bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cell morphology changes in 3T3‐L1 cells grown in monolayer and treated with cinnamon extract
Authors: Baxendell Kayla , Aulthouse Amy , Stockert Amy 
Published: April 2020

Cinnamon has long been used medicinally and has gained attention more recently due to its ability to decrease blood glucose values in type 2 diabetes. Research has suggested potential targets in the cell responsible for the metabolic effects observed with cinnamon, however, a collective mechanism of action is still unknown. This study demonstrates morphological changes observed in 3T3‐L1 cells grown in monolayer and treated with cinnamon extract that mimic cell differentiation observed in some three‐dimensional models. Data is presented that demonstrates changes in cell shape with increasing concentrations of cinnamon extract. Evidence of lipid accumulation is visualized by oil red O staining as early as 0.5 weeks following cinnamon treatment. Monolayer cells exhibit round morphology when treated with cytochalasin D, an actin polymerization inhibitor. Together these data suggest that one potential metabolic target of cinnamon extract may result to alter actin polymerization leading to cell rounding. The morphological change from flat to round cells potentially stimulates changes in gene expression that lead to differentiation. Future work will determine the extent of differentiation observed in the cinnamon treated cells compared to those grown with cytochalasin D and those grown in three‐dimensional culture. This study outlines possible unique therapeutic approachs for the alteration of cell differentiation that could result in desired metabolic changes.Support or Funding InformationOhio Northern University Raabe College of Pharmacy and the Department of Biological and Allied HealthThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gender specific effects of Nr4a1 on functional β‐cell mass
Authors: Herring Jacob A. , Smith Courtney J. , Kener Kyle B. , Wynn Adam G. , Austin Matt C. , Tessem Jeffery Sivert 
Published: April 2020

Nuclear hormone receptor4a1 (Nr4a1) plays a critical role in β‐cell replication, function, and survival. β‐cell replication is tightly regulated, most β‐cells in the body are formed during development and shortly after birth. Nr4a1 full body knock‐out mice have a 50% reduction in total β‐cell area. Nr4a1 knock down INS‐1 832/13 β‐cells have decreased mitochondrial respiration, glucose stimulated insulin secretion and cellular proliferation. Fatty acids regulate Nr4a1 activity and expression. Here we present the effects of β‐cell specific Nr4a1 deletion on mice exposed to a high fat or normal chow diet beginning at 3‐months of age. Our data demonstrate that loss of β‐cell Nr4a1 results in improved non‐fasting blood glucose and glucose tolerance in male mice fed a high fat diet for 4 months. Interestingly, female mice deficient for Nr4a1 have impaired glucose tolerance as early as 1 months of high fat feeding. Furthermore, we show data regarding β‐cell mass and glucose stimulated insulin secretion. These data demonstrate a clear gender bias in Nr4a1 function in the β‐cell.Support or Funding InformationADA 1‐17‐IBS‐101 to JSTThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Potential Role of Exosomes in Countering Vedolizumab‐Based Therapy in IBD Patients
Authors: Domenis Rossana , Cifù Adriana , Fabris Martina , Scardino Giulia , Zilli Maurizio , Marino Marco , Curcio Francesco 
Published: April 2020

IntroductionInflammatory bowel diseases (IBD), including Crohn's disease and ulcerative colitis, are chronic and debilitating inflammatory disorders of the gastrointestinal tract. In the development of new treatment options, it was realized that the attenuation of lymphocyte translocation into the inflamed gut tissue results in a reduction in local inflammation and thus decrease IBD severity. Vedolizumab (VDZ) is a monoclonal antibody targeting α4β7 integrin, which is expressed specifically by a subset of gastrointestinal‐homing T‐lymphocytes. Although VDZ showed promising results in various clinical studies, in common with all existing IBD therapies, a significant number of patients do not respond and validated markers and mechanistic insights to predict the populations that will derive sustained benefit are currently lacking. We hypothesized that circulating exosomes express on their surface high levels of α4β7 integrin, which could bind VDZ and interfere with its activity and therapeutic efficacy.MethodsExosomes were isolated from serum of blood donors and VDZ‐treated patients by polymer‐based precipitation (Exoquick), analysed for concentration (Exocet) and validated for exosomal markers expression. The surface expression of α4β7 integrin was evaluated by flow cytometry on exosomes‐bound beads. The levels of exosome‐bound VDZ were investigated by Promonitor‐VDZ ELISA kit and western blot analysis. Finally, exosomes isolated from blood donor's serum were incubated with increasing concentration of VDZ and then exosomal‐bound VDZ levels were analysed.ResultsFlow cytometry analysis revealed that serum exosomes, either from the patients and from the donors, express high levels of the VDZ target, α4β7 integrin. A significant VDZ levels were measured in exosomes purified from VDZ‐treated patient's serum exosomes. Of note, we found that exosomes purified from blood donor's serum were able to bind VDZ in dose‐dependent manner.ConclusionsOur data suggest that circulating serum exosomes bind VDZ, as they express on the surface the target integrin. Accordingly, exosomes might contribute to drug sequestration, possibly affecting the therapeutic efficacy of VDZ in IBD patients. Further studies are needed to define the possible correlation between VDZ exosomal sequestration and patient's response.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sex differences in anxiety‐like behaviors following chronic intermittent ethanol and/or repeated stress exposures using an adult zebrafish (Danio rerio) model
Authors: Devaud Leslie , Johnson Heather , Lally Jessica , Perez Erica 
Published: April 2020

Studies on ethanol use have found that stress often impacts responses, such that it may promote increased ethanol intake and, therefore, risk for harm. Additional evidence suggests that stress, especially severe stress, may have a greater negative effect from ethanol in females than males. To improve our understanding of these potential interactions, we used an adult zebrafish model to assess behavioral activities in 6 mth old male and female zebrafish following treatment with chronic intermittent ethanol exposure and/or repeated stress exposure. The 4 treatment conditions were: control (tanks moved to the testing bench but no other manipulations), twice‐daily 60 min 0.5% ethanol exposure and/or 3 times daily 40 sec stress exposures for a total of five days. We used a custom light/dark tank to assess behaviors at approximately 16 hr after the final treatments. Zebrafish were tested individually with data collected for a total of 8 min every 10 seconds: (1) light vs. dark preference, (2) crossings between light and dark, (3) preference for tank top vs. bottom and (4) general activity levels (normal, hyperactive or freezing). Time spent in each activity was summated across the testing period for each animal and averaged by sex within treatment condition. Overall, control females spent more time in the dark (20%) compared to males, suggesting a greater sensitivity to the novelty of the testing tank for female than male fish. We observed a large increase in time spent in the dark for both males (34%) and females (59%) compared to control values for the EtOH‐only treatment whereas the stress + EtOH condition values were similar to controls. There was a prominent sex difference in hyperactivity with the stress only males showing a 26% reduction compared to control levels and females having a 38% increase compared to males. Interestingly, we also saw a reduction (17%) in hyperactivity for EtOH only males compared to control values but a large increase (40%) for the ethanol only females. Again, the stress + EtOH condition values were similar to controls for both sexes, suggestive of some resiliency when exposed to both the intermittent EtOH as well as the repeated stress procedure. We also noted less freezing (believed to reflect high levels of fear/anxiety in the fish) in males and females that were treated with the repeated stress, suggesting that this helped the fish adapt to the novelty of the testing tank more than for the other treatment conditions. These data support the utility of zebrafish as an animal model to study the impact of stress on ethanol responses and showed that behavioral responses support an interaction of stress with ethanol by sex.Support or Funding InformationSupported by Pacific University School of Pharmacy RIG (LLD)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Determining the Molecular Characteristics of How Ligands Interact with an ABC Transporter
Authors: Gose Tomoka , Fukuda Yu , Das Sourav , Wang Yao , Lynch John , Shelat Anang , Schuetz John D. 
Published: April 2020

ABC transporters have key roles in cellular response to chemotherapy and drug bioavailability. The engagement of drugs and other ligands with ABC transporters is the crucial first step in the transport cycle. For mammalian ABC transporters, conventional knowledge regards the high affinity ligand‐binding state as when ATP is not bound to the ABC transporter; however, in vivo, this circumstance would be rare as the ATP concentration in most cells is approximately 5 mM. Using ABCG2 and the important class of drugs (tyrosine kinase inhibitors) we performed a thermal shift assay using two models (intact cells and isolated membranes) to investigate how these ligands engage ABCG2. We discovered, unexpectedly, that drug binding to ABCG2 can be either ATP‐independent, or ATP‐dependent. These findings run counter to the prevailing concept that ligands engage ABC transporters with high affinity when ATP concentrations are low and are reduced when ATP concentrations are high. Further, for those ligands requiring ATP for binding, ATP‐binding (not hydrolysis) appears to be the sole determinant because an ABCG2 mutant incapable of hydrolyzing ATP appeared to have little effect on ligand binding. For the ATP‐independent ligand, the EC50 of binding was minimally affected by ATP, while the affinity and maximum binding of the ATP‐dependent ligand was markedly enhanced by the presence of ATP. Using a non‐hydrolyzable ATP‐coupled bead as a probe for ATP binding, further studies showed that the binding to ABCG2 of this ATP mimetic was strongly influenced by the ligand, but only when the ligand's binding was impacted by ATP, thus suggesting communication occurs between the nucleotide binding domain and the ligand binding site. Previous studies have suggested ATP lowers the binding affinity for ligands. This contrasts with the current findings showing some ligands require ATP for ligand binding, while others do not. This has important mechanistic implications for interpreting not just ABC transporter function, studied in dynamic systems, but also for the development of ABCG2 inhibitors and understanding substrate interactions.Support or Funding InformationThis work was supported by NIH and by ALSAC.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Absence of Angiotensin ll type 2 (AT2) Receptors Increases Airway Inflammation and Responsiveness in an Allergic Mouse Model of Asthma
Authors: Bhavsar Riyakumar , Rajalingam Sahith , Patel Mehaben , Prasad Praveena , Ponnoth Dovenia S 
Published: April 2020

Angiotensin ll (Ang ll) is produced locally in the lungs and may potentiate airway responsiveness in asthmatic patients. Ang ll induces its effects by activating two GPCR subtypes: Ang II receptor type‐1 and type‐2 (AT1 and AT2). This lab has previously found that AT2 receptor agonist plays a significant role in reducing airway inflammation and reactivity (FASEB J April 2018 32: 829.6). For the current study, we characterized the role of AT2 receptors in airway hyperreactivity and inflammation using AT2 receptor knockout (AT2KO; Hein L, Barsh et. al. Nature. 1995;377:744–747) and corresponding wild‐type (WT) mice that were divided into two experimental groups each: control (CON) and allergen sensitized‐challenged (SEN). Mice were sensitized (i.p.) on days 1, 6 with 0.2μg ovalbumin (OVA) followed by 5% OVA aerosol challenges on days 11–13. Whole body plethysmography (measuring airway responsiveness as enhanced pause, Penh) and bronchoalveolar lavage (BAL) studies were performed. Methacholine (MCh; 48mg/ml) produced significantly higher airway responsiveness in AT2KO SEN mice compared to WT SEN (324.7±66% in AT2KO SEN vs 166±56.3%, p<0.05). Total cell count analysis showed increased number of cells in AT2KO SEN (3.3±0.2 × 106 vs. 1.4±5 × 106 in WT SEN, p<0.001). Differential BAL cell analysis showed increased eosinophils (73±8% in AT2KO SEN vs. 50.11±2.84% in WT SEN, p<0.05). Macrophages were lower in AT2KO SEN than WT SEN (15±4% vs. 40±3%; p<0.001). These data indicate that absence of AT2 receptors increases airway inflammation and responsiveness in our model of asthma.Support or Funding InformationInstitutional Research Grant Long Island UniversityThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cholesterol regulates cardiac KATP subunit expression through an SREBP‐dependent mechanism
Authors: Geiger Robert , Fatima Naheed , Schooley James , Flagg Thomas 
Published: April 2020

We recently discovered that the histone deacetylase inhibitor, trichostatin A (TSA), increases the expression of the sulfonylurea receptor 2 (SUR2; Abcc9) subunit of the ATP‐sensitive K+ (KATP) channel in HL‐1 cardiomyocytes. Importantly, the effects of TSA were abolished if cells were treated with exogenous cholesterol, suggesting that cholesterol may suppress channel expression. In the present study, we tested the hypothesis that TSA increases SUR2 by depleting cholesterol and activating the sterol response element binding protein (SREBP) family of transcription factors. As expected, treatment of HL‐1 cardiomyocytes with TSA (30 ng/mL) caused a time dependent increase in SUR2 mRNA expression that correlates with the time course of cholesterol depletion assessed by filipin staining. This is specific for SUR2 and we observe little or no change in expression of the SUR1 subunit. Consistent with a central role for cholesterol‐dependent activation of SREBP in driving the increase in SUR2 mRNA expression, we observe a marked increase in the cleavage of SREBP1 following TSA, and this is inhibited by addition of cholesterol to the media. Further supporting the role of SREBP following TSA treatment, we observe increased expression of other SREBP‐dependent genes including PCSK9 and inducible degrader of LDLr (IDOL). Lastly, in luciferase promoter assays SREBP‐1a markedly increased reporter gene expression driven by the upstream SUR2, but not SUR1, promoter. Taken together, our data are consistent with the conclusion that cholesterol depletion by TSA leads to activation the SREBP signaling pathway that directly acts on the SUR2 promoter to increase gene expression. This finding suggests a novel means by which hypercholesterolemia may exert negative effects on the cardiovascular system by suppressing expression of the KATP channel.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of chondroitin sulfotransferases (CHST3,7,11,15) and chondroitin sulfatases in normal and malignant prostate stroma and epithelium
Authors: Tobacman Joanne Kramer , Feferman Leo , Bhattacharyya Sumit 
Published: April 2020

Previous work demonstrated marked decline in arylsulfatase B (ARSB; N‐acetylgalactosamine‐4‐sulfatase) in malignant prostate tissues by immunohistochemistry, activity assay, and mRNA expression. In contrast, activity of galactosamine‐N‐acetyl‐6‐sulfatase (GALNS; N‐acetylgalactosamine‐6‐sulfatase) was significantly increased in malignant prostate tissues. These findings were associated with marked increase in chondroitin 4‐sulfate (C4S), decline in chondroitin 6‐sulfate (C6S), and increased C4S:C6S ratio in the malignant tissues. To further evaluate the chondroitin sulfation, mRNA expression of chondroitin sulfotransferases and chondroitin sulfatases was detected in malignant and normal prostate stromal and epithelial tissues obtained by laser capture microdissection (LCMD) of prostate tissues obtained from the Biorepository of the University of Illinois at Chicago. Primers were identified using Primer3, and QPCR was used to determine expression levels. Confirmation of epithelial vs. stromal content was confirmed by comparative expression of vimentin and E‐cadherin. ARSB expression was similar in the control and malignant stromal tissue and more than two‐fold higher than in the normal epithelial tissue. ARSB expression was reduced by over 50% in the malignant epithelium compared to normal epithelium. In contrast, GALNS expression was over 50% less in the stromal tissues than in the epithelium, and was increased almost two‐fold in the malignant vs. normal prostate epithelium. The expression of CHST15, which adds a 6‐sulfate group to C4S, was increased over four‐fold in the malignant prostate epithelium, and significantly reduced in both normal and malignant stromal tissue. Expression of the 6‐O sulfotransferases CHST3 and CHST7 was also determined. Normal and malignant stromal CHST3 was about 50% the level in the normal and malignant epithelium. CHST7 was slightly less in the malignant epithelium, compared to the normal epithelium and the normal and malignant stroma. The 4‐O sulfotransferase CHST11 was reduced about 50% in the malignant epithelium, compared to normal epithelium, and increased about 80% in the normal and malignant stroma. These findings suggest that increased expression of CHST15 and GALNS in the epithelium may lead to enhanced production of C4S. This is perhaps compensatory, due to decline in CHST11. However, in association with decline in ARSB, these effects lead to increased epithelial C4S. Since increased C4S has been associated with reduced activity of SHP2, the tyrosine‐protein phosphatase non‐receptor type 11 (PTPN11) and increased availability of galectin‐3, these changes have important consequences for cell signaling, growth, and metabolism and for stromal‐epithelial interactions.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endothelium‐specific disruption of IGF‐1 signaling impairs blood flow regulation in mice
Authors: Tarantini Stefano , Fulop Gabor A , Yabluchanskiy Andriy , Kiss Tamas , Balasubramanian Priya , Csipo Tamas , Lipecz Agnes , Sonntag William E , Csiszar Anna , Ungvari Zoltan 
Published: April 2020

Circulating levels of IGF‐1 decline during aging in humans, which significantly increases the risk for cardiovascular diseases and promotes cerebromicrovascular impairment. To test the hypothesis that impaired IGF‐1 input to endothelial cells promote aging‐like microvascular dysfunction, we utilized a novel mouse model: Igfr1f/f VE‐Cadherin‐Cre mice. We found that endothelium‐dependent increases in cerebral blood flow in response to contralateral whisker stimulation (laser speckle contrast imaging) were attenuated in mice with endothelium‐specific disruption of IGF‐1 signaling. In control mice functional hyperemia was significantly decreased by pharmacological inhibition of NO synthesis (L‐NAME). In contrast, increases in cerebral blood flow in response to contralateral whisker stimulation were unaffected by L‐NAME in Igfr1f/f VE‐Cadherin‐Cre mice. Collectively, our findings support the concept that IGF‐1 signaling is essential for healthy cerebromicrovascular endothelial function and provide evidence that decreased IGF‐1 input to endothelial cells results in impaired NO mediation of functional hyperemia, which may contribute to cognitive decline associated with age‐related IGF‐1 deficiency.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Age Reduces Autophagy and Mitochondria Related Gene Expression in Human Skeletal Muscle
Authors: Wu Hexirui , Dridi Sami , Baum Jamie , Ferrando Arny , Wolfe Robert 
Published: April 2020

In the US, the population of older adults over 65 reach 98 million by 2060. Aging is associated with increased risk for multiple chronic diseases such as obesity, cardiovascular diseases, and sarcopenia (age‐related muscle loss). Skeletal muscle is a major site of metabolic activity and the most abundant tissue in the human body. Mitochondria play a key role in ensuring adequate levels of ATP needed for skeletal muscle contraction. Moreover, when subcellular organelles are found to be dysfunctional or the cells discover themselves in a critical state, they will start a recycling process called autophagy to ensure survival. Autophagy that recycles damaged or dysfunctional mitochondria is called mitophagy. In skeletal muscle from elderly, there is lower ratio of healthy mitochondria versus damaged mitochondria. This could be caused by reduced autophagy. The objective of this study was to determine if there was an effect of age on the genes related to mitochondrial function, autophagy and mitochondrial biogenesis in skeletal muscle from young and old participants. Muscle biopsies from vastus lateralis were taken from young (n=23, thirteen males and ten females, ages 29.7±1.02) and old (n=20, ten males and ten females, ages 65.6±4.29) adults. Gene expression related to autophagy and metabolism was measured using real‐time PCR. The expression of autophagy‐related genes such as p62, ATG5, Atrogin1, Beclin1, LC3B and MURF1 was lower in old subjects compared with their young (p<0.05). Two genes, ATF4 and GADD54A, involved in cell survival were decreased in old subjects compared to young (p<0.05). Genes relate to mitochondrial functions, UCP2, NRF1, PGC1α, PPARγ, and Tfam, showed no changes with age. Genes related to fatty acid metabolism, ACC, FATP1, FATP4, PPARα, SREBP1, were down‐regulated in old subjects (p<0.05). In conclusion, these data suggest dysfunctional autophagy could be the reason for mitochondrial damages found in older skeletal muscle in other reports, and the compromise in fatty acid metabolism which leads to intramuscular fat accumulation.Support or Funding InformationArkansas Biosciences InstiuteThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.