Lipopolysaccharide-Induced Differential Expression of miRNAs in Male and Female Rhipicephalus haemaphysaloides Ticks
Lipopolysaccharide (LPS) stimulates the innate immune response in arthropods. In tick vectors, LPS activates expression of immune genes, including those for antibacterial peptides. miRNAs are 21-24 nt non-coding small RNAs that regulate target mRNAs at the post-transcriptional level. However, our un...
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| description | Lipopolysaccharide (LPS) stimulates the innate immune response in arthropods. In tick vectors, LPS activates expression of immune genes, including those for antibacterial peptides. miRNAs are 21-24 nt non-coding small RNAs that regulate target mRNAs at the post-transcriptional level. However, our understanding of tick innate immunity is limited to a few cellular immune reactions and some characterized immune molecules. Moreover, there is little information on the regulation of the immune system in ticks by miRNA. Therefore, this study aimed to analyze the differential expression of miRNAs in male and female ticks after LPS injection. LPS was injected into male and female Rhipicephalus haemaphysaloides ticks to stimulate immune response, with phosphate buffered saline (PBS)-injected ticks as negative controls. miRNAs from each group were sequenced and analyzed. In the PBS- and LPS-injected female ticks, 11.46 and 12.82 million reads of 18-30 nt were obtained respectively. There were 13.92 and 15.29 million reads of 18-30 nt obtained in the PBS- and LPS-injected male ticks, respectively. Expression of miRNAs in male ticks was greater than that in female ticks. There were 955 and 984 conserved miRNA families in the PBS- and LPS-injected female ticks, respectively, and correspondingly 1684 and 1552 conserved miRNA families in male ticks. Nine novel miRNAs were detected as common miRNAs in two or more tested samples. There were 37 known miRNAs up-regulated >10-fold and 33 down-regulated >10-fold in LPS-injected female ticks; and correspondingly 52 and 59 miRNAs in male ticks. Differential expression of miRNAs in PBS- and LPS-injected samples supports their involvement in the regulation of innate immunity. These data provide an important resource for more detailed functional analysis of miRNAs in this species. |
| doi_str_mv | 10.1371/journal.pone.0139241 |
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In tick vectors, LPS activates expression of immune genes, including those for antibacterial peptides. miRNAs are 21-24 nt non-coding small RNAs that regulate target mRNAs at the post-transcriptional level. However, our understanding of tick innate immunity is limited to a few cellular immune reactions and some characterized immune molecules. Moreover, there is little information on the regulation of the immune system in ticks by miRNA. Therefore, this study aimed to analyze the differential expression of miRNAs in male and female ticks after LPS injection. LPS was injected into male and female Rhipicephalus haemaphysaloides ticks to stimulate immune response, with phosphate buffered saline (PBS)-injected ticks as negative controls. miRNAs from each group were sequenced and analyzed. In the PBS- and LPS-injected female ticks, 11.46 and 12.82 million reads of 18-30 nt were obtained respectively. There were 13.92 and 15.29 million reads of 18-30 nt obtained in the PBS- and LPS-injected male ticks, respectively. Expression of miRNAs in male ticks was greater than that in female ticks. There were 955 and 984 conserved miRNA families in the PBS- and LPS-injected female ticks, respectively, and correspondingly 1684 and 1552 conserved miRNA families in male ticks. Nine novel miRNAs were detected as common miRNAs in two or more tested samples. There were 37 known miRNAs up-regulated >10-fold and 33 down-regulated >10-fold in LPS-injected female ticks; and correspondingly 52 and 59 miRNAs in male ticks. Differential expression of miRNAs in PBS- and LPS-injected samples supports their involvement in the regulation of innate immunity. These data provide an important resource for more detailed functional analysis of miRNAs in this species.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0139241</identifier><identifier>PMID: 26430879</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Acari ; Agriculture ; Analysis ; Animals ; Arthropoda ; Arthropods ; Boophilus ; Enzymes ; Expression vectors ; Female ; Functional analysis ; Gene expression ; Genetic aspects ; Genomics ; Haemaphysalis longicornis ; Identification ; Immune response ; Immune system ; Immunity ; Innate immunity ; Insects ; Kinases ; Laboratories ; Lipopolysaccharides ; Lipopolysaccharides - pharmacology ; Male ; MicroRNA ; MicroRNAs ; MicroRNAs - chemistry ; MicroRNAs - genetics ; miRNA ; Nucleic Acid Conformation ; Parasitology ; Peptides ; Physiological aspects ; Post-transcription ; Rhipicephalus ; Rhipicephalus - genetics ; Rhipicephalus haemaphysaloides ; RNA polymerase ; Ticks</subject><ispartof>PloS one, 2015-10, Vol.10 (10), p.e0139241-e0139241</ispartof><rights>COPYRIGHT 2015 Public Library of Science</rights><rights>2015 Wang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2015 Wang et al 2015 Wang et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-eca42f06f3a0c9e5e0b4d1846cc5111919298cb380b865def46356637a673d613</citedby><cites>FETCH-LOGICAL-c692t-eca42f06f3a0c9e5e0b4d1846cc5111919298cb380b865def46356637a673d613</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4592253/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4592253/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79343,79344</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26430879$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Munderloh, Ulrike Gertrud</contributor><creatorcontrib>Wang, Fangfang</creatorcontrib><creatorcontrib>Gong, Haiyan</creatorcontrib><creatorcontrib>Zhang, Houshuang</creatorcontrib><creatorcontrib>Zhou, Yongzhi</creatorcontrib><creatorcontrib>Cao, Jie</creatorcontrib><creatorcontrib>Zhou, Jinlin</creatorcontrib><title>Lipopolysaccharide-Induced Differential Expression of miRNAs in Male and Female Rhipicephalus haemaphysaloides Ticks</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Lipopolysaccharide (LPS) stimulates the innate immune response in arthropods. In tick vectors, LPS activates expression of immune genes, including those for antibacterial peptides. miRNAs are 21-24 nt non-coding small RNAs that regulate target mRNAs at the post-transcriptional level. However, our understanding of tick innate immunity is limited to a few cellular immune reactions and some characterized immune molecules. Moreover, there is little information on the regulation of the immune system in ticks by miRNA. Therefore, this study aimed to analyze the differential expression of miRNAs in male and female ticks after LPS injection. LPS was injected into male and female Rhipicephalus haemaphysaloides ticks to stimulate immune response, with phosphate buffered saline (PBS)-injected ticks as negative controls. miRNAs from each group were sequenced and analyzed. In the PBS- and LPS-injected female ticks, 11.46 and 12.82 million reads of 18-30 nt were obtained respectively. There were 13.92 and 15.29 million reads of 18-30 nt obtained in the PBS- and LPS-injected male ticks, respectively. Expression of miRNAs in male ticks was greater than that in female ticks. There were 955 and 984 conserved miRNA families in the PBS- and LPS-injected female ticks, respectively, and correspondingly 1684 and 1552 conserved miRNA families in male ticks. Nine novel miRNAs were detected as common miRNAs in two or more tested samples. There were 37 known miRNAs up-regulated >10-fold and 33 down-regulated >10-fold in LPS-injected female ticks; and correspondingly 52 and 59 miRNAs in male ticks. Differential expression of miRNAs in PBS- and LPS-injected samples supports their involvement in the regulation of innate immunity. These data provide an important resource for more detailed functional analysis of miRNAs in this species.</description><subject>Acari</subject><subject>Agriculture</subject><subject>Analysis</subject><subject>Animals</subject><subject>Arthropoda</subject><subject>Arthropods</subject><subject>Boophilus</subject><subject>Enzymes</subject><subject>Expression vectors</subject><subject>Female</subject><subject>Functional analysis</subject><subject>Gene expression</subject><subject>Genetic aspects</subject><subject>Genomics</subject><subject>Haemaphysalis longicornis</subject><subject>Identification</subject><subject>Immune response</subject><subject>Immune system</subject><subject>Immunity</subject><subject>Innate immunity</subject><subject>Insects</subject><subject>Kinases</subject><subject>Laboratories</subject><subject>Lipopolysaccharides</subject><subject>Lipopolysaccharides - pharmacology</subject><subject>Male</subject><subject>MicroRNA</subject><subject>MicroRNAs</subject><subject>MicroRNAs - chemistry</subject><subject>MicroRNAs - genetics</subject><subject>miRNA</subject><subject>Nucleic Acid Conformation</subject><subject>Parasitology</subject><subject>Peptides</subject><subject>Physiological aspects</subject><subject>Post-transcription</subject><subject>Rhipicephalus</subject><subject>Rhipicephalus - genetics</subject><subject>Rhipicephalus haemaphysaloides</subject><subject>RNA polymerase</subject><subject>Ticks</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk1Fv0zAQxyMEYqPwDRBEQkLw0GLHiZO8TKrGBpUKk8rg1XLsc-PixMFO0PbtcWk2NWgPyA-2Lr_7-_L3XRS9xGiBSY4_7OzgWm4WnW1hgTApkxQ_ik5xSZI5TRB5fHQ-iZ55v0MoIwWlT6OThKYEFXl5GvVr3dnOmlvPhai50xLmq1YOAmT8USsFDtpecxNf3HQOvNe2ja2KG735uvSxbuMv3EDMWxlfQrM_bmrdaQFdzc3g45qHaFcHdWODtI-vtfjpn0dPFDceXoz7LPp-eXF9_nm-vvq0Ol-u54KWST8HwdNEIaoIR6KEDFCVSlykVIgMY1ziMikLUZECVQXNJKiUkoxSknOaE0kxmUWvD7qdsZ6NhnmG82BMngbPArE6ENLyHeucbri7ZZZr9jdg3ZZx12thgGWShHIwzirOU4yAU66gqFCpMFaFzILW2XjbUDUgRTDOcTMRnX5pdc229jdLszJJsn0x70YBZ38N4HvWaC_AGN6CHQ51hwoCGdA3_6AP_91IbcPLMN0qG-4Ve1G2DA2QJUUR2Fm0eIAKS0KjRegupUN8kvB-khCYHm76LR-8Z6tvm_9nr35M2bdHbA3c9LW3ZuhD0_kpmB5A4az3DtS9yRix_XDcucH2w8HG4Qhpr44f6D7pbhrIH2a0Cjw</recordid><startdate>20151002</startdate><enddate>20151002</enddate><creator>Wang, Fangfang</creator><creator>Gong, Haiyan</creator><creator>Zhang, Houshuang</creator><creator>Zhou, Yongzhi</creator><creator>Cao, Jie</creator><creator>Zhou, Jinlin</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20151002</creationdate><title>Lipopolysaccharide-Induced Differential Expression of miRNAs in Male and Female Rhipicephalus haemaphysaloides Ticks</title><author>Wang, Fangfang ; Gong, Haiyan ; Zhang, Houshuang ; Zhou, Yongzhi ; Cao, Jie ; Zhou, Jinlin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-eca42f06f3a0c9e5e0b4d1846cc5111919298cb380b865def46356637a673d613</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Acari</topic><topic>Agriculture</topic><topic>Analysis</topic><topic>Animals</topic><topic>Arthropoda</topic><topic>Arthropods</topic><topic>Boophilus</topic><topic>Enzymes</topic><topic>Expression vectors</topic><topic>Female</topic><topic>Functional analysis</topic><topic>Gene expression</topic><topic>Genetic aspects</topic><topic>Genomics</topic><topic>Haemaphysalis longicornis</topic><topic>Identification</topic><topic>Immune response</topic><topic>Immune system</topic><topic>Immunity</topic><topic>Innate immunity</topic><topic>Insects</topic><topic>Kinases</topic><topic>Laboratories</topic><topic>Lipopolysaccharides</topic><topic>Lipopolysaccharides - pharmacology</topic><topic>Male</topic><topic>MicroRNA</topic><topic>MicroRNAs</topic><topic>MicroRNAs - chemistry</topic><topic>MicroRNAs - genetics</topic><topic>miRNA</topic><topic>Nucleic Acid Conformation</topic><topic>Parasitology</topic><topic>Peptides</topic><topic>Physiological aspects</topic><topic>Post-transcription</topic><topic>Rhipicephalus</topic><topic>Rhipicephalus - genetics</topic><topic>Rhipicephalus haemaphysaloides</topic><topic>RNA polymerase</topic><topic>Ticks</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Fangfang</creatorcontrib><creatorcontrib>Gong, Haiyan</creatorcontrib><creatorcontrib>Zhang, Houshuang</creatorcontrib><creatorcontrib>Zhou, Yongzhi</creatorcontrib><creatorcontrib>Cao, Jie</creatorcontrib><creatorcontrib>Zhou, Jinlin</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Opposing Viewpoints in Context (Gale)</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Fangfang</au><au>Gong, Haiyan</au><au>Zhang, Houshuang</au><au>Zhou, Yongzhi</au><au>Cao, Jie</au><au>Zhou, Jinlin</au><au>Munderloh, Ulrike Gertrud</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Lipopolysaccharide-Induced Differential Expression of miRNAs in Male and Female Rhipicephalus haemaphysaloides Ticks</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2015-10-02</date><risdate>2015</risdate><volume>10</volume><issue>10</issue><spage>e0139241</spage><epage>e0139241</epage><pages>e0139241-e0139241</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Lipopolysaccharide (LPS) stimulates the innate immune response in arthropods. In tick vectors, LPS activates expression of immune genes, including those for antibacterial peptides. miRNAs are 21-24 nt non-coding small RNAs that regulate target mRNAs at the post-transcriptional level. However, our understanding of tick innate immunity is limited to a few cellular immune reactions and some characterized immune molecules. Moreover, there is little information on the regulation of the immune system in ticks by miRNA. Therefore, this study aimed to analyze the differential expression of miRNAs in male and female ticks after LPS injection. LPS was injected into male and female Rhipicephalus haemaphysaloides ticks to stimulate immune response, with phosphate buffered saline (PBS)-injected ticks as negative controls. miRNAs from each group were sequenced and analyzed. In the PBS- and LPS-injected female ticks, 11.46 and 12.82 million reads of 18-30 nt were obtained respectively. There were 13.92 and 15.29 million reads of 18-30 nt obtained in the PBS- and LPS-injected male ticks, respectively. Expression of miRNAs in male ticks was greater than that in female ticks. There were 955 and 984 conserved miRNA families in the PBS- and LPS-injected female ticks, respectively, and correspondingly 1684 and 1552 conserved miRNA families in male ticks. Nine novel miRNAs were detected as common miRNAs in two or more tested samples. There were 37 known miRNAs up-regulated >10-fold and 33 down-regulated >10-fold in LPS-injected female ticks; and correspondingly 52 and 59 miRNAs in male ticks. Differential expression of miRNAs in PBS- and LPS-injected samples supports their involvement in the regulation of innate immunity. These data provide an important resource for more detailed functional analysis of miRNAs in this species.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>26430879</pmid><doi>10.1371/journal.pone.0139241</doi><oa>free_for_read</oa></addata></record> |
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| source | Public Library of Science (PLoS) Journals Open Access; MEDLINE; PMC (PubMed Central); DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Free Full-Text Journals in Chemistry |
| subjects | Acari Agriculture Analysis Animals Arthropoda Arthropods Boophilus Enzymes Expression vectors Female Functional analysis Gene expression Genetic aspects Genomics Haemaphysalis longicornis Identification Immune response Immune system Immunity Innate immunity Insects Kinases Laboratories Lipopolysaccharides Lipopolysaccharides - pharmacology Male MicroRNA MicroRNAs MicroRNAs - chemistry MicroRNAs - genetics miRNA Nucleic Acid Conformation Parasitology Peptides Physiological aspects Post-transcription Rhipicephalus Rhipicephalus - genetics Rhipicephalus haemaphysaloides RNA polymerase Ticks |
| title | Lipopolysaccharide-Induced Differential Expression of miRNAs in Male and Female Rhipicephalus haemaphysaloides Ticks |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-01T00%3A03%3A01IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Lipopolysaccharide-Induced%20Differential%20Expression%20of%20miRNAs%20in%20Male%20and%20Female%20Rhipicephalus%20haemaphysaloides%20Ticks&rft.jtitle=PloS%20one&rft.au=Wang,%20Fangfang&rft.date=2015-10-02&rft.volume=10&rft.issue=10&rft.spage=e0139241&rft.epage=e0139241&rft.pages=e0139241-e0139241&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0139241&rft_dat=%3Cgale_plos_%3EA430528819%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1719374013&rft_id=info:pmid/26430879&rft_galeid=A430528819&rft_doaj_id=oai_doaj_org_article_5d3a42115baa410ea6afe8b09f11f8d5&rfr_iscdi=true |