bluuebunny/crossref_metadata_2025_split huggingface dataset:
Resolution of Right Ventricular Fibrosis by a Cancer Chemotherapeutic Agent: A Novel Therapy to Treat Pulmonary Arterial Hypertension
Authors: Rybka Vladyslava , Shults Nataliia V. , Ibrahim Yasmine F. , Suzuki Yuichiro 
Published: November 2023

Pulmonary arterial hypertension (PAH) remains a fatal disease despite the availabilities of FDA‐approved vasodilators. Since right heart failure is the major cause of death among patients with PAH, new agents that can repair the damaged right ventricle (RV) have therapeutic potential. We herein report that a clinically used anti‐cancer drug, docetaxel, is capable of resolving RV fibrosis and repairing the damaged RV. Treatment of rats with SU5416 and chronic hypoxia caused PAH and severe RV damage including the occurrence of fibrosis as indicated by the accumulation of collagen. The treatment of these PAH rats with docetaxel after RV damages had developed resulted in a significant reduction of RV fibrosis with increased contractile functions. Treatment of cultured alpha‐smooth muscle actin‐expressing adult human ventricular myofibroblasts with docetaxel caused the cell death. Further, these myofibroblasts were found to have downregulated collagen 1. These results demonstrate that the docetaxel treatment results in the resolution of fibrosis and the restoration of functional cardiomyocytes in the damaged RVs in PAH animals. Mechanisms of docetaxel actions may involve the killing of myofibroblasts as well as inhibiting the collagen synthesis or promoting the collagen degradation. Targeting RV fibrosis should be considered as a therapeutic strategy for treating pulmonary arterial hypertension patients.Support or Funding InformationNIH
Phospholipase D Regulates GSK3β Mediated Epithelial to Mesenchymal Transition and Akt Mediated Cell Death Leading to Pulmonary Fibrosis
Authors: Suryadevara Vidyani , Royston Thomas J. , Natarajan Viswanathan 
Published: November 2023

Idiopathic pulmonary fibrosis (IPF) is a pernicious lung disease characterized by scar formation and respiratory failure. This is due to dysregulated repair of the epithelial injury in the lung, which involves epithelial apoptosis and also epithelial to mesenchymal transition (EMT), thereby leading to the deposition of the extracellular matrix in the lung. Our previous studies have established a role for phospholipase D (PLD) as genetic deletion of PLD2 in mice conferred protection against bleomycin induced PF. PLD2−/− mice exhibited less inflammation on day 7 and less pulmonary fibrosis on day 21 post‐bleomycin challenge compared to wild type mice. This study aims at understanding how PLD regulates GSK3β mediated EMT and Akt mediated apoptosis. GSK3β and Akt are known to be involved in crucial signaling pathways that regulate cell death and survival. To elucidate the role of PLD signaling in epithelial injury, Beas2B cells were pre‐incubated with either PLD1 (VU0155069‐ 250nM), PLD2 (VU0364739‐500nM) or PLD1 +PLD2 inhibitors prior to bleomycin challenge. Bleomycin challenge enhanced phosphorylation of GSK3β at Ser9 leading to its inactivation, which was reversed by inhibition of PLD1 and PLD2. Also, activated Akt as shown by enhanced phosphorylation at Ser473 was reversed upon PLD1 and PLD2 inhibition. Inhibition of PLD1 and PLD2 was necessary to attenuate bleomycin mediated apoptosis of Beas2B cells as evidenced by cleavage of Caspase‐3 and PARP. The results presented here suggest that PLD regulates bleomycin‐induced pulmonary fibrosis by modulating apoptosis and EMT via Akt/GSK3β signaling in the epithelial cells, and targeting PLD may be beneficial.Support or Funding InformationThis work is supported by National Institutes of Health grant P01 HL 98050 to VN.
Detection of Phenotypic Differences in Alveolar Macrophages Using Ionized Calcium‐Binding Adapter Molecule 1 Marker
Authors: Meyerholz David K , Hogan Christin M. , Glanz Ryan M , Goeken J Adam , Leidinger Mariah R , Ofori‐Amanfo Georgina K , McQuillen Logan , Donovan Kathleen M , Harwani Sailesh C , Flaherty Heather A. 
Published: November 2023

Alveolar macrophages are resident immune cells in the distal lung and offer a first line of defense to inhaled pathogens and environmental particulates. In healthy states, these resident immune cells reside in the alveolar spaces of the lung as quiescent monitors, but upon stimulation, they play significant roles in host defense, removal of inhaled particulates and tissue remodeling. Upon activation, alveolar macrophages can exhibit morphological changes including increases in cell size, ruffled borders and altered expression of markers. In this study of 10 week old male rats, we had immunostained lung sections for the marker, ionized calcium‐binding adapter molecule 1 (IBA1). We evaluated whether quantitative or qualitative differences could be detected in alveolar macrophages between the normotensive WKY (Wistar Kyoto) strain and the hypertensive SHR (Spontaneous Hypertensive Strain) strains. For all analysis, statistical significance was set at P<0.05. Immunocytochemistry of bronchoalveolar fluid showed similar immunostaining of macrophages in both groups with no overt group‐distinguishing features. However, examination of immunostained paraffin embedded tissue sections suggest that SHR lungs have subpopulations of macrophages with increased cell size. We evaluated macrophage size in these subpopulations using a semiquantitative scoring system and then by quantitative evaluation of macrophage diameter with both showing significant increases in SHR macrophages (P<0.01 for both). To test the extent of IBA1 immunostaining in lung high resolution images were quantified. SHR lungs had more IBA1 immunostaining than WKY (P<0.05). Interestingly, IBA1 immunostaining in small, quiescent macrophages was often homogenous throughout the cytoplasm whereas macrophages that were larger in size often had a central zone of cytoplasmic pallor with stronger immunostaining near the periphery/membrane of the cell. These findings suggest that quantitative and qualitative features of IBA1 immunostaining may be useful tools to detect differences in the activation state of alveolar macrophages.Support or Funding InformationThis work was supported in part by the NIH
Sex Dependent Role of Activating Transcription Factor 3 (ATF3) Expression In Modulating The Asthmatic Phenotype In An Acute Mouse Model Of Airway Neutrophilia
Authors: Spinelli Amy , Ortiz Nathalie Fuentes , Nicoleau Marvin , Sinha Utkarshna , Caruso Carla R , DiAngelo Susan , Chroneos Zissis , Silveyra Patricia 
Published: November 2023

BackgroundAsthma is a chronic lung disease that affects more than 300 million people worldwide, causing more than 250,000 annual deaths. A sub‐population of severe asthma patients have been identified that are distinguished by their high incidence of airway neutrophilia. Patients presenting with this inflammatory profile tend to be less responsive to corticoid steroid treatment and have a higher mortality rate. Therefore, identifying molecules that promote this excessive neutrophil recruitment is essential for better treatment options for these patients. In this study, we aimed to generate and characterize an acute mouse model of airway neutrophilia that could be used to study this particular asthma phenotype.MethodsBoth male and female C57BL/6J mice were challenged with either filtered air (FA) or ozone (1 or 2ppm for 3h). The asthmatic phenotype was assessed by testing airway hyperresponsiveness (AHR) to methacholine challenge using the Buxco system (Penh) and airway remodeling was characterized using the multi‐photon imaging technique‐second harmonic generation to determine collagen content in challenged airways. The extent of airway inflammation was quantified by cell differential assessment of bronchoalveolar lavage fluid (BALF) and lung tissue was harvested for mRNA expression measurements.ResultsWe found that ozone exposure leads to airway neutrophilia and AHR. We also saw that expression levels of the stress response activating transcription factor 3 (ATF3) were enhanced in ozone challenged animals with females showing a higher level of up‐regulation compared to males. Interestingly, global knockout of ATF3 was not protective against airway neutrophilia as ATF3−/− male and female mice were observed to have higher levels of neutrophil BALF counts, and female ATF3−/− mice had higher Penh values as compared to wild‐type ozone challenged mice.ConclusionThis study presents an acute model of refractory asthma whereby ozone challenge initiates AHR and airway neutrophilia. This study also demonstrates the complex role ATF3 plays in regulating airway neutrophilia whereby up‐regulation of this transcription factor is observed to promote the inflammatory phenotype but loss of this protein enhances this inflammatory response in a sex dependent manner.Support or Funding InformationThe authors also thank Todd Umstead for assistance, training and experiments. Dr. Joanna Floros for use of her ozone equipment. NIH Grant KO1HL133520
Inflammation Response of Phage‐Based Films on Titanium Surface in Vitro
Authors: Sun Yuhua , Feng Bo 
Published: November 2023

Phage‐based materials exhibit great potential in the applications of tissue regeneration. However, challenges still exist in the development of this field, such as a good understanding of the immune response to phage‐based biomaterials. The objective of this study was to examine the inflammation response of phage film and phage films chemically grafted peptides in vitro. Three different peptides with different affinity to transforming growth factor β1 (TGF‐β1) were used. Macrophage (RAW 246.7) adhesion, proliferation, apoptosis, polarization and expression levels of pro‐inflammatory cytokines, IFN‐γ, TNF‐α, IL‐6, IL‐1β, and anti‐inflammatory cytokines, IL‐10, IL‐4, TGF‐β1, were evaluated. The results showed that the phage films on Ti surfaces would induce the early apoptosis of the adherent macrophage and reduce the proliferation rate. In late culture period, a small amount of cell aggregation was present, but no foreign body giant cells were found. These films promoted sequential M1 and M2 polarization, as measured by expression of surface markers (CCR7 and CD206) and secretion of cytokines. The third day was the turning point of the polarization with increasing secretion of anti‐inflammatory cytokines, IL‐10 and TGF‐β1, although the ratio of M1 polarization was the greatest on the fifth day. No secretion of IFN‐γ and IL‐4 were detected. Macrophage polarization was primarily co‐regulated by phage films and the secreted factors (IL‐10, TGF‐β1 and TNF‐α). Compared to the control group, phage films downregulated the inflammatory response. In addition, the phage film grafted peptide with greatest affinity to TGF‐β1 induced higher early apoptosis of adherent macrophage and IL‐10 expression. The association with the binding of active TGF‐β1 and adsorption of serum proteins will be further studied in detail. The results of this study provide highly encouraging evidence for the safety of potential application of phage‐based materials in vivo.Support or Funding InformationNatural Science Foundation of China (31570955)
Profiling of inflammatory biomarkers in synovial fluid from patients undergoing primary total joint arthroplasty
Authors: Finkler Elissa , Wanderling Christopher , Thorson Chase , Galicia Kevin , Statz Stephen , Fareed Jawed , Hopkinson William 
Published: November 2023

IntroductionSynovial fluid is an intra‐articular fluid whose main function is to lubricate joints and decrease the coefficient of friction. Synovial fluid contains various proteins, such as lubricin, which contributes to maintaining joint homeostasis and preserving articulating cartilage. The role of inflammatory biomarkers in degenerative joint disease (DJD) has been well established. Previous reports have suggested that increased levels of biomarkers such as IL‐1b, CRP, and MP‐TP play a role in the mediation of inflammatory responses. It is hypothesized that the levels of these biomarkers may provide additional information on the pathogenesis of DJD. This study is designed to profile synovial fluid samples for these biomarkers.Materials and MethodsThis study consisted of 25 patients undergoing primary total joint arthroplasty (TJA) of the hip or knee for DJD. Procedures were performed by one of four attending orthopaedic arthroplasty surgeons. Patients were consented prior to surgery in accordance with LUMC IRB. At the time of arthrotomy, a sample of synovial fluid was obtained using a syringe and placed into a tube containing calcium citrate for preservation and analysis. Biomarkers including IL‐1b, MP‐TF and CRP were measured using commercially available ELISA methods from R&D laboratories (Minneapolis, MN). Total protein levels were measured by nantrope technology.ResultsTotal protein levels ranged from 5–65 mg/ml (mean 18 mg/ml +/− standard deviation 13 mg/ml). IL‐1b levels ranged from 0.04–0.96 pg/ml (0.19pg/ml +/−0.18pg/ml). MP‐TF levels ranged from 0–55 pg/ml (14pg/ml+/−19pg/ml). CRP levels ranged from 0–7 ug/ml (1.5 ug/ml +/− 1.9ug/ml).DiscussionThe results from this study demonstrate that synovial fluid from patients with DJD contains detectable levels of inflammatory biomarkers. The presence of IL‐1b and CRP is highly suggestive of an ongoing inflammatory process which may hold significance in the pathophysiology of DJD whereas MP‐TF levels indicate similar degradation products and is a thrombotic biomarker in the form of microparticles complexing with tissue factor. The wide variation in the levels of these markers may suggest differing degrees of degenerative and inflammatory processes. These levels can be quantified to provide more insight into the role of these mediators in the pathogenesis of DJD.
Comparative analysis of inflammatory cytokines and growth factors in young and old aged normal individuals
Authors: Rodrigues Sandra Maria , Walborn Amanda , Hoppensteadt Debra , Fareed Jawed , Rondina Matthew 
Published: November 2023

IntroductionHuman aging is a complex phenomenon, characterized in part by chronic, low‐grade inflammation. This increased inflammation, known as “inflammaging”, is a risk factor for increased morbidity and mortality, as most of the diseases associated of aging involve an inflammatory pathogenesis. Accordingly, there is a need to understand the inflammation associated with aging in order to develop therapies targeting inflammaging that may have benefits in the elderly population.Specific AimTo compare the levels of cytokines in young (ages 18–55) and old aged (ages 55–85) normal individualsMaterials and MethodsAn Evidence investigator cytokine and growth factor biochip array (RANDOX laboratories Ltd.,) was used to perform simultaneous quantitative detection of multiple analytes namely IL‐1α, IL‐1β, IL‐2, IL‐4, IL‐6, IL‐8, IL‐10, VEGF, TNF‐α, IFN‐γ, EGF, and MCP‐1. The Randox biochip is a solid substrate with multiple specific cytokine antibodies attached at pre‐defined sites on the surface. Each cytokine is bound to its specific antibody and then appropriate enzyme‐labeled secondary antibodies are added. The cytokines are quantified by chemiluminescence using a charge‐coupled device camera and imaging system. The concentration of the analyte present in the sample was calculated from the calibration curveResultsThe levels of cytokines VEGF, TNFα, MCP1 and EGF (p>0.05) were observed to be significantly higher in the old aged normal individuals when compared to that in the young aged normals. Two tailed unpaired t test was used to determine whether there were any significant differences between the mean cytokine levels of the two groups and standard error was used for the error bars.ConclusionThe elevated levels of cytokines in the old aged patients could be a risk factor for morbidity and mortality. With age the damaged organelles, cells and macromolecules tend to accumulate and form host derived endogenous cell debris. There is also increased activation of coagulation system and immunosenescence on aging. All these may contribute as sources of inflammaging stimuli. Therefore, these cytokines can be used as biomarkers for the diagnosis and disease management of various age related pathogenesis.
Gelsolin, but not mucin, inhibits P. aeruginosa swimming motility in vitro
Authors: Husak Alexander B. , Busch Sydney A. , Robarge Jordyn , Bankers‐Fulbright Jennifer L. 
Published: November 2023

Pseudomonas aeruginosa is a ubiquitous, opportunistic pathogen responsible for substantial morbidity and mortality in patients with cystic fibrosis (CF). Following initial colonization of the CF lung, P. aeruginosa changes phenotype and becomes virtually impossible to inhibit, kill, or remove. Thus, understanding and preventing initial P. aeruginosa colonization is of great interest. Our lab has previously demonstrated that apical secretions from a human airway cell line, Calu‐3, inhibit P. aeruginosa swimming motility and biofilm formation in vitro, and that this inhibition is lost if proteins > 50 kDa are removed. However, secretions from Calu‐3 cells lacking the CFTR (CF‐like cells) do not inhibit P. aeruginosa. The goal of this project is to identify the inhibitory protein(s) present in non‐CF apical airway secretions. Using reducing SDS‐PAGE, we identified a band at approximately 90 – 100 kDa that was present in normal apical airway secretions but not CF‐like secretions, suggesting that a protein this size may be our target. We have begun testing readily available candidate proteins – mucin and gelsolin ‐‐ to see if they can replicate the inhibitory activity of non‐CF airway secretions. Mucin had no effect on P. aeruginosa swimming motility or biofilm formation, but gelsolin significantly inhibited swimming motility in a dose‐dependent manner. Interestingly, P. aeruginosa biofilm formation was not inhibited by gelsolin. Thus, gelsolin may be responsible for at least a part of the P. aeruginosa inhibition induced by apical airway secretions.Support or Funding InformationFunding provided by Augsburg College Undergraduate Research and Graduate Opportunities Office and the Augsburg College Biology Department
Better targeting, better efficiency of Adeno‐associated virus gene transfer in the Central Nervous System for expression of Retromer proteins
Authors: Porter Demisha D.L , Jackson Kasey L , Dayton Robert D , Klein Ronald L 
Published: November 2023

Researchers are using AAV gene transfer more and more in their neuroscience studies, including optogenetics, cre‐lox targeting, and CRISPR gene editing. Our lab has been using the AAV9 vector serotype to express genes on a wide‐scale basis throughout the central nervous system in order to study a disease that affects cells throughout the central nervous system, amyotrophic lateral sclerosis. This current project explored ways to achieve more efficient expression and in a more targeted manner than our previous work. The recently described AAV PHP.B serotype was administered to rats for the first time, and it did result in significantly more green fluorescent protein expression than did the AAV9 vector in a systematic comparison. In studies comparing the neuron‐specific synapsin promoter with our previous cytomegalovirus/ chicken β‐actin promoter, the latter promoter was noticeably more efficient yet the synapsin promoter did yield a transduction pattern with greater neuron‐selectivity, which was the goal. We also explored intracerebroventricular injections to achieve wide‐scale transduction and better avoid unwanted transduction of peripheral organs. We are combining the synapsin promoter in AAV PHP.B vectors to take advantage of these efficiency and targeting features. Our specific genes of interest that we wish to express are called the retromer proteins. Retromers make up a protein complex that is associated with the trafficking of cellular vesicles through the endosome and the trans‐Golgi network. Viral constructs were made for epitope tagged versions of the retromers VPS29 and VPS35. We hypothesize that the retromers can act to prevent the formation of beta‐amyloid plaques in Alzheimer's disease by altering the trafficking of amyloid precursor protein.Support or Funding InformationDr. Ronald Klein, Kasey Jackson, Robbie Dayton, Fidelity Biosciences Research Initiative, Karyopharm Therapeutics, Inc., ALS Association, Meira GTx, The LSU Health Sciences Center Foundation in Shreveport, The American Society for Pharmacology and Experimental Therapeutics, The Society for Toxicology
Surface area may be a more useful risk factor for cerebral aneurysm rupture than maximum diameter
Authors: Fukuda Shunichi , Shimogona Yuji 
Published: November 2023

IntroductionThe maximum diameter of unruptured cerebral aneurysms is the most significant risk factor of aneurysm rupture. The larger the aneurysm, the easier it is to rupture, and a size of 7 mm or more is clinically significant. This is thought to be at least partially due to hemodynamic involvement. We show here associations between hemodynamic factors and maximum diameter and surface area of cerebral aneurysms by regression analysis.MethodsWe are conducting a multi‐institutional prospective observational clinical study to examine hemodynamics of human aneurysm development, enlargement, and rupture using computational fluid dynamics (CFD); CFD ABO Study. Three‐dimensional CT angiographic images and flow velocities were acquired from 74 patients in the Study.ResultsWSS, NWSS, and WSSG, which are metrics for wall shear stress, at the maximum diameter of 7 mm or more and the surface area of 40 mm2 or more are significantly lower than these at the maximum diameter less than 7 mm and the surface area less than 40 mm2, respectively. Similarly, OSI, GON, and NtransWSS, which are metrics for disturbed flow, at the formers are significantly higher than those at the latters. In addition, the associations between surface area and NWSS, WSSG, GON, or NtransWSS are significant (regression coefficient, 1.548, 1.933, −0.0188, −0.0184, respectively, 95%CI, 0.047 – 3.048, 0.205 – 3.662, −0.033 – −0.0045, −0.035 – −0.021, respectively, P<0.05), but the associations between them and maximum diameter are not at all (Table).ConclusionsLower shear stress and stronger disturbed flow are reported to be risk factors for aneurysm rupture. The data by these regression analyses suggest that the surface area is a more useful risk factor for cerebral aneurysm rupture than the maximum diameter.Support or Funding InformationThis study is supported by the Grant‐in‐Aid for the Japanese National Hospital Organization Multi‐Center Clinical Research and AME‐CREST, AMED.