Impactos da carbamilação e da icterícia induzidas in vitro sobre a quantificação de parâmetros laboratoriais associados à coagulação sanguínea

Detalhes bibliográficos
Autor(a) principal: Mastella, Aline Klein
Data de Publicação: 2023
Tipo de documento: Tese
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
dARK ID: ark:/26339/001300000rw3b
Texto Completo: http://repositorio.ufsm.br/handle/1/27840
Resumo: Hemostasis is a process highly regulated by pro-coagulant and anticoagulant proteins with the purpose of maintaining blood flow within the vessels and capable of promoting coagulation when there is vascular damage. Changes in different stages of hemostasis predispose to diseases that lead to hemorrhage, as in hemophilia, or to thrombotic events and can be diagnosed through the analysis of parameters that evaluate the individual's coagulation. For the results from the coagulation tests to be carried out correctly, error factors must be avoided. Interferents such as lipemia, icterus and hemolysis are possible sources that can generate results that are incompatible with reality because they cause changes in the analysis. Another factor that can affect the result of laboratory tests is the structural alteration of coagulation proteins. One of these changes comes from the carbamylation reaction that occurs in amino acids, peptides and proteins, affecting its function and predisposing to some pathologies. Carbamylation occurs in the body mainly due to the presence of cyanate due to urea metabolism and the action of myeloperoxidase in inflammatory processes on thiocyanate and can affect proteins involved in hemostasis. Although some studies have already been developed, research that better evaluates the possible interference of icterus in the determination of D-dimer and protein carbamylation in the evaluation of coagulation parameters are still necessary for a better evaluation of the patient and understanding of interferences in hemostasis. In this first study, the interference of icterus in the D-dimer analysis was evaluated through the addition of commercial D-dimer control and different concentrations of bilirubin (0, 0.9, 1.9, 3.8, 7.5, 15 and 30 mg/dL) to human plasma pools, followed by D-dimer quantification by immunoturbidimetric assay. Carbamylation was analyzed by exposing comercial controls and plasma pools to different concentrations of potassium cyanate (0, 150 nm, 150 μM, and 150 mM) and urea (0, 20, 100, and 500 md/dL) with subsequent analysis of coagulation parameters: prothrombin time (PT), activated partial thromboplastin time (aTTP), and fibrinogen. Through the studies performed, it can be verified that icterus does not seem to interfere in the analysis of D-dimer. The carbamylation experiments with KOCN showed a significant increase in TP and aTTP and reduction in fibrinogen in the commercial controls, while in plasma the changes in the tests occurred at the highest concentration of cyanate, except for TP, which also elevated at the 150 μM KOCN concentration. Incubation with increasing concentrations of urea prolonged the PT and aTTP in the commercial controls. In plasma, such evidence occurred only at the highest urea concentration. Fibrinogen was not shown to be affected by incubation with urea. The observed changes may indicate that patients with inflammatory processes and chronic kidney disease could develop hemostatic disorders.
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spelling Impactos da carbamilação e da icterícia induzidas in vitro sobre a quantificação de parâmetros laboratoriais associados à coagulação sanguíneaImpacts of in vitro-induced carbamilation and icterus on the quantification of laboratory parameters associated with blood coagulationHemostasiaInterferentesCarbamilaçãoParâmetros da coagulaçãoHemostasisInterferentsCarbamylationCoagulation parametersCNPQ::CIENCIAS DA SAUDE::FARMACIAHemostasis is a process highly regulated by pro-coagulant and anticoagulant proteins with the purpose of maintaining blood flow within the vessels and capable of promoting coagulation when there is vascular damage. Changes in different stages of hemostasis predispose to diseases that lead to hemorrhage, as in hemophilia, or to thrombotic events and can be diagnosed through the analysis of parameters that evaluate the individual's coagulation. For the results from the coagulation tests to be carried out correctly, error factors must be avoided. Interferents such as lipemia, icterus and hemolysis are possible sources that can generate results that are incompatible with reality because they cause changes in the analysis. Another factor that can affect the result of laboratory tests is the structural alteration of coagulation proteins. One of these changes comes from the carbamylation reaction that occurs in amino acids, peptides and proteins, affecting its function and predisposing to some pathologies. Carbamylation occurs in the body mainly due to the presence of cyanate due to urea metabolism and the action of myeloperoxidase in inflammatory processes on thiocyanate and can affect proteins involved in hemostasis. Although some studies have already been developed, research that better evaluates the possible interference of icterus in the determination of D-dimer and protein carbamylation in the evaluation of coagulation parameters are still necessary for a better evaluation of the patient and understanding of interferences in hemostasis. In this first study, the interference of icterus in the D-dimer analysis was evaluated through the addition of commercial D-dimer control and different concentrations of bilirubin (0, 0.9, 1.9, 3.8, 7.5, 15 and 30 mg/dL) to human plasma pools, followed by D-dimer quantification by immunoturbidimetric assay. Carbamylation was analyzed by exposing comercial controls and plasma pools to different concentrations of potassium cyanate (0, 150 nm, 150 μM, and 150 mM) and urea (0, 20, 100, and 500 md/dL) with subsequent analysis of coagulation parameters: prothrombin time (PT), activated partial thromboplastin time (aTTP), and fibrinogen. Through the studies performed, it can be verified that icterus does not seem to interfere in the analysis of D-dimer. The carbamylation experiments with KOCN showed a significant increase in TP and aTTP and reduction in fibrinogen in the commercial controls, while in plasma the changes in the tests occurred at the highest concentration of cyanate, except for TP, which also elevated at the 150 μM KOCN concentration. Incubation with increasing concentrations of urea prolonged the PT and aTTP in the commercial controls. In plasma, such evidence occurred only at the highest urea concentration. Fibrinogen was not shown to be affected by incubation with urea. The observed changes may indicate that patients with inflammatory processes and chronic kidney disease could develop hemostatic disorders.A hemostasia é um processo altamente regulado por proteínas pró-coagulantes e anticoagulantes com a finalidade de manter a fluidez sanguínea dentro dos vasos e capaz de promover a coagulação quando há dano vascular. Alterações em diferentes etapas da hemostasia predispõem a doenças que levam a hemorragias, como nas hemofilias, ou a eventos trombóticos e podem ser diagnosticadas através da análise de parâmetros que avaliam a coagulação do indivíduo. Para que os resultados provenientes dos testes de coagulação sejam realizados de forma correta, fatores de erro devem ser evitados. Interferentes como lipemia, icterícia e hemólise são possíveis fontes que podem gerar resultados incompatíveis com a realidade por causarem alteração na análise. Outro fator que pode afetar o resultado de análises laboratoriais é a alteração estrutural de proteínas da coagulação. Uma dessas alterações é proveniente da reação da carbamilação que ocorre em aminoácidos, peptídeos e proteínas, afetando a sua função e predispondo a algumas patologias. A carbamilação ocorre no organismo principalmente pela presença de cianato decorrente do metabolismo da ureia e da ação da mieloperoxidase em processos inflamatórios sobre o tiocianato e pode afetar proteínas envolvidas na hemostasia. Apesar de alguns estudos já terem sido desenvolvidos, pesquisas que avaliem melhor a possível interferência da icterícia na determinação de D-dímero e da carbamilação de proteínas na avaliação de parâmetros da coagulação ainda são necessários para uma melhor avaliação do paciente e entendimento de interferentes na hemostasia. Neste primeiro estudo, foi avaliada a interferência da icterícia na análise de D-dímero através da adição de controle comercial de D-dímero e de diferentes concentrações de bilirrubina (0, 0,9, 1,9, 3,8, 7,5, 15 e 30 mg/dL) a amostras de pool de plasma humano, seguida da quantificação do D-dímero por ensaio imunoturbidimétrico. A carbamilação, por sua vez, foi analisada através da exposição de controles da hemostasia e pools de plasma a diferentes concentrações de cianato de potássio (0, 150 nm, 150 μM e 150 mM) e ureia (0, 20, 100 e 500 md/dL) com posterior análise dos parâmetros da coagulação: tempo de protrombina (TP), tempo de tromboplastina parcial ativado (TTPa) e dosagem de fibrinogênio. Através dos estudos realizados pode-se verificar que a icterícia não parece interferir na análise de D-dímero. Os experimentos de carbamilação com KOCN demonstraram aumento significativo do TP e TTPa e redução do fibrinogênio nos controles comerciais, enquanto que no plasma as alterações nos testes ocorreram na concentração mais elevada de cianato, exceto para o TP, que também elevou na concentração de 150 μM KOCN. A incubação com concentrações crescentes de ureia prolongou o TP e o TTPa nos controles comerciais. No plasma, tal evidência só ocorreu na concentração mais elevada de ureia. O fibrinogênio não mostrou ser afetado pela incubação com ureia. As alterações observadas podem indicar que pacientes com processos inflamatórios e doença renal crônica poderiam desenvolver desordens hemostáticas.Universidade Federal de Santa MariaBrasilAnálises Clínicas e ToxicológicasUFSMPrograma de Pós-Graduação em Ciências FarmacêuticasCentro de Ciências da SaúdeMoresco, Rafael Noalhttp://lattes.cnpq.br/2269922709577261Bochi, Guilherme VargasSilva, José Edson Paz daPaniz, ClóvisComim, Fabio VasconcellosFarias, Mariela GraneroMastella, Aline Klein2023-02-17T14:30:30Z2023-02-17T14:30:30Z2023-01-17info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/27840ark:/26339/001300000rw3bporAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2023-02-17T14:30:31Zoai:repositorio.ufsm.br:1/27840Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2023-02-17T14:30:31Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Impactos da carbamilação e da icterícia induzidas in vitro sobre a quantificação de parâmetros laboratoriais associados à coagulação sanguínea
Impacts of in vitro-induced carbamilation and icterus on the quantification of laboratory parameters associated with blood coagulation
title Impactos da carbamilação e da icterícia induzidas in vitro sobre a quantificação de parâmetros laboratoriais associados à coagulação sanguínea
spellingShingle Impactos da carbamilação e da icterícia induzidas in vitro sobre a quantificação de parâmetros laboratoriais associados à coagulação sanguínea
Mastella, Aline Klein
Hemostasia
Interferentes
Carbamilação
Parâmetros da coagulação
Hemostasis
Interferents
Carbamylation
Coagulation parameters
CNPQ::CIENCIAS DA SAUDE::FARMACIA
title_short Impactos da carbamilação e da icterícia induzidas in vitro sobre a quantificação de parâmetros laboratoriais associados à coagulação sanguínea
title_full Impactos da carbamilação e da icterícia induzidas in vitro sobre a quantificação de parâmetros laboratoriais associados à coagulação sanguínea
title_fullStr Impactos da carbamilação e da icterícia induzidas in vitro sobre a quantificação de parâmetros laboratoriais associados à coagulação sanguínea
title_full_unstemmed Impactos da carbamilação e da icterícia induzidas in vitro sobre a quantificação de parâmetros laboratoriais associados à coagulação sanguínea
title_sort Impactos da carbamilação e da icterícia induzidas in vitro sobre a quantificação de parâmetros laboratoriais associados à coagulação sanguínea
author Mastella, Aline Klein
author_facet Mastella, Aline Klein
author_role author
dc.contributor.none.fl_str_mv Moresco, Rafael Noal
http://lattes.cnpq.br/2269922709577261
Bochi, Guilherme Vargas
Silva, José Edson Paz da
Paniz, Clóvis
Comim, Fabio Vasconcellos
Farias, Mariela Granero
dc.contributor.author.fl_str_mv Mastella, Aline Klein
dc.subject.por.fl_str_mv Hemostasia
Interferentes
Carbamilação
Parâmetros da coagulação
Hemostasis
Interferents
Carbamylation
Coagulation parameters
CNPQ::CIENCIAS DA SAUDE::FARMACIA
topic Hemostasia
Interferentes
Carbamilação
Parâmetros da coagulação
Hemostasis
Interferents
Carbamylation
Coagulation parameters
CNPQ::CIENCIAS DA SAUDE::FARMACIA
description Hemostasis is a process highly regulated by pro-coagulant and anticoagulant proteins with the purpose of maintaining blood flow within the vessels and capable of promoting coagulation when there is vascular damage. Changes in different stages of hemostasis predispose to diseases that lead to hemorrhage, as in hemophilia, or to thrombotic events and can be diagnosed through the analysis of parameters that evaluate the individual's coagulation. For the results from the coagulation tests to be carried out correctly, error factors must be avoided. Interferents such as lipemia, icterus and hemolysis are possible sources that can generate results that are incompatible with reality because they cause changes in the analysis. Another factor that can affect the result of laboratory tests is the structural alteration of coagulation proteins. One of these changes comes from the carbamylation reaction that occurs in amino acids, peptides and proteins, affecting its function and predisposing to some pathologies. Carbamylation occurs in the body mainly due to the presence of cyanate due to urea metabolism and the action of myeloperoxidase in inflammatory processes on thiocyanate and can affect proteins involved in hemostasis. Although some studies have already been developed, research that better evaluates the possible interference of icterus in the determination of D-dimer and protein carbamylation in the evaluation of coagulation parameters are still necessary for a better evaluation of the patient and understanding of interferences in hemostasis. In this first study, the interference of icterus in the D-dimer analysis was evaluated through the addition of commercial D-dimer control and different concentrations of bilirubin (0, 0.9, 1.9, 3.8, 7.5, 15 and 30 mg/dL) to human plasma pools, followed by D-dimer quantification by immunoturbidimetric assay. Carbamylation was analyzed by exposing comercial controls and plasma pools to different concentrations of potassium cyanate (0, 150 nm, 150 μM, and 150 mM) and urea (0, 20, 100, and 500 md/dL) with subsequent analysis of coagulation parameters: prothrombin time (PT), activated partial thromboplastin time (aTTP), and fibrinogen. Through the studies performed, it can be verified that icterus does not seem to interfere in the analysis of D-dimer. The carbamylation experiments with KOCN showed a significant increase in TP and aTTP and reduction in fibrinogen in the commercial controls, while in plasma the changes in the tests occurred at the highest concentration of cyanate, except for TP, which also elevated at the 150 μM KOCN concentration. Incubation with increasing concentrations of urea prolonged the PT and aTTP in the commercial controls. In plasma, such evidence occurred only at the highest urea concentration. Fibrinogen was not shown to be affected by incubation with urea. The observed changes may indicate that patients with inflammatory processes and chronic kidney disease could develop hemostatic disorders.
publishDate 2023
dc.date.none.fl_str_mv 2023-02-17T14:30:30Z
2023-02-17T14:30:30Z
2023-01-17
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
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url http://repositorio.ufsm.br/handle/1/27840
identifier_str_mv ark:/26339/001300000rw3b
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Análises Clínicas e Toxicológicas
UFSM
Programa de Pós-Graduação em Ciências Farmacêuticas
Centro de Ciências da Saúde
publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Análises Clínicas e Toxicológicas
UFSM
Programa de Pós-Graduação em Ciências Farmacêuticas
Centro de Ciências da Saúde
dc.source.none.fl_str_mv reponame:Manancial - Repositório Digital da UFSM
instname:Universidade Federal de Santa Maria (UFSM)
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instname_str Universidade Federal de Santa Maria (UFSM)
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institution UFSM
reponame_str Manancial - Repositório Digital da UFSM
collection Manancial - Repositório Digital da UFSM
repository.name.fl_str_mv Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)
repository.mail.fl_str_mv atendimento.sib@ufsm.br||tedebc@gmail.com
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