- Publikační typ
- abstrakt z konference MeSH
Jako autoimunitní označujeme ty choroby, které jsou způsobeny reakcí imunitního systému proti zdravým, tělu vlastním strukturám. Diagnostika takovýchto stavů vyžaduje souhru klinické části s částí laboratorní. Tento text se zaměřuje na diagnostiku vybraných systémových a orgánově specifických autoimunitních chorob. Neklade si ale za cíl podrobně popsat jednotlivé choroby, spíše shrnuje nejvýraznější vodítka vedoucí k jejich správné diagnóze. Rovněž nastiňuje používané laboratorní metody a shrnuje jejich výhody a nevýhody.
Autoimmune diseases are caused by reaction of immune system against own body´s healthy structures. Diagnostics of these states require collaboration between clinical and laboratory diagnostic part. This text is focusing on diagnostics selected systemic and organ specific autoimmune diseases. It does not aim for detail description of selected diseases rather summarizing the main clues leading to right diagnostics. Also suggesting laboratory methods used in diagnostics and summarizing their benefits and drawbacks.
- MeSH
- autoimunitní nemoci * diagnóza MeSH
- imunoanalýza metody MeSH
- klinické laboratorní techniky metody MeSH
- lidé MeSH
- protilátky analýza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are two enzymes sensitive to various chemical compounds having ability to bind to crucial parts of these enzymes. Boldine is a natural alkaloid and it was mentioned in some older works that it can inhibit some kinds of AChE. We reinvestigated this effect on AChE and also on BChE using acetyl (butyryl) thiocholine and Ellman's reagents as standard substances for spectrophotometric assay. We found out IC50 of AChE equal to 372 μmol/l and a similar level to BChE, 321 μmol/l. We conclude our experiment by a finding that boldine is cholinesterase inhibitor; however we report significantly weaker inhibition than that suggested in literature. Likewise, we tried to investigate the mechanism of inhibition and completed it with in silico study. Potential toxic effect on cholinesterases in real conditions is also discussed.
- MeSH
- acetylcholinesterasa metabolismus MeSH
- aporfiny chemie farmakologie MeSH
- butyrylcholinesterasa chemie metabolismus MeSH
- cholinesterasové inhibitory chemie farmakologie MeSH
- HEK293 buňky MeSH
- lidé MeSH
- molekulární modely MeSH
- sekundární metabolismus * MeSH
- substrátová specifita účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Biperiden is a drug used in Parkinson disease treatment and it serves also as an antiseizures compound in organophosphates poisoning. It acts as antagonist of muscarinic receptor activated by acetylcholine while the enzyme acetylcholinesterase (AChE) cleaves acetylcholine in synaptic junction into choline and acetic acid. This enzyme is inhibited by various compounds; however there has not been proposed evidence about interaction with biperiden molecule. We investigated this interaction using standard Ellman's assay and experimental findings were critically completed with an in silico prediction by SwissDock docking software. Uncompetitive mechanism of action was revealed from Dixon plot and inhibition constant (Ki ) was calculated to be 1.11 mmol/l. The lowest predicted binding energy was -7.84 kcal/mol corresponding to H-bond between biperiden molecule and Tyr 341 residuum in protein structure of AChE. This interaction seems to be further stabilized by π-π interaction with Tyr 72, Trp 286, and Tyr 341. In conclusion, biperiden appears as a very weak inhibitor but it can serve as a lead structure in a pharmacological research.
- MeSH
- acetylcholinesterasa metabolismus MeSH
- biperiden chemie farmakologie terapeutické užití MeSH
- cholinesterasové inhibitory chemie farmakologie terapeutické užití MeSH
- enzymatické testy MeSH
- lidé MeSH
- molekulární modely MeSH
- Parkinsonova nemoc farmakoterapie enzymologie MeSH
- substrátová specifita účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Magnetic particles (MPs) have been widely used in biological applications in recent years as a carrier for various molecules. Their big advantage is in repeated use of immobilized molecules including enzymes. Acetylcholinesterase (AChE) is an enzyme playing crucial role in neurotransmission and the enzyme is targeted by various molecules like Alzheimer's drugs, pesticides and warfare agents. In this work, an electrochemical biosensor having AChE immobilized onto MPs and stabilized through glutaraldehyde (GA) molecule was proposed for assay of the neurotoxic compounds. The prepared nanoparticles were modified by pure AChE and they were used for the measurement anti-Alzheimer's drug galantamine and carbamate pesticide carbofuran with limit of detection 1.5 µM and 20 nM, respectively. All measurements were carried out using screen-printed sensor with carbon working, silver reference, and carbon auxiliary electrode. Standard Ellman's assay was used for validation measurement of both inhibitors. Part of this work was the elimination of reversible inhibitors represented by galantamine from the active site of AChE. For this purpose, we used a lower pH to get the original activity of AChE after inhibition by galantamine. We also observed decarbamylation of the AChE-carbofuran adduct. Influence of organic solvents to AChE as well as repeatability of measurement with MPs with AChE was also established.
A method is described for the determination of fatty acids in dried sweat spot and plasma samples using gas chromatography with flame ionization detection. Plasma and dried sweat spot samples were obtained from a group of blood donors. The sweat was collected from each volunteer during exercise. Sweat was spotted onto collection paper containing butylated hydroxytoluene. Fatty acids were derivatized with acetyl chloride in methanol to form methyl esters of fatty acids. The fatty acids in dried sweat spot samples treated with butylated hydroxytoluene and stored at -20°C were stable for 3 months. Our results indicate that sweat contains, among fatty acids with short chain, also fatty acids with long chain and unsaturated fatty acids. Linear relationships between percentage content of selected fatty acids in dried sweat spot and plasma were observed.
- MeSH
- chromatografie plynová * MeSH
- estery analýza MeSH
- lidé MeSH
- mastné kyseliny analýza MeSH
- plamínková ionizace * MeSH
- pot chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- Alzheimerova nemoc diagnóza farmakoterapie MeSH
- chemické bojové látky MeSH
- cholinesterasové inhibitory * farmakologie terapeutické užití MeSH
- cholinesterasy * farmakologie terapeutické užití MeSH
- diagnostické techniky a postupy MeSH
- lidé MeSH
- myasthenia gravis diagnóza farmakoterapie MeSH
- otrava * diagnóza MeSH
- pesticidy MeSH
- Check Tag
- lidé MeSH
