bluuebunny/crossref_metadata_2025_split huggingface dataset:
Vitamin E analogs as chemotherapeutic agents and potentiators of chemotherapy in PC‐3 human prostate cancer cells
Authors: Graham Andrew , Oblad Richard , Barnes Bradley , Harmon Molli , Kenealey Jason 
Published: June 2021

Vitamin E ethyl acetate (VEEA),ethyl butyrate (VEEB), and succinate (VES) are three analogs of vitamin E that have anticancer activity. VES and VEA have been shown to selectively induce apoptosis in cancer cells across a number of cell lines via mitochondrial destabilization. We have shown that VEEA is more effective at reducing cell viability than VEEB and VES in PC‐3 human prostate cancer cells at 24, 48, 72, and 96 hour time points. Using mixture design response surface methodology (MDRSM), we have identified that 90% VEEA and 10% VEEB is the ideal combination for reducing cell viability at a 48 hour time point. We have also found that a mixture of VEEA and VEEB increases the effectiveness of chemotherapeutic drugs. We conclude that Vitamin E analogs have the potential to be chemotherapeutic agents, and increase the effectiveness of chemotherapeutics in PC‐3 human prostate cancer cells.Support or Funding InformationBryant Adams FoundationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inverse agonist of ERRγ inhibits the cell growth through the regulation of cell cycle and apoptosis in anaplastic thyroid cancer cells
Authors: Hwang Ji Sun , Cho Sung Jin , Hwang Hayoung 
Published: June 2021

In the previous study, we investigated that inverse agonist of ERRγ (estrogen‐related receptor gamma), GSK5182, enhances the responsiveness of radioiodine therapy by the modulation of sodium iodide symporter, NIS function in anaplastic thyroid cancer cells via the regulation of ERRγ and the MAPK signaling pathway. However, there is no reports showing the direct molecular mechanisms supporting the anti‐ cancer effect of inverse agonist of ERRγ. By the screening of inverse agonist of ERRγ, NDDC‐1228 which has more potent activity compared to GSK5182 was identified. Next, the functional activity of NDDC‐1228 was evaluated in the anaplastic thyroid cancer cells. At first, we evaluated the role of NDDC‐1228 in the regulation of thyroid cancer cell proliferation. NDDC‐1228 specifically inhibited the proliferation of CAL62, anaplastic thyroid carcinoma cell line under the cancer environment mimic condition but did not affect the proliferation of other organ derived cancer cell lines. When we examine the effect on the cell cycle, NDDC‐1228 treatment effectively induced the GI‐arrest by enhancing the expression of p21 and reducing the expression of Cyclin D and CDK proteins. In addition, NDDC‐1228 suppressed the expression of survivin, XIAP1 which are the inhibitors of apoptosis as well as activated the pro‐apoptotic protein, Caspase‐3. These results suggest that inverse agonist of ERRγ has anti‐cancer effects on anaplastic thyroid cancer cells via the regulation of cell cycle and apoptosis pathways and NDDC‐1228 could be used as a lead compound for the thyroid cancer therapy.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dynamics and Bifunctional Substrate Activation by an Arginine Drive Catalysis in Plant Chalcone Isomerases
Authors: Burke Jason , Noel Joseph 
Published: June 2021

In organic synthesis, bifunctional chiral catalysts are an increasingly popular tool to position and stabilize a nucleophile and activate an electrophile for asymmetric addition reactions. It turns out that organic chemists were not first to employ this synthetic strategy, plants have been using an similar method for quite some time (~500 million years). By using techniques in structural biology, enzymology and NMR, we characterize a novel mechanism by which the enzyme chalcone isomerase (CHI) catalyzes oxa‐Michael cyclizations of 2′‐hydroxychalcones into flavanones; a key early step in the synthesis of flavonoid compounds in plants. The enzyme enhances these reaction rates by utilizing the guanidine of a conserved arginine in a bifunctional capacity to position the nucleophilic phenoxide and activate the electrophilic enone of the chalcone substrate. In addition, we reveal the structural and biochemical basis of an evolutionary neofunctionalization event, in which a divergent clade of CHIs unique to Legumes expanded it's substrate repertoire to include 6′‐deoxychalcone substrates. Together these results reveal the details of key evolutionary mechanistic innovations unique to this class of plant enzymes.Support or Funding InformationHHMIThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The effects of ethanol extract of peros Punica granatum L. peel on the oxidative damage induced by diabet in rats
Authors: ÖMÜR Ali Dogan , Yıldırım Betül Dogan Apaydın 
Published: June 2021

This study is aimed to evaluate the effects of ethanol extract of peros Punica granatum L. peel on the oxidative damage induced by diabet in rats. In present study used twenty‐eight male Sprague–Dawley rats were randomly divided into four equal groups containing of seven rats per group. Group 1 kept as a control group, Received only vehicle 0.5 %CMC (peros), Group 2; streptozotocin group: Streptozotocin 60 mg/kg/ip single dose, Group 3; Punica granatum L peel extract (PGE) Group: PGE 10 mg/kg/20days/po, Group 4; Streptozotocin+PGE group: Streptozotocin as 60 mg/kg/ip single dose+PGE 10 mg/kg/20days/po. At the end of these processes, the animals were sacrificed, testis tissues were excised by dissection and evaluated as biochemical. Glutathione, malondialdehyde, catalase, glutathione peroxidase and superoxide dismutase were measured in the plasma and testis tissues. Punica granatum L peel extract was shown that the activity of catalase, superoxidase, glutathione peroxidase and glutathione activities increased and decreased levels of malondialdehyde in diabetic rats. As a result, the results showed that oral administration of Punica granatum L peel extract reduced the levels of lipid peroxidation and improved the antioxidant activity in plasma and testis tissues in rats treated with Punica granatum L. peel extract as compared with the diabetic and control rats. These results suggested that the Punica granatum L. peel extract enhanced the antioxidant defence under hyperglycaemic conditions hence protected the testes cells.Support or Funding InformationAtaturk UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Quorum sensing‐disrupting molecules to control virulence of Vibrio vulnificus, a fulminating foodborne pathogen
Authors: Choi Sang Ho , Kim Byoung Sik , Choi Garam , Jang Song Yee , Kim Myung Hee 
Published: June 2021

Bacterial pathogenicity involves numerous virulence factors required for successful survival in host environments as well as invasion and destruction of host tissues. The pathogenic marine bacterium Vibrio vulnificus is a causative agent of food‐borne diseases such as gastroenteritis and life‐threatening septicemia. It has been reported that expression of V. vulnificus virulence factors is well‐coordinated and regulated by sophisticated mechanisms consisting of many global regulatory proteins. Among them, SmcR, a quorum‐sensing master regulator and homologue of V. harveyi LuxR, has been identified and found to play a critical role in the regulation of the V. vulnificus virulence factors. The crystal structure of SmcR reveals that SmcR belongs to the TetR family protein possessing putative ligand‐binding pockets. To identify the small molecules which bind to and inhibit SmcR, chemical libraries were screened and an SmcR inhibitor exhibiting the most effective SmcR‐inhibition activity was selected and named QStatin. The lack of reduction of intracellular levels of SmcR in QStatin‐treated V. vulnificus suggested direct binding of the chemical to SmcR. Isothermal titration calorimetric (ITC) and crystallization analyses confirmed the chemical binding of QStatin into the pockets of SmcR. Although no big conformational changes were observed in the crystal structure, binding of QStatin reduced the DNA binding affinity of SmcR, and thereby changed the expressions of SmcR regulon similar to those of SmcR mutants. Accordingly, QStatin significantly inhibits expression of V. vulnificus virulence genes, leading to the decreased mortality of a brine shrimp infected with V. vulnificus. Most interestingly, QStatin attenuates pathogenicity of V. vulnificus, but does not inhibit growth itself at all, unlike existing antibiotics. Consequently, the combined results indicate that QStatin could be a novel lead compound to control QS‐mediated pathogenesis of V. vulnificus with the lowest possibility of the appearance of resistant strains.Support or Funding InformationThis work was supported by the National Research Foundation of the Ministry of Science, ICT, and Future Planning (2015R1A2A1A13001654 to S.H.C.).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Advantages and Disadvantages of Circadian Rhythms: A Tale of Two Microbes
Authors: Rust Michael 
Published: June 2021

Circadian clocks generate reliable ~24‐h rhythms despite being based on stochastic biochemical reactions. The circadian clock in Synechococcus elongatus uses a post‐translational oscillator that cycles deterministically in a test tube. Because the volume of a single bacterial cell is much smaller than a macroscopic reaction, we asked how clocks in single cells function reliably. Here, we show that S. elongatus cells must express many thousands of copies of Kai proteins to effectively suppress timing errors. Stochastic modeling shows that this requirement stems from noise amplification in the post‐translational feedback loop that sustains oscillations. The much smaller cyanobacterium Prochlorococcus expresses only hundreds of Kai protein copies and has a simpler, hourglass‐like Kai system. These altered dynamics are likely due to the loss of the KaiA protein in the Prochlorococcus system, which acts as a key node for the negative feedback loop in the system. Using advanced sampling techniques and molecular dynamics simulations, we find that KaiA acts as a nucleotide exchange factor by destabilizing the hexameric structure of the C‐terminal domain of KaiC. In KaiA‐less versions of this system, nucleotide exchange is presumably constitutive, potentially allowing the KaiC hourglass to act by measuring the ATP/ADP ratio in the cell. We show that this hourglass strategy can outperform a free‐running clock if internal noise is significant. This conclusion has implications for clock evolution and synthetic oscillator design, and it suggests hourglass‐like behavior may be widespread in microbes.Support or Funding InformationThis work was supported by NIH training grant T32‐GM007281, NIH F30 fellowship F30‐GM117962 (to J.C.), NIH R01‐GM107369 and a Pew Biomedical Scholars award (to M.J.R.).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Elimination of Wnt secretion from hepatic stellate cells impairs hepatocyte proliferation after partial hepatectomy
Authors: Ko Sungjin , Kim Min‐wook , Preziosi Morgan E. , Monga Satdarshan Pal 
Published: June 2021

Wnt‐dependent β‐catenin activation contributes to hepatocyte proliferation following 2/3 partial hepatectomy (PH) through regulation of Cyclin‐D1 (Ccnd1) expression, as we have previously shown using β‐catenin or Wnt coreceptors low‐density lipoprotein receptor‐related protein 5/6 conditional knockouts (KO). However, given the redundancy between the 19 mammalian Wnt proteins and various cell sources for Wnt secretion, the precise mechanisms of Wnt signaling in liver regeneration (LR) are not completely understood. Recently, using endothelial cell (EC)‐specific deletion of Wntless (Wls), a cargo receptor required for Wnt secretion, we have reported that sinusoidal ECs secrete Wnts (Wnt2 and Wnt9b) to activate β‐catenin in hepatocytes and enhance Ccnd1 expression, which is indispensable for proliferation of hepatocytes during LR after PH. Alternatively, conditional deletion of Wls in hepatocytes or cholangiocytes did not impact LR, whereas Wls deletion in macrophages only marginally affected LR post‐PH. Here, to investigate the roles for hepatic stellate cell (HSC)‐secreted Wnts in LR, we have characterized HSC‐Wls‐KO, generated by interbreeding Wls‐floxed and Lecithin Retinol Acyl Transferase (LRAT)‐driven Cre (Lrat‐cre) transgenic mice. HSC‐Wls‐KO mice showed no effect on liver weight or β‐catenin‐dependent pericentral gene expression at baseline. However, hepatocyte‐specific Cyclin‐D1 expression was abolished in baseline KO liver, whereas Cyclin‐D1 expression in non‐parenchymal cells was intact in KO liver. After PH, HSC‐Wls‐KO showed diminished protein and mRNA expression of Ccnd1, Ccnb1 and Ccna2 at 24–40 hour (h) post‐PH, but displayed slight increases at 72h, which led to a lower number of BrdU+ hepatocytes at 24 and 40h but a rebound increase by 72h. Interestingly, among several Wnts, HSC‐Wls‐KO livers at 24h post‐PH showed decrease of Wnt3 and Wnt9b mRNA compared to controls, although expression became comparable at 40–72h post‐PH.ConclusionAt baseline, HSCs secrete Wnt ligands essential for Cyclin‐D1 expression in midzonal hepatocytes but not for β‐catenin‐dependent hepatic zonation. Following PH, HSCs‐derived Wnts also contribute to Cyclin‐D1‐dependent hepatocyte proliferation during LR.Support or Funding InformationThe work was supported by NIH grants to S.P.M. (DK62277, DK100287, CA204586). S.P.M. is an Endowed Chair for Experimental Pathology.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regulation of expression of SG2NA isoforms in human cancer cell lines
Authors: Jain Buddhi Prakash , Goswami Shyamal K 
Published: June 2021

The Striatin family members (Striatin, SG2NA and Zinedin) have characteristic protein‐protein interaction domains as a caveolin‐binding domain, a coiled‐coil motif and a calmodulin‐binding domain at its N‐terminus and WD40 domain at carboxyl end. SG2NA was first reported in Lung and Bladder cancer patient serum as a Nuclear Autoantigen and its expression was found to be up regulated during the S and G2 phase of cell cycle. Mouse SG2NA generates five isoforms (87, 82, 78, 52 and 35kDas) by alternative splicing, intron retention and RNA editing. SG2NA isoforms are differentially expressed in mouse tissues viz., brain, heart, lung, liver and skeletal muscle. Besides alternative splicing and polyadenylation, sg2na expression is also regulated by post‐translational stability as its dephosphorylated form is more stable than the phosphorylated form. Further its role in Cancer progression and survival was established by its knockdown study. In the present study two splice variants (78 kDa and 87 kDa) of human sg2na were characterized. Further the expression of SG2NA isoforms in different human cancer cell lines (HEK293T, A549, H1299, HepG2, Hep3B, DU145, HELA) were assessed at the protein and RNA level which shows dyanamic expression. The expression of SG2NA isoforms varies among different cancer cell lines. In case of human cancer cell lines the expression of larger isoform i.e. 87 kDa was prominent unlike mouse which suggests that this SG2NA isoform in human is not undergoing proteasomal degradation. The ambiguity between mRNA and protein level in different cancer cell lines indicate its regulation at the post‐transcriptional or post‐translation level. There was no effect of proteasome inhibition on the stability of SG2NA in all the cells tested. The post‐transcriptional regulation study of SG2NA in different cancer cell lines would be a useful to show its expression and role in cancer progression.Support or Funding InformationDBT‐BUILDER, Government of India, awarded to the Jawaharlal Nehru University.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protein Disulfide Isomerase Inhibits and Reverses Alpha‐Synuclein Fibrillization
Authors: Ragusa Joey , Evangelista Baggio , Serrano Albert , Qiao Xin , Kim Yoon‐Seong , Chen Bo , Teter Ken 
Published: June 2021

Protein disulfide isomerase (PDI) is a ubiquitously expressed member of the thioredoxin family with related but independent oxidoreductase and chaperone properties. As a chaperone, PDI plays a crucial role in preventing the aggregation of misfolded proteins. Notably, PDI is known to inhibit alpha‐synuclein (SYN) fibrillization, a pathological hallmark of Parkinson's disease, but the mechanism by which PDI blocks the formation of SYN fibrils is unknown. Here, we report for the first time that PDI not only prevents SYN aggregation but is able to reverse aggregation both in vitro and in cultured cells. Assays using Thioflavin‐T fluorescence and transmission electron microscopy revealed that PDI is able to break down nascent but not mature fibrils. Furthermore, PDI was able to inhibit and reverse cellular fibrillization in HEK293T cells as assessed by protein‐fragment complementation and non‐reducing SDS‐PAGE. These data suggest a novel disaggregase function for PDI that disrupts the SYN fibrils most commonly found in Parkinson's disease. Current treatments for PD are limited to palliative care, but strategies focused on targeted therapeutic intervention are becoming popular. Thus, our work suggests the disaggregase property of PDI may lend itself to therapeutic development for Parkinson's disease.Support or Funding InformationThis work was supported, in part, by a LIFE at UCF Richard Tucker Gerontology Applied Research Award and a UCF College of Medicine competitive research 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.
Incorporation of Unnatural Amino Acids via Proteomically Informed Selective Pressure Techniques
Authors: Hill Austin Reece , Brown Amy , Boles Jeffrey Oakley 
Published: June 2021

Determining the three‐dimensional structure of a protein using x‐ray crystallography can be enhanced by complete or partial incorporation of unnatural amino acids into a target protein. Most naturally occurring atoms do not have sufficient mass to diffract electrons, therefore, heavy atom derivatives of target proteins are required. The Incorporation of unnatural metalloid containing amino acids provides a clear solution to the problem. A proteomic analysis of telluromethionyl Escherichia coli was carried out using the methionine auxotroph E. coli DL41(DE3) (pCock) expression system. Both telluromethionine and methionine exposed cultures were analyzed with the objective of identifying differentially expressed proteins. The analysis was carried out using 2D separation with isoelectric focusing serving in the first dimension and SDS polyacrylamide gel electrophoresis in the second. In‐gel tryptic digestion followed by peptide mass fingerprinting using nano‐ESI/qTOF/MS allowed identification of differentially expressed proteins. A few of the differentially expressed proteins identified as down‐regulated in systems exposed to TeMet were UPase, ZnuA, phosphoserine aminotransferase, and fructose‐bisphosphate aldolase. Understanding the biological roles of differentially expressed proteins was used to improve methods of TeMet bioincorporation using the selective pressure technique.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
DART‐ID Increases Single‐Cell Proteome Coverage
Authors: Chen Albert Tian , Franks Alexander , Slavov Nikolai 
Published: June 2021

Analysis by liquid chromatography and tandem mass spectrometry (LC‐MS/MS) can identify and quantify thousands of proteins in microgram‐level samples, such as those comprised of thousands of cells. Identifying proteins by LC‐MS/MS proteomics, however, remains challenging for lowly abundant samples, such as the proteomes of single mammalian cells. To increase the identification rate of peptides in such small samples, we developed DART‐ID. This method implements a data‐driven, global retention time (RT) alignment process to infer peptide RTs across experiments. DART‐ID then incorporates the global RT‐estimates within a principled Bayesian framework to increase the confidence in correct peptide‐spectrum‐matches. Applying DART‐ID to hundreds of samples prepared by the Single Cell Proteomics by Mass Spectrometry (SCoPE‐MS) design increased the peptide and proteome coverage by 30 – 50% at 1% FDR. The newly identified peptides and proteins were further validated by demonstrating that their quantification is consistent with the quantification of peptides identified from high‐quality spectra. DART‐ID can be applied to various sets of experimental designs with similar sample complexities and chromatography conditions, and is freely available online.Support or Funding InformationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identifying Colocalization of the Rho GTPase TCL/RhoJ with Coronin Proteins at Vesicular Membranes
Authors: McRoss Jasmine , Anliker Clair , Grigoryants Angelina , Samuelson Chelsea , Tader Brooke R , Hamann Michael J 
Published: June 2021

The Rho‐family GTPase TCL/RhoJ has been shown to enhance pro‐angiogenic and pro‐metastatic cellular processes; however, less is known about the signaling pathways and protein binding partners regulating its activity. TCL is part of the Cdc42 subfamily of Rho GTPases and bears a short, unique N terminus that is important for GTP‐loading and localization to the plasma membrane. Additionally, we have evidence that TCL is held to vesicular membranes through a protein/protein interaction when it is specifically in the GDP‐loaded confirmation. As several coronin family proteins bind other Rho family GTPases when they are in a GDP‐loaded conformation, we are evaluating whether coronins also selectively and spatially interact with GDP‐loaded TCL. To determine if the coronins co‐localize with TCL at vesicular membranes, mCherry‐fused versions of several Homo sapiens coronin proteins will be transfected into HeLa cells along with YFP‐tagged versions of TCL bearing constitutively active and dominant negative mutations. These TCL mutations will mimic GTP and GDP bound forms of TCL and help identify nucleotide dependent interactions with coronin proteins. Given the importance of TCL in tumor‐associated angiogenesis, these results will provide valuable insights into additional binding partners of TCL and may help clarify its role in pro‐angiogenic processes.Support or Funding InformationThis work was supported by Bemidji State University Biology Department and the College of Business, Mathematics, and Science. Support was also provided through the Neilson Foundation, Bemidji, MN and Regenerative Medicine Minnesota (RMM‐2017‐EP‐04).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.