RESUMEN
Under controlled conditions of 17-day isolation (Sirius-17 experiment), the protein composition of urine was studied in 6 healthy test volunteers-3 women and 3 men. Collection of samples in the form of a second freely separated morning urine fraction was carried out in the background (seven days before the experiment), as well as 1 day after the end of exposure. Chromatographic-mass-spectrometric semi-quantitative analysis of the protein composition of samples was performed on a system consisting of an Agilent 1100 chromatograph and an LTQ-FT Ultra hybrid mass spectrometer using bioinformatics resources UniProtKB, GeneOntology. An asymptomatic change in the immune defense system of kidney tissue after isolation in a closed hermetic object is associated with a change in the content of 7 proteins that provide functional activity of the TLR tubules of the kidneys - FcRIII, MUC1, Galectin-3, Ficolin-2, APOA1, FLNA, FCGR3A and Clusterin. These proteins are found to be useful biomarkers in the study of physiology and kidney diseases. They can be attributed to candidates for protein markers of the initial stages of impaired recognition by the epithelium of renal tubules of bacteria with known pathogenic potential.
Asunto(s)
Espectrometría de Masas , Proteoma , Urinálisis , Biomarcadores , Cromatografía , Humanos , Masculino , Receptores de IgGRESUMEN
Clinical observation and examination of 12 patients with chronic pyelonephritis (CPN) were performed. The first group (GI) included patients with exacerbation of the disease. In the comparison group (GII)- the same patients after 1.5-3 months after completion of treatment, without clinical manifestations of exacerbation of CPN. Laboratory signs of acute renal damage were not revealed in all examined patients. Additionally, urine was collected in the afternoon after Breakfast, in the form of a freely separated 2nd fraction and its sample preparation, consisting of the stages: recovery, alkylation, protein deposition and proteolysis using trypsin. The resulting polypeptide mixture was separated by liquid chromatography in three repetitions and analyzed on a system consisting of Agilent 1100 chromatograph and ltq-FT ultra hybrid mass spectrometer. A list of proteins was obtained, indicating the number of peptides by which they were identified, and the parameters of its reliability. Most of the information about the obtained proteins was obtained from UniProt databases. Identified and analyzed 10 proteins that differ significantly in occurrence in the clinical group of patients in the period of exacerbation of PN. The appearance of these proteins in urine in 1patients with exacerbation of chronic PH allows us to consider them as potential biomarkers directly associated with inflammation and damage to the epithelial lining of the renal tubules.
Asunto(s)
Proteoma , Pielonefritis/orina , Cromatografía Liquida , Humanos , Espectrometría de Masas , Proteinuria , Proteómica , Reproducibilidad de los Resultados , UrinálisisRESUMEN
We investigated changes in the urine protein composition of healthy volunteers in controlled conditions during 105-day isolation (Mars-500 program) at different levels of salt consumption. We used newest proteomic techniques based on chromatography-mass spectrometry and various methods of bioinformatics including opoSOM. The period of observation can be divided into three intervals with different dynamics of protein excretion: early (week 1-6), intermediate (week 7-11) and late interval (week 12-15). We identified about 10 different groups of co-detected proteins, which are directly affected by periods with different-levels of salt consumption. We also determined the biological functions of these proteins, tissue specificity and signaling pathways that involve them.
Asunto(s)
Proteínas/análisis , Cloruro de Sodio Dietético/administración & dosificación , Orina/química , Adulto , Ambiente Controlado , Cromatografía de Gases y Espectrometría de Masas , Humanos , ProteómicaRESUMEN
Mass spectromy-based proteomics was employed to analyze urine from 8 normal volunteers for a 21-day bedrest study (BR). The analysis included trypsinolysis in solution prior to liquid chromatography and tandem mass spectrometry (LC-MS/MS), and spectra processing using the bioinformatics tools. Relying on 221 IPI-indices with Score from 24 to 1700, 169 different proteins were identified. Molecular functions, biological processes and cell components as the loci of certain protein functioning were determined with the help of UniProt-GOA. Associative interactions networks were constructed using BiNGO. There were 14 proteins identified that are functional in the cardiovascular system mostly. They were annotated and -dynamics of their occurrence throughout the experiment was considered. Grounding on the biological functions of these proteins, an assumption of eligible activation of different biological processes during BR was made.