{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,2]],"date-time":"2025-11-02T11:10:13Z","timestamp":1762081813931,"version":"build-2065373602"},"reference-count":60,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T00:00:00Z","timestamp":1671580800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Interdisciplinary Scientific and Educational School of Moscow State University \u201cPhotonic and Quantum Technologies. Digital Medicine\u201d and by M.V. Lomonosov Moscow State University state task","award":["121041500039-8"],"award-info":[{"award-number":["121041500039-8"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Gold nanoparticles (AuNPs) are popular labels for colorimetric detection of various analytes, involving proteins, nucleic acids, viruses, and whole cells because of their outstanding optical properties, inertness, and modification variability. In this work, we present an improved approach for enhancement of color intensity for DNA membrane microarrays based on seed-mediated growth of AuNP labels. Biotin-labeled DNA is hybridized with capture oligonucleotide probes immobilized on the microarrays. Then biotin is revealed by a streptavidin\u2013AuNP conjugate followed by the detection of AuNPs. Optimization of seed-mediated enlargement of AuNPs by the reduction of tetrachloroauric acid with hydroxylamine made it possible to change the coloring of specific spots on the microarrays from pink to a more contrasting black with minor background staining. Mean size of the resulting AuNPs was four times larger than before the enhancement. Adjusting the pH of HAuCl4 solution to 3.5 and use of a large excess of hydroxylamine increased the signal\/background ratio by several times. The method\u2019s applicability was demonstrated for quantification of a short oligonucleotide of 19 bases and full-length TEM-type \u03b2-lactamase genes of 860 bp responsible for the development of bacterial resistance against \u03b2-lactam antibiotics. Improved protocol for AuNP enlargement may be further transferred to any other membrane-based assays of nucleic acids with both instrumental and visual colorimetric detection.<\/jats:p>","DOI":"10.3390\/bios13010002","type":"journal-article","created":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T06:11:24Z","timestamp":1671603084000},"page":"2","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Improvement of Seed-Mediated Growth of Gold Nanoparticle Labels for DNA Membrane-Based Assays"],"prefix":"10.3390","volume":"13","author":[{"given":"Galina V.","family":"Presnova","sequence":"first","affiliation":[{"name":"Department of Chemistry, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4082-9684","authenticated-orcid":false,"given":"Gleb A.","family":"Zhdanov","sequence":"additional","affiliation":[{"name":"Department of Chemistry, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luibov Yu.","family":"Filatova","sequence":"additional","affiliation":[{"name":"Department of Chemistry, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mariya M.","family":"Ulyashova","sequence":"additional","affiliation":[{"name":"Department of Chemistry, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9213-0165","authenticated-orcid":false,"given":"Denis E.","family":"Presnov","sequence":"additional","affiliation":[{"name":"D.V. Skobeltsyn Institute of Nuclear Physics, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia"},{"name":"MSU Quantum Technology Centre, 119991 Moscow, Russia"},{"name":"Cryoelectronics Lab, Faculty of Physics, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4025-5967","authenticated-orcid":false,"given":"Maya Yu.","family":"Rubtsova","sequence":"additional","affiliation":[{"name":"Department of Chemistry, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Wang, W., Wang, X., Liu, J., Lin, C., Liu, J., and Wang, J. (2022). The integration of gold nanoparticles with polymerase chain reaction for constructing colorimetric sensing platforms for detection of health-related DNA and proteins. Biosensors, 12.","DOI":"10.3390\/bios12060421"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"878","DOI":"10.1007\/s13346-020-00771-y","article-title":"Perspectives of characterization and bioconjugation of gold nanoparticles and their application in lateral flow immunosensing","volume":"10","author":"Borse","year":"2020","journal-title":"Drug Deliv. Transl. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.bios.2015.05.050","article-title":"Nanoparticle-based lateral flow biosensors","volume":"73","year":"2015","journal-title":"Biosens. Bioelectron."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2439","DOI":"10.1007\/s00216-019-01685-6","article-title":"Colorimetric assay for ultrasensitive detection of Ag(I) ions based on the formation of gold nanoparticle oligomers","volume":"411","author":"Jiang","year":"2019","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"9637","DOI":"10.1039\/C4TC02034K","article-title":"Upconversion luminescence nanoparticles-based lateral flow immunochromatographic assay for cephalexin detection","volume":"2","author":"Liu","year":"2014","journal-title":"J. Mater. Chem. C."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"21535","DOI":"10.3390\/s141121535","article-title":"Magnetic bead-based colorimetric immunoassay for aflatoxin B1 using gold nanoparticles","volume":"14","author":"Wang","year":"2014","journal-title":"Sensors"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1016\/j.cclet.2016.01.013","article-title":"Fabrication and characterization of gold nano particles for DNA biosensor applications","volume":"27","author":"Mohammed","year":"2016","journal-title":"Chin. Chem. Lett."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Gribanyov, D., Zhdanov, G., Olenin, A., Lisichkin, G., Gambaryan, A., Kukushkin, V., and Zavyalova, E. (2021). SERS-based colloidal aptasensors for quantitative determination of influenza virus. Int. J. Mol Sci., 22.","DOI":"10.3390\/ijms22041842"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2291","DOI":"10.1007\/s00216-018-0850-z","article-title":"Functionalized Au@Ag-Au nanoparticles as an optical and SERS dual probe for lateral flow sensing","volume":"410","author":"Bai","year":"2018","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2981931","DOI":"10.1155\/2016\/2981931","article-title":"Nanotechnology-based surface plasmon resonance affinity biosensors for in vitro diagnostics","volume":"2016","author":"Antiochia","year":"2016","journal-title":"Int. J. Anal. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.talanta.2018.09.023","article-title":"Ultra-sensitive detection by metal nanoparticles-mediated enhanced SPR biosensors","volume":"192","author":"Fathi","year":"2019","journal-title":"Talanta"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"0094","DOI":"10.1021\/acsomega.9b00340","article-title":"Scanometric detection of tomato leaf curl New Delhi viral DNA using mono- and bifunctional AuNP-conjugated oligonucleotide probes","volume":"4","author":"Dharanivasan","year":"2019","journal-title":"ACS Omega"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1474","DOI":"10.1002\/tcr.202000087","article-title":"Functionalized gold nanoparticles: Synthesis. Properties and biomedical applications","volume":"20","author":"Fan","year":"2020","journal-title":"Chem. Rec."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1007\/978-1-0716-0319-2_26","article-title":"Gold Nanoclusters, Gold Nanoparticles, and Analytical Techniques for Their Characterization","volume":"2118","year":"2020","journal-title":"Methods Mol. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1039\/C7NR06367A","article-title":"Gold nanoparticle-based colorimetric biosensors","volume":"10","author":"Aldewachi","year":"2018","journal-title":"Nanoscale"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1016\/j.talanta.2015.10.068","article-title":"Silver and gold enhancement methods for lateral flow immunoassays","volume":"148","year":"2016","journal-title":"Talanta"},{"key":"ref_17","first-page":"57297","article-title":"Rapid nanoprobe signal enhancement by in situ gold nanoparticle synthesis","volume":"133","author":"Dias","year":"2018","journal-title":"J. Vis. Exp."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1007\/s40820-017-0180-2","article-title":"Hierarchical Nanogold labels to improve the sensitivity of lateral flow immunoassay","volume":"10","author":"Serebrennikova","year":"2018","journal-title":"Nano-Micro Lett."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1080\/05704928.2013.807817","article-title":"Silver Enhancement of gold nanoparticles for biosensing: From qualitative to quantitative","volume":"49","author":"Liu","year":"2014","journal-title":"Appl. Spectrosc. Rev."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"6620","DOI":"10.1021\/ac800834n","article-title":"Determination of size and concentration of gold nanoparticles from extinction spectra","volume":"80","author":"Khlebtsov","year":"2008","journal-title":"Anal. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"778269","DOI":"10.3389\/fbioe.2021.778269","article-title":"Sensitive colorimetric detection of interleukin-6 via Lateral low Assay incorporated silver amplification method","volume":"9","author":"Rahbar","year":"2021","journal-title":"Front. Bioeng. Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1080\/09540105.2017.1401044","article-title":"Silver-enhanced lateral flow immunoassay for highly-sensitive detection of potato leafroll virus","volume":"29","author":"Panferov","year":"2018","journal-title":"Food Agricul. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"9859","DOI":"10.1007\/s00216-013-7428-6","article-title":"Increased sensitivity of lateral flow immunoassay for ochratoxin A through silver enhancement","volume":"405","author":"Anfossi","year":"2013","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1134\/S000368381505004X","article-title":"Development of an immunochromatographic test system for the detection of Helicobacter pylori antigens","volume":"51","author":"Byzova","year":"2015","journal-title":"Appl. Biochem. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Karimadom, B.R., and Kornweitz, H. (2021). Mechanism of Producing Metallic Nanoparticles, with an Emphasis on Silver and Gold Nanoparticles, Using Bottom-Up Methods. Molecules, 26.","DOI":"10.3390\/molecules26102968"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1126\/science.1089171","article-title":"A Hybridization model for the plasmon response of complex nanostructures","volume":"302","author":"Prodan","year":"2003","journal-title":"Science"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"24868","DOI":"10.1364\/OE.18.024868","article-title":"Huge light scattering from active anisotropic spherical particles","volume":"18","author":"Fan","year":"2010","journal-title":"Opt. Express"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.nantod.2015.02.007","article-title":"Strategies for enhancing the sensitivity of plasmonic nanosensors","volume":"10","author":"Guo","year":"2015","journal-title":"Nano Today"},{"key":"ref_29","first-page":"e53598","article-title":"Multifunctional hybrid Fe2O3-Au nanoparticles for efficient plasmonic heating","volume":"108","author":"Murph","year":"2016","journal-title":"J. Vis. Exp."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4796","DOI":"10.1039\/b809585j","article-title":"Preparation of high quality nanowires by tobacco mosaic virus templating of gold nanoparticles","volume":"18","author":"Bromley","year":"2008","journal-title":"J. Mater. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1911","DOI":"10.1166\/jnn.2007.740","article-title":"Synthesis of Au nanoparticles at \u201call\u201d pH by H2O2 reduction of HAuCl","volume":"7","author":"Panda","year":"2007","journal-title":"J. Nanosci. Nanotechnol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1373\/clinchem.2007.091801","article-title":"Nanogold catalysis\u2013based immunoresonance-scattering spectral assay for trace complement component","volume":"54","author":"Jiang","year":"2008","journal-title":"Clin. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3227","DOI":"10.1021\/ac500381e","article-title":"DNA\u2013gold nanoparticle conjugates-based nanoplasmonic probe for specific differentiation of cell types","volume":"86","author":"Yang","year":"2014","journal-title":"Anal. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3135","DOI":"10.1007\/s00604-016-1969-2","article-title":"Colorimetric detection of lysozyme based on its effect on the growth of gold nanoparticles induced by the reaction of chloroauric acid and hydroxylamine","volume":"183","author":"Wang","year":"2016","journal-title":"Microchim. Acta"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1016\/j.snb.2016.10.084","article-title":"Colorimetric detection of Hg2+ based on target-mediated growth of gold nanoparticles","volume":"241","author":"Zhao","year":"2017","journal-title":"Sens. Actuators B Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1807","DOI":"10.1007\/s00216-018-1450-7","article-title":"MicroRNA sensors based on gold nanoparticles","volume":"411","author":"Coutinho","year":"2019","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_37","first-page":"247","article-title":"Array-based nano-amplification technique was applied in detection of hepatitis E virus","volume":"39","author":"Liu","year":"2006","journal-title":"J. Biochem. Mol. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"5126","DOI":"10.1021\/la200150h","article-title":"pH Dependence of the size and crystallographic orientation of the gold nanoparticles prepared by seed-mediated growth","volume":"27","author":"Rahman","year":"2011","journal-title":"Langmuir"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.jcis.2005.06.012","article-title":"Formation mechanism of nonspherical gold nanoparticles during seeding growth: Roles of anion adsorption and reduction rate","volume":"293","author":"Cao","year":"2006","journal-title":"J. Colloid Interface Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2258","DOI":"10.1021\/nn305395p","article-title":"pH-Dependent Evolution of Five-Star Gold Nanostructures: An Experimental and Computational Study","volume":"7","author":"Wang","year":"2013","journal-title":"ACS Nano"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e01076-e18","DOI":"10.1128\/AAC.01076-18","article-title":"Past and present perspectives of \u03b2-lactamases","volume":"62","author":"Bush","year":"2018","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1252","DOI":"10.1016\/j.bios.2010.06.053","article-title":"Oligonucleotide microarrays with horseradish peroxidase-based detection for the identification of extended-spectrum \u03b2-lactamases","volume":"26","author":"Rubtsova","year":"2010","journal-title":"Biosens. Bioelectron."},{"key":"ref_43","first-page":"579","article-title":"Quantitative lateral flow immunoassay for total prostate specific antigen in serum","volume":"494","author":"Andreeva","year":"2015","journal-title":"Anal. Lett."},{"key":"ref_44","first-page":"164","article-title":"and Egorov, A.M. Streptavidin conjugates with gold nanoparticles for visualization of single DNA interactions on the silicon surface","volume":"8","author":"Presnova","year":"2014","journal-title":"Biochem. Suppl. Ser. B Biomed. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Presnova, G.V., Presnov, D.E., Filippova, A.A., Tsiniaikin, I.I., Ulyashova, M.M., and Rubtsova, M.Y. (2022). Multiplex digital quantification of \u03b2-lactamase genes in antibiotic-resistant bacteria by counting gold nanoparticle labels on silicon microchips. Biosensors, 12.","DOI":"10.3390\/bios12040226"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"7238","DOI":"10.1021\/jp057170o","article-title":"Calculated absorption and scattering properties of gold nanoparticles of different size. Shape and composition:\u2009Applications in biological imaging and biomedicine","volume":"110","author":"Jain","year":"2006","journal-title":"J. Phys. Chem. B"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"10779","DOI":"10.1021\/la503209b","article-title":"Little adjustments significantly improve the Turkevich synthesis of gold nanoparticles","volume":"30","author":"Schulz","year":"2014","journal-title":"Langmuir"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1134\/S1995078018060095","article-title":"Study of growth of bare and protein-modified gold nanoparticles in the presence of hydroxylamine and tetrachloroaurate","volume":"13","author":"Panferov","year":"2018","journal-title":"Nanotechnol. Russ."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1016\/j.jconrel.2016.06.017","article-title":"DLS and zeta potential\u2014What they are and what they are not?","volume":"235","author":"Bhattacharjee","year":"2016","journal-title":"J. Control. Release"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.aca.2013.04.022","article-title":"Gold immunochromatographic strips for enhanced detection of Avian influenza and Newcastle disease viruses","volume":"782","author":"Li","year":"2013","journal-title":"Anal. Chim. Acta"},{"key":"ref_51","first-page":"87","article-title":"Bacterial TEM type beta-lactamases: Structure and analysis of mutations","volume":"12","author":"Grigorenko","year":"2018","journal-title":"Biochem. Suppl B Biomed. Chem."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1503","DOI":"10.1126\/science.1067003","article-title":"Array-Based Electrical Detection of DNA with Nanoparticle Probes","volume":"295","author":"Park","year":"2002","journal-title":"Science"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"3315","DOI":"10.1039\/b926940a","article-title":"Label-free and naked eye detection of PNA\/DNA hybridization using enhancement of gold nanoparticles","volume":"46","author":"Kim","year":"2010","journal-title":"Chem. Commun."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"7601","DOI":"10.1039\/c2cc32869k","article-title":"Scanometric analysis of DNA microarrays using DNA intercalator-conjugated gold nanoparticles","volume":"48","author":"Cho","year":"2012","journal-title":"Chem. Commun."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"4732","DOI":"10.1039\/c1an15610a","article-title":"Electrochemical DNA sensor by the assembly of graphene and DNA-conjugated gold nanoparticles with silver enhancement strategy","volume":"136","author":"Lin","year":"2011","journal-title":"Analyst"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1935","DOI":"10.1007\/s00216-012-6297-8","article-title":"Novel plastic biochips for colorimetric detection of biomolecules","volume":"404","author":"Wen","year":"2012","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"656","DOI":"10.1007\/s11172-011-0101-3","article-title":"Colorimetric detection of immobilized horseradish peroxidase based on the co-oxidation of benzidine derivatives and 4-chloro-1-naphthol","volume":"60","author":"Ulyashova","year":"2011","journal-title":"Russ. Chem. Bull."},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Kanokudom, S., Assawakongkarat, T., Akeda, Y., Ratthawongjirakul, P., Chuanchuen, R., and Chaichanawongsaroj, N. (2021). Rapid detection of extended spectrum \u03b2-lactamase producing Escherichia coli isolated from fresh pork meat and pig cecum samples using multiplex recombinase polymerase amplification and lateral flow strip analysis. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0248536"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1039\/C2CC37037A","article-title":"Long genomic DNA amplicons adsorption onto unmodified gold nanoparticles for colorimetric detection of Bacillus anthracis","volume":"49","author":"Deng","year":"2013","journal-title":"Chem. Commun."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.bios.2016.09.031","article-title":"Denaturation strategies for detection of double stranded PCR products on GMR magnetic biosensor array","volume":"93","author":"Rizzi","year":"2017","journal-title":"Biosens. Bioelectron."}],"container-title":["Biosensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6374\/13\/1\/2\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:45:23Z","timestamp":1760147123000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6374\/13\/1\/2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,21]]},"references-count":60,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["bios13010002"],"URL":"https:\/\/doi.org\/10.3390\/bios13010002","relation":{},"ISSN":["2079-6374"],"issn-type":[{"type":"electronic","value":"2079-6374"}],"subject":[],"published":{"date-parts":[[2022,12,21]]}}}