bluuebunny/crossref_metadata_2025 huggingface dataset:
Supplemental ketone metabolic therapy slows tumor growth and increases survival time in mice with metastatic cancer
Authors: Poff Angela M , Ari Csilla , Goldhagen Craig , Seyfried Thomas , D'Agostino Dominic
Published: June 2021

Metastasis is responsible for 90% of cancer‐related deaths in the U.S. Failure to test therapies in natural pre‐clinical models of metastasis has prevented the effective translation of treatments from lab to clinic. Metastatic cancer cells are highly glycolytic, requiring a large supply of glucose for energy. Mitochondrial dysfunction prevents cancer from efficiently using ketones for energy. Furthermore, ketones inhibit cancer cell proliferation in vitro. The metabolic deficiencies of cancer can be exploited with a ketogenic diet (KD) which lowers glucose and elevates ketones in the blood, a strategy that slows cancer progression in animals and humans. We hypothesized that elevating blood ketones with supplemental ketone administration (SKA) would inhibit cancer progression and enhance efficacy of the KD. We tested this metabolic therapy in the luciferase‐tagged VM‐M3 mouse model of metastatic cancer which spreads naturally in an immunocompetent host, mimicking the natural metastatic phenotype. Mice were fed either ketogenic or standard diet with supplemental ketones (1,3‐butanediol or ketone ester.) Tumor growth was monitored by in vivo bioluminescent imaging. SKA slowed tumor growth and increased survival time by 34–51% when delivered with standard or ketogenic diet. We propose that SKA could enhance the efficacy of standard care and improve the outcome of patients with advanced metastatic disease.

Grant Funding Source: Office of Naval Research
The ketogenic diet and hyperbaric oxygen therapy work synergistically to slow tumor growth and increase survival time in mice with systemic metastatic cancer
Authors: Poff Angela M , Ari Csilla , Goldhagen Craig , Seyfried Thomas , D'Agostino Dominic
Published: June 2021

Cancer energy metabolism is characterized by an abnormal dependence on glycolysis and fermentation for energy which can be exploited therapeutically by decreasing glucose availability to the tumor. The ketogenic diet (KD) is a high fat, low carbohydrate diet which decreases blood glucose and increases blood ketones and has been reported to slow cancer progression in laboratory and clinical settings. Tumor hypoxia is a consequence of abnormal vasculature and promotes cancer progression and further increases glycolytic‐dependency. Hyperbaric oxygen therapy (HBO2T) increases oxygen saturation within the tumor and can offset the cancer‐promoting effects of hypoxia. Since these therapies target overlapping metabolic defects within cancer cells, we tested the combined efficacy of KD+HBO2T on cancer progression and survival in a murine model of metastatic disease. KD alone significantly slowed tumor growth and increased mean survival time by 34%. Interestingly, while HBO2T alone did not influence outcome, combining the KD with HBO2T resulted in a supra‐additive effect, significantly slowing tumor growth and increasing mean survival time by 80% in mice with systemic metastatic cancer. Our results support the potential use of KD and HBO2T metabolic therapy as adjuvant treatments for patients with advanced metastatic disease.

Grant Funding Source: Office of Naval Research
Genetics of Serum Carotenoid Levels and Obesity Related Traits in Mexican American Children
Authors: Farook Vidya S. , Reddivari Lavanya , Puppala Sobha , Arya Rector , Fowler Sharon P. , Chittoor Geetha , Mohan Birunda , Jenkinson Christopher P. , Lynch Jane L. , DeFronzo Ralph A. , Blangero John , Hale Daniel E. , Duggirala Ravindranath , Vanamala Jairam
Published: June 2021

Serum carotenoid status is a putative biomarker for fruit and vegetable consumption. The objective of this study was to determine the genetics of serum carotenoid levels and their correlations with obesity related traits in Mexican American (MA) children. We measured serum α‐ and β‐carotenoid levels in 670 non‐diabetic MA children, aged 6–17 years, using UPLC‐PDA. We determined heritabilities for both carotenoids and assessed genetic correlations between them and obesity‐related traits. Carotenoid values were transformed for these analyses, and adjusted for significant effects of age and sex terms. Carotenoid levels were highly heritable (α‐carotene: h2=0.81, P = 6.7 × 10−11; β‐carotene: h2=0.90, P = 3.5 × 10−15). We found significant (P < 0.05) negative phenotypic correlations between carotenoids and four obesity‐related traits: BMI, waist circumference, fat mass (DXA), and triglycerides (range: α‐carotene = −0.19 to −0.12 and β‐carotene = −0.24 to −0.13), and positive correlations with HDL‐cholesterol (α‐carotene = 0.17 and β‐carotene = 0.24). However, all genetic correlations were significantly negative for β‐carotene and these four obesity related traits (range: −0.30 to −0.23), but positive for HDL‐cholesterol (0.31). These results suggest that common genetic factors influence β‐carotene and obesity‐related traits in MA children.

Grant Funding Source: This study was supported by grants from the National Institute of Health (R01 HD049051, HD041111, DK53889, DK42273, P01 HL45522, DK47482, MH59490 and M01‐RR‐01346), Voelcker Foundation and National Research Initiative Grant 2009–55200‐05197 from the USDA National Institute for Food and Agriculture (2009–2012).
Piperidine and pyridine alkaloid actions at fetal muscle‐type nicotinic acetylcholine receptors
Authors: Green Benedict T. , Welch Kevin D. , Lee Stephen T. , Pfister James A. , Panter Kip E.
Published: June 2021

Preventing the normal movement of a developing fetus can result in congenital contracture‐type defects such as those seen in crooked calf syndrome. The desensitization of fetal muscle‐type nicotinic acetylcholine receptors (nAChR) by piperidine and pyridine alkaloids in a developing fetus is one putative mechanism behind the inhibition of fetal movement in animals. In this study, we tested the piperidine alkaloids anabasine and lobeline as well as the pyridine alkaloid myosmine for agonist actions and sensitivity to alpha conotoxins EI and GI antagonism at fetal muscle‐type nAChR expressed by TE‐671 cells using a membrane potential sensing dye based system. The alkaloids acted as agonists in the cell culture experiments. Anabasine and myosmine were sensitive to alpha conotoxin GI, and conotoxins did not block the actions of lobeline. We also tested if these alkaloids can decrease fetal movement in 40 day pregnant goats, dosed IV. Anabasine at 0.8 mg/kg, lobeline at 4 mg/kg and myosmine at 5 mg/kg significantly reduced fetal movement after IV dosing. These results suggest that anabasine and myosmine are nAChR agonists and that the three alkaloids have the potential to reduce fetal movement in animals. This research was supported by USDA/ARS.
Identifying the α7‐nicotinic acetylcholine receptor interactome from a human neuroblastoma cell line
Authors: Mulcahy Matthew John , Barrantes Francisco , Hawrot Edward
Published: June 2021

The α7‐nicotinic acetylcholine receptor (α7‐nAChR) is a ligand‐gated ion channel widely expressed in the mammalian CNS that is associated with numerous physiological functions. α7‐nAChRs are also the principal high‐affinity α‐bungarotoxin (bgtx) binding proteins in the mammalian brain. There is growing evidence that α7‐nAChRs are involved in a number of protein‐protein signaling cascades. The SH‐EP1‐hα7‐Ric‐3 cell line (Valles et al., 2009) is derived from the SH‐EP1 human neuroblastoma cell line which has been transfected with human α7‐nAChR (hα7) and human Resistance to Inhibitors of Cholinesterase 3 (Ric‐3). Bgtx‐sensitive protein complexes were isolated from SH‐EP1‐hα7‐Ric‐3 cells using bgtx affinity immobilization. Total bound protein was eluted, reduced and alkylated prior to digestion with trypsin in‐solution. The resulting peptides were analyzed with an LTQ‐Orbitrap mass spectrometer and data analyzed using ProteoIQ™. More than 20 proteins (1% peptide FDR; 90–100% probability) that were not present in SH‐EP1 controls were identified from multiple preparations of SH‐EP1‐hα7‐Ric‐3 cells. These proteins, which may be involved in receptor function and biogenesis, will serve as a foundation for further investigations of SH‐EP1‐hα7‐Ric‐3 and other human‐derived cells lines as well as primary human tissue. This research is supported by NIH 1R21AG038774, 1S10RR027027, and NSF EPS‐1004057.
Behavioral effects of oxycodone following traumatic brain injury in rats
Authors: Nicholson Katherine L , Greenwood Maria A , Day Nicole L. , Lockhart Roxanne M , Shelton Keith L. , Floyd Candace L.
Published: June 2021

Epidemiological data indicate that drug abuse rates increase following traumatic brain injury (TBI), but the underlying reasons remain unclear. There is overlap in reward pathways and regions commonly damaged in TBI suggesting that TBI could alter risk for drug abuse. Adult male Sprague Dawley rats underwent sham (control) or lateral fluid percussion injury of moderate severity followed by assesment of tolerance development and intravenous self ‐administration behavior. The antinociceptive effects of oxycodone were evaluated using hot plate‐induced paw withdrawal and warm water tail withdrawal models both acutely and following chronic administration of oxycodone. The reinforcing effects of oxycodone were examined using acquisition of oxycodone self‐administration behavior. No consistent differences have been detected in the antinociceptive effects of oxycodone between TBI subjects and sham injured subjects. However the data suggest that the high (0.03 mg/kg/infusion) and intermediate (0.01 mg/kg/infusion) doses of oxycodone resulted in a faster rate and a higher total percentage of TBI subjects acquiring self‐administration. Additionally, the TBI subjects appeared to be less sensitive to oxycodone's effects, self‐administering the highest number of infusions at the 0.03 mg/kg/infusion dose as compared to sham injured subjects. Research supported by DoD grant W81XWH‐11–1‐0374.
Effects of Perfluorocarbon emulsions on microvascular blood flow and O2 transport in a model of severe arterial gas embolism
Authors: Torres Luciana , Spiess Bruce , Filho Ivo Torres
Published: June 2021

Perfluorocarbon emulsions (PFC) have been advocated as a potential treatment of arterial gas embolism (AGE). We hypothesized that PFC can enhance blood flow and oxygenation in massive AGE, due to the surfactant properties of fluorocarbons. We studied systemic and microvascular parameters in cremaster muscle of anesthetized rats via intravital microscopy. Microhemodynamics as well as microvascular and tissue oxygenation were determined before and after treatment with two commercial PFC (Oxycyte, Oxygen Biotherapeutics Inc.; and PHER‐O2, Sanguine Corporation) compared with saline. Blood pressure and heart rate were similar among the groups. Systemic PO2, O2 content and glucose were higher in PFC than in saline group while hematocrit significantly dropped after treatment but was not different among the groups. Arteriolar blood flow went up 58% and 45% with PFC vs. 12% with saline after treatment. In both PFC groups, arteriolar and tissue PO2 were significantly higher compared to control as well as the arteriolar O2 content and O2 delivery. In conclusion, administration of PFC in severe AGE is superior than saline primarily attributable to the surfactant properties which increased considerably air bubble reabsorption. As an additional beneficial effect, arteriolar blood flow was restored, reducing diffusional resistance of O2 in the plasma to yield more O2 to tissues.

Supported by Office of Naval Research.
Effects of resuscitation fluids on endothelial glycocalyx (EG) and coagulation in severe hemorrhagic shock (HS)
Authors: Torres Luciana , Sondeen Jill , Ji Lisa , Dubick Michael , Filho Ivo Torres
Published: June 2021

To study the effects of resuscitation fluids on changes in coagulation and EG in HS, we studied venules of cremaster muscle of anesthetized rats subjected to 1h of HS (40% of blood volume, HEM only) followed by 1h resuscitation with Hextend (HEX), Ringer's lactate (RL), or fresh frozen plasma (FFP). EG thickness was estimated using dextrans of different molecular weights labeled with FITC or Texas Red. Blood gases, hematocrit, cell count, coagulation tests (ROTEM) were performed. EG thickness was 50% lower in HEM, RL and HEX groups. EXTEM: clotting time [CT] was longer for RL compared to other groups. Clot strength [G] in HEM was higher at post‐resuscitation compared to baseline. FIBTEM: maximal clot elasticity [MCE], maximum clot formation [MCF], thrombin generation and G were lower in RL and HEX groups than the others. Resuscitation with FFP was superior in reverting coagulopathy (no differences in CT, G and MCE compared to Sham) and restoring EG thickness. Resuscitation with RL and HEX reestablished mean arterial pressure and tissue perfusion but caused EG shedding and altered coagulation by reducing clot strength and elasticity, and diluting coagulation factors, in contrast to FFP. We showed that ROTEM can indentify changes in coagulation function in massive blood loss and HS‐associated coagulopathy in a rat model that also displayed microvascular changes.

Supported by US Army Medical Research & Materiel Command.
Effects of Resuscitation Fluids following Hemorrhagic Shock on Venular Glycocalyx in Rats
Authors: Filho Ivo Torres , Torres Luciana , Sondeen Jill , Ji Lisa , Dubick Michael
Published: June 2021

The protection mechanisms of resuscitative agents after hemorrhagic shock (HS) are still unclear. For instance, fresh frozen plasma (FFP) may exert its positive effects by replacing lost coagulation factors and by acting upon the endothelium. We compared different resuscitation fluids in their ability to restore the structural integrity of anesthetized rat cremaster venular endothelial glycocalyx (EG) in vivo, as estimated by its thickness using intravital microscopy and fluorescent dextrans. After 1 h of HS (bleeding target: 40% of total blood volume), rats were randomized into 4 groups (6–8 rats/group): HEM‐ no resuscitation; LR– lactated Ringer's; HEX–Hextend and FFP. Baseline EG averaged 0.425 ± 0.015 μm (n=121). After resuscitation, EG was reduced by nearly 70% in HEM, LR and HEX groups to an average 0.132 ± 0.010 μm. However, in rats that received FFP, EG recovered to a thickness of 0.532 ± 0.060 μm, not significantly different from baseline. Syndecan‐1 plasma levels also rose significantly in all groups except in rats that received FFP, where levels returned to baseline. All groups showed significant reductions in venular blood flow, but only HEX rats showed full recovery of flow. The data suggest that plasma but neither colloid nor crystalloid resuscitation support vascular stabilization by reconstitution of EG after HS‐mediated degradation.

Supported by US Army Medical Research & Materiel Command.
NADPH oxidase‐4 mediated ROS generation modulates C2C12 differentiation via ERK1/2 pathway
Authors: Acharya Suresh , Peters Alexander M. , Norton Ann S. , Murdoch Gordon K. , Hill Rodney A.
Published: June 2021

Background

NADPH oxidase 4 (Nox4) enzymatic activity is a major source of intracellular reactive oxygen species (ROS) and plays an important role in balancing oxidation‐reduction within skeletal muscle cells. We hypothesize that changes in Nox4 mediated ROS generation affects the proliferation and differentiation of skeletal muscle cells. Further, ROS plays an important role in cell signaling pathways and this is dependent on the concentration of endogenous ROS in the cells.

Approach

Using mouse myogenic (C2C12) cells, Nox4 concentrations were manipulated via either plasmid mediated over‐expression or siRNA silencing resulting in increased or decreased endogenous ROS concentrations, respectively. Real time RT‐PCR and Western blot demonstrated effective modulation of Nox4 isoform and endogenous ROS production was measured using flow‐cytometry. We further quantified Nox4 protein expression and myogenin using immunofluorescence microscopy. ERK1/2 phosphorylation was determined by Western blot.

Results

Modulation of Nox4 and endogenous ROS concentrations decreased the expression of myogenic differentiation markers, Myf5, Myogenin, MyoD1 and Pax7. This is associated with a decrease in cell differentiation percentage in the early stages of myogenesis. These changes are associated with altered MAPK pathway activation; specifically phosphorylation of ERK1/2. In both over‐expression and silencing of Nox4, we observed significant decreases in ERK1/2 phosphorylation.

Conclusion

Nox4 activity determines optimum levels of ROS that are important for cellular processes including proliferation and differentiation of C2C12 myoblasts. Changes in myogenic differentiation pathways in response to Nox4 modulation are mediated via MAPK activation.

Funding: USDA/NIFA award number 2010–34479‐20715 “Increasing Shelf Life of Agricultural Commodities, ID”
In vitro analysis of the feeding neural circuit in Aplysia after long‐term sensitization training
Authors: Odem Max , Wainwright Marcy , Mozzachiodi Riccardo
Published: June 2021

The effects of long‐term sensitization (LTS) training on defensive responses in Aplysia are well understood behaviorally and cellularly. Conversely, LTS training's effects on feeding are only marginally understood. 24 h after LTS training, the tail‐siphon withdrawal reflex is enhanced and, concomitantly, feeding is suppressed. The goal of this study was to characterize LTS training's effects on the feeding central pattern generator (CPG). During biting, the nerves controlling radula protraction, closure, and retraction express patterns of buccal motor activity (BMPs) generated by the CPG. LTS training's effects on BMP production were examined by recording extracellularly from the I2 (protraction), Rn.1 (closure), and Bn.2,1 (retraction) nerves while activating the CPG via electrical stimulation of the Bn.2,3 nerve. Classification of BMPs as ingestive or egestive has been previously established both in vivo and in vitro. Results indicated no change in total CPG output between trained and untrained animals. However, differences in the types of BMPs generated emerged. Trained animals showed a suppression of ingestive BMPs and a reduced ability of the CPG to generate complete BMPs. These changes in BMP production represent correlates for LTS training‐induced suppression of feeding at the circuit level, and might explain for the changes in biting behavior measured in vivo. Funded by NSF IOS – 1120304.
Membrane trafficking of aquaglyceroporin HC‐3 in erythrocytes from the freeze tolerant anuran, Cope's gray treefrog, Hyla chrysoscelis
Authors: Wetzel Elizabeth Claire , Nickell Philip , Mutyam Venkateshwar , Frisbie James , Goldstein David L , Krane Carissa M
Published: June 2021

Cope's gray treefrog, Hyla chrysoscelis is a freeze‐tolerant anuran which accumulates and distributes glycerol as a cryoprotectant before freezing. We hypothesize that HC‐3, an ortholog of mammalian aquaporin 3, functions as an aquaglyceroporin to facilitate transmembrane flux of water and glycerol. HC‐3 protein is in higher abundance and is preferentially localized to the plasma membrane in RBCs from cold‐acclimated treefrogs as compared to warm‐acclimated animals. The objective of this study is to determine the signal for HC‐3 membrane translocation that occurs during the cold‐acclimation period. After 48 hrs, cultured erythrocytes from H. chrysoscelis were exposed to cAMP (1 uM; 30 minutes), vasopressin (10 IU; 30 minutes), or epinephrine (1 uM; 60 minutes), isolated from the culture media, and fixed on microscope slides. Fluorescent immunocytochemistry showed enhanced HC‐3 membrane localization in cells exposed to epinephrine and cAMP as compared to controls. These data indicate HC‐3 translocation and membrane localization is enhanced in a cAMP‐dependent pathway. This research was supported by NSF Research Grant IOS‐1121457 and the UD University Honors Program.
Permeability to glycerol differs in erythrocytes from freeze‐tolerant Hyla chrysoscelis and freeze‐intolerant Lithobates catesbeianus
Authors: Honer Naava , Finatti Luiz , Frisbie James , Goldstein David
Published: June 2021

Hyla chrysoscelis is a freeze tolerant treefrog that accumulates cryoprotective glycerol during cold acclimation in advance of freezing. In contrast, the bullfrog Lithobates catesbeianus is freeze intolerant and does not accumulate glycerol. We hypothesized that freeze tolerance in Hyla should entail higher cellular permeability to water and glycerol, and that this is facilitated by aqua/glyceroporins. We tested this by assessing permeability, as change in shape, of RBCs from Hyla and Lithobates exposed to water, glycerol, or urea gradients. Timelapse images of RBCs were recorded using MetaMorph software on a Nikon inverted phase contrast microscope, and cell morphology was measured using ImageJ. Initial analyses suggest similar water and urea permeability between species. Both Hyla and Lithobates RBCs react to these stresses with rapid cell shape change from elliptical to spherical, a response blocked by pre‐incubation with 0.3 mM HgCl2. Both species also express a protein reactive to antibody against the Hyla glyceroporin HC‐3, which confers significant glycerol permeability. However, whereas Hyla RBCs become spherical upon incubation in glycerol solution, those from Lithobates do not. The results suggest lower permeability to glycerol and/or different characteristics of the cellular cytoskeleton in Lithobates. Supported by NSF IOS‐1121457 to D. Goldstein and C. M. Krane.
Sensing acid/base conditions via soluble adenylyl cyclase in aquatic animals
Authors: Tresguerres Martin , Barron Megan E. , Barott Katie L. , Ho Jason , Roa Jinae N.
Published: June 2021

Soluble adenylyl cyclase (sAC) is a recently discovered enzyme that is stimulated by bicarbonate ions to produce cAMP. In mammals, sAC is sensor of acid/base (A/B) conditions that modulates diverse physiological process, including but not limited to epithelial ion and fluid transport, sperm flagellar movement, and gene transcription. sAC also acts as an A/B sensor in shark gills, boney fish intestine, and sea urchin sperm. Here we show evidence supporting that sAC plays a similar sensing role in other aquatic organisms. (1) based on genomic and RT‐PCR results, sAC is present in Acropora reef‐building corals. In addition, coral tissue homogenates demonstrate an unusually high capacity to produce cAMP, which is stimulated by bicarbonate and sensitive to the sAC inhibitor KH7. 2) A sAC gene is present in the genome of Pacific oyster Crassostrea gigas, and sAC mRNA is expressed in gill, mantle and hemocytes. 3) sAC protein is present in several shark tissues (e.g. gill, rectal gland, eye). At least in gill and rectal gland, sAC seems to be located both in the cytoplasm and in the nucleus. 4) sAC orthologs are identifiable in EST, TSA, wgs and HTGS databases from diverse aquatic animals such as placozoan, sponge, anemone, acorn worm, sea squirt, chimera fish, skate, coelacanth, gar, catfish, trout and salmon. We propose sAC is an evolutionarily conserved A/B sensor with diverse and important physiological functions in aquatic animals. Funded by NSF grant 122041 to MT, NSF GRFP to MEB, NSF OCE PDF to MLB, and San Diego Fellowship to JNR.
HU decreases diffusible supercoiling density in bacterial chromosome
Authors: Agbleke Andrews Akwasi , Rovinskiy Nikolay , Chesnokova Olga , Higgins Patrick N
Published: June 2021

HU is an abundant protein in bacterial chromosome. It binds DNA and constrains negative supercoiling as well. In E.coli and Salmonella, it exists as a heterodimer composed of alpha and beta subunits. To study in vivo supercoiling that exists in hupA, hupB and hupAhupB double mutants, we employed the gamma delta resolution assay. Supercoil sensors were placed eight sites around the chromosome and resolution assay done to determine diffusible supercoil densities at these sites. Our studies showed that in addition to constraining supercoiling HU also enhances diffusible supercoiling either by its direct interaction with the chromosome or through gyrase activity. Supercoil density was measured to be − 0.035±0.002 in WT then reduced to −0.027±0.006, −0.022±0.007 and 0.013±0.004 in hupA, hupB and HupAhupB double mutants respectively. We observed significant differences between hupA and hupB subunits as well with hupA dimers constrains supercoiling better than hupB. We then measured transcription elongation rates in these mutants using beta galactosidase assay. HU double mutants have reduced transcription elongation rates 26 nuc/sec from WT 38–70 nuc/sec. Though hupA and hupB have similar reduced elongation rates (37 – 46 nuc/sec) alpha‐dimers had a slightly higher rate than beta‐dimers. Taken together, we were surprised that unlike eukaryotes where histone loss activates transcription, in bacteria this seems the opposite.

This work was supported by National Science Foundation
Transcriptional regulator MftR from Burkholderia thailandensis participates in oxidative stress responses
Authors: Gupta Ashish , Grove Anne
Published: June 2021

Burkholderia thailandensis encodes the major facilitator transport regulator (MftR), which is a member of the multiple antibiotic resistance regulator (MarR) protein family. The genomic locus predicts that MftR regulates expression of genes encoding MftR and major facilitator transport protein (MFTP). MFTP is a drug efflux pump, which belongs to the major facilitator superfamily. It is predicted to extrude phenolic compounds and detergents, which can be harmful to this bacterium. Sequence conservation and biochemical analysis has shown that MftR binds the ligand urate, and that urate attenuates DNA binding. Molecular docking was used to identify the ligand binding site, predicting that W11 and R63 are involved in the ligand binding. These residues were also present in the ligand binding site of a homologous transcription factor encoded by Deinococcus radiodurans. By quantitative RT‐PCR it was observed that MftR represses its own expression and that of MFTP, and that this repression is relieved in the presence of urate. MftR also regulates expression of other transcriptional regulators annotated as LysR and SoxR homologs. Since urate is generated under conditions of oxidative stress, and since SoxR is predicted to function in oxidative stress responses, our data suggest a role of MftR in such events.

Source of research support: funded by National Science Foundation
Identify PCBP1‐interacting protein and its novel functional role on human MOR gene expression
Authors: Nahar‐Gohad Pranjal , Sultan Hamidah , Esteban Ysabella , Stabile Alyda , Ko Jane
Published: June 2021

PCBP1 is an expressional regulator of the mu‐opioid receptor (MOR) gene. We hypothesized the existence of a PCBP1 co‐regulator modifying human MOR gene expression by protein‐protein interaction with PCBP1. A human brain cDNA library was screed using the two‐hybrid system with PCBP1 as the bait. RACK1 protein, containing WD domains, was identified. PCBP1‐RACK1 interaction was confirmed via in vivo validation, and by co‐immunoprecipitation with anti‐PCBP1 antibody using human neuronal NMB cell lysate, endogenously expressing PCBP1 and RACK1. Single and serial WD domain deletion analyses demonstrated that WD7 of RACK1 is the key domain interacting with PCBP1. RACK1 overexpression resulted in a dose‐dependent decrease of MOR promoter activity using p357 plasmid containing human MOR promoter and luciferase reporter gene. Knock‐down analysis showed that RACK1 siRNA decreased the endogenous RACK1 mRNA level in NMB, and elevated MOR mRNA level as indicated by RT‐PCR. Likewise, a decrease of RACK1 resulted in an increase of MOR proteins, verified by binding assay. Collectively, this study reports a novel role of RACK1, physically interacting with PCBP1 and participating in the regulation of human MOR gene expression in NMB cells.

*This research was supported by NIH research grant, NSF MRI grant and SHU Biological Research.
Feedback inhibition of 1‐deoxy‐D‐xylulose 5‐phosphate synthase (DXS) regulates the 2‐C‐methyl‐D‐erythritol 4‐ phosphate (MEP) pathway
Authors: Banerjee Aparajita , Wu Yan , Banerjee Rahul , Li Yue , Yan Honggao , Sharkey Thomas D.
Published: June 2021

The 2‐C‐methyl‐d‐erythritol 4‐phosphate (MEP) pathway constitutes an important metabolic pathway for the biosynthesis of isopentenyl diphosphate (IDP) and dimethylallyl diphosphate (DMADP), the precursors for isoprene and other higher isoprenoids. The goal of this study was to understand the regulation of the MEP pathway. It was suggested from previous studies that the first enzyme of the MEP pathway, 1‐deoxy‐d‐xylulose 5‐phosphate synthase (DXS), could have significant regulatory role. In order to test that the DXS enzyme was cloned from Populus trichocarpa and the recombinant protein (PtDXS) was purified from E. coli. An LC‐MS/MS based assay was developed for studying the regulation of PtDXS activity. PtDXS was found to be inhibited by IDP and DMADP. Further studies show that both of these metabolites compete with thiamine diphosphate for binding with the enzyme. Computational analysis shows that IDP and DMADP bind with the enzyme in a manner very similar to the binding of thiamine diphosphate. The feedback inhibition of PtDXS by IDP and DMADP constitutes an important mechanism of metabolic regulation of the MEP pathway. This work was supported by the US National Science Foundation (Grant IOS‐0950574). Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation.
Redox Control of N‐linked Protein Glycosylation
Authors: Petkovich Daniel Adam , Fomenko Dmitri
Published: June 2021

The Endoplasmic Reticulum (ER) is the cellular compartment where proteins are folded and glycosylated. This compartment is characterized by very intense and diverged redox processes that are catalyzed by numerous thiol oxidoreductases. Asparagine‐linked glycosylation is an essential redox‐regulated process of posttranslational protein modification within the ER, catalyzed by the multiprotein complex known as oligosaccharyltransferase (OST). The functions of most of the OST subunits are poorly understood. OST3/6‐like proteins are thioredoxin fold ER membrane‐linked thiol oxidoreductases involved in redox control of N‐linked protein glycosylation as components of OST complex. The exact biological function of these subunits is not well defined. The purpose of this research is to demonstrate the biological roles of OST3/6 – like proteins, clarify their contribution in metal metabolism and determine the impact of OST3/6 proteins deficiency in ER stress development. OST3/6 mutants show affected growth rate, cell wall integrity and presence of ER stress. We demonstrated that the OST3/6 double mutant is not viable, but OST3/6 mammalian homologs, IAP and N33, and reductases (TRXs) or isomerases (PDIs) can complement OST 3/6 mutation. Thus, OST3/6 –like proteins can serve as strong reductases or weak isomerases. This work was supported by NSF grant DBI‐ 1156692 and the Redox Biology Center REU program.
Home food gardening, fruit and vegetable intake, and weight status in pre‐kindergarten children
Authors: Martin Roanna Gail , Partington Susan N , Lacombe Donald , Waterland Nicole
Published: June 2021

Lack of access to fruits and vegetables in the home and low intake of fruits and vegetables have been associated with poor dietary quality and obesity in children. The objective of this study was to determine the associations among existence of a home food garden, intake of fruits and vegetables, and weight status in young children.

Anaylses included survey data (food gardening questions from the 2009 National Gardening Survey, the Block Kids FFQ, family socio‐demographic characteristics), and height and weight for 124 pre‐kindergarten children, participants in the West Virginia Early Childhood Obesity Prevention Project. Comparisons of group means (t‐test, p≤ 0.05) were used to examine differences in fruit and vegetable intake and associated micronutrients and weight status of children between families with and without a home garden.

Children in families with a home garden consumed significantly more cups of dark green vegetables (0.20 ± 0.03 vs 0.11 ± 0.01) and deep yellow and orange vegetables (0.10 ± 0.01 vs. 0.06 ± 0.01) per day. Associated micronutrients; vitamin K, beta‐ and, alpha‐carotene were also significantly greater. Mean weight status (CDC 2000 BMI percentiles for age and gender) was not significantly different. Home food gardening has the potential to positively impact the diet quality of young children. This project was supported by USDA AFRI grant #2011–68001‐30049.
Does mere exposure change implicit associations between whole grains and taste?
Authors: De Leon Angela , Burnett Dustin J. , Horn William , Keim Nancy
Published: June 2021

The health benefits of whole grain (WG) consumption are well documented. Current Dietary Guidelines recommend that individuals consume ≥ 3 servings per day. Despite increased variety, availability, and promotion of WG products, less than 5% of Americans consume the recommended amounts. Healthy adults were provided weekly market baskets of a variety of WG products in recommended amounts for home use. We hypothesized that exposure to WG foods would change implicit associations between WG and taste. Intake was measured by calculating disappearance and verified by a daily log. Administered at baseline and at the end of the 6‐week intervention, the Implicit Association Test (IAT) is a computerized paradigm that measures indirectly the strength of association between pairs of concepts: (a) two contrasted target categories (WG and refined grain (RG) food images) and (b) two contrasted attribute categories (pleasant words and unpleasant words relating to taste) via a classification task. Participants rapidly classify stimuli representing target and attribute into categories. Response time (milliseconds) was used to calculate IAT D scores. Paired t‐tests showed that average D scores at the end of the intervention showed a significant positive shift in implicit attitude towards WG (p≤0.05) in those who initially associated WG with negative taste. This suggests that mere exposure to WG products over an extended period of time in a free living situation can improve automatic attitudes toward WG and may result in increased consumption. Supported by USDA 5306–51530‐019–00D.
Iron‐dependent regulation of mitochondrial form and function
Authors: Jeevananthan Athavi , Rensvold Jarred W. , Ong Shao‐En , Carr Steven A. , Mootha Vamsi K. , Pagliarini David J.
Published: June 2021

Mitochondria play vital roles in cellular metabolism and signaling, and their composition and function must change in response to environmental conditions and nutrient demands. However, the signals that initiate this mitochondrial adaptation remain largely unclear. Our matched genomic and proteomic analyses reveal that proteins involved in iron homeostasis are highly coordinated with mitochondrial biogenesis and that iron deprivation alone results in a systematic loss of mitochondrial transcripts and proteins. Moreover, we determined that the reintroduction of iron to iron‐deprived cells resulted in the complete recovery of mitochondrial transcripts, proteins and respiratory function. This indicates that our observed mitochondrial response to iron deprivation is a calibrated adaptive response and not merely irreversible cellular damage. Taken together, our results reveal that iron deprivation drives a widespread, adaptive transformation of mitochondrial form and function. This research was supported by a Searle Scholars Award, and grants from the NIH and USDA.
Human Presence Dramatically Alters the Physiological Response to Diving in the Turtle, Trachemys scripta
Authors: Williams Cassondra L. , Hicks James W.
Published: June 2021

The objective of this study was to understand the physiological response of red‐eared sliders to investigator presence during routine activities. We measured heart rate continuously in red‐eared sliders during undisturbed and disturbed activities. EKG leads were implanted through the carapace above and below the heart and then attached to a self‐contained, small data logger, which was glued to the carapace. After recovery, turtles were released into tanks where they could swim, dive, and bask. Times for all persons entering and exiting the room were recorded. Turtle diving behavior was recorded by a time depth recorder and a video camera continuously recording images. Data recorded by the TDR and data logger were synchronized and analyzed during different activities. Turtles resting on the surface and at the bottom of the tank had very low heart rates, from five to 10 beats per minute (bpm). During activity (swimming and diving), heart rate increased 50 to 60% (8 to 15 bpm). However, when an investigator was present in the room and near the tank, heart rate increased two to threefold above resting (12 to 25 bpm). The presence of investigators in the room with turtles dramatically impacted their heart rate which may have unintended and confounding effects during experiments. This work was supported by a NSF grant IOS 1121324.
Impact of DHA status on serum markers of obesity and inflammation in pregnant women (1001.4)
Authors: Chekal Jessica , Harris Mary , DiRusso Concetta
Published: June 2021

Different dietary fatty acids impact disease prevention and susceptibility. Among these, the N‐3 (omega‐3) long chain polyunsaturated fatty acids are essential nutrients and are proposed to be health promoting while saturated fatty acids are often associated with obesity, metabolic syndrome, insulin resistance and cardiovascular disease. Omega‐3 fatty acids are critical for fetal and newborn neurodevelopment and must be acquired from the mother by placental transfer. The purpose of this research was to examine the effects of omega‐3 fatty acid status in pregnant women on biomarkers of obesity and inflammation. RBC fatty acid composition and a complete dietary assessment was employed to determine DHA status and intake. Total fatty acid analysis was also performed on blinded maternal plasma samples. The levels of specific fatty acid species were then compared with the levels of the targeted biomarkers of obesity and inflammation. We found significant correlations between: palmitate and leptin levels; palmitate and insulin levels; and DHA and lipocalin 2 levels. This research is of value to understand the impact of DHA status in mid‐pregnancy on adipogenic biomarkers.

Grant Funding Source: Supported by: Arnold and Mabel Beckman Foundation, USDA Hatch and NIFA
Longitudinal association of dairy consumption with changes in blood pressure and incident hypertension (1018.3)
Authors: Wang Huifen , Troy Lisa , Fox Caroline , Meigs James , McKeown Nicola , Jacques Paul
Published: June 2021

We examined the longitudinal association between dairy consumption and changes in blood pressure (BP) and incident hypertension (HTN) among Framingham Heart Study Offspring Cohort members who were free of HTN and participated in up to 4 study exams from 1998 to 2008. At each exam, dietary intake was assessed by a validated food frequency questionnaire, BP was measured using standardized procedures, and anti‐hypertensive medication use was obtained by physician‐elicited self‐report. HTN was defined as systolic BP (SBP) 蠅140mmHg, or diastolic BP (DBP) 蠅90mmHg, or anti‐hypertensive medication use. We used repeated‐measure regression and discrete‐time hazard regression to examine the associations of dairy consumption with the annualized BP change (n=2075) and the risk of incident HTN (n=2340). Adjustments included demographic, lifestyle and metabolic factors, and/or medication use, as appropriate. Greater intakes of total dairy, total low fat/fat free dairy, low fat/skim milk, and yogurt were significantly associated with smaller annualized SBP increments; only total low fat/fat free dairy intake was inversely related to changes in DBP. Over an average of 15‐year follow‐up, 1026 participants developed incident HTN. Each serving consumption of yogurt was associated with 6% (1%‐10%) reduced risk of incident HTN. Dairy consumption, as part of a healthy diet pattern, may benefit BP control and HTN prevention.

Grant Funding Source: Supported by NHLBI NO1‐HC‐25195 and USDA agreement 58–1950–0‐014
Is eating breakfast associated with increased fruit and whole grain intake in young women? (1021.11)
Authors: Widaman Adrianne , Souza Elaine , Forester Shavawn , Keim Nancy
Published: June 2021

Two key recommendations from the 2010 Dietary Guidelines (DG) focus on increasing fruit (F) and whole grain (WG) intake. However, only 15% of adult Americans meet the daily WG recommendation of 3 oz equivalents (eq). In addition, 58% of adults consume less than the DG F recommendation. Our study strictly defined breakfast eaters (BE) as consuming 蠅 15% of energy intake between 4‐10am. Breakfast skipping (BS) was defined as no food intake between 4‐10am for 蠅4 d/wk. Dietary intake was measured with 3 multi‐pass, 24 h recalls. A median split was used to separate high from low consumers of WG and F. Independence was tested with Pearson’s Chi‐squared. Consumption of WG oz eq averaged 2.11 ± 0.17 or 1.19 ± 0.25, for BE or BS respectively. Mean total F intake was 1.99 ± 0.23 servings in BE and 0.98 ± 0.16 servings in BS. BS was associated with lower intake of WG (p=0.012) and lower total F consumption (p<0.0005). Regularly eating breakfast is associated with a diet more aligned with the DG. Randomized control trials are needed to determine if a causal relationship between BE and better diet quality exists.

Grant Funding Source: Supported by NIFA grant #2009‐35215‐05364 and USDA intramural funds Project #5306‐51530‐019‐00D
Weekend intake is associated with poorer diet quality, and increased intake energy and nutrients to limit and poorer food choices in adults: National Health and Nutrition Examination Survey 2001‐2008 (1023.4)
Authors: Ledet Taylor , O'Neil Carol , Nicklas Theresa , Fulgoni Victor
Published: June 2021

National Health and Nutrition Examination Survey data (2001–2008) were used to assess the differences between weekend and weekday intake by adults (19y+; N=18,988). Intake was determined using a single 24 hour recall. Diet quality was determined using the Healthy Eating Index‐2005 (HEI). Covariate (DRI age group, gender, ethnicity, poverty index ratio level, physical activity level, and weight status) adjusted regression analyses were conducted to generate beta coefficients. Weekend intake was associated with a lower (p<0.001) HEI score (‐2.2±0.3 [SE]), and lower (p<0.001) intakes total fruit (‐0.1±0.02 cup eq) and milk (‐0.1±0.02 cup eq). Weekends were also associated with higher (p<0.001) intakes of energy (103±20 kcal), total fat (4.7±1.0 g), SFA (1.5±0.4 g), MUFA (2.0±0.4 g), PUFA (0.8±0.2), solid fat (4.3±0.7 g), cholesterol (30.2±4.3 mg), SoFAAS (95.8±11.4 kcals; 2.1±0.3% total energy), alcohol (4.2±0.6 g), and total egg (0.1±0.02). Since data suggest that weekend eating was associated with poorer diet quality and increased consumption of some nutrients to limit, it is important to determine weekend and weekday intake when characterizing the intake of a population.

Grant Funding Source: Supported by LSU College of Agriculture and USDA.
Intake of trans fatty acid isomers found in ruminant fat versus industrial sources differentially impact concentrations of fatty acids in erythrocytes (1025.17)
Authors: Gebauer Sarah , Matthan Nirupa , Lichtenstein Alice , Baer David
Published: June 2021

Evidence suggests concentration of trans fatty acids (TFA) in erythrocyte (RBC) membranes is strongly predictive of coronary heart disease (CHD) risk. Some data suggest a direct association with industrially‐produced TFA (iTFA) and CHD risk, and an inverse or no association with naturally‐occurring TFA in ruminant fat (rTFA). Differential incorporation of TFA isomers in RBC membranes from iTFA and rTFA may account for differences in CHD risk reported between the two sources; however, controlled dietary interventions comparing iTFA and rTFA have not measured RBC TFA isomers. In a double‐blind randomized‐controlled crossover feeding trial healthy adults (N=106; 47M, 59F) were fed 4 diets for 24 days each: 0.1% en mixed isomers TFA (control), 3.0% en highly‐purified vaccenic acid (VA), 3.0% en mixed isomers iTFA, and 1.0% en c9,t11‐CLA. Total fat (34% en) and other macronutrients were matched across diets. RBC VA was highest following the VA diet (P < 0.05), while elaidic acid was highest following the iTFA diet (P < 0.05). RBC c9,t11‐CLA was highest after the CLA diet (P < 0.05), followed by the VA diet (P < 0.05), indicating some conversion of VA to c9,t11‐CLA resulting from the dietary VA. In conclusion, highly‐purified dietary VA and c9,t11‐CLA differentially impact RBC fatty acid profiles compared to iTFA. Differences in RBC fatty acids impact membrane fluidity which may explain differences of rTFA and iTFA on CHD risk.

Grant Funding Source: Supported by: USDA, Nestle, National Dairy Council, Dairy Australia
Diet quality in community dwelling obese women following a behavioral‐based diet, tai chi, and resistance training intervention (1026.7)
Authors: Taetzsch Amy , Lobuono Dara , Quintanilla Dinah , Maris Stephen , Letendre Jonathan , Picard Allison , Mahler Leslie , Xu Furong , Delmonico Matthew , Lofgren Ingrid
Published: June 2021

Up to 48% of older adults may be at increased nutrition risk. This is a concern as the number of older adults is projected to double by 2030 and obesity is suspected to rise to over 20% in the same population. The purpose of this analysis is to examine the impact of a diet and exercise intervention on diet quality in obese women in an urban setting. There were 26 women with mean age of 65.3 ± 8.1 years and BMI of 38.0 ±4.6 kg/m2 who completed a community based diet and exercise intervention. Seventeen women, 65% African‐American, were assigned to the intervention group (EXD) and participated in 12 weeks of Tai Chi (TC), resistance training (RT), and behavioral based diet intervention. Nine women were assigned to the control group (CON). The Dietary Screening Tool (DST) was used to assess diet quality; higher scores indicate better quality. Using an independent t‐test the EXD group had a significantly higher diet quality compared to the CON group (66.5 ± 10.2 vs. 53.5 ± 13.8; p=0.025) at post ‐intervention. Results from this analysis indicate that a behaviorally‐based diet, TC, and RT intervention may result in improved diet quality, however further research is needed.

Grant Funding Source: College of Environment and Life Sciences Community Access to Research and Extension Services, USDA
Effect of the porcine IGF2 intron3 G3072A substitution on development and growth of skeletal muscle (1033.4)
Authors: Clark Daniel , Clark Diana , Beever John , Dilger Anna
Published: June 2021

A previously identified SNP that disrupts a binding site for a repressor, ZBED6, within intron 3 of the maternally imprinted IGF2 gene, (IGF2 G3072A) results in increased muscle but decreased fat accumulation in swine. Our objective was to determine how this mutation alters gene expression and muscle fiber types during prenatal and postnatal growth. Sows (AA) were bred to a heterozygous (AG) boar. For fetal samples, sows were euthanized at 60 and 90 days of gestation to obtain fetuses. Male and female pigs were also euthanized at birth (0d), weaning (21d), and at a market weight (175d). At 175d, muscle weights were increased approximately 5‐10% (P < 0.01) in pigs with paternal A compared to G alleles. As expected, IGF2 expression was increased 1.5 ‐ 4.5 fold (P < 0.05) in pigs possessing paternal A compared to G alleles at 0d, 21d and 175d. Unexpectedly, IGF2 expression tended to have a 1.5 fold increase (P < 0.10) in fetuses with paternal A compared to G alleles at 90 days of gestation. IGF2 was not different (P > 0.10) in fetuses at 60 days of gestation. At all time points, IGF1 expression was not affected (P > 0.10) by IGF2 allele. Myostatin, ZBED6, IGF1R, IGF2R, and Myosin Heavy Chains (1, 2a, 2x, 2b) expression were also not affected (P > 0.1) by IGF2 allele at 175d. These data suggest that increased IGF2, and not increased IGF1, is directly responsible for increased muscle weights observed in market weight animals.

Grant Funding Source: Supported by NIFA