{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,8]],"date-time":"2025-11-08T22:52:13Z","timestamp":1762642333742,"version":"build-2065373602"},"reference-count":60,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2020,3,17]],"date-time":"2020-03-17T00:00:00Z","timestamp":1584403200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Finnish Eye and Tissue Bank Foundation","award":["-"],"award-info":[{"award-number":["-"]}]},{"DOI":"10.13039\/501100008413","name":"Instrumentarium Science Foundation","doi-asserted-by":"publisher","award":["-"],"award-info":[{"award-number":["-"]}],"id":[{"id":"10.13039\/501100008413","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The induction of heat shock response in the macula has been proposed as a useful therapeutic strategy for retinal neurodegenerative diseases by promoting proteostasis and enhancing protective chaperone mechanisms. We applied transpupillary 1064 nm long-duration laser heating to the mouse (C57Bl\/6J) fundus to examine the heat shock response in vivo. The intensity and spatial distribution of heat shock protein (HSP) 70 expression along with the concomitant probability for damage were measured 24 h after laser irradiation in the mouse retinal pigment epithelium (RPE) as a function of laser power. Our results show that the range of heating powers for producing heat shock response while avoiding damage in the mouse RPE is narrow. At powers of 64 and 70 mW, HSP70 immunostaining indicates 90 and 100% probability for clearly elevated HSP expression while the corresponding probability for damage is 20 and 33%, respectively. Tunel staining identified the apoptotic regions, and the estimated 50% damaging threshold probability for the heating (ED50) was ~72 mW. The staining with Bestrophin1 (BEST1) demonstrated RPE cell atrophy with the most intense powers. Consequently, fundus heating with a long-duration laser provides an approachable method to develop heat shock-based therapies for the RPE of retinal disease model mice.<\/jats:p>","DOI":"10.3390\/ijms21062063","type":"journal-article","created":{"date-parts":[[2020,3,18]],"date-time":"2020-03-18T08:13:27Z","timestamp":1584519207000},"page":"2063","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Induction of Heat Shock Protein 70 in Mouse RPE as an In Vivo Model of Transpupillary Thermal Stimulation"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1652-9307","authenticated-orcid":false,"given":"Mooud","family":"Amirkavei","sequence":"first","affiliation":[{"name":"Department of Neuroscience and Biomedical Engineering, School of Science, Aalto University, 00076 Espoo, Finland"},{"name":"Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, 17177 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6054-8380","authenticated-orcid":false,"given":"Marja","family":"Pitk\u00e4nen","sequence":"additional","affiliation":[{"name":"Department of Neuroscience and Biomedical Engineering, School of Science, Aalto University, 00076 Espoo, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ossi","family":"Kaikkonen","sequence":"additional","affiliation":[{"name":"Department of Neuroscience and Biomedical Engineering, School of Science, Aalto University, 00076 Espoo, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kai","family":"Kaarniranta","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Institute of Clinical Medicine, University of Eastern Finland and Kuopio University Hospital, 70210 Kuopio, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2926-2376","authenticated-orcid":false,"given":"Helder","family":"Andr\u00e9","sequence":"additional","affiliation":[{"name":"Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, 17177 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ari","family":"Koskelainen","sequence":"additional","affiliation":[{"name":"Department of Neuroscience and Biomedical Engineering, School of Science, Aalto University, 00076 Espoo, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,3,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"670","DOI":"10.1007\/s00018-004-4464-6","article-title":"Hsp70 chaperones: Cellular functions and molecular mechanism","volume":"62","author":"Mayer","year":"2005","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_2","first-page":"74","article-title":"Antiapoptotic and anti-inflammatory mechanisms of heat-shock protein protection","volume":"1053","author":"Yenari","year":"2005","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1111\/j.1582-4934.2008.00273.x","article-title":"Heat shock proteins: Essential proteins for apoptosis regulation: Apoptosis Review Series","volume":"12","author":"Lanneau","year":"2008","journal-title":"J. Cell. Mol. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/j.molcel.2010.10.006","article-title":"The Heat Shock Response: Life on the Verge of Death","volume":"40","author":"Richter","year":"2010","journal-title":"Mol. Cell"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Penke, B., Bog\u00e1r, F., Crul, T., S\u00e1ntha, M., T\u00f3th, M.E., and V\u00edgh, L. (2018). Heat shock proteins and autophagy pathways in neuroprotection: From molecular bases to pharmacological interventions. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19010325"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1016\/S1474-4422(16)00099-5","article-title":"Heat shock proteins as potential targets for protective strategies in neurodegeneration","volume":"15","author":"Kampinga","year":"2016","journal-title":"Lancet Neurol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1016\/S0039-6257(03)00030-4","article-title":"Age-related macular degeneration: Etiology, pathogenesis, and therapeutic strategies","volume":"48","author":"Ambati","year":"2003","journal-title":"Surv. Ophthalmol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.neuron.2012.06.018","article-title":"Mechanisms of age-related macular degeneration","volume":"75","author":"Ambati","year":"2012","journal-title":"Neuron"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Hyttinen, J.M.T., Kannan, R., Felszeghy, S., Niittykoski, M., Salminen, A., and Kaarniranta, K. (2019). The regulation of NFE2L2 (NRF2) signalling and epithelial-to-mesenchymal transition in age-related macular degeneration pathology. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20225800"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1079","DOI":"10.1016\/j.ophtha.2013.11.023","article-title":"Geographic atrophy: Clinical features and potential therapeutic approaches","volume":"121","author":"Holz","year":"2014","journal-title":"Ophthalmology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1016\/j.mam.2012.04.006","article-title":"Retinal pigment epithelium response to oxidant injury in the pathogenesis of early age-related macular degeneration","volume":"33","author":"Mettu","year":"2012","journal-title":"Mol. Aspects Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1038\/nm1709","article-title":"Oxidative damage-induced inflammation initiates age-related macular degeneration","volume":"14","author":"Hollyfield","year":"2008","journal-title":"Nat. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1430","DOI":"10.1172\/JCI71029","article-title":"Recent developments in the treatment of age-related macular degeneration","volume":"124","author":"Holz","year":"2014","journal-title":"J. Clin. Invest."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1111\/j.1755-3768.2011.02179.x","article-title":"Molecular mechanisms of retinal pigment epithelium damage and development of age-related macular degeneration","volume":"90","author":"Kinnunen","year":"2012","journal-title":"Acta Ophthalmol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1007\/s00018-019-03422-9","article-title":"The role of hypoxia-inducible factors in neovascular age-related macular degeneration: a gene therapy perspective","volume":"77","author":"Mammadzada","year":"2020","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1016\/S0197-0186(02)00043-8","article-title":"Chaperones and aging: Role in neurodegeneration and in other civilizational diseases","volume":"41","author":"Csermely","year":"2002","journal-title":"Neurochem. Int."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"30","DOI":"10.3109\/08820538.2015.1114837","article-title":"Subthreshold diode micropulse laser: A review","volume":"31","author":"Brader","year":"2016","journal-title":"Semin. Ophthalmol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2488","DOI":"10.1167\/iovs.15-18981","article-title":"Nondamaging retinal laser therapy: Rationale and applications to the macula","volume":"57","author":"Lavinsky","year":"2016","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1780","DOI":"10.1167\/iovs.10-5917","article-title":"Non-damaging retinal phototherapy: Dynamic range of heat shock protein expression","volume":"52","author":"Sramek","year":"2011","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_20","first-page":"969319","article-title":"Heat shock protein expression as guidance for the therapeutic window of retinal laser therapy","volume":"9693","author":"Wang","year":"2016","journal-title":"Ophthalmic Technol. XXVI"},{"key":"ref_21","first-page":"2976","article-title":"Heat shock protein hyperexpression on chorioretinal layers after transpupillary thermotherapy","volume":"42","author":"Desmettre","year":"2001","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1143","DOI":"10.1007\/s00417-005-0042-3","article-title":"Subthreshold transpupillary thermotherapy of the retina and experimental choroidal neovascularization in a rat model","volume":"244","author":"She","year":"2006","journal-title":"Graefe\u2019s Arch. Clin. Exp. Ophthalmol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"729792","DOI":"10.1155\/2015\/729792","article-title":"Sublethal photothermal stimulation with a micropulse laser induces heat shock protein expression in ARPE-19 Cells","volume":"2015","author":"Inagaki","year":"2015","journal-title":"J. Ophthalmol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.exer.2018.02.013","article-title":"Expression of heat shock protein 70 and cell death kinetics after different thermal impacts on cultured retinal pigment epithelial cells","volume":"170","author":"Kern","year":"2018","journal-title":"Exp. Eye Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"078002","DOI":"10.1117\/1.3600013","article-title":"Pulse duration determines levels of Hsp70 induction in tissues following laser irradiation","volume":"16","author":"Mackanos","year":"2011","journal-title":"J. Biomed. Opt."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1510","DOI":"10.1001\/archopht.122.10.1510","article-title":"Histological effect and protein expression in subthreshold transpupillary thermotherapy in rabbit eyes","volume":"122","author":"Morimura","year":"2004","journal-title":"Arch. Ophthalmol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"8643","DOI":"10.1073\/pnas.90.18.8643","article-title":"Microphotocoagulation: selective effects of repetitive short laser pulses","volume":"90","author":"Roider","year":"1993","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_28","first-page":"337","article-title":"Biochemistry of visual pigment regeneration: The Friedenwald lecture","volume":"41","author":"Saari","year":"2000","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"516","DOI":"10.1016\/j.survophthal.2010.02.005","article-title":"Micropulsed diode laser therapy: evolution and clinical applications","volume":"55","author":"Sivaprasad","year":"2010","journal-title":"Surv. Ophthalmol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1097\/IAE.0b013e3182206f6c","article-title":"Long-term safety, high-resolution imaging, and tissue temperature modeling of subvisible diode micropulse photocoagulation for retinovascular macular edema","volume":"32","author":"Luttrull","year":"2012","journal-title":"Retina"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4722","DOI":"10.1167\/iovs.17-21610","article-title":"Comparison of continuous-wave and micropulse modulation in retinal laser therapy","volume":"58","author":"Wang","year":"2017","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_32","first-page":"2380","article-title":"Heat shock protein 70 expression induced by diode laser irradiation on choroid-retinal endothelial cells in vitro","volume":"18","author":"Du","year":"2012","journal-title":"Mol. Vis."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/0014-4835(86)90026-6","article-title":"Acute reversible cataract induced by xylazine and by ketamine-xylazine anesthesia in rats and mice","volume":"42","author":"Calderone","year":"1986","journal-title":"Exp. Eye Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1006\/exer.2002.2007","article-title":"Causes of cataract development in anesthetized mice","volume":"75","author":"Ridder","year":"2002","journal-title":"Exp. Eye Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3678","DOI":"10.1167\/iovs.04-0436","article-title":"Retinal temperature increase during transpupillary thermotherapy: Effects of pigmentation, subretinal blood, and choroidal blood flow","volume":"45","author":"Ibarra","year":"2004","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1007\/s10633-011-9271-4","article-title":"Effects of common anesthetics on eye movement and electroretinogram","volume":"122","author":"Nair","year":"2011","journal-title":"Doc. Ophthalmol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"034007","DOI":"10.1117\/1.3130282","article-title":"Dynamics of retinal photocoagulation and rupture","volume":"14","author":"Sramek","year":"2009","journal-title":"J. Biomed. Opt."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1109\/TBME.1970.4502736","article-title":"The Thermal-Chemical Damage in Biological Material under Laser Irradiation","volume":"BME-17","author":"Hu","year":"1970","journal-title":"IEEE Trans. Biomed. Eng."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-019-43906-z","article-title":"A lasered mouse model of retinal degeneration displays progressive outer retinal pathology providing insights into early geographic atrophy","volume":"9","author":"Ibbett","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2360","DOI":"10.1007\/s10439-017-1872-y","article-title":"A novel method for mouse retinal temperature determination based on ERG photoresponses","volume":"45","author":"Kaikkonen","year":"2017","journal-title":"Ann. Biomed. Eng."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"107675","DOI":"10.1016\/j.exer.2019.05.015","article-title":"In vivo monitoring of mouse retinal temperature by ERG photoresponses","volume":"187","author":"Kaikkonen","year":"2019","journal-title":"Exp. Eye Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.arr.2009.01.001","article-title":"Heat shock proteins as gatekeepers of proteolytic pathways-Implications for age-related macular degeneration (AMD)","volume":"8","author":"Kaarniranta","year":"2009","journal-title":"Ageing Res. Rev."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1007\/s11095-014-1456-6","article-title":"Oxidative stress protection by exogenous delivery of rhhsp70 chaperone to the retinal pigment epithelium (RPE), a possible therapeutic strategy against RPE degeneration","volume":"32","author":"Subrizi","year":"2015","journal-title":"Pharm. Res."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3616","DOI":"10.1111\/j.1582-4934.2008.00577.x","article-title":"Crosstalk between Hsp70 molecular chaperone, lysosomes and proteasomes in autophagy-mediated proteolysis in human retinal pigment epithelial cells","volume":"13","author":"Hyttinen","year":"2009","journal-title":"J. Cell. Mol. Med."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"935","DOI":"10.1016\/j.cell.2016.07.001","article-title":"UBQLN2 Mediates Autophagy-Independent Protein Aggregate Clearance by the Proteasome","volume":"166","author":"Hjerpe","year":"2016","journal-title":"Cell"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1379\/CSC-265","article-title":"Heat Shock Protein 70 Regulates Cellular Redox Status by Modulating Glutathione Related Enzyme Activities","volume":"preprint","author":"Guo","year":"2005","journal-title":"Cell Stress Chaperones"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1097\/IAE.0000000000000340","article-title":"Nondamaging photothermal therapy for the retina: Initial clinical experience with chronic central serous retinopathy","volume":"35","author":"Lavinsky","year":"2015","journal-title":"Retina"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1136\/bjo.2004.051540","article-title":"Subthreshold diode micropulse photocoagulation for the treatment of clinically significant diabetic macular oedema","volume":"89","author":"Luttrull","year":"2005","journal-title":"Br. J. Ophthalmol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"4314","DOI":"10.1167\/iovs.10-6828","article-title":"Randomized clinical trial evaluating mETDRS versus normal or high-density micropulse photocoagulation for diabetic macular edema","volume":"52","author":"Lavinsky","year":"2011","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1128\/MCB.01275-12","article-title":"Hsp70 Is a Novel Posttranscriptional Regulator of Gene Expression That Binds and Stabilizes Selected mRNAs Containing AU-Rich Elements","volume":"33","author":"Kishor","year":"2013","journal-title":"Mol. Cell. Biol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/S0006-2952(99)00299-3","article-title":"Heat shock factors and the control of the stress response","volume":"59","author":"Santoro","year":"2000","journal-title":"Biochem. Pharmacol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.exer.2014.04.014","article-title":"Protective effect of a laser-induced sub-lethal temperature rise on RPE cells from oxidative stress","volume":"124","author":"Iwami","year":"2014","journal-title":"Exp. Eye Res."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Fan, B., Sun, Y.J., Liu, S.Y., Che, L., and Li, G.Y. (2017). Neuroprotective strategy in retinal degeneration: Suppressing ER stress-induced cell death via inhibition of the mTOR signal. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18010201"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1146\/annurev-pathol-012414-040508","article-title":"The Roles of Cellular and Organismal Aging in the Development of Late-Onset Maladies","volume":"10","author":"Volovik","year":"2015","journal-title":"Annu. Rev. Pathol. Mech. Dis."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2446","DOI":"10.1016\/j.bbamem.2012.04.008","article-title":"Protein-oxidized phospholipid interactions in cellular signaling for cell death: From biophysics to clinical correlations","volume":"1818","author":"Kinnunen","year":"2012","journal-title":"Biochim. Biophys. Acta (BBA) Biomembr."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1167\/iovs.05-0976","article-title":"Proteomics of the retinal pigment epithelium reveals altered protein expression at progressive stages of age-related macular degeneration","volume":"47","author":"Nordgaard","year":"2006","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"2848","DOI":"10.1167\/iovs.07-1352","article-title":"Mitochondrial proteomics of the retinal pigment epithelium at progressive stages of age-related macular degeneration","volume":"49","author":"Nordgaard","year":"2008","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1016\/j.ajo.2006.12.006","article-title":"Changes in Select Redox Proteins of the Retinal Pigment Epithelium in Age-related Macular Degeneration","volume":"143","author":"Decanini","year":"2007","journal-title":"Am. J. Ophthalmol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"4888","DOI":"10.1167\/iovs.05-1328","article-title":"Thermal injury induces heat shock protein in the optic nerve head in vivo","volume":"47","author":"Kim","year":"2006","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"6591","DOI":"10.1167\/iovs.15-16476","article-title":"Hypoxia-inducible factor-1\u03b1 Is associated with sprouting angiogenesis in the murine laser-induced Choroidal neovascularization model","volume":"56","author":"Tunik","year":"2015","journal-title":"Investig. Ophthalmol. Vis. Sci."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/6\/2063\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:07:35Z","timestamp":1760173655000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/6\/2063"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,3,17]]},"references-count":60,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2020,3]]}},"alternative-id":["ijms21062063"],"URL":"https:\/\/doi.org\/10.3390\/ijms21062063","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2020,3,17]]}}}