P38 Kinase, SGK1 and NF-κB Dependent Up-Regulation of Na+/Ca2+ Exchanger Expression and Activity Following TGFß1 Treatment of Megakaryocytes
Background: TGFβ1, a decisive regulator of megakaryocyte maturation and platelet formation, has previously been shown to up-regulate both, store operated Ca 2+ entry (SOCE) and Ca 2+ extrusion by Na + /Ca 2+ exchange. The growth factor thus augments the increase of cytosolic Ca 2+ activity ([Ca 2+ ]...
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| Veröffentlicht in: | Cellular physiology and biochemistry 2017, Vol.42 (6), p.2169-2181 |
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| creator | Al-Maghout, Tamer Pelzl, Lisann Sahu, Itishri Sukkar, Basma Hosseinzadeh, Zohreh Gutti, Ravi Laufer, Stefan Voelkl, Jakob Pieske, Burkert Gawaz, Meinrad Lang, Florian |
| description | Background: TGFβ1, a decisive regulator of megakaryocyte maturation and platelet formation, has previously been shown to up-regulate both, store operated Ca 2+ entry (SOCE) and Ca 2+ extrusion by Na + /Ca 2+ exchange. The growth factor thus augments the increase of cytosolic Ca 2+ activity ([Ca 2+ ] i ) following release of Ca 2+ from intracellular stores and accelerates the subsequent decline of [Ca 2+ ] i . The effect on SOCE is dependent on a signaling cascade including p38 kinase, serum & glucocorticoid inducible kinase SGK1, and nuclear factor NFκB. The specific Na + /Ca 2+ exchanger isoforms involved and the signalling regulating the Na + /Ca 2+ exchangers remained, however elusive. The present study explored, whether TGFβ1 influences the expression and function of K + insensitive (NCX) and K + sensitive (NCKX) Na + /Ca 2+ exchangers, and aimed to shed light on the signalling involved. Methods: In human megakaryocytic cells (MEG01) RT-PCR was performed to quantify NCX/NCKX isoform transcript levels, [Ca 2+ ] i was determined by Fura-2 fluorescence, and Na + /Ca 2+ exchanger activity was estimated from the increase of [Ca 2+ ] i following switch from an extracellular solution with 130 or 90 mM Na + and 0 mM Ca 2+ to an extracellular solution with 0 Na + and 2 mM Ca 2+ . K + concentration was 0 mM for analysis of NCX and 40 mM for analysis of NCKX. Results: TGFβ1 (60 ng/ml, 24 h) significantly increased the transcript levels of NCX1, NCKX1, NCKX2 and NCKX5. Moreover, TGFβ1 (60 ng/ml, 24 h) significantly increased the activity of both, NCX and NCKX. The effect of TGFβ1 on NCX and NCKX transcript levels and activity was significantly blunted by p38 kinase inhibitor Skepinone-L (1 µM), the effect on NCX and NCKX activity further by SGK1 inhibitor GSK-650394 (10 µM) and NFκB inhibitor Wogonin (100 µM). Conclusions: TGFβ1 markedly up-regulates transcription of NCX1, NCKX1, NCKX2, and NCKX5 and thus Na + /Ca 2+ exchanger activity, an effect requiring p38 kinase, SGK1 and NFκB. |
| doi_str_mv | 10.1159/000479992 |
| format | Article |
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The growth factor thus augments the increase of cytosolic Ca 2+ activity ([Ca 2+ ] i ) following release of Ca 2+ from intracellular stores and accelerates the subsequent decline of [Ca 2+ ] i . The effect on SOCE is dependent on a signaling cascade including p38 kinase, serum & glucocorticoid inducible kinase SGK1, and nuclear factor NFκB. The specific Na + /Ca 2+ exchanger isoforms involved and the signalling regulating the Na + /Ca 2+ exchangers remained, however elusive. The present study explored, whether TGFβ1 influences the expression and function of K + insensitive (NCX) and K + sensitive (NCKX) Na + /Ca 2+ exchangers, and aimed to shed light on the signalling involved. Methods: In human megakaryocytic cells (MEG01) RT-PCR was performed to quantify NCX/NCKX isoform transcript levels, [Ca 2+ ] i was determined by Fura-2 fluorescence, and Na + /Ca 2+ exchanger activity was estimated from the increase of [Ca 2+ ] i following switch from an extracellular solution with 130 or 90 mM Na + and 0 mM Ca 2+ to an extracellular solution with 0 Na + and 2 mM Ca 2+ . K + concentration was 0 mM for analysis of NCX and 40 mM for analysis of NCKX. Results: TGFβ1 (60 ng/ml, 24 h) significantly increased the transcript levels of NCX1, NCKX1, NCKX2 and NCKX5. Moreover, TGFβ1 (60 ng/ml, 24 h) significantly increased the activity of both, NCX and NCKX. The effect of TGFβ1 on NCX and NCKX transcript levels and activity was significantly blunted by p38 kinase inhibitor Skepinone-L (1 µM), the effect on NCX and NCKX activity further by SGK1 inhibitor GSK-650394 (10 µM) and NFκB inhibitor Wogonin (100 µM). Conclusions: TGFβ1 markedly up-regulates transcription of NCX1, NCKX1, NCKX2, and NCKX5 and thus Na + /Ca 2+ exchanger activity, an effect requiring p38 kinase, SGK1 and NFκB.</description><identifier>ISSN: 1015-8987</identifier><identifier>EISSN: 1421-9778</identifier><identifier>DOI: 10.1159/000479992</identifier><identifier>PMID: 28813704</identifier><language>eng</language><publisher>Basel, Switzerland: Cell Physiol Biochem Press GmbH & Co KG</publisher><subject>Benzoates - pharmacology ; Bridged Bicyclo Compounds, Heterocyclic - pharmacology ; Calcium ; Calcium - metabolism ; Cell Line ; Dibenzocycloheptenes - pharmacology ; Flavanones - pharmacology ; Humans ; Immediate-Early Proteins - antagonists & inhibitors ; Immediate-Early Proteins - genetics ; Immediate-Early Proteins - metabolism ; Megakaryocytes - cytology ; Megakaryocytes - drug effects ; Megakaryocytes - metabolism ; Microscopy, Fluorescence ; NCKX1 ; NCKX2 ; NCKX5 ; NCX1 ; NF-kappa B - antagonists & inhibitors ; NF-kappa B - metabolism ; NF-κB ; Original Paper ; p38 kinase ; p38 Mitogen-Activated Protein Kinases - antagonists & inhibitors ; p38 Mitogen-Activated Protein Kinases - metabolism ; Protein Isoforms - genetics ; Protein Isoforms - metabolism ; Protein-Serine-Threonine Kinases - antagonists & inhibitors ; Protein-Serine-Threonine Kinases - genetics ; Protein-Serine-Threonine Kinases - metabolism ; Real-Time Polymerase Chain Reaction ; SGK1 ; Sodium-Calcium Exchanger - genetics ; Sodium-Calcium Exchanger - metabolism ; Transcription, Genetic - drug effects ; Transforming Growth Factor beta1 - pharmacology ; Up-Regulation - drug effects</subject><ispartof>Cellular physiology and biochemistry, 2017, Vol.42 (6), p.2169-2181</ispartof><rights>2017 The Author(s). Published by S. Karger AG, Basel</rights><rights>2017 The Author(s). Published by S. Karger AG, Basel.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c2802-2227d871e62f134575a6d52593b80811693e619884b26f9242708ce11d7cd3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,864,2100,4022,27634,27922,27923,27924</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28813704$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Al-Maghout, Tamer</creatorcontrib><creatorcontrib>Pelzl, Lisann</creatorcontrib><creatorcontrib>Sahu, Itishri</creatorcontrib><creatorcontrib>Sukkar, Basma</creatorcontrib><creatorcontrib>Hosseinzadeh, Zohreh</creatorcontrib><creatorcontrib>Gutti, Ravi</creatorcontrib><creatorcontrib>Laufer, Stefan</creatorcontrib><creatorcontrib>Voelkl, Jakob</creatorcontrib><creatorcontrib>Pieske, Burkert</creatorcontrib><creatorcontrib>Gawaz, Meinrad</creatorcontrib><creatorcontrib>Lang, Florian</creatorcontrib><title>P38 Kinase, SGK1 and NF-κB Dependent Up-Regulation of Na+/Ca2+ Exchanger Expression and Activity Following TGFß1 Treatment of Megakaryocytes</title><title>Cellular physiology and biochemistry</title><addtitle>Cell Physiol Biochem</addtitle><description>Background: TGFβ1, a decisive regulator of megakaryocyte maturation and platelet formation, has previously been shown to up-regulate both, store operated Ca 2+ entry (SOCE) and Ca 2+ extrusion by Na + /Ca 2+ exchange. The growth factor thus augments the increase of cytosolic Ca 2+ activity ([Ca 2+ ] i ) following release of Ca 2+ from intracellular stores and accelerates the subsequent decline of [Ca 2+ ] i . The effect on SOCE is dependent on a signaling cascade including p38 kinase, serum & glucocorticoid inducible kinase SGK1, and nuclear factor NFκB. The specific Na + /Ca 2+ exchanger isoforms involved and the signalling regulating the Na + /Ca 2+ exchangers remained, however elusive. The present study explored, whether TGFβ1 influences the expression and function of K + insensitive (NCX) and K + sensitive (NCKX) Na + /Ca 2+ exchangers, and aimed to shed light on the signalling involved. Methods: In human megakaryocytic cells (MEG01) RT-PCR was performed to quantify NCX/NCKX isoform transcript levels, [Ca 2+ ] i was determined by Fura-2 fluorescence, and Na + /Ca 2+ exchanger activity was estimated from the increase of [Ca 2+ ] i following switch from an extracellular solution with 130 or 90 mM Na + and 0 mM Ca 2+ to an extracellular solution with 0 Na + and 2 mM Ca 2+ . K + concentration was 0 mM for analysis of NCX and 40 mM for analysis of NCKX. Results: TGFβ1 (60 ng/ml, 24 h) significantly increased the transcript levels of NCX1, NCKX1, NCKX2 and NCKX5. Moreover, TGFβ1 (60 ng/ml, 24 h) significantly increased the activity of both, NCX and NCKX. The effect of TGFβ1 on NCX and NCKX transcript levels and activity was significantly blunted by p38 kinase inhibitor Skepinone-L (1 µM), the effect on NCX and NCKX activity further by SGK1 inhibitor GSK-650394 (10 µM) and NFκB inhibitor Wogonin (100 µM). Conclusions: TGFβ1 markedly up-regulates transcription of NCX1, NCKX1, NCKX2, and NCKX5 and thus Na + /Ca 2+ exchanger activity, an effect requiring p38 kinase, SGK1 and NFκB.</description><subject>Benzoates - pharmacology</subject><subject>Bridged Bicyclo Compounds, Heterocyclic - pharmacology</subject><subject>Calcium</subject><subject>Calcium - metabolism</subject><subject>Cell Line</subject><subject>Dibenzocycloheptenes - pharmacology</subject><subject>Flavanones - pharmacology</subject><subject>Humans</subject><subject>Immediate-Early Proteins - antagonists & inhibitors</subject><subject>Immediate-Early Proteins - genetics</subject><subject>Immediate-Early Proteins - metabolism</subject><subject>Megakaryocytes - cytology</subject><subject>Megakaryocytes - drug effects</subject><subject>Megakaryocytes - metabolism</subject><subject>Microscopy, Fluorescence</subject><subject>NCKX1</subject><subject>NCKX2</subject><subject>NCKX5</subject><subject>NCX1</subject><subject>NF-kappa B - antagonists & inhibitors</subject><subject>NF-kappa B - metabolism</subject><subject>NF-κB</subject><subject>Original Paper</subject><subject>p38 kinase</subject><subject>p38 Mitogen-Activated Protein Kinases - antagonists & inhibitors</subject><subject>p38 Mitogen-Activated Protein Kinases - metabolism</subject><subject>Protein Isoforms - genetics</subject><subject>Protein Isoforms - metabolism</subject><subject>Protein-Serine-Threonine Kinases - antagonists & inhibitors</subject><subject>Protein-Serine-Threonine Kinases - genetics</subject><subject>Protein-Serine-Threonine Kinases - metabolism</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>SGK1</subject><subject>Sodium-Calcium Exchanger - genetics</subject><subject>Sodium-Calcium Exchanger - metabolism</subject><subject>Transcription, Genetic - drug effects</subject><subject>Transforming Growth Factor beta1 - pharmacology</subject><subject>Up-Regulation - drug effects</subject><issn>1015-8987</issn><issn>1421-9778</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>M--</sourceid><sourceid>EIF</sourceid><sourceid>DOA</sourceid><recordid>eNptkctu1DAUhi0EoqVlwR4hS6xQCfVxLraX7dAZql6oyrCOHPskpGTiyE6h8xJ9Bd6DLXueCQ8ps2Ll46NP35H-n5AXwN4B5OqQMZYJpRR_RHYh45AoIeTjODPIE6mk2CHPQrhh8SsUf0p2uJSQCpbtkvurVNKzttcB39JPizOgurf0cp78_nlM3-OAvcV-pJ-H5Bqb206Preupq-mlPjicaX5AT-7MF9036OM0eAxhA2wcR2Zsv7Xjms5d17nvbd_Q5WL-6wfQpUc9rjbaKLrARn_Vfu3MesSwT57Uugv4_OHdI9fzk-XsQ3L-cXE6OzpPDJeMJ5xzYaUALHgNaZaLXBc257lKK8kkQKFSLEBJmVW8qBXPuGDSIIAVxqZ75HSSWqdvysG3q3i_dLot_y6cb0rtx9Z0WGqMgSGktspFlma8qpgqOCAWaGSFRXS9mVzGuxA81lsfsHJTTrktJ7KvJna4rVZot-S_NiLwegJiIDHRLTC7Op4U5WDrSL38L_Vw5Q9ZspzP</recordid><startdate>2017</startdate><enddate>2017</enddate><creator>Al-Maghout, Tamer</creator><creator>Pelzl, Lisann</creator><creator>Sahu, Itishri</creator><creator>Sukkar, Basma</creator><creator>Hosseinzadeh, Zohreh</creator><creator>Gutti, Ravi</creator><creator>Laufer, Stefan</creator><creator>Voelkl, Jakob</creator><creator>Pieske, Burkert</creator><creator>Gawaz, Meinrad</creator><creator>Lang, Florian</creator><general>Cell Physiol Biochem Press GmbH & Co KG</general><scope>M--</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>DOA</scope></search><sort><creationdate>2017</creationdate><title>P38 Kinase, SGK1 and NF-κB Dependent Up-Regulation of Na+/Ca2+ Exchanger Expression and Activity Following TGFß1 Treatment of Megakaryocytes</title><author>Al-Maghout, Tamer ; Pelzl, Lisann ; Sahu, Itishri ; Sukkar, Basma ; Hosseinzadeh, Zohreh ; Gutti, Ravi ; Laufer, Stefan ; Voelkl, Jakob ; Pieske, Burkert ; Gawaz, Meinrad ; Lang, Florian</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c2802-2227d871e62f134575a6d52593b80811693e619884b26f9242708ce11d7cd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Benzoates - pharmacology</topic><topic>Bridged Bicyclo Compounds, Heterocyclic - pharmacology</topic><topic>Calcium</topic><topic>Calcium - metabolism</topic><topic>Cell Line</topic><topic>Dibenzocycloheptenes - pharmacology</topic><topic>Flavanones - pharmacology</topic><topic>Humans</topic><topic>Immediate-Early Proteins - antagonists & inhibitors</topic><topic>Immediate-Early Proteins - genetics</topic><topic>Immediate-Early Proteins - metabolism</topic><topic>Megakaryocytes - cytology</topic><topic>Megakaryocytes - drug effects</topic><topic>Megakaryocytes - metabolism</topic><topic>Microscopy, Fluorescence</topic><topic>NCKX1</topic><topic>NCKX2</topic><topic>NCKX5</topic><topic>NCX1</topic><topic>NF-kappa B - antagonists & inhibitors</topic><topic>NF-kappa B - metabolism</topic><topic>NF-κB</topic><topic>Original Paper</topic><topic>p38 kinase</topic><topic>p38 Mitogen-Activated Protein Kinases - antagonists & inhibitors</topic><topic>p38 Mitogen-Activated Protein Kinases - metabolism</topic><topic>Protein Isoforms - genetics</topic><topic>Protein Isoforms - metabolism</topic><topic>Protein-Serine-Threonine Kinases - antagonists & inhibitors</topic><topic>Protein-Serine-Threonine Kinases - genetics</topic><topic>Protein-Serine-Threonine Kinases - metabolism</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>SGK1</topic><topic>Sodium-Calcium Exchanger - genetics</topic><topic>Sodium-Calcium Exchanger - metabolism</topic><topic>Transcription, Genetic - drug effects</topic><topic>Transforming Growth Factor beta1 - pharmacology</topic><topic>Up-Regulation - drug effects</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Al-Maghout, Tamer</creatorcontrib><creatorcontrib>Pelzl, Lisann</creatorcontrib><creatorcontrib>Sahu, Itishri</creatorcontrib><creatorcontrib>Sukkar, Basma</creatorcontrib><creatorcontrib>Hosseinzadeh, Zohreh</creatorcontrib><creatorcontrib>Gutti, Ravi</creatorcontrib><creatorcontrib>Laufer, Stefan</creatorcontrib><creatorcontrib>Voelkl, Jakob</creatorcontrib><creatorcontrib>Pieske, Burkert</creatorcontrib><creatorcontrib>Gawaz, Meinrad</creatorcontrib><creatorcontrib>Lang, Florian</creatorcontrib><collection>Karger Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>Cellular physiology and biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Al-Maghout, Tamer</au><au>Pelzl, Lisann</au><au>Sahu, Itishri</au><au>Sukkar, Basma</au><au>Hosseinzadeh, Zohreh</au><au>Gutti, Ravi</au><au>Laufer, Stefan</au><au>Voelkl, Jakob</au><au>Pieske, Burkert</au><au>Gawaz, Meinrad</au><au>Lang, Florian</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>P38 Kinase, SGK1 and NF-κB Dependent Up-Regulation of Na+/Ca2+ Exchanger Expression and Activity Following TGFß1 Treatment of Megakaryocytes</atitle><jtitle>Cellular physiology and biochemistry</jtitle><addtitle>Cell Physiol Biochem</addtitle><date>2017</date><risdate>2017</risdate><volume>42</volume><issue>6</issue><spage>2169</spage><epage>2181</epage><pages>2169-2181</pages><issn>1015-8987</issn><eissn>1421-9778</eissn><abstract>Background: TGFβ1, a decisive regulator of megakaryocyte maturation and platelet formation, has previously been shown to up-regulate both, store operated Ca 2+ entry (SOCE) and Ca 2+ extrusion by Na + /Ca 2+ exchange. The growth factor thus augments the increase of cytosolic Ca 2+ activity ([Ca 2+ ] i ) following release of Ca 2+ from intracellular stores and accelerates the subsequent decline of [Ca 2+ ] i . The effect on SOCE is dependent on a signaling cascade including p38 kinase, serum & glucocorticoid inducible kinase SGK1, and nuclear factor NFκB. The specific Na + /Ca 2+ exchanger isoforms involved and the signalling regulating the Na + /Ca 2+ exchangers remained, however elusive. The present study explored, whether TGFβ1 influences the expression and function of K + insensitive (NCX) and K + sensitive (NCKX) Na + /Ca 2+ exchangers, and aimed to shed light on the signalling involved. Methods: In human megakaryocytic cells (MEG01) RT-PCR was performed to quantify NCX/NCKX isoform transcript levels, [Ca 2+ ] i was determined by Fura-2 fluorescence, and Na + /Ca 2+ exchanger activity was estimated from the increase of [Ca 2+ ] i following switch from an extracellular solution with 130 or 90 mM Na + and 0 mM Ca 2+ to an extracellular solution with 0 Na + and 2 mM Ca 2+ . K + concentration was 0 mM for analysis of NCX and 40 mM for analysis of NCKX. Results: TGFβ1 (60 ng/ml, 24 h) significantly increased the transcript levels of NCX1, NCKX1, NCKX2 and NCKX5. Moreover, TGFβ1 (60 ng/ml, 24 h) significantly increased the activity of both, NCX and NCKX. The effect of TGFβ1 on NCX and NCKX transcript levels and activity was significantly blunted by p38 kinase inhibitor Skepinone-L (1 µM), the effect on NCX and NCKX activity further by SGK1 inhibitor GSK-650394 (10 µM) and NFκB inhibitor Wogonin (100 µM). Conclusions: TGFβ1 markedly up-regulates transcription of NCX1, NCKX1, NCKX2, and NCKX5 and thus Na + /Ca 2+ exchanger activity, an effect requiring p38 kinase, SGK1 and NFκB.</abstract><cop>Basel, Switzerland</cop><pub>Cell Physiol Biochem Press GmbH & Co KG</pub><pmid>28813704</pmid><doi>10.1159/000479992</doi><tpages>13</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Benzoates - pharmacology Bridged Bicyclo Compounds, Heterocyclic - pharmacology Calcium Calcium - metabolism Cell Line Dibenzocycloheptenes - pharmacology Flavanones - pharmacology Humans Immediate-Early Proteins - antagonists & inhibitors Immediate-Early Proteins - genetics Immediate-Early Proteins - metabolism Megakaryocytes - cytology Megakaryocytes - drug effects Megakaryocytes - metabolism Microscopy, Fluorescence NCKX1 NCKX2 NCKX5 NCX1 NF-kappa B - antagonists & inhibitors NF-kappa B - metabolism NF-κB Original Paper p38 kinase p38 Mitogen-Activated Protein Kinases - antagonists & inhibitors p38 Mitogen-Activated Protein Kinases - metabolism Protein Isoforms - genetics Protein Isoforms - metabolism Protein-Serine-Threonine Kinases - antagonists & inhibitors Protein-Serine-Threonine Kinases - genetics Protein-Serine-Threonine Kinases - metabolism Real-Time Polymerase Chain Reaction SGK1 Sodium-Calcium Exchanger - genetics Sodium-Calcium Exchanger - metabolism Transcription, Genetic - drug effects Transforming Growth Factor beta1 - pharmacology Up-Regulation - drug effects |
| title | P38 Kinase, SGK1 and NF-κB Dependent Up-Regulation of Na+/Ca2+ Exchanger Expression and Activity Following TGFß1 Treatment of Megakaryocytes |
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