Wall paintings are integral to cultural heritage and offer rich insights into historical and religious beliefs. There exist various wall painting techniques that pose challenges in binder and pigment identification, especially in the case of egg/oil-based binders. GC-MS identification of lipidic binders relies routinely on parameters like the ratios of fatty acids within the plaster. However, the reliability of these ratios for binder identification is severely limited, as demonstrated in this manuscript. Therefore, a more reliable tool for effective differentiation between egg and oil binders based on a combination of diagnostic values, specific markers (cholesterol oxidation products), and PCA is presented in this study. Reference samples of wall paintings with egg and linseed oil binders with six different pigments were subjected to modern artificial ageing methods and subsequently analysed using two GC-MS instruments. A statistically significant difference (at a 95% confidence level) between the egg and oil binders and between the results from two GC-MS instruments was observed. These discrepancies between the results from the two GC-MS instruments are likely attributed to the heterogeneity of the samples with egg and oil binders. This study highlights the complexities in identifying wall painting binders and the need for innovative and revised analytical methods in conservation efforts.
Cancer cells depend on nucleotides for proliferation. Inhibition of nucleotide metabolism by antimetabolites is a well-established anticancer therapy. However, resistance and toxicity to antimetabolite treatments reduce their effectiveness. Here, we focus on the pyrimidine de novo synthesis pathway, which is crucial for cancer cell proliferation, yet its pharmacological targeting in cancer has been without much clinical success so far. Hence, it is important to understand how cancer cells cope with the insufficiency of this pathway. Here, we describe a procedure to prepare subcutaneous tumor model deficient in de novo pyrimidine synthesis. For examination of metabolic responses to de novo synthesis blockade in tumors, we propose application of MALDI imaging that allows spatially resolved examination of metabolic responses to de novo synthesis blockade in tumors.
The ability to adapt to environmental stress, including therapeutic insult, contributes to tumor evolution and drug resistance. In suboptimal conditions, the integrated stress response (ISR) promotes survival by dampening cytosolic translation. We show that ISR-dependent survival also relies on a concomitant up-regulation of mitochondrial protein synthesis, a vulnerability that can be exploited using mitoribosome-targeting antibiotics. Accordingly, such agents sensitized to MAPK inhibition, thus preventing the development of resistance in BRAFV600E melanoma models. Additionally, this treatment compromised the growth of melanomas that exhibited elevated ISR activity and resistance to both immunotherapy and targeted therapy. In keeping with this, pharmacological inactivation of ISR, or silencing of ATF4, rescued the antitumoral response to the tetracyclines. Moreover, a melanoma patient exposed to doxycycline experienced complete and long-lasting response of a treatment-resistant lesion. Our study indicates that the repurposing of mitoribosome-targeting antibiotics offers a rational salvage strategy for targeted therapy in BRAF mutant melanoma and a therapeutic option for NRAS-driven and immunotherapy-resistant tumors.
- MeSH
- chemorezistence účinky léků MeSH
- doxycyklin farmakologie MeSH
- fyziologický stres účinky léků MeSH
- inhibitory proteinkinas farmakologie MeSH
- lidé MeSH
- melanom farmakoterapie genetika mortalita patologie MeSH
- mitochondriální ribozomy účinky léků MeSH
- myši inbrední C57BL MeSH
- myši nahé MeSH
- nádorové buněčné linie MeSH
- nádory uvey farmakoterapie patologie MeSH
- protinádorová antibiotika farmakologie MeSH
- senioři MeSH
- tigecyklin farmakologie MeSH
- xenogenní modely - testy protinádorové aktivity MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- práce podpořená grantem MeSH
- Klíčová slova
- Mispregnol©,
- MeSH
- aplikace intravaginální MeSH
- cervix uteri * účinky léků MeSH
- dilatace a kyretáž metody MeSH
- hysteroskopie * metody MeSH
- klinická studie jako téma MeSH
- lidé MeSH
- misoprostol aplikace a dávkování škodlivé účinky terapeutické užití MeSH
- postmenopauza MeSH
- předoperační péče metody MeSH
- premedikace * metody MeSH
- premenopauza MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- přehledy MeSH
In this study, a reproducible method of fabricating hierarchically 3D porous scaffolds with high porosity and pore interconnectivity is reported. The method is based on in-situ foaming of a dispersion of diisocyanate, polyol, water and hydroxyapatite (HA) to form a hard foamed HA/polyurethane composite which after heat treatment provided a bi-phase calcium phosphate scaffold. This technique, combining the advantages of polymer sponge and direct foaming methods, provides a better control over the macrostructure of the scaffold. A modification of the multi-scaled porous macrostructure of scaffolds produced by changing the ratio of input reactants and by sintering temperature was studied. The pore morphology, size, and distribution were characterized using a scanning electron microscope and mercury porosimetry. The pores were open and interconnected with multi-scale (from several nanometres to millimetres) sizes convenient for using in tissue engineering applications. The bioactivity was confirmed by growing an apatite layer on the surfaces after immersion in simulated body fluid. The material was biocompatible, as shown by using normal human adipose tissue-derived stem cells (ASC). When seeded onto the scaffolds, the ASC adhered and remained healthy while maintaining their typical morphology.
The stability of in vitro cell cultures is an important issue for any clinical, bio-industrial, or pharmacological use. Embryonic stem cells are pluripotent; consequently, they possess the ability to differentiate into all three germ layers and are inherently prone to respond to differentiation stimuli. However, long-term culture inevitably yields clones that are best adapted to the culture conditions, passaging regimes, or differentiation sensitivity. This cellular plasticity is a major obstacle in the development of bio-industrial or clinical-grade cultures. At present, the quality control of cell cultures is limited by the lack of reliable (epi)genetic or molecular markers or by the focus on a particular type of instability such as karyotype abnormalities or adverse phenotypic traits. Therefore, there is an ongoing need for robust, feasible, and sensitive methods of determining or confirming cell status and for revealing potential divergences from the optimal state. We modeled both intrinsic and extrinsic changes in human embryonic stem cell (hESC) states using different experimental strategies and addressed the changes in cell status by intact cell mass spectrometry fingerprinting. The analysis of spectral fingerprints by methods routinely used in analytical chemistry clearly distinguished the morphologically and biochemically similar populations of hESCs and provided a biomarker-independent tool for the quality control of cell culture. Stem Cells Translational Medicine 2018;7:109-114.
- MeSH
- biologické markery analýza MeSH
- buněčné kultury MeSH
- fenotyp MeSH
- kultivované buňky MeSH
- lidé MeSH
- lidské embryonální kmenové buňky fyziologie MeSH
- plasticita buňky fyziologie MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In this study, a soil from two ceramic vessels belonging to Corded Ware culture, 2707⁻2571 B.C., found in a cremation grave discovered in Central Moravia, Czech Republic, was analyzed using matrix-assisted laser desorption/ionization⁻mass spectrometry (MALDI⁻MS) combined with advanced statistical treatment (principal component analysis, PCA, and orthogonal projection to latent structures discriminant analysis, OPLS-DA) and by enzyme-linked immunosorbent assay (ELISA). MALDI⁻MS revealed the presence of triacylglycerols in both vessels. This analytical technique was used for the analysis of the soil content from archaeological ceramic vessels for the first time. Targeted ELISA experiments consequently proved the presence of milk proteins in both ceramic vessels. These results represent the first direct evidence of the use of milk or dairy products in the Eneolithic period in Moravian Corded Ware Culture and help to better understand the diet habits and living conditions of Eneolithic populations in Central Europe.
A microscale discontinuous fermenter was used for anaerobic digestion of wine waste - a hardly gasifiable feedstock material. Efficiency of biogas production, i.e. changes in content of nitrogen, oxygen, carbon dioxide and methane in gas phase, was monitored by gas chromatography/mass spectrometry. Liquid chromatography/high resolution tandem mass spectrometry in combination with principal component analysis and orthogonal projection to latent structures was used to reveal main chemical differences of gasified wine waste mixture from commonly used ones in agricultural biogas plants. Compounds with particular polyphenolic structures appeared among the most distinctive markers. Analysis of samples collected during acidogenic phase and unstabilized methanogenesis indicates formation of certain dihydro-flavonoids in early stages of the process and their consequent degradation. Due to formerly described higher toxicity of some dihydroflavonoids (e.g. taxifolin) compared to their more common counterparts (e.g. quercetin, malvidin etc.), unstabilized digestate would represent a potential environmental risk when used as a fertilizer deserving a proper control.
- MeSH
- anaerobióza MeSH
- biopaliva analýza MeSH
- bioreaktory MeSH
- chromatografie kapalinová metody MeSH
- dusík analýza MeSH
- methan analýza MeSH
- odpadky - odstraňování metody MeSH
- polyfenoly chemie metabolismus MeSH
- potravinářský průmysl MeSH
- průmyslová hnojiva analýza MeSH
- průmyslový odpad MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- Vitis chemie MeSH
- Publikační typ
- časopisecké články MeSH
Condensation of anthocyanins and their aglycons with small organic molecules yields more stable natural dyes, e.g. pyranoanthocyanins arising spontaneously in various food products. Reaction of pelargonidin with acetone produces two isomeric anthocyanidin dyes - 5-methylpyranopelargonidin and 4-methylfuropelargonidin. A robust semipreparative liquid chromatographic method was developed to isolate both derivatives from a simple aged solution of pelargonidin in methanol: acetone: 37% aqueous hydrochloric acid (1:1:0.008, v/v/v). Detailed interpretation of mass and nuclear magnetic resonance spectra allowed to assign structures of both dyes in isolated fractions. A fast UHPLC-MS method was optimized for the control of their production in the reaction mixture. Use of reversed stationary phase and acidic mobile phases in gradient mode allowed separation of both isomers in less than one minute. Fragmentation of both dyes after collision activated dissociation in collision cell was studied comprehensively and the observed processes were compared with data from quantum calculations (computational chemistry utilizing DFT methods). When comparing both isomers, retro-Diels-Alder fragmentation appears preferred in furo-derivative, while small losses (i.e. methane and water molecules) were more pronounced in pyrano-derivative. Both studied isomeric dyes were found in laboratory prepared strawberry fruit wine and their content was compared with major present anthocyanins and their derivatives.
- MeSH
- anthokyaniny analýza chemická syntéza izolace a purifikace MeSH
- barvicí látky analýza chemická syntéza izolace a purifikace MeSH
- chemické techniky analytické metody MeSH
- jahodník * chemie MeSH
- ovoce chemie MeSH
- víno analýza MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
Nano-liquid chromatography and conventional HPLC were used for the separation of diastereomers of (+)-catechin-ethyl-malvidin-3-glucoside. Those bridged anthocyanin dyes were obtained by reaction of (+)-catechin with malvidin-3-glucoside in the presence of acetaldehyde. Both diastereomers were isolated with semipreparative chromatography and their structures were confirmed by nuclear magnetic resonance and mass spectrometry. In-laboratory prepared capillary columns packed with fully porous particles Chromosphere C18, dp=3μm, core-shell particles Kinetex C18, dp=2.6μm (100μm i.d.) and monolithic column Chromolith CapRod (100μm i.d.) were used for the separation of (+)-catechin, malvidin-3-glucoside and both diastereomers. Chromosphere C18 stationary phase provided the best chromatographic performance. Mobile phase containing water:acetonitrile (80:20) acidified with trifluoroacetic acid (0.1%, v/v/v) was used in an isocratic elution mode with a flow rate of 360nLmin(-1). Separation of studied compounds was achieved in less than 7min under optimized conditions. The nano-liquid chromatographic method and a conventional HPLC one using the same fully porous particles (Chromosphere C18, 3μm, 100mm×4.6mm) were compared providing higher separation efficiency with the first analytical method and similar selectivity. A better peak symmetry and higher resolution of the studied diastereomers was achieved by conventional chromatography. Nevertheless, nano-liquid chromatography appeared to be useful for the separation of complex anthocyanin dyes and can be utilized for their analysis in plant and food micro-samples. The developed method was used for analysis of red wine grape pomace.
- MeSH
- acetaldehyd chemie MeSH
- acetonitrily chemie MeSH
- analýza potravin metody MeSH
- anthokyaniny chemie MeSH
- glukosidy chemie MeSH
- hmotnostní spektrometrie MeSH
- katechin chemie MeSH
- poréznost MeSH
- Vitis chemie MeSH
- vysokoúčinná kapalinová chromatografie * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
