protein array
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BACKGROUND: Human colostrum and milk contain components that influence development. Our aim was to use a protein array to determine the cytokine profile of human lacteal secretions and changes that occur during the early postpartum period. METHODS: We collected 17 samples of colostrum during the first 2 days postpartum and a 2nd group of 5 sets of 2 to 3 sequential colostrum or milk samples (at 20- to 30-h intervals). We analyzed the samples with array membranes consisting of 42 or 79 antibodies directed against cytokines. RESULTS: In most samples, we detected the previously described cytokines interleukin-8 (IL-8)/CXCL8, epidermal growth factor (EGF), growth-related oncoprotein (GRO)/CXCL1-3, angiogenin, transforming growth factor beta-2, and monocyte chemotactic protein 1 (MCP-1/CCL2). In addition, we found 32 cytokines that have not been described before in colostrum. Cytokine concentrations differed among mothers, and the spectrum of cytokines changed with time after delivery. A significant decrease occurred in IL-12 and macrophage inflammatory protein-1delta/CCL15 and a significant increase in MCP-1/CCL2. The production of angiogenin, vascular endothelial growth factor, GRO/CXCL1-3, EGF, and IL-8/CXCL8 remained high throughout. The concentrations of 2 selected cytokines measured with the array technique and ELISA showed moderate to strong correlation (r = 0.63 for EGF and r = 0.84 for IL-8/CXCL8). CONCLUSION: Despite the lack of precise quantification, the protein array might be suitable for cytokine screening. It allows simultaneous detection of a broad spectrum of cytokines (including those not described before) in lacteal secretions.
- MeSH
- časové faktory MeSH
- chemokiny analýza MeSH
- čipová analýza proteinů MeSH
- cytokiny analýza MeSH
- dospělí MeSH
- ELISA MeSH
- financování organizované MeSH
- kolostrum chemie MeSH
- lidé MeSH
- mateřské mléko chemie MeSH
- mezibuněčné signální peptidy a proteiny analýza MeSH
- poporodní období MeSH
- proteomika MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- srovnávací studie MeSH
Using the protein array method we determined the serum levels of a number of angiogenic factors. We identified serum levels of angiogenin, PDGF and MCP-1 (CCL2 chemokine) in serum of 32 patients with RCC, and 14 healthy volunteers by means of antibody array analysis. The patients were divided into three groups according to their disease stages (I+II, III, and IV). We found significant differences between the controls and patients with RCC both pre-operatively and post-operatively in angiogenin, PDGF and MCP-1 serum levels. The increase in angiogenin, PDGF and MCP-1 lasted in patients with RCC stages I-III even without metastases eight weeks post-operatively. The patients with stage IV RCC showed disturbed production of PDGF and MCP-1. Protein array analysis is a powerful tool for the identification of large numbers of trace proteins. Multiplex antibody array is able to provide data more precisely reflecting the nature of pathological processes.
- MeSH
- chemokin CCL2 krev MeSH
- čipová analýza proteinů MeSH
- destičkový růstový faktor metabolismus MeSH
- karcinom z renálních buněk chirurgie krev patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- pankreatická ribonukleasa krev MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
Wiley series in probability and statistics
1st ed. xiv, 246 s.
- MeSH
- DNA MeSH
- konformace proteinů MeSH
- Konspekt
- Biochemie. Molekulární biologie. Biofyzika
- NLK Obory
- biochemie
- genetika, lékařská genetika
Goeckerman's therapy (GT) of psoriasis is based on daily application of pharmacy grade coal tar on affected skin with subsequent exposure to UV light. The aim of this study was to evaluate the influence of Goeckerman's therapy of psoriasis on the levels of proangiogenic chemokines ENA-78 (CXCL5, Epithelial Cell Derived Neutrophil Attractant- 78), GRO alpha (CXCL1, Growth-Related Oncogene), IL-8 (CXCL8, Interleukin-8), MCP-1 (CCL2, Monocyte Chemotactic (Chemoattractant) Protein 1) and RANTES (CCL5, Regulated on Activation of Normal T Cell Expressed and Secreted) in peripheral blood of 22 children's patients with psoriasis. 22 otherwise healthy children serve as a control group. The serum levels of chemokines were determined by commercial membrane protein array technique (RayBiotech, USA). Efficacy of Goeckerman's therapy was delineated by PASI score. Disease activity was significantly diminished by Goeckerman's therapy (p<0.001). Serum levels of GRO alpha and MCP-1 in patients before GT were significantly higher than those measured in healthy blood donors (GRO alpha: p=0.0128 and MCP-1: p=0.0003). Serum levels of GRO alpha, MCP-1 and RANTES were significantly diminished by GT (GRO alpha: p=0.002, MCP-1: p=0.048 and RANTES: p=0.0131). Compared to the healthy controls, serum level of MCP-1 remained significantly increased in psoriasis patients after GT (p<0.0001). In conclusion, we found that the GT of psoriasis influenced the serum levels of proinflammatory and proangiogenic chemokines, especially GRO alpha, MCP-1 and RANTES. It could be the cause for decreased proangiogenic activity which is described after GT of psoriasis.
- MeSH
- chemokiny CC krev MeSH
- chemokiny CXC krev MeSH
- čipová analýza proteinů MeSH
- dehet uhelný aplikace a dávkování MeSH
- dítě MeSH
- financování organizované MeSH
- keratolytika aplikace a dávkování MeSH
- lidé MeSH
- mladiství MeSH
- psoriáza krev terapie MeSH
- terapie ultrafialovými paprsky MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
xv, 469 s. : il. ; 26 cm
Sdělení podává informace o novém přístupu k biochemickým analýzám. Dále shrnuje principy, použití, možné výhody či nevýhody nových multiplexních technologií. Většina prezentovaných metod je v současné době buď ve fázi vývoje, nebo je již používána v rámci výzkumných projektů. Pouze některé technologie postupně pronikají do laboratoří klinické biochemie.
The publication reports recent approach to biochemical analyses. There are summarized data about principles, usages, advantages or disadvantages of new multiplex technologies. Although, in the meantime, the most of presented methods are in development or they are used in research projects some technologies already expand to clinical biochemistry laboratories.
Methods in molecular biology ; Vol. 264
[1st ed.] xi, 287 s. : il.
- MeSH
- klinické laboratorní techniky MeSH
- molekulární biologie MeSH
- proteiny MeSH
- Publikační typ
- monografie MeSH
- Konspekt
- Biochemie. Molekulární biologie. Biofyzika
- NLK Obory
- biologie
Cíl sdělení: Byly sledovány analytické a klinické vlastnosti technologie proteinových biočipů pro imunochemická stanovení kardiálních markerů a jejich možné využití v diagnostice infarktu myokardu. Metodika: Stanovení hladiny kardiálních markerů (CKMB, MYO, GPBB, H-FABP, CAIII a cTnI) bylo provedeno systémem Evidence Investigator (Randox). Pro jednotlivé kardiální markery stanovené panelem Cardiac Array byly hodnoceny analytické parametry testu. Kardiální markery byly stanoveny ve vzorcích sér 28 pacientů (21 mužů, 7 žen, věk 47–86 let) s diagnózou akutního infarktu myokardu. Výsledky a závěry: Mezilehlá přesnost pro všechny analyty, kromě myoglobinu, je ve shodě s výrobcem (CV < 10%). H-FABP i GPBB jsou časnými markery akutního infarktu myokardu, přičemž H-FABP má pravděpodobně vyšší diagnostickou senzitivitu než myoglobin. Dá se očekávat, že stanovení kardiálních markerů proteinovými biočipy do budoucna najde své uplatnění v biochemické diagnostice poškození myokardu.
Objective: The analytical and clinical performance of protein biochip technology of immunochemical analysis of cardiac markers and their use in diagnosis of acute myocardial infarction were evaluated. Method: Determination of concentration of cardiac markers (CKMB, MYO, GPBB, H-FABP, CAIII and cTnI) was performed by system Evidence Investigator (Randox). Analytical parameters of Cardiac array were tested. Markers of myocardial injury were determined in group of the 28 patients (21 men, 7 women, age 47–86 years) with acute myocardial infarction diagnosis. Results and conclusions: Reproducibility of all tests agree with data of producer (CV < 10%), except myoglobin assay. New substances H-FABP and GPBB are promising early markers of acute myocardial infarction and diagnostic sensitivity of H-FABP would be better than myoglobin test. In future, determination of new cardiac markers by protein biochips technology probably will be used in cardiologic diagnosis.
- MeSH
- biologické markery krev MeSH
- čipová analýza proteinů přístrojové vybavení využití MeSH
- financování organizované MeSH
- glykogenfosforylasa krev MeSH
- infarkt myokardu krev MeSH
- karboanhydrasa III krev MeSH
- lidé MeSH
- nekróza krev MeSH
- proteiny vázající mastné kyseliny krev MeSH
- sérum MeSH
- troponin krev MeSH
- Check Tag
- lidé MeSH
BACKGROUND: The association between increases in cardiac troponin and adverse cardiac outcomes is well established. There is a growing interest in exploring routine cardiac troponin monitoring as a potential early indicator of adverse heart health trends. Prognostic use of cardiac troponin measurements requires an assay with very high sensitivity and outstanding analytical performance. We report development and preliminary validation of an investigational assay meeting these requirements and demonstrate its applicability to cohorts of healthy individuals and patients with heart failure. METHODS: On the basis of single molecule array technology, we developed a 45-min immunoassay for cardiac troponin I (cTnI) for use on a novel, fully automated digital analyzer. We characterized its analytical performance and measured cTnI in healthy individuals and heart failure patients in a preliminary study of assay analytical efficacy. RESULTS: The assay exhibited a limit of detection of 0.01 ng/L, a limit of quantification of 0.08 ng/L, and a total CV of 10% at 2.0 ng/L. cTnI concentrations were well above the assay limit of detection for all samples tested, including samples from healthy individuals. cTnI was significantly higher in heart failure patients, and exhibited increasing median and interquartile concentrations with increasing New York Heart Association classification of heart failure severity. CONCLUSIONS: The robust 2-log increase in sensitivity relative to contemporary high-sensitivity cardiac troponin immunoassays, combined with full automation, make this assay suitable for exploring cTnI concentrations in cohorts of healthy individuals and for the potential prognostic application of serial cardiac troponin measurements in both apparently healthy and diseased individuals.
- MeSH
- čipová analýza proteinů ekonomika metody MeSH
- dospělí MeSH
- imunoanalýza ekonomika metody MeSH
- lidé středního věku MeSH
- lidé MeSH
- limita detekce MeSH
- mladý dospělý MeSH
- srdeční selhání krev diagnóza MeSH
- troponin I analýza krev MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH