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      A chemometric model applied to fatty acid determination in blood

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          Abstract

          Changes in fatty acid profiles have been associated with several pathophysiological processes. Gas chromatography-mass spectrometry (GC-MS) data monitoring of selected ions was used with principal components analysis (PCA), revealing a set of relevant ions for quantification and characterization of fatty acids. This protocol was successfully applied to the analyses of fatty acids in different human blood lipids, allowing the quantification of several fatty acids and revealing their unsaturation numbers. Moreover the presences of contaminants, artifacts and co-eluitions in the chromatogram were also revealed without any additional analyses. Thus, fatty acid constituents of triglycerides, phospholipids and esters of cholesterol present in plasma and erythrocyte membranes were accurately determined, with repeatability, low limits of determination and of quantification.

          Translated abstract

          Mudanças no perfil de ácidos graxos têm sido associadas a vários processos patofisiológicos. Dados obtidos por cromatografia gasosa-espectrometria de massas (GC-MS) usando monitoramento seletivo de íons foram empregados para revelarem, após análise por análise de componentes principais (PCA), um conjunto relevante de íons para quantificação e caracterização de ácidos graxos. Este protocolo foi aplicado com sucesso na análise de ácidos graxos presentes em diversas frações lipídicas de sangue, permitindo a quantificação de diversos ácidos graxos e revelando seus números de insaturações. Adicionalmente, a presença de contaminantes, artefatos e co-eluições no cromatograma também foram reveladas sem análises adicionais. Por fim, ácidos graxos presentes em triacilglicerídeos, fosfolipídios e ésteres de colesterol presentes em plasma e membrana de eritrócitos foram determinados com exatidão adequada, repetibilidade e baixos limites de detecção e de quantificação.

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          Most cited references27

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          Functional indices of vitamin D status and ramifications of vitamin D deficiency.

          Serum 25-hydroxyvitamin D3 [25(OH)D3] concentrations are currently recognized as the functional status indicator for vitamin D. Evidence is reviewed that shows that serum 25(OH)D3 concentrations of < 80 nmol/L are associated with reduced calcium absorption, osteoporosis, and increased fracture risk. For typical older individuals, supplemental oral intakes of approximately 1300 IU/d are required to reach the lower end of the optimal range. Evidence of substantial problems in routine clinical measurement of serum 25(OH)D3 concentrations among patients is cited. There is great need for standardization and improved reproducibility and sensitivity of measurements of serum 25(OH)D3 concentrations.
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            A highly conserved binding site in vesicle-associated membrane protein-associated protein (VAP) for the FFAT motif of lipid-binding proteins.

            A variety of lipid-binding proteins contain a recently described motif, designated FFAT (two phenylalanines in an acidic tract), which binds to vesicle-associated-membrane protein-associated protein (VAP). VAP is a conserved integral membrane protein of the endoplasmic reticulum that contains at its amino terminus a domain related to the major sperm protein of nematode worms. Here we have studied the FFAT-VAP interaction in Saccharomyces cerevisiae, where the VAP homologue Scs2 regulates phospholipid metabolism via an interaction with the FFAT motif of Opi1. By introducing mutations at random into Scs2, we found that mutations that abrogated binding to FFAT were clustered in the most highly conserved region. Using site-directed mutagenesis, we identified several critical residues, including two lysines widely separated in the primary sequence. By examining all other conserved basic residues, we identified a third residue that was moderately important for binding FFAT. Modeling VAP on the known structure of major sperm protein showed that the critical residues form a patch on a positively charged face of the protein. In vivo functional studies of SCS22, a second SCS2-like gene in S. cerevisiae, showed that SCS2 was the dominant gene in the regulation of Opi1, with a minor contribution from SCS22. We then established that reduction in the affinity of Scs2 mutants for FFAT correlated well with loss of function, indicating the importance of these residues for binding FFAT motifs. Finally, we found that human VAP-A could substitute for Scs2 but that it functioned poorly, suggesting that other factors modulate the binding of Scs2 to proteins with FFAT motifs.
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              Regulation of milk protein gene expression.

              Studies using both transgenic mice and transfected mammary epithelial cells have established that composite response elements containing multiple binding sites for several transcription factors mediate the hormonal and developmental regulation of milk protein gene expression. Activation of signal transduction pathways by lactogenic hormones and cell-substratum interactions activate transcription factors and change chromatin structure and milk protein gene expression. The casein promoters have binding sites for signal transducers and activators of transcription 5, Yin Yang 1, CCAAT/enhancer binding protein, and the glucocorticoid receptor. The whey protein gene promoters have binding sites for nuclear factor I, as well as the glucocorticoid receptor and the signal transducers and activators of transcription 5. The functional importance of some of these factors in mammary gland development and milk protein gene expression has been elucidated by studying mice in which some of these factors have been deleted.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                jbchs
                Journal of the Brazilian Chemical Society
                J. Braz. Chem. Soc.
                Sociedade Brasileira de Química (São Paulo )
                1678-4790
                October 2013
                : 24
                : 10
                : 1599-1605
                Affiliations
                [1 ] Universidade Estadual de Campinas Brazil
                [2 ] Universidade Estadual de Campinas Brazil
                Article
                S0103-50532013001000007
                10.5935/0103-5053.20130201
                93eb9e0c-a20d-4302-b14d-64fbdb1236ad

                http://creativecommons.org/licenses/by/4.0/

                History
                Product

                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=0103-5053&lng=en
                Categories
                CHEMISTRY, MULTIDISCIPLINARY

                General chemistry
                principal component analysis,selected ion monitoring,fatty acid methyl esters

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