Loading…

High-Throughput Screening Identifies Genes Required for Candida albicans Induction of Macrophage Pyroptosis

The innate immune system is the first line of defense against invasive fungal infections. As a consequence, many successful fungal pathogens have evolved elegant strategies to interact with host immune cells. For example, undergoes a morphogenetic switch coupled to cell wall remodeling upon phagocyt...

Full description

Saved in:
Bibliographic Details
Published in:mBio 2018-08, Vol.9 (4)
Main Authors: O'Meara, Teresa R, Duah, Kwamaa, Guo, Cynthia X, Maxson, Michelle E, Gaudet, Ryan G, Koselny, Kristy, Wellington, Melanie, Powers, Michael E, MacAlpine, Jessie, O'Meara, Matthew J, Veri, Amanda O, Grinstein, Sergio, Noble, Suzanne M, Krysan, Damian, Gray-Owen, Scott D, Cowen, Leah E
Format: Article
Language:English
Subjects:
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
cited_by cdi_FETCH-LOGICAL-c387t-3aa5984540538b88ab1ad0b31122aca06e56845684c28424a2ff5d2944ad5d733
cites cdi_FETCH-LOGICAL-c387t-3aa5984540538b88ab1ad0b31122aca06e56845684c28424a2ff5d2944ad5d733
container_end_page
container_issue 4
container_start_page
container_title mBio
container_volume 9
creator O'Meara, Teresa R
Duah, Kwamaa
Guo, Cynthia X
Maxson, Michelle E
Gaudet, Ryan G
Koselny, Kristy
Wellington, Melanie
Powers, Michael E
MacAlpine, Jessie
O'Meara, Matthew J
Veri, Amanda O
Grinstein, Sergio
Noble, Suzanne M
Krysan, Damian
Gray-Owen, Scott D
Cowen, Leah E
description The innate immune system is the first line of defense against invasive fungal infections. As a consequence, many successful fungal pathogens have evolved elegant strategies to interact with host immune cells. For example, undergoes a morphogenetic switch coupled to cell wall remodeling upon phagocytosis by macrophages and then induces macrophage pyroptosis, an inflammatory cell death program. To elucidate the genetic circuitry through which orchestrates this host response, we performed the first large-scale analysis of interactions with mammalian immune cells. We identified 98 genes that enable macrophage pyroptosis without influencing fungal cell morphology in the macrophage, including specific determinants of cell wall biogenesis and the Hog1 signaling cascade. Using these mutated genes, we discovered that defects in the activation of pyroptosis affect immune cell recruitment during infection. Examining host circuitry required for pyroptosis in response to infection, we discovered that inflammasome priming and activation can be decoupled. Finally, we observed that poptosis-associated peck-like protein containing a ARD (ASC) oligomerization can occur prior to phagolysosomal rupture by hyphae, demonstrating that phagolysosomal rupture is not the inflammasome activating signal. Taking the data together, this work defines genes that enable fungal cell wall remodeling and activation of macrophage pyroptosis independently of effects on morphogenesis and identifies macrophage signaling components that are required for pyroptosis in response to infection. is a natural member of the human mucosal microbiota that can also cause superficial infections and life-threatening systemic infections, both of which are characterized by inflammation. Host defense relies mainly on the ingestion and destruction of by innate immune cells, such as macrophages and neutrophils. Although some cells are killed by macrophages, most undergo a morphological change and escape by inducing macrophage pyroptosis. Here, we investigated the genes and host factors that promote macrophage pyroptosis in response to intracellular fungi. This work provides a foundation for understanding how host immune cells interact with and may lead to effective strategies to modulate inflammation induced by fungal infections.
doi_str_mv 10.1128/mBio.01581-18
format article
fullrecord <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_6106084</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2091816764</sourcerecordid><originalsourceid>FETCH-LOGICAL-c387t-3aa5984540538b88ab1ad0b31122aca06e56845684c28424a2ff5d2944ad5d733</originalsourceid><addsrcrecordid>eNpVUU1r3DAQFSUlCdsccy069uJUI1le7SWQLE2ykNDSpmcxlmRbrVfaSHYh_z7a5oN2QKMH83ijp0fIKbAzAK4-by99PGMgFVSg3pFjDpJVSwlwsMcNVBz46oic5PyLlRIClGCH5EgwECAacUx-3_h-qO6HFOd-2M0T_WGSc8GHnm6sC5PvvMv02oXSv7uH2SdnaRcTXWOw3iLFsfUGQ6abYGcz-Rho7OgdmhR3A_aOfnssaIrZ5w_kfYdjdicv94L8vPpyv76pbr9eb9YXt5URajlVAlGuVC1rJoVqlcIW0LJWFMccDbLGyaaMyzFc1bxG3nXS8lVdo5V2KcSCnD_r7uZ266wpNhKOepf8FtOjjuj1_5PgB93HP7oB1jBVF4FPLwIpPswuT3rrs3HjiMHFOWvOVqCgWTZ7avVMLX5zTq57WwNM7zPS-4z034x0-f0F-fjv297Yr4mIJ1jXjnk</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2091816764</pqid></control><display><type>article</type><title>High-Throughput Screening Identifies Genes Required for Candida albicans Induction of Macrophage Pyroptosis</title><source>American Society for Microbiology Journals</source><source>PubMed Central</source><creator>O'Meara, Teresa R ; Duah, Kwamaa ; Guo, Cynthia X ; Maxson, Michelle E ; Gaudet, Ryan G ; Koselny, Kristy ; Wellington, Melanie ; Powers, Michael E ; MacAlpine, Jessie ; O'Meara, Matthew J ; Veri, Amanda O ; Grinstein, Sergio ; Noble, Suzanne M ; Krysan, Damian ; Gray-Owen, Scott D ; Cowen, Leah E</creator><contributor>Kronstad, James W.</contributor><creatorcontrib>O'Meara, Teresa R ; Duah, Kwamaa ; Guo, Cynthia X ; Maxson, Michelle E ; Gaudet, Ryan G ; Koselny, Kristy ; Wellington, Melanie ; Powers, Michael E ; MacAlpine, Jessie ; O'Meara, Matthew J ; Veri, Amanda O ; Grinstein, Sergio ; Noble, Suzanne M ; Krysan, Damian ; Gray-Owen, Scott D ; Cowen, Leah E ; Kronstad, James W.</creatorcontrib><description>The innate immune system is the first line of defense against invasive fungal infections. As a consequence, many successful fungal pathogens have evolved elegant strategies to interact with host immune cells. For example, undergoes a morphogenetic switch coupled to cell wall remodeling upon phagocytosis by macrophages and then induces macrophage pyroptosis, an inflammatory cell death program. To elucidate the genetic circuitry through which orchestrates this host response, we performed the first large-scale analysis of interactions with mammalian immune cells. We identified 98 genes that enable macrophage pyroptosis without influencing fungal cell morphology in the macrophage, including specific determinants of cell wall biogenesis and the Hog1 signaling cascade. Using these mutated genes, we discovered that defects in the activation of pyroptosis affect immune cell recruitment during infection. Examining host circuitry required for pyroptosis in response to infection, we discovered that inflammasome priming and activation can be decoupled. Finally, we observed that poptosis-associated peck-like protein containing a ARD (ASC) oligomerization can occur prior to phagolysosomal rupture by hyphae, demonstrating that phagolysosomal rupture is not the inflammasome activating signal. Taking the data together, this work defines genes that enable fungal cell wall remodeling and activation of macrophage pyroptosis independently of effects on morphogenesis and identifies macrophage signaling components that are required for pyroptosis in response to infection. is a natural member of the human mucosal microbiota that can also cause superficial infections and life-threatening systemic infections, both of which are characterized by inflammation. Host defense relies mainly on the ingestion and destruction of by innate immune cells, such as macrophages and neutrophils. Although some cells are killed by macrophages, most undergo a morphological change and escape by inducing macrophage pyroptosis. Here, we investigated the genes and host factors that promote macrophage pyroptosis in response to intracellular fungi. This work provides a foundation for understanding how host immune cells interact with and may lead to effective strategies to modulate inflammation induced by fungal infections.</description><identifier>ISSN: 2161-2129</identifier><identifier>EISSN: 2150-7511</identifier><identifier>DOI: 10.1128/mBio.01581-18</identifier><identifier>PMID: 30131363</identifier><language>eng</language><publisher>United States: American Society for Microbiology</publisher><subject>Animals ; Candida albicans - genetics ; Candida albicans - pathogenicity ; Female ; Genes, Fungal ; High-Throughput Screening Assays ; Host-Pathogen Interactions ; Immune Evasion ; Macrophages - microbiology ; Macrophages - pathology ; Mice ; Mice, Inbred C57BL ; Phagocytosis ; Pyroptosis</subject><ispartof>mBio, 2018-08, Vol.9 (4)</ispartof><rights>Copyright © 2018 O’Meara et al.</rights><rights>Copyright © 2018 O’Meara et al. 2018 O’Meara et al.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c387t-3aa5984540538b88ab1ad0b31122aca06e56845684c28424a2ff5d2944ad5d733</citedby><cites>FETCH-LOGICAL-c387t-3aa5984540538b88ab1ad0b31122aca06e56845684c28424a2ff5d2944ad5d733</cites><orcidid>0000-0002-1477-3616</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6106084/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6106084/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,3188,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30131363$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Kronstad, James W.</contributor><creatorcontrib>O'Meara, Teresa R</creatorcontrib><creatorcontrib>Duah, Kwamaa</creatorcontrib><creatorcontrib>Guo, Cynthia X</creatorcontrib><creatorcontrib>Maxson, Michelle E</creatorcontrib><creatorcontrib>Gaudet, Ryan G</creatorcontrib><creatorcontrib>Koselny, Kristy</creatorcontrib><creatorcontrib>Wellington, Melanie</creatorcontrib><creatorcontrib>Powers, Michael E</creatorcontrib><creatorcontrib>MacAlpine, Jessie</creatorcontrib><creatorcontrib>O'Meara, Matthew J</creatorcontrib><creatorcontrib>Veri, Amanda O</creatorcontrib><creatorcontrib>Grinstein, Sergio</creatorcontrib><creatorcontrib>Noble, Suzanne M</creatorcontrib><creatorcontrib>Krysan, Damian</creatorcontrib><creatorcontrib>Gray-Owen, Scott D</creatorcontrib><creatorcontrib>Cowen, Leah E</creatorcontrib><title>High-Throughput Screening Identifies Genes Required for Candida albicans Induction of Macrophage Pyroptosis</title><title>mBio</title><addtitle>mBio</addtitle><description>The innate immune system is the first line of defense against invasive fungal infections. As a consequence, many successful fungal pathogens have evolved elegant strategies to interact with host immune cells. For example, undergoes a morphogenetic switch coupled to cell wall remodeling upon phagocytosis by macrophages and then induces macrophage pyroptosis, an inflammatory cell death program. To elucidate the genetic circuitry through which orchestrates this host response, we performed the first large-scale analysis of interactions with mammalian immune cells. We identified 98 genes that enable macrophage pyroptosis without influencing fungal cell morphology in the macrophage, including specific determinants of cell wall biogenesis and the Hog1 signaling cascade. Using these mutated genes, we discovered that defects in the activation of pyroptosis affect immune cell recruitment during infection. Examining host circuitry required for pyroptosis in response to infection, we discovered that inflammasome priming and activation can be decoupled. Finally, we observed that poptosis-associated peck-like protein containing a ARD (ASC) oligomerization can occur prior to phagolysosomal rupture by hyphae, demonstrating that phagolysosomal rupture is not the inflammasome activating signal. Taking the data together, this work defines genes that enable fungal cell wall remodeling and activation of macrophage pyroptosis independently of effects on morphogenesis and identifies macrophage signaling components that are required for pyroptosis in response to infection. is a natural member of the human mucosal microbiota that can also cause superficial infections and life-threatening systemic infections, both of which are characterized by inflammation. Host defense relies mainly on the ingestion and destruction of by innate immune cells, such as macrophages and neutrophils. Although some cells are killed by macrophages, most undergo a morphological change and escape by inducing macrophage pyroptosis. Here, we investigated the genes and host factors that promote macrophage pyroptosis in response to intracellular fungi. This work provides a foundation for understanding how host immune cells interact with and may lead to effective strategies to modulate inflammation induced by fungal infections.</description><subject>Animals</subject><subject>Candida albicans - genetics</subject><subject>Candida albicans - pathogenicity</subject><subject>Female</subject><subject>Genes, Fungal</subject><subject>High-Throughput Screening Assays</subject><subject>Host-Pathogen Interactions</subject><subject>Immune Evasion</subject><subject>Macrophages - microbiology</subject><subject>Macrophages - pathology</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Phagocytosis</subject><subject>Pyroptosis</subject><issn>2161-2129</issn><issn>2150-7511</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><recordid>eNpVUU1r3DAQFSUlCdsccy069uJUI1le7SWQLE2ykNDSpmcxlmRbrVfaSHYh_z7a5oN2QKMH83ijp0fIKbAzAK4-by99PGMgFVSg3pFjDpJVSwlwsMcNVBz46oic5PyLlRIClGCH5EgwECAacUx-3_h-qO6HFOd-2M0T_WGSc8GHnm6sC5PvvMv02oXSv7uH2SdnaRcTXWOw3iLFsfUGQ6abYGcz-Rho7OgdmhR3A_aOfnssaIrZ5w_kfYdjdicv94L8vPpyv76pbr9eb9YXt5URajlVAlGuVC1rJoVqlcIW0LJWFMccDbLGyaaMyzFc1bxG3nXS8lVdo5V2KcSCnD_r7uZ266wpNhKOepf8FtOjjuj1_5PgB93HP7oB1jBVF4FPLwIpPswuT3rrs3HjiMHFOWvOVqCgWTZ7avVMLX5zTq57WwNM7zPS-4z034x0-f0F-fjv297Yr4mIJ1jXjnk</recordid><startdate>20180821</startdate><enddate>20180821</enddate><creator>O'Meara, Teresa R</creator><creator>Duah, Kwamaa</creator><creator>Guo, Cynthia X</creator><creator>Maxson, Michelle E</creator><creator>Gaudet, Ryan G</creator><creator>Koselny, Kristy</creator><creator>Wellington, Melanie</creator><creator>Powers, Michael E</creator><creator>MacAlpine, Jessie</creator><creator>O'Meara, Matthew J</creator><creator>Veri, Amanda O</creator><creator>Grinstein, Sergio</creator><creator>Noble, Suzanne M</creator><creator>Krysan, Damian</creator><creator>Gray-Owen, Scott D</creator><creator>Cowen, Leah E</creator><general>American Society for Microbiology</general><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>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0002-1477-3616</orcidid></search><sort><creationdate>20180821</creationdate><title>High-Throughput Screening Identifies Genes Required for Candida albicans Induction of Macrophage Pyroptosis</title><author>O'Meara, Teresa R ; Duah, Kwamaa ; Guo, Cynthia X ; Maxson, Michelle E ; Gaudet, Ryan G ; Koselny, Kristy ; Wellington, Melanie ; Powers, Michael E ; MacAlpine, Jessie ; O'Meara, Matthew J ; Veri, Amanda O ; Grinstein, Sergio ; Noble, Suzanne M ; Krysan, Damian ; Gray-Owen, Scott D ; Cowen, Leah E</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c387t-3aa5984540538b88ab1ad0b31122aca06e56845684c28424a2ff5d2944ad5d733</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Animals</topic><topic>Candida albicans - genetics</topic><topic>Candida albicans - pathogenicity</topic><topic>Female</topic><topic>Genes, Fungal</topic><topic>High-Throughput Screening Assays</topic><topic>Host-Pathogen Interactions</topic><topic>Immune Evasion</topic><topic>Macrophages - microbiology</topic><topic>Macrophages - pathology</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Phagocytosis</topic><topic>Pyroptosis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>O'Meara, Teresa R</creatorcontrib><creatorcontrib>Duah, Kwamaa</creatorcontrib><creatorcontrib>Guo, Cynthia X</creatorcontrib><creatorcontrib>Maxson, Michelle E</creatorcontrib><creatorcontrib>Gaudet, Ryan G</creatorcontrib><creatorcontrib>Koselny, Kristy</creatorcontrib><creatorcontrib>Wellington, Melanie</creatorcontrib><creatorcontrib>Powers, Michael E</creatorcontrib><creatorcontrib>MacAlpine, Jessie</creatorcontrib><creatorcontrib>O'Meara, Matthew J</creatorcontrib><creatorcontrib>Veri, Amanda O</creatorcontrib><creatorcontrib>Grinstein, Sergio</creatorcontrib><creatorcontrib>Noble, Suzanne M</creatorcontrib><creatorcontrib>Krysan, Damian</creatorcontrib><creatorcontrib>Gray-Owen, Scott D</creatorcontrib><creatorcontrib>Cowen, Leah E</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>mBio</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>O'Meara, Teresa R</au><au>Duah, Kwamaa</au><au>Guo, Cynthia X</au><au>Maxson, Michelle E</au><au>Gaudet, Ryan G</au><au>Koselny, Kristy</au><au>Wellington, Melanie</au><au>Powers, Michael E</au><au>MacAlpine, Jessie</au><au>O'Meara, Matthew J</au><au>Veri, Amanda O</au><au>Grinstein, Sergio</au><au>Noble, Suzanne M</au><au>Krysan, Damian</au><au>Gray-Owen, Scott D</au><au>Cowen, Leah E</au><au>Kronstad, James W.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>High-Throughput Screening Identifies Genes Required for Candida albicans Induction of Macrophage Pyroptosis</atitle><jtitle>mBio</jtitle><addtitle>mBio</addtitle><date>2018-08-21</date><risdate>2018</risdate><volume>9</volume><issue>4</issue><issn>2161-2129</issn><eissn>2150-7511</eissn><abstract>The innate immune system is the first line of defense against invasive fungal infections. As a consequence, many successful fungal pathogens have evolved elegant strategies to interact with host immune cells. For example, undergoes a morphogenetic switch coupled to cell wall remodeling upon phagocytosis by macrophages and then induces macrophage pyroptosis, an inflammatory cell death program. To elucidate the genetic circuitry through which orchestrates this host response, we performed the first large-scale analysis of interactions with mammalian immune cells. We identified 98 genes that enable macrophage pyroptosis without influencing fungal cell morphology in the macrophage, including specific determinants of cell wall biogenesis and the Hog1 signaling cascade. Using these mutated genes, we discovered that defects in the activation of pyroptosis affect immune cell recruitment during infection. Examining host circuitry required for pyroptosis in response to infection, we discovered that inflammasome priming and activation can be decoupled. Finally, we observed that poptosis-associated peck-like protein containing a ARD (ASC) oligomerization can occur prior to phagolysosomal rupture by hyphae, demonstrating that phagolysosomal rupture is not the inflammasome activating signal. Taking the data together, this work defines genes that enable fungal cell wall remodeling and activation of macrophage pyroptosis independently of effects on morphogenesis and identifies macrophage signaling components that are required for pyroptosis in response to infection. is a natural member of the human mucosal microbiota that can also cause superficial infections and life-threatening systemic infections, both of which are characterized by inflammation. Host defense relies mainly on the ingestion and destruction of by innate immune cells, such as macrophages and neutrophils. Although some cells are killed by macrophages, most undergo a morphological change and escape by inducing macrophage pyroptosis. Here, we investigated the genes and host factors that promote macrophage pyroptosis in response to intracellular fungi. This work provides a foundation for understanding how host immune cells interact with and may lead to effective strategies to modulate inflammation induced by fungal infections.</abstract><cop>United States</cop><pub>American Society for Microbiology</pub><pmid>30131363</pmid><doi>10.1128/mBio.01581-18</doi><orcidid>https://orcid.org/0000-0002-1477-3616</orcidid><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 2161-2129
ispartof mBio, 2018-08, Vol.9 (4)
issn 2161-2129
2150-7511
language eng
recordid cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_6106084
source American Society for Microbiology Journals; PubMed Central
subjects Animals
Candida albicans - genetics
Candida albicans - pathogenicity
Female
Genes, Fungal
High-Throughput Screening Assays
Host-Pathogen Interactions
Immune Evasion
Macrophages - microbiology
Macrophages - pathology
Mice
Mice, Inbred C57BL
Phagocytosis
Pyroptosis
title High-Throughput Screening Identifies Genes Required for Candida albicans Induction of Macrophage Pyroptosis
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T11%3A58%3A19IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=High-Throughput%20Screening%20Identifies%20Genes%20Required%20for%20Candida%20albicans%20Induction%20of%20Macrophage%20Pyroptosis&rft.jtitle=mBio&rft.au=O'Meara,%20Teresa%20R&rft.date=2018-08-21&rft.volume=9&rft.issue=4&rft.issn=2161-2129&rft.eissn=2150-7511&rft_id=info:doi/10.1128/mBio.01581-18&rft_dat=%3Cproquest_pubme%3E2091816764%3C/proquest_pubme%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c387t-3aa5984540538b88ab1ad0b31122aca06e56845684c28424a2ff5d2944ad5d733%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=2091816764&rft_id=info:pmid/30131363&rfr_iscdi=true