bluuebunny/crossref_metadata_2025_split huggingface dataset:
Increasing Surface Expression of Pancreatic β cell‐KATP Channels Attenuates Palmitic acid‐induced Lipotoxicity in vivo and in vitro
Authors: Chen Pei‐Chun , Ruan Jing‐Syuna , Kuo Yi‐Ying , Chao Ru‐Yi , Chen Yun‐Wen 
Published: June 2021

In pancreatic β‐cells, KATP channels serve as glucose sensors and effectively initiate glucose stimulated insulin secretion (GSIS). When glucose enters β‐cell via glucose transporter 2, glucose metabolism increases intracellular ratio of ATP to ADP and in turn closes KATP channels which are composed of 4 sulfonylurea receptor 1 (SUR1) and 4 inwardly rectifying potassium channel 6.2 (Kir6.2) subunits. Chronic PA‐injected mice displayed type II diabetic characteristics including hyperglycemia and hyperinsulinemia. The pancreatic sections of these mice upregulated expression of cleaved caspase 3 and calnexin and importantly reduced surface expression of SUR1. Then, we tested time and dose effects of PA on the cell viability using INS‐1 cells. We observed the significant decrease in the surface expression of SUR1 after 72 hours of 0.4 mM PA. In addition, this treatment induced pancreatic b cells apoptosis by increasing cleaved caspase 3 proteins. Our results showed GB increased surface expression of SUR1 and decreased pancreatic β cell death and recovered insulin secretion upon glucose stimulation. Co‐treatment with GB in PA‐treated cells reduced ER stress and recovered mitochondrial membrane potential. Importantly, GB also improved glucose tolerance, insulin resistance, decreased triglyceride concentration and increased insulin release in PA‐injected mice. In conclusion, correct trafficking of KATP channels improved pancreatic β‐cell function and increased insulin sensitivity.Support or Funding InformationWe thank Dr. Christopher Newgard for the INS‐1 cell clone 832/13. We also thank the staffs of the Advanced Light Microscopy Core at National Cheng Kung University Hospital and Wallace Academic Editing for their help with this manuscript. This research received financial support from the Ministry of Science and Technology (MOST103‐2320‐B‐006‐005‐MY2, MOST105‐2628‐B‐006‐006‐MY3, MOST106‐2320‐B‐006‐050) offered to Pei‐Chun Chen.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impact of several PD‐associated genes on the toxicity of a‐synuclein in a yeast model
Authors: Jones Paul Andrew , Ong Emily , Balaram Ariane , Biel Alexsandra , Tembo Maiwase , DebBurman Shubhik 
Published: June 2021

Parkinson's disease (PD) is characterized by a‐synuclein misfolding and the death of midbrain neurons. PD can be described as familial, or sporadic, both of which are influenced by a multitude of environmental and genetic factors. Familial PD is directly caused by a mutation in one of at least ten genes, including SNCA, DJ‐1, VPS35, and ATP13A2. SNCA, which encodes a‐synuclein, has six identified missense mutations (A30P, E46K, H50Q, G51D, A53E, and A53T) that each cause autosomal dominant PD. Sporadic PD is linked with several risk genes and loci, including VPS13, the Sac I domain of SYNJ1, and the Swa2 domain of DNAJC6. Using our previously established budding yeast model system for α‐synuclein, we first show that wild‐type (WT), E46K, A53T, H50Q, and A53E a‐synuclein are toxic to yeast and show varying degrees of membrane binding and aggregation, while A30P and G51D a‐synuclein are relatively non‐toxic and shows cytoplasmic diffuse localization. What is still not well understood is whether the other PD‐causing and risk genes mentioned above can influence toxicity and localization properties of WT a‐synuclein and these six familial PD mutants. To test the hypothesis that they do influence a‐synuclein, WT and familial mutant forms of a‐synuclein were studied in haploid yeast strains that were singly deleted for these six PD‐linked genes (all of which are linked to loss‐of‐function in PD). Results show that some gene deletions increase (Δhsp31) or decrease (Δatp13, Δvps35) a‐synuclein toxicity and alter its localization in a highly familial mutant specific way, while others more broadly increase a‐synuclein toxicity or aggregation (Δvps13, Δsac1), while still others no effect (Δswa2). Our findings suggest that WT and each familial mutant of a‐synuclein create cellular toxicity and alter localization in distinct ways and that each is likely regulated by different subsets of genes, opening doors for mutant‐specific mechanistic insight into the varying modes of a‐synuclein toxicity.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of immobilization on the specificity and stability of choline oxidase
Authors: Fischel Adam H. , Holton Kortnie , Schmitz Jonathan M. , Watkins Linette M. 
Published: June 2021

Choline oxidase is a versatile enzyme that has uses in both industrial and medicinal settings. Choline oxidase is able to act as a biosensor and detect the presence of choline, as well as being able to produce glycine betaine, which is used in various industrial settings. In order for choline oxidase to be feasible in industrial synthesis pathways, it must have a broad specificity while maintaining activity over a range of pH and temperature. To this end, the specificity for two different substrates, temperature stability, and pH stability of choline oxidase were all determined. Following this, choline oxidase was immobilized through two different methods, by attachment to CnBr‐activated Sepharose beads, and by binding to a Polyethersulfone (PES) membrane. When immobilized with Sepharose beads, the temperature and pH stability of choline oxidase increased, while reducing the enzymes specificity for choline as a substrate at the same time.Support or Funding InformationThis work was supported by the James Madison University Department of Chemistry and Biochemistry and the Huntsman CorporationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification of the Substitutes on α‐(N)‐Heterocyclic Thiosemicarbazones that Interact with Human Topoisomerase‐II‐α with Nuclear Magnetic Resonance
Authors: Hill Jessica , Conner Jennifer , Combs Nathan , Bowman Sarah , Carroll William , Lisic Edward , Jiang Xiaohua 
Published: June 2021

Topoisomerase IIα (TopoIIα) is an essential enzyme for cell proliferation. Cancer cells are rapidly dividing cells. Thus, TopoIIα is a primary target for many anticancer drugs. Thiosemicarbazones (TSCs) have been known to possess anticancer activity by inhibiting topoisomerase‐II. Recent studies of our lab found the structure‐activity relationship of TSCs and their inhibition on TopoIIα. Our results suggest that the metal ion of TSC metal complexes and the terminal nitrogen side chain play important roles in increasing TopoIIα‐mediated cleavage complexes. The direct interaction between TSCs and TopoIIα remains unknown. It is hypothesized by our lab that the metal ions and the terminal nitrogen side chains interact with TopoIIα directly. This hypothesis is being tested by studying the interaction of 2‐acetylthiazole (ATZ) TSC, 2‐acetyl‐4‐methylthiazole (AMT) TSC, and benzoylpyridine (BZP) TSC metal complexes with TopoIIα by saturation transfer difference nuclear magnetic resonance (STD‐NMR) and diffusion ordered spectroscopy nuclear magnetic resonance (DOSY‐NMR). Recent STD‐NMR experiments suggest that the aliphatic region of 2‐acetylthiazole (ATZ) TSC and benzoylpyridine (BZP) TSC interact with TopoIIα.Support or Funding InformationThis research is supported by the Faculty Research Grant (XJ) and the NSF MRI project (1531870).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of methodology for the analysis of novel compound RpM
Authors: Fraser Andrew , Reddy E Shyam P. , Sarkar Arindam Basu 
Published: June 2021

ObjectiveThis study was carried out to develop the analytical methodology and limited characterization of the novel compound codenamed RpM. The structure of the novel molecule is being withheld to protect the intellectual rights and patent issues involved.IntroductionThe molecule under investigation, codenamed RpM, is a novel molecule that is custom manufactured for research purpose only. It is believed to have anti‐cancer activity and thus is a subject of interest and investigation.MethodThe UV‐Visble absorption spectra of the compound is established. A reverse phase liquid chromatographic method is developed to separate RpM from its close structural analogs code named RmBr, RpM and RPh using a Xorbax Eclipse XDB‐C18 as stationary phase. The mobile phase is a combination of acetonitrile and water used in a continuously changing concentration gradient. Standard curve was prepared between concentrations of 0.5–10 mcg/ml with RmBr as internal standard; limit of detection (LOD) and quantitation (LOQ) was established. Statistical analysis of the data was carried out. The solubility of the compound in water, water containing 5% DMSO and water containing 5% DMSO and 5% Cremaphor was established by vortexing and ultra‐centrifugation. The stability profile of the compound in water and water containing 5% DMSO and 5% Cremaphor is established in glass container.ResultsWithin the specified range the standard curve yielded a slope of 0.2231 (±0.0039) and an intercept of 0.0014 (±0.0058) and good linearity (R2≥0.999). RpM solubilities are found to be 0.1, 0.27, >5 mcg/ml in water, 5% DMSO and 5% DMSO+5% Cremaphor respectively. The compound is found to be stable (≥90% of original content) for at least 7 and days 5% DMSO and 5% DMSO+5% Cremaphor respectively. The stability is water is not determined because of the limited solubility of the compound in water.ConclusionThe project is successful in establishing an analytical methodology for RpM that is repeatable, robust, accurate and precise in the range. The solubility and stability of the compound in common aqueous based solvents have been established. More work is required achieve a complete physico‐chemical characterization of compound.Support or Funding Information*This work has been financially supported by University of Findlay, College of Pharmacy funds.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Physico‐chemical Characterization of Novel Compound RmBr
Authors: Higley Courtney , Sarkar Arindam Basu , Reddy E Shyam P. 
Published: June 2021

ObjectiveThis study was carried out to develop the analytical methodology and limited characterization of the novel compound codenamed RmBr. The structure of the novel molecule is being withheld to protect the intellectual rights and patent issues involved.IntroductionThe molecule under investigation, codenamed RmBr, is a novel molecule that is custom manufactured for research purpose only. It is believed to have anti‐cancer activity and thus is a subject of interest and investigation.MethodThe UV‐Visble absorption spectra of the compound is established. A reverse phase liquid chromatographic method is developed to separate RmBr from its close structural analogs code named RmBr, RmBr and RPh using a Xorbax Eclipse XDB‐C18 as stationary phase. The mobile phase is a combination of acetonitrile and water used in a continuously changing concentration gradient. Standard curve was prepared between concentrations of 0.5–10 mcg/ml with RmBr as internal standard; limit of detection (LOD) and quantitation (LOQ) was established. Statistical analysis of the data was carried out. The solubility of the compound in water, water containing 5% DMSO and water containing 5% DMSO and 5% Cremaphor was established by vortexing and ultra‐centrifugation. The stability profile of the compound in water and water containing 5% DMSO and 5% Cremaphor (a commonly used solubilizing agent) is established in glass container.ResultsWithin the specified range the standard curve yielded a slope of 55.62 (±1.57) and an intercept of −0.05 (±1.28) and good linearity (R2≥0.999). RmBr solubilities are found to be 0.055, 0.16 nd more than 5 mcg/ml in water, 5% DMSO and 5% DMSO+5% Cremaphor respectively. The compound is found to be stable (≥90% of original content) for at least 7 and days in both 5% DMSO and 5% DMSO+5% Cremaphor. The stability is water is not determined because of the limited solubility of the compound.ConclusionThe project is successful in establishing an analytical methodology for RmBr that is repeatable, robust, accurate and precise in the range. The solubility and stability of the compound in common aqueous based solvents have been established. More work is required achieve a complete physico‐chemical characterization of compound.Support or Funding Information*This work has been financially supported by University of Findlay, College of Pharmacy fundsThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Delivery of Interference RNA Molecules to Ovarian Cancer Cells Using Gold‐Liposome Nanoparticle Conjugates
Authors: Quiñones‐Díaz Blanca I. , Grafals‐Ruiz Nilmary , Barletta‐Bonanno Gabriel L. , Vivas‐Mejía Pablo E. 
Published: June 2021

RNA interference is a therapeutic modality in which RNA molecules, such as small interference RNAs (siRNAs) and microRNAs (miRNAs) (inhibitors and mimics), are used to regulate the expression of target mRNAs. MiRNAs are multitargeting RNA molecules that are involved in the regulation of several targets and can therefore influence multiple pathways when deregulated. On the other hand, siRNAs repress a specific gene by sequence complementarity to the target mRNA. Although both miRNAs and siRNAs have an increasing therapeutic potential, systemic delivery of RNA‐based technologies is limited by the poor stability of RNA molecules in circulation. Hence, our main goal is to develop a carrier for the efficient and targeted delivery of RNA molecules to ovarian cancer cells. To achieve this, we propose a nanocarrier formulation of gold nanoparticle (AuNP)‐RNA conjugates encapsulated on folate receptor targeted liposomes that could achieve a double protection layer for the RNA while increasing delivery to ovarian cancer cells that overexpress the folate receptor on their surface. We prepared carriers for siRNAs and miRNA oligonucleotide mimics by conjugation of the RNA molecules to the surface of 15 nm diameter AuNPs via thiol linkages. We performed this conjugation in presence of polyethylene glycol (PEG‐2000) to avoid gold precipitation. Physical characterization of the carriers was conducted by dynamic light scattering (DLS) and quantification of conjugated RNA was assessed by fluorescence. The conjugation resulted in approximately 47 to 75 RNA molecules linked per nanoparticle and conjugates of 20 to 25 nm diameter with a slightly negative charge. In addition, encapsulation of the AuNP‐RNA conjugates into liposomes was attained with more than 70% encapsulation efficiency. These results indicate the feasibility of the development and production of our nanocarrier. As a proof of principle we are using this nanoparticle conjugate to target Integrin‐linked kinase (ILK). ILK is a pseudokinase that has been found to be aberrantly expressed in ovarian tumors. Further experiments of internalization, stability and toxicity demonstrate the efficacy of this nanocarrier as a delivery system for miRNA mimics and siRNAs to ovarian cancer cells. We conclude that siRNA‐based technologies are an alternative strategy to target ILK and other target proteins lacking specific chemical inhibitors.Support or Funding InformationRCMI: U54 MD007600This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Validation of In Vivo Protein Surface Accessibility Method
Authors: Ma Michelle , Bamberger Casimir , Martínez‐Bartolomé Salvador , Yates John R. 
Published: June 2021

Measuring changes in surface accessibility is a way to probe conformational changes of proteins. Traditionally, in vitro methods of resolving protein structures have relied on the isolation and purification of proteins, but this artificial protein environment may not reflect its functional native conformation. Therefore, this project seeks to develop a novel approach to identify and quantify changes in protein surface accessibility caused by protein misfolding or altered biomolecule interactions in vivo on a proteome‐wide scale. We want to find out if this approach, chemically labeling the ɛ‐amino group of lysine residues, will yield the same results in vitro as it did in vivo, thus validating the accuracy of results generated with this technique. This dimethylation method is expected to yield biologically relevant information as it allows for the screening of proteomes for structural perturbations as well as identification of novel structural information for proteins not yet crystallized.Using GAPDH as a control, we showed that the GAPDH tetramer stays intact upon dimethylation using native gel electrophoresis. Using data from crystal structures of GAPDH, the GAPDH tetramer contains one predicted inaccessible lysine residue and we were able to detect this inaccessible lysine residue with our in vitro dimethylation experiment. In addition, under time‐limited conditions, the dimethylation reaction does not go to completion and appears to preferentially occur at certain lysine sites. The extent of dimethylation does not scale with relative surface accessibility and rather requires a minimal accessibility to the ɛ‐amino group of lysine residues.The dimethylation labeling method is reproducible in vitro as well as in vivo. The GAPDH experiments show that the method chemically labels proteins that are still in their native conformation. Limiting conditions of the dimethylation reaction provides additional structural information of the protein, which allow us to validate predicted structural models and/or data. This technique can be utilized to screen the proteome for structural alterations, which is especially important in diseases or drug interactions.Support or Funding InformationNIH NCI Enabling Technologies Grant (IMAT) R33CA212973‐01 to John R. Yates IIIThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Antiproliferative Effects of Soy Derived Dipeptides in Human Breast Cancer Cell Lines
Authors: Das Salil K. , Mitra Piyali , Maebuchi Motohiro , Mukherjee Shyamali 
Published: June 2021

In the United States, breast cancer is the second leading cause of cancer‐related deaths in women. Diet is an important factor in the development of numerous diseases, including breast cancer. We reported earlier that the consumption of soy protein controlled not only the proliferation of dimethylbenz [a] anthracene (DMBA)‐induced breast cancer in rats but also its aggressiveness. Soy protein is hydrolyzed in the gut to small peptides which are easily absorbed and are involved in various cellular functions. Fuji Oil Company, Osaka, Japan, has commercially marketed a soy protein hydrolysate, termed Hinute‐AM, composed of primarily di‐ and tripeptides, to be used as a health food. This product was found to have anti‐inflammatory property. It is possible that the antiproliferative effects of soy protein on breast cancer in rats may be mediated by its hydrolysis in the small intestine to small peptides. Hence, we hypothesized that the beneficial effect of soy protein in breast cancer development may be mediated by its hydrolysis product as peptides. Hinute‐AM was solubilized in 0.1% FA and analyzed by LC‐MS/MS. The data analysis has revealed the presence of five peptides (VPY, EL, IQ, LQ and YG) in Hinute‐AM. Out of these, three synthetic peptides (YG, LQ and IQ) were tested for their effects on proliferation of three normal (12A, 12F and 184A1) and four breast cancer cells (HCC1806, MCF 7, BT‐549 and MDA‐MB‐231). All peptides had significant antiproliferative effects in all breast cancer cell lines. However, the major effect was with YG and in particular in HCC1806 cell line. We reported earlier that expression of TSPO (translocator protein) is higher in breast cancer cells than normal breast cells. PCNA is also regarded as good cell proliferation marker. In this experiment, we studied the effects of YG on TSPO and PCNA expression in normal and three breast cancer cells (MCF‐ 7, BT‐549 and HCC‐1806 by immunohistochemistry. Results indicated that YG down‐regulated the expression of both TSPO and PCNA in all cancer cells. Furthermore, the binding capacity of 3H Ro5‐4864 to the TSPO receptors was significantly reduced by both YG and LQ. The mechanism by which YG and LQ inhibited the proliferation of breast cancer cell lines is under investigation. (Supported by grants from Fuji Oil Company, Osaka, Japan and NIH 5U54MD007593)Support or Funding InformationSupported by grants from Fuji Oil Company, Osaka, Japan and NIH 5U54MD007593This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.