Esenciální aminokyselina tryptofan slouží jako substrát pro tvorbu řady bioaktivních sloučenin. V anabolických procesech je využívána jen malá část tryptofanu, zatímco velká většina je metabolizována kynureninovou cestou. Degradace tryptofanu na kynurenin je v současné době chápána jako klíčový regulátor vrozené a adaptivní imunity. Hlavním kontrolním enzymem této dráhy je indolamin 2,3-dioxygenáza (IDO). Jedná se o enzym, který katalyzuje počáteční krok při jeho přeměně na kynurenin. Jednou z fyziologických rolí tohoto enzymu je regulovat zánětlivý proces a mírnit jeho případné nežádoucí účinky. Deplece tryptofanu a přítomnost kynureninů vede k anergii efektorových T lymfocytů, aktivaci T regulačních buněk a může stimulovat dendritické buňky k diferenciaci na imunosupresivní fenotyp. Zvýšená degradace tryptofanu je pozorována u nemocí a poruch provázených buněčnou imunitní aktivací, např. infekčních a autoimunitních onemocnění, alergických stavů a nádorových onemocnění. Stanovení hladiny tryptofanu nebo jeho metabolitů by mohlo být přínosné z hlediska monitorování některých imunopatologických stavů. Problémem však zůstává technická dostupnost laboratorního stanovení tohoto parametru. Aktivitu IDO lze stanovit i nepřímo poměrem kynureninu a tryptofanu, což je spolehlivý index degradace tryptofanu.
The essential amino acid tryptophan is a substrate for the generation of several bioactive compounds with important physiological roles. Only a small fraction of ingested tryptophan is used in anabolic processes, whereas the large majority is metabolized along the kynurenine pathway of tryptophan degradation. This pathway generates a range of metabolites, collectively known as kynurenines, involved in inflammation and immune response. The kynurenine pathway is highly regulated in the immune system, where it promotes immunosuppression in response to inflammation or infection. Kynurenine reduces the activity of natural killer cells, dendritic cells, or proliferating T cells. Indoleamine 2,3-dioxygenase is a rate-limiting enzyme for tryptophan metabolism. Increased tryptophan degradation is observed in diseases and disorders accompanied by cellular immune activation, such as infectious and autoimmune diseases, allergic conditions and cancer. Determination of the level of tryptophan or its metabolites could be beneficial in terms of monitoring certain immunopathological conditions. Actually, indoleamine 2,3-dioxygenase activity could be indirectly measured by the kynurenin to tryptophan ratio, a reliable index of tryptophan breakdown
- MeSH
- Immunity, Active physiology immunology MeSH
- Hypersensitivity enzymology immunology MeSH
- Autoimmune Diseases epidemiology immunology metabolism MeSH
- Immune Tolerance immunology MeSH
- Indoleamine-Pyrrole 2,3,-Dioxygenase physiology immunology MeSH
- Kynurenine metabolism MeSH
- Humans MeSH
- Neoplasms metabolism MeSH
- Gastrointestinal Microbiome immunology MeSH
- Tryptophan * analysis immunology metabolism MeSH
- Chromatography, High Pressure Liquid methods MeSH
- Check Tag
- Humans MeSH
- Publication type
- Review MeSH
Závěrečná zpráva o řešení grantu Interní grantové agentury MZ ČR
1 svazek : ilustrace, tabulky ; 30 cm
Parkinsonský syndrom je charakterizován bradykinezí, klidovým třesem, rigiditou a posturálními poruchami. Jeho nejčastější příčinou (asi 80% případů) je idiopatická Parkinsonova nemoc. Ale také celá řada dalších neurodegenerativních onemocnění, jako např. demence s Lewyho tělísky, progresivní supranukleární paralýza, multisystémová atrofie, kortikobazální degenerace a jiné., se mohou manifestovat parkinsonským syndromem. Diferenciální diagnostika mezi jednotlivými onemocněními je značně limitována. Cílem projektu bude u pacientů s příznaky parkinsonského syndromu a u věkově identické kontrolní skupiny provést podrobné vyšetření séra a mozkomíšního moku se stanovením hladin markerů neurodegenerace, zánětu, onkomarkrů, ukazetelů tkáňové destrukce a dalších. Hladiny těchto markerů budou nadále po odběru srovnávány s průběhem onemocnění, jeho případnou progresí či vývoj v jiné onemocnění. Získané informace by poté mohly vést k určení možného prediktivního faktoru určujícího diagnózu onemocnění.; Parkinson syndrome (PS) is characterised by bradykinesis, rest tremor, rigidity and postural instability. The most frequent cause (about 80%) is idiopatic Parkinson's disease. But also a number of other neurodegenerative diseases , e.g. Lewy body disease, progressive supranuclear palsy, multisystem atrophy, corticobasal degeneration etc., can manifest itself with PS. Differential diagnosis between individual diseases is highly limited. Project goal will be in case of patients with PS and age identical control group to do detailed examination of serum and cerebrospinal fluid with determination of levels of neurodegenerative markers, inflamattory markers, oncomarkers, histic destruction pointers and other. The levels of the markers we took will be from now on compared with the disease process, its possible progression or development in different diseases. Gathered information could be instrumental towards the asessment of possible predictive factor defining disease prediction.
- MeSH
- Diagnosis, Differential MeSH
- Phenotype MeSH
- Cerebrospinal Fluid cytology MeSH
- Neurodegenerative Diseases diagnosis classification MeSH
- Parkinson Disease diagnosis MeSH
- Tandem Mass Spectrometry methods utilization MeSH
- Tryptophan analogs & derivatives analysis MeSH
- Conspectus
- Patologie. Klinická medicína
- NML Fields
- neurologie
- NML Publication type
- závěrečné zprávy o řešení grantu IGA MZ ČR
AIM: A new HPLC method for the determination of neopterin, kynurenine and tryptophan using a second-generation monolith stationary phase and high-throughput sample preparation procedure based on microplates was developed and fully validated. MATERIALS & METHODS: As the stationary phase a monolithic C18 Chromolith high-resolution column with dimensions of 4.6 × 100 mm connected to a monolithic 4.6 × 10-mm security guard was used. Separation was achieved using 15 mM phosphate buffer (KH2PO4 +K2HPO4·3H2O at pH 3) and acetonitrile in gradient mode. RESULTS: Target analytes were determined in 5.5 minutes in amniotic fluid, effusions and wound exudates with a limit of quantification (LOQ) of 1.25 nM for neopterin, 2.5 µM for tryptophan and 0.25 µM for kynurenine. DISCUSSION: The method was applied to real clinical sample measurements, and it will be used to monitor neopterin, kynurenine and tryptophan levels in biological fluids to assess the patient response to therapy and clinical status.
- MeSH
- Exudates and Transudates metabolism MeSH
- Wound Healing * MeSH
- Kynurenine analysis MeSH
- Humans MeSH
- Pleural Effusion, Malignant metabolism MeSH
- Neopterin analysis MeSH
- Amniotic Fluid metabolism MeSH
- Spectrophotometry, Ultraviolet MeSH
- Tryptophan analysis MeSH
- Chromatography, High Pressure Liquid methods MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Human milk is a living fluid that changes with time, composition and volume. Circadian rhythms regulate a variety of biological processes in living organisms; and perhaps the most evident function is the sleep-wake cycle. The aim of the present study was to evaluate the circadian rhythm of breast milk amino acids and their evolution throughout the breastfeeding period. Human breast milk samples from 77 donors were collected every 3 hours over a 24-h period. The rhythmicity of the amino acids was determined by cosinor analysis. Colostrum samples showed no circadian rhythm in most amino acids except tryptophan. However, daily variations were observed in tryptophan and methionine at transitional phase, according to the newborn’s pattern of intake every 3 hours regardless of whether it is day or night. During the last stage (mature milk), when breast milk has fully stabilized, most amino acids showed a circadian rhythm. In conclusion, breast milk should be given to the baby at the same time of day it is expressed. Thus, the baby would be adjusting its circadian pattern in harmony with his environment (day/night), which is crucial for the proper functioning and synchronization of all systems in the human body.
- MeSH
- Amino Acids * analysis metabolism MeSH
- Chronobiology Phenomena MeSH
- Circadian Rhythm * physiology MeSH
- Confidence Intervals MeSH
- Cohort Studies MeSH
- Breast Feeding MeSH
- Colostrum chemistry metabolism MeSH
- Lactation metabolism MeSH
- Humans MeSH
- Milk, Human * chemistry metabolism MeSH
- Methionine analysis metabolism MeSH
- Reproducibility of Results MeSH
- Statistics as Topic MeSH
- Tandem Mass Spectrometry MeSH
- Tryptophan analysis metabolism MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
A complex of osmium tetroxide with 2,2'-bipyridine (Os,bipy) has been applied as a chemical probe of DNA structure as well as an electroactive DNA label. The Os,bipy has been known to form covalent adducts with pyrimidine DNA bases. Besides the pyrimidines, electrochemically active covalent adducts with Os,bipy are formed also by tryptophan (W) residues in peptides and proteins. In this paper we show that Os,bipy-treated proteins possessing W residues (such as avidin, streptavidin, or lysozyme) yield at the pyrolytic graphite electrode (PGE) a specific signal (peak alphaW) the potential of which differs from the potentials of signals produced by free Os,bipy or by Os,bipy-modified DNA. No such signal is observed with proteins lacking W (such as ribonuclease A or alpha-synuclein). Subpicomole amounts of W-containing proteins modified with Os,bipy can easily be detected using adsorptive transfer stripping voltammetry with the PGE. Binding of biotin to avidin interferes with Os,bipy modification of the protein, in agreement with the location of W residues within the biotin-binding site of avidin. These Ws are accessible for modification in the absence of biotin but hidden (protected from modification) in the avidin-biotin complex. The Os,bipy-modified avidin is unable to bind biotin, and its quarternary structure is disrupted. Analogous effects were observed with another biotin-binding protein, streptavidin. Our results demonstrate that modification of proteins with Os,bipy under conditions close to physiological, followed by a simple electrochemical analysis, can be applied in the microanalysis of protein structure and interactions.
- MeSH
- 2,2'-Dipyridyl chemistry MeSH
- Avidin chemistry MeSH
- Biotin chemistry MeSH
- Electrochemistry MeSH
- Electrodes MeSH
- Electrons MeSH
- Financing, Organized MeSH
- Molecular Structure MeSH
- Osmium Tetroxide analysis chemistry MeSH
- Proteins analysis chemistry MeSH
- Cross-Linking Reagents chemistry MeSH
- Tryptophan analysis chemistry MeSH
- Carbon chemistry MeSH
Liečba zvierat s ischemicko-reperfúznym poškodením mozgu použitím antioxidantov, vychytávačov voľných radikálov, by mohla znížiť mieru závažnosti reperfúzneho poškodenia. V našich experimentoch sme overovali protektívny účinok extraktu z Ginkgo biloba (EGb) na I/R indukovaný oxidačný stres a oxidáciu proteínov. Pozorovali sme signifikantný nárast intenzity fluorescencie tryptofánu po 24 hod. reperfúzii (24 hod. REP) u zvierat s podávaným EGb v porovnaní s 24h REP skupinou bez EGb. Signifikantný pokles intenzity fluorescencie dityrozínu u kontrolných, ischemických a 24 hod. REP skupín s EGb boli pozorované v porovnaní so skupinami bez EGb. Naše výsledky naznačujú, že extrakt z Ginkgo biloba ako predpokladný antioxidant môže zohrávať dôležitú úlohu pri znižovaní ischémiou/repefúziou indukovaného oxidačného stresu a proteínovej oxidácie.
Treatment of the animals with brain ischemic-reperfusion injury using antioxidants, scavengers of free radicals may reduce the severity of reperfusion damage. The present study was designed to check the protective effect of extract of Ginkgo biloba (EGb) on I/R injury induced oxidative stress and protein oxidation. We have observed significant increase of tryptophan fluorescence intensity of 24 h reperfusion (REP) groups with EGb administration in comparison to 24 h REP groups without EGb. Significant decrease of bityrosine fluorescence intensity in control groups, ischemia, 24 h REP with EGb were observed in comparison to groups without EGb. Our results indicate that extract of Ginkgo biloba as a presumed antioxidant could play an important role in reducing the ischemia/reperfusion-induced oxidative stress and protein oxidation.
- MeSH
- Child MeSH
- Research Support as Topic MeSH
- Skin Diseases metabolism MeSH
- Humans MeSH
- Adolescent MeSH
- Photosensitivity Disorders metabolism MeSH
- Tryptophan analogs & derivatives analysis metabolism MeSH
- Metabolism, Inborn Errors metabolism MeSH
- Chromatography, High Pressure Liquid methods MeSH
- Check Tag
- Child MeSH
- Humans MeSH
- Adolescent MeSH
