bluuebunny/crossref_metadata_2025_split huggingface dataset:
The P2Y2 Receptor Mediates Hyperglycemia‐induced Insulin Resistance in Human Skeletal Muscle Cells
Authors: Nasrullah Mohammed Zahid , Peng Qianman , Shen Jianzhong 
Published: June 2021

ObjectiveIt has been known that insulin resistance could lead to hyperglycemia and diabetes; however, the exact cause effect relationship between hyperglycemia and insulin resistance is not fully understood. Since over nutrition often leads to energy excess and ATP is the major cellular energy source, we hypothesized that abnormal high level of extracellular glucose promotes skeletal muscle cells to release ATP, which in turn activates the purinergic P2Y2 receptor, resulting to insulin resistance.Methods and ResultsReal‐time RT‐PCR analysis indicated that the P2Y2 receptor mRNA was significantly up‐regulated during human skeletal muscle cell differentiation, and functional P2Y2 receptor expression was confirmed by measuring intracellular [Ca2+]i mobilization in response to ATP or UTP in mature human skeletal muscle cells (HSKMC). Interestingly, incubation of HSKMC with high glucose (25 mM) triggered significant amount of ATP release as compared with normal glucose (6.1 mM). In addition, both ATP and UTP dramatically suppressed insulin‐induced AKT activation in a dose‐dependent manner in HSKMC, which could be reversed by AR‐C118925, a selective P2Y2 receptor antagonist, suggesting a role of P2Y2 receptor in control of insulin receptor signaling. The P2Y2 receptor suppression of the AKT pathway was selective, since ATP/UTP treatment potentiated insulin‐induced ERK1/2 pathway in HSKMC. Furthermore, we found that activation of the P2Y2 receptor significantly inhibited insulin‐induced glucose uptake in HSKMC.ConclusionsWe concluded that ATP release and P2Y2 receptor activation is a previously unrecognized cellular signaling node linking hyperglycemia and insulin resistance in HSKMC through the PI3K‐AKT pathway, highlighting that P2Y2 receptor may be a new drug target for the treatment of diabetes and relevant metabolic diseases.Support or Funding InformationSupported in part by NIH grant 1R01HL125279‐01A1 (to J.S.)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
USP Potency Equated Enriched Bovine and Porcine Mucosal Heparins Exhibit Comparable Biochemical and Pharmacological Effects
Authors: Baig Nausheen , Kouta Ahmed , Siddiqui Fakiha , Farooqui Ambar , Hoppensteadt Debra , Jeske Walter , Iqbal Omer , Fareed Jawed 
Published: June 2021

ObjectiveCommercially available native porcine (NP) heparins exhibit a higher potency (200 U/mg) in contrast to their native bovine (NB) counterpart (140 U/mg). Enriched versions of porcine heparins (EP: 240 U/mg) and bovine heparins (EB: 180 U/mg) were developed using improved separation methods. At a gravimetric concentration, porcine heparins (NP and EP) exhibit stronger effects in contrast to bovine heparins. The purpose of this study is to compare the biologic properties of these agents at equated USP potencies.MethodsNP, EP, NB, and EB heparins were obtained from Yino Pharma (China). A commercially available porcine mucosal heparin was obtained from Hospira/Pfizer (USA) (190 U/mg). Working dilutions were prepared at potency equated levels of 10 U/mL. Whole blood aPTT and TT were measured on the ST4 instrument at a concentration range of 0–1 U/mL. Whole blood ACT was measured using a Hemochron instrument at a fixed concentration of 1 U/mL. TT, aPTT, and protamine neutralization profiles were also measured in plasma supplemented with each agent at a concentration range of 0–1 U/mL. The USP potency assay was carried out using chromogenic antiprotease assays (Biophen Kits). The inhibitory concentrations at 50% (IC‐50) were measured using a purified antithrombin biochemical system employing a kinetic method. Thrombin generation inhibition assay was carried out using a CAT method (Diagnositca Stago, Paris, France).ResultsIn the whole blood ACT method, all drugs produced comparable effects with values between 200–250 seconds. In the whole blood aPTT and TT assays, all results were comparable. In the plasma supplementation studies, all drugs were comparable in the aPTT, factor Xa, and factor IIa studies and the protamine neutralization profiles. However, in the TT assay, NB and EP were somewhat weaker. The anti‐Xa and anti‐IIa activities of all drugs were comparable. The IC‐50 values for the anti‐Xa ranged from 0.33–0.40 u/mL and the anti‐IIa values ranged from 0.38–0.42 U/mL. In the thrombin generation assays, the EB heparins showed slightly weaker activities.ConclusionThese results show that at potency equated concentrations, both the native and enriched porcine and bovine heparin preparations exhibit comparable biochemical and anticoagulant responses. Therefore, the hypothesis that potency equated native/enriched bovine and porcine heparins exhibit comparable biochemical and anticoagulant activities is validated.Support or Funding InformationNoneThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Acai fruit extracts and anthocyanins alter osteoblast proliferation and the expression of HDAC1 and osterix/Sp7 in vitro and in vivo
Authors: Mahady Gail B , Nishikant Raut , Lawal Temitope O 
Published: June 2021

Musculoskeletal injuries and disorders are the leading cause of disability in the United States and affect more than one out of every two persons in the United States age 18 and over, and nearly three out of four persons aged 65 and over. Bone health is critical to the overall health and quality of life for Americans, and greater efforts are needed to address the burdens associated with osteoporosis and related bone diseases. Evidence from a variety of studies strongly points to a positive link between fruit and vegetable consumption and indices of bone and joint health, although the mechanisms are not well understood. In this work we investigate the effects of Acai fruit extract (AFE) and anthocyanidins on osteoblastogenesis in vitro and in vivo. hFOB human osteoblasts were obtained from ATCC and cultured in 1:1 mixture of Ham's F12 Medium Dulbecco's Modified Eagle's medium, with 2.5 mM L‐glutamine (without phenol red). An extract of acai fruits (AFE) was prepared in methanol and purified cyanidin‐3‐glucoside were purchased. Double transgenic medaka (dT; osterix/Sp7:mCherry were maintained at UIC under protocol #17‐166. Larvae at 9–12 dpf were staged and then treated for 5 days with the extracts and compounds (1–10 mg/ml). Osteoblast formation and distribution were analyzed by fluorescence microscopy, bone mineralization was visualized by calcein. The results show that AE and cyanidin‐3‐glucoside (C3G) increased osteoblast proliferation and reduced apoptosis in hFOB 1.19 osteoblasts induced by serum and glucose starvation. Gene expression data showed that C3G increased HDAC1 expression, increased BCL‐2 and reduced BAX mRNA and caspase 3/7 in apoptotic hFOB cells. AFE increased the proliferation of osteoblasts in dT osx/Sp7:mCherry medaka by enhancing the expression of osx/Sp7. Osterix (also known as transcription factor Sp7) is an osteoblast‐specific transcription factor which activates a repertoire of genes during differentiation of preosteoblasts into mature osteoblasts and osteocytes. AFE and C3G enhanced proliferation of hFOB cells, reduced hFOB apoptosis induced by serum and glucose starvation by via epigenetic regulation of HDAC1, and by increasing BCl‐2 and reducing the expression of BAX and caspase 3/7. The AFE enhanced osteoblast proliferation in osx/Sp7:mCherry medaka, and increased osx/sp7 expression.Support or Funding InformationNCCIH at NIHThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Therapeutic effects of 5‐HT2A receptor activation in a murine ovalbumin‐induced chronic asthma model
Authors: Flanagan Thomas Wayne 
Published: June 2021

Asthma is a chronic respiratory disease characterized by airway hyperresponsiveness, bronchial inflammation, and mucus overproduction. Most in vivo murine models of asthma utilize short‐term exposure of an aerosolized antigen to induce a T‐helper‐2 cell triggered immune response. While these models provide necessary insight into the pathophysiological mechanisms governing acute asthmatic attacks, they are limited in their ability to promote the airway structural remodeling and persistent airway hyperresponsiveness which characterize clinical asthma. Here we have generated a persistent model of allergic airways disease to evaluate the effects of serotonin 2A receptor activation on chronic asthma. We have previously demonstrated that intranasal delivery of the highly selective 5‐HT2A agonist (R)‐2,5‐dimethoxy‐4‐iodoamphetamine [(R)‐DOI] potently prevents mucus hyperproduction, airways inflammation, and pulmonary eosinophil recruitment in an acute model of allergic asthma. In this work prolonged exposure to aerosolized ovalbumin induces persistent airway hyperresponsiveness, goblet cell hyperplasia, and elevated collagen deposition with increased smooth muscle mass, all hallmarks of chronic asthma in humans. Inhalation of 1.0 mg/kg (R)‐DOI post allergen exposure reduces airway hyperresponsiveness and collagen deposition in mice with persistent allergic airways disease. Furthermore, 5‐HT2A receptor activation resets the inflammatory microenvironment repeated chronic allergen exposure induces in the whole lung and airways. These results highlight a unique role the 5‐HT2A receptor plays in immune cell regulation and further illustrates 5‐HT2A receptor agonism to be a powerful anti‐inflammatory therapy.Support or Funding InformationThis work was supported, in part, by research funds from the American Asthma Foundation Scholar Grant Award to C.N. and a sponsored research contract from The Eleusis Benefit Corporation.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Bifidobacterium breve and Lactobacillus rhamnosus Attenuate the Inflammatory Mediators Secretion in a Human Bronchial Epithelial (BEAS) Cells Culture Stimulated with Cigarette Extract
Authors: Aimbire Flavio , Miranda Munique , Sá Ana Karolina , Carvalho Jose Luis 
Published: June 2021

Abnormal inflammatory response in lung tissue is the main characteristic of chronic obstructive pulmonary disease (COPD), mainly triggered by exposure to cigarette smoke. COPD is one of the major causes of morbidity and mortality, with a great socioeconomic impact worldwide. Despite the importance of the disease and its impact, existing treatment still presents limitations and the presence of several side effects. Therefore, the search for alternative therapies is extremely relevant and the use of probiotics in the treatment of COPD has been highlighted, due to its immunomodulatory potential in chronic inflammatory diseases. Our study proposes to evaluate the role of Bifidobacterium breve (Bb) and Lactobacillus rhamnosus (Lr) probiotics on the in vitro inflammatory profile of cytokine released after stimulation of bronchial epithelial cells (BEAS) with cigarette smoke extract. We stimulated human derived BEAS culture with 2.5% cigarette extract, produced from the burning of 1 cigarette, whose smoke was incorporated into the culture medium (10 ml) through a vacuum pump (−11Kpa). After four hours, Lr or Bb was added to the BEAS culture. 24 hours after probiotic administration, the supernatant was collected to be measured by ELISA. The extract was sufficient to stimulate the cell lineage. Both Lr and Bb were efficient in reducing levels of IL‐6, IL‐1β, TNF‐α, CXCL1, CXCL5, CXCL8 and CXCL9, while increasing IL‐10 and TGF‐β levels. The probiotics proposed modulate the synthesis of pro‐inflammatory mediators, reducing the concentration of cytokines and chemokines secreted in the BEAS culture supernatant.Support or Funding InformationFAPESPThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Obesity impairs ADAM17‐mediated vascularization of pericardial adipose tissue in patients with coronary artery disease
Authors: Bagi Zsolt , Balogh Marta , Davila Alec , Tian Yanna , Patel Vijay , Czikora Istvan 
Published: June 2021

Expansion of pericardial adipose tissue (pAT) correlates with the severity of coronary artery disease in obese patients. Dysfunctional pAT is a significant source of pro‐inflammatory cytokine and adipokine production, but mechanisms that underlie pAT dysfunction remain poorly understood. During angiogenesis vascular endothelial growth factor (VEGF) activates A disintegrin and metalloproteinase ADAM17 that induce Notch ectodomain shedding to promote new vessel growth. We hypothesized that ADAM17‐mediated vascularization of expanded pAT is disproportional in obese patients. We obtained pAT from patients undergoing coronary artery bypass surgery (N=38, BMI, 17–43) and found that BMI was positively correlated with adipocyte size and increased collagen deposition in pAT. A higher BMI was associated with lower ADAM17 activity in pAT microvessels, which was associated with significantly reduced microvessel density and reduced capillary to adipocyte ratio in obese patients. In a three‐dimensional angiogenesis assay we determined VEGF‐stimulated growing of endothelial sprouts from pAT artery rings. We found that the number of endothelial sprouts is significantly reduced in obese patients when compared to that of pAT arteries from non‐obese subjects. We also found that genetic silencing of ADAM17 significantly reduced VEGF‐induced endothelial sprouting. Thus, we concluded that reduced ADAM17 activation results in impaired vascularization and hence dysfunctional pAT in obese patients with coronary artery disease.Support or Funding InformationAmerican Heart Association 17GRNT33680171, NIH F31 HL142183‐01 and NIH R01AG054651This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endothelial TRPV4 channel deletion promotes tumor growth and metastasis
Authors: Kanugula Anantha Koteswararao , Adapala Ravi K , Jamaiyar Anurag K , Lenkey Nina , Yin Liya , Chilian William M , Paruchuri Sailaja , Thodeti Charles K 
Published: June 2021

Ion channels have been implicated in tumor growth and metastasis and are considered as one of the promising therapeutic targets. Previously, we have shown that transient receptor potential vanilloid 4 (TRPV4) ion channel is a mechanosensor in endothelial cells (EC) which is downregulated in tumor endothelial cells (TEC). TEC exhibit high basal Rho activity, abnormal migration and angiogenesis in vitro. Mechanistically, we found that TRPV4 regulates EC migration and angiogenesis by integrating mechanical (Rho) and soluble (VEGF) signaling. Importantly, we found that tumor implantation in global TRPV4KO mice resulted in enhanced tumor growth, angiogenesis and lung metastasis compared to wild type mice suggesting that TRPV4 in endothelium negatively regulates tumor angiogenesis. However, the specific role of endothelial TRPV4 in tumor growth, angiogenesis and metastasis is not known. To determine this, we have generated endothelial specific TRPV4KO (TRPV4ECKO) mice by crossing TRPV4 lox/lox mice with Tie2‐Cre mice. By employing syngeneic Lewis lung carcinoma tumor model, we found that deletion of endothelial TRPV4 enhanced tumor growth in TRPV4ECKO mice compared to TRPV4 lox/lox mice. Multiphoton microscopic analysis of tumor sections infused with Alexa‐Fluor isolectin‐B4 revealed tortuous tumor vasculature in TRPV4ECKO mice compared to TRPV4 lox/lox mice. Further, we found that these vessels show reduced pericyte coverage with concomitant leakiness as evidenced by colocalization of CD31/α‐SMA and dextran leakage, respectively. Western blot analysis revealed increased tyrosine phosphorylation of VEGFR2 at Y1175 (p‐VEGFR2‐Y1175) in tumors from TRPV4ECKO mice compared to TRPV4 lox/lox. Finally, we found increased metastasis of tumor cells to the lung in TRPV4ECKO mice. Our results thus confirm that endothelial TRPV4 is a negative regulator of tumor angiogenesis and metastasis and identifies TRPV4 is a novel VEGF‐independent target for tumor angiogenesis and metastasis or vascular normalization.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Alteration of Calcium and Electrical Dynamics in Cerebrovascular Endothelium During Development of Alzheimer's disease
Authors: Hakim Md A. , Buchholz John N. , Behringer Erik J. 
Published: June 2021

Alzheimer's disease (AD) is a progressive neurodegenerative disease and the most common cause of dementia characterized by a decline in cognitive function among the elderly. Currently, ~5.7 million Americans are living with AD and this number will increase to ~14 million by 2050. Age‐related AD alters blood flow to the brain and is associated with cerebral hypoperfusion, due in part, to impaired vascular endothelial function. However, it is unknown whether AD pathology impacts the biophysical functions of key cerebral vascular G‐protein‐coupled receptors (GPCRs) and K+ channels (KCa2.3 or SKCa, KCa3.1 or IKCa) that govern blood flow via an electrical vasodilatory pathway known as endothelium‐derived hyperpolarization (EDH). Thus, we tested the hypothesis that cerebrovascular endothelial purinergic receptors and Ca2+‐activated K+ channels become functionally dysregulated during progression of AD. We used the triple mutation mouse model of AD (3xTgAD) to examine endothelium isolated from posterior cerebral arteries in young control (YC; 1–2 mo), amyloid‐β plaques (Aβ; 6–8 mo), and plaques + neurofibrillary tangles (Aβ+Tau; ≥ 12 mo); n ≥ 3 male & n ≥ 3 female mice/group. Intracellular calcium concentration ([Ca2+]i) and membrane potential (Vm) were simultaneously measured using Fura‐2 photometry and sharp electrodes (pH 7.4, 37°C). EDH was demonstrated by an increase in [Ca2+]i (ΔF340/F380 ≥ 0.3) concomitant with ΔVm ≥ −10 mV in response to the P2Y agonist ATP (100 μM, 3 min). Membrane hyperpolarization to ATP during Aβ+Tau vs. YC decreased (ΔVm, mV; YC: −20±4, Aβ: −17±2, Aβ+Tau: −13±3) while [Ca2+]i responses were maintained. In contrast, hyperpolarization increased during Aβ and Aβ+Tau (ΔVm, mV; YC: −29±3, Aβ: −36±2, Aβ+Tau: −38±3) during direct activation of SKCa/IKCa alone with SKA‐31 (10 μM, 5 min). Finally, Δ[Ca2+]i and ΔVm responses to hydrogen peroxide (H2O2; 200 μM, 20 min) in young control animals were greater by > 40% as compared to Aβ & Aβ+Tau. These data suggest coupling of endothelial SKCa/IKCa function with Ca2+ signaling is reduced and adaptation to oxidative stress occurs during the development of AD. Altogether, endothelial K+ channel function may be directly calibrated for optimal cerebral blood flow to maintain a healthy brain with aging while helping to prevent neurodegenerative disease.Support or Funding InformationThis research is supported by Loma Linda University School of Medicine new faculty start‐up funds and National Institutes of Health grants R00AG047198 & R56AG062169 (EJB).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of Human Endothelial Cell Function by Metformin and Canagliflozin
Authors: Durante William , Behnammanesh Ghazaleh , Durante Zane E. , Peyton Kelly J. 
Published: June 2021

Metformin is the drug of first choice in treating hyperglycemia in patients with type 2 diabetes mellitus. Its primary action is to inhibit hepatic glucose production and increase the sensitivity of peripheral tissues to insulin. Recently, sodium‐glucose cotransporter 2 inhibitors such as canagliflozin, which reduce blood glucose levels by blocking glucose reabsorption in the kidney, have also been approved for use in type 2 diabetes mellitus either alone or in combination with other drugs, including metformin. Intriguingly, both metformin and canagliflozin have been shown to improve cardiovascular outcomes in patients with type 2 diabetes mellitus. While reductions in body weight, adiposity, blood pressure, and arterial stiffness have been suggested to contribute to the cardiovascular benefit of these drugs, their effects on vascular cell function have not been fully considered. In the present study, we tested the hypothesis that metformin and canagliflozin directly effect human endothelial cell function. Treatment of human umbilical vein endothelial cells with metformin or canagliflozin inhibited their proliferation in a concentration‐dependent manner. The anti‐proliferative action of both drugs was associated with a significant reduction in DNA synthesis and the arrest of endothelial cells in the G0/G1 phase of the cell cycle. While pharmacologically relevant concentrations of canagliflozin (10μM) blocked endothelial cell proliferation, much higher supra‐pharmacological concentrations of metformin were needed to repress endothelial cell growth. In addition, both drugs inhibited the migration of endothelial cells and the differentiation of endothelial cells into tubes in a concentration‐dependent fashion. Furthermore, incubation of endothelial cells with a clinically attainable concentration of metformin (10μM) potentiated the ability of canagliflozin to block endothelial cell proliferation, but not endothelial cell migration or differentiation. In conclusion, metformin and canagliflozin inhibits human endothelial cell proliferation, migration, and tube formation. Moreover, metformin potentiates the anti‐proliferative action of canagliflozin, demonstrating a novel and clinically relevant interaction between these two anti‐diabetic agents on endothelial cell function. The ability of these drugs to modify endothelial cell function either individually, or in combination, may contribute to their cardiovascular actions in patients with type 2 diabetes mellitus.Support or Funding InformationAmerican Diabetes Association Grant #1‐17‐IBS‐290.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.