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Citations Search

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14, 1–12. Analysis of sensitivity and cell death pathways mediated by anti-cancer drugs using three-dimensional culture system. 2018

Kinoshita, T., Higuchi, H., Niibe, A.-K., Sakai, G., Hamamoto, Y., Takaishi, H. and Kanai, T.

Notes: 3D spheroid cultures were used to compare the anti-cancer cell effects between conventional chemotherapeutic drugs and selected multi-kinase inhibitors. Caspase-Glo® 3/7 Assay was used to study cell death. (4996) (4996)

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Toxicol. Lett. 291, 138-148. Cytotoxicity of 34 FDA approved small-molecule kinase inhibitors in primary rat and human hepatocytes. 2018

Zhang J., Ren, L., Yang, X., White, M., Greenhaw, J., Harris, T., Wu, Q., Bryant, M., Papoian, T., Mattes, W., and Shi, Q.

Notes: These authors used the Caspase-Glo® 3/7 and CellTiter-Glo® Assays in studies to determine the cytotoxicity of  34 small molecule kinase inhibitors in primary rat and human hepatocytes. (5009)

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Toxicol. Appl. Pharmacol. 348, 1-13.. Doxorubicin triggers bioenergetic failure and p53 activation in mouse stem cell-derived cardiomyocytes.

  2018

Cunha-Oliveira, T., Ferreira, L.L., Coelho, A.R., Deus, C.M., and Oliveira, P.J.

 

Notes: These authors used cultured induced-pluripotent stem cell-derived mouse cardiomyocytes to investigate the effects of Doxorubicin on cell and mitochondrial metabolism and on stress responses.They used the Caspase-Glo® 3/7  and Caspase-Glo®-9 Assays to detect the respective caspase activities, and the CellTiter-Glo® Assay to determine cell viability. (5008)

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Oncogene 37(1), 39-51. Nuclear factor E2-related factor-2 has a differential impact on MCT1 and MCT4 lactate carrier expression in colonic epithelial cells: a condition favoring metabolic symbiosis between colorectal cancer and stromal cells. 2018

Diehl, K., Dinges, L.A., Helm, O., Ammar, N., Plundrich, D., Arlt, A., Röcken, C., Sebens S., and Schäfer, H.

Notes: These authors studied the role of the antioxidant transcription factor nuclear factor E2-related factor-2 (Nrf2) in adaptation to inflammatory/environmental stress in malignant colonic epithelial cells. They used the Dual-Glo® Luciferase Assay and pGL3-Basic Vector in reporter assays, and the CellTiter® 96, Caspase-Glo® 3/7 and Glucose Uptake-Glo™ Assays to investigate Nrf2 effects on cell viability and metabolism. They found that Nrf2 has an impact on the metabolism in premalignant colonic epithelial cells exposed to inflammatory M1 macrophages, an effect accompanied by growth and survival alterations.  (5014)

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Cancer Res. 77, 4102-15. The alkylating chemotherapeutic temozolomide induces metabolic stress in IDH1-mutant cancers and potentiates NAD+ depletion-mediated cytotoxicity 2017

Tateishi, K., Higuchi, F., Miller, J.J., Koerner, M.V.A., Lelic, N., Shankar, G.M., Tanaka, S., Fisher, D.E., Batchelor, T.T., Iafrate, A.J., Wakimoto, H., Chi, A.S., Cahil, D.P.

Notes: CellTiter-Glo was used to assess cell viability. Caspase-3/7 activities were tested by Caspase-Glo 3/7 Assay. NAD/NADH-Glo and NADP/NADPH-Glo were used to quantify NAD+, NADH, NADP+, and NADPH. (4991)

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Nature 543(7644), 205-210. Autophagy maintains the metabolism and function of young and old stem cells. 2017

Ho, T.T., Warr, M.R., Adelman, E.R., Lansinger, O.M., Flach, J., Verovskaya, E.V., Figueroa, M.E., and Passegué, E.

Notes: These authors investigated how autophagy controls haematopoietic stem cell (HSC) function, and how changes in autophagy levels affect HSC ageing. They showed that loss of autophagy in HSCs causes accumulation of mitochondria and an activated metabolic state. They used the Caspase-Glo® and CellTiter-Glo Assays to detect apoptosis and ATP, respectively, and used the Glucose Uptake-Glo™ Assay to study metabolic changes. (5018)

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Cancer Res. 77, 4102–15. The alkylating chemotherapeutic temozolomide induces metabolic stress in IDH1-mutant cancers and potentiates NAD+ depletion-mediated cytotoxicity. 2017

Tateishi, K., Higuchi, F., Miller, J.J., Koerner, M.V.A., Lelic, N., Shankar, G.M., Tanaka, S., Fisher, D.E., Batchelor, T.T., Iafrate, A.J., Wakimoto, H., Chi, A.S. and Cahill, D.P.

Notes: Cell viability of IDH1 wild-type or mutant cell lines under treatment conditions was assessed using the CellTiter-Glo® Luminescent Cell Viability Assay. From these numbers, the IC50 values were calculated. To evaluate apoptosis, 7,000–8,000 cells were treated with DMSO or compounds singly or in combination (12.5nmol/L NAMPT inhibitors and/or 200μmol/L temozolomide or 50μmol/L Z-VAD-FMK) for 6 or 24 hours and then measured using Caspase-Glo® 3/7 Assay. Changes in NAD+, NADH, NADP+, and NADPH were assessed using the NAD/NADH-Glo™ and NADP/NADPH-Glo™ Assays. (4946)

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Cell Signal 28, 284–93. Elongation factor 2 kinase promotes cell survival by inhibiting protein synthesis without inducing autophagy. 2016

Moore, C.E.J., Wang, X., Xie, J., Pickford, J., Barron, J., Regufe da Mota, S., Versele, M. and Proud, C.G.

Notes: Inhibition of HCT116 cell eEF2 kinase with the JAN-849 inhibitor lead to a dramatic increase in caspase-3/7 activity when the cells were cultured in medium without glucose for 24 hours. The increase could be blocked with cycloheximide (translation elongation inhibitor) or harringtonine (protein synthesis initiation inhibitor). Caspase activity was monitored with the Caspase-Glo® 3/7 Assay. A549 cells exposed to glucose-free medium had an increase in cytotoxicity that was partially abrogated by cycloheximide treatment. Knockdown of eEF2 kinase expression with siRNA treatment doubled the amount of cell death in glucose-free media, and cycloheximide treatment reduced cell death to about half. Cytotoxicity was monitored with the CellTox™ Green Cytotoxicity Assay using the end-point protocol. (4709)

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Clin. Can. Res. 22, 4452-65. Myc-driven glycolysis is a therapeutic target in glioblastoma.  2016

Tateishi, K., Iafrate, A.J., Ho, Q., Curry, W.T., Batchelor, T.T., Flaherty, K.T., Onozato, M.L., Lelic, N., Sundaram, S., Cahill, D.P., Chi, A.S. and Wakimoto, H.

Notes: CellTiter-Glo® Cell Viability Assay was used to assess the vulnerability of myc-driven cancers to inhibitors of the NAMPT. Sensitivity was correlated to the level of myc expression. Apoptosis was assessed with the Caspase-Glo® 3/7 Assay. The NAD/NADH-Glo® Assay was used to provide a quantitative measure of NAD+ in the cells under study. (4835)

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Cell Death Dis. 7, e2073. X-linked inhibitor of apoptosis protein mediates tumor cell resistance to antibody-dependent cellular cytotoxicity 2016

Evans, M.K., Sauer, S.J., Nath, S., Robinson, T.J., Morse, M.A. and Devi, G.R.

Notes: The authors demonstrate that the X-linked inhibitor of apoptosis (XIAP) drives anti-EGFR and anti-HER antibody resistance in inflammatory breast cancer both through its effect on caspase and suppression of ROS accumulation. They use the CytoTox ONE Homogeneous Membrane Integrity Assay to assess cytotoxicity in wild type SUM149 and SUM190 cells, and  rSUM149 and rSUM190 cells (cell lines that have acquired resistance to treatment agents) upon treatment with cytotoxic agents. The Caspase-Glo® 3/7 Assay was used to assess apoptosis in wild type cells, rSUM149 and in cells over expressing XIAP (treated and untreated). (4726)

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Tumor Biol. , ePub ahead of Print Aug 5. Approach for chemosensitization of cisplatin-resistant ovarian cancer by cucurbitacin B 2015

El-Senduny, F.F., Badria, F.A., EL-Waseef, A.M., Chauha, S.C. and Halaweish

Notes: The authors of this study sought to determine whether cucurbitacin B has antitumor activity against the ovarian cancer cell line A2870 and whether it can sensitize the cisplatin-resistant cell line A2780CP to treatment. They compared caspase activity in A2780CP cells either preincubated with cucurbitacin B before treatment with cisplatin or cells not pretreated using the Caspase-Glo® 3/7 Caspase Assay. Oxidized and total glutathione were measured from both cell lines (before and after cisplatin exposure, with and without preincubation with cucurbitacin B) using the GSH/GSSH-Glo™ Glo Assay. The level of reactive oxygen species in cell lines was also measured by detecting ROS converted to H2O2 using the ROS-Glo™ Assay. Cell Viabilty of 3D spheroids formed from the A2780CP cell line was assessed using the CellTiter-Glo® 3D Cell Viability Assay. (4581)

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Assay Drug Dev. Technol. 13, 456–65. Bioluminescent, nonlytic, real-time cell viability assay and use in inhibitor screening 2015

Duellman, S.J., Zhou, W., Meisenheimer, P., Vidugiris, G., Cali, J., Gautam, P., Wennerberg, K. and Vidugiriene, J.

Notes: The authors describe a homogeneous, nonlytic, bioluminescent assay that measures cell viability in real time. They monitored cell health for 72 hours from the same test samples, distinguished differential cell growth, and investigated drug mechanism of action by analyzing time- and dose-dependent drug effects. The real-time measurements enabled them to detect cell death immediately (>75% signal decrease within 15 minutes of digitonin addition), analyze drug potency versus efficacy, and identify cytostatic versus toxic drug effects.They then screened an oncology compound library (Z′ = 0.7) and identified compounds with varying activity at different time points. (4590)

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Br. J. Cancer 112, 1536–45. Low-temperature plasma treatment induces DNA damage leading to necrotic cell death in primary prostate epithelial cells. 2015

Hirst, A.M., Simms, M.S., Mann, V.M., Maitland, N.J., O’Connell, D. and Frame, F.M.

Notes: Low-temperature atmospheric pressure plasmas (LTP) were examined for the mechanism of toxicity on cultured immortalized and primary prostate cancer cells. LTP induces an increase in H2O2 in the culture media as judged by the ROS-Glo™ H2O2 Assay leading to cell death as measured with the CellTox™ Green Cytotoxicity Assay. CellTox™ Green was quantified with a plate reader and verified by fluorescent microscopy. The cells experienced no caspase-3/7 activation as judged through use of the Caspase-Glo® 3/7 Assay, leading to the conclusion of necrotic cell death. (4711)

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PLos ONE 10, e0129058. New small molecules targeting apoptosis and cell viability in osteosarcoma.  2015

Maugg, D. et al.

Notes: A phenotypic screen of a 25,000 compound library for inhibitors of U2OS osteosarcoma cell line growth identified two compounds that showed activity of osteosarcomas and not HepG2 or primary human osteoblasts.  Cell Viability in the primary and secondary screens was monitored with the CellTiter-Blue® Cell Viability Assay.  Subsequent studies identified the mechanism of cell death as apoptosis as judged by the Caspase-Glo® 3/7 Assay and cytotoxicity measured with the CellTox™ Green Cytotoxicity Assay.
(4649)

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Biochem. Biophys. Res. Commun. 446, 30-6. Reversine increases the plasticity of lineage-committed preadipocytes to osteogenesis by inhibiting adipogenesis through induction of TGF-β pathway in vitro. 2014

Park, J.G., Lee, D.-H., Moon, Y.S., and Kim, K.-H.

Notes: 3T3-L1 preadipocytes were treated with various levels of reversine and monitored for induction of apoptosis with the Caspase-Glo® 3/7 Assay with data collected on a GloMax® Instrument. Changes in gene expression of three targets upon reversine treatment were examined through total RNA isolation with the ReliaPrep™ RNA Cell Miniprep System, reverse transcription with the ImProm-II™ Reverse Transcription System followed by dye-based qPCR analysis. (4596)

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Biochem. Pharmacol. October 24, epub ahead of print. Ibandronate  increases the expression of the pro-apoptotic gene FAS by epigenetic mechanisms in tumor cells. 2012

Thaler, R., Spitzer, S., Karlic, H., Berger, C., Klaushofer, K. and Varga, F.

Notes: Caspase-Glo® 3/7 and Caspase-Glo® 8 Assays were used to assess activation of apoptosis pathways in MC3T3-E1 cells (clonal mouse), U-2 OS human osteoscarcoma cell line and CCL-51 cells (mouse mammary gland tumor cells). Although ibandronate reduced cell proliferation in all cell lines, its effect on activation of caspases was different in neoplastic versus non-neoplastic cells. Caspase-8 and caspase-3/7 activities were reduced in MC3T3-E1 cells after 72 hours treatment with ibandronate. In two tumor cell lines assayed, opposite results were seen: caspase-8 and caspase-3/7 activities increased in U-2 OS cells and in CCL-51 cells. Luminescence was detected using a GloMax® 96 Microplate Luminometer. (Figure 1 in the paper)

To analyze FAS promoter methylation levels, fragments of the targeted promoter regions were generated by digestion of genomic DNA using CpG methylation insensitive restriction enzymes MboII and PstI from cells cultured in the presence of ibandronate for varying lengths of time. The authors demonstrated that FAS promoter methylation is altered in tumor cells in the presence of ibandronate. (4253)

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J. Biol. Chem. 287, 14052–68. MDM2 protein-mediated ubiquitination of NUMB protein: Identification of a second physiological substrate of MDM2 that employs a dual-site docking mechanism. 2012

Sczaniecka, M., Gladstone, K., Pettersson, S., McLaren, L., Huart, A.S. and Wallace, M.

Notes: The E3 ubiquitin ligase murine double minute 2 (MDM2) regulates transcriptional activity and expression levels of the p53 tumor suppressor protein. In this article, the authors show that the cell fate determinant NUMB interacts with MDM2, disrupting the interaction of MDM2 and p53 and preventing the ubiquitination of p53. To study protein-protein interactions between NUMB and MDM2, the authors used the pFN2A (GST) Flexi® Vector to express full-length and partial NUMB sequences as GST fusion proteins for protein pull-down experiments. To examine their hypothesis that the interaction between NUMB and MDM2 increases p53 levels and thus apoptosis in cells, they treated ZR75 human breast carcinoma cells and A375 human melanoma cells with NUMB-derived peptides and small molecule ligands that block the interaction between NUMB and MDM2. These treatments resulted in increased levels of p53 protein, Annexin V staining as well as caspase-3/7 activity, as determined using the Caspase-Glo® 3/7 Assay System. (4351)

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PLos ONE 6(6), e20994. Antimetastatic Effects of Phyllanthus on Human Lung (A549) and Breast (MCF-7) Cancer Cell Lines. 2011

Lee S.H., Jaganath I.B., Wang S.M., and Sekaran, S.D.

Notes: These authors investigated the ability of Phyllanthus plant extracts to affect the metastatic activity of human lung (A549) and breast (MCF-7) cancer cell lines. They initially used CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay and the GloMax®-Multi Detection System absorbance module to determine cytotoxicity of various Phyllanthus extracts. After determining the effective dose, the authors investigated the ability of these plant compounds to inhibit/reduce metastatic activity. They then evaluated the mechanism of cell death in treated cells using the Caspase-Glo® 3/7 Assay and the Glomax® Multi Detection System luminescence module to measure caspase activity, and the CytoTox-ONE™ Homogeneous Membrane Integrity Assay to measure LDH activity. (4196)

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Blood 117(1), 352-61. IL-6 in human cytomegalovirus secretome promotes angiogenesis and survival of endothelial cells through the stimulation of survivin. 2011

Botto, S., Streblow, D.N., DeFilippis, V.,  White, L., Kreklywich, C.N., Smith, P.P., Caposio, P.

Notes: In this study, caspase-3/7 activity in HUVECs was measured using the Caspase-Glo® 3/7 Assay. Luminescence was measured with the GloMax®-96 Microplate Luminometer. (4207)

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J. Biol. Chem. 286, 21546–21554. TWEAK induces apoptosis through a death-signaling complex comprising Receptor-interacting protein 1 (RIP1), Fas-associated death domain (FADD), and caspase-8. 2011

Ikner, A. and Ashkenazi, A.

Notes: The authors of this study set out to describe the mechanism of cell death through which TNF-like weak inducer of apoptosis (TWEAK) exerts its apoptotic effect on certain cancer cells. The used the CellTiter-Glo® Cell Viability Assay and the Caspase-Glo® 3/7 Assay to investigate cell viability and mechanism of cell death in HSC3 cells treated with TWEAK. They looked at caspase-8 activity in cells treated with TWEAK in the presence or absence of a caspase-8 inhibitor using the Caspase-Glo® 8 Assay. They showed that TWEAK induces caspase-dependent apoptosis in these cells. (4170)

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Biochem. Pharmacol. epub ahead of print. Identification of known drugs that act as inhibitors of NF-kappaB signaling and their mechanism of action 2010

Miller SC, Huang R, Sakamuru S, Shukla SJ, Attene-Ramos MS, Shinn P, Van Leer D, Leister W, Austin CP, Xia M.

Notes: Dysregulation of the NF-kB pathway has been associated with the formation of a wide variety of tumors and other cancers, as well as diseases, including chronic inflammation and immunodeficiency. Because of the association of constitutive NF-kB regulation and tumors, inhibition of the NF-kB pathway by small molecule antagonists was thought to be a means of reversing or halting the growth and spread of tumors. The authors screened compounds from a database (the NCGC Pharmaceutical Collection or NPC) of small molecule compounds: 52% of the compounds have been approved for human or animal use by the FDA, 22% were drugs approved for use in Europe, and another 25% either drugs approved in other countries or compounds that have been tested in clinical trials. The database served as a source from which to rapidly and efficiently identify already approved drugs that inhibited NF-kB. They used a quantitative high-throughput screening format. To identify small molecules that could inhibit the NF-kB pathway, the compounds were initially screened using a cell-based NF-kB lactamase reporter gene assay, with TNFalpha and MG132 as positive controls. (TNFalpha induced NF-kB coupled beta-lactamase activity, while MG132 blocked TNFalpha induction NF-kB-coupled beta-lactamase activity.) After several rounds of screening, 20 compounds were further studied for their NF-kB inhibition, with NF-kB luc2P HEK293 cells. After a concordance rate of 95% between the luciferase and beta-lactamase tests, compounds were additionally examined for their ability to affect caspase 3/7 activity, for the ability to disrupt the electrochemical gradient across the mitochondrial membrane in relation to cellular apoptosis, as well as tests of the inhibitors on cancer cell viability and affects on LDH release, an indicator of cell necrosis.
(4049)

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Current Chemical Genomics 3, 33-41. In vitro viability and cytotoxicity testing and same-well multi-parametric combinations for high-throughput screening 2009

Niles, A.L., Moravec, R.A. and Riss, T.L.

Notes: The authors review the use of in vitro cytotoxicity testing in drug discovery to characterize the toxic potential of new chemical entities (nce) at the earliest stages of profiling. DOI: 10.2174/1875397300903010033 (4000)

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Toxicology in Vitro 23, 1170-1171. Multiplexed assay panel of cytotoxicity in HK-2 cells for detection of renal proximal tubule injury potential of compounds 2009

Wu, Y., Connors, D., Barber, L., Jayachandra, S., Hanumegowda, U.M. and Adams, S.P.

Notes: The authors describe a multiplexed in vitro assay to detect nephrotoxicity and gain information about mechanism of cell death in HK-2 (human kidney-2) cells. The multiplexed assay involved an LDH assay to detect necrosis, a caspase-3/7 assay to detect apoptosis, a reazurin assay to assess metabolic state, and a DNA dye staining assay to monitor nuclear morphology. (4002)

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Expert Opin. Drug Metab. Toxicol. 4, 103–120. Bioluminescent assays for ADMET 2008

Cali, J.J., Niles, A., Valley, M.P., O’Brien, M.A., Riss, T.L., and Shultz, J.

Notes: The authors of this review article highlight the use of bioluminescence as a readout for high-throughput ADME/Tox assays. They discuss three strategies for designing bioluminescent assays, using either luciferase, ATP or luciferin substrates as the limiting reagents for a luciferase-catalyzed reaction. Reporter gene assays limit the production of luciferase by tying it to a promoter or DNA regulatory region of interest. Such assays can be used to study genes that are regulated by drugs and other xenobiotics. Bioluminescent assays in which ATP is the limiting reagent of the luciferase reaction can be designed to monitor cell viability or the activity kinases. Bioluminescent assays in which the substrate is limiting can be designed so that the activity of a particular enzyme results in the production of a luciferin substrate that can, in turn, be acted upon by luciferase. (3926)

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Blood 111, 33498-33506. Transcription factor Erg regulates angiogenesis and endothelial apoptosis through VE-cadherin. 2008

Birdsey GM, Dryden NH, Amsellem V, Gebhardt F, Sahnan K, Haskard DO, Dejana E, Mason JC, Randi AM.

Notes: These authors showed that the ETS transcription factor Erg interacts with the VE-cadherin promoter region and regulates endothelial apoptosis through this interaction. They demonstrated that inhibition of Erg by siRNA resulted in decreased VE-cadherin mRNA and protein levels, and showed that Erg interacts with the VE-cadherin promoter using a CHIP assay. To show the functional relevance of this interaction, HeLa cells were transfected with a pGL4 Vector containing the VE-cadherin promoter region and an expression vector containing Erg2 cDNA. In this reporter assay, Erg overexpression resulted in ~1.8 fold transactivation of VE-cadherin promoter activity, as measured using the Dual-Luciferase® Reporter Assay System. Inhibition of Erg in human umbilical vein endothelial cells also resulted in a loss of viability and an increase in activation of caspase 3 and caspase 7. The authors showed that apoptosis could be significantly decreased by overexpression of VE-cadherin, indicating that Erg regulates survival partially via its interaction with VE-cadherin. The Caspase-Glo® 3/7 Assay was used to measure caspase activity in these experiments. (3872)

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