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EN
Introduction. Blood biomarkers may support early diagnosis of lung cancer by enabling pre-selection of candidates for computed tomography screening or discrimination between benign and malignant screening-detected nodules. We aimed to identify features of serum metabolome distinguishing individuals with early-detected lung cancer from healthy participants of the lung cancer screening program. Methods. Blood samples were collected in the course of a low-dose computed tomography screening program performed in the Gdansk district (Northern Poland). The analysis included 31 patients with screening-detected lung cancer and the pair-matched group of 92 healthy controls. The gas chromatography coupled to mass spectrometry (GC/MS) approach was used to identify and quantify small metabolites present in serum. Results. There were several metabolites detected in the sera whose abundances discriminated patients with lung cancer from controls. Majority of the differentiating components were downregulated in cancer samples, including amino acids, carboxylic acids and tocopherols, whereas benzaldehyde was the only compound significantly upregulated. A classifier including nine serum metabolites allowed separation of cancer and control samples with 100% sensitivity and 95% specificity. Conclusions. Signature of serum metabolites discriminating between cancer patients and healthy participants of the early lung cancer screening program was identified using a GC/MS metabolomics approach. This signature, though not validated in an independent dataset, deserves further investigation in a larger cohort study.
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2018
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vol. 65
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issue 2
309-318
EN
Several lines of evidence indicate that exposure of heart to ionizing radiation increases the risk of cardiotoxicity manifested by heart dysfunction and cardiovascular diseases. It was initially believed that the heart is an organ relatively resistant to radiation. Currently, however, it is suspected that even low doses of radiation (< 2 Gy) may have a negative impact on the cardiovascular system. Cardiotoxicity of ionizing radiation is associated with metabolic changes observed in cardiac cells injured by radiation. In this study, we used human cardiomyocytes as a model system, and studied their metabolic response to radiation using high-resolution magic angle spinning nuclear magnetic resonance techniques (HR-MAS NMR). Human cardiomyocytes cultured in vitro were exposed to ionizing radiation and their survival was assessed by clonogenic assay. Changes in apoptosis intensity and cell cycle distribution after the irradiation were measured as well. NMR spectra of cardiomyocytes were acquired using Bruker Avance 400 MHz spectrometer at a spinning rate of 3200 Hz. Survival of cardiomyocytes after NMR experiments was assessed by the Trypan blue exclusion assay. Exposure of cardiomyocytes to small doses of ionizing radiation had no effect on cell proliferation potential and intensity of cell death. However, analysis of metabolic profiles revealed changes in lipids, threonine, glycine, glycerophosphocholine, choline, valine, isoleucine, glutamate, reduced glutathione and taurine metabolism. The results of this study showed that ionizing radiation affects metabolic profiles of cardiomyocytes even at low doses, which potentially have no effect on cell viability.
EN
Nowadays, chromatographic methods coupled with mass spectrometry are the most commonly used tools in metabolomics studies. These methods are currently being developed and various techniques and strategies are proposed for the profiling analysis of biological samples. However, the most important thing used to maximize the number of entities in the recorded profiles is the optimization of sample preparation procedure and the data acquisition method. Therefore, ultra high performance liquid chromatography coupled with accurate quadrupoletime- of-flight (Q-TOF) mass spectrometry was used for the comparison of urine metabolomic profiles obtained by the use of various spectral data acquisition methods. The most often used method of registration of metabolomics data acquisition – TOF (MS) was compared with the fast polarity switching MS and auto MS/MS methods with the use of multivariate chemometric analysis (PCA). In all the cases both ionization mode (positive and negative) were studied and the number of the identified compounds was compared. Additionally, various urine sample preparation procedures were tested and it was found that the addition of organic solvents to the sample noticeably reduces the number of entities in the registered profiles. It was also noticed that the auto MS/MS method is the least efficient way to register metabolomic profiles.
EN
Staphylococcus aureus is responsible for many types of infections related to biofilm presence. As the early diagnostics remains the best option for prevention of biofilm infections, the aim of the work presented was to search for differences in metabolite patterns of S. aureus ATCC6538 biofilm vs. free-swimming S. aureus planktonic forms. For this purpose, Nuclear Magnetic Resonance (NMR) spectroscopy was applied. Data obtained were supported by means of Scanning Electron Microscopy, quantitative cultures and X-ray computed microtomography. Metabolic trends accompanying S. aureus biofilm formation were found using Principal Component Analysis (PCA). Levels of isoleucine, alanine and 2,3-butanediol were significantly higher in biofilm than in planktonic forms, whereas level of osmoprotectant glycine-betaine was significantly higher in planktonic forms of S. aureus. Results obtained may find future application in clinical diagnostics of S. aureus biofilm-related infections.
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2017
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vol. 64
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issue 2
215-219
EN
Background: Dengue is one of the major public health problems in the world, affecting more than fifty million cases in tropical and subtropical region every year. The metabolome, as pathophysiological end-points, provide significant understanding of the mechanism and progression of dengue pathogenesis via changes in the metabolite profile of infected patients. Recent developments in diagnostic technologies provide metabolomics for the early detection of infectious diseases. Methods: The mid-stream urine was collected from 96 patients diagnosed with dengue fever at Penang General Hospital (PGH) and 50 healthy volunteers. Urine samples were analyzed with proton nuclear magnetic resonance (1H NMR) spectroscopy, followed by chemometric multivariate analysis. NMR signals highlighted in the orthogonal partial least square-discriminant analysis (OPLS-DA) S-plots were selected and identified using Human Metabolome Database (HMDB) and Chenomx Profiler. A highly predictive model was constructed from urine profile of dengue infected patients versus healthy individuals with the total R2Y (cum) value 0.935, and the total Q2Y (cum) value 0.832. Results: Data showed that dengue infection is related to amino acid metabolism, tricarboxylic acid intermediates cycle and β-oxidation of fatty acids. Distinct variations in certain metabolites were recorded in infected patients including amino acids, various organic acids, betaine, valerylglycine, myo-inositol and glycine. Conclusion: Metabolomics approach provides essential insight into host metabolic disturbances following dengue infection.
EN
Radiotherapy causes molecular changes observed at the level of body fluids, which are potential biomarker candidates for assessment of radiation exposure. Here we analyzed radiotherapy-induced changes in a profile of small metabolites detected in sera of head and neck cancer patients using the gas chromatography coupled with mass spectrometry approach. There were about 20 compounds, including carboxylic acids, sugars, amines and amino acids, whose levels significantly differed between pre-treatment and post-treatment samples. Among metabolites upregulated by radiotherapy there was 3-hydroxybutyric acid, whose level increased about three times in post-treatment samples. Moreover, compounds affected by irradiation were associated with several metabolic pathways, including protein biosynthesis and amino acid metabolism.
EN
Abiotic stresses are produced by inappropriate levels of physical components of the environment and cause plant injury through unique mechanisms that result in specific responses. Metabolomics is a relatively new approach aimed at improved understanding of metabolic networks and the subsequent biochemical composition of plants and other biological organisms. The paper is focused on the use of metabolomics, metabolic profiling and metabolic fingerprinting to study plant responses to some environmental stresses (eg elevated temperature, chilling and freezing, drought, high salinity, UV radiation, high ozone levels, nutrient deficiency, oxidative stress, herbicides and heavy metals). Attention is also devoted to the effects of some environmental factors on plants such as high or low levels of CO2 or different levels of irradiance. Alterations of plants metabolites due to multiple abiotic stresses (drought-heat, drought-salinity, elevated CO2-salinity) are analysed as well. In addition, metabolomic approach to study plant responses to some artificial abiotic stresses, mechanical stress or pulsed electric field-induced stress is discussed. The most important analytical methods applied in metabolomics are presented and perspectives of metabolomics exploitation in the future are outlined, too.
PL
Stres abiotyczny wywołany przez niewłaściwe poziomy fizycznych komponentów środowiska powoduje zmiany w roślinach i poprzez specyficzne mechanizmy prowadzi do określonych odpowiedzi. Metabolomika jest stosunkowo nowym podejściem mającym na celu lepsze zrozumienie szlaków metabolicznych oraz skutków biochemicznych w składzie roślin i innych organizmów biologicznych. Artykuł skupia się na wykorzystaniu metabolomiki, profilowania metabolicznego i "fingerprintingu" metabolicznego do badania reakcji roślin na niektóre stresy środowiskowe (np. podwyższoną temperaturę, chłodzenie i zamrażanie, suszę, wysokie zasolenie, promieniowanie UV, duże stężenie ozonu, niedobór substancji odżywczych, stres oksydacyjny, herbicydy i wpływ metali ciężkich). Zwrócono także uwagę na wpływ niektórych czynników środowiskowych na rośliny, takich jak: wysokie lub niskie poziomy CO2 lub różne poziomy natężenia oświetlenia. Przeanalizowano również zmiany związane z kombinacjami abiotycznych czynników stresujących (susza - upał, susza - zasolenie, podwyższone stężenie CO2 - zasolenie). Omówiono też metabolomiczne podejście do badania reakcji roślin na stresy abiotyczne wywołane niektórymi sztucznymi czynnikami, stresem mechanicznym lub impulsowym polem elektrycznym. Zaprezentowano najważniejsze metody analityczne stosowane w metabolomice oraz nakreślono perspektywy wykorzystania metabolomiki.
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