RESUMO
A new method for the analysis of blood lipid based on direct mass spectrometry of lipophilic low molecular weight fraction of blood plasma has been considered. Such technique allows quantification of hundreds of various types of lipids and this changes existing concepts on diagnostics of lipid disorders and related diseases. The versatility and quickness of the method significantly simplify its wide use. This method is applicable for diagnostics of atherosclerosis, diabetes, cancer and other diseases. Detalization of plasma lipid composition at the molecular level by means of mass spectrometry allows to assess the effectiveness of therapy and to optimize the drug treatment of cardiovascular diseases by phospholipid preparations.
Assuntos
Análise Química do Sangue/métodos , Lipídeos/sangue , Espectrometria de Massas/métodos , Humanos , Sensibilidade e EspecificidadeRESUMO
Application of blood plasma metabolites fingerprinting for the diagnostic of the 2nd stage of prostate cancer has been investigated. The diagnostic sensitivity (95%), specificity (96.7%) and accuracy (95.7%) of the metabolic fingerprinting were much higher then those for PSA test (35%, 83.3% and 51.7%, respectively) for the same patients. Area under the ROC-curve (0.994) suggests that the proposed approach is effective and can be used for clinical applications.
Assuntos
Biomarcadores Tumorais/sangue , Proteínas Sanguíneas/análise , Neoplasias da Próstata/sangue , Humanos , Masculino , Metaboloma , Neoplasias da Próstata/diagnóstico , Curva ROC , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
Comparative electron microscopic analysis of the patterns of centriolar and cytoplasmic microtubules immunostaining for acetylated or tyrosinated alpha-tubulin was performed during the cell cycles of the mouse diploid 3T3 (A31) and virus-transformed heteroploid SV40-3T3 cell lines. It was shown that the pattern of centriole immunostaining changed during mitosis in control 3T3 (A31) cells, but not in tumorigenic SV40-3T3 cells. A sharp change in the pattern of centriole immunostaining was observed in those cell cycle periods when the protein organization of centrosome underwent changes during interphase/mitosis or mitosis/interphase transitions due to the rearrangement of microtubule system. A high level of centriole immunostaining for tyrosinated and acetylated alpha-tubulin was observed at all cell cycle stages with the exception of entering mitosis for the former and the end of mitosis for the latter.
Assuntos
Transformação Celular Viral , Centríolos/metabolismo , Microtúbulos/metabolismo , Mitose , Tubulina (Proteína)/metabolismo , Acetilação , Animais , Células 3T3 BALB , Centríolos/ultraestrutura , Camundongos , Microscopia Imunoeletrônica , Microtúbulos/ultraestrutura , Vírus 40 dos SímiosRESUMO
One of the problems of plasma proteomics is a presence of large major components. In this work, we use the thermostable fraction as a way to deplete these major proteins. The thermostable fraction of serum samples from patients with ovarian, uterus, and breast cancers and benign ovarian tumor was analyzed using two-dimensional electrophoresis combined with MALDI-TOF(-TOF)-mass spectrometry. Of them, alpha-1-acid glycoprotein and clusterin are expressly down-regulated in breast cancer, whereas transthyretin is decreased specifically in ovarian cancer. Apolipoprotein A-I forms have decreased spot volumes, while haptoglobin alpha1, in contrast, is elevated in several tumors. These data are partly consistent with previous art studies on cancer proteomics, which involve mass-spectrometry-based serum profiling techniques. Serum thermostable fraction may be recommended as a good tool for medium and small protein proteome investigation, in particular, by 2D-electrophoresis.
Assuntos
Biomarcadores Tumorais/sangue , Eletroforese em Gel Bidimensional/métodos , Proteínas de Neoplasias/sangue , Proteoma/análise , Proteômica/métodos , Adulto , Idoso , Neoplasias da Mama/sangue , Neoplasias da Mama/metabolismo , Feminino , Humanos , Pessoa de Meia-Idade , Neoplasias Ovarianas/sangue , Neoplasias Ovarianas/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Neoplasias Uterinas/sangue , Neoplasias Uterinas/metabolismoRESUMO
The gram-negative bacterium Helicobacter pylori is found in human gastric mucosa. A widely distributed H. pylori infection is associated with chronic gastritis, gastric and duodenal ulcers, and malignant neoplasms. In this study proteome maps of four H. pylori clinical isolates derived from patients of two Russian regions (Moscow/Moscow Region and Novosibirsk) were obtained using 2D-electrophoresis and MALDI-TOF-mass-spectrometry. Variability of some H. pylori proteins and the level of their expression have been evaluated. These four isolates could be easily subdivided into two equal groups characterized by the close proteome profiles and the isolate from Moscow Region and the isolate from Novosibirsk constituted one group. The present study demonstrates the potential of proteome technology, which can be employed together with genome and transcriptome studies for the multiparameter typing of clinical isolates of pathogenic microorganisms.
Assuntos
Infecções por Helicobacter/microbiologia , Helicobacter pylori/química , Mapeamento de Peptídeos , Proteoma , Proteômica , Extratos Celulares/química , Eletroforese em Gel Bidimensional , Genótipo , Helicobacter pylori/genética , Helicobacter pylori/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
New peptidomimetics that have been obtained in the course of our experimental work show distinct insulin-like activity both in vitro and in vivo. The first peptidomimetic (PM 1) is essentially a decapeptide in which sites of A (20-21) and B (19-26) chains of insulin are linked by the peptides bond (Cys-Gly-Glu-Arg-Gly-Phe-Phe-Tyr-Cys-Asn). The second peptidomimetic (PM 2) has similar set of amino acid residues, except that two aromatic amino acids corresponding to the residues of B chain of insulin (B24 and B26) have been replaced with their D optical isomers (Cys-Gly-Glu-Arg-Gly-DPhe-Phe-DTyr-Cys-Asn). The third peptidomimetic (PM 3) has been obtained through acylation of N-terminal of PM 1 by the use of palmitic acid. The peptidomimetic incorporating D aromatic amino acids (PM 2) was demonstrated to exhibit more pronounced hypoglycemic impact, while the acylation of decapeptide tends to prolong the effective time of peptidomimetic influence in vivo.
Assuntos
Insulina/farmacologia , Peptídeos/química , Animais , Glicemia/análise , DNA/biossíntese , Diabetes Mellitus Experimental , Relação Dose-Resposta a Droga , Glucose/farmacocinética , Masculino , Ácido Palmítico/farmacologia , Peptídeos/farmacologia , Estrutura Terciária de Proteína , Ratos , Ratos Wistar , Espectrometria de Fluorescência , Estereoisomerismo , Fatores de TempoRESUMO
The synthesized decapeptide represents functionally important site for binding to the insulin receptor. Amino acid residues at position, 1-8 correlate with B-chain of insulin at position B19-B26, and the residues at position 9-10 correlate with A-chain at position A20-A21. The new peptide was obtained by substitution of two aromatic L-amino acid residues (B24 and B26) for their D-optical isomers. These peptides were tested with cell cultures L929 and PC12 (glucose uptake). Increased concentration of peptides correlated with stimulation of glucose uptake by cells. Studies carried out on animals with streptosotocine-caused diabetes showed that, synthesized peptides were able to decrease glucose level in blood, but decapeptide with D amino acid showed a more pronounced effect compared to the decapeptide with L amino acid.
Assuntos
Hipoglicemiantes/síntese química , Insulina/análogos & derivados , Insulina/síntese química , Substituição de Aminoácidos , Animais , Glicemia/metabolismo , Diabetes Mellitus Experimental/metabolismo , Hipoglicemiantes/química , Hipoglicemiantes/farmacocinética , Insulina/química , Camundongos , Mimetismo Molecular , Ratos , Receptor de Insulina/metabolismo , Estereoisomerismo , Células Tumorais CultivadasRESUMO
Possible employment of cell-specific peptide for the specific adsorption and uptake by cells. It was shown, that apoprotein E 139-158 peptide increases liposomal binding followed by receptor-mediated endocytosis by cells PC12.