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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.
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.
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.
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.
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.
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.
31P high resolution nuclear magnetic resonance (NMR) spectroscopy was used to examine phospholipid metabolism and to analyze the phosphate-containing compounds in the bile in the transplanted liver recipients, the cholelithiasis patients' and the living donors' groups. Three signals of NMR spectrum of raw bile were determined: inorganic phosphate (Pi), lysophosphatidylcholine (LPtdC), and phosphatidylcholine (PtdC) in all investigated groups. Pi concentration was significantly higher in the recipients' group than in the living donors' group (Mann-Whitney test, p < 0.05). LPtdC and PtdC concentrations were significantly higher (Mann-Whitney test, p < 0.05) in the cholelithiasis patients' group in comparison to the recipients' group. Between the cholelithiasis patients' group and the living donors' group no significant differences in the three analysed compounds were found. The chemometric analysis for the 31P NMR spectral data set provided good classifications between the living donors' and recipients' groups and the poor one among all groups. Results of our study suggest that 31P NMR spectroscopy in vitro may be used for assessment of graft function, for the early signs of rejection and for the predisposition to gallstone formation.
In this review, we examine the state-of-the-art technologies (gas and liquid chromatography, mass spectroscopy and nuclear magnetic resonance, etc.) in the well-established area of metabolomics especially as they relate to protozoan parasites.
Metabolomics approaches allow systematic identification and quantitation of all metabolites in biological samples analyzes. As already known metabolism is directly or indirectly related to every aspect of cell function, therefore a careful observation of every changes taking place in metabolism, for example endogenous biochemical reaction products, reflectsthe phenotype of any living cell. Monitoring the metabolite profiles using metabolomics technologies, especially nuclear magnetic resonance (NMR) spectroscopy based on cell culture, allows us to evaluate drug efficiency and outcome of experimental therapy, and most importantly, it allows us to monitor the reaction of the model cell lines to a given treatment. The continued development of metabolomic approaches, e.g. analytical technique, or chemometric software, will accelerate the widespread use of metabolomics not only in the clinical field but also in different biological fields. This work presents a use of nuclear magnetic resonance to characterize and understand the cellular metabolome in a wide range of pathophysiological and clinical contexts.
Artykuł powstał na podstawie rozprawy doktorskiej pt.: 'The application of novel analytical techniques for metabolomics analysis of human breast milk samples' nagrodzonej przez Komitet chemii Analitycznej PAN w 2021 roku w konkursie na najlepsze prace doktorskie, nagroda ufundowana przez firmę Anchem.
A heteropolysaccharide, named L2, from Lentinula edodes has been proved to possess immunostimulating and anti-ageing activities in previous studies, but its acting mechanism was not completely understood. In this study, 1 H NMR spectroscopy approach was employed to investigate the metabolic profi les of the urine from adult mice after L2 intervention. Using principal component analysis (PCA) and partial least squares-discriminate analysis (PLS-DA), 22 potential biomarkers were found to be mainly involved in some metabolic pathways: amino acid metabolism, energy metabolism, lipid metabolism, tricarboxylic acid (TCA) cycle, urea cycle and gut microbiota metabolism. Among them, the signifi cantly altered metabolites include: elevated glutamate (75%) and creatine (64%); decreased proline (65%), betaine (58%), fucose (63%) and dimethylamine (59%). In conclusion, the present data is helpful to understand the mechanisms related to previously confi rmed immunomodulation and anti-aging effects of L2, and provide valuable information for mining new functions of L2.
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