Development and validation of an assay for measurement of leptin in pig saliva
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1186/s12917-016-0871-9 http://hdl.handle.net/11449/173694 |
Resumo: | Background: Leptin has been measured in human in saliva samples. However, the low leptin concentration found in this biological fluid makes necessary the use of high sensitive methods. To the authors' knowledge, leptin has not been measured in porcine saliva. This study aimed to develop and validate a time-resolved immunofluorometric assay (TR-IFMA) for salivary leptin measurements in pigs, using a species-specific antibody, and to evaluate how salivary leptin changes with body weight, food ingestion, and in experimental models of stress and inflammation. Polyclonal antibodies were produced in rabbits immunized with recombinant porcine leptin and used to develop a sandwich TR-IFMA. Results: The method had intra-assay and inter-assay coefficients of variation lower than 10 and 16 %, respectively. The assay was accurate and the low limit of detection allowed detection of leptin in all analyzed samples. Salivary leptin concentration was positively correlated to body weight (r = 0.58, P = 0.01) and increased after food ingestion (P < 0.001) and after 24 h of applying a model of experimental inflammation by turpentine injection (P < 0.05). However, it did not significantly change after a model of acute stress consisting of a nose snare restraining. Conclusion: These results indicate that the developed assay can measure leptin in porcine saliva in a reliable way and that leptin in saliva is influenced by body weight, food ingestion and inflammation. |
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Development and validation of an assay for measurement of leptin in pig salivaInflammationLeptinPorcine salivaStressValidationBackground: Leptin has been measured in human in saliva samples. However, the low leptin concentration found in this biological fluid makes necessary the use of high sensitive methods. To the authors' knowledge, leptin has not been measured in porcine saliva. This study aimed to develop and validate a time-resolved immunofluorometric assay (TR-IFMA) for salivary leptin measurements in pigs, using a species-specific antibody, and to evaluate how salivary leptin changes with body weight, food ingestion, and in experimental models of stress and inflammation. Polyclonal antibodies were produced in rabbits immunized with recombinant porcine leptin and used to develop a sandwich TR-IFMA. Results: The method had intra-assay and inter-assay coefficients of variation lower than 10 and 16 %, respectively. The assay was accurate and the low limit of detection allowed detection of leptin in all analyzed samples. Salivary leptin concentration was positively correlated to body weight (r = 0.58, P = 0.01) and increased after food ingestion (P < 0.001) and after 24 h of applying a model of experimental inflammation by turpentine injection (P < 0.05). However, it did not significantly change after a model of acute stress consisting of a nose snare restraining. Conclusion: These results indicate that the developed assay can measure leptin in porcine saliva in a reliable way and that leptin in saliva is influenced by body weight, food ingestion and inflammation.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)São Paulo State University (FMVZ - UNESP) Department of Veterinary Clinical Sciences School of Veterinary Medicine and Animal Science, campus of Botucatu. Rua Dr. Walter Mauricio Correa, s/nUniversity of Murcia Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU) Veterinary Clinical Hospital, Campus of Excellence Mare NostrumUniversity of Murcia Animal Production Department Veterinary School, Campus of Excellence Mare NostrumUniversity Autonoma of Barcelona Department of Animal Medicine and Surgery Veterinary SchoolSão Paulo State University (FMVZ - UNESP) Department of Veterinary Clinical Sciences School of Veterinary Medicine and Animal Science, campus of Botucatu. Rua Dr. Walter Mauricio Correa, s/nCNPq: PDE 203241/2014-2Universidade Estadual Paulista (Unesp)Veterinary Clinical HospitalVeterinary SchoolSchmidt, Elizabeth M.S. [UNESP]Escribano, DamiánMartinez-Subiela, SilviaMartinez-Miró, SilviaHernández, FuensantaTvarijonaviciute, AstaCerón, José J.Tecles, Fernando2018-12-11T17:07:19Z2018-12-11T17:07:19Z2016-10-28info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://dx.doi.org/10.1186/s12917-016-0871-9BMC Veterinary Research, v. 12, n. 1, 2016.1746-6148http://hdl.handle.net/11449/17369410.1186/s12917-016-0871-92-s2.0-849926093852-s2.0-84992609385.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBMC Veterinary Research0,934info:eu-repo/semantics/openAccess2023-11-09T06:08:51Zoai:repositorio.unesp.br:11449/173694Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T17:12:48.820312Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Development and validation of an assay for measurement of leptin in pig saliva |
title |
Development and validation of an assay for measurement of leptin in pig saliva |
spellingShingle |
Development and validation of an assay for measurement of leptin in pig saliva Schmidt, Elizabeth M.S. [UNESP] Inflammation Leptin Porcine saliva Stress Validation |
title_short |
Development and validation of an assay for measurement of leptin in pig saliva |
title_full |
Development and validation of an assay for measurement of leptin in pig saliva |
title_fullStr |
Development and validation of an assay for measurement of leptin in pig saliva |
title_full_unstemmed |
Development and validation of an assay for measurement of leptin in pig saliva |
title_sort |
Development and validation of an assay for measurement of leptin in pig saliva |
author |
Schmidt, Elizabeth M.S. [UNESP] |
author_facet |
Schmidt, Elizabeth M.S. [UNESP] Escribano, Damián Martinez-Subiela, Silvia Martinez-Miró, Silvia Hernández, Fuensanta Tvarijonaviciute, Asta Cerón, José J. Tecles, Fernando |
author_role |
author |
author2 |
Escribano, Damián Martinez-Subiela, Silvia Martinez-Miró, Silvia Hernández, Fuensanta Tvarijonaviciute, Asta Cerón, José J. Tecles, Fernando |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Veterinary Clinical Hospital Veterinary School |
dc.contributor.author.fl_str_mv |
Schmidt, Elizabeth M.S. [UNESP] Escribano, Damián Martinez-Subiela, Silvia Martinez-Miró, Silvia Hernández, Fuensanta Tvarijonaviciute, Asta Cerón, José J. Tecles, Fernando |
dc.subject.por.fl_str_mv |
Inflammation Leptin Porcine saliva Stress Validation |
topic |
Inflammation Leptin Porcine saliva Stress Validation |
description |
Background: Leptin has been measured in human in saliva samples. However, the low leptin concentration found in this biological fluid makes necessary the use of high sensitive methods. To the authors' knowledge, leptin has not been measured in porcine saliva. This study aimed to develop and validate a time-resolved immunofluorometric assay (TR-IFMA) for salivary leptin measurements in pigs, using a species-specific antibody, and to evaluate how salivary leptin changes with body weight, food ingestion, and in experimental models of stress and inflammation. Polyclonal antibodies were produced in rabbits immunized with recombinant porcine leptin and used to develop a sandwich TR-IFMA. Results: The method had intra-assay and inter-assay coefficients of variation lower than 10 and 16 %, respectively. The assay was accurate and the low limit of detection allowed detection of leptin in all analyzed samples. Salivary leptin concentration was positively correlated to body weight (r = 0.58, P = 0.01) and increased after food ingestion (P < 0.001) and after 24 h of applying a model of experimental inflammation by turpentine injection (P < 0.05). However, it did not significantly change after a model of acute stress consisting of a nose snare restraining. Conclusion: These results indicate that the developed assay can measure leptin in porcine saliva in a reliable way and that leptin in saliva is influenced by body weight, food ingestion and inflammation. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-10-28 2018-12-11T17:07:19Z 2018-12-11T17:07:19Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1186/s12917-016-0871-9 BMC Veterinary Research, v. 12, n. 1, 2016. 1746-6148 http://hdl.handle.net/11449/173694 10.1186/s12917-016-0871-9 2-s2.0-84992609385 2-s2.0-84992609385.pdf |
url |
http://dx.doi.org/10.1186/s12917-016-0871-9 http://hdl.handle.net/11449/173694 |
identifier_str_mv |
BMC Veterinary Research, v. 12, n. 1, 2016. 1746-6148 10.1186/s12917-016-0871-9 2-s2.0-84992609385 2-s2.0-84992609385.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
BMC Veterinary Research 0,934 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
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1808128773328994304 |