Ever since proteomics was proven to be capable of characterizing a large number of differences in both protein quality and quantity, it has been applied in various areas of biomedicine, ranging from the deciphering molecular pathogenesis of diseases to the characterization of novel drug targets and the discovery of potential diagnostic biomarkers. Indeed, the biomarker discovery in human plasma is clearly one of the areas with enormous potential. However, without proper planning and implementation of specific techniques, the efforts and expectations may very easily be hampered. Numerous earlier projects aimed at clinical proteomics, characterized by exaggerated enthusiasm, often underestimated some principal obstacles of plasma biomarker discovery. Consequently, ambiguous and insignificant results soon led to a more critical view in this field. In this article, we critically review the current state of proteomic approaches for biomarker discovery and validation, in order to provide basic information and guidelines for both clinicians and researchers. These need to be closely considered prior to initiation of a project aimed at plasma biomarker discovery. We also present a short overview of recent applications of clinical proteomics in biomarker discovery., V. Tambor ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
Dental composite materials often contain monomers with bisphenol A (BPA) structure in their molecules, e.g. bisphenol-A glycidyl dimethacrylate (Bis-GMA). In this study, it was examined whether dental restorative composites could be a low-dose source of BPA or alternative bisphenols, which are known to have endocrine-disrupting effects. Bis-GMA-containing composites Charisma Classic (CC) and Filtek Ultimate Universal Restorative (FU) and “BPA-free” Charisma Diamond (CD) and Admira Fusion (AF) were examined. Specimens (diameter 6 mm, height 2 mm, n=5) were light-cured from one side for 20 s and stored at 37 °C in methanol which was periodically changed over 130 days to determine the kinetics of BPA release. BPA concentrations were measured using a dansyl chloride derivatization method with liquid chromatography - tandem mass spectrometry detection. The amounts of BPA were expressed in nanograms per gram of composite (ng/g). BPA release from Bis-GMA-containing CC and FU was significantly higher compared to “BPA-free” CD and AF. The highest 1-day release was detected with FU (15.4±0.8 ng/g), followed by CC (9.1±1.1 ng/g), AF (2.1±1.3 ng/g), and CD (1.6±0.8 ng/g), and the release gradually decreased over the examined period. Detected values were several orders of magnitude below the tolerable daily intake (4 µg/kg body weight/day). Alternative bisphenols were not detected. BPA was released even from “BPA-free” composites, although in significantly lower amounts than from Bis-GMA-containing composites. Despite incubation in methanol, detected amounts of BPA were substantially lower than current limits suggesting that dental composites should not pose a health risk if adequately polymerized., Markéta Šimková, Antonín Tichý, Michaela Dušková, Pavel Bradna., and Obsahuje bibliografii
Příspěvek poskytuje přehlednou informaci o základních fyzikálních principech, na nichž pracují současné hmotnostní spektrometry, a informaci o základních principech některých důležitých soudobých hmotnostně spektrálních metod v biologickém výzkumu., Jiří Knížek, Zdeněk Půlpán, Martin Hubálek, Ladislav Beránek, Petr Pokorný., and Obsahuje seznam literatury
The present study was undertaken to identify potentially immunoreactive proteins of the muscle larvae (ML) and adult stage (Ad) of the nematode Trichinella spiralis Owen, 1835. To identify immunoreactive proteins that are specifically recognised by anti-Trichinella antibodies, ML and Ad crude extracts and their excretory-secretory (E-S) products were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot with serum samples from pigs experimentally infected with T. spiralis. A total of 18 bands were selected for final identification by liquid chromatography-tandem mass spectrometry. To further understand the functions of the proteins identified in this study, gene ontology terms were applied. Results showed that the specific antibodies against T. spiralis reacted with protein bands matching heat shock proteins, aminopeptidase, enolase, isocitrate dehydrogenase NADP-dependent, tropomyosin, P49 antigen, serine proteinase, secreted 5'-nucleotidase, antigen targeted by protective antibodies, 53 kDa E-S antigen, putative trypsin and paramyosin. Three proteins common for both adult stage and muscle larvae, including heat shock proteins, enolase and 5'-nucleotidase, might play important role during T. spiralis infection. These proteins are presumably presented to the host immune system and may induce humoral immune response. Thus, these proteins may be potential antigens for early diagnosis and the development of a vaccine against the parasite., Justyna Bien, Wladyslaw Cabaj, Bozena Moskwa., and Obsahuje bibliografii
In this study, we presented a new approach for quantification of bicarbonate (HCO3-) molecules bound to PSII. Our method, which is based on a combination of membrane-inlet mass spectrometry (MIMS) and 18O-labelling, excludes the possibility of "non-accounted" HCO3- by avoiding (1) the employment of formate for removal of HCO3- from PSII, and (2) the extremely low concentrations of HCO3-/CO2 during online MIMS measurements. By equilibration of PSII sample to ambient CO2 concentration of dissolved CO2/HCO3-, the method ensures that all physiological binding sites are saturated before analysis. With this approach, we determined that in spinach PSII membrane fragments 1.1 ± 0.1 HCO3- are bound per PSII reaction center, while none was bound to isolated PsbO protein. Our present results confirmed that PSII binds one HCO3- molecule as ligand to the non-heme iron of PSII, while unbound HCO3- optimizes the water-splitting reactions by acting as a mobile proton shuttle., K. Tikhonov, D. Shevela, V. V. Klimov, J. Messinger., and Obsahuje bibliografické odkazy
The presence of adipokinetic activity in crude extracts of corpora cardiaca (CC) from the butterfly (Vanessa cardui L., Nymphalidae) was demonstrated by bioassay and Mas-AKH was revealed as the major adipokinetic hormone (AKH) by use of two different technologies of sequence elucidation: HPLC separation of the peptide followed by Edman degradation and Q-TOF mass spectrometry. In contrast to the time- and material-consuming conventional methods of peptide purification and sequencing, substantial structural data of the peptide were confirmed - post factum - from one pCC (pair of CC) by Q-TOF mass spectrometry. Only males of our laboratory colony showed a significant lipid increase in the haemolymph after injection of either crude CC extract (1 pCC equivalent) or 10 pmol of synthetic peptide., Gabriele Köllisch, Peter D. Verhaert, Matthias W. Lorenz, Roland Kellner, Gerd Gäde, Klaus H. Hoffmann, and Lit