Loading…
A zebrafish model of nondystrophic myotonia with sodium channelopathy
•Nondystrophic myotonias are disorders of Na+ and Cl− channels in skeletal muscles.•We generated transgenic zebrafish expressing a human mutant allele of Na+ channels.•The transgenic zebrafish showed electromyographic features of transgenic zebrafish.•The zebrafish displayed swimming behavior sugges...
Saved in:
| Published in: | Neuroscience letters 2020-01, Vol.714, p.134579-134579, Article 134579 |
|---|---|
| Main Authors: | , , , , , , , , , , , , , |
| 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-c362t-45008bd16e94057684d00cac657d2581b576ca90bbb4d569a20320d08a2013c53 |
|---|---|
| cites | cdi_FETCH-LOGICAL-c362t-45008bd16e94057684d00cac657d2581b576ca90bbb4d569a20320d08a2013c53 |
| container_end_page | 134579 |
| container_issue | |
| container_start_page | 134579 |
| container_title | Neuroscience letters |
| container_volume | 714 |
| creator | Nam, Tai-Seung Zhang, Jun Chandrasekaran, Gopalakrishnan Jeong, In Young Li, Wenting Lee, So-Hyun Kang, Kyung-Wook Maeng, Jin-Soo Kang, Hyuno Shin, Hee-Young Park, Hae-Chul Kim, Sohee Choi, Seok-Yong Kim, Myeong-Kyu |
| description | •Nondystrophic myotonias are disorders of Na+ and Cl− channels in skeletal muscles.•We generated transgenic zebrafish expressing a human mutant allele of Na+ channels.•The transgenic zebrafish showed electromyographic features of transgenic zebrafish.•The zebrafish displayed swimming behavior suggestive of nondystrophic myotonias.•This zebrafish model is a good vertebrate model of human nondystrophic myotonias.
Nondystrophic myotonias are disorders of Na+ (Nav1.4 or SCN4A) and Cl− (CLCN1) channels in skeletal muscles, and frequently show phenotype heterogeneity. The molecular mechanism underlying their pathophysiology and phenotype heterogeneity remains unclear. As zebrafish models have been recently exploited for studies of the pathophysiology and phenotype heterogeneity of various human genetic diseases, a zebrafish model may be useful for delineating nondystrophic myotonias. Here, we generated transgenic zebrafish expressing a human mutant allele of SCN4A, referred to as Tg(mylpfa:N440K), and needle electromyography revealed increased number of myotonic discharges and positive sharp waves in the muscles of Tg(mylpfa:N440K) than in controls. In addition, forced exercise test at a water temperature of 24 °C showed a decrease in the distance moved, time spent in and number of visits to the zone with stronger swimming resistance. Finally, a forced exercise test at a water temperature of 18 °C exhibited a higher number of dive-bombing periods and drifting-down behavior than in controls. These findings indicate that Tg(mylpfa:N440K) is a good vertebrate model of exercise- and cold-induced human nondystrophic myotonias. This zebrafish model may contribute to provide insight into the pathophysiology of myotonia in sodium channelopathy and could be used to explore a new therapeutic avenue. |
| doi_str_mv | 10.1016/j.neulet.2019.134579 |
| format | article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2310721801</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0304394019306822</els_id><sourcerecordid>2310721801</sourcerecordid><originalsourceid>FETCH-LOGICAL-c362t-45008bd16e94057684d00cac657d2581b576ca90bbb4d569a20320d08a2013c53</originalsourceid><addsrcrecordid>eNp9kMtOwzAQRS0EoqXwBwhlySZlbMd5bJAqVB5SJTawthx7qrhK4mInoPD1pEphyWpGo3vnzhxCriksKdD0brdssa-xWzKgxZLyRGTFCZnTPGNxVmTslMyBQxLzIoEZuQhhBwCCiuSczDhN04JTMSfrVfSNpVdbG6qocQbryG2j1rVmCJ13-8rqqBlc51qroi_bVVFwxvZNpCvVtli7veqq4ZKcbVUd8OpYF-T9cf328BxvXp9eHlabWPOUdXEiAPLS0BTHm0SW5okB0EqnIjNM5LQcZ1oVUJZlYkRaKAacgYF8bCjXgi_I7bR3791Hj6GTjQ0a61q16PogGaeQMZqP6gVJJqn2LgSPW7n3tlF-kBTkAaDcyQmgPACUE8DRdnNM6MsGzZ_pl9gouJ8EOP75adHLoC22Go31qDtpnP0_4QehiYK9</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2310721801</pqid></control><display><type>article</type><title>A zebrafish model of nondystrophic myotonia with sodium channelopathy</title><source>ScienceDirect Freedom Collection</source><creator>Nam, Tai-Seung ; Zhang, Jun ; Chandrasekaran, Gopalakrishnan ; Jeong, In Young ; Li, Wenting ; Lee, So-Hyun ; Kang, Kyung-Wook ; Maeng, Jin-Soo ; Kang, Hyuno ; Shin, Hee-Young ; Park, Hae-Chul ; Kim, Sohee ; Choi, Seok-Yong ; Kim, Myeong-Kyu</creator><creatorcontrib>Nam, Tai-Seung ; Zhang, Jun ; Chandrasekaran, Gopalakrishnan ; Jeong, In Young ; Li, Wenting ; Lee, So-Hyun ; Kang, Kyung-Wook ; Maeng, Jin-Soo ; Kang, Hyuno ; Shin, Hee-Young ; Park, Hae-Chul ; Kim, Sohee ; Choi, Seok-Yong ; Kim, Myeong-Kyu</creatorcontrib><description>•Nondystrophic myotonias are disorders of Na+ and Cl− channels in skeletal muscles.•We generated transgenic zebrafish expressing a human mutant allele of Na+ channels.•The transgenic zebrafish showed electromyographic features of transgenic zebrafish.•The zebrafish displayed swimming behavior suggestive of nondystrophic myotonias.•This zebrafish model is a good vertebrate model of human nondystrophic myotonias.
Nondystrophic myotonias are disorders of Na+ (Nav1.4 or SCN4A) and Cl− (CLCN1) channels in skeletal muscles, and frequently show phenotype heterogeneity. The molecular mechanism underlying their pathophysiology and phenotype heterogeneity remains unclear. As zebrafish models have been recently exploited for studies of the pathophysiology and phenotype heterogeneity of various human genetic diseases, a zebrafish model may be useful for delineating nondystrophic myotonias. Here, we generated transgenic zebrafish expressing a human mutant allele of SCN4A, referred to as Tg(mylpfa:N440K), and needle electromyography revealed increased number of myotonic discharges and positive sharp waves in the muscles of Tg(mylpfa:N440K) than in controls. In addition, forced exercise test at a water temperature of 24 °C showed a decrease in the distance moved, time spent in and number of visits to the zone with stronger swimming resistance. Finally, a forced exercise test at a water temperature of 18 °C exhibited a higher number of dive-bombing periods and drifting-down behavior than in controls. These findings indicate that Tg(mylpfa:N440K) is a good vertebrate model of exercise- and cold-induced human nondystrophic myotonias. This zebrafish model may contribute to provide insight into the pathophysiology of myotonia in sodium channelopathy and could be used to explore a new therapeutic avenue.</description><identifier>ISSN: 0304-3940</identifier><identifier>EISSN: 1872-7972</identifier><identifier>DOI: 10.1016/j.neulet.2019.134579</identifier><identifier>PMID: 31669315</identifier><language>eng</language><publisher>Ireland: Elsevier B.V</publisher><subject>Animals ; Animals, Genetically Modified ; Cold Temperature ; Disease Models, Animal ; Electromyography ; Hyperkalemic periodic paralysis ; Muscle, Skeletal - physiopathology ; Mutation, Missense ; Myotonia ; Myotonia - genetics ; Myotonia - physiopathology ; Myotonia Congenita - genetics ; Myotonia Congenita - physiopathology ; Myotonic Disorders - genetics ; Myotonic Disorders - physiopathology ; NAV1.4 Voltage-Gated Sodium Channel - genetics ; Paralysis, Hyperkalemic Periodic - genetics ; Paralysis, Hyperkalemic Periodic - physiopathology ; Paramyotonia congenita ; Physical Exertion ; Sodium channel ; Zebrafish</subject><ispartof>Neuroscience letters, 2020-01, Vol.714, p.134579-134579, Article 134579</ispartof><rights>2019 Elsevier B.V.</rights><rights>Copyright © 2019 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c362t-45008bd16e94057684d00cac657d2581b576ca90bbb4d569a20320d08a2013c53</citedby><cites>FETCH-LOGICAL-c362t-45008bd16e94057684d00cac657d2581b576ca90bbb4d569a20320d08a2013c53</cites><orcidid>0000-0001-9032-9907 ; 0000-0001-7193-292X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,786,790,27957,27958</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31669315$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nam, Tai-Seung</creatorcontrib><creatorcontrib>Zhang, Jun</creatorcontrib><creatorcontrib>Chandrasekaran, Gopalakrishnan</creatorcontrib><creatorcontrib>Jeong, In Young</creatorcontrib><creatorcontrib>Li, Wenting</creatorcontrib><creatorcontrib>Lee, So-Hyun</creatorcontrib><creatorcontrib>Kang, Kyung-Wook</creatorcontrib><creatorcontrib>Maeng, Jin-Soo</creatorcontrib><creatorcontrib>Kang, Hyuno</creatorcontrib><creatorcontrib>Shin, Hee-Young</creatorcontrib><creatorcontrib>Park, Hae-Chul</creatorcontrib><creatorcontrib>Kim, Sohee</creatorcontrib><creatorcontrib>Choi, Seok-Yong</creatorcontrib><creatorcontrib>Kim, Myeong-Kyu</creatorcontrib><title>A zebrafish model of nondystrophic myotonia with sodium channelopathy</title><title>Neuroscience letters</title><addtitle>Neurosci Lett</addtitle><description>•Nondystrophic myotonias are disorders of Na+ and Cl− channels in skeletal muscles.•We generated transgenic zebrafish expressing a human mutant allele of Na+ channels.•The transgenic zebrafish showed electromyographic features of transgenic zebrafish.•The zebrafish displayed swimming behavior suggestive of nondystrophic myotonias.•This zebrafish model is a good vertebrate model of human nondystrophic myotonias.
Nondystrophic myotonias are disorders of Na+ (Nav1.4 or SCN4A) and Cl− (CLCN1) channels in skeletal muscles, and frequently show phenotype heterogeneity. The molecular mechanism underlying their pathophysiology and phenotype heterogeneity remains unclear. As zebrafish models have been recently exploited for studies of the pathophysiology and phenotype heterogeneity of various human genetic diseases, a zebrafish model may be useful for delineating nondystrophic myotonias. Here, we generated transgenic zebrafish expressing a human mutant allele of SCN4A, referred to as Tg(mylpfa:N440K), and needle electromyography revealed increased number of myotonic discharges and positive sharp waves in the muscles of Tg(mylpfa:N440K) than in controls. In addition, forced exercise test at a water temperature of 24 °C showed a decrease in the distance moved, time spent in and number of visits to the zone with stronger swimming resistance. Finally, a forced exercise test at a water temperature of 18 °C exhibited a higher number of dive-bombing periods and drifting-down behavior than in controls. These findings indicate that Tg(mylpfa:N440K) is a good vertebrate model of exercise- and cold-induced human nondystrophic myotonias. This zebrafish model may contribute to provide insight into the pathophysiology of myotonia in sodium channelopathy and could be used to explore a new therapeutic avenue.</description><subject>Animals</subject><subject>Animals, Genetically Modified</subject><subject>Cold Temperature</subject><subject>Disease Models, Animal</subject><subject>Electromyography</subject><subject>Hyperkalemic periodic paralysis</subject><subject>Muscle, Skeletal - physiopathology</subject><subject>Mutation, Missense</subject><subject>Myotonia</subject><subject>Myotonia - genetics</subject><subject>Myotonia - physiopathology</subject><subject>Myotonia Congenita - genetics</subject><subject>Myotonia Congenita - physiopathology</subject><subject>Myotonic Disorders - genetics</subject><subject>Myotonic Disorders - physiopathology</subject><subject>NAV1.4 Voltage-Gated Sodium Channel - genetics</subject><subject>Paralysis, Hyperkalemic Periodic - genetics</subject><subject>Paralysis, Hyperkalemic Periodic - physiopathology</subject><subject>Paramyotonia congenita</subject><subject>Physical Exertion</subject><subject>Sodium channel</subject><subject>Zebrafish</subject><issn>0304-3940</issn><issn>1872-7972</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNp9kMtOwzAQRS0EoqXwBwhlySZlbMd5bJAqVB5SJTawthx7qrhK4mInoPD1pEphyWpGo3vnzhxCriksKdD0brdssa-xWzKgxZLyRGTFCZnTPGNxVmTslMyBQxLzIoEZuQhhBwCCiuSczDhN04JTMSfrVfSNpVdbG6qocQbryG2j1rVmCJ13-8rqqBlc51qroi_bVVFwxvZNpCvVtli7veqq4ZKcbVUd8OpYF-T9cf328BxvXp9eHlabWPOUdXEiAPLS0BTHm0SW5okB0EqnIjNM5LQcZ1oVUJZlYkRaKAacgYF8bCjXgi_I7bR3791Hj6GTjQ0a61q16PogGaeQMZqP6gVJJqn2LgSPW7n3tlF-kBTkAaDcyQmgPACUE8DRdnNM6MsGzZ_pl9gouJ8EOP75adHLoC22Go31qDtpnP0_4QehiYK9</recordid><startdate>20200101</startdate><enddate>20200101</enddate><creator>Nam, Tai-Seung</creator><creator>Zhang, Jun</creator><creator>Chandrasekaran, Gopalakrishnan</creator><creator>Jeong, In Young</creator><creator>Li, Wenting</creator><creator>Lee, So-Hyun</creator><creator>Kang, Kyung-Wook</creator><creator>Maeng, Jin-Soo</creator><creator>Kang, Hyuno</creator><creator>Shin, Hee-Young</creator><creator>Park, Hae-Chul</creator><creator>Kim, Sohee</creator><creator>Choi, Seok-Yong</creator><creator>Kim, Myeong-Kyu</creator><general>Elsevier B.V</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><orcidid>https://orcid.org/0000-0001-9032-9907</orcidid><orcidid>https://orcid.org/0000-0001-7193-292X</orcidid></search><sort><creationdate>20200101</creationdate><title>A zebrafish model of nondystrophic myotonia with sodium channelopathy</title><author>Nam, Tai-Seung ; Zhang, Jun ; Chandrasekaran, Gopalakrishnan ; Jeong, In Young ; Li, Wenting ; Lee, So-Hyun ; Kang, Kyung-Wook ; Maeng, Jin-Soo ; Kang, Hyuno ; Shin, Hee-Young ; Park, Hae-Chul ; Kim, Sohee ; Choi, Seok-Yong ; Kim, Myeong-Kyu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c362t-45008bd16e94057684d00cac657d2581b576ca90bbb4d569a20320d08a2013c53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Animals</topic><topic>Animals, Genetically Modified</topic><topic>Cold Temperature</topic><topic>Disease Models, Animal</topic><topic>Electromyography</topic><topic>Hyperkalemic periodic paralysis</topic><topic>Muscle, Skeletal - physiopathology</topic><topic>Mutation, Missense</topic><topic>Myotonia</topic><topic>Myotonia - genetics</topic><topic>Myotonia - physiopathology</topic><topic>Myotonia Congenita - genetics</topic><topic>Myotonia Congenita - physiopathology</topic><topic>Myotonic Disorders - genetics</topic><topic>Myotonic Disorders - physiopathology</topic><topic>NAV1.4 Voltage-Gated Sodium Channel - genetics</topic><topic>Paralysis, Hyperkalemic Periodic - genetics</topic><topic>Paralysis, Hyperkalemic Periodic - physiopathology</topic><topic>Paramyotonia congenita</topic><topic>Physical Exertion</topic><topic>Sodium channel</topic><topic>Zebrafish</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nam, Tai-Seung</creatorcontrib><creatorcontrib>Zhang, Jun</creatorcontrib><creatorcontrib>Chandrasekaran, Gopalakrishnan</creatorcontrib><creatorcontrib>Jeong, In Young</creatorcontrib><creatorcontrib>Li, Wenting</creatorcontrib><creatorcontrib>Lee, So-Hyun</creatorcontrib><creatorcontrib>Kang, Kyung-Wook</creatorcontrib><creatorcontrib>Maeng, Jin-Soo</creatorcontrib><creatorcontrib>Kang, Hyuno</creatorcontrib><creatorcontrib>Shin, Hee-Young</creatorcontrib><creatorcontrib>Park, Hae-Chul</creatorcontrib><creatorcontrib>Kim, Sohee</creatorcontrib><creatorcontrib>Choi, Seok-Yong</creatorcontrib><creatorcontrib>Kim, Myeong-Kyu</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><jtitle>Neuroscience letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nam, Tai-Seung</au><au>Zhang, Jun</au><au>Chandrasekaran, Gopalakrishnan</au><au>Jeong, In Young</au><au>Li, Wenting</au><au>Lee, So-Hyun</au><au>Kang, Kyung-Wook</au><au>Maeng, Jin-Soo</au><au>Kang, Hyuno</au><au>Shin, Hee-Young</au><au>Park, Hae-Chul</au><au>Kim, Sohee</au><au>Choi, Seok-Yong</au><au>Kim, Myeong-Kyu</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A zebrafish model of nondystrophic myotonia with sodium channelopathy</atitle><jtitle>Neuroscience letters</jtitle><addtitle>Neurosci Lett</addtitle><date>2020-01-01</date><risdate>2020</risdate><volume>714</volume><spage>134579</spage><epage>134579</epage><pages>134579-134579</pages><artnum>134579</artnum><issn>0304-3940</issn><eissn>1872-7972</eissn><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><abstract>•Nondystrophic myotonias are disorders of Na+ and Cl− channels in skeletal muscles.•We generated transgenic zebrafish expressing a human mutant allele of Na+ channels.•The transgenic zebrafish showed electromyographic features of transgenic zebrafish.•The zebrafish displayed swimming behavior suggestive of nondystrophic myotonias.•This zebrafish model is a good vertebrate model of human nondystrophic myotonias.
Nondystrophic myotonias are disorders of Na+ (Nav1.4 or SCN4A) and Cl− (CLCN1) channels in skeletal muscles, and frequently show phenotype heterogeneity. The molecular mechanism underlying their pathophysiology and phenotype heterogeneity remains unclear. As zebrafish models have been recently exploited for studies of the pathophysiology and phenotype heterogeneity of various human genetic diseases, a zebrafish model may be useful for delineating nondystrophic myotonias. Here, we generated transgenic zebrafish expressing a human mutant allele of SCN4A, referred to as Tg(mylpfa:N440K), and needle electromyography revealed increased number of myotonic discharges and positive sharp waves in the muscles of Tg(mylpfa:N440K) than in controls. In addition, forced exercise test at a water temperature of 24 °C showed a decrease in the distance moved, time spent in and number of visits to the zone with stronger swimming resistance. Finally, a forced exercise test at a water temperature of 18 °C exhibited a higher number of dive-bombing periods and drifting-down behavior than in controls. These findings indicate that Tg(mylpfa:N440K) is a good vertebrate model of exercise- and cold-induced human nondystrophic myotonias. This zebrafish model may contribute to provide insight into the pathophysiology of myotonia in sodium channelopathy and could be used to explore a new therapeutic avenue.</abstract><cop>Ireland</cop><pub>Elsevier B.V</pub><pmid>31669315</pmid><doi>10.1016/j.neulet.2019.134579</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0001-9032-9907</orcidid><orcidid>https://orcid.org/0000-0001-7193-292X</orcidid></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0304-3940 |
| ispartof | Neuroscience letters, 2020-01, Vol.714, p.134579-134579, Article 134579 |
| issn | 0304-3940 1872-7972 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_2310721801 |
| source | ScienceDirect Freedom Collection |
| subjects | Animals Animals, Genetically Modified Cold Temperature Disease Models, Animal Electromyography Hyperkalemic periodic paralysis Muscle, Skeletal - physiopathology Mutation, Missense Myotonia Myotonia - genetics Myotonia - physiopathology Myotonia Congenita - genetics Myotonia Congenita - physiopathology Myotonic Disorders - genetics Myotonic Disorders - physiopathology NAV1.4 Voltage-Gated Sodium Channel - genetics Paralysis, Hyperkalemic Periodic - genetics Paralysis, Hyperkalemic Periodic - physiopathology Paramyotonia congenita Physical Exertion Sodium channel Zebrafish |
| title | A zebrafish model of nondystrophic myotonia with sodium channelopathy |
| url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-09-23T03%3A23%3A03IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20zebrafish%20model%20of%20nondystrophic%20myotonia%20with%20sodium%20channelopathy&rft.jtitle=Neuroscience%20letters&rft.au=Nam,%20Tai-Seung&rft.date=2020-01-01&rft.volume=714&rft.spage=134579&rft.epage=134579&rft.pages=134579-134579&rft.artnum=134579&rft.issn=0304-3940&rft.eissn=1872-7972&rft_id=info:doi/10.1016/j.neulet.2019.134579&rft_dat=%3Cproquest_cross%3E2310721801%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c362t-45008bd16e94057684d00cac657d2581b576ca90bbb4d569a20320d08a2013c53%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=2310721801&rft_id=info:pmid/31669315&rfr_iscdi=true |