bluuebunny/crossref_metadata_2025_split huggingface dataset:
Probing the structure, dynamics and regulation of lipid signalling enzymes and their role in human disease
Authors: Burke John 
Published: June 2021

Lipid phosphoinositides play fundamental roles in virtually all pathways that control a cell's decision to grow, move, divide, and die. Because of this, kinases that phosphorylate phosphoinositide lipids are critically involved in myriad essential functions including growth, development, and membrane trafficking. The mis‐regulation of the activity of phosphoinositide kinases is critical in human diseases, including cancer, primary immunodeficiencies, developmental disorders, and inflammation. Phosphoinositide kinases also play critical roles in mediating bacterial and viral infection, including many potent human pathogens. Inhibitors of parasite phosphoinositide kinases are in development as therapies for both malaria and cryptosporidiosis. Therefore, understanding how phosphoinositide kinases are regulated has implications for the treatment of many devastating human diseases.My research is focused on understanding the molecular basis for regulation of phosphoinositide kinases, and how they are involved in human disease. We have used a synergy of biophysical approaches involving X‐ray crystallography, hydrogen deuterium exchange mass spectrometry, Cryo‐EM, and biochemical assays to study the structure, dynamics, and molecular interactions of these critical signalling enzymes. I will discuss our work on studying the molecular mechanism of how mutations in phosphoinositide 3‐kinases mediate primary immunodeficiencies and cancer, and how viruses manipulate phosphatidylinositol 4‐kinase protein interaction networks to mediate infection. Overall this work is important in understanding how mis‐regulation of phosphoinositide lipid signals mediate disease. We will also discuss how structural information on these enzymes is being leveraged to generate novel small molecule inhibitors of these enzymes as potential therapeutics for cancer, viral infection, and malaria.Support or Funding InformationJ.E.B. is supported by the Canadian Institute of Health Research (new investigator grant and CIHR open operating grant FRN 142393), a discovery research grant from the Natural Sciences and Engineering Research Council of Canada (NSERC‐2014‐05218), and a Cancer Research Society operating grant (CRS‐22641).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Talk title Bridging the gap between RNA editing and modification: A 10‐year solution to a 25‐year problem
Authors: Alfonzo Juan D. 
Published: June 2021

All types of nucleic acids in cells undergo naturally occurring chemical modifications, including DNA, rRNA, mRNA, snRNA, and most prominently tRNA. Over 100 different modifications have been described and every position in the purine and pyrimidine bases has been seen modified; the sugar is often also modified. Despite recent progress, the mechanism for the synthesis of most modifications is not fully understood partly due to difficulty in reconstituting modification enzyme activity in vitro. Cytosine to uridine (C to U) editing of tRNAs in eukaryotes was first discovered over 20 years ago, yet due to the lack of an in vitro assay, the editing enzyme is still at large and the editing mechanism remains unsolved. In this work we show that cytosine 32 in the anticodon loop of tRNAThr is methylated to 3‐methylcytosine (m3C) as a pre‐requisite for C to U deamination. Formation of m3C in vitro requires the presence of both TbTRM140 (the m3C methyltransferase) and the TbADAT2/3 (the A to I editing deaminase). Once m3C is formed the same set of enzymes are required for further deamination from m3C to form 3‐methyluridine (m3U). In our previous work, we also showed that TbADAT2/3 was a highly mutagenic enzyme randomly deaminating DNA. Now we provide evidence that when co‐expressed with the methylase its mutagenic ability is kept in check via complex formation with TRM140. This helps explain how T. brucei escapes “wholesale deamination” of its genome while harboring both enzymes in its nuclear compartment. This observation has implications for the control of another mutagenic deaminase, human AID, and provides a rationale for its regulation.Support or Funding InformationNIH GM084065This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Telomerase Expression in Myeloid Cells is Essential for Pulmonary Fibrosis
Authors: Liu Tianju , De Los Santos Francina Gonzalez , Zhao Yuyue , Wu Zhe , Phan Sem H. 
Published: June 2021

Shortened telomeres and mutations in telomerase reverse transcriptase (TERT) or its RNA component (TERC) have been reported in some patients with fibrotic interstitial lung disease, such as idiopathic pulmonary fibrosis (IPF). However telomerase is induced in lung fibroblasts from such patients without evidence of telomere shortening, and selective TERT deficiency in mesenchymal cells significantly reduced pulmonary fibrosis in an animal model. Moreover TERT in the bone marrow (BM) is essential for fibrosis in the same model. However, the precise cellular source of TERT in the BM is unknown. The objective of this study was to elucidate further the identity and the role of the TERT‐expressing BM‐derived cells in the injured lung by evaluating the effects of selective TERT deficiency in myeloid (LysM+ cells) on bleomycin (BLM) induced pulmonary fibrosis. The myeloid cell specific TERT knockout mice (LysM‐TERT KO) were created by crossing floxed TERT mice with LysM‐Cre mice. The results showed that BLM induced significant TERT expression in BM‐derived CD45+ cells in lungs of control (WT) mice, which was not observed in the LysM‐TERT KO mice. This was also associated with the loss of TERT expression in whole lung tissue in LysM‐TERT KO mice while TERT expression in the BM was reduced by >80%. Interestingly, BLM‐induced TERT expression in lung CD45− cells in WT mice was also abolished by myeloid cell TERT deficiency in LysM‐TERT KO mice. The myeloid cell specific TERT deficiency caused significant reduction in pulmonary fibrosis along with diminished lung expression of pro‐fibrotic cytokines. Additionally, LysM‐TERT KO mice showed diminished numbers of c‐kit+ lung hematopoietic progenitor cells (LHPC) after BLM treatment in comparison with WT controls. Taken together these findings suggest a pro‐fibrotic role for the induction of TERT in myeloid cells.Support or Funding InformationThis study is supported by NIH grants HL052285 and HL112880.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Chronic Ethanol Intake on High‐Fat Diet Induced Metabolic Dysfunction in Mice
Authors: Coker Caitlin R. , Bingaman Sarah S. , Arnold Amy C. , Silberman Yuval 
Published: June 2021

High‐fat diet (HFD) induced obesity is associated with insulin resistance and is a major risk factor for numerous co‐morbidities such as type II diabetes. Conflicting clinical and preclinical evidence suggests that moderate alcohol consumption improves insulin sensitivity in obese subjects, and therefore may act to protect against obesity‐related metabolic comorbidities. However, the impact of free choice ethanol (EtOH) intake and HFD consumption on insulin action and glucose homeostasis in animal models remains unclear. Therefore, the overall goal of these studies was to determine if long‐term EtOH intake may alter insulin and glucose function in a mouse model of HFD induced obesity. Six‐week old male C57Bl/6J mice were given ad libitum access to HFD (n=10; 60% calories from fat) or control diet (Chow, n=10; 10% calories from fat) for two weeks. Mice were then given increasing volumes of EtOH (3%, 7%, and 10%) over one week as an EtOH‐ramp initiation period for subsequent 24‐hour access to a two‐bottle choice of water or 10% EtOH (vol/vol, in water). Body mass, EtOH and water intake, and EtOH preference was assessed every 24 hours for six weeks. HFD mice had significantly higher body mass compared to Chow mice (p<0.001). By the end of the study, Chow mice consumed significantly more EtOH than mice on HFD (p<0.001) and had higher preference towards EtOH (p<0.001). At the end of treatment, we measured 4‐hour fasting glucose levels and performed insulin tolerance tests (to examine the ability of intraperitoneal insulin to suppress blood glucose levels) in half of the mice in each group. These metabolic data were compared to weight and aged matched mice given HFD or Chow without EtOH access. Insulin sensitivity was significantly reduced in HFD mice (p=0.003; two‐way ANOVA), and EtOH consumption did not alter HFD‐induced insulin resistance. Furthermore, there appeared to be a trend for increased 4 hour fasting glucose levels in HFD (185.8±9.4), HFD+EtOH (176±25.9) and Chow+EtOH (177.8±21.6) groups compared to Chow alone (156.4±6.2). Together, these results suggest that moderate EtOH drinking in a two‐bottle choice model does not improve insulin sensitivity in HFD mice, but that higher levels of EtOH intake in control diet mice may cause impaired glucose homeostasis that may not be due to changes in insulin sensitivity.Support or Funding InformationPenn State Dept of Neural and Behavioral Science, NIH grants AA022937 and HL122507This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anticancer and pro‐apoptotic effects of Triphala extract in human breast cancer MCF‐7 cells
Authors: MURTHY MALLADI SATYANARAYANA 
Published: June 2021

Triphala, an ayurvedic formulation, showed a substantial antiproliferative effect against various human cancer cells including breast cancer cells. In vitro, anticancer and pro‐apoptotic studies were carried out using MCF‐7 breast cancer cell line to demonstrate its anticancer activity. Cell proliferation by MTT on Human breast cancer cells (1×106/ml) Flow cytometric cell cycle analysis, ELISA based quantification of cytosolic cytochrome c, Bax and Bcl‐2 levels and colorimetric assays of caspases‐3 and ‐9 were performed using breast cancer cells to demonstrate the pro‐apoptotic activity of triphala.Triphala showed substantial anti‐proliferative activity. In pro‐apoptotic studies, sub‐G0 cell phase population elevation, an increase of cytosolic cytochrome c and Bax levels, decrease of Bcl‐2 levels and enhanced caspases‐3 and ‐9 activities were observed in triphala‐treated MCF‐7 breast cancer cells. The results concluded that anticancer and apoptosis‐inducing potential of triphala in MCF‐7 breast cancer cells. Further detailed studies are in progress.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
3D Imaging and Visualization of Zebrafish and Giant Danio Hearts by Serial Block Face Scanning Electron Microscopy
Authors: Coffing Gabrielle , Hester Kamil , Kyaw Pyi T. , Lafontant Pascal J. 
Published: June 2021

3D and 4D imaging of zebrafish heart can provide significant insights into the structural and dynamic nature of the development and growth of this important biomedical model. Transmission electron microcopy (TEM) and tomography allow high resolution ultrastructural imaging of subcellular structures, adding to our understanding of heart architecture in model organisms. Serial block face scanning electron microscopy (SBFSEM) is a novel approach to nanoscale volumetric imaging of tissues and cells. To test the usefulness of SBFSEM in investigations of structure‐function relationship in zebrafish and giant danio hearts, we acquired over 3000 serial block face scanning electron micrographs at 50 to 100 nm intervals, as well as transmission electron micrographs, of the hearts of these fish. We subsequently extracted anatomical and cellular structures of interest in larval and adult fish by semi‐automated segmentation, and generated 3D visualization of various heart regions. Our results show structural similarity in the integration of the compact and spongy hearts in these two species. Similar to the zebrafish, the adult giant danio possesses transitional cardiac myocytes and a network of cardiac fibroblasts in the junctional region. Ultrastructural analysis also reveals remodeling of the ventricular junctional space during repair and regeneration, as well as the presence of luminal lacunae and persistent intra‐myocardial space as late as 3 weeks post‐fertilization. Taken together, our studies demonstrate SBFSEM as a powerful approach to gather nanoscale volumetric structural data in developing, adult, and regenerating giant danio and zebrafish heartSupport or Funding InformationNIH R15 HD084262‐01This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Organ Regeneration and Anti‐Aging Strategies
Authors: Belmonte Juan Carlos Izpisua 
Published: June 2021

Aging can be defined as the progressive decline in the ability of a cell or organism to resist stress and disease. Recent advances in cellular reprog‐ ramming technologies have enabled detailed analyses of the aging process, often involving cell types derived from aged individuals, or patients with premature aging syndromes. In my talk I will discuss how cellular reprog‐ ramming allows the recapitulation of aging in a dish, describing novel experi‐mental approaches to investigate the aging process. Finally, I will explore the role of epigenetic dysregulation as a driver of aging, discussing how epige‐netic reprogramming may be harnessed to ameliorate aging hallmarks, both in vitro and in vivo. A better understanding of the reprogramming process may indeed assist the development of novel therapeutic strategies to extend a healthy lifespan.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Shark EvoDevOmics: Linking Teeth and Taste Buds to the Evolution of Tooth Regeneration
Authors: Fraser Gareth 
Published: June 2021

The shark dentition is continuously regenerative, with teeth forming from a continuous and permanent epithelial dental lamina. We have identified an intriguing link between the regenerative potential of the shark dentition and the oral taste bud field, both of which form from a common epithelia. We performed RNAseq analyses on specific cell compartments within the continuous dental lamina and cells from the taste‐tooth (lamina) junction, associated with progenitor cells linked to tooth regeneration. We have identified a set of candidate genes involved in the initiation and maintenance of continuous dental regeneration. Expression analyses have verified these candidates and their relationship with specific compartments of the dental lamina associated with tooth regeneration. Our data provide a framework from which we can further investigate stem/progenitor candidates crucial for the process of tooth regeneration, in addition to providing new information on the timing and cyclicity of the shark tooth system. The dental lamina in sharks is an ideal regenerative organ in which to investigate perpetual tooth development, and to further our understanding of the origin of teeth. With these data we can start to construct gene regulatory networks as a tool to further investigate the functional role of these novel candidates. Our findings shed light on the developmental regulation of both the timing of continued dental initiation and the maintenance of stem progenitors for rapid tooth regeneration in an exciting and emerging EvoDevOmic model, the shark.Support or Funding InformationThis research was supported by the following research grants: Natural Environment Research Council (NERC) Standard Grant NE/K014595/1 (to G.J.F); and Leverhulme Trust Research Grant RPG‐211 (to G.J.F.).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Adapting the Chemistry and/or Biology of Proteostasis to Ameliorate Protein Aggregation Diseases
Authors: Kelly Jeffery W. 
Published: June 2021

The cellular protein homeostasis, or proteostasis network, regulates proteome function by controlling ribosomal protein synthesis, chaperone and chaperonin mediated protein folding, protein trafficking, protein degradation and related processes. Stress responsive signaling pathways match proteostasis network capacity with demand in each subcellular compartment to maintain or alter cellular homeostasis. The beginning of the seminar will focus on how the proteostasis network can be adapted pharmacologically through unfolded protein response arm‐selective signaling to alleviate the gain‐of‐toxic‐function diseases, including light chain amyloidosis and the transthyretin amyloidosis, where excessive secretion of misfolding and aggregation of proteins leads to a degenerative phenotypes. This strategy will be contrasted with high affinity small molecule binding to the normally folded structural ensemble of an aggregation‐prone protein inside and/or outside of the cell to stabilize the native state, lowering the population of misfolded, misassembly competent states that lead to aggregates, including amyloid fibrils. Lastly, we will cover the progress made to discover autophagy activators. These drug candidates are envisioned to be generally useful for ameliorating multiple neurodegenerative diseases based on human genetic evidence.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cracking the Glycocode: Finding the Signal in the Noise
Authors: Mahal Lara K. 
Published: June 2021

Glycans both encode cellular information, such as cell‐cell interactions and cell state (Signal), and must avoid being targeted by pathogens (Noise). This leads to a system in which the sugar code (i.e. the glycan motifs controlling function) is hidden within the noisy milleu of larger heterogenous glycan structures. This talk focuses on use of high‐throughput analytical methods, including our lectin microarray technology and newly developed miRNA‐proxy approach, in tandem with data integration, to decode structure‐function relationships in the glycome. Our work is identifying glycan drivers of biological function including those involved in melanoma metastasis and host‐response to pathogens (e.g. HIV‐1 and influenza), providing a host of new targets for small molecule intervention in these disease states.Support or Funding InformationThis work is supported by: NIH (1U01CA221229‐01, U01 AI111598) and the Melanoma Research Foundation.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Obesity, Insulin Resistance and Cancer: The PI3K connection
Authors: Cantley Lewis C. 
Published: June 2021

Retrospective studies have revealed that cancer rates in certain organ sites increase with obesity and some studies have suggested that the subset of obese individuals who develop insulin resistance are more likely to develop cancers than those who are obese but remain insulin sensitive. The risk of developing uterine and breast cancers are particularly high in women who are obese and insulin resistant and these two cancers have the highest rates of activating mutations in PIK3CA, the gene that encodes phosphoinositide 3‐kinase 1 alpha (PI3K1alpha). PI3K1alpha mediates insulin dependent cell growth and the activating mutations in PIK3CA allow this enzyme to be more readily activated by insulin in tumors that express the insulin receptor. Normal endometrial and breast epithelial cells express the insulin receptor and tumors in these tissues often have elevated expression of the insulin receptor. These observations raise the possibility that the elevated serum insulin in patients who are insulin resistant may be contributing to the development of tumors, especially tumors in tissues with PI3K pathway mutations. Many PI3K inhibitors have entered clinical trials and some have been approved. However, since the inhibitors that target PI3K1alpha cause insulin resistance in liver, muscle and fat, they cause hyperglycemia and hyperinsulinemia. Our studies indicate that the elevated insulin in the serum can ultimately re‐activate PI3K1alpha in the tumor, compromising the effect of these drugs. Strategies for making PI3K inhibitors more effective by preventing this feedback reactivation of PI3K1alpha in tumors will be discussed.Support or Funding InformationSupported by the NCI, the Breast Cancer Research Foundation, the Gray Family Foundation and the Hertog family.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Metalloenzyme‐catalyzed carbon‐carbon bond formation in natural product biosynthesis
Authors: Seyedsayamdost Mohammad R. 
Published: June 2021

Microbial secondary metabolites continue to inspire chemists and biologists alike with their fascinating structures and potent biological activities. Investigations into their biosynthetic pathways not only provides possible means of creating these molecules in the laboratory but can also reveal new enzymatic reactions and their underlying mechanisms. In this talk, I will present new transformations that my group has discovered in the biosynthesis of complex natural products, with a focus on carbon‐carbon bond formation catalyzed by metalloenzymes. Our efforts are expanding the scope of reactions that microorganisms have at their disposal in concocting bioactive small molecules.Support or Funding InformationThis work was supported by the National Institutes of Health (DP2‐AI‐124786) and the Burroughs Wellcome Fund.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
IL‐36γ Promotes Alveolar Macrophage Survival During Influenza Infection, Limiting Morbidity and Mortality
Authors: Wein Alexander N. , Dunbar Paul , McMaster Sean R. , Denning Timothy L. , Kohlmeier Jacob E. 
Published: June 2021

Influenza viral illness has been a high priority research topic in public health and immune pathogenesis, but the roles of individual cytokines in controlling the infection remains unclear. The IL‐36 family of proteins is made up of novel IL‐1 family members that have shown key roles in lung immunity to bacterial infections, but the role of IL‐36γ in influenza infection is unknown. We investigated the function of IL‐36γ during infection with low and high pathogenesis stains of influenza Il36g mRNA is upregulated in the lung following influenza infection with both H1N1 and H3N2 influenza. Genetic deletion of Il36g results in greatly increased morbidity, mortality, and viral titers compared to mice replete for the gene. This difference correlates with increased pro‐inflammatory cytokines in the airways of mice early during infection. Flow cytometric analysis of the immune compartment revealed a dramatic loss of alveolar macrophages by three days post‐infection. Previous studies have shown that alveolar macrophages have a key role in limiting early viral replication and preventing death during lethal influenza infection. We observed that the alveolar macrophages in Il36g−/− mice have an M2‐like phenotype at baseline, possibly contributing to their rapid depletion. Transfer of WT alveolar macrophages to Il36g−/− mice prior to influenza infection leads to increased survival compared to mock‐transferred Il36g−/− mice. These data indicate that IL‐36γ is a key protein in driving alveolar macrophage survival during influenza infection and protecting against severe disease.Support or Funding InformationResearch Support: R01 HL122559This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.