{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T18:00:56Z","timestamp":1789840856593,"version":"4.0.1"},"reference-count":56,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2020,9,10]],"date-time":"2020-09-10T00:00:00Z","timestamp":1599696000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100007161","name":"Boston University","doi-asserted-by":"publisher","award":["no number"],"award-info":[{"award-number":["no number"]}],"id":[{"id":"10.13039\/100007161","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Dengue is the most burdensome vector-borne viral disease in the world. Dengue virus (DENV), the etiological cause of dengue, is transmitted primarily by the Aedes aegypti mosquito. Like any arbovirus, the transmission cycle of dengue involves the complex interactions of a multitude of human and mosquito factors. One point during this transmission cycle that is rich in these interactions is the biting event by the mosquito, upon which its saliva is injected into the host. A number of components in mosquito saliva have been shown to play a pivotal role in the transmission of dengue, however one such component that is not as well characterized is extracellular vesicles. Here, using high-performance liquid chromatography in tandem with mass spectrometry, we show that dengue infection altered the protein cargo of Aedes aegypti extracellular vesicles, resulting in the packaging of proteins with infection-enhancing ability. Our results support the presence of an infection-dependent pro-viral protein packaging strategy that uses the differential packaging of pro-viral proteins in extracellular vesicles of Ae. aegypti saliva to promote transmission. These studies represent the first investigation into the function of Ae. aegypti extracellular vesicle cargo during dengue infection.<\/jats:p>","DOI":"10.3390\/ijms21186609","type":"journal-article","created":{"date-parts":[[2020,9,10]],"date-time":"2020-09-10T09:10:09Z","timestamp":1599729009000},"page":"6609","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Dengue Virus Infection of Aedes aegypti Alters Extracellular Vesicle Protein Cargo to Enhance Virus Transmission"],"prefix":"10.3390","volume":"21","author":[{"given":"Alexander S.","family":"Gold","sequence":"first","affiliation":[{"name":"Department of Microbiology, National Emerging Infectious Diseases Laboratories, Boston University School of Medicine, 620 Albany Street, Boston, MA 02118, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6971-4691","authenticated-orcid":false,"given":"Fabiana","family":"Feitosa-Suntheimer","sequence":"additional","affiliation":[{"name":"Department of Microbiology, National Emerging Infectious Diseases Laboratories, Boston University School of Medicine, 620 Albany Street, Boston, MA 02118, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3516-3358","authenticated-orcid":false,"given":"Ricardo V.","family":"Araujo","sequence":"additional","affiliation":[{"name":"Department of Microbiology, National Emerging Infectious Diseases Laboratories, Boston University School of Medicine, 620 Albany Street, Boston, MA 02118, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryan M.","family":"Hekman","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Boston University School of Medicine, 72 E. Concord Street, Boston, MA 02118, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sultan","family":"Asad","sequence":"additional","affiliation":[{"name":"Department of Microbiology, National Emerging Infectious Diseases Laboratories, Boston University School of Medicine, 620 Albany Street, Boston, MA 02118, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5956-5274","authenticated-orcid":false,"given":"Berlin","family":"Londono-Renteria","sequence":"additional","affiliation":[{"name":"Department of Entomology, Vector Biology Laboratory, Kansas State University, 1603 Old Claflin Pl, 123 Waters Hall, Manhattan, KS 66506, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew","family":"Emili","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Boston University School of Medicine, 72 E. Concord Street, Boston, MA 02118, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tonya M.","family":"Colpitts","sequence":"additional","affiliation":[{"name":"Department of Microbiology, National Emerging Infectious Diseases Laboratories, Boston University School of Medicine, 620 Albany Street, Boston, MA 02118, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,9,10]]},"reference":[{"key":"ref_1","unstructured":"Beatty, M.E., Letson, G.W., and Margolis, H.S. (2020, September 04). The Global Burden of Dengue. Available online: http:\/\/www.pdvi.org\/about_dengue\/GBD.asp."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"140004","DOI":"10.1038\/sdata.2014.4","article-title":"A global compendium of human dengue virus occurrence","volume":"1","author":"Messina","year":"2014","journal-title":"Sci. Data"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"712","DOI":"10.1016\/S1473-3099(16)00026-8","article-title":"The Global Burden of Dengue: An analysis from the Global Burden of Disease Study 2013","volume":"16","author":"Stanaway","year":"2016","journal-title":"Lancet Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e08347","DOI":"10.7554\/eLife.08347","article-title":"The global distribution of the arbovirus vectors Aedes aegypti and Ae. albopictus","volume":"4","author":"Kraemer","year":"2015","journal-title":"eLife"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Brady, O.J., Gething, P.W., Bhatt, S., Messina, J.P., Brownstein, J.S., Hoen, A.G., Moyes, C.L., Farlow, A.W., Scott, T.W., and Hay, S.I. (2012). Refining the Global Spatial Limits of Dengue Virus Transmission by Evidence-Based Consensus. PLoS Negl. Trop. Dis., 6.","DOI":"10.1371\/journal.pntd.0001760"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1038\/nature12060","article-title":"The global distribution and burden of dengue","volume":"496","author":"Bhatt","year":"2013","journal-title":"Nature"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1563","DOI":"10.1136\/bmj.324.7353.1563","article-title":"Dengue: An escalating problem","volume":"324","author":"Gibbons","year":"2002","journal-title":"BMJ"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1016\/S0188-4409(02)00378-8","article-title":"The Global emergence\/resurgence of arboviral diseases as public health problems","volume":"33","author":"Gubler","year":"2002","journal-title":"Arch. Med. Res."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Beatty, M.E., Stone, A., Fitzsimons, D.W., Hanna, J.N., Lam, S.K., Vong, S., Guzman, M.G., Mendez-Galvan, J.F., Halstead, S.B., and Letson, G.W. (2010). Best practices in dengue surveillance: A report from the Asia-Pacific and Americas Dengue Prevention Boards. PLoS Negl. Trop. Dis., 4.","DOI":"10.1371\/journal.pntd.0000890"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1097\/PEC.0b013e31828ed30e","article-title":"A review of dengue fever: A resurging tropical disease","volume":"29","author":"Mangold","year":"2013","journal-title":"Pediatr. Emerg. Care"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1508","DOI":"10.1038\/s41564-019-0476-8","article-title":"The current and future global distribution and population at risk of dengue","volume":"4","author":"Messina","year":"2019","journal-title":"Nat. Microbiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"S96","DOI":"10.1093\/infdis\/jiw423","article-title":"Current status of dengue therapeutics research and development","volume":"215","author":"Low","year":"2017","journal-title":"J. Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1007\/s00005-014-0324-8","article-title":"Transmission-blocking vaccines: Focus on anti-vector vaccines against tick-borne diseases","volume":"63","author":"Neelakanta","year":"2014","journal-title":"Arch. Immunol. Ther. Exp."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Styer, L.M., Kent, K.A., Albright, R.G., Bennett, C.J., Kramer, L.D., and Bernard, K.A. (2007). Mosquitoes Inoculate High Doses of West Nile Virus as They Probe and Feed on Live Hosts. PLoS Pathog., 3.","DOI":"10.1371\/journal.ppat.0030132"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1016\/j.trstmh.2008.01.024","article-title":"The enhancement of arbovirus transmission and disease by mosquito saliva is associated with modulation of the host immune response","volume":"102","author":"Schneider","year":"2008","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1517","DOI":"10.1128\/JVI.01112-10","article-title":"Mosquito Saliva Causes Enhancement of West Nile Virus Infection in Mice","volume":"85","author":"Styer","year":"2010","journal-title":"J. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1146\/annurev.ento.48.060402.102812","article-title":"Role of arthropod saliva in bloofeeding: Sialome and post-sialome perspectives","volume":"48","author":"Ribeiro","year":"2003","journal-title":"Annu. Rev. Entomol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1128\/JVI.02235-13","article-title":"Mosquito saliva serine protease enhances dissemination of dengue virus into the mammalian host","volume":"88","author":"Conway","year":"2013","journal-title":"J. Virol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Vogt, M.B., Lahon, A., Arya, R.P., Kneubehl, A.R., Spencer Clinton, J.L., Paust, S., and Rico-Hesse, R. (2018). Mosquito saliva alone has profound effects on the human immune system. PLoS Negl. Trop. Dis., 12.","DOI":"10.1371\/journal.pntd.0006439"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4141","DOI":"10.4049\/jimmunol.176.7.4141","article-title":"Mast cell-dependent down-regulation of antigen-specific immune responses by mosquito bites","volume":"176","author":"Depinay","year":"2006","journal-title":"J. Immunol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1159\/000371349","article-title":"Immunological aspects of the immune response induced by mosquito allergens","volume":"165","author":"Cantillo","year":"2014","journal-title":"Int. Arch. Allergy Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Conway, M.J., Londono-Renteria, B., Troupin, A., Watson, A.M., Klimstra, W.B., Fikrig, E., and Colpitts, T.M. (2016). Aedes aegypti D7 Saliva protein inhibits dengue virus infection. PLoS Negl. Trop. Dis., 10.","DOI":"10.1371\/journal.pntd.0004941"},{"key":"ref_23","first-page":"1","article-title":"A mosquito salivary protein promotes flavivirus transmission by activation of autophagy","volume":"11","author":"Sun","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1038\/nrm.2017.125","article-title":"Shedding light on the cell biology of extracellular vesicles","volume":"19","author":"Raposo","year":"2018","journal-title":"Nat. Rev. Mol. Cell Boil."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1038\/s41580-019-0158-7","article-title":"Extracellular vesicles: A new communication paradigm?","volume":"20","author":"Raposo","year":"2019","journal-title":"Nat. Rev. Mol. Cell Boil."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Urbanelli, L., Buratta, S., Tancini, B., Sagini, K., Delo, F., Porcellati, S., and Emiliani, C. (2019). The Role of Extracellular Vesicles in Viral Infection and Transmission. Vaccines, 7.","DOI":"10.3390\/vaccines7030102"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"E6604","DOI":"10.1073\/pnas.1720125115","article-title":"Arthropod EVs mediate dengue virus transmission through interaction with a tetraspanin domain containing glycoprotein Tsp29Fb","volume":"115","author":"Vora","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Zhou, W., Woodson, M., Neupane, B., Bai, F., Sherman, M.B., Choi, K.H., Neelakanta, G., and Sultana, H. (2018). Exosomes serve as novel modes of tick-borne flavivirus transmission from arthropod to human cells and facilitates dissemination of viral RNA and proteins to the vertebrate neuronal cells. PLoS Pathog., 14.","DOI":"10.1371\/journal.ppat.1006764"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.virusres.2019.03.015","article-title":"Isolation and characterization of exosomes released from mosquito cells infected with dengue virus","volume":"266","author":"Medina","year":"2019","journal-title":"Virus Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1016\/j.pt.2018.04.001","article-title":"Exosome-Mediated Pathogen Transmission by Arthropod Vectors","volume":"34","author":"Hackenberg","year":"2018","journal-title":"Trends Parasitol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1792","DOI":"10.3389\/fmicb.2018.01792","article-title":"Characterization of Dendritic Cell-Derived Extracellular Vesicles During Dengue Virus Infection","volume":"9","author":"Martins","year":"2018","journal-title":"Front. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1038\/jid.2013.251","article-title":"Aedes aegypti Saliva Contains a Prominent 34-kDa Protein that Strongly Enhances Dengue Virus Replication in Human Keratinocytes","volume":"134","author":"Surasombatpattana","year":"2014","journal-title":"J. Investig. Dermatol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1128\/MMBR.00059-12","article-title":"The Unexpected Roles of Eukaryotic Translation Elongation Factors in RNA Virus Replication and Pathogenesis","volume":"77","author":"Li","year":"2013","journal-title":"Microbiol. Mol. Boil. Rev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"6433","DOI":"10.1128\/jvi.71.9.6433-6444.1997","article-title":"Translation Elongation Factor-1 Alpha Interacts with the 39 Stem-Loop Region of West Nile Virus Genomic RNA","volume":"71","author":"Blackwell","year":"1997","journal-title":"J. Virol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"10172","DOI":"10.1128\/JVI.00531-07","article-title":"Interaction between the Cellular Protein eEF1A and the 3\u2032-Terminal Stem-Loop of West Nile Virus Genomic RNA Facilitates Viral Minus-Strand RNA Synthesis","volume":"81","author":"Davis","year":"2007","journal-title":"J. Virol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1006\/viro.2002.1407","article-title":"Translation Elongation Factor-1\u03b1, La, and PTB Interact with the 3\u2032 Untranslated Region of Dengue 4 Virus RNA","volume":"295","year":"2002","journal-title":"Virology"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Raquin, V., Merkling, S.H., Gausson, V., Moltini-Conclois, I., Frangeul, L., Varet, H., Dillies, M.-A., Saleh, M.-C., and Lambrechts, L. (2017). Individual co-variation between viral RNA load and gene expression reveals novel host factors during early dengue virus infection of the Aedes aegypti midgut. PLoS Negl. Trop. Dis., 11.","DOI":"10.1371\/journal.pntd.0006152"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1108","DOI":"10.1016\/j.cell.2015.10.046","article-title":"Defining Hsp70 Subnetworks in Dengue Virus Replication Reveals Key Vulnerability in Flavivirus Infection","volume":"163","author":"Taguwa","year":"2015","journal-title":"Cell"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Asea, A., and Kaur, P. (2018). Heat Shock Protein 70 (HSP70) Family in Dengue Virus Infection. Regulation of Heat Shock Protein Responses. Heat Shock Proteins, Springer.","DOI":"10.1007\/978-3-319-74715-6"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1080\/22221751.2018.1557988","article-title":"Heat shock protein 70 (Hsp70) mediates Zika virus entry, replication, and egress from host cells","volume":"8","author":"Pujhari","year":"2019","journal-title":"Emerg. Microbes Infect."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1718","DOI":"10.1126\/science.1138878","article-title":"Genome Sequence of Aedes aegypti, a Major Arbovirus Vector","volume":"316","author":"Nene","year":"2007","journal-title":"Science"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/S0022-1910(00)00096-2","article-title":"Protein catabolism in mosquitoes: Ureotely and uricotely in larval and imaginal Aedes aegypti","volume":"47","author":"Briegel","year":"2001","journal-title":"J. Insect Physiol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Isoe, J., and Scaraffia, P.Y. (2013). Urea Synthesis and Excretion in Aedes aegypti Mosquitoes Are Regulated by a Unique Cross-Talk Mechanism. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0065393"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"638","DOI":"10.1111\/j.1476-5381.2009.00291.x","article-title":"Arginase: An emerging key player in the mammalian immune system","volume":"158","author":"Munder","year":"2009","journal-title":"Br. J. Pharmacol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1146\/annurev.immunol.15.1.323","article-title":"Nitric oxide and macrophage function","volume":"15","author":"MacMicking","year":"1997","journal-title":"Annu. Rev. Immunol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"5","DOI":"10.3389\/fimmu.2014.00428","article-title":"The Role of Myeloid Cell Activation and Arginine Metabolism in the Pathogenesis of Virus-Induced Diseases","volume":"5","author":"Burrack","year":"2014","journal-title":"Front. Immunol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"103581","DOI":"10.1016\/j.micpath.2019.103581","article-title":"An overview of the immune response and Arginase I on CHIKV immunopathogenesis","volume":"135","author":"Suzukawa","year":"2019","journal-title":"Microb. Pathog."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"2886","DOI":"10.1002\/ijc.24265","article-title":"Hepatitis C virus targets over-expression of arginase I in hepatocarcinogenesis","volume":"124","author":"Cao","year":"2009","journal-title":"Int. J. Cancer"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"3003","DOI":"10.1099\/vir.0.81880-0","article-title":"Antiviral action of nitric oxide on dengue virus type 2 replication","volume":"87","author":"Takhampunya","year":"2006","journal-title":"J. Gen. Virol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1111\/j.1574-695X.2009.00544.x","article-title":"Nitric oxide in dengue and dengue haemorrhagic fever: Necessity or nuisance?","volume":"56","author":"Chaturvedi","year":"2009","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"49","DOI":"10.2987\/8756-971X(2005)21[49:QOWNVI]2.0.CO;2","article-title":"Quantification of West Nile Virus in Vector Mosquito Saliva","volume":"21","author":"Colton","year":"2005","journal-title":"J. Am. Mosq. Control Assoc."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"108","DOI":"10.2987\/09-5935.1","article-title":"A Simple Method for Determining Arbovirus Transmission in Mosquitoes","volume":"26","author":"Anderson","year":"2010","journal-title":"Am. Mosq. Control Assoc."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1006\/viro.1997.8500","article-title":"Construction of Infectious cDNA Clones for Dengue 2 Virus: Strain 16681 and Its Attenuated Vaccine Derivative, Strain PDK-53","volume":"230","author":"Kinney","year":"1997","journal-title":"Virology"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1038\/nature04601","article-title":"Analysis of the DNA sequence and duplication history of human chromosome 15","volume":"440","author":"Zody","year":"2006","journal-title":"Nature"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"834","DOI":"10.3892\/ijmm.2017.3080","article-title":"Comparison of isolation methods of exosomes and exosomal RNA form cell culture medium and serum","volume":"40","author":"Tang","year":"2017","journal-title":"Int. J. Mol. Med."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Ribeiro, J.M.C., Arc\u00e0, B., Lombardo, F., Calvo, E., Phan, V.M., Chandra, P.K., and Wikel, S.K. (2007). An annotated catalogue of salivary gland transcripts in the adult female mosquito, Aedes \u00e6gypti. BMC Genom., 8.","DOI":"10.1186\/1471-2164-8-6"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/18\/6609\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:08:33Z","timestamp":1760177313000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/18\/6609"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,9,10]]},"references-count":56,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2020,9]]}},"alternative-id":["ijms21186609"],"URL":"https:\/\/doi.org\/10.3390\/ijms21186609","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,9,10]]}}}