bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ischemia‐Reperfusion Induces Transcriptional Activation of Brain Natriuretic Peptide Gene via ERK1/2 in H9c2 cells.
Authors: Choe Yun‐jung , Ko Kyoung‐Won , Lee Sun‐Young , Youn Ho‐Joong , Kim Ho‐Shik 
Published: June 2021

Brain natriuretic peptide (BNP) is a cardiac hormone released from heart ventricular cells in response to myocyte stretch. Although BNP has recently been shown to be cardioprotective during myocardial ischemia/reperfusion (IR), mechanism of BNP gene expression by IR has not been clarified yet. Here, we show the mechanism underlying IR‐induced BNP gene expression in H9c2 rat cardiomyocytes.IR induced expression of BNP mRNA and transactivation of BNP promoter in H9c2 cells. Activation of MAP kinase family members such as ERK1/2 and p38 MAPK was triggered by IR. U0126, an inhibitor of MEK, but not SB203580, an inhibitor of p38 MAPK, completely prevented increase of IR‐induced BNP gene expression. In addition, antioxidants such as DMSO and TEMPO diminished induction of BNP and phosphorylation of ERK1/2. To investigate whether increase in free intracellular calcium [Ca2+]i has a role in expression of BNP by IR, calcium chelator BAPTA‐AM was treated before reperfusion. BAPTA‐AM strongly reduced induction of BNP by IR. Moreover, calcium ionophore A23187 increased levels of BNP mRNA and phosphorylation of ERK1/2. Taken together, IR‐mediated BNP gene expression may be due to calcium‐ and ROS‐mediated activation of ERK1/2 in H9c2 cells. This research was supported by MRC from cell death disease research center funded by KOSEF.
MAP kinase‐mediated regulation of DNA replication origin licensing and Cdt1 stability
Authors: Cook Jeanette Gowen , Chandrasekaran Srikripa , Tan Ting Xu 
Published: June 2021

Eukaryotic cells employ many mechanisms to ensure that their genomes are completely duplicated precisely once each cell cycle. One of the most important of these mechanisms is the regulation of origin licensing by the chromatin loading of the MCM complex through the concerted action of ORC, Cdc6 and Cdt1. Origin licensing control must be maintained even when cells are challenged with extracellular perturbations such as DNA damage or agents that trigger a cellular stress response.We have investigated effects on the origin licensing system mediated by the cellular stress response which is induced by a variety of insults such as hyperosmotic shock, exposure to bacterial toxin, or inflammatory cytokines. Unlike the effects of DNA damage, both Cdt1 and Cdc6 are stable during a cellular stress response; in fact Cdt1 is hyperstable and is no longer a substrate for DNA damage‐induced degradation. Instead, Cdt1 is robustly phosphorylated by the stress‐activated p38 and JNK members of the MAP kinase family. We further find that the activity of both p38 and JNK MAP kinases not only prevent de novo MCM chromatin loading during G1, but also induce rapid unloading of MCM complexes. The outcome of ongoing experiments with stably‐expressed mutationally altered forms of Cdt1 will shed light on these novel molecular consequences of stress kinase signaling.Supported by NIH GM083024 and CA094907.
Expression profiling of cellular transformation induced by tumor‐associated DNA polymerase beta variant I260M
Authors: Donigan Katherine Ann , Sweasy Joann 
Published: June 2021

DNA damage in vivo is estimated to occur upwards of 20,000 lesions per cell daily. Base excision repair (BER) resolves abasic sites and damaged bases. During BER, DNA polymerase β (Pol β) fills DNA gaps after excision of damaged bases. Approximately 30% of human tumors express Pol β variants not present in normal tissue. Many have altered fidelity and activity in vitro and induce cellular transformation. Prostate tumor variant I260M transforms cells and is a sequence‐context dependent mutator. To test the hypothesis that mutations induced in vivo by I260M lead to transformation, we characterized the genome‐wide expression profile of a clone expressing I260M compared to its non‐induced counterpart. Using a 1.5 fold cutoff with a false discovery rate of <0.05, 912 genes were found to be altered. Gene Ontology clusters were analyzed using Ingenuity Pathways Analysis to identify altered gene networks and associated nodes. Three nodes of interest with unaltered expression exhibited dysregulation of downstream targets. One node, peroxisome proliferator activator protein γ (PPARG), was found to have a coding mutation present only in transformed cells. PPARG is a transcription factor implicated as a tumor suppressor. This mutation may have contributed to driving cellular transformation. We conclude that I260M induces cellular transformation by a mutational mechanism.This work was supported by 5R01CA080830 to J.S.
Identifying IRES RNA‐Protein Complexes by SILAC based Mass Spectrometry
Authors: Tsai Becky Pin , Wang Xiaorong , Huang Lan , Waterman Marian L 
Published: June 2021

The dynamic nature of RNA protein complexes has made it challenging to study post‐transcriptional regulation in development and disease. We have developed a strategy to rapidly isolate and characterize in vivo assembled protein complexes for specific RNAs. Our proof‐of‐principle RNA target is a 1.2 kb IRES (internal ribosome entry site) in the 5′UTR region of Lymphoid Enhancer Factor‐1 (LEF1) mRNA. IRESes provide an alternative mechanism of ribosome recruitment and translation that is not well understood. To isolate in vivo assembled IRES‐protein complexes, the LEF1 IRES is engineered with a cluster of stem‐loops recognized by in vivo biotinylated MS2 bacteriophage coat proteins. The MS2 protein is used to rapidly isolate the complex with magnetic streptavidin beads.Our strategy is designed to isolate in vivo assembled RNA‐protein complexes for LC MS/MS identification of IRES binding proteins. SILAC‐based quantitative MS is used to detect proteins enriched on the target LEF1 IRES vs. non‐IRES mRNA. Results include known and putative IRES binding factors that were moderately enriched on the LEF1 IRES. The finding suggests that IRES elements assemble complexes of canonical RNA regulatory factors, rather than a few dedicated factors. In summary, our method provides a versatile and highly adaptable approach that can be applied to different RNA‐protein interactions.
Biophysical characterization of features of RNA helicase A that confer translational control of retroviral and selected cellular mRNAs
Authors: Ranji Arnaz K , Kvaratskhelia Mamuka , Boris‐Lawrie Kathleen A 
Published: June 2021

The 5′ untranslated region of some retroviruses and cellular mRNAs contain a cis‐acting Post‐transcriptional Control Element (PCE) that is necessary for efficient translation. Specific interaction of PCE with host RNA helicase A (RHA) is necessary for translation of the PCE‐containing viral and cellular mRNAs. This study sought to understand features of RHA necessary for selective interaction with the RNA switch. Site‐directed mutagenesis determined that the central DEIH helicase core is necessary for PCE activity, while the RGG domain is necessary for the nuclear localization. Here biophysical analyses tested the hypothesis that two redundant N‐terminal RNA binding domains are necessary to confer specific interaction with the Spleen Necrosis Virus (SNV) and junD PCE target RNA. Protein footprinting was employed to detect amino acids of RHA that interact with PCE RNA. Electrophoretic mobility shift assays and fluorescence polarization assays determined that the N‐terminal double‐stranded RNA binding domains specifically recognize structural features of PCE RNA. Loss‐of‐function point mutations in the double‐stranded RNA binding domains abolished binding to target mRNAs. The DEIH core does not exhibit affinity for the target mRNA, while the C‐terminal RGG domain binds non‐specifically to PCE RNA. Our results posit the model that PCE interaction with the N‐terminal domain of RHA exposes the DEIH domain, which catalyzes RNP rearrangement that facilitates polysome association. Molecular dissection of this new translational control axis may provide strategic drug targets for infectious disease and cancer.Funding: Glenn Barber fellowship of the OSU College of Veterinary Medicine and NIH R01 CA108882 and P01CA00730
Fatty Acid Suppression of Cell Proliferation is Associated with the Inhibition of Polyisoprenylated Methylated Protein Methyl Esterase
Authors: Amissah Felix , Taylor Shalina , Duverna Randolph , Lamango Nazarius S 
Published: June 2021

Protein polyisoprenylation is essential for the activities of oncogenic proteins such as Ras. The final, reversible step of polyisoprenylation pathway is catalyzed by an ester forming polyisoprenylated protein methyl transferase and polyisoprenylated methylated protein methyl esterase (PMPMEase). Various free fatty acids (FFAs) were analyzed to test whether their beneficial health effects are associated with PMPMEase inhibition. The potent inhibitory FFAs were further analyzed for their effects on cell viability. Arachidonic acid (AA) and lauric acid (LA) inhibited PMPMEase with 0.19 and 6.5 ìM, respectively. AA was over 63‐fold more inhibitory than Prostaglandin A2 (Ki of 12 ìM) while prostaglandin E2 and long chain saturated fatty acids were not inhibitory even at 1 mM. AA and LA inhibited the viability of human neuroblastoma cells with EC50 values of 11 and 25 ìM respectively, after 72 h exposure. Therefore AA and other PUFAs may be involved in the regulation of cell proliferation and that the elevated levels of COX‐2 in tumors convert PUFAs which inhibit PMPMEase at physiological concentrations, to prostanoids that are ineffective at inhibiting both PMPMEase and thus cell proliferation. NSAIDs may thus owe their anticancer effects through their ability to protect the PMPMEase‐ and cell proliferation‐inhibiting PUFAs from COX‐2 metabolism.Grant Funding Source: NIH/NIGMS/SCORE (GM 08111‐35) & NIH/NCRR (G12 RR0 3020)
Cyclin D1 regulates hepatic lipid metabolism
Authors: Hanse Eric Allan , Mullany Lisa K , Mashek Mara , Mashek Douglas G , Albrecht Jeffrey H 
Published: June 2021

Cyclin D1 regulates progression through the G1‐S phase transition of the cell cycle and plays an important role in liver regeneration. Cyclin D1 may also effect other cellular functions including metabolism. Here, we examined the role of cyclin D1 in hepatic liver fatty acid metabolism.METHODSMice were transfected with adenoviral vectors encoding cyclin D1 and cyclin D1‐KE, a mutant which does not induce cell cycle progression. In addition, primary rat hepatocytes and the differentiated mouse hepatocyte cell line AML12 were transfected with the same adenoviral vectors or siRNA targeting cyclin D1.RESULTSCyclin D1 regulated the expression of many mRNAs associated with lipid metabolism including downregulation of genes involved in lipogenesis (Srebplc, Fasn, Spot14 and Pklr). In cultured primary hepatocytes, overexpression of cyclin D1 (but not D1‐KE) led to decreased de novo lipogenesis and fatty acid incorporation into triglycerides and phospholipids. Cyclin D1 also promoted fatty acid oxidation and increased the import of fatty acids into hepatocytes. Cyclin D1 also inhibited the activity of Srebplc, Ppara, and Chrebp reporter constructs, whereas siRNA to cyclin D1 increased their activity in vitro.CONCLUSIONThese studies suggest that cyclin D1 regulates lipid metabolism in the liver, potentially through downregulating Srebp1c, Ppara and Chrebp activities.
Polymorphisms in UEA‐1 Lectin Binding to Human Buccal Cells and Salivary Fluid
Authors: Vanderpuye Oluseyi Adewale 
Published: June 2021

Human saliva may bind various microorganisms, but little is known about the surface proteins of buccal cells which are major constituents of this fluid. To further characterize buccal cell and cell‐free salivary fluid proteins and their variations among individuals, their reactivity with Ulex europaeus –I lectin (UEA‐1) was studied.After biotin‐UEA‐1 staining of nitrocellulose transfers of Laemmli SDS gels of 48 salivary fluid samples from different individuals, 42 had staining of the stacking gel. A 150 kDA band was present in 20 samples, a 75‐100 kDA was found in 22 samples and 6 samples had UEA‐1 staining in the region 40–50 kDa. Six out of 9 buccal cell samples had clear UEA‐I binding in the stacking gel of nitrocellulose transfers and differed in staining of 130kDa to 200 kDa proteins.Five of eight salivary fluid and 3 of four buccal cell samples coated onto microtiter plates bound biotin‐UEA‐1. By fluorescence microscopy, five out of seven buccal cell preparations bound UEA‐1. This study thus found differences in reactivity with UEA‐1 among different individuals. This polymorphism may be involved in differential binding of microorganisms to oral components.
Scorpion venom peptide accelerating the proliferation of irradiated M‐NFS‐60 cells
Authors: Dong Weihua , Qiu Yifang , Kong Tianhan 
Published: June 2021

Persistent myelosuppression occurring in some patients after radio‐ and chemotherapy is an indication of injury to HSCs. There is no available means to stimulate the full‐lineage blood cell recovery. M‐NFS‐60 cells (purchased from ATCC) can reflect proliferating characteristics of HSCs and were used in this study to evaluate the hematopoietic stimulating effects of SVP(separated from scorpion venom in our Lab). AlamarBlue assay, FCM, immunofluorescence and Western blotting methods were used to evaluate proliferating state, IL‐3 receptor (IL‐3R) protein expression and phosphorylating STAT5 levels in non‐irradiated or irradiated M‐NFS‐60 cells after it treated by SVP. The cell proliferation was accelerated, and the expression levels of IL‐3R and phosphorylating STAT5 protein were raised by SVP. The effects were more obvious in irradiated cells. SVP possessed co‐effects with IL‐3. The S phase percentage of M‐NFS‐60 cells was increased but cell cycle was retarded at G2/M phase in irradiated‐cells by SVP. It is suggested that SVP possesses hematopoietic growth factor‐resembling effects.Supported by National Natural Science Foundation of China Grant 30670906
The effect of sulfasalazine on homocysteine metabolism in vivo
Authors: Wu Yi‐Ting , Chiang En‐Pei Isabel 
Published: June 2021

Sulfasalazine (SSZ) is a disease‐modifying antirheumatic drug (DMARD) commonly used in the treatment of rheumatoid arthritis that inhibits intestine folate absorption and multiple folate‐dependent enzymes. The objective of the present study was to systematically investigate the impacts of long‐term SSZ treatment on homocysteine metabolism in vivo. C57BL/6J mice received either sterile 0.5% cellulose i.p. (vehicle, control group) or SSZ (80 mg/kg/day) for 8 weeks. Folate, S‐adenosyl methionine (adoMet), S‐adenosylhomocysteine (adoHcy) and homocysteine were determined in blood and tissues. Whole blood adoMet levels were significantly elevated compared to subjects on other DMARDs (1807 ± 1346, vs. 330 ± 861, p < 0.001), suggesting that SSZ may increase adoMet synthesis or decrease adoMet utilization. Furthermore, whole blood adoMet levels significantly correlated with the weekly dosage of SSZ (r = 0.34, p = 0.002) in patients receiving SSZ (n = 63).SSZ did not alter adoMet, adoHcy, methionine adenosyltransferase (MAT) activity and methylated cytidine content of DNA (dMC%) in mice liver or kidney. SSZ treatment increased whole blood adoMet level by induction of the methionine adenosyltransferase activity in erythrocytes in mice. In the present study we demonstrated the novel finding of a direct inducing effect of SSZ treatment on adoMet synthesis and adoMet synthase activity in blood.Grant Funding Source: ASN
Does lycopene or tomato powder impact lipid metabolism in CMO‐I KO and CMO‐II KO mice?
Authors: Elsen Amy Christine , Ford Nikki A. , von Lintig Johannes , Erdman John W. 
Published: June 2021

Adult mice lacking the &#946;‐carotene central cleavage enzyme, carotenoid‐15,15′‐oxygenase (CMO‐I), have altered lipid metabolism resulting in fatty liver development [Hessel at JBC 16;282(46):33553‐61]. The objectives of this study were to examine whether diet had any additional effects on lipid metabolism in CMO‐I KO female mice compared with female carotenoid 9′10′‐monooxygenase II knock‐out (CMO‐II KO) and wild type (C57BL/6Jx129x1/SvJ) mice. We fed an AIN‐93G diets consisting of 100mg/kg diet lycopene, 10% tomato powder, or placebo beadlets to knock‐out and wild type mice for 30 days. We evaluated body weights and compared the organ weights as a percentage of total body weight for each genotype. Preliminary data shows that CMO‐I KO uteruses were less than half the size of wild type uteruses and 1.5 times smaller than CMO‐II KO uteruses. Additionally, the effects of lycopene and tomato powder, and/or genotype on hepatic lipids were analyzed. We measured the expression of adipogenic genes, PPAR‐&#947;, PPAR‐&#945; and Acox1, in liver and adipose tissues. Results and implications of research will be discussed. (Funded in part by NIH grant PHS‐1‐RO1 CA125384.)Grant Funding Source: NIH
Cassia cinnamon supplementation before an oral glucose tolerance test in overweight or obese young women
Authors: Gutierrez Jean Lorraine , Cooke Matthew , Lutz Rafer , Rodney Bowden , Kane Robert , Willoughby Darryn 
Published: June 2021

ObjectiveThe purpose of this study was to compare the effects Cassia cinnamon and cellulose placebo on oral glucose tolerance test (OGTT) blood glucose (BG) and serum insulin (SI) values and the Matsuda insulin sensitivity index (Mat‐ISI) in young, healthy, overweight or obese and sedentary women.MethodsOn three separate days, 10 women (22.7 ± 4 years, BMI of 35.39 ± 5.36) reported for a fasted, venous blood draw. Participants consumed 5g of cellulose placebo or 5g of Cassia cinnamon bark. Three hours after the initial blood draw, the participants completed an OGTT by consuming a 75g glucose solution within 2 minutes. Venous blood draws were taken 30, 60, 90, and 120 minutes after OGTT to determine changes in BG and SI.ResultsThe maximum BG concentration following OGTT was significantly lower in the Cassia group, as compared to the placebo group (P = 0.025). The overall glucose response, glucose and insulin total area under the curve, and Mat‐ISI were not different between the two groups (P > 0.05).ConclusionsA 5g dose of Cassia cinnamon may reduce maximum post‐OGTT BG and improve glucose tolerance but does not have a significant impact on post‐OGTT insulin response in overweight or obese and sedentary women.
Compound K produces cardiac protection by activating Akt phosphorylation
Authors: Tsutsumi Rie , Mawatari Kazuaki , Katayama Erika , Nakaya Yutaka , Tsutsumi Yasuo M. 
Published: June 2021

Ginseng root is one of the most popular herbs throughout the world and it has been used as a medicine to enhance stamina and capacity to cope with fatigue and physical stress. In this study, we focused the cardiac protective effects of Compound K (C‐K; 20‐O‐D‐glucopyranosyl‐20(S)‐protopanaxadiol), which is a protopanaxadiol metabolite derived from ginsenoside Rb1, Rb2, and Rc by the metabolic actions of intestinal bacteria.Mice were exposed to 30 min ischemia followed by 120 min of reperfusion (control). Some mice were assigned to C‐K, or PI3K inhibitor, wortmannin. C‐K preconditioned significantly reduced infarct size compared to control group (22.4 ± 2.5% [n = 7] vs. 43.2 ± 2.0% [n = 12] respectively, p<0.05) and wortmannin eliminated the protection produced by C‐K (43.3 ± 2.4% [n = 7]). Additionally, the hearts treated with C‐K had a 2‐fold increase in basal phosphorylation of Akt and pretreated with wortmannin completely abolished Akt phosphorylation in response to C‐K treatment.These results reveal that C‐K produces cardiac protection in mice in association with PI3K/Akt signaling.
Effects of a single high‐fat meal rich in cottonseed oil on lipid profile, insulin sensitivity, and multimeric forms of adiponectin of healthy females
Authors: Sodhani Shradha , Sundaresan Sinju , Makkieh Kay , Imrhan Victorine , Mills Nathaniel , Vijayagopal Parakat , Prasad Chandan 
Published: June 2021

This study investigates the effect of consuming a single high‐fat meal rich in cotton seed oil (CSO) on plasma lipid, glucose, insulin, and adiponectin levels. In a randomized cross‐over design, eleven healthy female subjects consumed 1 of 2 iso‐caloric meals consisting of carrot muffin/s and milkshake providing 1 g of fat/kg body weight. Blended palm oil (BPO) served as control and blood drawn at baseline, 3 and 6 hours was analyzed for biochemical assays. One‐way repeated ANOVA was performed followed by post‐hoc tests to determine differences between three time periods and identify groups that were significantly different. Subjects were divided into groups based on the sequence of meal ingested (CSO followed by BPO) to account for carry‐over effects. Consumption of CSO did not show any significant differences (p<0.05) in mean blood glucose levels compared to BPO controls. The serum insulin levels also did not reach significance. No significant differences were observed in plasma levels of total cholesterol, HDL‐c, LDL‐c and triglycerides in CSO compared to BPO. CSO increased the ratio of polymeric to monomeric form of adiponectin compared to BPO (2.05 in CSO vs. 1.34 in BPO); however this increase did not reach statistical significance. A single high‐fat meal rich in CSO altered ratio of multimeric forms of adiponectin with no effect on insulin and lipid profile in healthy females.Supported by Texas Department of Agriculture
Chocolate matrix factors modulate pharmacokinetic behavior of cocoa flavan‐3‐ols following oral consumption by Sprague‐Dawley rats
Authors: Neilson Andrew Peter , Sapper Teryn N. , Janle Elsa M , Mattes Richard D , Rudolph Ralf , Matusheski Nathan V , Ferruzzi Mario G 
Published: June 2021

Little data exists on modulation of phase‐II metabolism of catechin (C)/epicatechin (EC) from chocolate by carbohydrate and milk. The objective of this study was to assess the impact of the matrix on profiles of plasma C/EC metabolites following administration of commercially relevant chocolate. Sprague‐Dawley rats were fed 1.5 g confection (ref. dark, high sucrose, or milk chocolate) by gavage; plasma was collected over 8 h. HPLC‐MS indicated that major metabolites of EC were EC‐5‐O‐β‐glucuronide, 3′‐O‐Me‐EC‐5‐O‐β‐glucuronide, and 4′‐O‐Me‐EC‐5‐O‐β‐glucuronide; major metabolites of C were C‐5‐O‐β‐glucuronide and 3′‐O‐Me‐C‐5‐O‐β‐glucuronide. Plasma metabolite concentrations were generally highest for high sucrose trt and lowest for milk treatment, while ref. dark treatment generally resulted in intermediate levels over 8 h. AUC of 4′‐O‐Me‐EC‐5‐O‐β‐glucuronide was significantly higher for high sucrose trt (2325 nM*h) vs. milk trt (1300 nM*h). Additionally, CMAX values for EC‐5‐O‐â‐glucuronide, 4′‐O‐Me‐EC‐5‐O‐β‐glucuronide, and 3′‐O‐Me‐EC‐5‐O‐β‐glucuronide were significantly higher for high sucrose trt (4012, 518, and 2518 nM, respectively) vs. milk trt (2590, 240, and 1670 nM, respectively). Milk and sucrose appear to modulate the metabolism and pharmacokinetics, and to a lesser extent overall bioavailability, of catechins from chocolate confections.Grant Funding Source: Kraft Foods research, Development, and Quality