bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of Dihydromotuporamine C Derivatives on Actin Assembly Dynamics
Authors: Heidings James Bernard , Mathin Amirah , Skruber Kristen , Muth Aaron , Phanstiel Otto , Kang Hyeran 
Published: June 2021

Dihydromotuporamine C (motu C) is a small molecule isolated from a New Guinea sea sponge that has demonstrated anti‐metastatic and anti‐migratory properties both in vivo and in vitro. Many cancers do not pose a threat to a patient's life until they metastasize. Actin filament assembly dynamics play a critical role in cancer cell metastasis, cancer cell movement, and force generation. In this study we investigate how motu C derivatives modulate actin filament assembly kinetics in vitro, using biophysical and biochemical assays. The effect of motu C on average filament lengths was determined by direct visualization of actin filaments incubated with motu C derivatives, using total internal reflection fluorescence (TIRF) microscopy. We demonstrate that one particular motu C derivative, motuCH233, reduces filament length significantly. Pyrene actin assembly kinetics indicate that motuCH233 also effectively decreases the amount of polymerized actin, which may potentially affect cancer cell migration. Outcomes from this project can be used to further optimize the molecular design of anti‐migratory small molecules.Support or Funding InformationThis study was supported by the UCF start‐up fund and In‐House grant for Hyeran Kang.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Structural and Dynamics of Glucocorticoid Receptor Ligand Binding Domain in Complex with PGC1α and Synthetic Glucocorticoids
Authors: Liu Xu , Wang Yashuo , Ortlund Eric 
Published: June 2021

Synthetic glucocorticoids are important pharmaceutical drugs that are known to bind to the ligand binding domain (LBD) of glucocorticoid receptor (GR), a member of the nuclear receptor superfamily. These binding will induce conformational changes within GR for the subsequent DNA binding and different co‐regulators recruitments, ultimately lead to activate or repress target gene expression. One of the crucial co‐regulators is peroxisome proliferator‐activated gamma coactivator 1‐α (PGC1α), known to regulates energy metabolism by direct interacting with GR to modulate gene expression. However, the mechanisms through which PGC1α changes GR conformation to drive transcription are unknown. Here, an ancestral variant of GR (GR2) was utilized as a tool to produce stable protein for biochemical and structural studies. We report the first high resolution X‐ray crystal structures of GR2 LBD in complexes with PGC1α and dexamethasone and hydrocortisone. These structural analyses reveal how distinct steroid drugs bind to GR with different affinities via unique hydrogen bonding and hydrophobic interactions. Hydrogen‐deuterium exchange mass spectrometry showed different effects on GR2 LBD allosteric communication after glucocorticoids binding. Molecular dynamics simulations and network analysis identified the key residues involved in the intramolecular communication between ligand binding site and activation function surface, which is the PGC1α binding site.Support or Funding InformationThis work was supported by W.M. Keck Foundation Medical Research Grant to E. A. O. X. L. was supported by an American Heart Association postdoctoral fellowship [17POST33660110].This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Branched Amphiphilic Peptide Capsules: Agents for Gene Delivery and Immunomodulation
Authors: Tomich John M. , Sukthankar Pinakin R. , Barros Sheila de M. , Withaker Susan K. , Wilkinson‐Nutsch Kayla M. , Avila L. Adriana 
Published: June 2021

Branched Amphiphilic Peptides Capsules (BAPCs™) are a novel class of nano‐carriers constituted from the spontaneous co‐assembly of two unique, engineered peptide sequences. We recently reported on a thermally induced variant of BAPC™ that demonstrates promise as a vehicle for gene delivery and transfection. These BAPCs™ display a uniform size of 20–30 nm; they are easy to synthesize, stable and produce minimal immunogenic and inflammatory responses, in contrast to those commonly observed with viral and cationic lipid based approaches for gene delivery. BAPCs™ act as cationic nucleation centers allowing nucleic to wrap around them in a nucleosome like fashion, generating peptide‐nucleic complexes with sizes ranging from 50 to 250 nm. In this study, we combine the pre‐constituted BAPCs™ with a plasmid DNA construct encoding the green fluorescent protein (eGFP) in order to generate DNA/peptide complexes. We utilize these complexes to successfully transfect mammalian cells in vitro, yielding higher transfections rates than the commercially available lipid based reagent Lipofectin®, with minimal cytotoxicity. We then test the transfection system for the in vivo delivery of a DNA vaccine, previously designed to activate immune responses capable of controlling tumors induced by type 16 human papilloma virus (HPV‐16). We successfully demonstrate the efficacy of the DNA/BAPCs™ complexes in the in vitro activation of murine dendritic cells, and the enhancement of the associated vaccine's anti‐tumor potency, without significant toxic effects in vaccinated mice. Together these results indicate that the interaction of double stranded DNA to the cationic BAPCs™ nanoparticles represents a promising new in vitro and in vivo non‐viral gene delivery system.Support or Funding InformationPartial support for this project was provided by the Terry Johnson Cancer Center at Kansas State University, The Center of Excellence for Emerging and Zoonotic Animal Diseases (CEEZAD) at Kansas State UniversityThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Induction of Neoplastic Transformation, AP‐1 Signaling, PDCD4 signaling, and DNA Damage by Metal Oxide Nanoparticles Involves ROS‐Mediated MAPK Pathways in JB6 Cells
Authors: Barber Tabatha L. , Aldinger Joni , Arnold Joshua , Ding Min 
Published: June 2021

Occupational exposures to metal oxide nanoparticles have been reported to be harmful to human health, with possible risk of cancer among workers from many sectors. Metal Oxide nanoparticles have not, to our knowledge, been extensively examined for potential carcinogenic or genotoxic effects. To investigate the mechanisms of TiO2, WC‐Co, and CuO‐induced pathogenesis, the effect of metal oxide fine and nanoparticles on AP‐1‐MAPKs and ROS generation were investigated. The results indicated that CuO, WC‐Co, andTiO2 nanoparticles caused a 2‐fold increase or more in AP‐1 activity in JB6 cells. The induction of AP‐1 activity in cultured cell lines was time and dose‐dependent. The signal transduction pathways for AP‐1 activation were also investigated. Western Blot analysis demonstrate that CuO, WC‐Co, and TiO2 nanoparticles stimulate phosphorylation of p38 MAPK and ERKs. CuO, WC‐Co, and TiO2 also generated ROS when incubated with the cells as measured by electron spin resonance (ESR). Nano‐sized CuO, WC‐Co, and TiO2 generated more ROS than the fine sized particles when incubated with the cells. COMET assay suggests that exposure of the cells to CuO resulted in DNA damage. Cells pre‐treated with ROS inhibitors sodium formate and Polyvinylpyridine‐N‐Oxide (PVPNO) before exposure to CuO and WC‐Co nanoparticles demonstrate a significant reduction in p‐38 and ERK activity, suggesting the H2O2 may have a role in the ROS mechanism. Soft agar transformation assays demonstrated that there is a significant increase in colony formation in JB6/AP‐1 cells treated with CuO, WC‐Co, and TiO2 nanoparticles as compared to the control, indicative of neoplastic transformation. Unraveling the complex mechanisms associated with these events may provide insights into the initiation and progression of metal oxide‐induced pathogenesis.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Utilization of a DNA Nanoswitch to Detect Parallel Poladenylic Duplex Formation
Authors: Brylow Karl , Pickard Martha Anne G. , Anecelle Matthew R. , Gleghorn Michael L. 
Published: June 2021

Polyadenylic molecules (poly A) are single stranded nucleic acids containing a series of repeating adenine bases. Polyriboadenylic (poly (rA)) strands form parallel duplexed helices in acidic conditions. Our group has generated and utilized a technique to determine if polyadenylic sequences can pair when each sequence differs in composition. We have adapted a DNA nanoswitch developed by others to detect duplex formation where we can control the composition of each strand that is being tested for interaction. This method is superior to traditional UV melting studies, given the low specificity for detection of duplex species in a complex mixture of polyadenylic strands with different compositions. Our results show that nanoswitches can be used to detect poly(rA) duplex formation as a function of pH and we have identified interesting binding between polyadenylic strands that have different compositions. Our methodology not only provides fundamental information regarding parallel polyadenylic duplex formation, but also can help in design of future and potentially pH‐sensing biological nanotechnology.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular Beacon‐based Fluorescence Magnetic Nanoprobes for Tumor‐related HSP90 mRNA In‐suit Detection and Imaging
Authors: Chen Zhongyuan , Feng Yi , Xie Xiaoxue , Shen Xue , Yang Hong , Wu Chuihui , Liu Yiyao 
Published: June 2021

Heat shock proteins (HSPs) are a group of molecular chaperones that help proteins folding to their native conformation, protect cells from stressful environments. However, certain HSPs are overexpressed by malignant cells to survive under various therapies, inhibited cancer cells from senescence, apoptosis, stabilized lysosome membrane and promoted autophagy. Therefore, real‐time detection of the expression level of HSPs in living cells is required for both clinical cancer diagnosis application and oncology research to have a better understanding in anti‐apoptosis mechanism of malignant cells. Recently, delivery of molecular beacons that detect tumor‐related genes into cancer cells via nanocarriers shows promising results in the field of nanomedical diagnostics. In this study, uniformly sized and shaped superparamagnetic iron oxide nanocubes (SPIONCs) were synthesized by thermal decomposition and transferred into the aqueous phase via surface exchange with DMSA. G4 PAMAM dendrimer and Pluronic‐P123 were successively linked on SPIONCs‐DMSA in order to increase positive charge meanwhile reduce cytotoxicity. High specificity and sensitivity molecular beacon that targeting HSP90α mRNA was designed and synthesized and was condensed on PAMAM‐functionalized SPIONCs to form nanoprobes. TEM, XRD, TGA, ξ‐potential, DLS, VSM, fluorescence spectroscopy, agarose gel electrophoresis were carried out to characterize the morphology, composition, and properties of the nanoprobes. The mean diameter of as‐synthesized SPIONCs is 11 nm and increased to 100 nm after forming nanoprobes, which is stable in both water and culture media. Proliferation and hemolysis assays of the nanoprobes show low cytotoxicity and good hemocompatibility. LSCM confirmed that after 24h incubation with cells, the nanoprobes were uptake efficiently and located inside lysosome, and successfully escaped to the cytoplasm after 48h incubation. More importantly, the fluorescence of nanoprobes was observed significantly only in the HSP90 overexpressed MDA‐MB‐231 cells, instead of HSP90 low‐expressed MCF‐10A cells, which literally “highlights” a difference between normal and malignant cells, therefore provides a promising nanoplatform for real‐time gene expression level analysis and imaging in living cells.Support or Funding InformationWe would like to thank the financial supports, in whole or in part, by the National Natural Science Foundation of China (31470959, 81671821, 81471785), the Basic Research Program of Sichuan Science and Technology (2017JY0019, 2017JY0217), and the Fundamental Research Funds for the Central Universities (ZYGX2016Z001) for financial supports.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Phosphorylation of PEA‐15 in HIV‐TAT Mediated Disruption of Blood‐Brain Barrier
Authors: Cabezas Andres , Hassan Sherouk , Crespo‐Flores Sergio , Wei Yufeng 
Published: June 2021

Trans‐activator of transcription, TAT, one of the proteins that is expressed by the HIV genome, has been heavily associated with HIV‐associated cognitive morbidities. In vitro experiments have shown that the presence of TAT disrupts blood‐brain barrier (BBB) tight junctions and overall integrity. The mechanisms for TAT associated BBB disruption remain largely unknown.Phosphoprotein enriched in astrocytes (PEA‐15) is a ubiquitously expressed protein that is best known for its ability to regulate critical cellular pathways via protein‐protein interactions. PEA‐15 gains multiple protein binding affinities depending on the phosphorylation states of its residues Ser104 and Ser116. When PEA‐15 is phosphorylated at both Ser104 and Ser116 it gains high affinity to the Fas‐associated death domain (FADD). When protein‐protein binding with FADD, PEA‐15 blocks FADD from inducing apoptosis by preventing it from activating Caspase‐8.By blocking FADD effector functions via transfecting a phosphomimetic mutant of PEA‐15, in which both Ser104 and Ser116 are replaced with Aspartic Acid (PEA‐15‐DD), into primary human brain microvascular endothelial cells, we set to explore the mechanisms of HIV‐TAT mediated disruption of BBB tight junctions.Support or Funding InformationThis research is supported by NIH Grant R21DA046223This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Examining lateral development through CXCL14 modulation of CXCL12‐CXCR4 mediated gene expression in Danio rerio
Authors: Calderon‐Zavala Ariana Carolina , Wall Nancy 
Published: June 2021

The lateral line is a mechanosensory system used by fish to sense the movement of water. It is evolutionarily related to the inner‐ear in humans. For both organisms, the binding of the CXCL12 (SDF‐1 ligand) to the CXCR4 receptor induces conformational changes needed to activate signal transduction. This signaling results in numerous cellular responses such as cell fate, chemotaxis, and gene transcription. Interestingly, researchers have found that another signaling molecule, CXCL14, can also bind to the CXCR4 receptor with high affinity (Tanegashima et al., 2013). As a result, CXCL14 modulates CXCL12‐mediated chemotaxis, presumably acting as an allosteric regulator. In our lab, we our concerned with the allosteric relationship between CXCL14 and CXCL12 and how those relationships affect gene expression in lateral line development. To examine the effects of CXCL14 on CXCL12‐mediated gene transcription, zebrafish embryos were microinjected with CXCL14 antisense morpholino and incubated for a period of three timepoints: 24 dpf, 36 dpf, and 48 dpf, when lateral line development occurs. Our preliminary results suggest that the absence of CXCL14 affects the gene expression of CXCL14, CXCL12, CXCR4, CXCR7, epcam, claudin, and snail1b during lateral line development, thereby suggesting the allosteric capabilities of CXCL14. Further studies will be conducted to determine the degree to which CXCL14 affects the transcription levels of these genes.Support or Funding InformationWiscAMP‐NSF #1400815This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
An Evaluation of Palladin Overexpression in Pancreatic Cancer Cells
Authors: Bush Jason , Vander Karamjot , Chu‐Bacher Rowena , Ramirez Joel 
Published: June 2021

Pancreatic cancer (PC) is the fourth leading cause of cancer in United States with overall survival rates of less than 5% (ACS, 2018). Its aggressive nature and late onset of physical symptoms lead to poor prognoses. Recognition of biomarkers is clearly an imperative undertaking for the disease. To tackle this problem, we evaluated the secreted protein profile of pancreatic cancer cell lines derived from different stages. In total, nine human pancreatic cancer cell lines were successfully adapted into serum‐free environments from which conditioned media was collected by a combination of ultracentrifugation, molecular weight cutoff, protein precipitation, gel‐based separation, and tryptic digestion followed by mass spectrometry protein identification. A small subset of proteins has been identified and validated biochemically including Palladin (PALLD), a pancreatic cancer susceptibility protein shown to be up‐regulated in previous studies. PALLD is a scaffolding protein that supports remodeling of the actin cytoskeleton in association with integrins at focal adhesion sites to coordinate cell migration. We demonstrate a possible interaction between PALLD and the beta5 integrin (ITGB5) indicative of the increased motile phenotype in PC. Taken together, our results suggest that PALLD overexpression is associated with enhanced motility in pancreatic cancer cells and could represent a potential biomarker or therapeutic strategy against pancreatic cancer progression.Support or Funding InformationThis work was funded in part through intramural programs at Fresno State to KV, RCB, JAB, and through NCI P20CA138025 to JAB.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Neuronal membrane proteasomes target the nascentome to generate extracellular peptides that modulate neural network activity
Authors: Margolis Seth Shatkin , Ramachandran Kapil , Turker Fulya 
Published: June 2021

In mammals, activity‐dependent changes in neuronal function require coordinated regulation of the protein synthesis and protein degradation machinery. However, the biochemical evidence for this balance and coordination is largely lacking. To investigate this we initially used acute metabolic radiolabeling of stimulated primary mouse neurons to follow the fate of polypeptides being newly synthesized. We observed polypeptides being newly translated exclusively during neuronal stimulation were rapidly degraded by the neuronal membrane proteasome (NMP) and not the cytosolic proteasome. This turnover correlated with enhanced production of NMP‐derived peptides into the extracellular space which have the capacity to mediate neuronal signaling in part through NMDA receptors. Using in‐depth, global, and unbiased mass spectrometry, we identified the nascent protein substrates of the NMP. Among these substrates, we found that immediate‐early gene products c‐Fos and Npas4 were targeted to the NMP during ongoing activity‐dependent protein synthesis. Moreover, we found that turnover of nascent polypeptides and not full‐length proteins through the NMP occurred independent of canonical ubiquitylation pathways. We propose that these findings generally define a neuronal activity‐induced protein homeostasis program of coordinated protein synthesis and degradation through the NMP. This generates a new modality of neuronal signaling in the form of extracellular peptides with potential significance for our understanding of nervous system development and function.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Correlation Between Accumulation of Alzheimer's Disease Markers in Brain and Retina of Individuals with Late Stage of the Disease
Authors: Salloum Liza , Bondarenko Vladimir 
Published: June 2021

This study was aimed to determine if there is a correlation between accumulation of various Alzheimer's disease (AD) markers in the brain and retina of individuals in the late stages of the disease. Characterizing the distribution and accumulation of these markers in the retina and brain may be important for early diagnosis of neurodegenerative diseases such as AD.We examined total tau and phosphorylated tau protein accumulations in different areas of the brain and in retinal tissues from aged individuals with AD in order to compare the intensity and sensitivity of these biomarkers.Tissues from the hippocampus and temporal lobe of four different cadavers were analyzed. Two cadavers had diagnosed Alzheimer's disease, and two cadavers served as a non‐AD control. We utilized cryosectioning to prepare tissue sections for immunostaining. Tissues obtained from individuals without a history of Alzheimer's disease or other types of dementia were used as negative controls. Detection of tau was performed using a primary antibody against human tau proteins and amyloid‐b, followed by a universal secondary antibody.Although the staining for AD markers was greater than controls for both brain tissues, the density was higher in the hippocampus than in the temporal lobe. The density of the total tau (TT) was greater than that of phosphorylated tau (PT181) in both the hippocampus and temporal lobe. We also found this to be the case in the brain tissues of a control, indicating that total tau may be a more sensitive biomarker than phosphorylated tau.These data demonstrate that total tau may be a stronger biomarker than phosphorylated tau in quantifying protein accumulations in the brains of Alzheimer's disease cadavers. This raises the intriguing possibility that there may be more sensitive biomarkers that can better predict the course of neurodegenerative diseases such as AD. Thus, future studies will concentrate on the accumulation of tau in the brain and retina of individuals during the earlier stages of Alzheimer's disease, as well as the sensitivity and accuracy of different biomarkers.Support or Funding InformationTouro University Nevada intramural grant.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.