{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T02:17:27Z","timestamp":1775096247548,"version":"3.50.1"},"reference-count":64,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2021,12,30]],"date-time":"2021-12-30T00:00:00Z","timestamp":1640822400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["PTDC\/QUI-QUI\/122900\/2010"],"award-info":[{"award-number":["PTDC\/QUI-QUI\/122900\/2010"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["UID\/NEU\/04539\/2013"],"award-info":[{"award-number":["UID\/NEU\/04539\/2013"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["UID\/QUI\/00313\/2019"],"award-info":[{"award-number":["UID\/QUI\/00313\/2019"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["SFRH\/BD\/137991\/2018"],"award-info":[{"award-number":["SFRH\/BD\/137991\/2018"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["RECI\/QEQ- 551 QFI\/0168\/2012"],"award-info":[{"award-number":["RECI\/QEQ- 551 QFI\/0168\/2012"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["UID\/QUI\/00313\/2019"],"award-info":[{"award-number":["UID\/QUI\/00313\/2019"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011929","name":"Programa Operacional Tem\u00e1tico Factores de Competitividade","doi-asserted-by":"publisher","award":["CENTRO-07-CT62-FEDER-002012"],"award-info":[{"award-number":["CENTRO-07-CT62-FEDER-002012"]}],"id":[{"id":"10.13039\/501100011929","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Several degenerative amyloid diseases, with no fully effective treatment, affect millions of people worldwide. These pathologies\u2014amyloidoses\u2014are known to be associated with the formation of ordered protein aggregates and highly stable and insoluble amyloid fibrils, which are deposited in multiple tissues and organs. The disruption of preformed amyloid aggregates and fibrils is one possible therapeutic strategy against amyloidosis; however, only a few compounds have been identified as possible fibril disruptors in vivo to date. To properly identify chemical compounds as potential fibril disruptors, a reliable, fast, and economic screening protocol must be developed. For this purpose, three amyloid fibril formation protocols using transthyretin (TTR), a plasma protein involved in several amyloidoses, were studied using thioflavin-T fluorescence assays, circular dichroism (CD), turbidity, dynamic light scattering (DLS), and transmission electron microscopy (TEM), in order to characterize and select the most appropriate fibril formation protocol. Saturation transfer difference nuclear magnetic resonance spectroscopy (STD NMR) was successfully used to study the interaction of doxycycline, a known amyloid fibril disruptor, with preformed wild-type TTR (TTRwt) aggregates and fibrils. DLS and TEM were also used to characterize the effect of doxycycline on TTRwt amyloid species disaggregation. A comparison of the TTR amyloid morphology formed in different experimental conditions is also presented.<\/jats:p>","DOI":"10.3390\/ijms23010391","type":"journal-article","created":{"date-parts":[[2021,12,30]],"date-time":"2021-12-30T21:41:21Z","timestamp":1640900481000},"page":"391","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Searching for the Best Transthyretin Aggregation Protocol to Study Amyloid Fibril Disruption"],"prefix":"10.3390","volume":"23","author":[{"given":"Elisabete","family":"Ferreira","sequence":"first","affiliation":[{"name":"Chemistry Department and Coimbra Chemistry Centre-Institute of Molecular Sciences (CQC-IMS), University of Coimbra, 3004-535 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4097-2766","authenticated-orcid":false,"given":"Zaida L.","family":"Almeida","sequence":"additional","affiliation":[{"name":"Chemistry Department and Coimbra Chemistry Centre-Institute of Molecular Sciences (CQC-IMS), University of Coimbra, 3004-535 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3578-0134","authenticated-orcid":false,"given":"Pedro F.","family":"Cruz","sequence":"additional","affiliation":[{"name":"Chemistry Department and Coimbra Chemistry Centre-Institute of Molecular Sciences (CQC-IMS), University of Coimbra, 3004-535 Coimbra, Portugal"}]},{"given":"Marta","family":"Silva e Sousa","sequence":"additional","affiliation":[{"name":"Chemistry Department and Coimbra Chemistry Centre-Institute of Molecular Sciences (CQC-IMS), University of Coimbra, 3004-535 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0532-4242","authenticated-orcid":false,"given":"Paula","family":"Ver\u00edssimo","sequence":"additional","affiliation":[{"name":"Centre for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9128-2557","authenticated-orcid":false,"given":"Rui M. M.","family":"Brito","sequence":"additional","affiliation":[{"name":"Chemistry Department and Coimbra Chemistry Centre-Institute of Molecular Sciences (CQC-IMS), University of Coimbra, 3004-535 Coimbra, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Almeida, Z.L., and Brito, R.M.M. (2020). Structure and aggregation mechanisms in amyloids. Molecules, 25.","DOI":"10.3390\/molecules25051195"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"678","DOI":"10.1007\/s00109-003-0464-5","article-title":"Protein aggregation and aggregate toxicity: New insights into protein folding, misfolding diseases and biological evolution","volume":"81","author":"Stefani","year":"2003","journal-title":"J. Mol. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1038\/nrm3810","article-title":"The amyloid state and its association with protein misfolding diseases","volume":"15","author":"Knowles","year":"2014","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"755","DOI":"10.1038\/s41580-018-0060-8","article-title":"A new era for understanding amyloid structures and disease","volume":"19","author":"Iadanza","year":"2018","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1146\/annurev.biochem.75.101304.123901","article-title":"Protein misfolding, functional amyloid, and human disease","volume":"75","author":"Chiti","year":"2006","journal-title":"Annu. Rev. Biochem."},{"key":"ref_6","first-page":"487","article-title":"Fluorescent stains, with special reference to amyloid and connective tissues","volume":"68","author":"Vassar","year":"1959","journal-title":"Arch. Pathol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1080\/07853890701842988","article-title":"Senile systemic amyloidosis affects 25% of the very aged and associates with genetic variation in alpha2-macroglobulin and tau: A population-based autopsy study","volume":"40","author":"Tanskanen","year":"2008","journal-title":"Ann. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1533","DOI":"10.1038\/modpathol.2011.117","article-title":"Clinicopathological features of senile systemic amyloidosis: An ante- and post-mortem study","volume":"24","author":"Ueda","year":"2011","journal-title":"Mod. Pathol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"4499","DOI":"10.1073\/pnas.75.9.4499","article-title":"Amyloid fibril protein related to prealbumin in familial amyloidotic polyneuropathy","volume":"75","author":"Costa","year":"1978","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1056\/NEJM199702133360703","article-title":"Variant-sequence transthyretin (isoleucine 122) in late-onset cardiac amyloidosis in black Americans","volume":"336","author":"Jacobson","year":"1997","journal-title":"N. Engl. J. Med."},{"key":"ref_11","first-page":"351","article-title":"Leptomeningeal amyloid and variant transthyretins","volume":"148","author":"Benson","year":"1996","journal-title":"Am. J. Pathol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1051","DOI":"10.1212\/01.wnl.0000178983.20975.af","article-title":"Neuroradiologic and clinicopathologic features of oculoleptomeningeal type amyloidosis","volume":"65","author":"Nakamura","year":"2005","journal-title":"Neurology"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1002\/mus.20821","article-title":"The molecular biology and clinical features of amyloid neuropathy","volume":"36","author":"Benson","year":"2007","journal-title":"Muscle Nerve"},{"key":"ref_14","unstructured":"(2021, April 16). Mutations in Hereditary Amyloidosis. Available online: http:\/\/amyloidosismutations.com\/mut-attr.php."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/0014-5793(91)80049-9","article-title":"Studies on the synthesis and secretion of transthyretin by the human hepatoma cell line Hep G2","volume":"287","author":"Blaner","year":"1991","journal-title":"FEBS Lett."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/0304-3940(86)90037-6","article-title":"High levels of messenger RNA for transthyretin (prealbumin) in human choroid plexus","volume":"66","author":"Dickson","year":"1986","journal-title":"Neurosci. Lett."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1835","DOI":"10.1021\/bi00173a029","article-title":"Synthesis and secretion of retinol-binding protein and transthyretin by cultured retinal pigment epithelium","volume":"33","author":"Ong","year":"1994","journal-title":"Biochemistry"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Gi\u00e3o, T., Saavedra, J., Cotrina, E., Quintana, J., Llop, J., Arsequell, G., and Cardoso, I. (2020). Undiscovered roles for transthyretin: From a transporter protein to a new therapeutic target for Alzheimer\u2019s disease. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21062075"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1016\/j.bbapap.2008.10.016","article-title":"Protein aggregation kinetics, mechanism, and curve-fitting: A review of the literature","volume":"1794","author":"Morris","year":"2009","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"7255","DOI":"10.1039\/C4CP04549A","article-title":"A look into amyloid formation by transthyretin: Aggregation pathway and a novel kinetic model","volume":"17","author":"Faria","year":"2015","journal-title":"Phys. Chem. Chem. Phys."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"6470","DOI":"10.1021\/bi952501g","article-title":"The acid-mediated denaturation pathway of transthyretin yields a conformational intermediate that can self-assemble into amyloid","volume":"35","author":"Lai","year":"1996","journal-title":"Biochemistry"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"32943","DOI":"10.1074\/jbc.274.46.32943","article-title":"The tetrameric protein transthyretin dissociates to a non-native monomer in solution. A novel model for amyloidogenesis","volume":"274","author":"Quintas","year":"1999","journal-title":"J. Biol. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"27207","DOI":"10.1074\/jbc.M101024200","article-title":"Tetramer dissociation and monomer partial unfolding precedes protofibril formation in amyloidogenic transthyretin variants","volume":"276","author":"Quintas","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Frangolho, A., Correia, B.E., Vaz, D.C., Almeida, Z.L., and Brito, R.M.M. (2020). Oligomerization Profile of Human Transthyretin Variants with Distinct Amyloidogenicity. Molecules, 25.","DOI":"10.3390\/molecules25235698"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2304","DOI":"10.2174\/092986712800269236","article-title":"Transthyretin Deposition in Familial Amyloidotic Polyneuropathy","volume":"19","author":"Saraiva","year":"2012","journal-title":"Curr. Med. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"11070","DOI":"10.1021\/bi201365r","article-title":"Dissecting the structure, thermodynamic stability, and aggregation properties of the A25T transthyretin (A25T-TTR) variant involved in leptomeningeal amyloidosis: Identifying protein partners that co-aggregate during A25T-TTR fibrillogenesis in cerebrospinal fluid","volume":"50","author":"Azevedo","year":"2011","journal-title":"Biochemistry"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"9629","DOI":"10.1073\/pnas.1121005109","article-title":"Tafamidis, a potent and selective transthyretin kinetic stabilizer that inhibits the amyloid cascade","volume":"109","author":"Bulawa","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1006\/jsbi.2000.4273","article-title":"Review: TTR amyloidosis-structural features leading to protein aggregation and their implications on therapeutic strategies","volume":"130","author":"Damas","year":"2000","journal-title":"J. Struct. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1186\/2047-9158-3-19","article-title":"Recent advances in transthyretin amyloidosis therapy","volume":"3","author":"Ueda","year":"2014","journal-title":"Transl. Neurodegener."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1002\/ejhf.1695","article-title":"Emerging therapies in transthyretin amyloidosis\u2014A new wave of hope after years of stagnancy?","volume":"22","author":"Butler","year":"2020","journal-title":"Eur. J. Heart Fail."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1080\/13506129.2016.1191458","article-title":"Suppressing transthyretin production in mice, monkeys and humans using 2nd-Generation antisense oligonucleotides","volume":"23","author":"Ackermann","year":"2016","journal-title":"Amyloid"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1056\/NEJMoa1716793","article-title":"Inotersen treatment for patients with Hereditary transthyretin amyloidosis","volume":"379","author":"Benson","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1374","DOI":"10.1111\/ene.14285","article-title":"Early data on long-term efficacy and safety of inotersen in patients with hereditary transthyretin amyloidosis: A 2-year update from the open-label extension of the NEURO-TTR trial","volume":"27","author":"Brannagan","year":"2020","journal-title":"Eur. J. Neurol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13023-015-0326-6","article-title":"Efficacy and safety of patisiran for familial amyloidotic polyneuropathy: A phase II multi-dose study","volume":"10","author":"Suhr","year":"2015","journal-title":"Orphanet. J. Rare Dis."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Adams, D., Suhr, O.B., Dyck, P.J., Litchy, W.J., Leahy, R.G., Chen, J., Gollob, J., and Coelho, T. (2017). Trial design and rationale for APOLLO, a Phase 3, placebo-controlled study of patisiran in patients with hereditary ATTR amyloidosis with polyneuropathy. BMC Neurol., 17.","DOI":"10.1186\/s12883-017-0948-5"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1002\/jcph.1553","article-title":"Pharmacokinetics of Patisiran, the First Approved RNA Interference Therapy in Patients with Hereditary Transthyretin-Mediated Amyloidosis","volume":"60","author":"Zhang","year":"2019","journal-title":"J. Clin. Pharmacol."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Ferreira, N., Saraiva, M.J., and Almeida, M.R. (2012). Epigallocatechin-3-gallate as a potential therapeutic drug for TTR-related amyloidosis: \u201cin vivo\u201d evidence from FAP mice models. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0029933"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1007\/s00392-012-0463-z","article-title":"Green tea halts progression of cardiac transthyretin amyloidosis: An observational report","volume":"101","author":"Kristen","year":"2012","journal-title":"Clin. Res. Cardiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2959","DOI":"10.1073\/pnas.92.7.2959","article-title":"Interaction of the anthracycline 4\u2032-iodo-4\u2032-deoxydoxorubicin with amyloid fibrils: Inhibition of amyloidogenesis","volume":"92","author":"Merlini","year":"1995","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1096\/fj.02-0764com","article-title":"4\u2032-iodo-4\u2032-Deoxydoxorubicin and tetracyclines disrupt transthyretin amyloid fibrils in vitro producing noncytotoxic species: Screening for TTR fibril disrupters","volume":"17","author":"Cardoso","year":"2003","journal-title":"FASEB J."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1155\/2008\/549872","article-title":"Extracellular matrix markers for disease progression and follow-up of therapies in familial amyloid polyneuropathy V30M TTR-related","volume":"25","author":"Cardoso","year":"2008","journal-title":"Dis. Markers"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1096\/fj.05-4509com","article-title":"Doxycycline disrupts transthyretin amyloid: Evidence from studies in a FAP transgenic mice model","volume":"20","author":"Cardoso","year":"2006","journal-title":"FASEB J."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"34","DOI":"10.3109\/13506129.2012.678508","article-title":"Doxycycline plus tauroursodeoxycholic acid for transthyretin amyloidosis: A phase II study","volume":"19","author":"Obici","year":"2012","journal-title":"Amyloid"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2424","DOI":"10.1016\/j.febslet.2011.06.030","article-title":"Natural polyphenols inhibit different steps of the process of transthyretin (TTR) amyloid fibril formation","volume":"585","author":"Ferreira","year":"2011","journal-title":"FEBS Lett."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"11391","DOI":"10.1021\/jf902664f","article-title":"Tea catechins induce the conversion of preformed lysozyme amyloid fibrils to amorphous aggregates","volume":"57","author":"He","year":"2009","journal-title":"J. Agric. Food Chem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"404","DOI":"10.1016\/S0014-5793(00)02380-2","article-title":"Anti-amyloidogenic activity of tetracyclines: Studies in vitro","volume":"487","author":"Forloni","year":"2001","journal-title":"FEBS Lett."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"2795","DOI":"10.1016\/S0006-3495(03)70009-5","article-title":"The pH-dependent stability of wild-type and mutant transthyretin oligomers","volume":"84","author":"Skoulakis","year":"2003","journal-title":"Biophys. J."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/S0301-4622(00)00199-X","article-title":"Comparative calorimetric study of non-amyloidogenic and amyloidogenic variants of the homotetrameric protein transthyretin","volume":"88","author":"Shnyrov","year":"2000","journal-title":"Biophys. Chem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"11442","DOI":"10.1021\/bi011194d","article-title":"An engineered transthyretin monomer that is nonamyloidogenic, unless it is partially denatured","volume":"40","author":"Jiang","year":"2001","journal-title":"Biochemistry"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"8654","DOI":"10.1021\/bi00151a036","article-title":"Partial denaturation of transthyretin is sufficient for amyloid fibril formation in vitro","volume":"31","author":"Colon","year":"1992","journal-title":"Biochemistry"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1999","DOI":"10.1111\/j.1742-4658.2009.06936.x","article-title":"Stability and fibril formation properties of human and fish transthyretin, and of the Escherichia coli transthyretin-related protein","volume":"276","author":"Lundberg","year":"2009","journal-title":"FEBS J."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/j.cardfail.2019.01.006","article-title":"Clinical experience with the use of doxycycline and ursodeoxycholic acid for the treatment of transthyretin cardiac amyloidosis","volume":"25","author":"Karlstedt","year":"2019","journal-title":"J. Card. Fail."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1002\/pro.3105","article-title":"(S)Pinning down protein interactions by NMR","volume":"26","author":"Teilum","year":"2017","journal-title":"Protein Sci."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"4200","DOI":"10.1529\/biophysj.104.049700","article-title":"Detection and characterization of aggregates, prefibrillar amyloidogenic oligomers, and protofibrils using fluorescence spectroscopy","volume":"88","author":"Lindgren","year":"2005","journal-title":"Biophys. J."},{"key":"ref_55","first-page":"349","article-title":"Amyloid Formation by Transthyretin: From Protein Stability to Protein Aggregation","volume":"3","author":"Brito","year":"2003","journal-title":"Curr. Med. Chem. Endocr. Metab. Agents"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1021\/cc049849s","article-title":"Kinetic assay for high-throughput screening of in vitro transthyretin amyloid fibrillogenesis inhibitors","volume":"7","author":"Dolado","year":"2005","journal-title":"J. Comb. Chem."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"2343","DOI":"10.2174\/092986712800269281","article-title":"Planas, Methods to evaluate the inhibition of TTR fibrillogenesis induced by small ligands","volume":"19","author":"Arsequell","year":"2012","journal-title":"Curr. Med. Chem."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"823","DOI":"10.1016\/j.ejmech.2016.02.074","article-title":"A novel bis-furan scaffold for transthyretin stabilization and amyloid inhibition","volume":"121","author":"Almeida","year":"2016","journal-title":"Eur. J. Med. Chem."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"990","DOI":"10.1021\/ed101169t","article-title":"Saturation-transfer difference (STD) NMR: A simple and fast method for ligand screening and characterization of protein binding","volume":"88","author":"Viegas","year":"2011","journal-title":"J. Chem. Educ."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1080\/13506129.2016.1277695","article-title":"Doxycycline-tauroursodeoxycholic acid treatment: Effects in the heart of a transthyretin V30M transgenic mouse model","volume":"24","author":"Teixeira","year":"2017","journal-title":"Amyloid"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"2307","DOI":"10.1046\/j.1432-1327.2000.01237.x","article-title":"Aprotinin binding to amyloid fibrils","volume":"267","author":"Cardoso","year":"2000","journal-title":"Eur. J. Biochem."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"W315","DOI":"10.1093\/nar\/gky497","article-title":"BeStSel: A web server for accurate protein secondary structure prediction and fold recognition from the circular dichroism spectra","volume":"46","author":"Micsonai","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_63","first-page":"36","article-title":"Image processing with ImageJ","volume":"11","author":"Ram","year":"2004","journal-title":"Biophotonics Int."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"1935","DOI":"10.1016\/S0002-9440(10)64469-0","article-title":"Evidence for early cytotoxic aggregates in transgenic mice for human transthyretin Leu55Pro","volume":"161","author":"Sousa","year":"2002","journal-title":"Am. J. Pathol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/1\/391\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:55:56Z","timestamp":1760169356000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/1\/391"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,30]]},"references-count":64,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2022,1]]}},"alternative-id":["ijms23010391"],"URL":"https:\/\/doi.org\/10.3390\/ijms23010391","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,30]]}}}