bluuebunny/crossref_metadata_2025_split huggingface dataset:
Atherosclerotic intralesional haemorrhage moulds monocyte differentiation into a macrophage antioxidant phenotype bearing the hemoglobin scavenger receptor CD163
Authors: Boyle Joseph J , Philippidis Pandelis , Horncastle Donna , Cantilena Sandra , Leung Viola , Taylor Kenneth M , Landis Clive , Haskard Dorian O 
Published: June 2021

Atherosclerotic intralesional haemorrhage promotes plaque progression. We investigated macrophage differentiation in atherosclerotic plaques with intralesional hemorrhage and in vitro with hemoglobin:haptoglobin (Hb:Hp) complexes. Macrophages within culprit lesions partitioned into reciprocal populations, with 26.1±2.3% expressing high levels of CD163 but depressed HLA‐DR (CD163high‐HLA‐DRlow phenotype), and the remainder 72.5±2.4% expressing the inverse (classical inflammatory) phenotype (CD163low‐HLA‐DRhigh). CD163high macrophages were associated with areas of hemorrhage (P<0.008), expressed increased (antioxidant) heme oxygenase‐1 (P<0.00001), and were protected from oxidant stress (P<0.003), despite greater iron content (P<0.001). This phenotype was reproduced in vitro by differentiating human monocytes with Hb:Hp, which required phagocytosis of Hb:Hp via CD163 and was inhibited by anti‐IL‐10. The divergent commitment of macrophages to CD163highor CD163low phenotypes was predictively modelled using a systems biology approach incorporating an IL‐10 autocrine positive feedback loop. Our results provide evidence for a novel CD163highmacrophage subset within culprit lesions. This appears to be moulded by the local microenvironment to become ‘fit for purpose’ for clearing Hb. Its therapeutic induction may reduce plaque progression.Funded by the British Heart Foundation
Coronin 1C negatively regulates cell migration in intestinal epithelial cells
Authors: Samarin Stanislav Nikolayevich , Nusrat Asma 
Published: June 2021

The coronin family of actin‐binding proteins consisting of seven proteins in mammals is known to play a critical role in various actin‐driven processes. Intestinal epithelial cells (IECs) abundantly express a member of coronin protein family, Coronin 1C. Since actin cytoskeletal restructuring is required for cell migration observed during pathologic processes, such as wound closure, we investigated the role of Coronin 1C in the regulation of IEC cell migration. Migration of a model intestinal epithelial cell line SK‐CO15 on different extracellular matrixes and cell invasion into a 3 dimensional (3D) matrix was determined. siRNA‐mediated knock‐down of Coronin 1C enhanced the formation of membrane protrusions in cells grown in 3D culture and increased their invasiveness into 3D matrix. Furthermore, the depletion of Coronin 1C significantly increased the rate of actin‐based motility in wounded SK‐CO15 cells by increasing the persistence of membrane protrusion at the leading edge of migrating cells. These findings support a negative regulatory role for Coronin 1C in IEC migration.This research is supported by the Crohn's and Colitis Foundation of America (for SNS) and NIH (DK55679 for AN).
Respiratory lethality in newborn NPAS3‐deficient mice
Authors: Zhou Shutang , Degan Simone , Sunday Mary E 
Published: June 2021

The neuronal PAS domain protein 3 (NPAS3) is a close homolog of Drosophila Trachealess. By quantitative RT‐PCR, NPAS3 gene expression is highest at E10.5‐E13.5 in developing murine lungs. NPAS3 immunostaining shows positive nuclei in the developing mesenchyme and epithelium. NPAS3 loss‐of‐function embryos (KOs) have deficient lung branching. More than 95% of NPAS3 KO newborns die within 48 hours after birth in respiratory distress with poorly developed alveoli, abnormal distribution of smooth muscle actin along the airways and an increase in intrapulmonary nerve fibers. NPAS3 Heterozygotes (NPAS3‐Hets) have reduced alveolarization at birth as well, which persists in adults. The 4‐month old NPAS3‐Hets show airway hyperreactivity (AHR). Similarly, reduced alveolarization and AHR occur in premature infants with chronic lung disease, called bronchopulmonary dysplasia (BPD). Microarrays and QRT‐PCR demonstrate significantly decreased expression of sonic hedgehog (SHH), Gli2, FGF9, and BMP4 as well as elevated expression of Sprouty2 in NPAS3 KOs. Deficits in any or all of these signaling pathways could contribute to the observed pulmonary lethal phenotype in NPAS3 KOs. Our data suggest that NPAS3 play an important role in lung development and in responses to lung injury.Supported by NIH Grant HL44984.
Inhibition of the soluble epoxide hydrolase (sEH) alters vascular remodeling
Authors: Revermann Marc , Barbosa‐Sicard Eduardo , Zwissler Bernhard , Fleming Ingrid , Hammock Bruce , Brandes Ralf Peter 
Published: June 2021

Epoxyeicosatrienoic acids (EETs) have potent anti‐inflammatory effects and promote endothelial function. The soluble epoxide hydrolase (sEH) metabolizes EETs to their less active diols (DHETs) and thus inhibition of sEH increases EET concentrations. Since EETs reduce smooth muscle cell (SMC) migration, proliferation and monocyte adhesion to the vascular wall in vitro, we hypothesized that inhibition of sEH using 12‐(3‐adamantan‐1‐yl‐ureido) dodecanoic acid (AUDA) prevents vascular remodeling in hyperlipidemic ApoE−/− mice. Remodeling was studied using models with different inflammatory and SMC‐stimulating properties. AUDA increased the ratio of EETs to DHETs in all models.The effect of AUDA in the femoral artery cuff model was a significant reduction of inflammation‐accompanied neointima within 2 weeks (43% reduction in intima/media ratio ‐ IMR, p<0,05). Carotid artery ligation induced a mainly SMC‐formed neointima within 4 weeks and this effect was significantly attenuated by AUDA (64% reduction in IMR, p<0,05).Angiotensin II infusion causes severe aortic inflammation aneurysm formation. In this model AUDA reduced thoracic aorta remodeling (26% less diameter, p<0.05). Taken together, these data demonstrate that sEH‐inhibition attenuates vascular remodeling ‐ probably via alteration of SMC‐activity and augmentation of anti‐inflammatory, the vascular wall protecting EET‐effects.Funding: German Research Foundation & J.W. Goethe‐University
A Water‐Soluble Formulation of HET0016 for the In‐Vivo Inhibition of 20‐HETE
Authors: Klamerus Megan Marie , Poloyac Samuel Mark , Rohan Lisa Cencia 
Published: June 2021

Purpose: N‐Hydroxy‐N’‐(4‐butyl‐2‐methylphenyl)‐formamidine (HET0016) is a potent inhibitor of the cytochrome P450 enzymes responsible for the formation of the small vessel vasoconstrictor 20‐Hydroxyeicosatetraenoic acid (20‐HETE). However, HET0016 has a low aqueous solubility, limiting in vivo administration. The aim of this study was to use 2‐hydroxypropyl β‐cyclodextrin (2‐HPβCD) to create a water‐soluble formulation of HET0016.Methods: The water solubility of HET0016 in both ddH2O and 15% 2‐HPβCD was determined. Rats then received 15% 2‐HPβCD or a formulation of HET0016 and 2‐HPβCD. Brain cortex, kidney and liver were subsequently analyzed for 20‐HETE content via HPLC/MS‐MS. Additional studies were also performed in order to maximize the amount of HET0016 and minimize the amount of 2‐HPβCD in the formulation.Results: The solubility of HET0016 in ddH2O was found to be 34.2μg/mL compared to 452.7μg/mL in 15% 2‐HPβCD. 20‐HETE concentrations for the control group were 38.5, 189.2 and 835ng/g tissue in the brain cortex, kidney and liver, respectively. 20‐HETE concentrations in the same tissues for the experimental group were 9.8, 37.4 and 231.7ng/g tissue, respectively. Optimization of the HET0016:2‐HPβCD formulation showed that the greatest amount of HET0016 present in the smallest percentage of 2‐HPβCD was 1.3mg in 12.5%.Conclusions: 2‐HPβCD increased the water solubility of HET0016 thirteen‐fold and also provided a water‐soluble formulation of this compound. When administered to rats, 20‐HETE formation was inhibited by 74.7% in the brain cortex, by 80.2% in the kidney and by 72.3% in the liver.This work is supported by the NIH.
Topical Dual VEGF Receptor/Src Kinase Inhibitors as Potential Therapies for Ischemic Retinal Vascular Disorders
Authors: Mahesh Sankaranarayana Pillai , Virata Cyrus A , Chen Zoe , Cao Jon , Mak Chi Ching , McPherson Andrew , Noronha Glenn , Soll Richard 
Published: June 2021

Purpose: Ischemic retinal vein occlusion (RVO) is a common retinal vascular disorder and carries a high risk of visual loss with few therapeutic options. We evaluated whether dual inhibition of VEGFR and down‐stream Src family tyrosine kinases using small molecules is a potential therapeutic intervention for ischemic retinal vascular disorders.Methods: We examined two distinct chemical classes of dual VEGFR/Src kinase inhibitors (represented by compounds TG100801 and TG100948) in a rat model of RVO for their ability to impact retinal edema. Ischemic retinal vein occlusion was induced in rat retina by thrombosis of a single retinal vein using argon laser photocoagulation following injection of a photo‐sensitizer dye. Edema was quantified either using scanning optical fluorophotometry, measuring fluorescein leakage in to the retina, or by optical coherence tomography (OCT), measuring retinal thickness.Results: Following topical administration, both the compounds achieved substantial levels in the retina and intraocular tissues with little to no systemic exposure. At 48 hrs post‐vein occlusion, both TG100801 and TG100948 successfully reduced edema compared to vehicle (by 47% with 0.3% TG100801, P= 0.002, and by 38% with 0.03% TG100948, P= 0.039). OCT scans also showed reduced retinal thickening as well as resolution of pockets of bullous edema.Conclusions: As VEGF plays a key role in the edema and neovascularization which develop in the ischemic retina, we propose that multi‐targeted inhibitors of the VEGF signaling cascade represent promising therapies for these vascular disorders. Moreover, as current forms of therapy require invasive delivery techniques (intravitreal injections), compounds such as TG100801 and TG100948, which can be topically delivered, offer additional therapeutic advantages.
Scavenger receptor BI prevents septic death
Authors: Li Xiang‐An , Guo Ling , Song Zhiqing , Smart Eric J 
Published: June 2021

Scavenger receptor BI (SR‐BI) is a HDL receptor which regulates plasma HDL cholesterol levels by selectively uptake cholesterol ester from HDL. We recently reported a novel function of SR‐BI namely protection against endotoxin induced animal death. Using a lipopolysaccharides (LPS) induced endotoxic animal model, we found that LPS challenge induced 90% fatality of SR‐BI null mice while all the wild type littermates survived, indicating that SR‐BI plays a pivotal role in protection against LPS toxicity in vivo. In the current studies, we determined whether SR‐BI plays a protective role in sepsis using an established septic animal model (cecum ligation and puncture, CLP). We demonstrate that CLP treatment induced 100% fatality of SR‐BI null mice within three days (n=20) whereas only 21% fatality of wild type littermates (n=27) and 36% fatality of heterozygous littermates (n=30). Our findings clearly demonstrate that SR‐BI is a critical protective modulator in sepsis in mice, which may reveal a novel target for the intervention of sepsis.(Supported by grants from American Heart Association 0530241N and from the NIH 2P20RR015592)
Role of Egfl7 During Vascular Development
Authors: Stuhlmann Heidi , Durrans Anna , Nichol Donna , Campagnolo Luisa 
Published: June 2021

Epidermal growth factor domain‐like 7, Egfl7, encodes a 29kDa protein with an early and endothelial‐restricted expression profile. In vivo, Egfl7 mRNA is detected at mouse embryonic day 7.5. During development, expression is restricted to sites of blood island formation, the vascular endothelium, and endocardium. In adults, Egfl7 is down‐regulated except in the lung, pregnant uterus, and regenerating endothelium. We have used loss‐ and gain‐of‐function to determine the role of Egfl7 during vasculogenesis and angiogenesis. We are using a lentiviral siRNA‐mediated approach to establish Egfl7 knock‐down in ESC clones and transgenic mice. Embryoid bodies with Egfl7 knock‐down appear to have less organized vascular cords compared with controls. Tie2‐Egfl7 transgenic mice, with forced expression of the gene in endothelial cells, have been generated for gain‐of‐function analysis, and these display increased vessel lumen diameter, hemorrhaging, and other vascular abnormalities. We will report on analyses of these ESC and transgenic mice studies, and describe how they relate to the role of Egfl7 during embryogenesis. In addition, we will report on a second site of Egfl7 expression in embryonic germ cells (EGC) and embryonic stem cells (ESC), primordial stem cells, and during adult spermatogenesis.Research Support: NIH HL082098, Tri‐Institutional Stem Cell Initiative (H.S.)
Chondrogenic differentiation of MSC expanded under FCS‐free inflammatory condition
Authors: Felka Tino , Schäfer Richard , Schewe Bernhard , Kluba Torsten , Aicher Wilhelm K 
Published: June 2021

Objectives:Mesenchymal stromal cells (MSC) are a promising tool for tissue engineering of the intervertebral disc. The injured tissue is usually characterized by inflammatory processes. In this study we investigated the effect of IL‐1beta on the chondrogenic differentiation of MSC.Methods:Bone‐marrow‐derived MSC were cultured in FCS‐free, GMP‐compatible medium and characterized by flow cytometry. Chondrogenic differentiation of MSC was induced with and without IL‐1beta supplementation. After 28 days, micromasses were analyzed by immunostaining. In addition, the gene expression of several matrix proteins was investigated by quantitative PCR.Results:Micromasses differentiated under IL‐1beta supplementation are characterized by smaller dimensions, disruptions and weaker accumulation and expression of extracellular matrix proteins compared to micromasses differentiated without IL‐1beta supplementation.Conclusion:Inflammatory processes in the injured tissue could inhibit the chondrogenic differentiation of transplanted MSC. For cell therapy to be effective it will be necessary to develop strategies to ensure the steady expression of extracellular matrix proteins under inflammatory conditions.
A Composite Scaffold for the Engineering of Hollow Organs and Tissues
Authors: Eberli Daniel , Yoo James J. , Atala Anthony 
Published: June 2021

IntroductionSeveral types of synthetic and naturally derived biomaterials have been used for augmenting hollow organs and tissues. However, each has desirable traits which were exclusive of the other. We fabricated a composite scaffold and tested its potential for the engineering of hollow organs in a bladder tissue model.Materials and MethodsThe composite scaffolds were configured to accommodate a large number of cells on one side and were designed to serve as a barrier on the other side. The scaffolds were fabricated by bonding a collagen matrix to PGA polymers with threaded collagen fiber stitches. Bladder cells were seeded on the composite scaffolds, and implanted in mice for up to 4 weeks and analyzed.Results and DiscussionThe composite scaffolds, consisting of an acellular matrix and PGA polymers, are tightly bonded, and maintained their structural properties. The bladder cells readily attached and proliferated on the scaffolds and formed bladder tissue structures in vivo. The biomechanical studies demonstrated that the tissues were readily elastic while maintaining their pre‐configured structures.ConclusionsWe demonstrate that a composite scaffold can be fabricated with two completely different polymer systems for the engineering of hollow organs.
RGS2 is a negative regulator of Gq/11‐mediated signaling and cell proliferation in adult rat ventricular fibroblasts
Authors: Zhang Peng , King Michelle , Ayrapetov Marina , Mende Ulrike 
Published: June 2021

Cardiac fibroblasts (CF) contribute to cardiac remodeling by assuming an “activated” myofibroblast (MyoFb) phenotype. Many G protein‐coupled receptors are involved in CF activation. G proteins are controlled by Regulators of G protein Signaling (RGS). RGS expression and function in CF and MyoFb are unknown. We set out to identify which RGS are expressed in isolated adult rat ventricular CF (passage P0‐P2) and to begin to delineate their function. Morphologic changes and expression of MyoFb markers (e.g., α‐smooth muscle actin) indicated gradual conversion from CF (P0) to MyoFb (P2). We detected mRNA of 14 RGS by RT‐PCR that were differentially regulated during CF to MyoFb conversion. Only RGS2 mRNA was markedly up‐regulated in response to short‐term stimulation of Gq/11‐coupled Angiotensin II (Ang II) and Gs‐coupled β‐adrenergic receptors. Adenoviral RGS2 expression blunted Ang II‐induced phospholipase C β activation by 65±2% (P0), 66±2% (P1) and 78±3% (P2) (n=3, P<0.05) and CF proliferation (BrdU incorporation) by 37±5% (n=10, P<0.05). RGS2 had no effect on Gs‐coupled cAMP accumulation. Thus, while RGS2 is selectively up‐regulated in response to Gq/11 and Gs activation, it negatively regulates only Gq/11‐mediated signaling and proliferation in both CF and MyoFb. These data point to RGS2 as an important regulator of CF and MyoFb signaling and function.Supported by the NIH and American Heart Association.
Isoform Specific Regulatory Site for PKA: AGC‐Specific aH‐aI Loop
Authors: yang jie , Kenndy Eileen , Wu Jian , Deal Michael , Brown Simon , Pennypacker Juniper , Ghosh Gourisankar 
Published: June 2021

In an initial study of ATP binding site in the catalytic subunit of PKA, a distantly coupled mutation pair of F327A/K285P was identified through a yeast genetic screen. It was found that while the wild‐type mammalian C‐subunit can sustain growth in a yeast strain devoid of PKA‐expressing tpk genes, the F327A mutant, a C‐terminal residue that constitutes part of the ATP binding site, failed to do so. Screening for suppressors by means of restored cell survival in the tpk‐null strain identified several suppressor mutations, one of which was K285 to Pro. K285 lies in an AGC‐specific insert between the aH‐aI helices of the large lobe of the kinase domain. In this study, we confirmed an important role of F327 in stabilizing ATP binding. We also showed that the second site mutation of K285P did not restore PKA activity; rather, through allosteric connection with F327A, the double mutant exhibited decreased inhibition by the inhibitory regulatory subunit of PKA. Thus a low yet constant PKA activity was maintained which appeared to be sufficient for cell viability.Further characterization of the mutants unexpectedly showed that K285 constitutes an interacting site that is critical for the RII‐, but not the RI‐ subunit of PKA; this isoform‐specific interaction was not previously appreciated. This study highlighted a unique feature of the K285 site in the AGC specific insert, which may have considerable relevance to other AGC kinases.
A Central Role for CD36 in Mediating Oxidized LDL and Fatty Acid Induced Insulin Resistance
Authors: Kennedy David J , Kuchibhotla Sai , Vanegas Difernando , Nimako George , Febbraio Maria , Silverstein Roy L 
Published: June 2021

Elevated levels of oxidized LDL (oxLDL) and fatty acids (FA) are highly associated with insulin resistance. Clinical and experimental evidence also demonstrate increased expression of scavenger receptor CD36 in insulin resistant states. As oxLDL and FA are CD36 ligands, we tested the hypothesis that they influence insulin resistance in a CD36 dependant manner.3T3‐L1 adipocytes treated with oxLDL or with FA showed time and dose dependent increases in CD36 expression, activated Jun kinase (JNK), induced inhibitory IRS‐1 serine phosphorylation, increased adipocyte lypolysis and decreased glucose uptake. Pre‐incubation with either a CD36 monoclonal antibody or JNK kinase inhibitor attenuated these changes.Next, ApoE null mice with or without a CD36 null mutation (CD36+ or CD36−) were fed a high fat diet for 12 weeks and examined for physiologic and biochemical markers of insulin resistance. These mice have increased oxLDL and FA levels and their isolated lipoproteins are CD36 ligands, inducing CD36‐dependent foam cell formation and atherosclerosis. Compared with CD36+ mice, CD36− mice had improved glucose clearance, decreased JNK activation and IRS‐1 serine phosphorylation in adipose tissue, increased AKT activation in both liver and muscle, as well as increased levels of adiponectin in adipose, muscle and liver.These data suggest a central role for CD36 in mediating oxLDL and FA induced insulin resistance.
Regulation of Cytokinesis by GTPase Flux
Authors: Miller Ann Louise , Bement William 
Published: June 2021

How do microtubules of the mitotic spindle specify the actomyosin‐based contractile ring during cytokinesis? Prior to contractile ring assembly, spindle microtubules precisely localize proteins that regulate the small GTPase Rho to the cell equator. These Rho regulators then direct formation of a zone of high Rho activity, which promotes actomyosin contractility. We are examining the roles of the Rho regulator MgcRacGAP during cytokinesis in Xenopus embryos. We find that MgcRacGAP is required for cytokinesis, as knock‐down of MgcRacGAP leads to multinucleate cells. This multinucleate phenotype can be rescued by re‐expressing wild type MgcRacGAP but cannot be rescued fully by either of two GAP‐DEAD MgcRacGAP mutants. Expression of the GAP‐DEAD mutants leads to Rho activity zones that are broad and spatially unstable compared to controls, suggesting that GAP activity is necessary to maintain a focused Rho activity zone. Additionally, disruption of MgcRacGAP's GAP activity promotes increased accumulation of downstream targets of Rho at the furrow. We propose that during cytokinesis the Rho activator Ect2 locally activates Rho, while MgcRacGAP locally inactivates Rho, resulting in the constant flux of Rho through the GTPase cycle. By using the GTPase flux mechanism, cells can maintain a tightly focused zone of Rho activity, which is necessary to drive contractile ring formation.Funding: American Cancer Society and Helen Hay Whitney Postdoctoral Fellowships to ALM. NIH grant to WMB.
The effects of pro‐apoptotic DNA vaccinations on Dendritic Cells
Authors: Perez Jessica Marie , Li Alice , Escher Alan 
Published: June 2021

Pro‐apoptotic DNA vaccines specifically pND2‐SGAD55‐BAX, have been found to suppress diabetes and insulitis in non‐obese diabetic (NOD) mice, ameliorate new‐onset diabetes, and induce foxp3 regulatory T cells in pre‐diabetic mice with early diabetes diagnosis.(1,2)The effects of the apoptotic DNA vaccines are now being studied. This study focuses on the effects of apoptotic DNA vaccines on Dendritic cells (DCs). This study will focus on effects of the vaccines on DCs gene expression level of mRNA. Two methods of transfection were used to introduce three different plasmid constructs, BAX, GAD65‐BAX, and SGAD55‐BAX. These methods were lipofection and electroporation. The results of the Real Time Polymerase Chain Reaction (RT‐PCR) were used to measure the mRNA expression level of seven different genes including: CD40, CD80, CD86, TGFβ1, FCγIIB, IL‐1RA, and GAPDH, as house‐keeping gene control. Results showed that the delivery of SGAD55‐BAX induced significant changes in gene expression in DCs when compared to GAD65‐BAX. DCs transfected via lipofection with the SGAD55‐BAX caused decreased expression of CD80, CD86, TGFβ1, FCγIIB, and IL‐1RA compared to the delivery of BAX alone. However delivery of the same plasmid via electroporation showed increased expression of the same genes compared to BAX alone. These results may be due to different efficiencies of transfection of plasmid DNA or due to the capacity of DCs to respond rapidly to different stimuli.National Institute of Health‐National Center On Minority Health and Health Disparities through grant 5P20MD001632 (LLU Center for Health Disparities Research)