{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T02:31:42Z","timestamp":1768444302432,"version":"3.49.0"},"reference-count":49,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2020,8,19]],"date-time":"2020-08-19T00:00:00Z","timestamp":1597795200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Coordinaci\u00f3n de Posgrado e Investigaci\u00f3n-UABC","award":["106\/2\/N\/57\/1 (1983)"],"award-info":[{"award-number":["106\/2\/N\/57\/1 (1983)"]}]},{"name":"Fondo Sectorial de Investigaci\u00f3n para la Educaci\u00f3n CB 2017-2018","award":["A1-S-28653\/SEP\/CONACYT"],"award-info":[{"award-number":["A1-S-28653\/SEP\/CONACYT"]}]},{"name":"21a. Convocatoria Interna de Apoyo a Proyectos de Investigaci\u00f3n","award":["Coordinaci\u00f3n General de Investigaci\u00f3n y Posgrado\/UABC"],"award-info":[{"award-number":["Coordinaci\u00f3n General de Investigaci\u00f3n y Posgrado\/UABC"]}]},{"name":"DGTIC-UNAM (supercomputing cluster MIZTLI)","award":["LANCAD-UNAM-DGTIC-352"],"award-info":[{"award-number":["LANCAD-UNAM-DGTIC-352"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Human islet amyloid polypeptide (hIAPP) corresponds to a 37-residue hormone present in insulin granules that maintains a high propensity to form \u03b2-sheet structures during co-secretion with insulin. Previously, employing a biomimetic approach, we proposed a panel of optimized IAPP sequences with only one residue substitution that shows the capability to reduce amyloidogenesis. Taking into account that specific membrane lipids have been considered as a key factor in the induction of cytotoxicity, in this study, following the same design strategy, we characterize the effect of a series of lipids upon several polypeptide domains that show the highest aggregation propensity. The characterization of the C-native segment of hIAPP (residues F23-Y37), together with novel variants F23R and I26A allowed us to demonstrate an effect upon the formation of \u03b2-sheet structures. Our results suggest that zwitterionic phospholipids promote adsorption of the C-native segments at the lipid-interface and \u03b2-sheet formation with the exception of the F23R variant. Moreover, the presence of cholesterol did not modify this behavior, and the \u03b2-sheet structural transitions were not registered when the N-terminal domain of hIAPP (K1-S20) was characterized. Considering that insulin granules are enriched in phosphatidylserine (PS), the property of lipid vesicles containing negatively charged lipids was also evaluated. We found that these types of lipids promote \u03b2-sheet conformational transitions in both the C-native segment and the new variants. Furthermore, these PS\/peptides arrangements are internalized in Langerhans islet \u03b2-cells, localized in the endoplasmic reticulum, and trigger critical pathways such as unfolded protein response (UPR), affecting insulin secretion. Since this phenomenon was associated with the presence of cytotoxicity on Langerhans islet \u03b2-cells, it can be concluded that the anionic lipid environment and degree of solvation are critical conditions for the stability of segments with the propensity to form \u03b2-sheet structures, a situation that will eventually affect the structural characteristics and stability of IAPP within insulin granules, thus modifying the insulin secretion.<\/jats:p>","DOI":"10.3390\/biom10091201","type":"journal-article","created":{"date-parts":[[2020,8,19]],"date-time":"2020-08-19T09:22:31Z","timestamp":1597828951000},"page":"1201","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Lipid Modulation in the Formation of \u03b2-Sheet Structures. Implications for De Novo Design of Human Islet Amyloid Polypeptide and the Impact on \u03b2-Cell Homeostasis"],"prefix":"10.3390","volume":"10","author":[{"given":"Israel","family":"Mart\u00ednez-Navarro","sequence":"first","affiliation":[{"name":"Departamento de Bioqu\u00edmica, Facultad de Medicina Mexicali, Universidad Aut\u00f3noma de Baja California, Mexicali 21000, Baja California, Mexico"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9400-5683","authenticated-orcid":false,"given":"Ra\u00fal","family":"D\u00edaz-Molina","sequence":"additional","affiliation":[{"name":"Departamento de Bioqu\u00edmica, Facultad de Medicina Mexicali, Universidad Aut\u00f3noma de Baja California, Mexicali 21000, Baja California, Mexico"}]},{"given":"Angel","family":"Pulido-Capiz","sequence":"additional","affiliation":[{"name":"Departamento de Bioqu\u00edmica, Facultad de Medicina Mexicali, Universidad Aut\u00f3noma de Baja California, Mexicali 21000, Baja California, Mexico"},{"name":"Laboratorio de Biolog\u00eda Molecular, Facultad de Medicina Mexicali, Universidad Aut\u00f3noma de Baja California, Mexicali 21000, Baja California, Mexico."}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8007-6993","authenticated-orcid":false,"given":"Jaime","family":"Mas-Oliva","sequence":"additional","affiliation":[{"name":"Instituto de Fisiolog\u00eda Celular, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Ciudad de Mexico 04510, Mexico"}]},{"given":"Ismael","family":"Luna-Reyes","sequence":"additional","affiliation":[{"name":"Instituto de Fisiolog\u00eda Celular, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Ciudad de Mexico 04510, Mexico"}]},{"given":"Eustolia","family":"Rodr\u00edguez-Vel\u00e1zquez","sequence":"additional","affiliation":[{"name":"Facultad de Odontolog\u00eda, Universidad Aut\u00f3noma de Baja California, Tijuana 22390, Mexico"},{"name":"Tecnol\u00f3gico Nacional de M\u00e9xico\/I.T. Tijuana, Centro de Graduados e Investigaci\u00f3n en Qu\u00edmica-Grupo de Biomateriales y Nanomedicina, Tijuana 22510, Mexico"}]},{"given":"Ignacio A.","family":"Rivero","sequence":"additional","affiliation":[{"name":"Tecnol\u00f3gico Nacional de M\u00e9xico\/Instituto Tecnol\u00f3gico de Tijuana, Centro de Graduados e Investigaci\u00f3n en Qu\u00edmica, Tijuana 22510, Baja California, Mexico"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8950-2079","authenticated-orcid":false,"given":"Marco A.","family":"Ramos-Ibarra","sequence":"additional","affiliation":[{"name":"Facultad de Ciencias Qu\u00edmicas e Ingenier\u00eda, Universidad Aut\u00f3noma de Baja California, Tijuana 22390, Baja California, Mexico"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3301-9619","authenticated-orcid":false,"given":"Manuel","family":"Alatorre-Meda","sequence":"additional","affiliation":[{"name":"C\u00e1tedras CONACyT- Tecnol\u00f3gico Nacional de M\u00e9xico\/I.T. Tijuana, Centro de Graduados e Investigaci\u00f3n en Qu\u00edmica-Grupo de Biomateriales y Nanomedicina, Tijuana 22510, Mexico"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9421-3730","authenticated-orcid":false,"given":"Victor","family":"Garc\u00eda-Gonz\u00e1lez","sequence":"additional","affiliation":[{"name":"Departamento de Bioqu\u00edmica, Facultad de Medicina Mexicali, Universidad Aut\u00f3noma de Baja California, Mexicali 21000, Baja California, Mexico"}]}],"member":"1968","published-online":{"date-parts":[[2020,8,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1179","DOI":"10.1016\/0140-6736(92)90785-2","article-title":"Amylin concentrations and glucose control","volume":"339","author":"Koda","year":"1992","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1093\/clinchem\/42.4.576","article-title":"Development of sensitive immunoassays to detect amylin and amylin-like peptides in unextracted plasma","volume":"42","author":"Percy","year":"1996","journal-title":"Clin. Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1514","DOI":"10.2337\/diab.42.10.1514","article-title":"Regulation of islet amyloid polypeptide in human pancreatic islets","volume":"42","author":"Novials","year":"1993","journal-title":"Diabetes"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"7308","DOI":"10.1021\/jp8106827","article-title":"Conformation preorganization: Effects of S20G mutation on the structure of human islet amyloid polypeptide segment","volume":"113","author":"Xu","year":"2009","journal-title":"J. Phys. Chem. B"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"574","DOI":"10.1007\/s00125-003-1068-x","article-title":"Amylin gene promoter mutations predispose to Type 2 diabetes in New Zealand Maori","volume":"46","author":"Poa","year":"2003","journal-title":"Diabetologia"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Berhanu, W.M., and Hansmann, U.H. (2014). Inter-species cross-seeding: Stability and assembly of rat-human amylin aggregates. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0097051"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"331","DOI":"10.3389\/fendo.2018.00331","article-title":"Modulation of Amyloidogenesis Controlled by the C-Terminal Domain of Islet Amyloid Polypeptide Shows New Functions on Hepatocyte Cholesterol Metabolism","volume":"9","author":"Rivero","year":"2018","journal-title":"Front. Endocrinol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"16091","DOI":"10.1038\/srep16091","article-title":"The C-terminal Domain Supports a Novel Function for CETPI as a New Plasma Lipopolysaccharide-Binding Protein","volume":"5","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.jsb.2014.02.002","article-title":"Key structural arrangements at the C-terminus domain of CETP suggest a potential mechanism for lipid-transfer activity","volume":"186","author":"Brocos","year":"2014","journal-title":"J. Struct. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.bbrc.2013.03.067","article-title":"Amyloid fibril formation of peptides derived from the C-terminus of CETP modulated by lipids","volume":"434","year":"2013","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"17193","DOI":"10.3390\/ijms160817193","article-title":"Protein Folding and Mechanisms of Proteostasis","volume":"16","year":"2015","journal-title":"Int. J. Mol. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2019","DOI":"10.3390\/ijms12032019","article-title":"Amyloidogenic properties of a D\/N mutated 12 amino acid fragment of the C-terminal domain of the Cholesteryl-Ester Transfer Protein (CETP)","volume":"12","year":"2011","journal-title":"Int. J. Mol. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1007\/s11010-009-0105-6","article-title":"Disorder-to-order conformational transitions in protein structure and its relationship to disease","volume":"330","author":"Moreno","year":"2009","journal-title":"Mol. Cell. Biochem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"9496","DOI":"10.1021\/bi060579z","article-title":"Conserved and cooperative assembly of membrane-bound alpha-helical states of islet amyloid polypeptide","volume":"45","author":"Knight","year":"2006","journal-title":"Biochemistry"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"12113","DOI":"10.1021\/bi050840w","article-title":"Lipid membranes modulate the structure of islet amyloid polypeptide","volume":"44","author":"Jayasinghe","year":"2005","journal-title":"Biochemistry"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1016\/j.tem.2010.06.003","article-title":"The insulin secretory granule as a signaling hub","volume":"21","author":"Suckale","year":"2010","journal-title":"Trends Endocrinol. Metab."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"11075","DOI":"10.1074\/jbc.M114.628420","article-title":"Characterization of phospholipids in insulin secretory granules and mitochondria in pancreatic beta cells and their changes with glucose stimulation","volume":"290","author":"MacDonald","year":"2015","journal-title":"J. Biol. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Saito Michiko, S.Y. (2019). ER Stress, Secretory Granule Biogenesis, and Insulin. Ultimate Guide to Insulin, IntechOpen.","DOI":"10.5772\/intechopen.76131"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1083\/jcb.200907074","article-title":"Membrane expansion alleviates endoplasmic reticulum stress independently of the unfolded protein response","volume":"187","author":"Schuck","year":"2009","journal-title":"J. Cell. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"R1","DOI":"10.1530\/JME-15-0306","article-title":"Endoplasmic reticulum stress and the unfolded protein response in pancreatic islet inflammation","volume":"57","author":"Meyerovich","year":"2016","journal-title":"J. Mol. Endocrinol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"4158","DOI":"10.1002\/jcb.27701","article-title":"Protein translation associated to PERK arm is a new target for regulation of metainflammation: A connection with hepatocyte cholesterol","volume":"120","year":"2019","journal-title":"J. Cell. Biochem."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Acosta-Montano, P., Rodriguez-Velazquez, E., Ibarra-Lopez, E., Frayde-Gomez, H., Mas-Oliva, J., Delgado-Coello, B., Rivero, I.A., Alatorre-Meda, M., Aguilera, J., and Guevara-Olaya, L. (2019). Fatty Acid and Lipopolysaccharide Effect on Beta Cells Proteostasis and its Impact on Insulin Secretion. Cells, 8.","DOI":"10.3390\/cells8080884"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Conchillo-Sole, O., de Groot, N.S., Aviles, F.X., Vendrell, J., Daura, X., and Ventura, S. (2007). AGGRESCAN: A server for the prediction and evaluation of \"hot spots\" of aggregation in polypeptides. BMC Bioinform., 8.","DOI":"10.1186\/1471-2105-8-65"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1146\/annurev.bb.09.060180.002343","article-title":"Comparative properties and methods of preparation of lipid vesicles (liposomes)","volume":"9","author":"Szoka","year":"1980","journal-title":"Annu. Rev. Biophys. Bioeng."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"6127","DOI":"10.1021\/acs.jctc.8b00391","article-title":"Avoiding False Positive Conclusions in Molecular Simulation: The Importance of Replicas","volume":"14","author":"Knapp","year":"2018","journal-title":"J. Chem. Theory Comput."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6994","DOI":"10.1021\/bi2007564","article-title":"Structural role of compensatory amino acid replacements in the alpha-synuclein protein","volume":"50","author":"Losasso","year":"2011","journal-title":"Biochemistry"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1007\/978-1-0716-0138-9_11","article-title":"Methodology for Subcellular Fractionation and MicroRNA Examination of Mitochondria, Mitochondria Associated ER Membrane (MAM), ER, and Cytosol from Human Brain","volume":"2063","author":"Prajapati","year":"2020","journal-title":"Methods Mol. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"946037","DOI":"10.1155\/2015\/946037","article-title":"Charge-Based Inhibitors of Amylin Fibrillization and Toxicity","volume":"2015","author":"Patil","year":"2015","journal-title":"J. Diabetes Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"13748","DOI":"10.1021\/bi0011330","article-title":"Amyloid fibril formation by A beta 16-22, a seven-residue fragment of the Alzheimer\u2019s beta-amyloid peptide, and structural characterization by solid state NMR","volume":"39","author":"Balbach","year":"2000","journal-title":"Biochemistry"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1014","DOI":"10.1002\/prot.24795","article-title":"Complete characterization of the mutation landscape reveals the effect on amylin stability and amyloidogenicity","volume":"83","author":"Smaoui","year":"2015","journal-title":"Proteins"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1016\/S0091-679X(08)00813-3","article-title":"Nano-scale imaging and dynamics of amylin-membrane interactions and its implication in type II diabetes mellitus","volume":"90","author":"Cho","year":"2008","journal-title":"Methods Cell. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"15598","DOI":"10.1021\/ja205007j","article-title":"Low pH acts as inhibitor of membrane damage induced by human islet amyloid polypeptide","volume":"133","author":"Khemtemourian","year":"2011","journal-title":"J. Am. Chem. Soc."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1016\/j.jmb.2009.08.055","article-title":"Cholesterol regulates assembly of human islet amyloid polypeptide on model membranes","volume":"393","author":"Cho","year":"2009","journal-title":"J Mol Biol"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1038\/35036052","article-title":"Lipid rafts and signal transduction","volume":"1","author":"Simons","year":"2000","journal-title":"Nat. Rev. Mol. Cell. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2031","DOI":"10.1021\/acs.biochem.5b00507","article-title":"Conformational Dynamics of the Human Islet Amyloid Polypeptide in a Membrane Environment: Toward the Aggregation Prone Form","volume":"55","author":"Skeby","year":"2016","journal-title":"Biochemistry"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1039\/C1CS15112F","article-title":"Misfolded proteins in Alzheimer\u2019s disease and type II diabetes","volume":"41","author":"DeToma","year":"2012","journal-title":"Chem. Soc. Rev."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1038\/82815","article-title":"Mechanisms of amyloidogenesis","volume":"7","author":"Kelly","year":"2000","journal-title":"Nat. Struct. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"051922","DOI":"10.1103\/PhysRevE.84.051922","article-title":"Effects of cholesterol on pore formation in lipid bilayers induced by human islet amyloid polypeptide fragments: A coarse-grained molecular dynamics study","volume":"84","author":"Xu","year":"2011","journal-title":"Phys. Rev. E"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1673","DOI":"10.1016\/j.bbamem.2010.11.009","article-title":"On the role of anionic lipids in charged protein interactions with membranes","volume":"1808","author":"Vorobyov","year":"2011","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e1282022","DOI":"10.1080\/15592324.2017.1282022","article-title":"Anionic lipids and the maintenance of membrane electrostatics in eukaryotes","volume":"12","author":"Platre","year":"2017","journal-title":"Plant Signal. Behav."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1021\/jacs.6b06985","article-title":"Membrane Permeation versus Amyloidogenicity: A Multitechnique Study of Islet Amyloid Polypeptide Interaction with Model Membranes","volume":"139","author":"Martel","year":"2017","journal-title":"J. Am. Chem. Soc."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1016\/j.colsurfb.2018.12.066","article-title":"Dissecting the effects of free fatty acids on the thermodynamic stability of complex model membranes mimicking insulin secretory granules","volume":"176","author":"Saitta","year":"2019","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1021\/acs.biochem.6b01016","article-title":"Islet Amyloid Polypeptide Membrane Interactions: Effects of Membrane Composition","volume":"56","author":"Zhang","year":"2017","journal-title":"Biochemistry"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Jang, I., Pottekat, A., Poothong, J., Yong, J., Lagunas-Acosta, J., Charbono, A., Chen, Z., Scheuner, D.L., Liu, M., and Itkin-Ansari, P. (2019). PDIA1\/P4HB is required for efficient proinsulin maturation and ss cell health in response to diet induced obesity. eLife, 8.","DOI":"10.7554\/eLife.44528"},{"key":"ref_45","first-page":"70","article-title":"Protein disulfide isomerase a multifunctional protein with multiple physiological roles","volume":"2","author":"Mutus","year":"2014","journal-title":"Front Chem."},{"key":"ref_46","unstructured":"Avelino Jorge, G.-G.V., Alatorre-Meda, M., Rodr\u00edguez-Vel\u00e1zquez, E., and Rivero, I.A. (2020). Synthesis of BODIPY-amino acids and the potential applications as specific dyes for the cytoplasm of Langerhans \u03b2-cells, (in review)."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"5249","DOI":"10.1021\/acsomega.9b04313","article-title":"Preparation of Polymeric Films of PVDMA-PEI Functionalized with Fatty Acids for Studying the Adherence and Proliferation of Langerhans beta-Cells","volume":"5","author":"Rivero","year":"2020","journal-title":"ACS Omega"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"129422","DOI":"10.1016\/j.bbagen.2019.129422","article-title":"Fibrillation of human islet amyloid polypeptide and its toxicity to pancreatic beta-cells under lipid environment","volume":"1864","author":"Gao","year":"2020","journal-title":"Biochim. Biophys. Acta Gen. Subj."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1038\/nchem.1293","article-title":"Two-dimensional infrared spectroscopy reveals the complex behaviour of an amyloid fibril inhibitor","volume":"4","author":"Middleton","year":"2012","journal-title":"Nat. Chem."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/10\/9\/1201\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:02:50Z","timestamp":1760176970000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/10\/9\/1201"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,8,19]]},"references-count":49,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2020,9]]}},"alternative-id":["biom10091201"],"URL":"https:\/\/doi.org\/10.3390\/biom10091201","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,8,19]]}}}