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1.
Pediatr Allergy Immunol ; 32(6): 1296-1306, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-33876465

RESUMO

BACKGROUND: Anaphylaxis is the most severe manifestation of allergic disorders. The poor knowledge of its molecular mechanisms often leads to under-diagnosis. MicroRNAs (miRNA) regulate physiologic and pathologic processes, and they have been postulated as promising diagnostic markers. The main objectives of this study were to characterize the human miRNA profile during anaphylaxis and to assess their capacity as diagnostic markers and determine their participation in the molecular mechanisms of this event. METHODS: The miRNA serum profiles from the acute and baseline phase of 5 oral food-challenged anaphylactic children (<18 years old) were obtained by next-generation sequencing (NGS). From the panel of statistically significant miRNAs obtained, several candidates were selected and analyzed in 19 anaphylactic children by qPCR. We performed system biology analysis (SBA) on their target genes to identify main functions and canonical pathways. A functional in vitro assay was carried out incubating endothelial cells (ECs) in anaphylactic conditions. RESULTS: The NGS identified 389 miRNAs among which 41 were significantly different between acute and baseline samples. The high levels of miR-21-3p (fold change = 2.28, P = .006) and miR-487b-3p (fold change = 1.04, P = .039) observed by NGS in acute serum samples were confirmed in a larger group of 19 patients. The SBA revealed molecular pathways related to the inflammation and immune system regulation. miR-21-3p increased intracellularly and in acute phase serum after EC stimulation. CONCLUSIONS: These findings provide, for the first time, some insights into the anaphylactic miRNA serum profile in children and point to miR-21-3p and miR-487b-3p as candidate biomarkers. Furthermore, the SBA revealed a possible implication of these molecules in the underlying molecular mechanisms. Moreover, ECs increased miR-21-3p intracellularly and released it to the environment in response to anaphylaxis.


Assuntos
Anafilaxia , MicroRNAs , Adolescente , Biomarcadores , Criança , Células Endoteliais , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos
3.
Ginecol Obstet Mex ; 83(10): 614-26, 2015 Oct.
Artigo em Espanhol | MEDLINE | ID: mdl-26859923

RESUMO

Polycystic ovary syndrome (PCOS) is known as a common gynecologic and endocrinology disease with multiple short and long-term consequences. It is one of the most common causes for hyperandrogenism and anovulation, increases the risk for metabolic syndrome, type 2 diabetes and cardiovascular disease. Its etiology remains unclear. PCOS is thought to be the result of the interaction between predisposing genetic variants with environmental factors and strongly depends on ethnicity. Proteomics allows the study of several hundreds or thousands of proteins in order to reveal physiological state of a tissue or an organ at the molecular level and to identify disease-specific biomarkers. Its use on PCOS patients will permit us to identify molecules that are involved in the PCOS pathology so we can develop specific diagnostic and management approaches.


Assuntos
Biomarcadores/metabolismo , Síndrome do Ovário Policístico/fisiopatologia , Proteômica/métodos , Feminino , Predisposição Genética para Doença , Humanos , Síndrome do Ovário Policístico/complicações , Síndrome do Ovário Policístico/etiologia
4.
J Proteomics ; 95: 84-8, 2013 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-23500130

RESUMO

Mass spectrometry is already a well-established protein identification tool and recent methodological and technological developments have also made possible the extraction of quantitative data of protein abundance in large-scale studies. Several strategies for absolute and relative quantitative proteomics and the statistical assessment of quantifications are possible, each having specific measurements and therefore, different data analysis workflows. The guidelines for Mass Spectrometry Quantification allow the description of a wide range of quantitative approaches, including labeled and label-free techniques and also targeted approaches such as Selected Reaction Monitoring (SRM). BIOLOGICAL SIGNIFICANCE: The HUPO Proteomics Standards Initiative (HUPO-PSI) has invested considerable efforts to improve the standardization of proteomics data handling, representation and sharing through the development of data standards, reporting guidelines, controlled vocabularies and tooling. In this manuscript, we describe a key output from the HUPO-PSI-namely the MIAPE Quant guidelines, which have developed in parallel with the corresponding data exchange format mzQuantML [1]. The MIAPE Quant guidelines describe the HUPO-PSI proposal concerning the minimum information to be reported when a quantitative data set, derived from mass spectrometry (MS), is submitted to a database or as supplementary information to a journal. The guidelines have been developed with input from a broad spectrum of stakeholders in the proteomics field to represent a true consensus view of the most important data types and metadata, required for a quantitative experiment to be analyzed critically or a data analysis pipeline to be reproduced. It is anticipated that they will influence or be directly adopted as part of journal guidelines for publication and by public proteomics databases and thus may have an impact on proteomics laboratories across the world. This article is part of a Special Issue entitled: Standardization and Quality Control.


Assuntos
Espectrometria de Massas/normas , Proteômica/normas , Guias como Assunto , Espectrometria de Massas/métodos , Proteômica/métodos
5.
J Proteomics ; 73(7): 1404-9, 2010 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-20167299

RESUMO

We have set up a fast and easy methodology to identify cell-surface proteins in live yeasts. A non-gel proteomic approach was based on a short period of trypsin treatment followed by peptide separation and identification using nano-LC followed by off-line MS/MS. Candida albicans was used as a model organism and proteins involved in cell wall organization, cell rescue, defense, virulence, transport, protein fate and metabolism were identified. This strategy is a powerful tool to study host-pathogen interactions and to look for potential vaccine candidates and drug targets.


Assuntos
Candida albicans/metabolismo , Membrana Celular/metabolismo , Sistema Livre de Células/química , Cromatografia Líquida/métodos , Proteínas Fúngicas/análise , Espectrometria de Massas/métodos , Proteoma/análise
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