bluuebunny/crossref_metadata_2025_split huggingface dataset:
Purple‐fleshed potatoes suppress colon‐systemic oxidative stress/inflammatory markers via alternations in the gut bacterial signature
Authors: Reddivari Lavanya , Kim Sung Woo , Radhakrishnan Sridhar , Yang Peiying , Knight Robert , Vanamala Jairam 
Published: June 2021

Purple‐fleshed potatoes (PP) are rich in putative anti‐oxidant/antiinflammatory anthocyanins. We hypothesized that PP, even after processing, suppress colon‐systemic oxidative stress/inflammatory markers by altering gut bacterial signature and metabolite profile. To test this hypothesis, 40 pigs on high‐fat diet (HFD) for 12 wk were provided with one of the five diets: HFD, white‐fleshed potato (WP) or PP chips at 10/20% for 5 wk. Serum, distal colon mucosa/digesta and fecal samples were utilized to analyze oxidative stress (MDA; colorimetric assay) and inflammatory (TNFα; ELISA and prostaglandins; LC‐MS/MS) markers, gut bacteria (pyrosequencing) and metabolites (LC‐MS). All diets, except WP (20%), suppressed (p = 0.04) systemic levels of TNFα compared to HFD. However, only PP suppressed MDA and colonic PGE2 levels (p ≤ 0.05). Potato diets increased members of Clostridia and Lachnospiraceae. Only PP increased distal colon bacteroidetes to firmicutes ratio (BFR) compared to HFD. This is particularly important as BFR is reduced during obesity. Indeed, MDA and TNFα elevated during obesity positively correlated with Firmicutes (0.518, p = 0.002; 0.380, p = 0.024, respectively) and MDA negatively correlated with BFR (−0.413, p = 0.036). Preliminary metabolomics data revealed 4350 metabolites in both distal colon and feces and the correlations with gut bacteria are being carried out.Grant Funding Source: National Research Initiative Grant 2009–55200‐05197 from the USDA National Institute for Food and Agriculture (2009–2012)
Relationship between Parental Feeding and Child Eating Styles in Low‐income Families with Preschool Age Children
Authors: Vollmer Rachel L , Mobley Amy R 
Published: June 2021

While parental feeding styles have been associated with children's dietary intake and weight status, the objective of this study was to determine how a child's eating style is related to their parent's feeding style. Parents of preschool children (n=43) completed the Caregiver Feeding Style Questionnaire to determine parental feeding style and the Child Eating Behaviour Questionnaire (CEBQ) to determine their child's eating style. Authoritarian (35%) and indulgent (30%) feeding styles were most common, and the authoritative (14%) feeding style was least common among parents. Children of indulgent parents had higher CEBQ scores for enjoyment of food (r=.453, p= 0.01) and lower scores for slowness in eating subscales (r=‐.430, p=0.01), which have both been linked to higher child BMI. Children of authoritarian parents had higher CEBQ scores for satiety responsiveness (r=.673, p=0.01) and slowness in eating (r=.600, p=0.01), and lower scores for enjoyment of food (r=‐.477, p=0.01), which have been associated with desirable child weight status. Interestingly, children of authoritarian parents scored higher on emotional over eating (r=.522, p=0.01), emotional under eating (r=.343, p=0.05), and food fussiness (r=.481, p=0.01), which may be why these parents use a controlling feeding style. Further research should determine the direction of the relationship between parent feeding and child eating styles.Grant Funding Source: Subcontract to USDA/AFRI Grant 2010–85215‐20662
Acute phytoecdysteroid treatment increases PI3k‐Akt signaling in aged mouse skeletal muscle
Authors: Zwetsloot Kevin A , Lawrence Marcus M , Arthur Susan T , Nieman David C , Lila Mary Ann , Grace Mary , Shanely R. Andrew 
Published: June 2021

Activation of the PI3k‐Akt pathway leading to protein synthesis is diminished in aged skeletal muscle. Phytoecdysteroids, in particular 20‐hydroxyecdysone (20E), from the plant Ajuga turkestanica increases protein synthesis in C2C12 muscle cells. We previously reported that 21 mo old mice fed phytoecdysteroids for 28 d display larger fiber cross‐sectional area in the triceps brachii and plantaris muscles. The objective of this study was to determine the extent to which an acute dose of phytoecdysteroids activates PI3k‐Akt signaling in skeletal muscle of aged mice. Aged male C57BL/6 mice (20 mo) received a single gavage dose (50 mg/kg bw) of A. turkestanica extract (ATE), 20E, or vehicle (CT) and skeletal muscles were harvested 90 min later. Western blotting performed on gastrocnemius muscles revealed a significant 52% increase in Akt phosphorylation in 20E‐treated skeletal muscle of aged mice, compared to CT (p=0.006). The phosphorylation of p70, 4EBP1, and S6 ribosomal protein (downstream targets in the PI3k‐Akt pathway) were unaltered at this time point. In conclusion, these data suggest that a single dose of 20E activates Akt, a central component of the PI3k‐Akt pathway, in sedentary aged skeletal muscle.Supported in part by a USDA Kannapolis Scholars Fellowship.
Physical activity behaviors and cognitions of parents of preschoolers (1021.6)
Authors: Spaccarotella Kim , Martin‐Biggers Jennifer , Hongu Nobuku , Worobey John , Byrd‐Bredbenner Carol 
Published: June 2021

This study examined PA behaviors, cognitions, barriers, and strategies for coping with barriers by 137 parents of preschoolers via survey (n=137) and focus groups (n=23). Parents spent 4.2±2.2 SD days/week engaging in vigorous play with children, with 2/3 spending <3 hours/week in vigorous play. Time spent in active play rose significantly (p<0.01) as parent education increased but did not differ based on primary language (n=84 English, n=50 Spanish), location (n=65 AZ, n=72 NJ), or parent age. Preschoolers spent 2.4±1.7SD hours/day watching TV. Focus group parents reported using TV to occupy children while performing chores, during inclement weather, and before bed. Most believed TV time should be limited but were unsure of alternatives. All felt active play with their children was important for modeling healthy behaviors. Many considered educational TV exempt from TV time limits. Commonly‐cited barriers to increasing time spent in active play with their children included lack of indoor space, lack of safe outdoor play areas, limited time, and health concerns with being outdoors, such as asthma and insect bites. Coping strategies included active video games, school recreation programs, dancing to music, and visiting indoor playgrounds. Results suggest that parents with less education may especially benefit from strategies that help overcome barriers to modeling positive PA behaviors to their young children.Grant Funding Source: Supported by USDA, National Institute of Food and Agriculture, Grant Number 2011‐68001‐30170
Osmotic regulation and tissue localization of the myo‐inositol biosynthesis pathway in tilapia (Oreochromis mossambicus) larvae (1182.6)
Authors: Sacchi Romina , Gardell Alison , Fernando Villarreal , Li Johnathon , Kültz Dietmar 
Published: June 2021

The myo‐inositol biosynthesis (MIB) pathway converts glucose‐6‐phosphate to the compatible osmolyte myo‐inositol (MI), which protects cells from osmotic stress. We exposed tilapia larvae just after yolk sac resorption to various hypersaline environments and recorded robust induction of the enzymes that constitute the MIB pathway, myo‐inositol‐phosphate synthase (MIPS) and inositol monophosphatase 1 (IMPA1). Strong up‐regulation of these enzymes is evident at both mRNA (quantitative real‐time PCR) and protein (densitometric analysis of Western Blots) levels. The highest level of induction of MIB enzymes occurs at the highest salinity that larvae were exposed to (90ppt). Less severe osmotic stress causes proportionally lesser induction of the MIB pathway. Two distinct MIPS mRNA variants are present in tilapia larvae and both are induced at comparable levels for all the salinity challenges (34, 70 and 90ppt). Immunohistochemical localization of IMPA1 protein in sagittal sections of salinity stressed and control larvae identified tissues that are particularly potent in inducing the MIB pathway. These tissues include the skin, gills, eye (retina) and heart. We conclude that the MIB pathway plays an important role in protecting larval stages of tilapia from hyperosmotic stress. Therefore, tilapia larvae represent excellent in vivo models for studying physiological mechanisms by which the MIB pathway is regulated. Supported by NSF grant IOS‐1049780.Grant Funding Source: NSF IOS‐1049780
Optimizing two‐component light‐switchable systems for controlling gene expression in Escherichia coli (991.2)
Authors: Stork Devon , Schmidl Sebastian , Tabor Jeff 
Published: June 2021

Light‐induced gene expression is a powerful tool for manipulating cells because it allows for precise spatial and temporal control of cellular behavior. A light‐sensitive two‐component system based on the Synechocystis PCC6803 phytochrome CcaS and the cognate response regulator CcaR has been previously constructed in Escherichia coli.1 The chromophore‐binding domain of CcaS performs a reversible photoconversion between a green (~535 nm) and a red (~672 nm) light absorbing form. The green form phosphorylates CcaR, which then induces expression of the cpcG2 promoter by binding to a conserved direct repeat designated the G‐box. Green light‐induced transcription from PcpcG2 was about ~4.3‐fold for this original construct.In order to engineer a constitutively expressed green/red light‐switchable two‐component system with improved fold‐inducibility, we built different ccaS expression cassettes using the constitutive promoter J23106 and RBSs with various translation initiation rates. This library was first screened against an aTc‐inducible expression system for CcaR resulting in a ~40 fold‐inducibility. After screening different ccaR expression cassettes using the constitutive promoter J23100 and RBSs with various translation initiation rates, a fold induction as high as ~80‐fold for a specific CcaS/CcaR‐expression pair could be observed.There are over 10,000 two‐component systems in nature, and discovering the design and expression rules for these systems will allow engineering of a wide variety of genetic circuits for practical applications. The increase in fold‐induction in this system makes light‐induction a more powerful method of spatiotemporal control of living cells and aids precise cellular studies in synthetic and systems biology.Grant Funding Source: Supported by an NSF award from the Division of Biological Infrastructure No. 1004853.
Glial Cell Line Derived Neurotrophic Factor Protein Secretion by Skeletal Muscles is Altered by Activity of L‐Type Calcium Channels
Authors: Boynton Alicia M. , Spitsbergen John M. 
Published: June 2021

Glial cell line‐derived neurotrophic factor (GDNF) is an important signaling molecule throughout the peripheral nervous system. GDNF protein is produced and secreted by skeletal muscle cells and helps maintain motor neuron innervation at the neuromuscular junction. Treatment with exogenous GDNF prevents denervation which occurs with aging and neurodegenerative disease. The therapeutic potential of GDNF cannot be fully explored without understanding the mechanisms by which GDNF protein production is regulated. Although the specific cellular events necessary for regulation of GDNF secretion remains to be elucidated, previous studies from our laboratory have shown that increased skeletal muscle activity significantly alters GDNF protein levels in vivo and in vitro. The primary objective of this study was to determine the role of calcium in regulating GDNF protein secretion by skeletal muscle cells. Skeletal muscle cells (C2C12) were grown in culture and allowed to differentiate into myotubes which display a contractile phenotype. The following treatments were given, Nifedipine to block the voltage‐gated L‐type calcium channels, and Bay K8644 to enhance calcium influx through L‐type calcium channels. Both Western Blot and ELISA were employed for protein identification and GDNF protein quantification respectively. Preliminary results indicate that treatment of myotubes with Bay K8644 (100μM) caused an increase in GDNF secretion to 170% of that in untreated controls. Treatment of cells with Nifedipine blocked the stimulatory effect of Bay K8644, and treatment with Nifedipine alone (10μM and 100μM) inhibited GDNF secretion to 77% of that in control cells. The results suggest that secretion of GDNF protein by C2C12 myotubes is regulated by intracellular calcium levels. Acquiring a greater understanding of the role that calcium plays in regulating GDNF production may help to identify potential sites for therapeutic intervention to increase or decrease GDNF production.Support or Funding InformationNSF Grant DBI‐1062883 and NIH Grant 1R15AG022908‐01A2
Crystal structures of acetylated HRas K104 mimic K104Q and mutant K104A suggest unique role of K104 in interlobe communication across HRas
Authors: Marcus Kendra , Johnson Christian , Sanchez Jaquelyn , Mattos Carla 
Published: June 2021

Ras GTPases belong to a subfamily of signal transduction switches found in the Ras superfamily. Three isoforms of Ras (H‐, K‐, and N‐Ras) are expressed to regulate cellular proliferation, growth, and apoptosis. Unregulated activity of these proteins is closely related to disease, particularly the growth of tumors.1 The GTP/GDP cycle is a common regulatory feature of signaling output across GTPases. Exchange of GDP for GTP within the Ras active site results in an active signaling cascade. GAPs accelerate the hydrolysis of GTP to GDP, turning off the signal. In addition to exchange factors and activating proteins, the Ras GTPase cycle is also controlled by post‐translational modifications. An acetylation mimic (K104Q) in KRas G12V has been shown to negate oncogenic growth of the double mutant cell lines. However, KRas K104A/G12V does not reduce the oncogenicity of the cell line.2 The purpose of this study is to investigate the structural‐functional implications of K104Q and K104A mutations in HRas through X‐ray crystallography and to obtain their intrinsic hydrolysis rates through kinetic experiments with γ‐32P‐GTP. These data show a novel hydrogen‐bonding network present in K104Q, but not WT or K104A HRas. Intrinsic hydrolysis of GTP to GDP is significantly impaired in HRas K104Q, but is not affected by a K104A mutation. The unique network may control helix 5 motility in HRas, resulting in a lower intrinsic hydrolysis rate. Connectivity from K104Q towards helix 5 passes through a region of amino acids that are different between the H‐, K‐, and N‐ Ras isoforms. This suggests that Ras acetylation of K104 may affect the function of the isoforms in subtly different ways.Support or Funding InformationThis work is funded by a grant from the NSF MCB‐1244203
Defining Thermophilic Glucan Dikinases in Cyanidioschyzon merolae
Authors: Brizzee Corey O. , Husodo Satrio , Gentry Matthew S. 
Published: June 2021

ObjectiveThe goal of this project is to elucidate the mechanism of action of starch dikinases in carbohydrate metabolism using the red alga Cyanidioschyzon merolae (Cm) as a model system.SignificanceHighly regulated reversible starch phosphorylation is key for starch metabolism in photosynthetic organisms such as plants and algae. Defining the enzymes that are involved in reversible starch phosphorylation in C. merolae will allow us to engineer strategies to harness starch production in a more efficient manner for many industrial applications.MethodsBioinformatics to identify conserved functional domains with orthologs from different kingdoms. Radiometric kinase assays to measure specificity and kinetic parameters of starch dikinases. Differential Scanning Fluorimetry (DSF) to characterize glucan binding and protein thermal stability. Cell biology techniques such as gene suppression, synchronized cultures, mRNA extraction, and immunofluorescence. Structural analysis through Small Angle X‐Ray Scattering (SAXS), protein purification, and western analysis.ResultsWe show here that C. merolae GWD shares conserved functional domains with orthologs from many different kingdoms. Validating Cm‐GWD is a functional glucan dikinase, results show robust Cm‐GWD specificity towards phosphorylating the C6 position of glucose residues. Radiometric‐ATP incorporation assays reveals that recombinant Cm‐GWD is as efficient as Arabidopsis thaliana (At)‐GWD at phosphorylating starch. Lastly, we utilize SAXS to structurally characterize each domain of the multi‐domain Cm‐GWD.ConclusionWe have defined C. merolae growth and optimal starch accumulation parameters as well as cell localization of enzymes involved in reversible starch phosphorylation. Furthermore, we have characterized gene expression of Cm‐GWD via mRNA levels during synchronous cultures. Additionally, this study exposes biochemical targets to enhance starch metabolism in planta. The biochemical properties of Cm‐GWD enable increased starch metabolism not only in C. merolae but in exogenous plants.Support or Funding InformationKSEF grants KSEF‐2268RDE‐014 and KSEF‐2971‐RDE‐017; NSF Grants IIA‐1355438 and MCB‐1252345
Perceived Onset of Obesity in Sleeve Gastrectomy Candidates
Authors: Robinson Katie , Rowitz Blair , Donovan Sharon M. , Teran‐Garcia Margarita 
Published: June 2021

Sleeve gastrectomy (SG) procedures are becoming increasingly common in response to the rising prevalence of morbid obesity. Post‐operatively, SG patients experience positive outcomes beyond weight loss, including metabolic disease resolution and improved quality of life, which may be impacted by previous life experiences, such as onset and duration of obesity. The degree of influence of these factors is unknown. Therefore, the aim of this research is to describe the characteristics of a cohort of SG candidates and to highlight differences between individuals who reported earlier onset of weight gain and obesity with those who experienced weight gain later in life. Candidates for SG (n=28, 29% male, 46.8 ± 9.0 yrs) completed surveys in which they identified their perceived age of obesity onset and their child body size using a set of child images that represented varying body mass index (BMI) percentiles. Three‐day food logs, physical activity records and body composition via bioelectrical impedance were collected prior to the preoperative diet, on the morning of surgery and one month post‐operatively.At baseline, mean BMI was 48.4 ± 7.4 kg/m2 and body fat percentage was 50.9 ± 5.0%. The most common reported cause of weight gain was poor eating habits (82%), followed by physical inactivity (75%), genetics (46%), stress (36%) and depression (29%). The most common comorbidities were hypertension (54%), hypercholesterolemia (43%), anxiety (36%), type 2 diabetes (32%) and depression (32%). Nine women and two men identified with the child's image that was greater than the 90th percentile (OW). These participants were compared to those who perceived themselves to be normal weight children. Women who reported that they were OW as children had higher BMIs at initial visit (p=0.002). Participants who reported OW as children reported less minutes walking (p=0.05) and no differences in diagnosed comorbidities.Men lost 5.8 ± 1.7 kg and 1.9 ± 0.6 BMI units and women lost 5.5 ± 1.8 kg and 2.0 ± 0.8 BMI units following the two week pre‐operative diet. Men lost significantly more body fat (1.75 ± 2.0 %) than women (0.05 ± 1.3%). Weight loss during pre‐operative diet was not significantly different based on perceived onset of obesity or child body size perception. Although data collection in this cohort is ongoing, our findings suggest that perceived child size and onset of weight gain is associated with adult BMI, however, it does not predict the ability of an individual to lose weight pre‐operatively.Support or Funding InformationThis research was supported by the Agriculture and Food Research Initiative of the USDA National Institute of Food and Agriculture as part of the AFRI Childhood Obesity Prevention Challenge (2011‐67001‐30101) to the Division of Nutritional Sciences at the University of Illinois.
The Influence of Commensal Microflora on Small Intestinal Inflammation and Wnt Pathway in High Fat‐ and Genetically‐induced Obese Apc1638N Mice
Authors: Liu Wei , Crott Jimmy W. , Li Jinchao , Pfalzer Anna C. , Mason Joel B. , Liu Zhenhua 
Published: June 2021

Obesity, which has reached epidemic proportions worldwide, is an established risk factor for some cancers, but the mechanisms responsible for this relationship remain elusive. Our previous study demonstrated that obesity‐induced inflammation elevates Wnt‐signaling, a pro‐tumorigenic pathway that is altered in ~90% of colon cancers. The objective of this study was to examine the role of the intestinal microflora in the increases in inflammation and Wnt‐signaling observed in both high fat‐ and genetically‐induced obese state, and thereby the intestinal tumorigenesis in the Apc1638N mouse model. Three experimental groups with 2 different genotypes and 2 dietary treatments were used: Apc+/1638NLepr+/+ fed a low fat diet (10% fat, Apc‐LFD), Apc+/1638NLepr+/+ fed a high fat diet (60% fat, Apc‐HFD), and Apc+/1638N Leprdb/db fed a low fat diet (Apc‐DbDb). All animals (9~12 mice/group) received experimental diets for 16 weeks from 8 to 24 weeks of age. Because tumors occur exclusively in the small intestine in the Apc1638N model, we examined the microbial composition, inflammatory status and the expressions of Wnt pathway‐specific genes in this organ, as opposed to the large intestine. The relative abundances of 19 bowel cancer associated bacterial taxa were examined by quantitative real‐time PCR. The prevalence of Turicibacter and Desulfovibrio decreased, but Faecalibacterium prausnitizii (F.prausnitizii) increased significantly in high fat diet‐induced obese mice (p < 0.05); whereas in genetically‐induced obese mice, the prevalence of Bifidobacterium, Akkermansia muciniphila (A.muciniphila) and Eubacterium rectale (E.rectale) decreased, but Peptostrptococcus, and Escherichia coli(E.coli) increased (p < 0.05). Both high fat‐ and genetically‐induced obesity increased small intestinal inflammation as indicated by elevated protein levels of IFNγ, IL‐6 or TNF‐α. The changes of inflammatory cytokine transcripts, including TNF‐α, were associated with abundance of Turicibacter, Bifidobacterium, A.muciniphila, E.rectale, and E.coli, as well as the expression of several genes (Wnt5a, Akt, β‐catenin, JNK1, etc.) within the Wnt pathway cascade (p < 0.05). These findings demonstrate that the composition of the small intestinal microflora is affected differentially in high fat diet‐ and genetically‐induced obesity, but both are associated with an elevation of intestinal inflammation and alterations in the expression of Wnt‐signaling related genes.Support or Funding InformationThis project is supported by USDA grants (2014‐67017‐21762, MAS00454), SPHHS Deans Incentive Grant; and the Agricultural Research Service of the United States Department of Agriculture, Cooperative Agreements #58‐1950‐0‐014 and #58‐1950‐4‐003