- Publikační typ
- abstrakt z konference MeSH
OBJECTIVES: With over 35 million cases worldwide, Alzheimer's disease (AD) represents the main cause of dementia. The differentiation of AD from other types of dementia is challenging and its early diagnosis is complicated. The established biomarkers are not only based on the invasive collection of cerebrospinal fluid, but also lack sufficient sensitivity and specificity. Therefore, much current effort is aimed at the identification of new biomarkers of AD in peripheral blood. DESIGN AND METHODS: We focused on blood-based analyses using chiroptical spectroscopy (Raman optical activity, electronic circular dichroism) supplemented with conventional vibrational spectroscopy (infrared, Raman) and metabolomics (high-performance liquid chromatography with a high-resolution mass detection). RESULTS: This unique approach enabled us to identify the spectral pattern of AD and variations in metabolite levels. Subsequent linear discriminant analysis of the spectral data resulted in differentiation between the AD patients and control subjects. CONCLUSIONS: It may be stated that this less invasive approach has strong potential for the identification of disease-related changes within essential plasmatic biomolecules and metabolites.
- MeSH
- Alzheimerova nemoc krev diagnóza MeSH
- biologické markery krev MeSH
- cirkulární dichroismus MeSH
- diskriminační analýza MeSH
- krevní proteiny analýza MeSH
- lidé středního věku MeSH
- lidé MeSH
- metabolomika MeSH
- Ramanova spektroskopie MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- Publikační typ
- abstrakt z konference MeSH
The aim of the presented work was to develop a highly sensitive, accurate and rapid analytical method for the determination of concentration levels of tryptophan and its metabolites of kynurenine catabolic pathway, as well as neurotransmitters and their metabolites in complex biological matrices (brain tissue and blood plasma). The developed analytical method consists of analytes separation from the biological matrices by protein precipitation (blood plasma) or solvent extraction (brain tissue), derivatization of the analytes and their detection by high-performance liquid chromatography combined with mass spectrometry. Individual steps of the whole process were optimized and the method was validated in the terms of selectivity, linearity (R2≥0.980), precision (RSD ≤ 13.3%), recovery (≥82.0%), limit of detection (1.8 ng/mL of blood plasma, 2.2 pg/mg of brain tissue) and limit of quantification (2.5 ng/mL of blood plasma, 2.8 pg/mg of brain tissue). The method was subsequently verified by an animal study, where the concentration levels of the analytes in biological matrices (blood plasma and brain tissue) of T. gondii - infected rats and control animals were compared. All the data obtained from the animal study were statistically evaluated. Increased concentration levels of kynurenine catabolic pathway metabolites (e.g. kynurenine, 3-hydroxykynurenine, quinolinic acid) were observed in the case of T. gondii - infected rats in contrast to the control group. The opposite effect was determined in the case of serotonin and its metabolite 5-hydroxyindoleacetic acid, where higher concentration levels were found in blood plasma of healthy subjects. Finally, Principal Component Analysis (PCA) was utilized for a score plot formation. PCA score plots have demonstrated the similarities of individuals within each group and the differences among the groups.
- MeSH
- krevní plazma metabolismus MeSH
- krysa rodu rattus MeSH
- kynurenin analogy a deriváty krev metabolismus MeSH
- kyselina chinolinová krev metabolismus MeSH
- kyselina hydroxyindoloctová krev metabolismus MeSH
- mozek metabolismus MeSH
- neurotransmiterové látky metabolismus MeSH
- potkani Wistar MeSH
- serotonin metabolismus MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- toxoplazmóza krev metabolismus MeSH
- tryptofan metabolismus MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
To enable the early diagnosis of pancreatic cancer, the search for and definition of reliable biomarkers remain a subject of great interest, with the specificity and sensitivity of the currently used biomarkers being below the required values. We tested a novel diagnostic approach for pancreatic cancer based on the specific molecular signature of blood plasma components. To acquire more detailed structural information, structure-sensitive chiroptical methods (electronic circular dichroism and Raman optical activity) were supplemented by conventional Raman and infrared spectroscopies. The obtained spectra were subsequently processed by linear discriminant analysis yielding high values of specificity and sensitivity. In addition, to monitor not only large biomolecules as potential biomarkers but also those of low molecular weight, we conducted an analysis of blood plasma samples by using metabolomics. The achieved results suggest a panel of promising biomarkers for a reliable detection of pancreatic cancer.
- MeSH
- cirkulární dichroismus * metody MeSH
- diskriminační analýza MeSH
- karnitin analogy a deriváty krev MeSH
- lidé středního věku MeSH
- lidé MeSH
- lysofosfatidylcholiny krev MeSH
- metabolomika * metody MeSH
- nádorové biomarkery krev MeSH
- nádory slinivky břišní * krev MeSH
- pilotní projekty MeSH
- Ramanova spektroskopie * metody MeSH
- senioři MeSH
- spektroskopie infračervená s Fourierovou transformací * metody MeSH
- studie případů a kontrol MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
Diagnostic methods used in the contemporary medical practice consist of a combination of distressing invasive (bronchial biopsy, bronchoalveolar lavage) and semiinvasive (induced sputum technique) methods. Monitoring of specific molecules produced during pathological processes in biological matrices is a relatively new technique which represents an alternative, entirely non-invasive and comfortable method. The principle is based on the quantification of specific substances – "biomarkers", which are considered to be objectively measurable indicators of a physiological/pathological condition of the organism. In contrast with non-specific matrices such as blood plasma (reflecting the state of the whole organism), the exhaled breath condensate is a specific matrix. Concentration levels of biomarkers in the latter matrix point at pathological processes only in the airways and lungs. Typical molecules to be monitored include hydrogen peroxide, leukotrienes, prostaglandins, lipoxins and prostanoids.
- Klíčová slova
- kondenzát vydechovaného vzduchu,
- MeSH
- biologické markery analýza MeSH
- chronická obstrukční plicní nemoc diagnóza MeSH
- diagnostické techniky dýchacího ústrojí MeSH
- ELISA MeSH
- hmotnostní spektrometrie MeSH
- kyseliny prostanové analýza MeSH
- leukotrieny analýza MeSH
- lidé MeSH
- lipoxiny analýza MeSH
- metody pro podporu rozhodování MeSH
- prognóza MeSH
- prostaglandiny analýza MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- Publikační typ
- abstrakt z konference MeSH
Perinatal immune challenge leads to neurodevelopmental dysfunction, permanent immune dysregulation and abnormal behaviour, which have been shown to have translational validity to findings in human neuropsychiatric disorders (e.g. schizophrenia, mood and anxiety disorders, autism, Parkinson's disease and Alzheimer's disease). The aim of this animal study was to elucidate the influence of early immune stimulation triggered by systemic postnatal lipopolysaccharide administration on biochemical, histopathological and morphological measures, which may be relevant to the neurobiology of human psychopathology. In the present study of adult male Wistar rats we examined the brain and plasma levels of monoamines (dopamine, serotonin), their metabolites, the levels of the main excitatory and inhibitory neurotransmitters glutamate and γ-aminobutyric acid and the levels of tryptophan and its metabolites from the kynurenine catabolic pathway. Further, we focused on histopathological and morphological markers related to pathogenesis of brain diseases--glial cell activation, neurodegeneration, hippocampal volume reduction and dopaminergic synthesis in the substantia nigra. Our results show that early immune stimulation in adult animals alters the levels of neurotransmitters and their metabolites, activates the kynurenine pathway of tryptophan metabolism and leads to astrogliosis, hippocampal volume reduction and a decrease of tyrosine hydroxylase immunoreactivity in the substantia nigra. These findings support the crucial pathophysiological role of early immune stimulation in the above mentioned neuropsychiatric disorders.
- MeSH
- astrocyty metabolismus MeSH
- biogenní monoaminy krev metabolismus MeSH
- duševní poruchy imunologie metabolismus patologie psychologie MeSH
- krysa rodu rattus MeSH
- kynurenin metabolismus MeSH
- metabolické sítě a dráhy MeSH
- metabolom MeSH
- mikroglie metabolismus MeSH
- modely nemocí na zvířatech MeSH
- mozek imunologie metabolismus patologie MeSH
- neuroimunomodulace * MeSH
- neurotransmiterové látky metabolismus MeSH
- psychopatologie MeSH
- tělesná hmotnost MeSH
- tryptofan metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
