy and microscopy as well as FRET‐based assays and diagnostics).

In order to create stable, bio‐functional QDs, hydrophobic surface capping ligands are either exchanged with or converted to hydrophilic ligands that interact with the QD shell surface (e.g., amphiphilic polymers, dithiols) and provide a surface layer that affords aqueous solubility and allows further conjugation to proteins, DNA or other biomolecules, for example, The Medintz group at Naval Research Laboratories (NRL) has developed several types of QD‐functionalizing ligands that offer a variety of chemistries for bioconjugation.

In this study we aim to passively deliver QDs‐bioconjugates to the cytosol of cells for subsequent live cell imaging experiments, the results will be discussed.

Support or Funding Information

NSF
Tracking the flow of carbon toward oil synthesis in an emerging crop lesquerella (Physaria fendleri)
Authors: Cocuron Jean Christophe , Tsogtbaatar Enkhtuul , Alonso Ana Paula
Published: June 2021

Lesquerella (Physaria fendleri) is a Brassicaceae that produces hydroxy fatty acids (HFAs) in its embryos; a type of oil that is very valuable and widely used in the industry of cosmetics, coatings, greases, plastics, paints, and biofuel, among others. The current source of HFAs is ricinoleic acid which comes from castor (Ricinus communis). Unfortunately, its large‐scale production has been banned from the United States because castor contains the highly toxic compound ricin. On the other hand, free of toxins and rich in HFAs, lesquerella is an attractive alternative to imported castor oil, and is hence in the verge of commercialization. Moreover, lesquerella has tremendous potentials for oil production, has a short maturity time and is not used for food compared to other oilseed crops. However, to become economically viable, lesquerella first must be engineered to synthesize more HFAs. A lack of knowledge of the metabolic pathways underlying oil synthesis in lesquerella embryos presents a major constraint.

In plants, fatty acid synthesis occurs in plastids and requires carbon in the form of acetyl‐CoA units as well as reducing power and energy. In order to assess the contribution of the pathways involved in fatty acid biosynthesis in terms of carbon, reductant and energy, a 13C‐Metabolic Flux Analysis, aka fluxomics, is being conducted. 13C‐labeling of developing embryos is required for this purpose. Our most recent work done towards meeting this goal is: (i) obtaining culture conditions for growing lesquerella embryos in vivo so that their biomass composition matches that of embryos grown in planta, (ii) determining the efficiency at which the embryos convert carbon into biomass (iii) tracking the flow of carbon towards oil synthesis using labeled substrates (13C‐glucose or 13C‐glutamine). The combination of the data from the above studies will permit to construct a flux map that will unravel possible “road blocks” in fatty acid production. Future work will consist in targeting these limiting steps through metabolic engineering to increase total fatty acids in lesquerella embryos.

Support or Funding Information

Agriculture and Food Research Initiative competitive grant # 2016‐67013‐24605 of the USDA National Institute of Food and Agriculture.
Probing the lipid composition at the site of influenza virus assembly and budding with high‐resolution SIMS
Authors: Kraft Mary L. , Yeager Ashley N. , Weber Peter K. , Zimmerberg Joshua
Published: June 2021

The formation of infectious influenza virions is sensitive to the cholesterol levels in the host cell. The findings that the influenza viral envelope is enriched with cholesterol and sphingolipids and cholesterol depletion alters the clustering of influenza virus proteins in the plasma membrane, which is detrimental to virus assembly, suggests that the influenza virus assembles and bud from cholesterol‐ and sphingolipid‐enriched domains in the plasma membrane. Although this hypothesis has been around for decades, whether the region of the plasma membrane from which the influenza virus buds is enriched with cholesterol and sphingolipids is still unknown. To answer this question, we have used high‐resolution secondary ion mass spectrometry (SIMS) to image the distributions of stable isotope‐labeled cholesterol and immunolabeled influenza virus protein in the plasma membrane during virus budding with ~100 nm lateral resolution. This work involved metabolically incorporating distinct stable isotopes into the cholesterol and lipids of interest in living MDCK cells. After infecting these MDCK cells with influenza virus for 24 h, we immunolabeled the influenza envelope protein, hemagglutinin. Then we imaged the isotope‐labeled cholesterol, lipids, and immunolabeled influenza hemagglutinin on the surfaces of the virus infected cells with ~100 nm lateral resolution. The resulting images clearly show that the influenza virus does bud from cholesterol‐enriched domains in the membranes of MDCK cells.

Support or Funding Information

This research was supported by the U.S. National Science Foundation under CHE 15‐08662.
A Prokaryotic‐like Lysophosphatidic Acid Acyltransferase Reveals Unique Features of Triacylglycerol Biosynthesis in Microalgae
Authors: Kim Yeongho , Terng Ee Leng , Riekhof Wayne , Cahoon Edgar B. , Cerutti Heriberto
Published: June 2021

Understanding algal triacylglycerol (TAG) metabolism has strong implications for the biology of storage lipids as well as for biofuel production. Higher plants have prokaryotic (chloroplast located) and eukaryotic (endoplasmic reticulum located) pathways to synthesize specific glycerolipids. Lysophosphatidic acid acyltransferases (LPAATs) in the chloroplast prefer adding C16 fatty acids at the sn‐2 position of the glycerol backbone, generating ‘prokaryotic’ lipid precursors. In contrast, LPAATs in the endoplasmic reticulum (ER) direct the preferential esterification of C18 fatty acids at the sn‐2 position, synthesizing ‘eukaryotic’ lipid precursors. In higher plants, triacylglycerols are considered eukaryotic lipids, as they have predominantly C18 esterified at the sn‐2 position. Moreover, TAG assembly enzymes have been found to localize to the ER in diverse eukaryotes.

Chlamydomonas reinhardtii, a unicellular green alga, has primarily C16 fatty acids esterified at the sn‐2 position of TAGs. This observation led to the proposal that Chlamydomonas mainly employs the prokaryotic (chloroplastic) pathway for TAG precursor biosynthesis. However, an alternative hypothesis is suggested by our characterization of a unique C. reinhardtii LPAAT, termed CrLPAAT2. In phylogenetic analyses this enzyme belongs within a Chlorophyte‐specific clade (limited to the Chlorophyceae and Trebouxiophyceae classes of green algae) and it is absent from all other eukaryotes examined. Interestingly, CrLPAAT2 displays a chimeric structure between known eukaryotic and prokaryotic LPAATs and it was found to localize to the ER. RNA‐mediated silencing of CrLPAAT2 indicated that the enzyme is required for TAG accumulation under nitrogen deprivation. We also demonstrated that CrLPAAT2 prefers as substrate C16:0‐CoA over C18:1‐CoA in vitro, by substrate competition assays, and in vivo, by complementation of a LPAAT‐defective yeast strain. Thus, Chlamydomonas (and related green algae) possesses a distinct glycerolipid synthesis pathway that relies on CrLPAAT2 to generate prokaryotic‐like TAG species (with C16 at the sn‐2 position) in the endoplasmic reticulum.

Support or Funding Information

This research is funded by EPSCoR (1004094).
Retention of provitamin A carotenoids in high‐provitamin A biofortified maize and eggs after various cooking methods
Authors: Sowa Margaret , Goltz Shellen , Tanumihardjo Sherry A.
Published: June 2021

Biofortification of staple crops to increase the content of provitamin A carotenoids (such as beta‐carotene and beta‐cryptoxanthin) is a strategy being implemented to increase dietary intake of vitamin A in regions where deficiency presents a widespread problem. Heat and light can cause isomerization and oxidation of carotenoids, potentially reducing the provitamin A activity of foods made with biofortified maize after cooking and processing. Thus, an understanding of the provitamin A retention after cooking is important for assessing the utility and maximizing the efficacy of biofortified foods. Retention of carotenoids in eggs from chickens fed high‐beta‐cryptoxanthin maize was assessed after frying, boiling, and microwaving. Retention of carotenoids in high‐beta‐cryptoxanthin maize was assessed in muffins, tortillas, porridge, and fried polenta made from both whole‐grain and sifted maize flour. This work demonstrates that provitamin A retention may be influenced by cooking, and processing methods should be taken into consideration when breeding and implementing biofortified maize varieties.

Support or Funding Information

Supported by USDA Hatch WIS01804, Global Health Funds at UW‐Madison, and Harvest Plus breeding contract 5204.
Prolonged High Altitude Exposure Results in Elevated Erythroferrone and Diminished Hepcidin Levels in Healthy Young Male Volunteers
Authors: McClung James P , Hennigar Stephen R. , Berryman Claire E. , Young Andrew J. , Pasiakos Stefan M.
Published: June 2021

Iron, an essential micronutrient, supports physical and cognitive performance through the binding of proteins and enzymes involved in oxygen transport and energy metabolism. High altitude (HA) acclimatization affects iron metabolism and results in increased hemoglobin concentrations due to hypoxia‐stimulated hemoconcentration and erythropoeisis. Iron homeostasis is regulated by hepcidin, aprotein affecting iron absorption and metabolism through the binding and degradation of ferroportin, the primary cellular iron exporter. Erythroferrone, a recently identified hormone, inhibits hepcidin transcription during erythropoiesis in animal models. As hepcidin and erythroferrone have not been assessed in human volunteers exposed to HA, the primary objective of the present study was to explore the effects of HA on hepcidin, erythroferrone, andiron availability. Hemoglobin, hepcidin, erythroferrone, and serum iron were measured in unacclimatized healthy youngmen (n = 17, age [mean ± SD] = 23.4 ± 5.6 yrs, body weight = 81.9 ± 13.9 kg) residing at sea level(SL) and HA (19 d, 4300 m). While residing at HA, volunteers consumed energy restricted diets providing 70% of SL weight maintenance energy needs, and engaged in prescribed physical activity to induce a 40% total daily energy deficit. As compared to SL, HA resulted in increased hemoglobin (16.6 ± 0.9 vs. 14.3 ± 0.8 g/dL, P <0.01), diminished hepcidin (4.8± 3.4 vs. 7.5 ± 3.4 ng/mL, P <0.05), and increased erythroferrone (3.1 ± 2.1 vs. 0.9 ± 0.6pg/mL, P < 0.01). Serum iron increased at HA as compared to SL (104.7 ± 34.2 vs. 65.4 ± 17.4μg/mL, P < 0.01). Taken together, these data indicate that prolonged HA exposure in humans results in elevated erythroferrone, which is associated with suppressed hepcidin transcription and increased iron availability.

Support or Funding Information

Research supported by the US Army Medical Research and Materiel Command. The views expressed in this abstract are those of the authors and do not reflect the official policy of the Department of Army, Department of Defense, or the US Government.
Food Group Intake Differs among Adult Food Secure, Low Food Secure and Very Low Food Secure Rural, Midwestern Emergency Food Pantry Users
Authors: Wright Breanne N. , McCormack Lacey A. , Stluka Suzanne , Contreras Dawn , Franzen‐Castle Lisa , Henne Becky , Mehrle Donna , Remley Dan , Eicher‐Miller Heather A.
Published: June 2021

Objective

Previous research has documented the high prevalence of food insecurity, or uncertain access to healthy foods, and poor dietary intake in emergency food pantry (EFP) users. However, no studies have determined whether there are differences in dietary intake when stratified by food security status within the EFP population. This study evaluated associations between food security status and the intake of key nutrients and food groups among EFP clients.

Study Design, Setting and Participants

Adult EFP clients (n=445) from a multistate sample of 24 rural, Midwestern EFPs participated at a single time point from September–November 2014.

Outcome Measures and Analysis

Participants completed a characteristics survey, the 18‐item U.S. Household Food Security Survey Module, and up to three National Cancer Institute Automated Self‐Administered 24‐hour dietary recalls (ASA24s). Adult food security status, classified as food secure, low or very low food secure, was the independent variable while mean intake of energy, vitamins, minerals, and food groups were dependent variables. Multiple linear regression models adjusted for energy and other confounders were used to determine associations between food security status and nutrient/food group outcomes.

Results

Low adult food security was significantly positively associated with total energy (kcal) intake (β=245.81 p=0.02) compared with very low food security. Adult food security was significantly positively associated with whole fruit (cup equivalents) intake (β=0.17 p=0.04) compared with low food security.

Conclusions and Implications

Results suggest that food security, low food security, and very low food security status may be specifically associated with the consumption of key food groups and therefore differences in dietary intake by food security range should be considered in the development of future initiatives that target this population.

Support or Funding Information

This project is supported by the Agriculture and Food Research Initiative competitive grant no. 2013‐69004‐20401 of the USDA National Institute of Food and Agriculture.
Gut microbiota dictate metabolic Fate of Curcumin in the colon
Authors: Li Zhengze , Sun Yue , Song Mingyue , Li Fang , Xiao Hang
Published: June 2021

The metabolic fate of dietary bioactive compounds play critical roles in their health effects. Herein, we determined the metabolic fate of curcumin in the gastrointestinal (GI) tract of mice after long‐term dietary administration of curcumin for 6 weeks. Our results demonstrated that in the GI tract curcumin underwent phase I metabolism to yield three metabolites, namely tetrahydro‐curcumin (M1), hexahydro‐curcumin (M2) and octahydro‐curcumin (M3). Curcumin and these phase I metabolites then were subject to phase II metabolism to yield their corresponding conjugated phase II metabolites. The above‐mentioned metabolism of curcumin occurred in the stomach but to a much less extent than that in the small intestine. Only phase II metabolites of curcumin could be found in the small intestine (i.e., conjugated forms of curcumin, M1 and M2), while only phase I metabolites of curcumin could be detected in the cecum and colon (i.e., M1 and M2). This suggested that gut microbiota might be responsible for the deconjugation of phase II metabolites that converted them back to the corresponding phase I metabolites in the cecum and colon. Indeed, anaerobic fermentation of phase II metabolites (isolated from the small intestine of curcumin‐fed mouse) by both mouse and human fecal bacteria deconjugated them to yield phase I metabolites. Moreover, fecal bacteria were able to transform curcumin to yield M1, M2, and M3, and they can further breakdown these four compounds to yield fission products such as ferulic acid. To confirm the role of gut microbiota in biotransformation of curcumin, curcumin‐fed mice were treated with or without dextran sulfate sodium (DSS) in the drinking water (DSS is an agent capable of altering gut microbiota, and we confirmed the altered microbiota by DSS via next generation sequencing of fecal microbiota). Analyses of colonic content by LC/MS demonstrated that in comparison with the control mice, DSS‐treated mice had a completely different curcumin metabolite profile in the colon, i.e., much higher levels of phase II metabolites and much lower levels of fission products of curcumin. Overall, our results demonstrated that composition of gut microbiota had a profound impact on the biotransformation of curcumin in the colon, which might have a significant influence on the health effects of dietary curcumin, especially in the colon.

Support or Funding Information

This study was partially supported by fund from USDA.
Green Tea Extract Protects Against Hepatic NFκB Activation Along The Gut‐Liver Axis in Diet‐Induced Obese Mice With Nonalcoholic Steatohepatitis By Reducing Endotoxin and TLR4/MyD88 Signaling
Authors: Li Jinhui , Chitchumroonchokchai Chureeporn , Kim Joshua B. , Sasaki Geoffrey Y. , Moller Meredith V. , Bouranis John A. , Bruno Richard S.
Published: June 2021

Green tea extract (GTE) reduces NFκB‐mediated inflammation during nonalcoholic steatohepatitis (NASH). We hypothesized that its antiinflammatory activities would be mediated along the gut‐liver axis in a Toll‐like receptor‐4 (TLR4)‐dependent manner. Wild‐type (WT) and loss‐of‐function TLR4‐mutant (TLR4m) mice were fed a high‐fat diet containing 0 or 2% (w/w) GTE for 8 wk before assessing NASH, NFκB‐mediated inflammation, TLR4 adaptor proteins [myeloid differentiation 88 (MyD88) and TIR‐domain‐containing adapter‐inducing interferon‐β (TRIF)], circulating endotoxin, and intestinal tight junction proteins. In WT mice, phosphorylation of the NFκB p65 subunit and NFκB‐dependent gene expression (MCP‐1, iNOS, MPO) were lower in those provided GTE, and were not different from the lowered levels in TLR4m mice regardless of GTE. Compared with WT controls, TLR4m mice had lower TLR4 mRNA expression, and its expression was lowered by GTE in both genotypes. Protein expression of TRIF, which mediates MyD88‐independent TLR4 signaling, was unaffected by genotype and GTE. In contrast, MyD88 expression was lower in mice fed GTE regardless of genotype. Serum endotoxin was not exacerbated in TLR4m mice, but was lowered by GTE in both genotypes. Similarly, duodenal and jejunal claudin‐1 (CLDN‐1) mRNA expression were not affected in TLR4m mice, but GTE increased their expression in both genotypes. Serum endotoxin was inversely correlated with duodenal and jejunal CLDN‐1 (rp= −0.58 to −0.43, p<0.05), and positively correlated with expression levels of TLR4 and phosphorylated‐p65 (rp= 0.46–0.57; p<0.05). Thus, GTE protects against inflammation during NASH, likely by limiting gut‐derived endotoxin translocation and TLR4/MyD88‐dependent NFκB activation.

Support or Funding Information

Supported by USDA‐NIFA.
Plant Polyphenol Extract Regulated Diacylglycerol Acyltransferase and Tristetraprolin Gene Expression in Cultured Mouse Cells
Authors: Cao Heping , Sethumadhavan Kandan , Anderson Richard
Published: June 2021

Bioactive componentsin plant products have been used for the prevention and treatment of various diseases since ancient history. One of the major classes of bioactive compounds is plant polyphenols, which are found in plant seeds, fruits, leaves and bark. Previous studies have shown that cinnamon is beneficial to people with insulin resistance due in part to the insulin‐like activity of cinnamon polyphenol extract (CPE). The objective of this study was to investigate the effects of CPE, cottonseed extract and insulin on the expression of diacylglycerolacyltransferase (DGAT) and tristetraprolin(TTP), two important gene families whose protein products are related to fatbiosynthesis and anti‐inflammatory response in mammalian cells. DGATs areintegral membrane proteins that catalyze the last step of triacylglycerols (TAGs)biosynthesis in eukaryotic organisms. Plants and animals deficient in DGATs accumulate less TAGs and over‐expression of DGATs increases TAGs. DGAT knockoutmice are resistant to diet‐induced obesity. This study used cultured mouse3T3‐L1 adipocytes and RAW264.7 macrophages. The pre‐adipocytes were induced to oil‐droplet‐filled adipocytes and subsequently treated with insulin (0, 10 and 100 nM) and CPE (0, 10 and 100 μg/ml) for 0, 30, 60, 90 and 120 min. RAW macrophages were treated with CPE (100 μg/ml), insulin (100 nM) and lipopolysaccharides (LPS, 0.1 μg/ml) for 0, 30, 60, 120 and 240 min. Total RNAs were extracted from the cells and converted into cDNAs. TaqMan quantitative real‐time PCR was used to evaluate mRNA levels of mouse DGAT1, DGAT2 and TTP genes using RPL32 and UBQLN3 as internal references. Results showed that DGAT mRNA levels were generally decreased and ZTTP mRNA levels were modestly increased by insulin and CPE in adipoctyes. Insulin and CPE effects on DGAT2 and TTP gene expression were modestly increased but LPS increased TTP mRNA levels up to50‐fold in macrophages. The results from cottonseed extracts will be discussed. Our results support the idea that CPE has insulin‐like effects and suggest apotential positive effect of CPE in reducing fat biosynthesis in adipocytes and increasing anti‐inflammatory activities in macrophages. The significance of this study is that CPE could be potentially useful for obesity prevention.

Support or Funding Information

Supported by USDA‐ARS Quality and Utilization of Agricultural Products Research Program 306 through CRIS 6054‐41000‐103‐00D.
Differential effects of isoproterenol and omecamtiv mecarbil on the contractile properties of unloaded myocytes
Authors: Isaac Joslyn Deeann , Buckley Tori Dawn , Campbell Kenneth S.
Published: June 2021

Patients develop heart failure when their ventricles are unable to maintain adequate cardiac output with normal filling pressures. Omecamtiv mecarbil is a novel small molecule that binds to the myosin head and enhances the rate of Pi release. It is currently in clinical trials as a potential therapy for heart failure. We compared the effects of omecamtiv mecarbil and isoproterenol, a well‐known β‐adrenergic agonist, on the contractile properties of single unloaded cardiac myocytes. Cells were isolated by enzymatic digestion from 3 to 6 month old female Sprague‐Dawley rats and electrically stimulated at 0.5 Hz at 32°C. Sarcomere length shortening profiles were measured using high speed video microscopy. Calcium transients were measured by monitoring Fura‐2AM fluorescence. Experiments measured the effects of 0, 0.1, 1.0, and 10.0 μM omecamtiv mecarbil and 0, 0.01, 0.1, and 1.0 μM isoproterenol. Statistical tests were performed using linear mixed models with Tukey post‐hoc criteria. Isoproterenol increased shortening (p<0.001) and accelerated the rate of shortening and relaxation (both effects, p<0.001). In contrast, 10 μM omecamtiv mecarbil reduced shortening (p<0.001) and slowed shortening and subsequent relaxation (both effects, p<0.001). Our working hypothesis is that at high concentrations, omecamtiv mecarbil increases the duty ratio of myosin heads thereby increasing the number of bound cross‐bridges. These cross‐bridges impose a drag on the myofilaments and slow contraction and relaxation.

Support or Funding Information

AHA GRNT2546003, NSF 1538754, NIH TR000117, APS STRIDE, APS UGSRF, HL115473.