bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of glutathione‐S‐transferases in the metabolism of the anti‐cancer agent and aromatase inhibitor, exemestane
Authors: Teslenko Irina L. , Chen Gang , Xia Zuping , Lazarus Philip 
Published: June 2021

Breast cancer is the most commonly diagnosed cancer in women, with approximately 1 in 8 women in the U.S. developing breast cancer over their lifetime. The majority of breast cancers diagnosed each year are estrogen receptor positive (ER+). Drugs targeting the ER pathway are a major treatment modality for ER+ patients and are additionally prescribed for the prevention of breast cancer. Exemestane (EXE) is an inhibitor which blocks the enzyme aromatase in the final step of estrogen biosynthesis. Studies examining the long‐term use of EXE as a chemopreventative agent demonstrated a reduction in breast cancer incidence by more than 65%. However, response and toxicity varied widely among individuals, which may be attributed to genetic polymorphisms in enzymes that metabolize EXE. Data from our lab demonstrate that a cysteine conjugates of EXE and its active metabolite 17β‐dihydro‐EXE (17β‐DHE) make up 77% of the total metabolite in urine. Cysteine conjugates are formed from metabolism of glutathione (GSH) conjugates in a reaction catalyzed by the glutathione‐S‐transferases (GST) family of the enzymes. The goal of the present study was to identify hepatic GSTs active in the metabolism of EXE and 17β‐DHE and to determine kinetic parameters for the identified enzymes. In a screening of the Protein Atlas, a total of 12 cytosolic and 3 microsomal GSTs were found to be hepatically expressed. EXE‐GSH conjugation activity in human liver samples was screened by incubating GSH with EXE or 17β‐DHE (0.125–1.0 mM) in cytosol (HLC) or microsomes (HLM) prepared from human livers. GSH conjugates were formed for both cellular fractions, indicating that both cytosolic and microsomal GST enzymes are active against EXE and 17β‐DHE. GSH conjugates were detected by UPLC‐MS and quantified using synthesized EXE‐GS and 17β‐DHE‐GS standards. The purity and identity of both standards and internal standards were confirmed using NMR and UPLC‐MS and found to be > 95% pure. Individual recombinant human GSTs were then screened to determine which enzyme is primarily responsible for the metabolism of EXE. Commercially available cytosolic GSTs were purchased, and their activity was verified with a common GST substrate, 1‐chloro‐2.4‐dinitrobenzene. Results from the individual EXE‐GSH conjugation assays indicate the cytosolic enzymes GSTA1, GSTM3 and GSTP1 are active against EXE and 17β‐DHE; studies of microsomal GSTs are on‐going. These data suggest that several GSTs, both cytosolic and microsomal, may be important in the phase II conjugation of EXE and its major active metabolite, 17β‐DHE.Support or Funding InformationThis study is supported by National Institutes of Health (NIH) (Grant R01‐ES025460 and R01‐CA164366) and the Health Sciences and Services Authority of SpokaneThis 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 Tumor Conditioned Media from Prostate Cancer Cells on Fibroblast Cell Morphology, Viability and Gene Expression
Authors: Kaur Sukhneeraj Pal , Cummings Brian S. 
Published: June 2021

Cancer is a heterogeneous collection of neoplastic cells, surrounded by non‐malignant, stromal cells, and extracellular matrix (ECM), among other constituents. Cross‐talk between tumor and stromal cells forms a dynamic tumor microenvironment (TME) that mediates cell growth, proliferation, and metastasis. The TME can act to inhibit cancer growth and proliferation during the initial stages of carcinogenesis. However, cancer cells are able to evade these inhibitory signals, and subsequently reshape the TME, making it more amenable to their own growth. This gives rise to numerous cancer‐associated stromal cells, including cancer‐associated fibroblasts (CAFs). CAFs further modulate the TME by regulating the release of growth signals, cell death, ECM remodeling, angiogenesis, and epithelial‐mesenchymal transition. To study the ability of prostate cancer to alter the TME in vitro, we exposed human bone fibroblasts (HS‐5 cells) to tumor conditioned media isolated from a human neuroendocrine prostate cancer cell line (DU145) and assessed changes in cell morphology, viability (using MTT assay) and the expression of Group IIA secretory phospholipase A2 (PLA2G2A), as well as its regulatory proteins, glypican‐1 (GPC‐1) and Group IVA phospholipase A2 (PLA2G4A), using qPCR. Exposure of HS‐5 cells to TCM from DU145 cells altered cellular morphology to a more mesenchymal phenotype typically seen in active myofibroblasts. TCM also decreased MTT staining in a time‐dependent manner that was independent of the presence of serum in the TCM. Changes in cell morphology and viability correlated to increases in the expression of PLA2G2A, GPC‐1 and PLA2G4A. The effect of TCM was time‐ and serum‐dependent, and was differential for each gene studied. Alteration in cellular morphology and increases in PLA2G2A, GPC‐1 and PLA2G4A in HS‐5 cells correlated to increases in the expression of fibroblast activation protein‐α, which is a CAF marker. To elucidate the mechanisms mediating changes in HS‐5 cells exposed to TCM, we investigated the expression of GLi1, a transcription factor involved in the activation of the sonic hedgehog signaling pathway. We found that TCM increased GLi1 mRNA levels in HS‐5 cells, with greater increases being observed using TCM containing serum. Collectively, these data demonstrate that media from DU145 cells can induce morphological and genotypic changes in human fibroblasts similar to those observed in CAFs. As such, this model holds promise for identifying molecular mechanisms by which prostate cancer cells alter the TME and induce prostate cancer cell growth.Support or Funding InformationNational Institute of Biomedical Imaging and Bioengineering NIBIB (EB0160100)Department of Defense Prostate Cancer Research Program Idea Development Award (PC150431 GRANT11996600)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
FOXO1 Couples Reactive Oxygen Species in Diabetic Atherosclerosis
Authors: LIU Jingjin , Yan Dan , Xie Xiang , XIA Zhengyuan 
Published: June 2021

Diabetic atherosclerosis, distinct from other type of atherosclerosis, is progressing rapidly which is the leading cause of morbidity and mortality in the entire world, and no effective therapy has been established so far. Thus, an intensive search for new therapeutic targets in diabetic atherosclerosis is needed. A growing evidences are that post translational modifications of FOXO1 triggered by hyperglycemia‐induced reactive oxygen species (ROS) which is most likely primarily responsible for hyperglycemia, plays an important role in diabetic atherosclerosis progression.Here, we investigated whether the inhibition of FOXO1 alters diabetes‐associated atherosclerosis. Our study found that pharmacologic inhibition of FOXO1 markedly suppressed the vascular remodeling in DM though attenuating overproduction of ROS, increasing antioxidant function in conjunction with enhancing antioxidant activity, and reducing ROS induced inflammation and fibrosis in carotid artery. Moreover, in T1DM rats, FOXO1 translocate into the tunica intima of carotid artery after treatment of pharmacologic inhibition of FOXO1, which coupled with the activation of PI3K/Akt/mTOR pathway. In addition, inactivation of PI3K/Akt/mTOR pathway in diabetic carotid artery was restored by pharmacologic inhibition of FOXO1 and the anti‐ROS effect of pharmacologic inhibition of FOXO1 was partially inhibited by a PI3K inhibitor (LY294002) and also enhanced p‐Akt/Akt protein ratio. Therefore, we hypothesis that pharmacologic inhibition of FOXO1 attenuate hyperglycemia‐induced ROS generation associated with activation of PI3K/Akt/mTOR pathway signaling, which may be a promising target for treating diabetic atherosclerosis and subsequent cardiovascular diseases.Support or Funding InformationRGC General Research Fund: GRF 17124614 (09/2014‐08/2017), GRF 17123915M (09/2015‐08/2018)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Antithrombotic Effects of 12‐LOX Derived Metabolites of DPA, ω‐6
Authors: Chen Angela , Yeung Jennifer , Szatkowski Anet , Jackson Madeline , Watson Josephine , Freedman Cody , Das Aditi , Holman Ted , Holinstat Michael 
Published: June 2021

IntroductionPlatelet‐mediated thrombosis is the primary underlying mechanism leading to cardiovascular life‐threatening clinical events. Regulating excessive platelet reactivity is an essential aspect of antithrombotic therapy. A number of anti‐platelet drugs have been developed to target specific signaling pathways or endpoints involved in platelet activation. Despite the effectiveness of current anti‐platelet therapies, uncontrolled thrombosis or bleeding complications still persist, warranting the development of novel anti‐thrombotic strategies. Previously, we had presented that 12‐HETrE, an oxylipin generated by 12‐lipoxygenase (12‐LOX) oxidation of ω‐6 polyunsaturated fatty acid (PUFA), dihomo‐g‐linolenic acid (DGLA), could be an approach to modulate platelet reactivity through the prostacyclin receptor. Here, we demonstrate that the elongated form of DGLA, docosapentaenoic acid (DPA; 22:5n‐6) can also be endogenously oxidized by platelet 12‐LOX to generate two distinct oxylipins, 11‐oxy‐DPA and 14‐oxy‐DPA, to regulate platelet reactivity.ObjectiveTo determine the efficacy of and the mechanistic target of 12‐LOX derived metabolites of DPA 11‐ and 14‐oxy‐DPA, ω‐6 in the regulation of platelet function.ResultsBoth 11‐ and 14‐oxy‐DPA potently inhibited platelet aggregation in an aggregometer measuring optical density. Treatment of whole blood with 11‐ and 14‐oxy‐DPA, ω‐6 also significantly reduced platelet adhesion as well as thrombus growth on an ex vivo collagen‐coated perfusion chamber under arterial shear. In support of these findings, using flow cytometry, calcium mobilization was attenuated in platelets treated with 11‐ and 14‐oxy‐DPA, ω‐6. Interestingly, both 11‐ and 14‐oxy‐DPA inhibited platelet activation in a Gas independent manner. Instead, 11‐oxy‐DPA and 14‐oxy‐DPA exerted their anti‐platelet effects through a peroxisome proliferated‐activated receptor (PPAR), by which pharmacological antagonism of the PPARα isoform reversed the ability of the metabolites to inhibit platelet aggregation. In supplement of these ex vivo findings, ongoing experiments are being performed to measure laser‐induced thrombus formation (platelet and fibrin) in the cremaster vessel of wild‐type mice intravenously administered with either 11‐ and 14‐oxy‐DPA.ConclusionThis is the first study to demonstrate the novel metabolites of DPA, ω‐6 generated by 12‐LOX also have a role in regulating platelet activation through the PPAR pathway. Our findings suggest that 11‐ and 14‐oxy‐DPA, ω‐6 shows great promise as a potential anti‐platelet therapeutic for the treatment for cardiovascular‐related diseases.Support or Funding InformationThis work was funded in part through the National Institutes of Health grants R01 GM105671 (MH), R01 HL114405 (MH), R01 MD007880 (MH).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mitochondrial respiratory function in the vasculature with advancing age: Examining the link to vasodilatory dysfunction.
Authors: Park Soung Hun , Kwon Oh Sung , park Song‐Young , Andtbacka Robert H. I. , Hyngstrom John R. , Richardson Russell S. 
Published: June 2021

BackgroundUnlike cardiac and skeletal muscle, the role of mitochondria in the vasculature is still unclear. Therefore, this study sought to document mitochondrial respiratory function in the vasculature with advancing age and examine the link to age‐related vasodilatory dysfunction.MethodsSkeletal muscle feed arteries (SMFAs) were harvested from young (30±6yrs, n=4) and old (69±8yrs, n=20) subjects. Mitochondrial respiratory function was assessed in permeablized smooth muscle fibers and vasodilation in SMFAs was assessed in response to flow‐induced shear stress, acetylcholine (ACh), and sodium nitroprusside (SNP).ResultsComplex I+II, state 3 respiration was significantly lower in the old (young: 12.13±0.48, old: 8.41±0.68 pmol/s/mg), while state 4 was significantly higher in the old (young: 4.69±0.2, old: 7.59±0.67 pmol/s/mg). The respiratory control ratio (RCR), state 3/state 4 respiration, was also significantly attenuated in the old (young: 2.6±0.17, old: 1.19±0.09). Endothelium‐dependent vasodilation was attenuated in the old whether induced by flow (young: 103±6, old: 46±5 %; P<0.01) or ACh (young: 110±12, old: 53±4 %; P<0.01), while endothelial‐independent vasodilation was not altered by age. Thus, there was significant correlation between state 3 respiration and flow‐mediated (r=0.61, P<0.01) and ACh‐dependent vasodilation (r=0.62, P<0.01), but not with the SNP‐dependent response. State 4 respiration was negatively correlated only with ACh‐dependent vasodilation (r=−0.45, P<0.05). RCR also exhibited a strong positive correlation with flow‐mediated (r=0.77, P<0.001) and ACh‐dependent vasodilation (r=0.81, P<0.001), but not the SNP‐dependent response.ConclusionsAdvancing age results in altered mitochondrial respiratory capacity and these changes in mitochondrial function are strongly linked to age‐related endothelial dysfunction in SMFAs.Support or Funding InformationThe National Heart, Lung, and Blood Institute at the National Institute of Health (PO1 HL1091830) and the Veteran's Administration Rehabilitation Research and Development Service (E6910‐R, E1697‐R, E1433‐P, E9275‐L and E1572‐P).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identifying Pathways for Apolipoprotein A2 Actions on Respiratory Rhythmogenesis
Authors: Strohl Kingman Perkins , Azzam Sausan , Saleh Dania 
Published: June 2021

ObjectivesApolipoprotein A2 (ApoA2) appears has a deterministic role on respiratory stability in the mouse (Gillombardo et al 2017). One roadblock to understanding its brain mRNA and protein effects is a lack of fundamental information on how this HDL‐cholesterol factor might act in the CNS. To address this gap, here we utilized a label‐free proteomics approach and conducted a brain proteome analysis to define protein perturbations and dysregulated pathways comparing ApoA2 presence (Apoa2‐WT) or absence (Apoa2‐KO).MethodsEight WT and 8 KO provided desanguinated CNS samples which were lysed (FASPed), and subjected to trypsin digestion. Each sample was analyzed by LC‐MS/MS via LTQ Orbitrap Elite mass spectrometer. Differential quantification of peptides was performed using Rosetta Elucidator. Peptide and protein assignments were made using peptide and protein teller algorithms (FDR <2%). ANOVA was performed comparing peptide intensities across groups. Protein fold changes were determined by dividing the average protein intensity for each peptide in each group. Significant proteins (P‐value ≤ 0.05 and fold change (FC) with ≤‐2 and ≥+2) were imported into Crosstalker for network and pathway analyses.ResultsOur quantitative proteomic analysis identified and quantified a total of 6307 peptides of which, 2995 peptides were statistically significant at p 0.05 using one‐way ANOVA. Of those, 1150 peptides mapping to 298 unique proteins were differentially expressed across groups based on two cutoff criteria: (1) two or more peptides (P ≤0.05) per protein, and (2) each peptide has a fold change of ±2 or greater. Network and pathway analyses of the differentially expressed proteins identified significant pathways (P ≤0.05) associated with the ApoA2 presence or absence. Notable alterations were peptides related to mitochondrial function and metabolism (citric acid cycle and respiratory electron transport, integration of energy metabolism, mitochondrial protein import, mitochondrial biogenesis), RNA, amino acids and carbohydrate metabolism, as well as neuronal signaling.ConclusionsOur work identified altered peptide distributions representing proteins affecting pathways in energy homeostasis, cellular bioenergetics, and neuronal (neurotransmitters) systems related to Apoa2 loss at a cellular level.Support or Funding InformationSupported by VA Research Serve Merit Award and the NIHThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Similar alterations of DNA oxidation levels following prolonged exercise in adolescent boys and girls matched for aerobic capacity
Authors: Yasuda Nobuo , Tanioka Toshihiro 
Published: June 2021

The purpose of this study was to investigate effects of endurance exercise on oxidative DNA damage in adolescent boys and girls matched for aerobic capacity. Aerobically trained adolescent boys (B; n = 15, age: 15.1 ± 1.7 years, height: 165.8 ± 6.9 cm, body weight: 58.4 ± 12.1 kg, body mass index: 21.1 ± 3.2 kg/m2, body fat: 13.1 ± 6.0 %, peak oxygen uptake relative to fat free mass: 77.1 ± 8.5 VO2peak/kg FFM/min) and girls (G; n = 14, age: 14.6 ± 1.0 year, height: 156.3 ± 3.1 cm, body weight: 52.1 ± 4.3 kg, body mass index: 21.3 ± 1.6 kg/m2, body fat: 23.7 ± 5.1 %, peak oxygen uptake relative to fat free mass: 71.8 ± 6.1 VO2peak/kg FFM/min [mean±SD]) matched for peak oxygen uptake relative to fat free mass were enrolled as the participants. Urinary samples were taken at the 2‐hour period before (pre) and after (immediately post and post 24–26 hours) 60 min of cycling exercise at 65%VO2peak, followed by the analysis of urinary 8‐hydroxy‐2′‐deoxyguanosine (8‐OHdG: a potential biomarker of whole body DNA damage and repair) determined with high performance liquid chromatography with electrochemical detection. Two‐way (time × sex) analysis of variances demonstrated no significant main effects for time, sex or interaction (Pre: 3.7 ± 1.8, Post: 4.2 ± 1.9, Post 24–26h: 3.9 ± 1.4 for B; Pre: 3.8 ± 0.9, Post: 4.1 ± 1.3, Post 24–26h: 3.6 ± 0.9 ng/mg creatinine for G) regarding urinary 8‐hydroxy‐2′‐deoxyguanosine level. These results of the present study suggest that no detrimental DNA damage can be observed after prolonged moderate exercise in aerobically trained boys and girls, potentially because of the enhanced antioxidant defense responses. Furthermore, the pubertal boys and girls appear to have similar DNA oxidation responses at the whole‐body level when normalized to peak oxygen uptake relative to fat free mass.Support or Funding InformationSupported partly by funds from the Grant‐in‐Aid for Scientific Research (C) (Grant No.16K01606) and Futaba Electronics Memorial Research Foundation (2016–29) in Japan.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exercise Training Improves Metaboreflex Control of Sympathetic Nerve Activity in Obstructive Sleep Apnea
Authors: Guerra Renan Segalla , Goya Thiago T. , Silva Rosyvaldo F. , Barbosa Eline R.F. , Lima Marta F. , Lorenzi‐Filho Geraldo , Negrão Carlos Eduardo , Ueno‐Pardi Linda Massako 
Published: June 2021

BACKGROUNDWe have recently demonstrated that patients with obstructive sleep apnea(OSA) have abnormal muscle metaboreflex control of muscle sympathetic nerve activity(MSNA).OBJECTIVETo test the hypothesis that exercise training (ET) would improve muscle metaboreflex control of MSNA in patients with OSA.METHODS41 individuals with OSA(polysomnography), age = 51±1 years, BMI = 29.5±0.6 kg/m2, were randomly divided into untrained (n=21), and trained (n=20) groups. Heart rate, blood pressure, and MSNA(microneurography) were continuously measured during 4 min at rest, 3 min of handgrip static exercise (30% maximal voluntary contraction), and 2 min post handgrip muscle ischemia(PHMI). The metaborreflex sensitivity was evaluated as the difference in MSNA between first and second min PHMI and resting period. ET program was conducted for six months, three session/week. Each session consisted of 45 min of cycle ergometer, 10 min of local strengthening exercise, and 5 min of cool down.RESULTSBaseline age, BMI, exercise capacity, and hemodynamic variables were similar between groups. Likewise, the exercise and PHMI responses were not different between groups. Of note, MSNA returned to baseline during the PHMI period in both groups, which is an indicative of impaired muscle metaboreflex control. ET significantly increased exercise capacity. ET reduced MSNA levels at rest (36±3 vs. 25± 2 bursts/min), and during exercise (43±4 vs. 32± 3 bursts/min). More interestingly, MSNA remained elevated during PHMI in comparison to rest period in the exercise‐trained individuals (29±2 vs. 25± 2 bursts/min). No change in the untrained patients was found.CONCLUSIONSET decreases sympathetic nerve activity during moderate handgrip exercise and improves muscle metaboreflex control of MSNA in patients with OSA.Supported by FAPESP # 2010/15064‐6Support or Funding InformationFAPESP # 2010/15064‐6This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.