Determination of biomarkers in body fluids provides information about the overall state of the organism 1. Among commonly determined markers in clinical laboratories belong AAs. The AAs (Fig. 1), basic structural units of peptides, as well as their derivatives occupy many metabolic and biochemical roles in human body 2,3. The AAs belong to zwitterions containing acidic and basic functional groups (carboxyl groups and amino, respectively). Zwitterions have neutral character at a certain pH known as the isoelectric point due the dissociation of acidic and basic groups. Therefore, the charge of AAs depend on the number of functional groups and pH environment 4. Determination of AAs levels in the body fluids such as blood and/or plasma 5,6, cerebrospinal fluid 6,7, urine 6,8, saliva 6,7 or amniotic fluid 9 represents the significant clinical indicator not only for control of the nutritional state of the organism, but for a number of metabolic disorders 10. Examples can be inherited metabolic disorders such as phenylketonuria (PKU) (an enzyme deficiency in phenylalanine hydroxylase (PAH) (Fig. 2)) 11 or maple syrup urine disease (an enzyme deficiency in branched-chain α-ketoacid dehydrogenase (BCKD) (Fig. 3)) 12. There are many methods for AAs determination employing various techniques such as high pressure liquid chromatography (HPLC) 13 or gas chromatography (GC) 14. During the 80's of the 20th century CE became a routine laboratory technique 15. Recently, this method has ranked among very useful tools for separation and determination of AAs 2,9,16,17, biogenic amines 18 or peptides 19 and is often used as an alternative to HPLC or GC due to its electrophoretic separation mechanism 16,20. In addition, a main advantage is very low sample and solvent consumption, speed of analysis and excellent separation efficiency 2,16,21-23. Besides, due to the high resolving power is CE a promising method for the analysis of biological fluids (complex mixtures of many metabolites) 2,22. The essential part of the CE instrumentation is a detector. In photometric and fluorescence detection is problem with a limited presence of chromophores and fluorofores. Because of that, the derivatization procedures prior CE are usually used, which is the one of the limiting factors 24. Therefore, there are attempts to analyze AAs without derivatization employing various strategies such as indirect detection 25, electrochemical detection (conductometric detection 6,8, amperometric detection 26) or CE in combination with mass spectrometry (MS) 5,11.
- MeSH
- aminokyseliny * krev metabolismus moč MeSH
- biologické markery MeSH
- elektroforéza kapilární * MeSH
- lidé MeSH
- tělesné tekutiny * chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
The microRNA (miRNA) belongs to short noncoding RNA molecules (generally 20-25 nt in length) which are able of specific gene regulation expression 1. These regulatory molecules have significant influence on different biological processes like cell proliferation, development, differentiation, apoptosis and metabolism 2. The aberrant expression of various miRNAs is involved in the cell pathological status formation. The abnormal miRNA expression levels were found at number diseases like inflammatory and autoimmune diseases, diabetes, cardiovascular and neurodegenerative diseases and cancer 3-6. Specific miRNAs expression profiles were identified at various tumors so miRNA are interesting diagnostic biomarkers. Moreover their detection might be helpful to find more about cancer staging, prognosis and/or response to treatment 7. Different studies have shown that miR-150 is upregulated in lung cancer tissue and lupus nephritis, downregulated at patients with heart failure, acute myeloblastic leukemia, colorectal cancer and systemic sclerosis 8-14. Because of miRNAs importance the simple and fast detection method is needed. Therefore, the sensitive electrochemical analysis was combined with specific magnetic particles-based isolation.
MicroRNAs are small non-coding RNA molecules that are involved in post-transcriptional regulation of gene expression. They take up about 2 % of the genome and affects up to one third of the protein-coding genes. Their biogenesis starts in the cell nucleus and passes into the cytoplasm where the functional forms of miRNAs participate in the process of RNA interference. Nowadays, the biggest interest focuses on the role of miRNAs during carcinogenesis. Mutations in the genes for various miRNAs types is to promote proliferation and inhibition of apoptosis. Therefore, the miRNA counts as a good diagnostic marker or even as a treatment method.
- MeSH
- biogeneze organel MeSH
- lidé MeSH
- mikro RNA * genetika MeSH
- nádory genetika MeSH
- RNA interference MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
Dot-blot is one of biological methods that are normally used in research laboratories and especially in the diagnostics. It is the most commonly used method for identification and immunodetection of particular proteins which may be markers of various diseases. The main aim of the experiment was to develop a simple, inexpensive and rapid method for specific detection of nucleic acids, especially DNA, and then this procedure apply to the detection of DNA modified by platinum cytostatic drugs. Despite platinum cytostatic drugs‘ common use in chemotherapy of various cancer types, their biochemical effect is still not completely clear. The generally accepted opinion is that the drug binds to cellular DNA. It was observed that cisplatin is bound to the DNA the most compared to oxaliplatin and carboplatin.
- Klíčová slova
- oxaliplatina,
- MeSH
- antitumorózní látky * MeSH
- cisplatina MeSH
- DNA * imunologie účinky léků MeSH
- imunoblotting * metody statistika a číselné údaje MeSH
- karboplatina MeSH
- organoplatinové sloučeniny MeSH
- polymerázová řetězová reakce MeSH
- protilátky MeSH
- Publikační typ
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- MeSH
- elektroforéza kapilární * MeSH
- lidé MeSH
- mikro RNA * MeSH
- nádorové biomarkery genetika MeSH
- nádory prostaty * genetika MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- práce podpořená grantem MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
