40 resultados para BIOLOGICAL SAMPLES
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Pós-graduação em Medicina Veterinária - FMVZ
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The complexity of biological samples poses a major challenge for reliable compound identification in mass spectrometry (MS). The presence of interfering compounds that cause additional peaks in the spectrum can make interpretation and assignment difficult. To overcome this issue, new approaches are needed to reduce complexity and simplify spectral interpretation. Recently, focused on unknown metabolite identification, we presented a new approach, RANSY (ratio analysis of nuclear magnetic resonance spectroscopy; Anal. Chem. 2011, 83, 7616-7623), which extracts the signals related to the same metabolite based on peak intensity ratios. On the basis of this concept, we present the ratio analysis of mass spectrometry (RAMSY) method, which facilitates improved compound identification in complex MS spectra. RAMSY works on the principle that, under a given set of experimental conditions, the abundance/intensity ratios between the mass fragments from the same metabolite are relatively constant. Therefore, the quotients of average peak ratios and their standard deviations, generated using a small set of MS spectra from the same ion chromatogram, efficiently allow the statistical recovery of the metabolite peaks and facilitate reliable identification. RAMSY was applied to both gas chromatography/MS and liquid chromatography tandem MS (LC-MS/MS) data to demonstrate its utility. The performance of RAMSY is typically better than the results from correlation methods. RAMSY promises to improve unknown metabolite identification for MS users in metabolomics or other fields.
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Melanin is a very important pigment to human species, and besides defining skin, eyes and hair color, it is also involved in sun exposure protection. This pigment is classified into two subtypes: pheomelanin, which is responsible for lighter pigmentation and eumelanin, the dark pigment related. Due to this type of melanin variation it is possible to found different phenotypes of hair and skin color. The genetics MC1R variations are described as the most important for diversity in pigmentation, and this gene is directly related to the complex process of melanin synthesis. The use of these variations to phenotype prediction using genetic information has been used in Europe countries to infer physical features from biological samples, with the purpose of directing searches of criminal suspects and victims identification. The aim of this study was to analyze 8 major MC1R polymorphisms in a sample of Brazilian individuals. Analyzing 91 individuals, we observed with higher frequencies the polymorphisms rs1805005:G>T, rs2228479:G>A and rs885479:G>A, which are related to skin and hair colors, as previously showed in literature. These data suggest the possibility of predicting phenotype from genetic polymorphisms in Brazilian population. However it will be necessary to analyze a larger number of individuals to be able to confirm these associations and to perform a more detailed statistical analysis
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To determinate the multielemental concentrations in biological samples using the technique of total reflection X-ray fluorescence (TRFX) is essential the use of standard samples. The aim of this study is the determination of the sensitivities of the relevant elements in standard samples (Multi Element atomic spectroscopy standard solution in HNO3 I) prepared in three solutions with different and known concentrations. Based on the sensitivities of the elements contained in the multilelementar standard sample, it can estimate the sensitivity for an element found in the biological sample and consequently it is possible to determinate the concentration, even though the standard does not contain this element. The measurement of fluorescence was performed at the Brazilian Synchrotron Light Laboratory (LNLS) in Campinas
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(Microarray technology in study of head neck cancer). The microarray technology is a tool for global analysis of gene expression that allows investigating hundreds or thousands of genes in a sample using a hybridization reaction. This technology is based on hybridization between labeled targets derived from biological samples and an array of many DNA probes immobilized on a solid matrix, representing the genes of interest. The simultaneous study of hundreds of genes became the microarray technique a very important tool of global analysis, with applications in several areas, including the study of the development of cancer. Head and neck squamous cell carcinoma (HNSCC) is the fifth most common cancer worldwide, with a global annual incidence of 780,000 new cases. Large-scale studies involving microarrays have identified specific gene expression signatures associated with expression changes in HNSCC samples compared to normal tissue, as well as genes involved in clinical outcome and metastasis. However, the considerable heterogeneity among these studies occurs due to experimental design, number of samples, disease sites and stage, choice of microarray platform and results validation. Thus, there is much to be validated, before the technique has clinical utility. In relation to head and neck neoplasia, the large-scale gene analysis is very important, since the clinical and histopathological methods currently used appear to be insufficient to predict clinical progression and response to treatment. Thus, this approach could result in more effective diagnostic and prognostic and most appropriate therapy for this neoplasia.
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
A critical review of the properties of fusidic acid and the analytical methods for its determination
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Fusidic acid, an antibiotic produced from the Fusidium coccineum fungus, belongs to the class of steroids, but has no corticosteroid effects. It is indicated for the treatment of infections caused by methicillin-resistant Staphylococcus aureus strains. The aim of this study was to search for the properties of fusidic acid published so far in the literature, as well as the methods developed for its determination in biological samples and pharmaceutical formulations. From the findings, we can conclude that fusidic acid has been used for decades and is indicated for the treatment of serious infections caused by Gram-positive microorganisms to this day. Furthermore, it is a hypoallergenic agent, has low toxicity, shows low resistance, and has no cross-resistance with other clinically used antibiotics. The analytical method of high-performance liquid chromatography has been widely used, since it can reduce the cost and time of analysis, making it more viable for routine quality control in the pharmaceutical industry.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Background: Plasmodium vivax circumsporozoite variants have been identified in several geographical areas. The real implication of the genetic variation in this region of the P. vivax genome has been questioned for a long time. Although previous studies have observed significant association between VK210 and the Duffy blood group, we present here that evidences of this variation are limited to the CSP central portion.Methods: The phylogenetic analyses were accomplished starting from the amplification of conserved domains of 18 SSU RNAr and Cyt B. The antibodies responses against the CSP peptides, MSP-1, AMA-1 and DBP were detected by ELISA, in plasma samples of individuals infected with two P. vivax CS genotypes: VK210 and P. vivax-like.Results: These analyses of the two markers demonstrate high similarity among the P. vivax CS genotypes and surprisingly showed diversity equal to zero between VK210 and P. vivax-like, positioning these CS genotypes in the same clade. A high frequency IgG antibody against the N- and C-terminal regions of the P. vivax CSP was found as compared to the immune response to the R- and V-repetitive regions (p = 0.0005, Fisher's Exact test). This difference was more pronounced when the P. vivax-like variant was present in the infection (p = 0.003, Fisher's Exact test). A high frequency of antibody response against MSP-1 and AMA-1 peptides was observed for all P. vivax CS genotypes in comparison to the same frequency for DBP.Conclusions: This results target that the differences among the P. vivax CS variants are restrict to the central repeated region of the protein, mostly nucleotide variation with important serological consequences.
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Neuston samples collected from the Charleston Bump region off the coast of South Carolina, U.S.A., during the summers of 2002 and 2003 consistently included a decapod species of undetermined identity with a large brachyuran megalopa. Despite their resemblance to some calappids, it was impossible to make a definitive identification based solely on general morphology. Therefore, additional neuston tows were taken on the continental shelf near Charleston, during the summer of 2004 to obtain these living megalopae. These were raised successfully through five juvenile stages at the Southeastern Regional Taxonomic Center (SERTC) laboratory. The morphology of the juveniles provided evidence that they are megalopae of Calappa tortugae Rathbun, 1933. Comparisons with megalopae of Hepatus epheliticus (Linnaeus, 1763), H. pudibundus (Herbst, 1785), Calappa flammea (Herbst, 1794) and Cryptosoma balguerii (Desbonne, 1867) are presented here. This is the first complete description of the megalopa morphology of a member of the genus Calappa Weber, 1795 from the Western Atlantic, and it is helpful for taxonomic, systematic and ecological purposes.