bluuebunny/crossref_metadata_2025_split huggingface dataset:
TAK1 Inhibitor 5Z‐7‐oxozeaenol Sensitizes Glioblastoma to Chemotherapy
Authors: Casili Giovanna , Campolo Michela , Lanza Marika , Filippone Alessia , Cuzzocrea Salvatore , Esposito Emanuela 
Published: June 2021

Glioblastoma (GBM) are the most common and aggressive primary brain tumors of the central nervous system. Despite relevant progress in conventional treatments, the prognosis of such tumors remains almost invariably dismal. NF‐κB activation is one of the resistance mechanisms for cancer cells to escape from chemotherapy‐induced cell‐death. TAK1 is an essential component in genotoxic stresses‐induced NF‐κB activation; however, the role of TAK1 in the development of chemoresistance in GBM remains unknown. The aim of the present study was to verify, in GBM human cell lines U138 and A172, if TAK1 inhibition significantly enhances the sensitivity of GBM cells to chemotherapy treatments. Also, to confirm the translationality of the study, GBM patients samples were used. Using GBM cell lines, U138 and A172, we found that TAK1 inhibitor 5Z‐7‐oxozeaenol significantly augmented the cytotoxic effects of temozolomide (TMZ). TAK1 inhibition also enhanced the inhibitory effect of TMZ on anchorage independent growth. Moreover, treatment of GBM cells with 5Z‐7‐oxozeaenol blocked TMZ‐ induced NF‐κB activation, reduced cytokines expression and enhanced TMZ‐induced apoptosis in both cell lines. Moreover, since MAPK are involved in the activation of NF‐κB, we investigated the MAPK role in GBM patients tissue, suggesting an effective role of MAPK in GBM modulation, highlighted by an up‐regulation of p‐p38 and pJNK in GBM patients samples. Together, our results provide a proof‐of‐concept that TAK1 inhibition significantly increases the sensitivity of GBM cells to chemotherapy‐induced cell‐death and can serve as an effective adjunct to current chemotherapeutic regimens for high‐risk diseases.Support or Funding InformationNo fundingThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A putative PPAR beta/delta agonist induces cell death in human cancer cell lines
Authors: Strom David Kelly , Li Dalon , Brinkman Hanna , Gleason Stephanie , Hershberger Sarah , Cumbay Medhane , LaFontaine Michael , Boncher Tracey , Hegwood Emma 
Published: June 2021

Compound 9 was specifically developed as an agonist for PPAR beta/delta. Binding studies showed it also bound PPAR gamma. Because of the recent identification of PPAR beta/delta involvement in a number of cancers, we tested whether compound 9 would synergize with known chemotherapeutic agents to increase apoptosis in a human cancer cell line that expresses the PPAR beta/delta receptor. These studies revealed that compound 9 was capable of inducing apoptosis on its own, and did not synergize with doxorubicin in inducing apoptosis. We have undertaken a series of studies to determine if the apoptosis that compound 9 induces is being mediated through any of the PPAR receptors. We have also examined compound 9 ability to signal through the PPAR receptors in a variety of cell lines.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Downregulation of Guanylate Cyclase Enzyme in Human Asthma model to Investigate NO‐sGc‐ cGMP as a Therapeutic Pathway in Asthma
Authors: London Sabina , Koziol‐White Cynthia , Guo Changjiang , Panettieri Reynold , Gow Andrew 
Published: June 2021

Asthma is characterized by airway smooth muscle hypersensitivity. Common treatments include β‐adrenergic agonist therapy, such as budesonide, to induce airway smooth muscle relaxation via increased cAMP. However, 70% of patients develop resistance to such therapies. Nitric oxide is a locally produced smooth muscle relaxant that operates via stimulation of soluble guanylate cyclase (sGc) to increase cGMP production. In this study, the NO‐sGc‐cGMP pathway was investigated as an alternate pharmacologic pathway for asthma therapy. Previous studies have found that asthma patients are insensitive to NO signaling and have used sGc drugs to directly stimulate the enzyme. The objective of this study was to examine the expression of the two sGc subunits, α and β, within smooth muscle cells isolated from asthma patients in order to understand how NO‐sGC‐cGMP signaling may be disrupted. Further, we examined how budesonide treatment altered sGc expression. RNA sequencing data showed increased mRNA for both sGc subunits in asthmatic patients and that budesonide downregulated the gene expression of both subunits. We hypothesized that this increased gene expression was in part due to a lack of cGMP signaling due to decreased protein levels that is further impaired with steroids such as budesonide. To examine sGc, we measured the protein levels of both the α and β subunits of sGC by western blot in cultured smooth muscle cells. Fatal asthma and non‐asthma donor derived primary airway smooth muscle cells were cultured with and 24 hours of budesonide treatment. Statistical analysis of western blotting was performed by ANOVA. Densitometry of sGC western blots indicated a 2‐fold downregulation of the α (p<0.0001) and 1.5 fold downregulation of the β (p<0.05) subunits in asthmatic compared to normal patients. No significant changes were observed with normal or asthmatic patients with budesonide treatment (p>0.05), indicating no effect of steroids on the NO‐sGc‐cGMP pathway. A downregulation of sGc protein levels in asthmatic patients would lower cGMP production, which could explain the lack of NO responsiveness in these patients and would contribute to increased airway constriction in fatal asthma. These results indicate the NO‐sGc‐cGMP pathway as a potential new therapeutic pathway in asthma, and future work will examine cGMP levels.Support or Funding InformationSupported by R25ES020721 and ASPET SURF Intern ProgramThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cerebrovascular Compliance is Affected by Posture
Authors: Moir M. Erin , Zamir Mair , Klassen Stephen A. , Balestrini Christopher S. , Shoemaker J. Kevin 
Published: June 2021

The mechanical properties of the arterial vascular wall influence the oscillatory component of flow through a vascular bed. Windkessel modeling has often been used to non‐invasively estimate these mechanical properties. While the systemic and coronary circulation have received considerable attention, the cerebral circulation represents a unique challenge because the oscillatory component of cerebral blood flow occurs within a pressurized and rigid skull. The present investigation evaluated the compliance of the cerebral vascular bed in comparison to that of the forearm vascular bed, a bed without significant extravascular pressure. Six young and healthy adults (24 ± 3 years, 5 females) were instrumented with a 3‐lead electrocardiogram, a finger photoplethysmograph (Finapres Medical Systems), and transcranial Doppler ultrasound (Multigon Industries) for continuous measures of heart rate, arterial blood pressure from the brachial artery, and cerebral blood flow velocity from the middle cerebral artery, respectively. Brachial artery blood flow velocity was also measured (duplex Doppler ultrasound; GE Healthcare). Following stabilization of hemodynamic variables, one to two minutes of data were collected in the supine posture. Measures were repeated after participants transitioned to a sitting posture, following re‐stabilization of variables. A modified Windkessel model was used to calculate the values of vascular compliance, as well as viscoelasticity and inertance effects, that are required to match the measured flow waveform to that predicted from the measured pressure waveform. Estimates of compliance were calculated for the forearm and cerebral vascular beds. In the supine posture, compliance of the cerebral vascular bed was lower than that of the forearm vascular bed by more than one order of magnitude (0.0006 ± 0.0005 vs. 0.002 ± 0.0007 mL/mmHg; P≤0.001; d=2.6). However, when seated, cerebrovascular compliance and forearm vascular compliance were not significantly different (0.002 ± 0.001 vs. 0.003 ± 0.002 mL/mmHg; P=0.10; d=1.0). The compliance in the brain increased considerably from the supine to sitting posture (P=0.03; d=0.91) but no significant difference was observed in the forearm (P=0.13; d=−0.65). The results suggest that intracranial pressure may prevent the cerebral vessels from exercising their compliance in supine. Further, as cerebrovascular compliance is increased with sitting, we speculate that posture‐induced reductions in cerebral venous volume affect cerebrovascular mechanics.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Junctional Pericytes Serve as Directional Control Elements in K+‐mediated Functional Hyperemia
Authors: Gonzales Albert Louis , Nelson Mark 
Published: June 2021

K+‐dependent activation of capillary endothelial Kir channels initiates a retrograde hyperpolarizing signal that causes upstream arteriolar dilation and increased blood flow into the capillary network. These findings cast capillary networks as dynamic K+‐sensing webs that respond to regional brain activity by converting neuronal activity, detected as changes in the extracellular ionic environment, into rapid increases in capillary blood flow. Pericytes cover a significant portion of the vast capillary network within the brain, with a majority of such cells located at capillary junctions. Because they are coupled by gap junctions, pericytes can receive electrical signals from the underlying endothelial cells. In addition, pericytes closest to the terminal arteriole possess multiple capillary‐wrapping projections that are dynamically and independently contractile. Accordingly, we hypothesized that different projections of a contractile pericyte are capable of receiving and isolating branch‐specific stimulation (hyperpolarization), leading to relaxation and changes in the fractional distribution of red blood cells (RBCs) to favor blood flow in the direction of the stimulus. Using in vivo multiphoton microscopy, we determined the relative distribution of RBCs at capillary junctions in response to the local delivery of 15 mM K+ to capillary segments downstream of one branch. K+‐dependent activation of capillary endothelial cells led to an asymmetric shift in RBC flux, such that flux robustly increased in the stimulated branch and decreased or did not change in the other branch. By contrast, picospritzing 15 mM K+ onto a capillary segment downstream of one branch of a distal, non‐contractile junctional pericyte caused a symmetrical increase in blood flow in both daughter branches. These results are consistent with the idea that pericytes deeper in the capillary network lack the ability to dynamically redistribute blood, and imply that the observed increase in blood flow occurred upstream of the distal junctional pericyte. Collectively, our findings indicate that contractile pericytes are capable of receiving propagating K+‐induced hyperpolarizing signals through the capillary network and dynamically manipulating regional blood flow through reallocation and redistribution of RBCs.Support or Funding InformationTotman Medical Research Trust, Foundation Leducq, Horizon 2020, NIH R01HL131181, R01HL‐121706, R37DK053832, and K01HL138215.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inducible Deletion of Endothelial Hba1 Significantly Reduces Exercise Fitness in Mice
Authors: Keller Alexander S. , Brooks Steven , Islam Aditi , Keller TC Stevenson , Best Angela K. , Cortese‐Krott Miriam K. , Yan Zhen , Ackerman Hans , Isakson Brant E. 
Published: June 2021

Hemoglobin (Hb) alpha is uniquely expressed in the endothelial cells of resistance arteries. The same protein is generated by both Hba1 and Hba2 genes. We have shown that siRNA knockdown of endothelial Hba1 decreases Hb alpha protein, and increases NO signaling in the vascular wall. Furthermore, pharmacologic inhibition of the interaction between endothelial Hb alpha and endothelial nitric oxide synthase reduces blood pressure. To further study the role of endothelial Hb alpha, we have produced a novel genetically modified mouse in which exons 2 and 3 of the Hba1 gene are flanked by loxP sites (Hba1fl/fl). We have crossed the Hba1fl/fl mice with a Cdh5‐Cre/ERT2+ mice to produce a tamoxifen‐inducible genetic model in which Hba1 can be temporally deleted from endothelial cells (EC Hba1fl/fl). We compared EC Hba1fl/fl with Hba1−/− (global deletion of Hba1) and found both mice exhibit reduced expression of Hba protein in skeletal muscle endothelial arteries and arterioles. However, while Hba1−/− mice showed significant decreases in several red blood cell parameters including hemoglobin concentration, hematocrit, erythrocyte count, red cell volume, and red cell hemoglobin, EC Hba1fl/fl red blood cell parameters were all unchanged. In order to test roles of endothelial alpha globin in hypoxic blood flow control, we used a treadmill running test to measure acute endurance capacity, as well as long‐term measurement of voluntary running volume in running wheel‐equipped cages. We found that in both experimental conditions, male EC Hba1fl/fl mice had a significant reduction in exercise capacity, mirroring the exercise phenotype of Hba1−/− mice. We conclude that endothelial Hba1 is an important regulator of vascular function and skeletal muscle performance.Support or Funding InformationAmerican Heart Predoctoral Fellowship #16PRE30240007, NIH R01 HL088554This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mitochondrial‐Targeted Antioxidant (MitoQ) Improves Vascular Function in Healthy Late Middle‐Aged and Older Adults
Authors: Rossman Matthew J. , Santos‐Parker Jessica R. , Steward Chelsea A.C. , Bispham Nina Z. , Cuevas Lauren M. , Rosenberg Hannah L. , Woodward Kayla A. , Chonchol Michel , Gioscia‐Ryan Rachel A. , Murphy Michael P. , Seals Douglas R. 
Published: June 2021

Excess reactive oxygen species production by mitochondria (mtROS) is a key mechanism of vascular dysfunction with aging. Our laboratory has shown that supplementation with the mitochondrial‐targeted antioxidant MitoQ improves vascular endothelial function by reducing mtROS and ameliorates arterial stiffening in old mice, but the effects in humans are unknown. Here we sought to translate our preclinical findings to humans, while also determining the safety and efficacy of MitoQ. Twenty healthy late middle‐aged and older adults (60–79 years) with impaired endothelial function (baseline brachial artery flow‐mediated dilation [FMD] <6%) underwent six weeks of oral supplementation with MitoQ (20 mg/day) or placebo in a randomized, placebo‐controlled, double‐blind, crossover design study. MitoQ was well tolerated, and no treatment‐related serious adverse events occurred. Plasma MitoQ concentrations were higher after the treatment vs. placebo period (P<0.05). Brachial artery FMD was 42% higher after MitoQ vs. placebo (P<0.05); this improvement was associated with complete amelioration of tonic mtROS‐related suppression of endothelial function (assessed as the increase in FMD with acute 160 mg dose of MitoQ, n=9, P<0.05). Aortic stiffness (carotid‐femoral pulse wave velocity [PWV]) was lower after MitoQ vs. placebo (P<0.05) in participants with elevated baseline levels (carotid‐femoral PWV >7.60 m/s, n=11). Plasma oxidized low‐density lipoprotein, a marker of oxidative stress, also was lower after MitoQ vs. placebo (P<0.05), whereas participant characteristics, endothelium‐independent dilation (sublingual nitroglycerin) and circulating markers of inflammation were not different (all P>0.1). These findings in humans support earlier preclinical observations and suggest that MitoQ and other therapeutic strategies targeting mtROS may hold promise for treating age‐related vascular dysfunction.Support or Funding InformationClinicaltrials.gov Identifier: NCT02597023Funding: Supported by NIH R21 AG049451, T32 AG000279, F32 AG053009, Colorado CTSA UL1 TR001082 and an industry contract with MitoQ Limited.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
H2O2 Regulates Arachidonic Acid‐induced TRPV4‐mediated Vasodilation in Human Coronary Arterioles
Authors: Zinkevich Natalya S. , Gutterman David D. , Zhang David X. 
Published: June 2021

Previous studies have demonstrated that arachidonic acid (AA) induces potent dilation of human coronary arterioles (HCAs) through the activation of endothelial TRPV4 channels followed by endothelial hyperpolarization or release of vasoactive substances. In addition, TRPV4‐mediated vasodilatory response to flow is augmented in HCAs from subjects with coronary artery disease (CAD), a condition associated with increased H2O2 production. However, the role of H2O2 in regulation of TRPV4 function remains to be elucidated. Therefore, we designed this study to examine the effect of endogenous H2O2 on TRPV4 function. Coronary arterioles (~150 μm ID), isolated from atrial tissues from patients with CAD, were cannulated and pressurized at 60 mmHg for videomicroscopic measurement of vessel diameters. In arterioles preconstricted with endothelin‐1, AA (10−11 to 10−6 M) elicited a concentration‐dependent dilation (maximal dilation: 84±5%, n=5), and this dilation was largely blocked by HC‐067047 (10−6 M), a TRPV4 antagonist (17±4%, n=5). Intraluminal incubation of HCAs with catalase, a H2O2 –scavenging enzyme (500 units/mL), also markedly reduced the dilation to AA (57±5% vs. 85±5% of control, n=8). The EET antagonist 14,15‐EEZE (10−5 M) failed to attenuate AA‐induced dilation (64±6% vs. 70±7% of control, n=5), suggesting that P450 metabolites of AA such as EETs do not significantly contribute to endothelial TRPV4‐mediated response. Together, these data provide evidence that H2O2 regulates AA‐induced TRPV4‐mediated dilation in HCAs.Support or Funding InformationThis work was supported by NIH R01‐HL096647 (to David X Zhang)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of NRF2 Activation on Endothelial Function, Microvessel Density, and Gene Expression in Rats fed High Salt Diet
Authors: Uche Nnamdi , Kozak Kaleigh , Fink Katie , McCord Joe , Lombard Julian H. 
Published: June 2021

BACKGROUNDHigh salt (HS) diet promotes vascular oxidative stress and endothelial dysfunction in animal models and humans. Studies in animals indicate that salt‐induced suppression of antioxidant defense mechanisms plays a crucial role in endothelial dysfunction, microvascular rarefaction and vascular oxidative stress. We hypothesized that activation of the master antioxidant transcription factor nuclear factor (erythroid‐derived 2)‐like‐2 (NRF2) with the dietary supplement Protandim protects against salt‐induced vascular dysfunction and microvascular rarefaction by restoring redox homeostasis.OBJECTIVETo determine whether activation of Nrf2 via dietary supplementation with Protandim will prevent salt‐induced endothelial dysfunction, vascular oxidative stress, and microvascular rarefaction via effects on antioxidant enzyme expression.METHODSAge‐matched, male Sprague‐Dawley rats (12–15 weeks old) were fed a high salt (HS; 4.0% NaCl) diet ± ~60 mg/kg/day Protandim supplement for 2 weeks. mRNA from harvested liver samples was used for real time PCR evaluation of the effects of Nrf2 upregulation on a range of genes related to oxidative stress, as reported in the literature. Changes in protein expression were evaluated via Western blotting.RESULTSProtandim treatment restored endothelium‐dependent dilation to acetylcholine in middle cerebral arteries, reduced mitochondrial ROS levels (Mito‐SOX) in basilar arteries, and increased microvessel density in the cremaster muscle of HS‐fed rats. Protandim supplementation increased mRNA levels of thioredoxin reductase 1 (n=6, p=0.008), reduced mRNA expression of the pro‐oxidative transcription factor Kruppel‐like factor 9 (Klf9) (n=6, p < 0.001) and tended to increase the expression of mRNA for glutathione reductase (GSR) and catalase in liver samples compared to untreated rats fed HS diet alone. Other indicators were not significantly changed by Protandim treatment, either by qPCR or Western blot analysis.CONCLUSIONSTreatment of HS‐fed rats with the Nrf2 activator Protandim ameliorates salt‐induced endothelial dysfunction, vascular oxidative stress, and microvascular rarefaction via complex effects on enzymes and proteins affecting redox state.Support or Funding InformationNIH #R01‐HL128242 and # R21‐OD018309‐JHLThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cx43 Response of Endothelial Cells to DHA is Not Mediated by the FFAR4
Authors: Sundblad Morgan , Kurjiaka David 
Published: June 2021

The health of endothelial cells is important in the formation and progression of atherosclerotic plaques. These cells respond to factors circulating in the blood including free fatty acids (FFA). These FFA are linked to altered risk of cardiovascular diseases (CVD): saturated FFA increase whereas Omega3 polyunsaturated FFA decrease CVD risk. FFA structure may influence plaque growth by affecting the localized inflammatory response. Omega3 FFA bind to the FFA receptor 4 (FFAR4). As the health of the overlying endothelium is important in plaque formation, we were interested in the impact of Omega3 FFA docosahexaenoic acid (DHA) on endothelial cell (EC) inflammatory responses. Connexin 43 expression was evaluated as a marker of inflammation (Cx43 expression increases with inflammation). bEnd.3 cultured EC were treated with 30 uM DHA and protein isolated at 1.5, 3, 6, 12, 24, and 48 hrs along with a control at the same time. Whole cell protein was separated in a Western blot (equal loading), transferred to a membrane and probed for Cx43. EC response to DHA were time dependent: while EC Cx43 expression increased at 6 hrs, its expression had decreased by 24 hrs. At 24 hrs, blocking the FFAR4 receptor with AH‐7614 (5 uM) did not alter the DHA response. Interestingly, blocking the FFAR4 receptor decreased Cx43 expression at 24 hrs. Thus, the DHA induced decrease in Cx43 expression did not involve the FFAR4 receptor. The identity of the receptor mediating the Cx43 response to DHA may be the FFAR1 or could involves its conversion to prostaglandins.Support or Funding InformationGVSU Office of Undergraduate Research Student Summer ScholarsThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.