Two experiments were carried out to examine the impacts of hydroxytyrosol (HT) on lipid metabolism and mitochondrial function in Megalobrama amblycephala. Triplicate groups of fish were fed four test diets: (1) low-fat diet (LFD, 5% fat), (2) high-fat diet (HFD, 15% fat), (3) LFD + 100 mg/kg HT (LFD + HT), and (4) HFD + 100 mg/kg HT (HFD + HT) (in vivo). Hepatocytes from the same batch were exposed to three media including L-15 medium (L15), oleic acid (OA) medium [L15 + 400 μM OA], and OA + HT medium [L15 + 400 μM OA + 10 μM HT] to explore the roles of HT in mitochondrial function (in vitro). Fish fed HFD had excessive fat deposition in the liver, and HT inclusion in the HFD decreased hepatic fat deposition. Transmission electron microscopy revealed that the HFD triggers loss of cristae and metrical density and hydropic changes in mitochondria and that HT supplementation attenuates the ultrastructural alterations of mitochondria. The in vitro test showed that HT decreases fat deposition in hepatocytes, suppresses the reactive oxygen species formation, and facilitates the expression of phospho-AMPK protein and the genes involved in mitochondria biogenesis (PGC-1, NRF-1, TFAM) and autophagy (PINK1, Mul1, Atg5). These findings suggest the lipid-lowering effect of HT mediated by activation of mitochondrial biogenesis and autophagy through the AMPK pathway.
- MeSH
- autofagie * MeSH
- biogeneze organel MeSH
- Cyprinidae genetika metabolismus MeSH
- dietní tuky metabolismus MeSH
- fenethylalkohol analogy a deriváty metabolismus MeSH
- hepatocyty metabolismus MeSH
- játra cytologie metabolismus MeSH
- krmivo pro zvířata analýza MeSH
- mitochondrie metabolismus MeSH
- proteinkinasy aktivované AMP genetika metabolismus MeSH
- rybí proteiny genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The new ultra-high performance liquid chromatography method with tandem mass spectrometry detection (UHPLC-MS/MS) has been optimized to allow fast, selective, and high-throughput analysis of two Candida albicans quorum sensing molecules (QSM), farnesol and tyrosol. The problem of the presence of the interference in the samples and system was successfully solved by careful optimization of chromatographic conditions. Charged hybrid stationary phase modified with pentafluorophenyl group and optimized gradient elution provided adequate separation selectivity and peak shapes. The impurity was identified as dibutyl phthalate and had the same m/z ions as farnesol leading to an important interference on selected reaction monitoring channel. Two different types of biological matrices originating from vaginal fluid, supernatant and sediment, were analysed. Micro-solid phase extraction in pipette tips was optimized for the selective isolation of QSM from the supernatant. The insufficient retention of farnesol on the extraction sorbent was improved when 1% of organic solvent was added prior to extraction, while the retention of tyrosol was only possible when using combined C8 and polymer sorbent type. Strong retention of farnesol had to be solved by increasing elution solvent strength and volume up to 600 μL. However, this approach did not allow the pretreatment of sediment samples due to the sorbent clogging. Therefore, our previously developed protein precipitation method was modified and validated to analyse the sediments. New developed UHPLC-MS/MS method provided suitable accuracy and precision for the determination of QSM in vaginal fluid while using only 50 μL sample volume and two different sample preparation methods.
- MeSH
- Candida albicans izolace a purifikace MeSH
- dospělí MeSH
- farnesol analýza MeSH
- fenethylalkohol analogy a deriváty analýza MeSH
- lidé středního věku MeSH
- lidé MeSH
- limita detekce MeSH
- mladý dospělý MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- vagina chemie mikrobiologie MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- validační studie MeSH
The composition of defensive secretion produced by metathoracic scent glands was analysed in males and females of the milkweed bug Lygaeus equestris (Heteroptera) using gas chromatography with mass spectrometric detection (GC-MS). The bugs were raised either on cardenolide-containing Adonis vernalis or on control sunflower seeds in order to determine whether the possibility to sequester cardenolides from their host plants would affect the composition of defensive scent-gland secretion. Profiles of the composition of defensive secretions of males and females raised on sunflower were closely similar, with predominant presence of (E)-2-octenal, (E)-2-octen-1-ol, decanal and 3-octen-1-ol acetate. The secretion of bugs raised on A. vernalis was more sexually dimorphic, and some chemicals e.g. (E,E)-2,4-hexadienyl acetate and 2-phenylethyl acetate were dominant in males, but absent in females. Compared to bugs from sunflower, the scent-gland secretion of bugs raised on A. vernalis was characterized by lower overall intensity of the peaks obtained for detected chemicals and by absence of some chemicals that have supposedly antipredatory function ((E)-2-hexenal, (E)-4-oxo-hex-2-enal, 2,4-octadienal). The results suggest that there might be a trade-off between the sequestration of defensive chemicals from host plants and their synthesis in metathoracic scent-glands.
- MeSH
- acetáty analýza MeSH
- aldehydy analýza MeSH
- chromatografie plynová MeSH
- čich MeSH
- fenethylalkohol analogy a deriváty analýza MeSH
- feromony analýza MeSH
- Helianthus chemie MeSH
- Heteroptera chemie MeSH
- hlaváček chemie MeSH
- obranné mechanismy proti býložravcům * MeSH
- pachové žlázy chemie MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- pohlavní dimorfismus MeSH
- sexuální faktory MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Potential metabolites of bioactive compounds are important for their biological activities and as authentic standards for metabolic studies. The phenolic compounds contained in olive oil are an important part of the human diet, and therefore their potential metabolites are of utmost interest. We developed a convenient, scalable, one-pot chemoenzymatic method using the arylsulfotransferase from Desulfitobacterium hafniense for the sulfation of the natural olive oil phenols tyrosol, hydroxytyrosol, and of their monoacetylated derivatives. Respective monosulfated (tentative) metabolites were fully structurally characterized using LC-MS, NMR, and HRMS. In addition, Folin-Ciocalteu reduction, 1,1-diphenyl-2-picrylhydrazyl radical scavenging, and antilipoperoxidant activity in rat liver microsomes damaged by tert-butylhydroperoxide were measured and compared to the parent compounds. As expected, the sulfation diminished the radical scavenging properties of the prepared compounds. These compounds will serve as authentic standards of phase II metabolites.
- MeSH
- acetylace MeSH
- arylsulfotransferasa chemie MeSH
- bakteriální proteiny chemie MeSH
- biokatalýza MeSH
- Desulfitobacterium enzymologie MeSH
- fenethylalkohol analogy a deriváty chemická syntéza chemie MeSH
- fenoly chemie MeSH
- hmotnostní spektrometrie MeSH
- molekulární struktura MeSH
- olivový olej chemie MeSH
- scavengery volných radikálů chemická syntéza chemie MeSH
- sírany chemie MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
Velký počet oportunně patogenních bakterií využívá během nástupu infekce systém quorum sensing (QS). Pomocí signálních molekul (u gram -negativních bakterií N -acylhomoserinových laktonů) je řízeno chování celé populace bakterií (hlavně exprese faktorů virulence). Díky studiu QS systémů u gram -negativních bakterií byly již popsány látky se schopností interference s těmito systémy, představující nové léčebné strategie. Téhož by se dalo využít u kvasinek, kde by identifikace QS molekul a pochopení jejich mechanismů působení mohlo vést k efektivnější léčbě kvasinkových infekcí, např. vulvo -vaginální kandidózy způsobované převážně C. albicans. Doposud byly popsány struktury a funkce dvou hlavních signálních molekul účastnících se systému QS u C. albicans, a to farnesolu a tyrosolu. Vzhledem k tomu, že se rezistence mikroorganismů k antibiotikům a dezinfekčním látkám stává stále aktuálnější, je zapotřebí podrobnějšího popisu struktur, funkcí a metabolických drah dalších molekul účastnících se systému QS u kvasinek.
A large number of opportunistic pathogenic bacteria uses quorum sensing (QS) during the beginning of infection process. Signaling molecules (gram -negative bacteria used N -acyl -homoserine lactones) control the behavior of the whole population of bacteria (especially virulence factors expression). Due to the study of QS systems of gram -negative bacteria, compounds which have the ability to interfere with these systems have been described and they represent new treatment strategy. The same approach could be used in the case of yeast, wherein the identification of QS molecules and their mechanism of action could lead to developing much more effective treatment of yeast infections, e.g. vulvo -vaginal candidosis, which is caused mainly by C. albicans. Until now, the structure and function of two main signaling moleculs (farnesol and tyrosol) in the C. albicans QS system were described. Since the resistance of microorganisms to antibiotics and disinfectants becomes more current, detailed description of the structures, functions and the metabolic pathways involved in QS system in yeast is required.
Candida albicans is generally one of the most commonly isolated fungal pathogen from human body. It is a frequent cause of nosocomial infections, bloodstream infections, urinary infections and mucosal infections of oral cavity and vagina C. albicans can grow as hyphae, pseudohyphae, or budding yeast. Morphological conversion of a yeast form to pseudohyphal or hyphal one is often characterized by the change of commensal status to an invasive form. Farnesol and tyrosol can participate in these transformation processes as quorum sensing molecules together with some physical-chemical factors. A new analytical method for identification and quantification of biologically active substances farnesol and tyrosol using ultra high performance liquid chromatography (UHPLC) in connection with tandem mass spectrometry was developed. The analytes were separated on Acquity BEH C18 analytical column using binary mobile phase consisting of acetonitrile and formic acid 0.075% (75:25) at flow-rate 0.20 ml/min. SRM (selected reaction monitoring) mode was applied in order to ensure sufficient selectivity and sensitivity using the first most intensive transition as a quantitative (121>77 and 205>121) and second one for the confirmation purposes (121>93 and 205>109). The method was validated in terms of linearity (>0.9994), precision (0.5-3.8% RSD), accuracy (78.9-106.0%), LOD (limit of detection) and LOQ (limit of quantitation). The method can serve as an analytical tool for the detection and determination of quorum-sensing molecules in biological samples.
- MeSH
- Candida albicans chemie MeSH
- farnesol analýza chemie MeSH
- fenethylalkohol analogy a deriváty analýza chemie MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- kalibrace MeSH
- lidé MeSH
- limita detekce MeSH
- quorum sensing MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cannabis sativa L. plants produce a diverse array of secondary metabolites. Cannabis cell cultures were treated with jasmonic acid (JA) and pectin as elicitors to evaluate their effect on metabolism from two cell lines using NMR spectroscopy and multivariate data analysis. According to principal component analysis (PCA) and partial least square-discriminant analysis (PLS-DA), the chloroform extract of the pectin-treated cultures were more different than control and JA-treated cultures; but in the methanol/water extract the metabolome of the JA-treated cells showed clear differences with control and pectin-treated cultures. Tyrosol, an antioxidant metabolite, was detected in cannabis cell cultures. The tyrosol content increased after eliciting with JA.
- MeSH
- buněčné extrakty chemie izolace a purifikace MeSH
- Cannabis chemie metabolismus MeSH
- cyklopentany metabolismus MeSH
- fenethylalkohol analogy a deriváty analýza MeSH
- kultivované buňky MeSH
- magnetická rezonanční spektroskopie MeSH
- metabolom MeSH
- oxylipiny metabolismus MeSH
- pektiny metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
