{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T17:03:05Z","timestamp":1774976585242,"version":"3.50.1"},"reference-count":48,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2022,12,16]],"date-time":"2022-12-16T00:00:00Z","timestamp":1671148800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Fonds Wetenschappelijk Onderzoek Vlaanderen","award":["1501219N"],"award-info":[{"award-number":["1501219N"]}]},{"name":"Fonds Wetenschappelijk Onderzoek Vlaanderen","award":["G065421N"],"award-info":[{"award-number":["G065421N"]}]},{"name":"Fonds Wetenschappelijk Onderzoek Vlaanderen","award":["TT-ZAPBOF 33049"],"award-info":[{"award-number":["TT-ZAPBOF 33049"]}]},{"name":"Fonds Wetenschappelijk Onderzoek Vlaanderen","award":["TOP-BOF 35017"],"award-info":[{"award-number":["TOP-BOF 35017"]}]},{"name":"University of Antwerp","award":["1501219N"],"award-info":[{"award-number":["1501219N"]}]},{"name":"University of Antwerp","award":["G065421N"],"award-info":[{"award-number":["G065421N"]}]},{"name":"University of Antwerp","award":["TT-ZAPBOF 33049"],"award-info":[{"award-number":["TT-ZAPBOF 33049"]}]},{"name":"University of Antwerp","award":["TOP-BOF 35017"],"award-info":[{"award-number":["TOP-BOF 35017"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The application of in vivo bioluminescent imaging in infectious disease research has significantly increased over the past years. The detection of transgenic parasites expressing wildtype firefly luciferase is however hampered by a relatively low and heterogeneous tissue penetrating capacity of emitted light. Solutions are sought by using codon-optimized red-shifted luciferases that yield higher expression levels and produce relatively more red or near-infrared light, or by using modified bioluminescent substrates with enhanced cell permeability and improved luminogenic or pharmacokinetic properties. In this study, the in vitro and in vivo efficacy of two modified bioluminescent substrates, CycLuc1 and AkaLumine-HCl, were compared with that of D-luciferin as a gold standard. Comparisons were made in experimental and insect-transmitted animal models of leishmaniasis (caused by intracellular Leishmania species) and African trypanosomiasis (caused by extracellular Trypanosoma species), using parasite strains expressing the red-shifted firefly luciferase PpyRE9. Although the luminogenic properties of AkaLumine-HCl and D-luciferin for in vitro parasite detection were comparable at equal substrate concentrations, AkaLumine-HCl proved to be unsuitable for in vivo infection follow-up due to high background signals in the liver. CycLuc1 presented a higher in vitro luminescence compared to the other substrates and proved to be highly efficacious in vivo, even at a 20-fold lower dose than D-luciferin. This efficacy was consistent across infections with the herein included intracellular and extracellular parasitic organisms. It can be concluded that CycLuc1 is an excellent and broadly applicable alternative for D-luciferin, requiring significantly lower doses for in vivo bioluminescent imaging in rodent models of leishmaniasis and African trypanosomiasis.<\/jats:p>","DOI":"10.3390\/ijms232416074","type":"journal-article","created":{"date-parts":[[2022,12,19]],"date-time":"2022-12-19T05:25:46Z","timestamp":1671427546000},"page":"16074","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Comparison of Bioluminescent Substrates in Natural Infection Models of Neglected Parasitic Diseases"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1571-7296","authenticated-orcid":false,"given":"Sarah","family":"Hendrickx","sequence":"first","affiliation":[{"name":"Laboratory of Microbiology, Parasitology and Hygiene (LMPH), University of Antwerp, 2610 Wilrijk, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2783-4860","authenticated-orcid":false,"given":"Dimitri","family":"Bult\u00e9","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, Parasitology and Hygiene (LMPH), University of Antwerp, 2610 Wilrijk, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3530-0567","authenticated-orcid":false,"given":"Dorien","family":"Mabille","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, Parasitology and Hygiene (LMPH), University of Antwerp, 2610 Wilrijk, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1714-7549","authenticated-orcid":false,"given":"Roxanne","family":"Mols","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, Parasitology and Hygiene (LMPH), University of Antwerp, 2610 Wilrijk, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6396-0963","authenticated-orcid":false,"given":"Mathieu","family":"Claes","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, Parasitology and Hygiene (LMPH), University of Antwerp, 2610 Wilrijk, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2955-5096","authenticated-orcid":false,"given":"Kayhan","family":"Ilbeigi","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, Parasitology and Hygiene (LMPH), University of Antwerp, 2610 Wilrijk, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rokaya","family":"Ahmad","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, Parasitology and Hygiene (LMPH), University of Antwerp, 2610 Wilrijk, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura","family":"Dirkx","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, Parasitology and Hygiene (LMPH), University of Antwerp, 2610 Wilrijk, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4652-2387","authenticated-orcid":false,"given":"Sara I.","family":"Van Acker","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, Parasitology and Hygiene (LMPH), University of Antwerp, 2610 Wilrijk, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4870-3202","authenticated-orcid":false,"given":"Guy","family":"Caljon","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, Parasitology and Hygiene (LMPH), University of Antwerp, 2610 Wilrijk, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"749","DOI":"10.1016\/S0006-2952(99)00096-9","article-title":"Reporter gene technology: The future looks bright","volume":"58","author":"Naylor","year":"1999","journal-title":"Biochem. Pharmacol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/j.pt.2019.01.002","article-title":"Toolbox for in vivo imaging of host-parasite interactions at multiple scales","volume":"35","author":"Spadin","year":"2019","journal-title":"Trends Parasitol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"6048","DOI":"10.1039\/C6CS00296J","article-title":"1001 lights: Luciferins, luciferases, their mechanisms of action and applications in chemical analysis, biology and medicine","volume":"45","author":"Kaskova","year":"2016","journal-title":"Chem. Soc. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1039\/b719181b","article-title":"Firefly luminescence: A historical perspective and recent developments","volume":"7","author":"Fraga","year":"2008","journal-title":"Photochem. Photobiol. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1007\/s00018-009-0170-8","article-title":"Firefly luciferase: An adenylate-forming enzyme for multicatalytic functions","volume":"67","author":"Inouye","year":"2010","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1038\/nm0103-123","article-title":"Shedding light onto live molecular targets","volume":"9","author":"Weissleder","year":"2003","journal-title":"Nat. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1088\/0031-9155\/25\/4\/008","article-title":"A theoretical and experimental study of light absorption and scattering by In Vivo skin","volume":"25","author":"Dawson","year":"1980","journal-title":"Phys. Med. Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2275","DOI":"10.1007\/s00259-008-0870-6","article-title":"Uptake kinetics and biodistribution of 14C-D-luciferin\u2014A radiolabeled substrate for the firefly luciferase catalyzed bioluminescence reaction: Impact on bioluminescence based reporter gene imaging","volume":"35","author":"Berger","year":"2008","journal-title":"Eur. J. Nucl. Med. Mol. Imaging"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1002\/anie.201205151","article-title":"Rational design and development of near-infrared-emitting firefly luciferins available In Vivo","volume":"52","author":"Kojima","year":"2013","journal-title":"Angew. Chem. Int. Ed. Engl."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"11103","DOI":"10.1021\/bi060475o","article-title":"Luciferin derivatives for enhanced In Vitro and In Vivo bioluminescence assays","volume":"45","author":"Shinde","year":"2006","journal-title":"Biochemistry"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3350","DOI":"10.1002\/anie.201105653","article-title":"A Selenium analogue of firefly D-luciferin with red-shifted bioluminescence emission","volume":"51","author":"Conley","year":"2012","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1649","DOI":"10.1016\/j.chembiol.2011.09.019","article-title":"Identification of mutant firefly luciferases that efficiently utilize aminoluciferins","volume":"18","author":"Harwood","year":"2011","journal-title":"Chem. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"016004","DOI":"10.1117\/1.JBO.17.1.016004","article-title":"Comparison of red-shifted firefly luciferase Ppy RE9 and conventional Luc2 as bioluminescence imaging reporter genes for In Vivo imaging of stem cells","volume":"17","author":"Liang","year":"2012","journal-title":"J. Biomed. Opt."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/j.cbpa.2014.07.003","article-title":"Beyond D-luciferin: Expanding the scope of bioluminescence imaging In Vivo","volume":"21","author":"Adams","year":"2014","journal-title":"Curr. Opin. Chem. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1038\/nmeth.2839","article-title":"A synthetic luciferin improves bioluminescence imaging in live mice","volume":"11","author":"Evans","year":"2014","journal-title":"Nat. Methods"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3847","DOI":"10.1016\/j.tet.2013.03.050","article-title":"Development of simple firefly luciferin analogs emitting blue, green, red, and near-infrared biological window light","volume":"69","author":"Iwano","year":"2013","journal-title":"Tetrahedron"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"11856","DOI":"10.1038\/ncomms11856","article-title":"A luciferin analogue generating near-infrared bioluminescence achieves highly sensitive deep-tissue imaging","volume":"7","author":"Kuchimaru","year":"2016","journal-title":"Nat. Commun."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1111\/j.1462-5822.2004.00468.x","article-title":"Bioluminescent Leishmania expressing luciferase for rapid and high throughput screening of drugs acting on amastigote-harbouring macrophages and for quantitative real-time monitoring of parasitism features in living mice","volume":"7","author":"Lang","year":"2005","journal-title":"Cell. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Michel, G., Ferrua, B., Lang, T., Maddugoda, M.P., Munro, P., Pomares, C., Lemichez, E., and Marty, P. (2011). Luciferase-expressing Leishmania infantum allows the monitoring of amastigote population size, In Vivo, Ex Vivo and In Vitro. PLoS Negl. Trop. Dis., 5.","DOI":"10.1371\/journal.pntd.0001323"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Melo, G.D., Goyard, S., Lecoeur, H., Rouault, E., Pescher, P., Fiette, L., Boissonnas, A., Minoprio, P., and Lang, T. (2017). New insights into experimental visceral leishmaniasis: Real-time In Vivo imaging of Leishmania donovani virulence. PLoS Negl. Trop. Dis., 11.","DOI":"10.1371\/journal.pntd.0005924"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1007\/978-1-4939-9210-2_16","article-title":"Quantification of Leishmania parasites in murine models of visceral infection","volume":"1971","author":"Tavares","year":"2019","journal-title":"Methods Mol. Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1016\/j.parint.2013.07.011","article-title":"In Vivo imaging of trypanosomes for a better assessment of host\u2013parasite relationships and drug efficacy","volume":"63","author":"Goyard","year":"2014","journal-title":"Parasitol. Int."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Bult\u00e9, D., Van Bockstal, L., Dirkx, L., Van den Kerkhof, M., De Trez, C., Timmermans, J.P., Hendrickx, S., Maes, L., and Caljon, G. (2021). Miltefosine enhances infectivity of a miltefosine-resistant Leishmania infantum strain by attenuating its innate immune recognition. PLoS Negl. Trop. Dis., 15.","DOI":"10.1101\/2021.02.22.432409"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"18989","DOI":"10.1038\/s41598-019-55474-3","article-title":"Murine infection with bioluminescent Leishmania infantum axenic amastigotes applied to drug discovery","volume":"9","author":"Tavares","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"\u00c1lvarez-Velilla, R., Guti\u00e9rrez-Corbo, M.d.C., Punz\u00f3n, C., P\u00e9rez-Pertejo, M.Y., Bala\u00f1a-Fouce, R., Fresno, M., and Reguera, R.M. (2019). A chronic bioluminescent model of experimental visceral leishmaniasis for accelerating drug discovery. PLoS Negl. Trop. Dis., 13.","DOI":"10.1371\/journal.pntd.0007133"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Lecoeur, H., Buffet, P., Morizot, G., Goyard, S., Guigon, G., Milon, G., and Lang, T. (2007). Optimization of topical therapy for Leishmania major localized cutaneous leishmaniasis using a reliable C57BL\/6 model. PLoS Negl. Trop. Dis., 1.","DOI":"10.1371\/journal.pntd.0000034"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Costa, D.M., Cec\u00edlio, P., Santar\u00e9m, N., Cordeiro-da-Silva, A., and Tavares, J. (2018). Whole-mouse In Vivo bioluminescence imaging applied to drug screening against Leishmania infantum: A reliable method to evaluate efficacy and optimize treatment regimens. bioRxiv.","DOI":"10.1101\/326355"},{"key":"ref_28","unstructured":"Michels, P.A.M., Ginger, M.L., and Zilberstein, D. (2020). In Vivo bioluminescence imaging to assess compound efficacy against Trypanosoma brucei. Trypanosomatids: Methods and Protocols, Springer."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1177\/1087057114552623","article-title":"A new experimental model for assessing drug efficacy against Trypanosoma cruzi infection based on highly sensitive In Vivo imaging","volume":"20","author":"Lewis","year":"2015","journal-title":"J. Biomol. Screen."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"McLatchie, A.P., Burrell-Saward, H., Myburgh, E., Lewis, M.D., Ward, T.H., Mottram, J.C., Croft, S.L., Kelly, J.M., and Taylor, M.C. (2013). Highly sensitive In Vivo imaging of Trypanosoma brucei expressing \u201cred-shifted\u201d luciferase. PLoS Negl. Trop. Dis., 7.","DOI":"10.1371\/journal.pntd.0002571"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1093\/jac\/dky450","article-title":"Miltefosine enhances the fitness of a non-virulent drug-resistant Leishmania infantum strain","volume":"74","author":"Eberhardt","year":"2019","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"871","DOI":"10.1021\/acsinfecdis.0c00864","article-title":"Bioluminescent imaging identifies thymus, as overlooked colonized organ, in a chronic model of Leishmania donovani mouse visceral leishmaniasis","volume":"7","author":"Iborra","year":"2021","journal-title":"ACS Infect. Dis."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Claes, F., Vodnala, S.K., van Reet, N., Boucher, N., Lunden-Miguel, H., Baltz, T., Goddeeris, B.M., B\u00fcscher, P., and Rottenberg, M.E. (2009). Bioluminescent imaging of Trypanosoma brucei shows preferential testis dissemination which may hamper drug efficacy in sleeping sickness. PLoS Negl. Trop. Dis., 3.","DOI":"10.1371\/journal.pntd.0000486"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"626","DOI":"10.1038\/s42003-022-03591-7","article-title":"Long-term hematopoietic stem cells as a parasite niche during treatment failure in visceral leishmaniasis","volume":"5","author":"Dirkx","year":"2022","journal-title":"Commun. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"105444","DOI":"10.1016\/j.actatropica.2020.105444","article-title":"Evaluation of NanoLuc, RedLuc and Luc2 as bioluminescent reporters in a cutaneous leishmaniasis model","volume":"206","author":"Agostino","year":"2020","journal-title":"Acta Trop."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Van Reet, N., Van de Vyver, H., Pyana, P.P., Van der Linden, A.M., and B\u00fcscher, P. (2014). A panel of Trypanosoma brucei strains tagged with blue and red-shifted luciferases for bioluminescent imaging in murine infection models. PLoS Negl. Trop. Dis., 8.","DOI":"10.1371\/journal.pntd.0003054"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Branchini, B.R., Fontaine, D.M., Kohrt, D., Huta, B.P., Racela, A.R., Fort, B.R., Southworth, T.L., and Roda, A. (2022). Systematic comparison of beetle luciferase-luciferin pairs as sources of near-infrared light for In Vitro and In Vivo applications. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23052451"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"852","DOI":"10.1038\/s41592-020-0889-6","article-title":"Novel NanoLuc substrates enable bright two-population bioluminescence imaging in animals","volume":"17","author":"Su","year":"2020","journal-title":"Nat. Methods"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Nakayama, J., Saito, R., Hayashi, Y., Kitada, N., Tamaki, S., Han, Y., Semba, K., and Maki, S.A. (2020). High sensitivity In Vivo imaging of cancer metastasis using a near-infrared luciferin analogue seMpai. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21217896"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"11203","DOI":"10.1038\/s41598-018-29527-y","article-title":"Neutrophils enhance early Trypanosoma brucei infection onset","volume":"8","author":"Caljon","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"S1","DOI":"10.1111\/j.1948-7134.2011.00106.x","article-title":"Establishment and maintenance of sand fly colonies","volume":"36","author":"Volf","year":"2011","journal-title":"J. Vector Ecol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"951","DOI":"10.1093\/jac\/dkz517","article-title":"Transmission potential of paromomycin-resistant Leishmania infantum and Leishmania donovani","volume":"75","author":"Hendrickx","year":"2019","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1186\/s13071-020-3972-z","article-title":"Phenotypic adaptations of Leishmania donovani to recurrent miltefosine exposure and impact on sand fly infection","volume":"13","author":"Hendrickx","year":"2020","journal-title":"Parasites Vectors"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.ijpddr.2020.04.004","article-title":"Impact of clinically acquired miltefosine resistance by Leishmania infantum on mouse and sand fly infection","volume":"13","author":"Hendrickx","year":"2020","journal-title":"Int. J. Parasitol. Drugs Drug Resist."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"3601","DOI":"10.1007\/s00436-018-6059-4","article-title":"Impact of primary mouse macrophage cell types on Leishmania infection and In Vitro drug susceptibility","volume":"117","author":"Gielis","year":"2018","journal-title":"Parasitol. Res."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"3524","DOI":"10.1128\/AAC.01781-15","article-title":"A Replicative In Vitro assay for drug discovery against Leishmania donovani","volume":"60","author":"Tegazzini","year":"2016","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"762","DOI":"10.1152\/physiolgenomics.00055.2016","article-title":"A synthetic luciferin improves In Vivo bioluminescence imaging of gene expression in cardiovascular brain regions","volume":"48","author":"Simonyan","year":"2016","journal-title":"Physiol. Genom."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1523","DOI":"10.1007\/s11307-020-01523-7","article-title":"Evaluating brightness and spectral properties of click beetle and firefly luciferases using luciferin analogues: Identification of preferred pairings of luciferase and substrate for In Vivo bioluminescence imaging","volume":"22","author":"Zambito","year":"2020","journal-title":"Mol. Imaging Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/24\/16074\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:43:00Z","timestamp":1760146980000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/24\/16074"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,16]]},"references-count":48,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["ijms232416074"],"URL":"https:\/\/doi.org\/10.3390\/ijms232416074","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,16]]}}}