Cisplatin (cis-diamminedichloroplatinum II; CDDP) is a widely used cytostatic agent; however, it tends to promote kidney and liver disease, which are a major signs of drug-induced toxicity. Platinum compounds are often presented as alternative therapeutics and subsequently easily dispersed in the environment as contaminants. Due to the major roles of the liver and kidneys in removing toxic materials from the human body, we performed a comparative study of the amino acid profiles in chicken liver and kidneys before and after the application of CDDP and platinum nanoparticles (PtNPs-10 and PtNPs-40). The treatment of the liver with the selected drugs affected different amino acids; however, Leu and Arg were decreased after all treatments. The treatment of the kidneys with CDDP mostly affected Val; PtNPs-10 decreased Val, Ile and Thr; and PtNPs-40 affected only Pro. In addition, we tested the same drugs on two healthy cell lines, HaCaT and HEK-293, and ultimately explored the amino acid profiles in relation to the tricarboxylic acid cycle (TCA) and methionine cycle, which revealed that in both cell lines, there was a general increase in amino acid concentrations associated with changes in the concentrations of the metabolites of these cycles.
- Publikační typ
- časopisecké články MeSH
The coronavirus pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) directly or indirectly affects every individual worldwide. The fight against SARS-CoV-2 is based on the rapid and accurate diagnosis and subsequent isolation of infected individuals. Therefore, the demands for the scientific development of diagnostic methods for the confirmation of SARS-CoV-2 are enormous. Currently, reverse-transcription quantitative polymerase chain reaction (RT-qPCR) is the main method used for detecting viruses, including SARS-CoV-2, and is considered the gold standard for coronavirus disease 2019 (COVID-19) identification. However, various alternatives have been investigated due to the time and cost demands of this method or to shortages of reagents. In this review, we focus on matrix-assisted laser desorption and ionisation with time-of-flight analyser mass spectrometry (MALDI-TOF MS) techniques as potential tools for the diagnosis of viruses with an emphasis on SARS-CoV-2. MALDI-TOF is commonly used in clinical laboratories for bacterial characterization and identification, but in the field of clinical virology, MALDI-TOF remains only a promising technology for routine diagnosis. This review provides an overview of the development of clinical virology from the point of view of using MALDI-TOF for virus identification and as a possible diagnostic tool for SARS-CoV-2 detection. In addition, this review summarizes the current state of standard methods for virus diagnostics including the preparation of clinical samples.
- MeSH
- COVID-19 * diagnóza MeSH
- lidé MeSH
- pandemie MeSH
- SARS-CoV-2 MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- viry * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Here we developed a powerful tool for comprehensive data collection and mapping of molecular and elemental signatures in the Melanoma-bearing Libechov Minipig (MeLiM) model. The combination of different mass spectrometric methods allowed for detail investigation of specific melanoma markers and elements and their spatial distribution in tissue sections. MALDI-MSI combined with HPLC-MS/MS analyses resulted in identification of seven specific proteins, S100A12, CD163, MMP-2, galectin-1, tenascin, resistin and PCNA that were presented in the melanoma signatures. Furthermore, the ICP-MS method allowed for spatial detection of zinc, calcium, copper, and iron elements linked with the allocation of the specific binding proteins.
The here presented work is focused on the development of a method for detection of microbial contamination of food based on uracil-selective synthetic receptors. Because uracil may serve as an indicator of bacterial contamination, its selective and on-site detection may prevent spreading of foodborne diseases. The synthetic receptors were created by molecular imprinting. Molecularly imprinted polymers for selective uracil isolation were prepared by a non-covalent imprinting method using dopamine as a functional monomer. Detection of isolated uracil was performed by capillary electrophoresis with absorption detection (λ - 260 nm). The conditions of preparation of molecularly imprinted polymers, their binding properties, adsorption kinetics and selectivity were investigated in detail. Furthermore, the prepared polymer materials were used for selective isolation and detection of uracil from complex samples as tomato products by miniaturized electrophoretic system suggesting the potential of in situ analysis of real samples.
- MeSH
- adsorpce MeSH
- molekulový imprinting * MeSH
- polymery MeSH
- receptory umělé * MeSH
- uracil MeSH
- Publikační typ
- časopisecké články MeSH
Identification of metal-binding sites in proteins and understanding metal-coupled protein folding mechanisms are aspects of high importance for the structure-to-function relationship. Mass spectrometry (MS) has brought a powerful adjunct perspective to structural biology, obtaining from metal-to-protein stoichiometry to quaternary structure information. Currently, the different experimental and/or instrumental setups usually require the use of multiple data analysis software, and in some cases, they lack some of the main data analysis steps (MS processing, scoring, identification). Here, we present a comprehensive data analysis pipeline that addresses charge-state deconvolution, statistical scoring, and mass assignment for native MS, bottom-up, and native top-down with emphasis on metal-protein complexes. We have evaluated all of the approaches using assemblies of increasing complexity, including free and chemically labeled proteins, from low- to high-resolution MS. In all cases, the results have been compared with common software and proved how MetaOdysseus outperformed them.
- MeSH
- COVID-19 diagnóza virologie MeSH
- design vybavení MeSH
- lidé MeSH
- point of care testing MeSH
- RNA virová analýza genetika MeSH
- SARS-CoV-2 genetika izolace a purifikace MeSH
- techniky amplifikace nukleových kyselin přístrojové vybavení MeSH
- testování na COVID-19 průkazem nukleové kyseliny přístrojové vybavení MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- úvodníky MeSH
Coronavirus disease (COVID-19) caused by SARS-CoV-2 has spread since the end of 2019 and has resulted in a pandemic with unprecedented socioeconomic consequences. This situation has created enormous demand for the improvement of current diagnostic methods and the development of new diagnostic methods for fast, low-cost and user-friendly confirmation of SARS-CoV-2 infection. This critical review focuses on viral electrochemical biosensors that are promising for the development of rapid medical COVID-19 diagnostic tools. The molecular biological properties of SARS-CoV-2 as well as currently known biochemical attributes of infection necessary for biosensor development are outlined. The advantages and drawbacks of conventional diagnostic methods, such as quantitative reverse-transcription polymerase chain reaction (qRT-PCR), are critically discussed. Electrochemical biosensors focusing on viral nucleic acid and whole viral particle detection are highlighted and discussed in detail. Finally, future perspectives on viral electrochemical biosensor development are briefly mentioned.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Publikační typ
- abstrakt z konference MeSH
Respiratory infections are a real threat for humans, and therefore the pig model is of interest for studies. As one of a case for studies, Actinobacillus pleuropneumoniae (APP) caused infections and still worries many pig breeders around the world. To better understand the influence of pathogenic effect of APP on a respiratory system-lungs and tracheobronchial lymph nodes (TBLN), we aimed to employ matrix-assisted laser desorption/ionization time-of-flight mass spectrometry imaging (MALDI-TOF MSI). In this study, six pigs were intranasally infected by APP and two were used as non-infected control, and 48 cryosections have been obtained. MALDI-TOF MSI and immunohistochemistry (IHC) were used to study spatial distribution of infectious markers, especially interleukins, in cryosections of porcine tissues of lungs (necrotic area, marginal zone) and tracheobronchial lymph nodes (TBLN) from pigs infected by APP. CD163, interleukin 1β (IL‑1β) and a protegrin-4 precursor were successfully detected based on their tryptic fragments. CD163 and IL‑1β were confirmed also by IHC. The protegrin-4 precursor was identified by MALDI-TOF/TOF directly on the tissue cryosections. CD163, IL‑1β and protegrin‑4 precursor were all significantly (p < 0.001) more expressed in necrotic areas of lungs infected by APP than in marginal zone, TBLN and in control lungs.
- MeSH
- Actinobacillus pleuropneumoniae patogenita MeSH
- antigeny diferenciační myelomonocytární metabolismus MeSH
- biologické markery metabolismus MeSH
- bronchy metabolismus MeSH
- CD antigeny metabolismus MeSH
- infekce bakteriemi rodu Actinobacillus metabolismus mikrobiologie MeSH
- infekce dýchací soustavy metabolismus mikrobiologie MeSH
- interleukin-1beta metabolismus MeSH
- kationické antimikrobiální peptidy metabolismus MeSH
- lymfatické uzliny metabolismus MeSH
- plíce metabolismus MeSH
- prasata MeSH
- receptory buněčného povrchu metabolismus MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Dietary supplementation with polyunsaturated fatty acids (PUFA) n-3 can affect cutaneous wound healing; however, recent findings demonstrate the variable extent of their influence on the quality of healing. Here, we compare the effect of several dietary oils, containing different levels of PUFA n-3 and PUFA n-6, on wound healing in the rat model. Rats were fed the feed mixture with 8% palm oil (P), safflower oil (S), fish oil (F) or Schizochytrium microalga extract (Sch) and compared to the animals fed by control feed mixture (C). Dorsal full-thickness cutaneous excisions were performed after 52 days of feeding and skin was left to heal for an additional 12 days. Histopathological analysis of skin wounds was performed, including immune cells immunolabeling and the determination of hydroxyproline amount as well as gene expression analyses of molecules contributing to different steps of the healing. Matrix-assisted-laser-desorption-ionization mass-spectrometry-imaging (MALDI-MSI) was used to determine the amount of collagen α-1(III) chain fragment in healing samples. Treatment by Schizochytrium extract resulted in decrease in the total wound area, in contrast to the safflower oil group where the size of the wound was larger when comparing to control animals. Diet with Schizochytrium extract and safflower oils displayed a tendency to increase the number of new vessels. The number of MPO-positive cells was diminished following any of oil treatment in comparison to the control, but their highest amount was found in animals with a fish oil diet. On the other hand, the number of CD68-positive macrophages was increased, with the most significant enhancement in the fish oil and safflower oil group. Hydroxyproline concentration was the highest in the safflower oil group but it was also enhanced in all other analyzed treatments in comparison to the control. MALDI-MSI signal intensity of a collagen III fragment decreased in the sequence C > S > Sch > P > F treatment. In conclusion, we observed differences in tissue response during healing between dietary oils, with the activation of inflammation observed following the treatment with oil containing high eicosapentaenoic acid (EPA) level (fish oil) and enhanced healing features were induced by the diet with high content of docosahexaenoic acid (DHA, Schizochytrium extract).
- MeSH
- antigeny CD8 metabolismus MeSH
- dietní tuky nenasycené aplikace a dávkování farmakologie MeSH
- hojení ran účinky léků MeSH
- indoly chemie MeSH
- kolagen typ III metabolismus MeSH
- krysa rodu rattus MeSH
- kůže účinky léků zranění metabolismus MeSH
- kyseliny mastné omega-3 analýza MeSH
- kyseliny mastné omega-6 analýza MeSH
- makrofágy imunologie MeSH
- modely nemocí na zvířatech MeSH
- palmový olej aplikace a dávkování chemie farmakologie MeSH
- rybí oleje aplikace a dávkování chemie farmakologie MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- světlicový olej aplikace a dávkování chemie farmakologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
