bluuebunny/crossref_metadata_2025_split huggingface dataset:
Phenotypic characteristics of pulmonary supernumerary arterial endothelium may be involved in the formation of plexiform lesions (1089.21)
Authors: Oshima Kaori , Matsumoto Yuri , O'Neill Kealan , McMurtry Ivan , Stevens Troy , Oka Masahiko 
Published: June 2021

The plexiform lesion (PL), which comprises proliferative endothelial marker (EC)‐positive channel cells and chromatin‐rich core cells, is the histological hallmark of severe pulmonary arterial hypertension (PAH). PLs preferentially form in supernumerary arteries (SAs) in SU5416/hypoxia/normoxia‐exposed PAH rats, which supports clinical observations. However, the underlying pathological mechanisms of this observation are unknown. SAs are unique small pulmonary branches with specific anatomical and functional properties, such as the phenotype of the ECs, which may contribute to the formation of PLs. The EC phenotype in PLs seems to be shared with pulmonary microvascular ECs (PMVECs), which are highly proliferative. Therefore, we tested the hypothesis that the ECs in SAs share the same phenotype as PMVECs that predisposes to the development of PLs.A lectin protein from Griffonia simplicifolia (GS), which selectively interacts with PMVECs was perfused through the pulmonary vasculature of control and PAH rats. Lung slices were prepared and examined by confocal fluorescence microscopy.In normal rats, ECs in SAs interacted with GS. However, ECs in SAs and occlusive vascular lesions in PAH rats had no apparent interaction with GS.The PMVEC‐like phenotype of ECs in SAs may contribute to the development of PLs, and there may be a phenotypic transformation of these cells during the progression of PAH.Grant Funding Source: Supported by NIH HL66299 and HL60024 to T.S.
Preservation of Ca2+ spark activity during oxidative stress in pulmonary arterial myocytes of fetal sheep (1089.5)
Authors: Suarez Gema , Glasgow Shane , Romero Monica , Osman Noah , Puglisi Jose , Bers Donald , Izu Leighton , Longo Lawerence , Wilson Sean 
Published: June 2021

Antenatal chronic hypoxia can lead to pulmonary hypertension in newborns or more severe vascular remodeling and dangerous elevations in pressure. We have previously shown that antenatal chronic hypoxia deregulates Ca2+ spark activity, which are important to pulmonary vasodilation at birth. Chronic hypoxia can also increase oxidative stress, which can impair Ca2+ spark activity. In this research, we examined the impact of oxidative stress on Ca2+ spark activity, which other laboratories have shown to increase Ca2+ spark activity. We exposed pulmonary arteries from normoxic fetal sheep to reactive oxygen species (ROS) to mimic oxidative stress such as might occur with acute hypoxic stress by treating them with tert‐butyl H2O2 and reduced ROS with the antioxidant N‐Acetyl‐L‐Cysteine (NAC). Using customized analysis software, roughly 7 % of control cells had Ca2+ sparks and the frequency was 0.07 sparks/100 μm/sec. Spark activity was maintained in the presence of tert‐butyl H2O2 and NAC. The spatial and temporal aspects to the Ca2+ sparks were then determined. Spark amplitudes were unaffected as was the width. However, Ca2+ spark decay was increased slightly by tert‐butyl H2O2. Overall, oxidative stress has only a mild influence on Ca2+ spark activity on fetal pulmonary arteries from normoxic sheep.Grant Funding Source: Supported by NSF MRI 923559, NIH HD69746, P01HD31226, R01HD3807, LLUSOM, APS Frontiers in Physiology
cGMP amplification of pulmonary arterial myocyte Ca2+ waves is preferentially impaired in high altitude‐induced hypoxic fetal sheep (1089.7)
Authors: Paez Ricardo , Romero Monica , Blood Quintin , Osman Noah , Manjunath Chetas , Glasgow Shane , Francis Michael , Taylor Mark , Longo Lawerence , Wilson Sean 
Published: June 2021

High altitude is a risk factor in the development of pulmonary hypertension (PH). Treatment often includes the use of phosphodiesterase (PDE) inhibitors of PDE3 (milrinone, cAMP) and PDE5 (sildenafil, cGMP), which increase cyclic nucleotides and promotes vasodilation in pulmonary arterial (PA) myocytes. With respect to the current studies, c‐AMP and c‐GMP enhance ryanodine receptor (RyR) and Sarco/Endoplasmic Reticulum Ca2+ ‐ATPase (SERCA) pump activity in myocytes, which regulate intracellular Ca2+. To test whether long term hypoxia (LTH) impairs, and maturity enhances the effects of c‐AMP and c‐GMP on myocyte Ca2+ waves, we selected 3‐isobutyl‐1‐methyl xanthine (IBMX), a PDE inhibitor, as well as 8‐Br‐c‐AMP and 8‐Br‐c‐GMP. We measured cytosolic Ca2+ in PA myocytes of fetal and adult sheep that lived at low (~353 m) or high altitude (~3801 m) via confocal imaging of fluo‐4. Maturation increased while IBMX, 8‐Br‐c‐AMP, and 8‐Br‐c‐GMP each failed to alter the percentage of responsive cells independent of altitude. LTH fetuses had reduced wave amplitude and area under the curve (AUC), but wave kinetics were unaltered. The AUC was maintained in LTH adults, but waves were faster and larger. c‐GMP increased AUC by lengthening waves in all groups except for hypoxic fetuses. Alterations of Ca2+ regulatory pathways including SERCAs and RyRs could contribute to LTH impairment in Ca2+ waves and c‐GMP amplified signals.Grant Funding Source: Supported by NSF MRI 0923559, NIH HD‐069746, P01HD031226, , R01HD003807, 5P20 MD‐006988,LLUSOM
Disruption of membrane caveolae limits pulmonary vasoconstriction induced by agonists in pulmonary hypertension rats (1089.8)
Authors: Jiao Hai‐Xia , Mu Yun‐Ping , Wang Rui‐Xing , Lin Da‐Cen , Sham James , Lin Mo‐Jun 
Published: June 2021

A rise in cytosolic Ca2+ in pulmonary artery smooth muscle cells (PASMCs) is the major trigger for pulmonary vasoconstriction. Previous studies of pulmonary hypertension (PH) show that canonical transient receptor potential (TRPC) is an important constituent of store‐operated Ca2+ channel (SOCC); TRPC genes are upregulated and store‐operated Ca2+ entry (SOCE) is augmented in PASMCs of monocrotaline (MCT)‐treated and chronic hypoxic (CH) rats. Since TRPC and caveolin may functionally colocalize in caveolae in PASMCs and coordinate to regulate pulmonary vascular tone, we sought to examine the effects of caveolae disruption on agonist‐induced contraction of isolated pulmonary arteries (PAs) of CH‐ and MCT‐induced PH rats. Rats developed severe PH and right ventricular hypertrophy within three weeks after CH‐(10% O2) exposure or single intraperitoneal injection of MCT at a dose of 60 mg/kg. The tension of PA rings induced by various agonists were compared with and without the caveolae‐disrupting agent methyl‐beta‐cyclodextrin (MCD) or cholesterol repletion with a cholesterol:MCD mixture (1:5). The MCD‐pretreatment attenuated PA contraction induced by cyclopiazonic acid (CPA), endothelin‐1(ET‐1) and phenylephrine (PE), but had no effect on KCl‐induced vasoconstriction in control groups. The cholesterol:MCD mixture rescued PA‐contraction induced by CPA, ET‐1 and PE. Compared with the control group, MCD‐pretreatment resulted in further inhibition of the CPA‐induced contraction of PAs of both CH‐ and MCT‐treated rats, as well as the PE‐induced contraction of PAs of MCT‐treated rats, but had no effect on KCl‐induced contraction in the CH‐ and MCT‐ groups. These results suggest that the activity of voltage‐dependent Ca2+ channels does not depend on the integrity of associated with caveolae, but the SOCC may be closely associated with the cholesterol‐enriched membrane lipid domains of PAs. Moreover, the enhanced SOCE of PAs caveolae in PH rats.Grant Funding Source: NSFC31171104 and FJMU09ZD010
Role of mitochondrial KATP channels in PKCβ‐dependent constriction of small pulmonary arteries (1090.10)
Authors: Snow Jessica , Norton Charles , Sands Michelle , Jernigan Nikki , Gonzalez Bosc Laura , Walker Benjimen , Resta Thomas 
Published: June 2021

PKCβ mediates vasoconstriction through a unique signaling pathway in the pulmonary circulation that involves mitochondrial reactive oxygen species (mitoROS). We hypothesized that PKCβ elicits these responses through activation of vascular smooth muscle mitochondrial KATP channels (mitoKATP). To test this hypothesis, we examined constriction in endothelium‐disrupted, pressurized rat pulmonary arteries (~150 μm diameter) and mitoROS production (by MitoSOX fluorescence) in cultured rat and human pulmonary arterial smooth muscle cells (PASMC) in response to the PKC agonist phorbol 12‐myristate 13‐acetate (PMA). PMA caused vasoconstriction and mitoROS production that were significantly attenuated (p<0.05) by both the PKCβ inhibitor LY‐333,531 (10 nM) and PKCβ siRNA. Consistent with our hypothesis, the specific mitoKATP channel antagonist 5‐hydroxydecanoate (5‐HD; 500 μM) inhibited vasoconstriction and mitoROS production to both PMA and the mitoKATP activator diazoxide (10‐5‐ 10‐3 M). Consistent with a role for mitoKATP in PKCβ‐dependent mitoROS generation, PMA caused mitochondrial membrane potential depolarization (detected by tetramethylrhodamine methyl ester fluorescence), which was prevented by 5‐HD (p<0.05 vs. vehicle) in both pulmonary arteries and human PASMC. We conclude that PKCβ enhances mitoROS production via mitoKATP activation in PASMC to mediate vasoconstriction.Grant Funding Source: Supported by NIH R01 HL088192 and American Heart Association grants 1POST5260026 and 13GRNT14
Interleukin‐33 in pulmonary arterial hypertension: a role in disease pathogenesis? (1090.4)
Authors: Aroonsakool Nakon , Titone Dominic , Li Jinghong , Dumouchel Justin , Lombardi Sandra , Kim Nick , Poch David , Bigby Timothy , Murray Fiona 
Published: June 2021

Pulmonary arterial hypertension (PAH) is characterized by increased pulmonary vascular resistance that leads to right ventricular failure. PAH can be a primary disease (idiopathic, IPAH) or secondary to conditions such as connective tissue disease. Interleukin‐33 (IL‐33), which is the ligand for the ST2 receptor, is a cytokine that acts as an alarmin. Since PAH is associated with the infiltration of inflammatory cells and increased cytokines, we aimed to investigate the role of IL‐33 in PAH. Using an IL‐33 ELISA we found that IL‐33 is increased in the serum of PAH patients, in particular IPAH [1798 +/‐846 pg/ml (n=6) vs. 24 +/‐ 13 pg/ml (n=6), p<0.01], compared to control subjects. We found that the mRNA expression of IL‐33 is increased ~8‐fold in peripheral blood mononuclear cells isolated from PAH‐patients and PAH‐animal models compared to control; Il‐33 is up‐regulated in the lung of animal models of PAH. Injury of pulmonary artery endothelial cells (PAEC) released IL‐33, thus implying that the increased circulating IL‐33 with PAH may occur if the membrane integrity of PAEC is compromised. Administration of IL‐33 (500 ng/day intracheally for four days) induced pulmonary arterial remodeling in WT and IL‐33 KO mice, but not ST2 KO mice. Our data demonstrate that IL‐33 is increased in PAH, which via ST2‐dependent signaling, contributes to remodeling of the PA and hence the pathophysiology of the disease.Grant Funding Source: Supported by NIH
Circulating microparticles from pulmonary arterial hypertension stimulate ICAM‐1 and E‐selectin expression in pulmonary artery endothelial cells (1090.6)
Authors: Hargett Leslie , O'Neill Kealan , Yarbrough Thomas , Bauer Natalie 
Published: June 2021

Circulating Microparticles from Pulmonary Arterial Hypertension stimulate ICAM‐1 and E‐selectin expression in Pulmonary Artery Endothelial CellsHargett L1, O’Neill K, Yarbrough T1, and Bauer N1,2. the Center for Lung Biology1 and Department of Pharmacology 2, University of South Alabama, College of Medicine, Mobile AL 36688.Microparticles (MPs) are submicron vesicles released by various cells that serve as mediators of inflammation. MPs harbor adhesion molecules and can stimulate the expression n of adhesion molecules in target cells. MPs are increased in pulmonary arterial hypertension (PAH). The hallmark PAH arterial vasculopathy involves inflammatory cell infiltration. Rho kinase (ROCK) is implicated in the expression of adhesion molecules in the pulmonary endothelium. We propose that MPs, generated as a result of PAH, activate ROCK and increasing the expression of adhesion molecules in pulmonary endothelium. MPs were isolated from the blood of normal and PAH rats (Su/Nx/Hx model). Pulmonary artery endothelial cells (PAECs) were treated with media, MPs from normal rats, or MPs from PAH rats. ICAM‐1 and E‐selectin, as assessed by western and flow cytometry, were increased in the PAECs. ROCK activity, as indicated by phosphorylated MYPT1696 western, was increased in PAH‐MP treated PAECs. Our results indicate that MPs from 8 week PAH rats increase the expression of ICAM‐1 and E‐selectin selectively in PAECs 6 hours after treatment. Also, preliminary data show increased phosphorylated MYPT1 in response to MPs from 8 week PAH rats. In conclusion, MPs from PAH rats increase adhesion molecule expression and may enhance ROCK activity in PAECs in vitro. As our studies progress, an inflammatory role for MPs in PAH could be revealed.This research is funded by AHA 11 (SDG7390037) Grant and NHLBI T32 (HL076125) Training Grant.
Repetitive acute intermittent hypoxia downregulates inflammatory gene expression in the rat ventral cervical spinal cord (1091.7)
Authors: Watters Jyoti , Roopra Avtar , Bowen Kellie , Vinit Stephane , Macfarlane Peter , Mitchell Gordon 
Published: June 2021

Intermittent hypoxia exerts pathologic and beneficial effects in the CNS, depending on the dose and duration. Whereas hundreds of hypoxic episodes per day mimic sleep apnea and promote neuroinflammation, “low dose” protocols, such as repetitive acute intermittent hypoxia (rAIH; 10 episodes of 10.5% O2 per day, 3 days/week for 4 weeks), upregulate growth/trophic factors and promote spinal motor plasticity. Because rAIH increases protein levels of key molecules associated with spinal respiratory motor plasticity, we hypothesized that the mRNAs for these and other molecules contributing to neuronal plasticity would be similarly increased. Nimblegen neuronal gene microarrays were used to profile transcript levels in the spinal ventral horn (C4‐C6) of male Lewis rats exposed to normoxia or rAIH (n=3 arrays/condition). Surprisingly, there was little evidence of increased mRNA levels of pro‐plasticity genes, and these results were confirmed with qRT‐PCR for specific genes of interest. Instead, there were significant decreases in inflammatory genes including cyclooxygenase‐2, tumor necrosis factor family members, chemokines and interleukins 1β and 6. To explore the cause of these changes, we used whole genome Vista analyses to identify the transcription factor binding sites over‐represented in the upstream 5kb promoter regions of genes affected by rAIH. Genes with binding sites for nuclear factor kappa B (NF‐kB) were the most over‐represented (p<0.005), suggesting that rAIH may decrease NF‐kB transcriptional activity. Thus, in contrast to chronic intermittent hypoxia, rAIH may be anti‐inflammatory, depleting spinal NF‐kB target genes.Grant Funding Source: Supported by NIH HL111598, HL080209, HL69064 and NS065067
Ventilatory sensitivity to CO2 is altered by external breathing resistance (1092.10)
Authors: Shykoff Barbara , Warkander Dan 
Published: June 2021

CO2 retention by divers underwater or by subjects exercising while breathing on a resistive circuit is uncorrelated with resting ventilatory sensitivity to CO2. We hypothesized that work of breathing might modulate sensitivity. Read’s rebreathing method was used to measure resting ventilatory sensitivity to CO2 with no added resistance, inspiratory, expiratory, or both resistances. External resistors were two plugs with holes 0.56”, measured to cause work of breathing per tidal volume of 1 kPa with minute ventilation 100 L/min. Ventilation was measured with a turbine flow meter. CO2 in the mouthpiece was monitored using a sector mass spectrometer. Preliminary data are available from 7 healthy Navy divers. Sensitivities are given in (L/min)/Torr for the initial increase in CO2; some subjects showed increased sensitivity once CO2 climbed. Median values were 1.9 without added resistance, 3.9 with inspiratory resistance, 2.3 with expiratory resistance, and 3.2 with both resistances. However, minima were 0.5 without resistance, 0.12 with inspiratory resistance, and negative with expiratory or both resistances. For all conditions, some subjects showed normal sensitivities only after CO2 had climbed above 52 to 60 Torr, and some showed no ventilatory response. In this small sample, resting ventilatory sensitivity to CO2 changed unpredictably with external resistance.Grant Funding Source: Supported by Naval Sea Systems Command Deep Submergence Biomedical Program
The thyrotropin‐releasing hormone analog Taltirelin augments the hypercapnic ventilatory response in CO2‐insensitive Brown Norway but not Sprague Dawley rats (1092.6)
Authors: Hodges Matthew , Novotny Tyler , Echert Ashley , Puissant Madeleine , Mouradian Gary 
Published: June 2021

Thyrotropin‐releasing hormone (TRH) is an excitatory neuropeptide that may modulate the hypercapnic ventilatory response (HCVR). We have found reduced expression of TRH in the medullary raphe in Brown Norway (BN) compared to Sprague Dawley (SD) rats. Herein we tested the hypothesis that the reduced TRH in the BN rat contributes to their blunted HCVR by measuring ventilation (VE), arterial blood gases, and rectal temperature (TR) in BN and SD rats during exposures to room air (RA) or hypercapnia (7% CO2) before and 1 hr after IP injections of saline, or 0.3 mg/kg or 1.0 mg/kg of the TRH analog Taltirelin (TAL). VE in RA did not differ among strains during all control studies, but VE increased after low and high dose TAL in both strains (p蠄0.05), with a greater increase in BN rats (p<0.05). The slope of the relationship between VE and PaCO2 breathing RA and 7% CO2 (ΔVE/ΔPaCO2) was lower in BN (0.4 ± 0.3) vs. SD (10.2 ± 1.5) rats after saline (p<0.05), and increased in BN (6.3 ± 1.0; p<0.05) but not SD (8.2 ± 1.2; p=0.15) rats after high dose TAL, where ΔVE/ΔPaCO2 no longer differed among the strains (p=0.207). TR also significantly increased in BN rats after high dose TAL (+1.1 ± 0.1°C; p<0.001), but was unaffected in SD rats (+0.1 ± 0.2°C; p=0.523). These data suggest an increased sensitivity to TRH receptor stimulation in BN rats, consistent with the concept that reduced endogenous TRH expression contributes to their blunted HCVR.Grant Funding Source: Supported by NIH HL097033
Generation of multiciliated cells in functional airway epithelium from human iPSC (1094.5)
Authors: Firth Amy , Menon Tushar , Dargitz Carl , Qualls Susan , Wright Rebecca , Khanna Ajai , Verma Inder 
Published: June 2021

Despite a significant improvement in the availability of therapeutic options to treat lung diseases, pulmonary disease still remains a major cause of morbidity and mortality around the world. Currently there are limited opportunities to study human lung disease either in vivo and in vitro. Using induced pluripotent stem cells (iPSC) we have generated a reproducible differentiation protocol to make mature post‐mitotic multiciliated cells in a functional airway epithelium.iPSC were generated from human skin biopsies and differentiated via FOXA2+SOX17+ definitive endoderm (>90% efficiency) to FOXA2+NKx2.1+ anterior foregut endoderm, FOXA2+NKx2.1+SOX2+ (~50% efficiency) pulmonary endoderm and then matured in an air liquid interface. Robust multiciliogenesis occurred when Notch signaling was inhibited and was confirmed by; i) the assembly of multiple pericentrin stained centrioles at the apical surface, ii) expression of transcription factor FOXJ1 and iii) presence of multiple acetylated tubulin labeled cilia projections in individual cells. The presence of NKx2.1+CC10+ Clara cells, MUC5A/C+ goblet cells and FOXA2+p63+ basal cells was also confirmed showing we are generating a complete polarized epithelial cell layer comprised of all relevant cell types. Functional cAMP activated and CFTRinh‐172 sensitive CFTR currents were recorded in isolated epithelial cells by whole cell patch clamp technique. Furthermore, we have corrected the deltaF508 mutation in the CFTR gene (>80% of all cases of CF) using a combination of CRISPR‐Cas9 endonuclease‐mediated genome editing and piggyBac transposase technologies, in the CF patient‐derived iPSC.The generation of mature multiciliated cells in a human iPSC differentiated respiratory epithelium and the ability to correct disease causing mutations provides a significant advancement toward modeling a number of human respiratory diseases in vitro.Grant Funding Source: Supported in part by CIRM and the Berger Foundation
The role of Na/K‐ATPase in cardiac stem cells (1094.6)
Authors: Shi Huilin , Drummond Christopher , Tian Jiang 
Published: June 2021

It has been shown that Na/K‐ATPase is involved in cardiac remodeling in 5/6th partial nephrectomized (PNx) mice and rats. Our recent work observed that genetic reduction of the Na/K‐ATPase α1 subunit results in higher cardiac c‐kit positive cell numbers. To delineate the mechanisms, we isolated sphere forming cardiac stem cells from Na/K‐ATPase α1 heterozygous mice and their wild type littermates to compare their proliferation capacity. It was found that heart tissue from α1 heterozygous mice produces more phase‐bright cells than that from WT mice during the same time period, indicating a faster growth rate with reduction of Na/K‐ATPase. The formation of first generation cardiospheres (CSps) on poly‐D‐lysine coated surfaces is also faster with cells isolated from α1 heterozygous mice. To test the proliferation rate in in vitro culture, the purified primary CSps derived cells (PCDCs) were obtained by growing first generation CSps on fibronectin coated surfaces. These cells show positive staining of progenitor cell markers c‐kit, Oct4 and Nkx2.5, but negative staining for the cardiac marker α‐actinin. The results showed that PCDCs from both wild type and heterozygous mice can form cardiospheres at a similar rate which are of a similar size. These data suggest that reduction of Na/K‐ATPase may regulate cell growth in vivo or in the early stage of stem cell purification but has no significant effect on its capacity of sphere forming once selected and cultured in vitro.Grant Funding Source: Suported by NIH grant R01 HL‐105649
Inhibitory effects of J10 on lung cancer cell motility: an in vitro and in vivo study (1095.14)
Authors: Wu Chieh‐Hsi , Chen Shuo‐Chueh , Chien Yi‐Chung , Pan Chun‐Hsu , Chen Chih‐Yi , Sheu Jyh‐Horng 
Published: June 2021

There are many major causes of cancer death, including metastasis of cancer. Dihydroaustrasulfone alcohol, which isisolated from marine coral, has shown antioxidant activity, but has not been reported to have an anti‐cancer effect. We first discovered that dihydroaustrasulfone alcohol provided a concentration‐dependent inhibitory effect on the migration and motility of human non‐small cell lung carcinoma (NSCLC) A549 cells by trans‐well and wound healing assays. The results of a zymography assay and Western blot showed that dihydroaustrasulfone alcohol suppressed the activities and protein expression of matrix metalloproteinase (MMP)‐2 and MMP‐9. Further investigation revealed that dihydroaustrasulfone alcohol suppressed the phosphorylation of ERK1/2, p38, and JNK1/2. Dihydroaustrasulfone alcohol also suppressed the expression of PI3K and the phosphorylation of Akt. Furthermore, dihydroaustrasulfone alcohol markedly inhibited tumor growth in Lewis lung cancer (LLC)‐bearing mice. We concluded that Dihydroaustrasulfone alcohol is a new pure compound with anti‐migration and anti‐tumor growth activity in lung cancer and might be applied to clinical treatment in the future.Grant Funding Source: Supported by National Science Council, Taiwan with the Grant No: NSC 101‐2320‐B‐039 ‐058 a