{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,2]],"date-time":"2025-11-02T04:53:37Z","timestamp":1762059217959,"version":"build-2065373602"},"reference-count":45,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2022,7,23]],"date-time":"2022-07-23T00:00:00Z","timestamp":1658534400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Science and Technology, Executive Yuan, Taiwan","award":["MOST-111-2811-B-195-001","MOST-111-2811-B-195-002","MOST-111-2314-B-195-017","MOST-110-2314-B-195-010-MY3","MOST-110-2314-B-195-014","MOST-110-2314-B-195-029","MOST-109-2314-B-195-024","MOST-108-2314-B-195-012","MOST-108-2314-B-195-014","MMH-E-111-13","MMH-E-110-16","MMH-E-109-16","MMH-E-108-16","MMC-RD-109-1B-06","MMC-RD-110-1C-P004"],"award-info":[{"award-number":["MOST-111-2811-B-195-001","MOST-111-2811-B-195-002","MOST-111-2314-B-195-017","MOST-110-2314-B-195-010-MY3","MOST-110-2314-B-195-014","MOST-110-2314-B-195-029","MOST-109-2314-B-195-024","MOST-108-2314-B-195-012","MOST-108-2314-B-195-014","MMH-E-111-13","MMH-E-110-16","MMH-E-109-16","MMH-E-108-16","MMC-RD-109-1B-06","MMC-RD-110-1C-P004"]}]},{"name":"MacKay Memorial Hospital","award":["MOST-111-2811-B-195-001","MOST-111-2811-B-195-002","MOST-111-2314-B-195-017","MOST-110-2314-B-195-010-MY3","MOST-110-2314-B-195-014","MOST-110-2314-B-195-029","MOST-109-2314-B-195-024","MOST-108-2314-B-195-012","MOST-108-2314-B-195-014","MMH-E-111-13","MMH-E-110-16","MMH-E-109-16","MMH-E-108-16","MMC-RD-109-1B-06","MMC-RD-110-1C-P004"],"award-info":[{"award-number":["MOST-111-2811-B-195-001","MOST-111-2811-B-195-002","MOST-111-2314-B-195-017","MOST-110-2314-B-195-010-MY3","MOST-110-2314-B-195-014","MOST-110-2314-B-195-029","MOST-109-2314-B-195-024","MOST-108-2314-B-195-012","MOST-108-2314-B-195-014","MMH-E-111-13","MMH-E-110-16","MMH-E-109-16","MMH-E-108-16","MMC-RD-109-1B-06","MMC-RD-110-1C-P004"]}]},{"name":"MacKay Medical College","award":["MOST-111-2811-B-195-001","MOST-111-2811-B-195-002","MOST-111-2314-B-195-017","MOST-110-2314-B-195-010-MY3","MOST-110-2314-B-195-014","MOST-110-2314-B-195-029","MOST-109-2314-B-195-024","MOST-108-2314-B-195-012","MOST-108-2314-B-195-014","MMH-E-111-13","MMH-E-110-16","MMH-E-109-16","MMH-E-108-16","MMC-RD-109-1B-06","MMC-RD-110-1C-P004"],"award-info":[{"award-number":["MOST-111-2811-B-195-001","MOST-111-2811-B-195-002","MOST-111-2314-B-195-017","MOST-110-2314-B-195-010-MY3","MOST-110-2314-B-195-014","MOST-110-2314-B-195-029","MOST-109-2314-B-195-024","MOST-108-2314-B-195-012","MOST-108-2314-B-195-014","MMH-E-111-13","MMH-E-110-16","MMH-E-109-16","MMH-E-108-16","MMC-RD-109-1B-06","MMC-RD-110-1C-P004"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JPM"],"abstract":"<jats:p>Mucopolysaccharidosis type I (MPS I) is an inherited autosomal recessive disease resulting from mutation of the \u03b1-l-Iduronidase (IDUA) gene. New unknown mutated nucleotides of idua have increasingly been discovered in newborn screening, and remain to be elucidated. In this study, we found that the z-Idua enzymatic activity of zebrafish idua-knockdown embryos was reduced, resulting in the accumulation of undegradable metabolite of heparin sulfate, as well as increased mortality and defective phenotypes similar to some symptoms of human MPS I. After microinjecting mutated z-idua-L346R, -T364M, -E398-deleted, and -E540-frameshifted mRNAs, corresponding to mutated human IDUA associated with MPS I, into zebrafish embryos, no increase in z-Idua enzymatic activity, except of z-idua-E540-frameshift-injected embryos, was noted compared with endogenous z-Idua of untreated embryos. Defective phenotypes were observed in the z-idua-L346R-injected embryos, suggesting that failed enzymatic activity of mutated z-Idua-L346R might have a dominant negative effect on endogenous z-Idua function. However, defective phenotypes were not observed in the z-idua-E540-frameshifted-mRNA-injected embryos, which provided partial enzymatic activity. Based on these results, we suggest that the z-Idua enzyme activity assay combined with phenotypic observation of mutated-idua-injected zebrafish embryos could serve as an alternative platform for a preliminary assessment of mutated idua not yet characterized for their role in MPS I.<\/jats:p>","DOI":"10.3390\/jpm12081199","type":"journal-article","created":{"date-parts":[[2022,7,24]],"date-time":"2022-07-24T22:49:02Z","timestamp":1658702942000},"page":"1199","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Quantification of Idua Enzymatic Activity Combined with Observation of Phenotypic Change in Zebrafish Embryos Provide a Preliminary Assessment of Mutated idua Correlated with Mucopolysaccharidosis Type I"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9570-0218","authenticated-orcid":false,"given":"Cheng-Yung","family":"Lin","sequence":"first","affiliation":[{"name":"Institute of Biomedical Sciences, MacKay Medical College, New Taipei City 25245, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9619-0990","authenticated-orcid":false,"given":"Hsiang-Yu","family":"Lin","sequence":"additional","affiliation":[{"name":"Institute of Biomedical Sciences, MacKay Medical College, New Taipei City 25245, Taiwan"},{"name":"Department of Medicine, MacKay Medical College, New Taipei City 25245, Taiwan"},{"name":"Department of Pediatrics, MacKay Memorial Hospital, Taipei 10449, Taiwan"},{"name":"Department of Medical Research, MacKay Memorial Hospital, New Taipei City 25160, Taiwan"},{"name":"MacKay Junior College of Medicine, Nursing and Management, Taipei 11260, Taiwan"},{"name":"Department of Medical Research, China Medical University Hospital, China Medical University, Taichung 40402, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9776-2092","authenticated-orcid":false,"given":"Chih-Kuang","family":"Chuang","sequence":"additional","affiliation":[{"name":"Department of Medical Research, MacKay Memorial Hospital, New Taipei City 25160, Taiwan"},{"name":"School of Medicine, Fu Jen Catholic University, New Taipei City 242062, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Po-Hsiang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Medical Research, MacKay Memorial Hospital, New Taipei City 25160, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuan-Rong","family":"Tu","sequence":"additional","affiliation":[{"name":"Department of Medical Research, MacKay Memorial Hospital, New Taipei City 25160, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuan-Pei","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Medicine, MacKay Medical College, New Taipei City 25245, Taiwan"},{"name":"Department of Pediatrics, MacKay Memorial Hospital, Taipei 10449, Taiwan"},{"name":"Department of Medical Research, MacKay Memorial Hospital, New Taipei City 25160, Taiwan"},{"name":"Department of Infant and Child Care, National Taipei University of Nursing and Health Sciences, Taipei 11219, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huai-Jen","family":"Tsai","sequence":"additional","affiliation":[{"name":"School of Medicine, Fu Jen Catholic University, New Taipei City 242062, Taiwan"},{"name":"Department of Life Science, Fu Jen Catholic University, New Taipei City 242062, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1186\/s13023-015-0337-3","article-title":"Decreased performance in IDUA knockout mouse mimic limitationsofjointfunction and locomotion in patients with Hurler syndrome","volume":"10","author":"Kim","year":"2015","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1186\/s13023-017-0662-9","article-title":"Open issues in Mucopolysaccharidosis type I-Hurler","volume":"12","author":"Parini","year":"2017","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"360","DOI":"10.1016\/0021-9681(56)90138-2","article-title":"Heritable disorders of connective tissue","volume":"3","author":"Mckusick","year":"1956","journal-title":"J. Chronic Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1542\/peds.2008-0416","article-title":"Mucopolysaccharidosis I: Management and treatment guidelines","volume":"123","author":"Muenzer","year":"2009","journal-title":"Pediatrics"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"750","DOI":"10.1007\/s10545-007-0586-y","article-title":"Cardiac abnormalities in adults with the attenuated form of mucopolysaccharidosis type I","volume":"30","author":"Soliman","year":"2007","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"924","DOI":"10.1590\/S1415-47572012000600006","article-title":"Mucopolysacccharidoses: From understanding to treatment, a century of discoveries","volume":"35","author":"Giugliani","year":"2012","journal-title":"Genet. Mol. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1186\/s13023-017-0733-y","article-title":"Failure to shorten the diagnostic delay in two ultra-orphan diseases (mucopolysaccharidosis types I and III): Potential causes and implications","volume":"13","author":"Kuiper","year":"2018","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Hampe, C.S., Wesley, J., Lund, T.C., Orchard, P.J., Polgreen, L.E., Eisengart, J.B., McLoon, L.K., Cureoglu, S., Schachern, P., and McIvor, R.S. (2021). Mucopolysaccharidosis Type I: Current Treatments, Limitations, and Prospects for Improvement. Biomolecules, 11.","DOI":"10.3390\/biom11020189"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1186\/s13052-018-0552-3","article-title":"Newborn screening in mucopolysaccharidoses","volume":"44","author":"Donati","year":"2018","journal-title":"Ital. J. Pediatr."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Clarke, L.A., Dickson, P., Ellinwood, N.M., and Klein, T.L. (2020). Newborn Screening for Mucopolysaccharidosis I: Moving Forward Learning from Experience. Int. J. Neonatal Screen., 6.","DOI":"10.3390\/ijns6040091"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Chuang, C.K., Lee, C.L., Tu, R.Y., Lo, Y.T., Sisca, F., Chang, Y.H., Liu, M.Y., Liu, H.Y., Chen, H.J., and Kao, S.M. (2021). Nationwide Newborn Screening Program for Mucopolysaccharidoses in Taiwan and an Update of the \u201cGold Standard\u201d Criteria Required to Make a Confirmatory Diagnosis. Diagnostics, 11.","DOI":"10.3390\/diagnostics11091583"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1186\/s13023-018-0816-4","article-title":"Status of newborn screening and follow up investigations for Mucopolysaccharidoses I and II in Taiwan","volume":"13","author":"Chuang","year":"2018","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1111\/cge.13583","article-title":"Genotype-phenotype relationships in mucopolysaccharidosis type I (MPS I): Insights from the International MPS I Registry","volume":"96","author":"Clarke","year":"2019","journal-title":"Clin. Genet."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1186\/s12929-016-0236-5","article-title":"Zebrafish and Medaka: New model organisms for modern biomedical research","volume":"23","author":"Lin","year":"2016","journal-title":"J. Biomed. Sci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1016\/j.cbpa.2014.10.025","article-title":"15 years of zebrafish chemical screening","volume":"24","author":"Rennekamp","year":"2015","journal-title":"Curr. Opin. Chem. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Lee, H.C., Lin, C.Y., and Tsai, H.J. (2021). Zebrafish, an In Vivo Platform to Screen Drugs and Proteins for Biomedical Use. Pharmaceuticals, 14.","DOI":"10.3390\/ph14060500"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1038\/s41572-018-0025-4","article-title":"Lysosomal storage diseases","volume":"4","author":"Platt","year":"2018","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"82","DOI":"10.3389\/fmolb.2020.00082","article-title":"Modeling Lysosomal Storage Diseases in the Zebrafish","volume":"7","author":"Zhang","year":"2020","journal-title":"Front. Mol. Biosci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.matbio.2009.09.001","article-title":"A novel functional role of iduronate-2-sulfatase in zebrafish early development","volume":"29","author":"Moro","year":"2010","journal-title":"Matrix Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1643","DOI":"10.1093\/hmg\/ddx069","article-title":"Perturbations in cell signaling elicit early cardiac defects in mucopolysaccharidosis type II","volume":"26","author":"Costa","year":"2017","journal-title":"Hum. Mol. Genet."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1186\/s13023-020-01598-z","article-title":"Survival and diagnostic age of 175 Taiwanese patients with mucopolysaccharidoses (1985\u20132019)","volume":"15","author":"Lin","year":"2020","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2262","DOI":"10.1093\/hmg\/ddy131","article-title":"FGF signaling deregulation is associated with early developmental skeletal defects in animal models for mucopolysaccharidosis type II (MPSII)","volume":"27","author":"Bellesso","year":"2018","journal-title":"Hum. Mol. Genet."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Lin, C.Y., Lin, H.Y., Chuang, C.K., Zhang, P.H., Tu, R.Y., Lin, S.P., and Tsai, H.J. (2020). Effect of Mutated ids Overexpression on IDS Enzyme Activity and Developmental Phenotypes in Zebrafish Embryos: A Valuable Index for Assessing Critical Point-Mutations Associated with Mucopolysaccharidosis Type II Occurrence in Humans. Diagnostics, 10.","DOI":"10.3390\/diagnostics10100854"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Douek, A.M., Amiri Khabooshan, M., Henry, J., Stamatis, S.A., Kreuder, F., Ramm, G., Anko, M.L., Wlodkowic, D., and Kaslin, J. (2021). An Engineered sgsh Mutant Zebrafish Recapitulates Molecular and Behavioural Pathobiology of Sanfilippo Syndrome A\/MPS IIIA. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22115948"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"960","DOI":"10.1002\/ajmg.a.32781","article-title":"Incidence of the mucopolysaccharidoses in Taiwan, 1984\u20132004","volume":"149A","author":"Lin","year":"2009","journal-title":"Am. J. Med. Genet. A"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1007\/s004390050641","article-title":"Incidence of the mucopolysaccharidoses in Northern Ireland","volume":"101","author":"Nelson","year":"1997","journal-title":"Hum. Genet."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1011","DOI":"10.1007\/s10545-005-0112-z","article-title":"Cumulative incidence rates of the mucopolysaccharidoses in Germany","volume":"28","author":"Baehner","year":"2005","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1007\/s13353-014-0262-5","article-title":"Prevalence rates of mucopolysaccharidoses in Poland","volume":"56","author":"Jurecka","year":"2015","journal-title":"J. Appl. Genet."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"79","DOI":"10.5582\/irdr.2014.01013","article-title":"An overview of Korean patients with mucopolysaccharidosis and collaboration through the Asia Pacific MPS Network","volume":"3","author":"Cho","year":"2014","journal-title":"Intractable Rare Dis. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1038\/s41436-018-0129-y","article-title":"The New York pilot newborn screening program for lysosomal storage diseases: Report of the First 65,000 Infants","volume":"21","author":"Wasserstein","year":"2019","journal-title":"Genet. Med."},{"key":"ref_31","unstructured":"Westerfield, M. (1995). The Zebrafish Book, University of Oregon Press. [3rd ed.]."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1186\/s13023-014-0135-3","article-title":"A modified liquid chromatography\/tandem mass spectrometry method for predominant disaccharide units of urinary glycosaminoglycans in patients with mucopolysaccharidoses","volume":"9","author":"Chuang","year":"2014","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"170009","DOI":"10.1098\/rsob.170009","article-title":"microRNA-206 modulates an Rtn4a\/Cxcr4a\/Thbs3a axis in newly forming somites to maintain and stabilize the somite boundary formation of zebrafish embryos","volume":"7","author":"Lin","year":"2017","journal-title":"Open Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2098","DOI":"10.1093\/clinchem\/47.12.2098","article-title":"Hurler-like phenotype- enzymatic diagnosis in dried blood spots on filter paper","volume":"47","author":"Chamoles","year":"2001","journal-title":"Clin. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1186\/1750-1172-8-147","article-title":"A pilot newborn screening program for Mucopolysaccharidosis type I in Taiwan","volume":"8","author":"Lin","year":"2013","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1089\/zeb.2013.0903","article-title":"Normal function of Myf5 during gastrulation is required for pharyngeal arch cartilage development in zebrafish embryos","volume":"10","author":"Lin","year":"2013","journal-title":"Zebrafish"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1007\/s10126-018-9857-x","article-title":"Conditional Overexpression of rtn4al in Muscle of Adult Zebrafish Displays Defects Similar to Human Amyotrophic Lateral Sclerosis","volume":"21","author":"Lin","year":"2019","journal-title":"Mar. Biotechnol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1038\/nchembio.1357","article-title":"Insights into mucopolysaccharidosis I from the structure and action of alpha-L-iduronidase","volume":"9","author":"Bie","year":"2013","journal-title":"Nat. Chem. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"14628","DOI":"10.1073\/pnas.1306939110","article-title":"Human alpha-L-iduronidase uses its own N-glycan as a substrate-binding and catalytic module","volume":"110","author":"Maita","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.pedneo.2018.01.004","article-title":"Awareness of attenuated mucopolysaccharidoses in a pediatric orthopedic clinic","volume":"60","author":"Lin","year":"2019","journal-title":"Pediatr. Neonatol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1111\/j.1399-0004.2004.00374.x","article-title":"Mutational analysis of the alpha-L-iduronidase gene in 10 unrelated Korean type I mucopolysaccharidosis patients: Identification of four novel mutations","volume":"66","author":"Lee","year":"2004","journal-title":"Clin. Genet."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1034\/j.1399-0004.1999.560109.x","article-title":"Mucopolysaccharidosis type I: Characterization of novel mutations affecting alpha-L-iduronidase activity","volume":"56","author":"Lin","year":"1999","journal-title":"Clin. Genet."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Habicher, J., Haitina, T., Eriksson, I., Holmborn, K., Dierker, T., Ahlberg, P.E., and Ledin, J. (2015). Chondroitin\/dermatan sulfate modification enzymes in zebrafish development. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0121957"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"964","DOI":"10.1002\/dvdy.23981","article-title":"Expression of chondroitin\/dermatan sulfate glycosyltransferases during early zebrafish development","volume":"242","author":"Holmborn","year":"2013","journal-title":"Dev. Dyn."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"194106","DOI":"10.1088\/0953-8984\/23\/19\/194106","article-title":"The Donnan equilibrium: I. On the thermodynamic foundation of the Donnan equation of state","volume":"23","author":"Philipse","year":"2011","journal-title":"J. Phys. Condens. Matter"}],"container-title":["Journal of Personalized Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4426\/12\/8\/1199\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:55:18Z","timestamp":1760140518000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4426\/12\/8\/1199"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,23]]},"references-count":45,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2022,8]]}},"alternative-id":["jpm12081199"],"URL":"https:\/\/doi.org\/10.3390\/jpm12081199","relation":{},"ISSN":["2075-4426"],"issn-type":[{"type":"electronic","value":"2075-4426"}],"subject":[],"published":{"date-parts":[[2022,7,23]]}}}