bluuebunny/crossref_metadata_2025_split huggingface dataset:
Curcuminoids Supress Cell Proliferation of Glioblastoma by Down‐regulation of CXCR4 Signaling Pathway
Authors: Shivachar Amruthesh C. , Nnamdiebere Joseph Ekeada , Jordan Bryan C. , Selvam Chelliah , Mathews Omana P , Ranganna Kasturi 
Published: November 2023

Glioblastoma multiforme is one of the glial‐derived malignant primary brain tumors in adults with a poor prognosis. The current average survival period still remains between 14–16 months despite various treatment modalities. The robust tumor progression is mainly attributed to its pro‐angiogenic and anti‐hypoxic targets that many current chemotherapeutics may miss. Curcumin (#1) is a plant derived dietary constituent, and has been shown to play a role in preventing many cancers. Here we investigated for the first time the potencies and the molecular mechanisms of action of #1 and its three pyrazole analogs (#7, #20, and #32) that were prepared through an acid catalyzed condensation with various hydrazine derivatives. Heterocyclic pyrazole modification introduces conformational constraint that is beneficial for improving potency and in vivo chemical stability for curcumin analogs. Cultures of grade IV glioblastoma multiforme (GBM) cells (10,000 cells/cm2) in 24‐well plates were treated with various concentrations (0.01–10micrMole) of #1, #7, #20 or #32 for 24 hr. along with vehicle controls. Cell viability/toxicity was assessed using the fluorescent dyes Hoechst 33342 (1 mg/ml, 20 min at 37°C), which stains DNA of both living and dead cells and propidium iodide (PI; 5 mg/ml, 5 min, 37°C), which stains the DNA of cells with disrupted membranes. The cell death was calculated [pink cells × 100/total cells] from 3–5 non‐overlapping microscopic fields in an Olympus IX81 inverted fluorescent microscope: dead cells are double stained (pink), while viable ones remained blue and were quantified using Prism Graph Pad software. The cell proliferation was measured by the uptake of BrDU by immunofluorescent method. In the sister cultures, the effect of these analogs on the expression of chemokine receptor 4 (CXCR4) that is known to affect tumor progression and metastasis in these cells was measured by western blotting analyses. Our results show that the tested analogs dose‐dependently decreased cell viability and inhibited cell proliferation in the order #32> #20> #7> #1, suggesting that heterocyclic pyrazole modification increased the potency of the parent compound #1. Additionally, we noted that #1 and its pyrazole analogs significantly down‐regulated the expression of CXCR4 over vehicle‐treated controls. Thus our preliminary results demonstrate that curcuminoids may suppress cell proliferation of GBM by down‐regulation of CXCR4 signaling pathway.Support or Funding InformationThis work was supported, in part, by an Institutional seed grant (ACS), and by NIH‐RCMI‐Program Molecular Biology Core Facility (KR).
STK11 Loss Stabilizes MCL1 and Confers Targetable Drug Resistance in KRAS Mutant NSCLC
Authors: Kahle Michael , Akbay Esra , Christensen Camilla , Sholl Lynette , Becker Jeffrey , Soucheray Margaret , Chen Zhao , Pulido Ines , Kikuchi Eiki , Gao Yandi , Dennis Peter , Qiu Wei , Al‐Shahrour Fatima , Johnson Neil , Awad Mark , Carretero Julian , Wong Kwok‐Kin , Shimamura Takeshi 
Published: November 2023

The combined KRAS/STK11 (serine‐threonine kinase 11, LKB1) mutated genotype is prevalent and predictive of poor clinical outcome in NSCLC as tumors are drug resistant. Here, we demonstrate that the bromodomain and extraterminal inhibitor (BETi) JQ1 can induce apoptosis in KRASm NSCLC by targeting the pro‐apoptotic BIM anti‐apoptotic BCL2/BCL‐XL axis, but not in KRASm/STK11m due to additional elevated MCL1. Specifically, loss of STK11 stabilizes anti‐apoptotic MCL1 protein via activation of the PI3K‐mTOR signaling axis. In NSCLC cell line models, we have demonstrated that overexpressing myristoylated‐PI3K increased MCL1 expression whereas the PI3K/mTOR inhibitor BEZ235 reduced MCL1 in a dose, time, and proteasome‐dependent, yet relatively GSK3β‐independent, manner. Furthermore, MCL1 stability also attenuates the antitumor efficacy of the direct BCL2/BCL‐XL inhibitor, ABT‐263 (Navitoclax) in KRASm/STK11m cells. However, combination of BETi or ABT‐263 with MCL1 protein reduction by siRNA or BEZ235, or direct MCL1 inhibition by the BH3 mimetic, A1210477, could sensitize resistant cells to apoptosis while no monotherapy was effective. Therefore, STK11 mutation or aberrant PI3K/mTOR pathway activations emerge as biomarkers of BET‐ or BCL2/BCL‐XL inhibitor resistance, suggesting patient selection for combined MCL1 inhibition.Support or Funding InformationThis work is supported by research Scholar grant RSG‐14‐229‐01 from the American Cancer Society and United Against Lung Cancer Legacy Program (Lung Cancer Research Foundation). This work is also supported by research fellowship from Sumitomo life social welfare services foundation (E.K), MINECO (SAF2010‐21769), ISCIII (RD12/0036/0045) and Generalitat Valenciana (ACOMP/2013/156 and ACIF/2013/239) (J.C.), and National Institutes of Health grants CA140594, CA122794, CA163896, CA166480 (K.K.W)
Exercise ameliorates endoplasmic reticulum stress‐mediated vascular dysfunction in atherosclerotic mesenteric arteries
Authors: Hong Junyoung , Kim Kwangchan , Park Eunkyung , Lee Jonghae , Park Yoonjung 
Published: November 2023

IntroductionThe endoplasmic reticulum (ER) stress has been known as a disruption of ER homeostasis promoting to an accumulation of misfolded and unfolded protein response (UPR) that is closely associated with cardiovascular diseases including atherosclerosis. The underlying mechanisms for atherosclerotic mesenteric arterial dysfunction, major cause of chronic mesenteric ischemia, is unclear and no study has been conducted to investigate the effect of exercise on mesenteric arterial dysfunction in atherosclerosis.HypothesisWe hypothesized that ER stress aggravates endothelial dysfunction in mesenteric arteries in atherosclerosisMethodsWe used wild type (WT), wild type with exercise training, running on the treadmill for 12 weeks (WT‐EX), apolipoprotein E knockout (ApoE KO) and ApoE KO with exercise training (ApoE KO‐EX) mice. To test mesenteric artery function, endothelium‐dependent (acetylcholine, ACh) vasodilation and endothelium‐independent nitroprusside (SNP) vasodilation of isolated and pressurized mesenteric arteries were measured by a concentration‐dependent manner. To determine underlying mechanism, endothelial nitric oxide synthase (eNOS) inhibitor, (NG‐nitro‐L‐arginine methyl ester (L‐NAME)), ER stress inducer, (tunicamycin), ER stress inhibitor, (Tudca), caspase‐1 inhibitor (AC‐YVARD‐cmk) and uncoupling protein‐2 (UCP‐2) inhibitor, (Genepin) were incubated and ACh‐induced dilation was measured to investigate the underlying mechanisms. To measure the expression of ER stress markers, protein kinase RNA‐like endoplasmic reticulum kinase (PERK) and C/EBPa‐homologous protein (CHOP), in the isolated superior mesentery arteries, immunofluorescence staining was performed.ResultsACh‐induced vasodilation was attenuated in ApoE KO compared to WT but exercise training improved the ACh‐induced vasodilatation in ApoE KO‐EX while endothelium‐independent SNP‐induced vasodilation was not different among the groups. Incubation of Tudca and AC‐YVARD‐cmk improved ACh‐induced vasodilation in ApoE KO. ACh‐induced vasodilation in the presence of L‐NAME, tunicamycin and Genepin were reduced in WT, WT‐EX and ApoE KO‐EX, but not in ApoE KO. The expression of PERK and CHOP in the isolated superior mesentery arteries were increased in ApoE KO compared to WT but exercise training decreased its expression in ApoE KO‐EX compared to ApoE KO.ConclusionOur findings suggest that ER stress plays a significant role in vascular endothelial dysfunction in atherosclerotic mesenteric arteries and exercise restores it through ER stress and its downstream signaling pathways including NOS, UCP‐2 and caspase‐1.
Adenosine receptor dependent regulation of vascular tone in L‐NAME mouse model of hypertension.
Authors: Yadav Vishal R , Teng Bunyen , Mustafa Jamal S 
Published: November 2023

L‐NAME, a nitric oxide synthase inhibitor, down regulates eNOS protein expression and leads to depletion of plasma nitric oxide levels in blood. L‐NAME pre‐treatment causes systemic vasoconstriction, increased vascular resistance and high blood pressure (BP). Thus, L‐NAME model of hypertension is used to study endothelial dysfunction and related vascular effects. Adenosine is an important metabolite, which controls local blood flow through activation of four subtypes of adenosine receptors (ARs). Stimulation of A1 and A3 ARs produce contraction while A2A and A2B ARs cause relaxation in mouse aorta and mesenteric arteries (MAs). In present study, we sought to determine whether ARs play a role in altered vascular tone regulation in this model of hypertension. We used L‐NAME (1 mg/ml) in the drinking water for 4 weeks. The BP was monitored using CODA non‐invasive tail cuff system. For vascular tone studies, aortae and MAs were isolated from control and hypertensive mice. Muscle tension studies were performed using DMT for MAs and organ bath for aorta. Systolic BP recorded at the end of 28 days was (mean ± SE; mmHg) 133.9 ± 3.0 in hypertensive while 107.8 ± 2.6 in control group (n=11). Our DMT data analysis in MAs showed that acetylcholine (Ach) induced relaxation responses were (% PE) 98.5 ± 0.2 and 93.4 ± 1.4 (p<0.05, n=10) in control and L‐NAME hypertensive mice, respectively. Vascular studies in MAs revealed higher CCPA (selective A1AR agonist) induced contractions in hypertensive mice. Particularly, 100 nM CCPA (% PE) produced 24.9 ± 14.1 contraction in control whereas 89.2 ± 27.4 (p<0.05, n=7) in L‐NAME mice. Contrary, NECA (non‐selective AR agonist) induced vascular responses were comparable. Pinacidil (KATP channel opener) induced relaxation was significantly increased in hypertensive mice. Pinacidil (1 μM), produced relaxation of 51.72 ± 9.8 and 76.29 ± 5.9 (p<0.05, n=7) in control and L‐NAME mice, respectively. Phenylephrine induced contraction was reduced to (mg) 278.8 ± 32.8 in L‐NAME mice compared to 492.1 ± 33.8 in control (p<0.05, n=11). Angiotensin (ANG) II induced vascular contractions were reduced to (% KCl) 54.3 ± 9.0 in L‐NAME compared to 122.4 ± 14.0 in control mice (p<0.05, n=7). Our organ bath data in aorta showed that CCPA, NECA, pinacidil PE, KCl and Ach induced responses were comparable in both groups. In conclusion, A1AR induced contraction and KATP channel induced relaxation were both higher in MAs of L‐NAME mice. In addition, ANG II induced contraction were lower in MAs. Thus, suggesting altered responses to ARs and ANG II in L‐NAME induced mouse model of hypertension.Support or Funding InformationSupported by HL 027339
Evaluation of Antiobesity activity of Ethanolic extract of Cashew Apple against high fat diet induced obesity in rodents
Authors: Rani T Jhansy , Prasad K.V.S.R.G. , Dodoala Sujatha 
Published: November 2023

Obesity has doubled since 1980, as per WHO report affecting more than 1900 million adults throughout the world. In recent studies, diets containing antioxidants and branch chain amino acids were demonstrated to have potential effects on fat utilization. Cashew apple juice comprising nutritional components including vitamin C and BCAS reported as an enhancing factor in fat oxidation during exercise and may improve endurance performance (Piyapong Prasertsri et al., 2013). Hence, the current work was planned with an objective to evaluate the anti‐obesity activity of the ethanol extract of cashew apple (ECA). To identify the active constituents, ECA was subjected for analysis using GC‐MS technique. High fat diet induced rodent model was used to induce obesity in the experimental animals. Adult male albino rats were randomized in to five groups (n=6) among which first group was kept on standard rodent chow (normal). The remaining four groups were assigned as obese control, standard & ECA treated groups which received high fat diet along with vehicle, atorvastatin (10 mg/kg, p.o) and ECA (200 & 400 mg/kg, p.o) respectively for 60 days. At the end of study, obese control group animals showed a significant increase in BMI & body wt, serum total cholesterol, triglycerides, LDL‐cholesterol and VLDL‐cholesterol compared to normal group. Administration of ECA (200mg/kg, 400mg/kg) exhibited significant decrease in serum total cholesterol, triglycerides, LDL‐cholesterol and VLDL‐cholesterol levels in ECA treated groups. In addition HDL‐cholesterol levels were also increased distinctly compared to the obese control groups. A notable elevation in the levels of serum creatinine, transaminases & alkaline phosphatase were observed with an intake of high fat diet. These effects were reversed by oral administration of ECA. A significant inhibition of serum malondialdehyde (MDA) and improvement in hepatic enzymatic (catalase, superoxide dismutase), non‐enzymatic(reduced glutathione) levels was also identified as an outcome after treatment with ECA (200mg/kg, 400mg/kg) and standarad (atorvastatin) compared to the obese control groups. Eventually, consumption of cashew apple may even reverse the fatty liver which was quite evident from histology images. GC‐MS analysis revealed the presence of 5‐Hydroxy‐2‐(hydroxymethyl)‐4H‐pyran‐4‐one; 2,3‐dihydro‐3,5‐dihydroxy‐6‐methyl‐4H‐pyran‐4‐one; Hexadecanoic acid; octadecanoic acid; 5‐(Hydroxymethyl)‐2‐furancarboxaldehyde; Furfural; 4‐H‐pyran‐4‐one,3,5, dihydro‐2‐methyl decanoic acid ethyl ester.Based up on the available experimental evidence it can be concluded that ethanolic cashew apple extract possess anti‐obesity activity and reinforce recommending consumption of cashew apple to modulate obesity.Support or Funding Information1. Sri Padmavati Mahila Visvavidyalayam, Tirupati, Andhra Pradesh, India.2. DST, Government of India.
mTORC2 Facilitates the Maturation of Protein Kinase C by a Phosphorylation‐Independent Mechanism
Authors: Baffi Timothy R. , Gould Chistine M. , Wang Zhiyong , Gutkind Silvio , Newton Alexandra C. 
Published: November 2023

Protein Kinase C (PKC) is primed by a series of ordered phosphorylations and conformational transitions to yield an autoinhibited enzyme that is positioned to respond to second messengers. For most PKC isozymes, this maturation to yield a signaling‐competent enzyme depends on the mammalian Target of Rapamycin Complex 2 (mTORC2); however, the molecular mechanisms by which mTORC2 regulates PKC are unknown. Here we use a variety of Förster resonance energy transfer (FRET)‐based live cell imaging assays to examine the role of mTORC2 in the the folding, phosphorylation, and activity of overexpressed PKCbII. We show that in cells lacking Sin1, a critical component of mTORC2, PKC exists in an unprimed, unphosphorylated, and open conformation, with fully‐exposed membrane‐targeting modules. In contrast, overexpression of PKCbII in cells with functional mTORC2 results in accumulation of a fully‐phosphorylated and autoinhibited enzyme. Introduction of Glu at the two phosphorylated residues on the C‐terminal tail is insufficient to bypass the requirement for mTORC2 in the folding of PKC; however, expression of Sin1 in Sin1−/− MEFs rescues the defect, allowing PKC to become phosphorylated and fold into the mature, closed conformation. Furthermore, we demonstrate that PKC is constitutively active in the absence of mTORC2. Finally, analysis of chimeric proteins of an mTORC2‐dependent (PKCbII) and mTORC2‐independent (PKCd) enzyme, coupled with peptide array experiments, identify two critical regions in the kinase domain and C‐terminal tail of PKC that each confer mTORC2‐dependence. Our data support a model in which mTORC2 serves as a molecular chaperone to permit PKC to be processed by phosphorylation, modifications that allow the enzyme to adopt an autoinhibited conformation.Support or Funding InformationThis work was supported by NIH GM43154, UCSD Graduate Training Program in Cellular and Molecular Pharmacology NIH T32 GM007752, and the PhRMA Foundation Pre‐Doctoral Fellowship in Pharmacology/Toxicology.
Exploration of the 5‐HT2A : 5‐HT2C receptor protein complex in vitro
Authors: Felsing Daniel E , Anastasio Noelle C , Gilbertson Scott R , Moeller Gerard F , Cunningham Kathryn A , Allen John A 
Published: November 2023

The disruption of serotonin 5‐HT2A receptor (5‐HT2AR) and 5‐HT2CR function is implicated in psychiatric disorders that range from addiction to schizophrenia. Previous studies have demonstrated that these structurally similar class A G protein‐coupled receptors exist as homomers in living cells (Brea et al., Mol Pharm 75:1380, 2009; Herrick‐Davis et al., Biochemistry 43:13963, 2004), and we are interested in the possibility that these two receptors are in the same protein complex, which may regulate their signaling and/or trafficking. The 5‐HT2AR and 5‐HT2CR co‐localize to neurons in the medial prefrontal cortex (mPFC) of the rat (Nocjar et al, Neuroscience 25:22, 2015) and co‐immunoprecipitate from mPFC preparations (Anastasio et al, ACS Chem. Neurosci. 15:1248, 2015). To expand this work, we designed a luciferase complementation assay (LCA) to test the hypothesis that the 5‐HT2AR and 5‐HT2CR are in close proximity (~50 nm) in living cells. In the LCA, two complementary N‐terminal (NLuc) and C‐terminal (CLuc) components, which alone do not have enzymatic activity, were fused to the 5‐HT2AR and 5‐HT2CR, respectively, and transiently transfected into HEK293 cells. In the presence of the substrate D‐luciferin, we hypothesized that the association of the two receptors would bring the inactive luciferase fragments into close proximity to reconstitute the enzymatic activity of luciferase. The LCA showed robust complementation (i.e., increased luminescence) dependent on cDNA transfection ratios, with optimal signal achieved at a 3:1 ratio of 5‐HT2CR‐NLuc to 5‐HT2AR‐CLuc (n=3, p<0.001). Co‐transfection with plasmid DNA encoding unlabeled 5‐HT2AR or 5‐HT2CR, inhibited the complementation between the two tagged receptors (n=3, p<0.001, and p<0.01, respectively). In ongoing studies we are exploring the effect of receptor selective agonists and antagonists on protein complementation. Taken together, we have detected a 5‐HT2AR:5‐HT2CR complex whose formation is dependent on the relative expression of the individual receptor protomers. Because these receptors are important drug targets in psychiatric disorders, a greater understanding of this novel receptor complex may also provide insights for the development of novel pharmacotherapeutics.Support or Funding InformationNIH NIDA P50 DA033935 and Center for Addiction Research at UTMB
Cross‐tissue telomere length measurement to determine auxiliary marker of brain telomere length in neurosurgery patients
Authors: Lussier Stephanie , Braun Patricia , Potash James , Shinozaki Gen , Stevens Hanna 
Published: November 2023

BackgroundTelomere length, the length of the chromosomal endcaps, is a commonly used biomarker of health and aging, as it may indicate the extent to which cells have undergone damage. Changes in telomere length have been connected to exposure to stress and poor physical health outcomes, such as heart disease, risk of stroke, and coronary artery disease. The use of telomere length as a biomarker, therefore, is reflective of its importance in cellular development and function. As a biomarker in human studies, telomere length is measured in cells taken from blood, saliva, or other easily obtained peripheral cells, even in the study of disorders of the central nervous system (CNS) such as psychiatric illness. It is unclear if telomere length plays a role in pathophysiological processes in the brain. This question could be better addressed by determining if telomere length changes in the CNS are commensurate with those in peripheral tissues.MethodsIn this study, we aim to look at the relationship between CNS neural telomere length measurements in comparison to peripheral measurements in two different tissue types to determine if peripheral telomere length measurements may be proxies for CNS measures. Samples of blood, saliva, and brain were taken in 22 neurosurgery patients before, during, and after surgery. Brain resection surgeries were performed to remove tissue at seizure focal points. Patients had an average age of 32.4 years ranging from 5 to 61 years old and 15 out of 22 patients (68%) were male. Telomere length was measured in all samples using quantitative PCR techniques.ResultsBlood telomere length, but not saliva or brain, negatively correlated with age (r=0.84, p<0.001), a previously reported phenomenon. Comparing pre‐ and post‐surgery samples, we found that saliva telomere length had a trend decrease (paired student's t‐test, p=0.08), while blood telomere length had no change. Intriguingly, we found a trend correlation between brain telomere length and pre‐surgery saliva telomere length (standardized beta = 0.47, r=0.47, p=0.09).ConclusionsWe found that telomere measures in saliva trended to be different before and after surgery, in which a significant intervening variable was use of synthetic glucocorticoids to prevent edema. We hypothesize that exposure to glucocorticoid even on this acute time course may affect saliva telomere length. Further, we have found that there may exist a relationship between brain and pre‐surgery salivary telomere length, but no other examined tissues (post‐surgery saliva, pre‐surgery blood, and post‐surgery blood). These findings may help to identify saliva telomere length as a potential correlate for brain telomere length, which may broaden its applications as a biomarker.Support or Funding InformationUniversity of Iowa CPH Advancing Graduate Student Success Award
Regulation of Cannabinoid‐mediated Neuroprotection by Hydrogen Sulfide in Bovine Isolated Retina
Authors: Mitchell Leah , Robinson Jenaye , Ngele Kalu , Opere Catherine , Ohia Sunny E , Njie‐Mbye Ya Fatou 
Published: November 2023

Hydrogen sulfide (H2S) and endocannabinoids have been reported to possess neuroprotective properties in several mammalian tissues. In the present study, we will investigate the role of H2S in the neuroprotective actions of cannabinoids in the bovine isolated neural retina.MethodsIsolated bovine retinae were exposed to oxidative stress using hydrogen peroxide (H2O2, 100 μM) for 10 minutes. Before exposure to stress, tissues were pretreated with inhibitors of H2S biosynthesis, amino‐oxyacetic acid (AOAA, 30 μM) and α‐keto‐butyric acid (KBA, 1 mM), and the CB1‐receptor antagonist, AM251 (100 nM) prior to treatment with methanandamide (1 nM – 100 nM). Lipid peroxidation was assessed using the Cayman 8‐isoprostane ELISA kit.ResultsIn the presence of H2O2 (100 μM), there was a 20% increase in 8‐isoprostane levels when compared to control. At a concentration of 1 nM, methanandamide significantly (P<0.001) reversed the H2O2 (100 μM)‐induced augmentation in 8‐isoprostane levels in the neural retina. Interestingly, the neuroprotective action caused by methanandamide was abolished by KBA (1 mM) but not by AM251 (100 nM) or AOAA (30 μM).ConclusionsOur results demonstrate that the neuroprotective action of methanadamide can be reversed by an inhibitor of the enzyme, 3‐mercaptopyruvate sulfurtransferase but not by the CB1‐receptor antagonist. Taken together, we conclude that the neuroprotective action of endocannabinoids involve the endogenous biosynthesis of H2S via the cysteine aminotransferase dependent pathway in the bovine retina.