bluuebunny/crossref_metadata_2025_split huggingface dataset:
4β‐hydroxywithanolide E inhibits tumor necrosis factor α‐induced tissue factor expression and tumor‐associated coagulation in non‐small cell lung cancers
Authors: Hsieh Kan‐Yen , Lin Ya‐Han , Chang Fang‐Rong , Wu Chin‐Chung 
Published: June 2021

The relation between cancer and inflammation has been widely discussed, and the onset of inflammation in tumor microenvironment promotes the progression of cancer is generally recognized. Tumor necrosis factor (TNF) α, a major inflammatory cytokine, which exists in tumor microenvironment and is constitutively produced by both stromal cells and malignant cells. The implication of TNFα in tumor growth and metastasis has been reported in several literatures. In addition, TNFα causes tissue factor (TF) upregulation in several cell types, including monocyte, endothelial cells, and cancer cells that may contribute to tumor‐associated coagulopathy and cancer metastasis. 4β‐hydroxywithanolide E (4HW) is a natural compound isolated from an edible plant, Physalis peruviana. 4HW has been reported to exhibit promising anti‐cancer and anti‐inflammatory activities. In the present study, we examined the effect of 4HW on TNFα‐induced TF expression and procoagulant activity in two non‐small cell lung cancer cell lines, A549 and H1299. At the non‐cytotoxic concentrations, 4HW potently prevented TF expression caused by TNFα in both cell lines. The procoagulant activity of TNFα‐induced TF was further studied by using plasma clotting assay. The plasma clotting time in the presence of TNFα‐treated cancer cells was obviously shorter than control group, and this phenomenon was successfully reversed by pretreating with either 4HW or anti‐TF antibody. Our results suggest that 4HW might have the potential to treat tumor‐associated thrombosis and inflammation.Support or Funding InformationThis research was supported by Ministry of Science and Technology (MOST 106‐2320‐B‐037‐003‐MYB) and Kaohsiung Medical University (KMU‐TP105H03)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Natural Products as a Source to Discover Novel Drug Targets in P. falciparum
Authors: Butler Joshua H. , Zhou Bin , Yue Jian‐Min , Cassera Maria Belen 
Published: June 2021

Malaria is a deadly infectious disease that caused an estimated 216 million cases around the world in 2016 according to the World Health Organization. Plasmodium falciparum is the parasite responsible for causing the majority of malaria cases in humans. Resistance to all current frontline antimalarial therapies is a growing problem thus, there is an urgent need for new antimalarial drugs with novel targets. Natural products have a lengthy history of utility in drug discovery and remains a great source of new therapeutics. We have screened against P. falciparum in vitro cultures over 230 natural compounds isolated from various plants indigenous to the southern parts of China which have long been used in traditional Chinese medicine to treat malaria. This collection presented different degrees of antimalarial activity and toxicity toward mammalian cells. Two sesquiterpenoid dimers that exhibited potent antiplasmodial activities (<100 nM) and low toxicity to mammalian cells were selected to study their mechanism of action in more detail by inducing drug resistance in in vitro culture of P. falciparum. We were able to obtain resistant parasites against one of the selected sesquiterpenoid dimers. Clonal subcultures were subjected to next generation Illumina sequencing to identify the potential mechanism of resistance that may lead to the identification of the molecular target. In addition, a subset of similar sesquiterpenoid dimers present in this collection of natural products were screened against the resistant parasites and the analysis has outlined a starting point for structure‐activity relationship studies. We have identified a new class of promising antiplasmodial compounds due to their unique scaffold that is different from all currently known antimalarials and is likely acting through a new mechanism of action.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Montmorency tart cherry anthocyanins: dose‐dependent antioxidant activity against cholesterol oxidation
Authors: Meza Ilce Gabriela Medina , Schweiss Matthew Daniel , Barnaba Carlo 
Published: June 2021

Cardiovascular disease (CVD) is a leading cause of death world‐wide, and cholesterol oxidation (CO) and cholesterol oxidation products (COPs) are widely thought to contribute to CVD by exacerbating mechanisms that produce atherosclerotic plaques. COPs not only hold atherogenic properties, but also mutagenic, carcinogenic and inflammatory properties. More recently, it has been reported that they also play an important role in the onset and progression of neurodegenerative diseases. As promoters of inflammation, cytotoxicity, and fibrogenesis, COPs drive a cycle of inflammation that increases the steady‐state level of radical and non‐radical reactive oxygen species (ROS), which in turn amplify inflammation.Strong evidence indicates that natural products derived from plants, such as anthocyanins, poses antioxidant properties and may inhibit lipid oxidation and its metabolites. Montmorency cherries (Prunus cerasus) contain polyphenols in amounts of 3000 mg/100g dry weight, because of this they could act as a natural source of strong antioxidants. Our group has fractionated and characterized different polyphenols from Montmorency tart cherry extracts. Our preliminary studies indicate that anthocyanins have potent dose‐dependent antioxidants activity against CO suggesting that they may also be efficient at reducing CO‐induced inflammation in the CVD. We explore the antioxidant properties of Montmorency tart cherry anthocyanins against CO using in vitro systems of CVD, such as biomimetic liposomes and low‐density lipoprotein (LDL). Radical‐induced oxidation was triggered by addition of AAPH radical generator at 0.5–2 mM to a large unilamellar vesicles (LUVs) suspension under physiological conditions. Different concentrations of anthocyanins (2–100 μM) were tested over 180 minutes.Results demonstrated that anthocyanins may have a dose‐dependent antioxidant activity, specifically targeting CO. UV‐Vis spectroscopy showed that an increase in the oxidation of liposomal cholesterol when AAPH was added to the system (Control). A dramatic increase in cholesterol oxidation was observed between 180 and 240 minutes. The kinetics associated with the formation of cholesterol hydroperoxides were delayed up 180 min, with subsequent decrease on the accumulation of 7‐ketocholesterol (7‐Keto) and 25‐hydroxycholesterol (25‐OH), which are biomarkers of CO and thus the cardiovascular inflammation. We speculate that specific compounds derived from Montmorency tart cherries have the ability of to penetrate lipid membrane and scavenge ROS.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A small molecule virulence factor suppresses plant immune response
Authors: Li Bo , O'neill Erinn Marie , Mucyn Tatiana , Patteson Jon B. , Baccile Joshua , Schroeder Frank C. , Massolo Elisabetta , Dangl Jeff 
Published: June 2021

Natural products play important roles in the chemical interactions between bacteria and their host. Thousands of biosynthetic gene clusters for natural products have been identified through genomic analyses, but most of them are uncharacterized (“cryptic” gene clusters). Our goal is to identify the natural products encoded by these cryptic gene clusters and investigate their functions in bacteria‐host interaction. Using transcriptomics, we identified a cryptic gene cluster that is upregulated in Pseudomonas syringe during plant infection. This gene cluster is widely distributed in plant pathogens that cause worldwide crop damage. To identify the natural product encoded by this gene cluster, we develop an approach that combines heterologous expression and metabolomics with in vitro reconstitution of enzyme activity. We identify a novel natural product, phevamine, which harbors a distinct chemical structure that partly resembles spider and wasp toxins. We discover that phevamine is a virulence factor that suppresses plant immune response. The biosynthesis of phevamine involves ATP‐Grasp type amide ligases. Our work identifies a new virulence factor that pathogens use to invade plant defense. Our work also highlights amide ligases as an important and growing class of biosynthetic enzymes for natural products.Support or Funding InformationThis work was supported by Rita Allen Foundation (B.L.), National Institutes of Health (R00 GM099904, B.L.), NSF‐IOS‐1343020 INSPIRE Track 2 (J.D.), and DOE‐BER‐0000217779 (J.D.).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Engineering A Hybrid FRET Biosensor to Study Proteolytic Activities of MT1‐MMP
Authors: Limsakul Praopim , Wang Yingxiao 
Published: June 2021

The genetically‐encoded biosensors based on Förster resonance energy transfer (FRET) are extensively used for real‐time monitoring of biological processes including enzymatic activities in living cells with high spatiotemporal resolutions. However, the development of a membrane‐targeted FRET biosensor remains a challenge partly due to the poor efficiency of transportation and integration of fusion proteins at the plasma membrane, which results in high background signals. To overcome this deficiency, we introduced an alternative strategy that allows the direct assembly of FRET biosensors in situ at the cell surface by integrating an enhanced cyan fluorescence protein (ECFP; a donor) and a fluorescent dye, R‐Phycoerythrin (R‐PE; an acceptor), for FRET action. We utilized the directed evolution technology and sequence‐function analysis to engineer a monobody into an efficient R‐PE binder, PEbody, to directly capture R‐PE. Our hybrid FRET biosensor was used to monitor the enzymatic activity of the membrane type 1 metalloproteinase (MT1‐MMP) on the extracellular surface of live cancer cells. The results revealed a cytoskeleton‐dependent distribution of MT1‐MMP activities at different cell‐cell contacts, with high and low activities at loose and stable cell‐cell contacts, respectively. Therefore, integrating the directed evolution approach and rational design can be employed to systematically design novel FRET biosensors capable of monitoring the molecular dynamics at the surface and other subcellular regions in living cells.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Phospholipids as Indicators of Castration Resistant Prostate Cancer
Authors: Ingram Lishann M. , Manosoura Maryam , Pati Sumitra , Cummings Brian S. 
Published: June 2021

The association of circulating lipids with clinical outcomes of metastatic castration‐resistant prostate cancer (CRPC) is not fully understood. Interestingly, recent studies demonstrated a correlation between plasma lipids and CRPC prognosis, resulting in a proposed prognostic four‐lipid signature of (ceramide, sphingomyelin, phosphatidylcholine (PC) and phosphatidylserine (PS)). While the increase in these lipids correlated to decreased survival, the mechanisms mediating alterations in these lipids, and the correlation to resistance to treatment are not known. We addressed this gap‐in‐knowledge using in vitro models of non‐cancerous, hormone sensitive and CRPC cell lines and quantitative lipidomic analysis. An initial untargeted approach assessing both cells and media indicated that ceramides, sterols and glycerophospholipids had higher relative abundance in CRPC cells (PC‐3 and DU145), as well as hormone sensitive cells (LNCaP), as compared to a non‐cancer cell model (PNT2). Similar data was seen in media. A targeted approach focusing on lipid species shown to correlate to survival in recent publications showed significantly higher levels of 36:2 phosphatidylserine (PS) in all prostate cancer cell lines tested, and lower levels of 36:2 PC in the media of these same cells. Thus, both untargeted and targeted analysis of prostate cancer cells showed significant differences in lipidomic profiles as compared to non‐cancer prostate cells. These data suggest that the lipidomic profiles of prostate cancer cells lines mirror lipidomic profiles in the plasma of prostate cancer patients. As such, these cells may prove useful for understanding the molecular mechanism mediating lipid changes in prostate cancer patients and yield insight into the role of lipids in mediating CRPC.Support or Funding InformationNational Institute of Biomedical Imaging and Bioengineering NIBIB (EB0160100)Department of Defense Prostate Cancer Research Program Idea Development Award (PC150431 GRANT11996600)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Defects in TM6SF2 Impairs Lipidation of Nascent VLDL and Leads to Accumulation of Liver Fat
Authors: Martin Sarah A. , Smagris Eriks , Cohen Jonathan C. , Hobbs Helen H. 
Published: June 2021

Non‐alcoholic fatty liver disease (NAFLD) is a burgeoning health problem in Western countries. Population‐based surveys indicate that one out of every three adults in the United States has excess liver fat (hepatic steatosis), the first stage of fatty liver disease. Despite the clinical importance of NAFLD, major questions remain regarding its etiology. As a step towards elucidating the pathophysiology of this disorder, we screened individuals in a population‐based study, the Dallas Heart Study (DHS) for DNA sequence variations associated with increased liver fat content. This analysis revealed that a common variant (E167K) in transmembrane 6 superfamily 2 (TM6SF2), a gene of unknown function, was associated with increased hepatic triglyceride (TG) content, reduced plasma lipid levels, and increased levels of circulating liver enzymes. TM6SF2 is expressed at highest levels in the small intestine, liver and kidney. Immunolocalization and biochemical studies localized the protein to the ER and the Golgi. To determine why individuals with the risk variant develop hepatic steatosis, we developed mice expressing no TM6SF2 (Tm6sf2−/−, KO). The Tm6sf2−/− mice have elevated levels of neutral lipids, including TG and cholesteryl esters (CE) in the liver and elevated levels of liver enzymes in the circulation, consistent with liver inflammation. At steady state the Tm6sf2−/− mice have decreased plasma levels of cholesterol and very low density lipoprotein (VLDL)‐TG. Treatment with Triton WR1339 to inhibit peripheral lipases was associated with a non‐specific and dramatic reduction in all of the VLDL associated lipid molecular species, but not apolipoprotein (Apo)‐B100. In addition, it was found that de novo synthesis of radioactive and stable labeled lipids was marginally changed in the liver, but secretion of the newly synthesized labelled lipids was dramatically decreased in the Tm6sf2−/− mouse. As expected, given these observations, VLDL was reduced in size in the KO mice. Thus, TM6SF2 plays a key role in the expansion of the nascent particle, but not in the secretion of nascent VLDL, thereby causes steatosis.Support or Funding InformationFunding was provided by the NIH through research grants R01‐DK090066 and F32‐DK112529.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Metabolomic profiling associated with deployment‐related stressors in Army personnel
Authors: Gautam Aarti , Donohue Duncan , Abu‐Amara Duna , Hoke Allison , Genfi Afia , Blessing Esther , Hammamieh Rasha , Marmar Charles , Jett Marti 
Published: June 2021

Pre‐deployment identification of risk and resilience factors is crucial in developing targets to reduce or prevent posttraumatic stress symptoms in military personnel. The Fort Campbell Cohort study was designed to assess pre‐deployment biological and behavioral markers and build predictive models to identify risk and resistance for posttraumatic stress disorder (PTSD) following deployment. The aim of the current study was to use high‐resolution metabolomics profiling to identify metabolic pathways and networks associated with PTSD checklist score differences and changes across multiple pre and post‐deployment time points. In addition, we have investigated how deployment affects metabolomics profile changes within individuals independent of PTSD score changes. Untargeted metabolomics profiling of plasma has been completed using both liquid and gas chromatography with mass spectrometry. Our preliminary data indicates that deployment has complex metabolic effects that must be considered in evaluation of deployment‐associated exposures in military personnel. Our in depth‐data analysis may further elucidate pathways, novel compounds, and biomarkers associated with PTSD in its relation to military deployment. These findings may have clinical implications for understanding the pathogenesis of PTSD and provide insights to avert chronic PTSD.Disclaimers: Research was conducted in compliance with all Federal requirements. The views expressed are those of the authors and do not constitute endorsement by the U.S. ArmySupport or Funding InformationFunding support from Military Operational Medicine Research Program, MOMRP is highly acknowledged.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Distinct Transcriptional Profile Activated by SCYE1 Through JAK‐mediated STAT3 in Partially Activated Macrophages
Authors: Lee Daniel , Schwarz Margaret 
Published: June 2021

Small inducible cytokine subfamily E, member 1, SCYE1 (also known as EMAP II) is a secreted factor that is locally expressed near sites of injury, labeled as a ‘find‐me’ signal, and recruits macrophages and neutrophils. Our previous studies identified that macrophages sustained protein expression of SCYE1. The transcriptional events that occur within the recruited macrophages in response to SCYE1 are unknown. The objective was to identify the transcription factor activated in partially activated thioglycollate macrophages upon exposure to SCYE1.Partially activated murine thioglycollate‐elicited macrophages (TEPMs) were exposed to SCYE1. Following 2 hours of exposure to either vehicle or SCYE1, fractionated nuclear proteins from TEPMs were used for a luminescence based transcription factor activation screen. The acute SCYE1‐mediated transcription factor activation was assessed by immunoblotting and gene expression by quantitative PCR. Secreted cytokines were measured in supernatants of TEPMs exposed to either vehicle or SCYE1 while pre‐treated with +/− JAK1/2 inhibitor, Ruxolitinib.In this study, we identify STAT3 (Signal Transducer and Activator of Transcription 3) as the most activated transcription factor mediated by SCYE1 in a model of partially activated, recruited thioglycollate‐elicited peritoneal macrophages. The acute and dose‐dependent phosphorylation of STAT3 (Y705) occurs between 5 and 15 minutes in response to SCYE1. Its activation is SCYE1‐specific and could be prevented by inhibition of Janus Associated Kinase (JAK) 1/2.We profiled not only anti‐ and pro‐inflammatory genes but also that of STAT3 target genes upon exposure to SCYE1. STAT3 target genes Arg1 and Socs3, commonly activated in anti‐inflammatory macrophages, are maximal by 1 to 2 hours of exposure to SCYE1. Yet Il10 expression decreased by 1 hour suggesting that any persisting STAT3 activation was not due to Il10. There was also rapid increase of chemokine STAT3 target genes Ccl5 along with other genes activated that are typically found in pro‐inflammatory macrophages.Unlike activated transcription, neither secreted IL‐6 nor IL‐10 was elevated by SCYE1. Consistent with a JAK‐STAT signature, both IL‐1a and G‐CSF secretion by SCYE1 was elevated and could be prevented by JAK inhibition.Our results identify a rapid SCYE1‐mediated STAT3 activation that coincides with both pro‐ and anti‐inflammatory gene expression in partially activated macrophages.Support or Funding InformationFunding: This publication was made possible in part by 5R01HL114977 (MAS) from NIH and the Lilly Endowment, Inc. Physician Scientist Initiative (MAS).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Perturbation of Homocysteine Homeostasis Leads to Cellular Epigenetic Modification And Endothelial Injury in Acute Myocardial Infarction
Authors: Chang Christopher , Yang Tzu‐Ching , Lu Shao‐Chun , Lee Ming‐Syue , Chang Po‐Yuan 
Published: June 2021

Homocysteine is a highly reactive, sulfur‐containing amino acid formed as a byproduct of the metabolism of the essential amino acid methionine. Homocysteine used to be thought of as a risk factor for atherosclerosis, but emerging data have suggested that S‐adenosylhomocysteine (SAH), an intermediate metabolite of homocysteine, might be a better indicator of vascular diseases. In this study, we investigated the epigenetic role and clinical significance of SAH‐homocysteine homeostasis in acute myocardial infarction. A cell culture system was established to assess the interplay between SAH and homocysteine using human coronary artery endothelial cells (ECs). Homocysteine alone did not affect EC survival up to 500 uM; however, in the presence of adenosine/EHNA, homocysteine (25–500 uM) dose‐dependently increased intracellular SAH and promoted TUNEL‐positive apoptotic cell death. There was a concomitant downregulation of fibroblast growth factor 2 (FGF2), a critical cytokine for EC survival, through extensive DNA methylation at CpG dinucleotides in the FGF2 gene promoter. These pro‐apoptotic effects in ECs were markedly attenuated when adenosine/EHNA was removed from the culture medium. In patients with acute ST‐elevation myocardial infarction (STEMI), the plasma SAH levels quantitated by liquid chromatography coupled to tandem mass spectrometry (LC‐MS/MS) were significantly higher than those in healthy controls. In contrast, the plasma homocysteine concentrations were similar between STEMI and healthy subjects. Moreover, STEMI patients who had an obvious clinical improvement also demonstrated a significant decrease in their plasma SAH levels at subsequent follow‐up. Our data conclude that the perturbation of SAH‐homocysteine homeostasis in ECs leads to cellular epigenetic modification and endothelial injury in acute myocardial infarction. Elevation of plasma SAH but not homocysteine in STEMI patients further supports the notion that SAH is a better indicator of vascular diseases than homocysteine.Support or Funding InformationThis work was supported by grants NSC101‐2320‐B‐002‐026 from the National Science Council and MOST‐106‐2314‐B‐002 ‐156 ‐MY2 (Dr. Po‐Yuan Chang) from the Ministry of Science and Technology, Taipei, Taiwan.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.