bluuebunny/crossref_metadata_2025_split huggingface dataset:
Blueberry Diet Derived 3‐(3‐hydroxyphenyl) Propionic Acid (PPA) Suppresses Osteoblastic Cell Senescence to Promote Bone Accretion in Mice
Authors: Chen Jin‐Ran , Lazarenko Oxana P. , Blackburn Michael L. 
Published: June 2021

A blueberry (BB) supplemented diet has been previously shown to significantly stimulate bone formation in rapidly growing male and female rodents. Phenolic acids (PAs) are metabolites derived from polyphenols found in fruits and vegetables as a result of the actions of gut bacteria, and the levels of seven individual PAs were found to be at least 10 times higher in the serum of rats fed a BB‐containing diet compared to those fed a control diet. We have characterized the effects of one such BB‐associated serum PA, 3‐(3‐hydroxyphenyl) propionic acid (PPA), on stimulating osteoblastogenesis. To better understand the mechanistic actions of PPA on bone formation, we administered four doses of PPA (0.1, 0.5, 1 and 5 mg/kg/d) to one‐month‐old female C57BL6/J mice for thirty days. We found that stimulation of osteoblast differentiation and proliferation occurred in PPA treated osteoblastic cells. Using peripheral quantitative CT scan (pQCT) and micro‐CT analyses, we demonstrated differences in bone phenotypes between control and PPA‐treated animals, i.e., bone mass was significantly higher in 1 to 5 mg/kg/d PPA‐treated animals compared to controls (p<0.05). Micro‐CT showed bone volume and trabecular number in 1 mg/kg/d treated animals were 0.14±0.007 and 4.76±0.095 versus 0.12±0.02 and 4.33±0.33 in control animals (p<0.05). The bone formation marker alkaline phosphatase in both serum and bone marrow plasma was dose‐dependently increased in PPA‐treated animals. After aspiration of bone marrow, femur bone protein and RNA were isolated for Western blot and real‐time PCR analysis of signaling transduction. We found that PPA was able to suppress bone senescence signaling as evaluated by measuring senescence‐associated beta‐galactosidase activity, PPARγ, p53 and p21 expression (p<0.05 vs control). In conclusion, blueberry‐derived PPA is capable of altering the mesenchymal stem cell differentiation program and bone cell senescence. Thus, PPA or blueberry‐rich diets merit investigation as potential dietary alternatives to drugs for degenerative bone disorders or to promote bone growth during development. Supported in part by USDA‐ARS Project 6026‐51000‐010‐05S.Support or Funding InformationSupported in part by USDA‐ARS Project 6026‐51000‐010‐05S
Characterization of Anthocyanins in Sweet Potato Shoots
Authors: Su Xiaoyu , Griffin Jason , Katz Benjamin , Tomich John , Wang Weiqun 
Published: June 2021

We previously selected an anthocyanin‐enriched purple‐fleshed sweet potato P40 that prevented colorectal cancer in a murine model. The objectives of this study was to characterize anthocyanins in P40 shoots used as common edible greens. Eight anthocyanins have been identified and quantified in leaves of P40 by HPLC‐MS/MS, whereas fourteen anthocyanins have been found in the control white‐fleshed Bonita and orange‐fleshed Beauregard. The three most abundant anthocyanins were cyanidin 3‐caffeoyl‐p‐hydroxybenzoyl sophoroside‐5‐glucoside, peonidin 3‐caffeoyl sophoroside‐5‐glucoside, and cyanidin 3‐(6″‐caffeoyl‐6″‐feruloylsophoroside)‐5‐glucoside, which comprised up to 58% of total anthocyanins in leaves of P40. The content of anthocyanins in leaves of P40 (17 mg/kg dry matter) was much lower than that in the flesh (13,000 mg/kg dry matter). No anthocyanin was detectable in the stem. The content of anthocyanins in P40 greens was also much lower than that in the Bonita (563 mg/kg dry matter) and Beauregard (334 mg/kg dry matter). The total phenolic content in leaves of P40, as measured by Folin‐Ciocalteu, was 36.8±4.8 mg GAE/g dry weight, compared with Bonita at 46.7±2.1mg GAE/g dry weight and Beauregard at 41.2±5.0 mg GAE/g dry weightSupport or Funding Informationsupported by USDA Cooperative KS511‐1001903
Food‐grade antimicrobial ɛ‐polylysine transiently perturbs the structure of the murine gut microbiome
Authors: You Xiaomeng , Einson Jonah , Lopez‐Peña Cynthia , Song Mingyue , Xiao Hang , McClements D. Julian , Sela David A. 
Published: June 2021

A scientific consensus has emerged that dietary components could guide the establishment and maintenance of microbial communities that colonize the distal gastrointestinal tract. Although ubiquitous in various food systems, it is unknown to what extent that antimicrobial compounds interact with endogenous gut microbiota once consumed. In this study, we investigated the influence of food‐grade biopolymers (i.e. ɛ‐polylysine, pectin, and ɛ‐polylysine‐pectin complexes) on the murine microbiota. Accordingly, CD‐1 mice were fed biopolymers incorporated into a high‐fat diet over a 9‐week time course. Microbial phylogenetic diversity present in fecal extracts were ascertained by high‐throughput amplicon sequencing of the 16s rRNA gene. Moreover, community function was predicted from relative abundances of taxa using PICRUSt. Interestingly, dietary ɛ‐polylysine transiently altered the gut microbiome prior to restoration of the initial microbial population structure at the end of the feeding trial. This is indicative of a community adaptive response to ɛ‐polylysine that is now known to be active against microbes in vivo. Moreover when the cationic ɛ‐polylysine is associated with the anionic polymer pectin, this micro‐bioactive perturbation is mitigated. Finally, the absolute population structure of these murine gut microbial communities was observed to be sex‐dependent, but did not alter the response trajectory to exogenous biopolymers.Support or Funding InformationUSDA
Associations Between Plasma Fatty Acid Composition, Estimated Desaturase Indices, and Measures of Insulin Resistance in 4th‐ and 5th‐grade Children
Authors: Nguyen Lori M , Newman John W , Borkowska Alison G , Zidenberg‐Cherr Sheri , Dharmar Madan , Gray Ira J , Linnell Jessica D , Young Heather M , Scherr Rachel E 
Published: June 2021

Lipidomics is often used to characterize the role of lipids in disease; however, few reports document lipid profiles of children. The aim of this study was to evaluate plasma fatty acid profiles in children and identify potential correlates with markers of metabolic health. The fatty acid profiles of children (n=85, ages 9–12) were determined. Anthropometric data, dietary intake data, and plasma were collected. Fasting plasma was analyzed for total fatty acids, glucose, insulin, and full lipid panels for this cross‐sectional study. Insulin resistance was estimated using calculated HOMA‐IR values. Pearson's correlation was used to exam the relationship between HOMA‐IR and blood biomarkers, analysis of covariance was used to determine differences between groups, and multivariate regression was used to determine the predictors of HOMA‐IR. An increased proportion of palmitic acid (16:0) often characterizes insulin resistant states. A negative correlation between HOMA‐IR values and 18:0/16:0 ratio (r=−0.37, P=0.005) was observed. When analyzed by HOMA‐IR quartiles, the highest quartile had a mean difference of −0.05 in the 18:0/16:0 ratio compared to the lowest quartile (P=0.030). A high stearoyl‐CoA‐desaturase (SCD) index, estimated by taking the ratio of 16:1n7/16:0, has been linked to obesity and metabolic syndrome and in this study HOMA‐IR correlated with SCD‐16 (r=0.35, P=0.001). Mean SCD index was greater among the third and fourth quartiles of HOMA‐IR compared to the first (P=0.020). A low Delta‐5‐Desaturase (Δ5D) is often associated with lower risk for metabolic diseases and mean Δ5D was highest in the fourth quartile of HOMA‐IR (P=0.02). In the linear regression model, BMI, 16:0 ratio, and Δ5D were the strongest predictors of HOMA‐IR (r2=0.38, P=0.01). Consistent with adult studies, insulin resistance in children appears to be associated with increased 16:0 proportions and SCD activity indices in plasma. Though these children appear healthy, the type and level of lipidomic markers may be indicative of the degree of insulin resistance. Evaluating plasma fatty acid profiles of children may help establish early disease risk.Support or Funding InformationFunding provided by University of California Agriculture and Natural Resources Competitive Grant #11‐1018, USDA Training Grant 2011‐38420‐20082, USDA NIFA, Hatch project 221082, USDA Intramural Project 2032‐52530‐022‐00D, and NIH 1U24DK097154‐01. The USDA is an equal opportunity provider and employer.
Identification and Quantification of Carotenoids in Various Phenotypic Sorghum Accessions
Authors: Shen Yanting , Su Xiaoyu , Chen Xi , Rhodes Davina , Katz Benjamin , Tomich John , Wang Weiqun 
Published: June 2021

Sorghum is a staple crop consumed in some regions of Africa and Asia, where vitamin A deficiency is prevalent. The objective of this study was to identify and quantify the carotenoids by LC‐MS in the diversity of phenotypic sorghum accessions. Totally, five carotenoids (α‐carotene, β‐carotene, lutein, zeaxanthin, and β‐crypoxanthin) were identified. Top three carotenoids (lutein, zeaxanthin and β‐carotene) accounted for more than half of the total carotenoids were quantitated. A high content of total carotenoids was detected in the sorghum accessions with yellow endosperm (PI533943, PI655996, PI656096, PI656010), but high β‐carotene content was found in the accessions with brown or yellow pericarp (PI656010, PI655996, PI656096). PI656010 possessed the highest content of both carotenoids and β‐carotene up to 43.7 and 18.6 ng/g dry weight, respectively. No carotenoids were detectable in the accessions with white/red pericarp or white endosperm (PI656112, PI297104). Developing an association of phenotypic diversity of sorghum varieties with carotenoid profile may contribute to a healthy benefit in vitamin A deficiency prevention.Support or Funding InformationSupported by USDA Cooperative KS511‐1001903
Sex‐dependent expression of cytoprotective elements in the kidney in response to a hepatotoxic dose of acetaminophen
Authors: Cooper Daniel , Flores Katiria M , Renfro J Larry , Manautou Jose 
Published: June 2021

Acetaminophen (APAP) overdose can cause liver and kidney necrosis. Male mice are known to be more susceptible to APAP hepatotoxicity than females, suggesting that gender‐related genotypic traits play a role in APAP toxicity. Gender‐related differences in susceptibility to APAP nephrotoxicity in mice are also well documented and attributed to differential renal expression of CYP2E1. Recent studies in our laboratory investigated the role of the transcription factor nuclear factor erythroid 2‐related factor 2 (Nrf2), the efflux transporter multidrug resistance‐associated protein 4 (Mrp4), and the drug metabolizing enzyme flavin‐containing mono‐oxygenase 3 (Fmo3) in sex‐dependent susceptibility to APAP‐induced hepatotoxicity in mice. However, the expression and potential role of Nrf2, Fmo3, and Mrp4 in the responsiveness of male and female mouse kidneys to toxic doses of APAP has not been investigated. In the current study, we examined gene expression and protein levels of Nrf2, Mrp4, and Fmo3 in the kidneys of APAP (400 mg/kg) and vehicle treated male and female mice. Plasma alanine aminotransferase (ALT) and blood urea nitrogen (BUN) levels were measured to assess liver and renal damage. Confirmation of tissue damage was made by histopathological analysis. Immunoblots and qRT‐PCR analysis revealed that basal renal Fmo3 gene and protein expression is significantly greater in male than female mice, while Mrp4 expression is higher in females. No differences in basal Nrf2 gene and protein expression were observed among genders. High plasma ALT values and histopathological analysis indicated the presence of hepatotoxicity in both males and females receiving APAP. Renal damage was not evident, as BUN values were within the normal range for both genders and the kidney histology was normal. In response to APAP treatment, no changes in renal expression of Mrp4 or Fmo3 were observed in either gender, consistent with the absence of renal pathology. However, renal Nrf2 gene and protein expression decreased significantly in female mice after APAP dosing. In conclusion, this study documents for the first time the difference in basal renal expression levels of Nrf2 and Fmo3 in male and female mice. Further studies will investigate the expression and regulation of these genes at doses of APAP resulting in renal oxidative stress and toxicity and their potential involvement in the sex‐dependent responses to APAP induced renal damage.Support or Funding InformationSupported by NSF and NIH
Effects of NOX‐1 on Real‐Time Blood Nitric Oxide and Hydrogen Peroxide in Acute Hyperglycemia
Authors: Mawhinney Ashley Elizabeth , Bertolet Michael , Minni Michael , Barsotti Robert , Young Lindon , Chen Qian 
Published: June 2021

Hyperglycemia has been associated with vascular endothelial dysfunction in part by a reduction in nitric oxide (NO) production and increased oxidative stress (e.g., increased superoxide (SO) and hydrogen peroxide (H2O2). Endothelial‐derived NO can be significantly reduced by increased SO/H2O2 in part by the activation of NADPH oxidase during hyperglycemia. Of the 7 NADPH oxidase isoforms, NOX1 is mainly expressed in the vasculature and may play a major role in hyperglycemia induced oxidative stress and vascular endothelial dysfunction. To test this hypothesis, we measured blood NO and H2O2 levels in real time via NO and H2O2 microsensors inserted into femoral veins of rats. Hyperglycemia (e.g., 200 mg/dl) was maintained by infusion i.v. of 30% glucose solution for 3 hours with or without a selective NOX1 inhibitor, ML171. We found that hyperglycemia for 3 hours significantly increased blood H2O2 levels by 1.88±0.4 μM (n=6) compared to the saline infused control (P<0.05, n=2). By contrast, ML171 (1 and 5 μM) reduced hyperglycemia‐induced H2O2 levels by 1.36±0.61 μM (n=8) and 4.35±1.02 μM (n=5), respectively, at the end of experiment. Meanwhile, hyperglycemia significantly reduced blood NO levels by 82.48±38.12 nM (n=3) compared to the saline control (P<0.05, n=2). By contrast, ML171 (1 μM) attenuated the hyperglycemia induced decrease in blood NO levels and increased blood NO levels by 71.15±24.00 nM (n=5) at the end of experiment. Our preliminary results indicate that NOX1 activation may contribute to hyperglycemia‐induced oxidative stress and NO reduction. Furthermore, inhibition of NOX1 may mitigate the deleterious effects of hyperglycemia.Support or Funding InformationThis study was supported by Division of Research, the Center for Chronic Disorders of Aging, and Department of Bio‐Medical Sciences at Philadelphia College of Osteopathic Medicine.
A Sex‐Specific Role for Egr1 in Mediating Norepinephrine‐Induced Contraction in Mesenteric Arteries
Authors: Thakali Keshari M , Wahl Elizabeth C , Kang Ping , Shankar Kartik 
Published: June 2021

The immediate early transcription factor early growth response 1 (Egr1) is upregulated in smooth muscle cells of atherosclerotic plaques and in adipose tissue of obese animals and humans. In a heterologous endothelial cell expression system, Egr1 overexpression upregulated the α2δ accessory subunit of the L‐type voltage gated Ca2+ channel (Fu et al, Gene; 2003). Moreover, Ca2+ influx via voltage‐gated Ca2+ channels increased Egr1 expression, suggesting that Egr1 regulates ion channel flux and vascular tone. In light of these findings, we tested the hypothesis that mesenteric arteries (MA; 2nd order) from Egr1 knockout (KO) mice would exhibit reduced contractility compared to MA from their wild‐type (WT) litter mates. Isolated vessel contractility was assessed using wire myography (DMT 620M). A small rightward shift in cumulative concentration response curves to norepinephrine (NE, 10−9 – 10−5M) in MA from male KO mice compared to WT littermates (−log EC50 = 5.96±0.11 vs −5.68±0.03 M NE, respectively; p=0.08) was observed, while there was no difference in NE‐induced contraction in female KO and WT mice. Neither maximal NE‐induced constriction nor maximal acetylcholine‐induced relaxation in NE‐preconstricted MA was different between male KO and WT mice or female KO and WT mice. Current studies are leveraging RNAseq to assess global transcriptome profiles of aorta from male and female WT and KO mice, to determine which pathways are impacted by Egr1. Our preliminary arterial contractility data suggest that smooth muscle Egr1 plays a role in modulating vascular tone.Support or Funding InformationThis work was funded by USDA‐ARS Project 6026‐51000‐010‐05S.
Base Excision Repair of Bulky DNA Adducts Generated by the Antitumor Drug Yatakemycin
Authors: Mullins Elwood A. , Shi Rongxin , Igarashi Yasuhiro , Eichman Brandt F. 
Published: June 2021

Threats to genomic integrity are mitigated by DNA glycosylases, which initiate the base excision repair pathway by locating and excising aberrant nucleobases. A hallmark of these and other DNA repair enzymes is their use of base flipping to sequester modified nucleotides from the DNA helix and into an active site pocket. Consequently, base flipping is generally regarded as an essential aspect of lesion recognition and a necessary precursor to base excision. We recently described the first DNA glycosylase mechanism that does not require base flipping for either binding or catalysis1. The DNA glycosylase AlkD recognizes aberrant base pairs through contacts with the phosphoribose backbone, while the damaged nucleobase remains stacked in the DNA duplex, and and uses catalytic CH–π and charge–dipole interactions to preferentially stabilize the transition state. We now show through a combination of crystallographic, biochemical, biophysical, and cellular techniques how this unique mechanism enables AlkD to repair large adducts formed by yatakemycin (Fig. 1), a member of the duocarmycin and CC‐1065 family of antimicrobial and antitumor natural products. Bulky adducts of this, or any type, are not excised by DNA glycosylases that use a traditional base‐flipping mechanism. Hence, these findings represent a new paradigm for DNA repair and provide insights into damage recognition and base excision.Support or Funding InformationThis work was funded by the National Science Foundation (MCB‐1122098 and MCB‐1517695) and the National Institutes of Health (R01ES019625).
Transcriptional and Post‐transcriptional Regulatory Mechanisms Lead to Metabolic Adjustment in Response to Glucose Availability
Authors: Munshi Azad Hossain , Claggett Julia M , Johnson Tracy L. 
Published: June 2021

The transcription factor Gcr1 controls expression of over 75% of the genes in actively growing yeast. Yet, despite its widespread effects, regulation of Gcr1 itself remains poorly understood. Here we show that posttranscriptional Gcr1 regulation is nutrient‐dependent. Moreover, GCR1 RNA contains a highly conserved intron, which allows the cell to generate multiple RNA and protein isoforms whose levels change upon glucose depletion. Intriguingly, an isoform generated by intron retention is exported from the nucleus, and its translation is initiated from a conserved, intronic translation start site. Expression of gene products from both the spliced and unspliced RNAs is essential, as cells expressing only one isoform cannot adjust their metabolic program in response to environmental changes. Finally, we show that the Gcr1 proteins form dimers, providing an elegant mechanism by which this one gene, through its regulation, can perform the repertoire of transcriptional activities necessary for fine‐tuned environmental response. So, this study provides a striking example of how alternative splicing in the form of intron‐retention can present the cell with novel mechanisms for regulating gene expression.Support or Funding InformationThis work was supported by the National Science Foundation (MCB‐1051921 to T.L.J.) and the National Institute of General Medical Sciences ( GM‐085474 to T.L.J.)
Vacuolar Turnover of rRNA Mediated by An Autophagy‐Dependent Mechanism Is Necessary To Maintain Cellular Homeostasis
Authors: MacIntosh Gustavo C , Floyd Brice , Morriss Stephanie , Bassham Diane 
Published: June 2021

Ribosomes are major components of all cells. Much is understood about their biosynthesis and several pathways have been characterized for the elimination of misassembled ribosomal subunits. However, little is known about the decay of stable ribosomes and recycling of rRNA. RNase T2 ribonucleases represent one of the most widely distributed RNase families, and they are conserved in all eukaryotes, many prokaryotes, and some viruses. Our work showed that RNS2, a class II RNase T2 in Arabidopsis thaliana, is essential for normal ribosomal RNA decay. Null rns2‐2 plants show constitutive autophagy, RNA accumulation in the vacuole, and longer rRNA half‐life. Autophagosomes and autophagic bodies in rns2‐2 mutants contain RNA and ribosomes, suggesting that autophagy is activated as an attempt to compensate for loss of rRNA degradation. Total RNA levels increase in the rns2‐2, in the autophagy mutants atg9‐4 and atg5‐1 which are deficient in core autophagy machinery, and in rns2‐2atg9‐4 and rns2‐2atg5‐1 double mutants. Interestingly, rns2‐2atg5‐1 had significantly higher total RNA than the single mutants and the rns2‐2atg9‐4 double mutant. These results suggest that, together with RNS2, autophagy has a role in RNA turnover. In addition, rRNA accumulates in the vacuole in rns2‐2 mutants. Vacuolar accumulation of rRNA was blocked by disrupting autophagy via an rns2‐2atg5‐1 double mutant but not by an rns2‐2atg9‐4 double mutant. Furthermore, atg5‐1 vacuoles had significantly lower amounts of rRNA than WT or atg9‐4. Thus, it is clear that ATG5 and ATG9 have different roles in the turnover of rRNA. Our results suggest that an ATG5‐dependet selective autophagic process transport rRNA to the vacuole for degradation by RNS2. Disruption of this process in the rns2‐2 mutant triggers general macroautophagy which depends on ATG9 and ATG5, as a compensatory mechanism to continue the regular turnover of ribosomes and to re‐establish cellular homeostasis.Support or Funding InformationThis work was supported by grant no. MCB‐1051818 from the United States National Science Foundation to G.C.M. and D.C.B.