Urease de helicobacter pylori : produ??o, marca??o fluorescente e estudo da biodistribui??o por imageamento ?ptico in vivo
Ano de defesa: | 2021 |
---|---|
Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | |
Tipo de documento: | Dissertação |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Pontif?cia Universidade Cat?lica do Rio Grande do Sul
|
Programa de Pós-Graduação: |
Programa de P?s-Gradua??o em Engenharia e Tecnologia de Materiais
|
Departamento: |
Escola Polit?cnica
|
País: |
Brasil
|
Palavras-chave em Português: | |
Palavras-chave em Inglês: | |
Área do conhecimento CNPq: | |
Link de acesso: | http://tede2.pucrs.br/tede2/handle/tede/10070 |
Resumo: | Alzheimer?s Disease (AD) has the presence of brain neurofibrillary tangles ? deposits of the hyperphosphorylated tau protein ? among its histopathological hallmarks. AD patients are more often infected with Helicobacter pylori (a gastric pathogen) than healthy individuals, and it has been reported that an H. pylori culture filtrate induced tau hyperphosphorylation in vitro and in vivo. H. pylori infects about half of the world?s population, and its survival in the human stomach is only viable due to the enzyme urease (H. pylori urease, HPU), possibly involved in AD pathogenesis, although little is known about its in vivo biodistribution. The present work aimed to investigate HPU?s absorption and biodistribution kinetics in the nude mouse (Mus musculus Foxn1nu) model using in vivo optical imaging. For this purpose, HPU was produced, characterized, labeled with fluorophores, and given intraperitoneally to the animals. The protein?s oligomerization states were monitored during weeks, and it displayed stability in the dodecameric form. In silico studies indicated the presence of potentially free amino acid residues in HPU?s surface for fluorescent labeling, and the latter did not affect HPU?s enzymatic activity. The developed methodology was efficient in acquiring in vivo images to monitor the biodistribution and absorption and excretion kinetics of the labeled HPU. Rapid absorption of the protein by the circulatory system from the peritoneal cavity and later excretion by the kidneys was observed, followed by the migration to the bladder ? approximately 40 min after the injection. Further investigations involving chronic treatment with HPU are required to seek deeper clarification on the mechanisms through which the enzyme, produced by a gastric pathogen, may be involved in neuropathogenic processes. |
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Papal?o, Ricardo Meurerhttp://lattes.cnpq.br/1933730859000512Carlini, C?lia Regina Ribeiro da Silvahttp://lattes.cnpq.br/5587901760491970http://lattes.cnpq.br/8980765616740339Higuchi, Kiyo Costa2022-02-01T18:30:21Z2021-08-30http://tede2.pucrs.br/tede2/handle/tede/10070Alzheimer?s Disease (AD) has the presence of brain neurofibrillary tangles ? deposits of the hyperphosphorylated tau protein ? among its histopathological hallmarks. AD patients are more often infected with Helicobacter pylori (a gastric pathogen) than healthy individuals, and it has been reported that an H. pylori culture filtrate induced tau hyperphosphorylation in vitro and in vivo. H. pylori infects about half of the world?s population, and its survival in the human stomach is only viable due to the enzyme urease (H. pylori urease, HPU), possibly involved in AD pathogenesis, although little is known about its in vivo biodistribution. The present work aimed to investigate HPU?s absorption and biodistribution kinetics in the nude mouse (Mus musculus Foxn1nu) model using in vivo optical imaging. For this purpose, HPU was produced, characterized, labeled with fluorophores, and given intraperitoneally to the animals. The protein?s oligomerization states were monitored during weeks, and it displayed stability in the dodecameric form. In silico studies indicated the presence of potentially free amino acid residues in HPU?s surface for fluorescent labeling, and the latter did not affect HPU?s enzymatic activity. The developed methodology was efficient in acquiring in vivo images to monitor the biodistribution and absorption and excretion kinetics of the labeled HPU. Rapid absorption of the protein by the circulatory system from the peritoneal cavity and later excretion by the kidneys was observed, followed by the migration to the bladder ? approximately 40 min after the injection. Further investigations involving chronic treatment with HPU are required to seek deeper clarification on the mechanisms through which the enzyme, produced by a gastric pathogen, may be involved in neuropathogenic processes.A doen?a de Alzheimer (DA) tem a presen?a no c?rebro de emaranhados neurofibrilares ? dep?sitos da prote?na tau hiperfosforilada ? dentre seus marcadores histopatol?gicos. Pacientes com DA apresentam maior incid?ncia de infec??o por Helicobacter pylori (um pat?geno g?strico) do que indiv?duos saud?veis, e foi reportado que um filtrado da cultura de H. pylori induziu a hiperfosforila??o da prote?na tau in vitro e in vivo. O H. pylori infecta cerca de metade da popula??o mundial, e sua sobreviv?ncia no est?mago humano s? ? vi?vel gra?as ? enzima urease (H. pylori urease, HPU), possivelmente envolvida na patog?nese da DA, apesar de haver pouco conhecimento da sua biodistribui??o in vivo. O presente trabalho teve como objetivo investigar a cin?tica de absor??o e biodistribui??o da HPU no modelo camundongo ?nude? (Mus musculus Foxn1nu), utilizando imageamento ?ptico in vivo. Para isso, a HPU foi produzida, caracterizada, marcada com fluor?foros e administrada por via intraperitoneal nos animais. Os estados de oligomeriza??o da prote?na foram monitorados durante semanas, e ela se mostrou est?vel na forma dodecam?rica. Estudos in silico indicaram a presen?a de res?duos de amino?cidos potencialmente livres na superf?cie da HPU para a marca??o fluorescente, e essa n?o afetou sua atividade enzim?tica. A metodologia desenvolvida se mostrou eficiente na aquisi??o de imagens in vivo para monitorar a biodistribui??o e cin?tica de absor??o e excre??o da HPU marcada. Observou-se sua r?pida absor??o pelo sistema circulat?rio a partir da cavidade peritoneal e excre??o pelos rins, migrando para a bexiga ? aproximadamente 40 min ap?s a inje??o. Estudos adicionais envolvendo tratamentos cr?nicos com a HPU s?o necess?rios a fim de buscar maior elucida??o acerca dos mecanismos por meio dos quais a enzima, proveniente de um pat?geno g?strico, pode estar envolvida em processos neuropatog?nicos.Submitted by PPG Engenharia e Tecnologia de Materiais (engenharia.pg.materiais@pucrs.br) on 2022-01-31T18:16:11Z No. of bitstreams: 1 Disserta??o_Kiyo Costa Higuchi.pdf: 4067661 bytes, checksum: 352d8f6c66669a1586d4da3b642fd872 (MD5)Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2022-02-01T18:21:43Z (GMT) No. of bitstreams: 1 Disserta??o_Kiyo Costa Higuchi.pdf: 4067661 bytes, checksum: 352d8f6c66669a1586d4da3b642fd872 (MD5)Made available in DSpace on 2022-02-01T18:30:21Z (GMT). No. of bitstreams: 1 Disserta??o_Kiyo Costa Higuchi.pdf: 4067661 bytes, checksum: 352d8f6c66669a1586d4da3b642fd872 (MD5) Previous issue date: 2021-08-30Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPESapplication/pdfhttp://tede2.pucrs.br:80/tede2/retrieve/183209/Disserta%c3%a7%c3%a3o_Kiyo%20Costa%20Higuchi.pdf.jpgporPontif?cia Universidade Cat?lica do Rio Grande do SulPrograma de P?s-Gradua??o em Engenharia e Tecnologia de MateriaisPUCRSBrasilEscola Polit?cnicaUrease de Helicobacter PyloriImageamento ?ptico in VivoBiodistribui??oFluor?foroBiodistributionFluoroprobeHelicobacter Pylori UreaseIn Vivo Optical ImagingENGENHARIASUrease de helicobacter pylori : produ??o, marca??o fluorescente e estudo da biodistribui??o por imageamento ?ptico in vivoinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisTrabalho n?o apresenta restri??o para publica??o495391460509391966550050060045189710564848268253590462550136975366info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da PUC_RSinstname:Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS)instacron:PUC_RSTHUMBNAILDisserta??o_Kiyo Costa Higuchi.pdf.jpgDisserta??o_Kiyo Costa Higuchi.pdf.jpgimage/jpeg5013http://tede2.pucrs.br/tede2/bitstream/tede/10070/4/Disserta%C3%A7%C3%A3o_Kiyo+Costa+Higuchi.pdf.jpg0b9b43bdb192f653121174bb6c26e381MD54TEXTDisserta??o_Kiyo Costa Higuchi.pdf.txtDisserta??o_Kiyo Costa Higuchi.pdf.txttext/plain156802http://tede2.pucrs.br/tede2/bitstream/tede/10070/3/Disserta%C3%A7%C3%A3o_Kiyo+Costa+Higuchi.pdf.txtbca323840bef5abd3255a80f743bc7e1MD53ORIGINALDisserta??o_Kiyo Costa Higuchi.pdfDisserta??o_Kiyo Costa Higuchi.pdfapplication/pdf4067661http://tede2.pucrs.br/tede2/bitstream/tede/10070/2/Disserta%C3%A7%C3%A3o_Kiyo+Costa+Higuchi.pdf352d8f6c66669a1586d4da3b642fd872MD52LICENSElicense.txtlicense.txttext/plain; charset=utf-8590http://tede2.pucrs.br/tede2/bitstream/tede/10070/1/license.txt220e11f2d3ba5354f917c7035aadef24MD51tede/100702022-02-02 12:00:20.857oai:tede2.pucrs.br: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Biblioteca Digital de Teses e Dissertaçõeshttp://tede2.pucrs.br/tede2/PRIhttps://tede2.pucrs.br/oai/requestbiblioteca.central@pucrs.br||opendoar:2022-02-02T14:00:20Biblioteca Digital de Teses e Dissertações da PUC_RS - Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS)false |
dc.title.por.fl_str_mv |
Urease de helicobacter pylori : produ??o, marca??o fluorescente e estudo da biodistribui??o por imageamento ?ptico in vivo |
title |
Urease de helicobacter pylori : produ??o, marca??o fluorescente e estudo da biodistribui??o por imageamento ?ptico in vivo |
spellingShingle |
Urease de helicobacter pylori : produ??o, marca??o fluorescente e estudo da biodistribui??o por imageamento ?ptico in vivo Higuchi, Kiyo Costa Urease de Helicobacter Pylori Imageamento ?ptico in Vivo Biodistribui??o Fluor?foro Biodistribution Fluoroprobe Helicobacter Pylori Urease In Vivo Optical Imaging ENGENHARIAS |
title_short |
Urease de helicobacter pylori : produ??o, marca??o fluorescente e estudo da biodistribui??o por imageamento ?ptico in vivo |
title_full |
Urease de helicobacter pylori : produ??o, marca??o fluorescente e estudo da biodistribui??o por imageamento ?ptico in vivo |
title_fullStr |
Urease de helicobacter pylori : produ??o, marca??o fluorescente e estudo da biodistribui??o por imageamento ?ptico in vivo |
title_full_unstemmed |
Urease de helicobacter pylori : produ??o, marca??o fluorescente e estudo da biodistribui??o por imageamento ?ptico in vivo |
title_sort |
Urease de helicobacter pylori : produ??o, marca??o fluorescente e estudo da biodistribui??o por imageamento ?ptico in vivo |
author |
Higuchi, Kiyo Costa |
author_facet |
Higuchi, Kiyo Costa |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Papal?o, Ricardo Meurer |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/1933730859000512 |
dc.contributor.advisor-co1.fl_str_mv |
Carlini, C?lia Regina Ribeiro da Silva |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/5587901760491970 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/8980765616740339 |
dc.contributor.author.fl_str_mv |
Higuchi, Kiyo Costa |
contributor_str_mv |
Papal?o, Ricardo Meurer Carlini, C?lia Regina Ribeiro da Silva |
dc.subject.por.fl_str_mv |
Urease de Helicobacter Pylori Imageamento ?ptico in Vivo Biodistribui??o Fluor?foro Biodistribution Fluoroprobe |
topic |
Urease de Helicobacter Pylori Imageamento ?ptico in Vivo Biodistribui??o Fluor?foro Biodistribution Fluoroprobe Helicobacter Pylori Urease In Vivo Optical Imaging ENGENHARIAS |
dc.subject.eng.fl_str_mv |
Helicobacter Pylori Urease In Vivo Optical Imaging |
dc.subject.cnpq.fl_str_mv |
ENGENHARIAS |
description |
Alzheimer?s Disease (AD) has the presence of brain neurofibrillary tangles ? deposits of the hyperphosphorylated tau protein ? among its histopathological hallmarks. AD patients are more often infected with Helicobacter pylori (a gastric pathogen) than healthy individuals, and it has been reported that an H. pylori culture filtrate induced tau hyperphosphorylation in vitro and in vivo. H. pylori infects about half of the world?s population, and its survival in the human stomach is only viable due to the enzyme urease (H. pylori urease, HPU), possibly involved in AD pathogenesis, although little is known about its in vivo biodistribution. The present work aimed to investigate HPU?s absorption and biodistribution kinetics in the nude mouse (Mus musculus Foxn1nu) model using in vivo optical imaging. For this purpose, HPU was produced, characterized, labeled with fluorophores, and given intraperitoneally to the animals. The protein?s oligomerization states were monitored during weeks, and it displayed stability in the dodecameric form. In silico studies indicated the presence of potentially free amino acid residues in HPU?s surface for fluorescent labeling, and the latter did not affect HPU?s enzymatic activity. The developed methodology was efficient in acquiring in vivo images to monitor the biodistribution and absorption and excretion kinetics of the labeled HPU. Rapid absorption of the protein by the circulatory system from the peritoneal cavity and later excretion by the kidneys was observed, followed by the migration to the bladder ? approximately 40 min after the injection. Further investigations involving chronic treatment with HPU are required to seek deeper clarification on the mechanisms through which the enzyme, produced by a gastric pathogen, may be involved in neuropathogenic processes. |
publishDate |
2021 |
dc.date.issued.fl_str_mv |
2021-08-30 |
dc.date.accessioned.fl_str_mv |
2022-02-01T18:30:21Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
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http://tede2.pucrs.br/tede2/handle/tede/10070 |
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http://tede2.pucrs.br/tede2/handle/tede/10070 |
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por |
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por |
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4953914605093919665 |
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500 500 600 |
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4518971056484826825 |
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3590462550136975366 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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Pontif?cia Universidade Cat?lica do Rio Grande do Sul |
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Programa de P?s-Gradua??o em Engenharia e Tecnologia de Materiais |
dc.publisher.initials.fl_str_mv |
PUCRS |
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Brasil |
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Escola Polit?cnica |
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Pontif?cia Universidade Cat?lica do Rio Grande do Sul |
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