bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Pulmonary Valve Anatomy and Morphometry
Authors: Holda Jakub , Holda Mateusz Krystian , Tyrak Kamil , Klimek‐Piotrowska Wieslawa 
Published: June 2021

BackgroundThere is a strong clinical need of accurate pulmonary valve anatomy knowledge. Thus, the aim of this study is to present detailed morphometric description of the human pulmonary valve.MethodsA total of 200 autopsied human hearts from healthy donors (26.0% females) were investigated. We have measured several pulmonary valve parameters and noted the presence of calcifications, semilunar valve nodules and leaflets' fenestrations. This study has been conducted in conformance with the Basic Principles of the Declaration of Helsinki and their expansion.ResultsAmong study material 199 pulmonary valves were tricuspid and only one was bicuspid. The mean diameter of the pulmonary valve was 20.0±4.1 mm in the frontal plane and 18.0±4.0 mm in the sagittal plane. The mean distance from the free margin to the attachment line of the leaflet (leaflet height) in its central part was 15.6±3.4 mm for left, 15.1±3.1 mm for anterior and 15.4±2.8 mm for right leaflet. The mean distance from the bottom of the pulmonary sinus to the sino‐tubular junction for the left, anterior and right sinuses were: 19.0±3.8 mm, 19.1±3.7 mm and 18.4±4.1 mm, respectively. The distance from the free margin of the leaflet to the pulmonary sinus wall (in closed valve) was 15.9±2.8 mm for left, 15.5±2.9 mm for anterior and 16.1±2.8 mm for right leaflet. No statistically significant differences in size of leaflets and sinuses were found (p>0.05). Nodule of semilunar valve in the left, anterior and right leaflet was observed in 71.0%, 85.5% and 85.5% of hearts respectively. In 52.5% of cases nodules were presented in all three leaflets. In 22.0% of cases fenestrations of at least one leaflet were seen. The myocardial sleeve from the right ventricle tissue was situated higher than the bottom of the right pulmonary sinus in 87.5% of cases. The mean height of the myocardial sleeve extension onto right pulmonary sinus was 3.4±1.5 mm. Calcifications of pulmonary valve were observed only in 2.0% of cases.ConclusionThe pulmonary valve has almost always three leaflets, which are of comparable size (p>0.05). The myocardial sleeve of right ventricle tissue entering the pulmonary valve sinus is present in 87.5% of cases and should be taken into consideration during pulmonary valve surgical procedures. The presence of leaflets' fenestrations may be responsible for valvular leak.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
mTORC1 controls GSK3β nuclear localization
Authors: Antonescu Costin N. , Bautista Stephen , Boras Ivan , Vissa Adriao , Mecica Noa , Yip Christopher M. , Kim Peter 
Published: June 2021

Glycogen synthase kinase 3β (GSK3β) is a serine/threonine kinase that regulates numerous cellular processes such as apoptosis, self‐renewal, growth, and metabolism. GSK3β phosphorylates over 100 substrates ranging from cytosolic to nuclear substrate targets, including c‐myc, a key ongogenic transcription factor, leading to control of c‐myc stability. GSK3β is itself regulated by phosphorylation on S9 by signals such as Akt (activated within the phosphatidylinositol‐3kinase, PI3K, signaling pathway), resulting in reduced GSK3β kinase activity. Given the wide range of substrates and the incomplete regulation of GSK3β by S9 phosphorylation, it is likely that other mechanisms gate GSK3β function.GSK3β may be localized within different cellular compartments such early endosomes, the nucleus, and in multivesicular bodies. However, how GSK3β localization is regulated, and how this may control GSK3β function is poorly understood, which we have examined here. Specifically, we examined how mTORC1, a key cellular sensor of mitogenic and metabolic signals, controls GSK3β localization and function. Using pharmacological inhibitors of mTORC1 and GSK3β, as well as siRNA gene silencing approaches, we uncovered that GSK3β‐mediated c‐myc degradation is negatively regulated by mTORC1. Using fluorescence microscopy approaches, we find that inhibition of PI3K‐Akt‐mTOR signaling axis, and metabolic insufficiency (e.g. amino acid deficiency), elicit GSK3β nuclear translocation from the cytosol, suggesting that mTORC1 mediates GSK3β nucleocytoplasmic shuttling. Consistent with regulation of GSK3β by mTORC1, we also uncovered that GSK3β localizes to LAMP1 positive late endosome/lysosomes, and perturbations of late endosome/lysosome membrane traffic impacted nuclear localization of GSK3β. This suggests that late endosomes/lysosomal compartments serve as organizing platforms, integrating mitogenic and metabolic signals via mTORC1 to elicit GSK3β cytosolic retention, thus impacting access of GSK3β to nuclear targets and controlling c‐myc degradation. Understanding how mTORC1 dependent signals regulate GSK3b subcellular localization to control GSK3β function may give useful insight to the development of drugs and therapies targeting GSK3β for the treatment of cancer cell growth and survival.Support or Funding InformationThis work is supported by a Discovery Grant from the Natural Science and Engineering Research Council (of Canada), an Early Researcher Award from the Ontario Ministry of Research, Innovation and Science, and a New Investigator Award from the Canadian Institutes of Health Research to C.N.A.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Localized immune transcriptional response to intratumorally administrated TLR7 agonist conjugates
Authors: Echeagaray‐Bredesen Oscar E , Vallez Nick , Mendez Natalie , Huang Ching‐Hsin , Gude Natalie , Kummel Andrew 
Published: June 2021

Induced or circumstantial immunosuppression present opportunities for cancer development, and progression. Recently, multiple therapies have been developed to harness the immune system's ability to selectively eradicate cancer cells. These approaches have primarily targeted features of the adaptive immune response such as checkpoint blockade and vaccination of tumorigenic peptides. Toll‐Like receptors (TLRs) are innate immune receptors that recognize molecules broadly shared by various pathogens. TLR pathways are increasingly targeted in cancer immunotherapy approaches seeking to overcome immune‐evasion by cancer cells. Here, we hypothesize that activation of TLR7, an endosomal receptor which recognizes pathogen associated nucleic acids, can mitigate tumor immunosuppression. TLR7 agonist (TLR7‐A) conjugated to silica nanoparticles will be used as an adjuvant in a mouse colon adenocarcinoma (CT26 cell line derived) in vivo tumor model. Transcriptional targets of adaptive immune cells (CD3, CD4, CD8, Granzyme B and Perforin), antigen presenting cells (MHCII, CD80, CD86 and CD40) and inflammatory cytokine response (TNFα, IL‐6, IP‐10, MCP1, MIP1α and IFNγ) were measured by qPCR in tumor tissue following 21 days of treatment. We found that treatment with TLR7‐A unconjugated and conjugated increased localized transcript levels of inflammatory cytokines and chemokines: IL‐6, IP‐10, MCP1, MIP1α and IFNγ. This project will provide valuable insights on the distinct transcriptional immune interplay in response to TLR7‐A treatment (localized vs systemic), support the decision‐making process of subsequent experimental approaches and evaluate TLR7‐A conjugate's potential for treatment.Support or Funding InformationThis research was supported by the National Cancer Institute of the National Institutes of Health under award numbers:U54CA132384 & U54CA132379This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mast Cell Numbers In Rat Livers In An Acute Model Of Fat Embolism Are Reduced By Losartan But Not By Captopril
Authors: Siddiqi Ahsan , Hamdan Hana , Siddiqi Saba , Pour Mohammad , Randall Zachary , Leupold Brad , Arif Dauod , Poisner Alan , Hamidpour Soheila , Molteni Agostino 
Published: June 2021

IntroductionFat embolism (FE) induced by IV injection of triolein (T) causes vasculitis, septal inflammation and fibrosis in rat lungs within 48 hrs (1). The damage is mitigated by two drugs acting on the renin‐angiotensin system (RAS): captopril and losartan (2). In the same organ, mast cell numbers are reduced in the lungs by losartan but not by captopril (3). This study examines whether captopril and losartan also modify the mast cell response in the liver.MethodsSprague‐Dawley rats (280–300g) were treated with 0.2ml iv (T) n=18 or saline n=7. One hour later 0.2ml of saline (n=6) ip, captopril (150 mg/kg n=6) or losartan (10mg/kg n=6) were given. Saline treated rats received ip, one hour later, the same does of saline. 48 hours later all the rats were necropsied after isoflurane anesthesia, lungs fixed in 10% formalin and H&E stained for morphology and CD117 for mast cells. Two pathologists unaware of the slides identity took at random 10 photographs of all the slides at 400× and separately counted the mast cells.ResultsAs previously reported, 48 hours after triolein the lungs showed severe histopathological damage that was prevented by both RAS drugs and was associated with a high number of mast cells after captopril treatment but not by losartan (3). Similar results were observed in the liver where the mast cells after 48 hours of triolein treatment were statistically significantly increased (p=0.019) and even more when triolein treatment was associated with captopril (p=0.009). Mast cells numbers in the liver of losartan treated rats was however similar to that of controls and no statistically significant difference was observed. The overwhelming majority of mast cells were observed within the portal tract approximating 78%. The remaining were spread out within the hepatic parenchyma.ConclusionMast cells may be involved in the pathological damage found after FE as suggested by their increase in number in the lungs and the liver after T and their reduction during protective treatment with losartan. However, the lack of effect of captopril on mast cell number at this acute period both in lungs and the liver may suggest that some other pathway may be involved in mast cell recruitment such as angiotensin 1–7.Support or Funding InformationSupported by the Catherine T. Geldmacher Foundation, St. Louis MO.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cultivation establishment and antiproliferative activity from Asclepias subulata, for the development of a phytopharmaceutical
Authors: Armenta Hector Enrique Bustamante , Elías Jesús López , Rodríguez Julio César , Valenzuela Luisa Alondra Rascón , Zepeda Ramón Enrique Robles 
Published: June 2021

Cancer is one of the leading causes of death in the world, this disease is commonly treated with non‐specific therapies, an alternative is the use of phytopharmaceuticals. Asclepias subulata is a medicinal plant, whose extracts report a high antiproliferative activity and selectivity on human carcinogenic cell lines. For the eventual development of a phytopharmaceutical from A. subulata, it is necessary to establish the conditions for the domesticated cultivation of this wild species. The aim of this work was the cultivation establishment of A. subulata and determine the antiproliferative activity of the plant under water stress. The seeds were collected and germinated in a greenhouse. Once germinated, were transplanted to the field, under four different conditions: Drought stress (DS), Drought stress with gel (DG), Irrigation (I) and Irrigation with gel (IG). Monthly for a year, aerial biomass and water stress were evaluated for each treatment. In addition, ethanolic extracts were obtained to determine the profile of cardenolides (HPLC) and antiproliferative activity on the cell lines A549 and ARPE‐19. The results obtained from aerial biomass in April‐December were, for DH, DG, I and IG were 75.9 ± 27.1, 83.1 ± 31.1, 323.8 ± 139.5 and 335.4 ± 93.0 g / plant respectively. The IG treatment was the one with more production of aerial biomass. The ethanolic extracts showed strong antiproliferative activity with values of IC50 <3.5 μg / mL, on the A549 cell line. On the other hand the ARPE‐19 cell line show IC50 >200 μg / mL, all these data can indicate the selectivity of the ethanolic extracts towards human cancer cell lines. We did not significate differences between treatments or season in the antiproliferative activity. Additional studies are required to determine other factors could be influencing the production of cardenolides.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Assessment of Novel Vitamin D Receptor Antagonists that Mediate Suppression of Vitamin D Signaling
Authors: Staniszewski Lech J.P. , Shahani Pritika , Heck Michael , Hasan Dania S. , Wagner Carl , Jurutka Peter W. 
Published: June 2021

Antagonists are chemical compounds that bind to receptors without eliciting activation of signaling, therefore the biochemical response to the receptor ligand can be blocked or significantly decreased. Receptor antagonists display varying levels of binding affinity but do not produce the agonist‐mediated response upon binding. The vitamin D receptor (VDR) binds with high affinity to its endocrine agonist, 1,25‐dihydroxyvitamin D3 (1,25D) to regulate the expression of a suite of genes in target tissues including kidney, intestine, and bone to control biological processes such as calcium and phosphate bone mineral homeostasis. In conditions of VDR hyperactivity, excess vitamin D production, or significantly increased intake via diet or supplementation, there are risks associated with potential resultant hypercalcemia including kidney stones, GI dysfunction, behavioral changes such as depression, and cardiac problems. In the present study, we synthesized novel VDR antagonists to create synthetic ligands that have high affinity for VDR, and consequently inhibit the activation of 1,25D‐VDR regulated genes that lead to hypercalcemia. It is well established that agonist‐bound VDR forms a functional DNA‐binding heterodimer with the retinoid X receptor (RXR) that associates with vitamin D responsive elements (VDREs) to either induce or repress transcription. Therefore, we exploited this agonist‐dependent VDR‐RXR heterodimerization pathway as an initial screening assay to evaluate the efficacy of our putative VDR antagonists. The initial test employed the mammalian two‐hybrid assay (M2H), which can assess if the novel antagonists compete with 1,25D and bind to VDR to prevent the formation of a heterodimer with RXR, a prerequisite step in VDR signaling. A second assay utilized VDRE‐linked luciferase reporter plasmids to measure antagonist activity in the more natural environment of the VDRE DNA platform. This assay was followed by a third screen to determine the specificity of antagonist binding to VDR versus other closely related receptors in the nuclear receptor superfamily. Results from these assays revealed that two novel analogs bind as bona fide antagonists to the VDR, albeit with different affinity. Our findings also suggest that additional chemical modification of antagonists may result in analogs with enhanced ability to suppress 1,25D‐VDR activation. Thus, this study further broadens our understanding of VDR‐ligand binding interactions, and may facilitate the development of novel therapeutic compounds with the potential for clinical applications in treating VDR‐directed hypercalcemia.Support or Funding InformationSchool of Mathematical and Natural Sciences at Arizona State UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Discovery of a Plant‐derived Cell‐penetrating Proteasome Inhibitor
Authors: Kam Antony , Loo Shining , Tam James P 
Published: June 2021

Plant‐derived cysteine‐rich peptides (CRPs) are known to play a role in plant defence against fungi and insects [1]. An increasing body of evidence, however, showed that certain CRPs display cell‐penetrating functions, and could be exploited for therapeutics targeting intracellular proteins [2]. Hibiscus sabdariffa, commonly known as roselle, is a shrub cultivated in South‐east Asia. We previously reported the discovery of CRPs from roselle, which we designated as roseltides (rT1‐rT7) [3]. Roseltides possess an evolutionary conserved six‐cysteine motif typical of hevein‐like peptides, but without a chitin‐binding domain. Roseltide rT1 is a human neutrophil elastase inhibitor, which in part, correlates to the ethnomedicinal use of roselle. Here we report that roseltide rT7, the second most abundant CRP in roselle, is a cell‐penetrating peptide and a proteasome inhibitor. To prepare the synthetic roseltide rT7, we used a stepwise solid‐phase peptide synthesis, followed by oxidative folding, to afford a native‐folded rT7. We showed that the synthetic and native roseltide rT7 are indistinguishable using reversed‐phase high‐performance liquid chromatography and nuclear magnetic resonance spectroscopy. To gain insight into its cell‐penetrating properties, we selectively labeled roseltide rT7 at its N‐terminus with a fluorescent probe (TAMRA‐rT7). Using flow cytometry and live‐cell confocal microscopy, we showed that TAMRA‐rT7 is cell membrane permeable via the endocytotic pathway. Roseltide rT7 was found to inhibit the chymotrypsin‐like activities of human 20S‐proteasome. Taken together, this study highlights the discovery of roseltide rT7 as the first plant‐derived cell‐penetrating proteasome inhibitor that can be potentially useful in the development of therapeutic biologics for cancer, inflammatory, and heart diseases.Support or Funding InformationThis research was supported in part by Nanyang Technological University Internal Funding ‐ Synzymes and Natural Products (SYNC) and the AcRF Tier 3 funding (MOE2016‐T3‐1‐003).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differentiation of Sickle Cell Zygosity Utilizing a Sodium Metabisulfite Method
Authors: Le Austin , Uppugunduri Manoja , Korducki John , Randolph Tim 
Published: June 2021

Hemoglobinopathies are the most prevalent monogenetic disorder worldwide, and sickle cell disease is the most prevalent and severe structural hemoglobinopathy. Globally, over 85% of the sickle cell gene penetrance occurs in sub‐Saharan Africa (64.4%) and Arab‐India (22.7%) which contains the poorest countries on earth. Diagnosing sickle cell disease remains problematic in these populations where poverty severely limits clinic budgets, country infrastructure, and healthcare education making the use of modern diagnostic methods impractical. By modifying an established sodium metabisulfite method to detect sickle cells, we developed a simple, rapid, cost‐effective, microscopic method to distinguish the homozygous genotype (SS) seen in sickle cell disease (SCD) from the heterozygous genotype (AS) seen in sickle cell trait (SCT). We hypothesized that the two zygosities can be differentiated by observing the number, intensity, and rate of sickle cell formation over time. A solution of 2% sodium metabisulfite was prepared and 15 μL was mixed with 15 uL of patient blood and 5uL of the mixture was transferred to a microscope slide for evaluation using 100x oil immersion brightfield microscopy. Sickle cells were counted in one hour intervals for three hours and judged for sickling intensity on a 0–4+ Likert Scale. It was determined that 4+ sickle cells at 3 hours incubation optimally differentiated SS from AS genotypes. SS samples had an average of 135, 4+ sickle cells (N=10) compared to 12, 4+ sickle cells in the sickle cell trait samples (N=5) at three hours. Although more data are being collected, this inexpensive method can potentially be used to diagnose SCD and SCT, benefiting countries where there are limited resources.Support or Funding InformationAL is supported by DeNardo Education and Research Foundation Fellowship.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of Reactive Oxygen Species in Crotalus atrox Venom‐induced Cell Death
Authors: Brown Lindsay 
Published: June 2021

Venomous snake bites impact humans all around the world. Anti‐venom treatments mitigate systemic effects such as vascular hemorrhage, platelet aggregation inhibition, and the activation of inflammatory mediators. However, hemorrhagic snake venom also causes a loss of cellular adhesion to extracellular matrix components resulting in massive local tissue damage. To better understand the mechanism in which venom induces local tissue damage human embryonic kidney cells (HEKS) were grown on PEI then stimulated with 500μg/ml Crotalus atrox venom for 4 and 10 hours. Alamar Blue assays were used to measure cell viability and results suggest a 14.5±8.6% (p<0.05) and 58.95±10.7% (p<0.05) reduction in cell viability at 4 and 10 hours, respectively. Cells stimulated with 500μg/ml venom for 10 hours stained 97.6±2.2% (p<0.05) positive for Trypan blue, suggesting the venom reduces membrane integrity. HEK cells stimulated with 500μg/ml venom for 10 hours in the presence of 200 units PEG‐catalase increased viability by 44.1±5.1% (p>0.001) compared to cells stimulated with venom alone. 2′,7′‐Dichlorofluorescin‐diacetate (DCF‐DA) was used to quantify reactive oxygen species during venom stimulation. HEK cells stimulated with 50μg/ml Crotalus atrox venom for 4 hours returned 1.3×106±6.5×104 relative fluorescent units (RFU) compared to the 3.3×103±1.9×102 for non‐stimulated controls (p<0.05), a 433‐fold increase suggesting reactive oxygen species such as hydrogen peroxide play a critical role in Crotalus atrox venom induced cell death.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Serum Lipoproteins Regulate Hypoxia‐Inducible Factors Under Normoxia
Authors: Espenshade Peter , Shao Wei , Hwang Jiwon 
Published: June 2021

Hypoxia‐inducible factor (HIF) transcription factors are central regulators of oxygen homeostasis, controlling energy utilization, erythrogenesis, angiogenesis and cancer metabolism. Oxygen negatively regulates HIF activity by promoting HIFα prolyl hydroxylation and proteasome‐dependent degradation. Using human pancreatic adenocarcinoma cells, we demonstrate that serum lipoproteins are an independent regulator of HIF under normoxia. Decreasing extracellular lipid supply inhibited HIF prolyl hydroxylation, leading to accumulation of HIFα comparable to hypoxia with activation of downstream target genes. Addition of unsaturated fatty acids suppressed this signal, which required an intact mitochondrial respiratory chain. A survey of different cell lines showed that this signaling pathway functions in a wide range of tissues. These studies show that lipoprotein and oxygen supply are distinct signals integrated to control HIF activity and identify serum lipids as a potential modulator of HIF pathway activity.Support or Funding InformationNational Institutes of Health, HL077588 and GM126088This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.