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5.
ERJ Open Res ; 7(1)2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33532473

RESUMO

This study found no association of the top two associated FER variants with severity of community-acquired pneumonia. Precise characterisation of phenotypes may be required in order to unravel the genetic mechanisms predisposing to poor outcome in sepsis. https://bit.ly/3jc9SmR.

8.
J Nutr Biochem ; 26(11): 1385-92, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26320676

RESUMO

Dietary intervention with ω-3 marine fatty acids may potentially modulate inflammation and oxidative stress markers related with CVD, metabolic syndrome and cancer. The aim of this study was to evaluate whether different proportions of ω-3 EPA and DHA intake provoke a modulation of the production of lipid mediators and then, an influence on different indexes of inflammation and oxidative stress in a controlled dietary animal experiment using Wistar rats. For such scope, a lipidomic SPE-LC-ESI-MS/MS approach previously developed was applied to determine lipid mediators profile in plasma samples. The effect of ω-3 fatty acids associated to different ratios EPA:DHA was compared with the effect exerted by ω-3 ALA supplementation from linseed oil and ω-6 LA from soybean oil. CRP showed a tendency to greater inflammatory status in all ω-3-fed animals. Interestingly, ratios 1:1 and 2:1 EPA:DHA evidenced a noteworthy healthy effect generating a less oxidative environment and modulating LOX and COX activities toward a decrease in the production of proinflammatory ARA eicosanoids and oxidative stress biomarkers from EPA and DHA. In addition, the ability of 1:1 and 2:1 fish oil diets to reduce lipid mediator levels was in concurrence with the protective effect exerted by decreasing inflammatory markers as ω-6/ω-3 ratio in plasma and membranes. It was also highlighted the effect of a higher DHA amount in the diet reducing the healthy benefits described in terms of inflammation and oxidative stress. Results support the antiinflammatory and antioxidative role of fish oils and, particularly, the effect of adequate proportions EPA:DHA.


Assuntos
Ácidos Docosa-Hexaenoicos/farmacologia , Ácido Eicosapentaenoico/farmacologia , Ácidos Graxos/metabolismo , Inflamação/dietoterapia , Estresse Oxidativo/efeitos dos fármacos , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Biomarcadores/metabolismo , Suplementos Nutricionais , Ácidos Docosa-Hexaenoicos/administração & dosagem , Ácido Eicosapentaenoico/administração & dosagem , Membrana Eritrocítica/efeitos dos fármacos , Membrana Eritrocítica/metabolismo , Ácidos Graxos/análise , Feminino , Óleos de Peixe/química , Óleos de Peixe/farmacologia , Inflamação/metabolismo , Óleo de Semente do Linho/química , Óleo de Semente do Linho/farmacologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Ratos Wistar , Óleo de Soja/química , Óleo de Soja/farmacologia
9.
J Mass Spectrom ; 42(11): 1391-403, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17960563

RESUMO

Mass spectrometry (MS) is a technique of paramount importance in Proteomics, and developments in this field have been possible owing to novel MS instrumentation, experimental strategies, and bioinformatics tools. Today it is possible to identify and determine relative expression levels of thousands of proteins in a biological system by MS analysis of peptides produced by proteolytic digestion. In some situations, however, the precise characterization of a particular peptide species in a very complex peptide mixture is needed. While single-fragment ion-based scanning modes such as selected ion reaction monitoring (SIRM) or consecutive reaction monitoring (CRM) may be highly sensitive, they do not produce MS/MS information and their actual specificity must be determined in advance, a prerequisite that is not usually met in a basic research context. In such cases, the MS detector may be programmed to perform continuous MS/MS spectra on the peptide ion of interest in order to obtain structural information. This selected MS/MS ion monitoring (SMIM) mode has a number of advantages that are fully exploited by MS detectors that, like the linear ion trap, are characterized by high scanning speeds. In this work, we show some applications of this technique in the context of biological studies. These results were obtained by selecting an appropriate combination of scans according to the purpose of each one of these research scenarios. They include highly specific identification of proteins present in low amounts, characterization and relative quantification of post-translational modifications such as phosphorylation and S-nitrosylation and species-specific peptide identification.


Assuntos
Fragmentos de Peptídeos/análise , Proteínas/análise , Proteômica/métodos , Espectrometria de Massas em Tandem/métodos , Animais , Membrana Celular/química , Membrana Celular/metabolismo , Conexina 43/análise , Cisteína/análogos & derivados , Cisteína/análise , Cisteína/metabolismo , Eletroforese em Gel de Poliacrilamida , Células Endoteliais/química , Células Endoteliais/metabolismo , Produtos Pesqueiros/análise , Gadiformes , Proteínas de Choque Térmico HSP90/análise , Proteínas de Choque Térmico HSP90/metabolismo , Humanos , Camundongos , Fatores de Transcrição NFATC/análise , Fatores de Transcrição NFATC/metabolismo , Fosforilação , Processamento de Proteína Pós-Traducional , Ratos , S-Nitrosotióis/análise , S-Nitrosotióis/metabolismo , Especificidade da Espécie , Tripsina/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
10.
J Proteome Res ; 6(8): 3070-80, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17622168

RESUMO

The characterization by de novo peptide sequencing of the different protein nucleoside diphosphate kinase B (NDK B) from all the commercial hakes and grenadiers belonging to the family Merlucciidae is reported. A classical proteomics approach, consisting of two-dimmensional gel electrophoresis, tryptic in-gel digestion of the excised spots, MALDI-TOF MS, LC-MS/MS, and nanoESI-MS/MS analyses, was followed for the purification and characterization of the different isoforms of the NDK B. Fragmentation spectra were used for de novo peptide sequence. A high degree of homology was found between the sequences of all the species studied and the NDK B sequence from Gillichthys mirabilis, which is accessible in the protein databases. Particular attention was paid to the differential characterization of species-specific peptides that could be used for fish authentication purposes. These findings allowed us to propose a rapid and effective classification method, based in the detection of these biomarker peptides using the selective ion reaction monitoring (SIRM) scan mode in mass spectrometry.


Assuntos
Proteínas de Peixes/análise , Peixes/metabolismo , Nucleosídeo NM23 Difosfato Quinases/análise , Peptídeos/análise , Sequência de Aminoácidos , Animais , Dados de Sequência Molecular , Nucleosídeo NM23 Difosfato Quinases/química , Especificidade da Espécie , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
11.
J Chromatogr A ; 1153(1-2): 235-58, 2007 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-17276441

RESUMO

Sample preparation is a fundamental step in the proteomics workflow. However, it is not easy to find compiled information updating this subject. In this paper, the strategies and protocols for protein extraction and identification, following either classical or second generation proteomics methodologies, are reviewed. Procedures for: tissue disruption, cell lysis, sample pre-fractionation, protein separation by 2-DE, protein digestion, mass spectrometry analysis, multidimensional peptide separations and quantification of protein expression level are described.


Assuntos
Métodos Analíticos de Preparação de Amostras/tendências , Proteínas/isolamento & purificação , Proteômica/métodos , Animais , Fracionamento Químico , Cromatografia Líquida/métodos , Eletroforese em Gel Bidimensional/métodos , Perfilação da Expressão Gênica/métodos , Humanos , Sistemas On-Line , Peptídeo Hidrolases/metabolismo , Hidrolisados de Proteína/isolamento & purificação , Kit de Reagentes para Diagnóstico , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos
12.
Sleep Breath ; 10(4): 181-7, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17053929

RESUMO

The aim of this investigation was to evaluate the contribution of cephalometry to a statistical model integrating clinical, physical, and oximetric variables, to reduce demands for polysomnographies. Two hundred and twenty-five consecutive patients that had been referred to the sleep clinic for suspected obstructive sleep apnea (OSA) were studied. The clinical assessment of all patients consisted of a sleep related questionnaire, the Epworth sleepiness scale, and a physical examination. In addition, they all underwent spirometry, cephalometry, and a full polysomnography. The clinical variables related with OSA were questions concerning witnessing of apneas by bed partners, intensity of snoring, a history of hypertension, and nocturia. A significant relation was also found with score on the Epworth scale, sex, age, body mass index, neck and waist circumferences, total number and frequency of oxygen desaturations, and the lowest oxygen saturation value. Significant cephalometric measurements were: the linear distance from gonion to gnathion, from the hyoid bone to the mandibular plane, and from the posterior nasal spine to the tip of the soft palate, and the thickness of the uvula as well. A statistical model was built to estimate a patient's probability of having OSA based on clinical variables, physical examination, pulse oximetry, and cephalometry. The validation of this model demonstrated a remarkable ability in reducing the number of polysomnographic studies. We conclude that cephalometry combined with clinical variables, physical examination, and nocturnal oximetry is useful in the diagnosis of OSA and enables the sparing of a considerable number of polysomnographies.


Assuntos
Cefalometria/estatística & dados numéricos , Polissonografia/estatística & dados numéricos , Apneia Obstrutiva do Sono/diagnóstico , Adulto , Antropometria , Índice de Massa Corporal , Estudos de Avaliação como Assunto , Feminino , Necessidades e Demandas de Serviços de Saúde/estatística & dados numéricos , Humanos , Masculino , Pessoa de Meia-Idade , Oxigênio/sangue , Exame Físico , Valor Preditivo dos Testes , Valores de Referência , Apneia Obstrutiva do Sono/psicologia , Espanha , Inquéritos e Questionários , Procedimentos Desnecessários/estatística & dados numéricos
13.
Proteomics ; 6(19): 5278-87, 2006 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16927426

RESUMO

Analysis of parvalbumin fractions through proteomic methodologies allowed the differential classification of ten commercial, closely related species of the family Merlucciidae. Muscle extracts from nine hake species of the genus Merluccius including two subspecies of Merluccius australis (australis and polylepsis) and one grenadier species Macruronus novaezelandiae with two populations (novaezelandiae and magellanicus) were evaluated by 2-DE and MALDI-TOF MS. 2-DE demonstrated that the species tested displayed a low intra-specific degree of polymorphism and the isoform patterns were noticeably species-specific. MALDI-TOF mass fingerprints showed clear differences in the pattern of peptides produced by tryptic digestion between the Merluccius and the Macruronus, making the genus differentiation possible. In addition, a selective peptide mass present in the spectra from certain hakes allowed its classification in two groups: Euro-African and American hakes. Besides, some specific masses allowed a clearly individual identification for M. bilinearis, M. australis polylepsis, M. australis australis, M. productus, M. paradoxus and M. polli, while the rest of the hake species can be grouped in two clusters, comprising M. hubbsi and M. gayi in one and M. merluccius and M. capensis in the other.


Assuntos
Proteínas de Peixes/análise , Gadiformes/metabolismo , Parvalbuminas/análise , Proteoma/análise , Sequência de Aminoácidos , Animais , Eletroforese em Gel Bidimensional , Proteínas de Peixes/química , Gadiformes/classificação , Dados de Sequência Molecular , Parvalbuminas/química , Proteoma/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
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