Předmětem řešení je identifikace plazmatických proteinů, jejíchž stanovování v klinické praxi by v budoucnu mohlo vést k jednoznačné diagnostice hypertrofické kardiomypatie.; The aim of the project is detection and identification of new plasma biomarkers that could be used for the unambiguous diagnosis of hypertrophic cardiomyopathy in the clinical praxis in the future.

• MeSH
• biologické markery analýza   MeSH
• hypertrofická kardiomyopatie diagnóza   patofyziologie   MeSH
• kardiomyocyty patologie   MeSH
• krevní proteiny analýza   MeSH
• myofibrily patologie   MeSH
• proteomika metody   MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• kardiologie
• biologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

It is usual for information to be unavailable regarding the molecular composition of extracts from herbs or animal tissues that are popular in folk medicine. Here, we present analysis of the alcohol-ether extract from bovine tissue analogous to the basic substance used in such commercial products as Retisin, Imuregen, Actovegin, and Solcoseryl. The tested extract contains a whole spectrum of free amino acids, small proteins and oligopeptides of molecular weight up to 10 kDa, various nucleotides, and a small amount of phospholipids. Among the molecules that can explain some biological activities of the extract were identified those of taurine (2-aminoethanesulfonic acid, a derivative of the amino acid cysteine), several defensins, and bactericidal hemoglobin fragments known as hemocidins. All those molecules identified are natural components of bovine tissues, and a substantial number of them might be biologically active in vivo. Others are sources of readily available nutrients.

• Klíčová slova
• Juvenil,
• MeSH
• lidé MeSH
• potravní doplňky analýza   MeSH
• tkáňové extrakty MeSH
• Check Tag
• lidé MeSH

• Publikační typ
• abstrakt z konference MeSH

One of the key requirements of the biodefense system of the Czech Armed Forces is a capability to identify the biological warfare agents (BWA). In this regard the Central Military Health Institute that is responsible for the biodefense in the Czech Armed Forces took part in 10thAnnual international comparative exercise of the NATO (North Atlantic Treaty Organization) military biological laboratories. The aim of this test was the identification of Bacillus anthracis in unknown samples which were contaminated by different disinfectants. Real-time polymerase chain reaction (PCR) was chosen as optimal method for this exercise because of a robustness, speed and flexibility of this method. Due to the presence of a disinfectant the identification procedure could only be conducted after including an additional step to sample preparation. Tandem mass spectrometry was selected as a confirmatory method for the exercise. Our test result was in full agreement with the exercise design. This exercise confirmed that method of provisional identification deployable in mobile component of biodefense system of the Czech Armed Forces is sensitive and robust for use in the field conditions. The tandem mass spectrometry analysis confirmed PCR results and verified the valuable confirmatory role of mass spectrometry in the identification of biological agents.

• MeSH
• Bacillus anthracis genetika   izolace a purifikace   MeSH
• bakteriologické techniky metody   přístrojové vybavení   využití   MeSH
• biologické bojové látky MeSH
• DNA bakterií analýza   genetika   MeSH
• financování organizované MeSH
• hodnotící studie jako téma MeSH
• laboratoře MeSH
• lidé MeSH
• nebezpečné látky izolace a purifikace   MeSH
• polymerázová řetězová reakce metody   přístrojové vybavení   využití   MeSH
• senzitivita a specificita MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• vojenská věda MeSH
• Check Tag
• lidé MeSH

Pathogenic bacteria have developed various mechanisms to evade host immune defense systems. Invasion of pathogenic bacteria requires interaction of the pathogen with host receptors, followed by activation of signal transduction pathways and rearrangement of the cytoskeleton to facilitate bacterial entry. Numerous bacteria exploit specialized plasma membrane microdomains, commonly called membrane rafts, which are rich in cholesterol, sphingolipids and a special set of signaling molecules which allow entry to host cells and establishment of a protected niche within the host. This review focuses on the current understanding of the raft hypothesis and the means by which pathogenic bacteria subvert membrane microdomains to promote infection.

• MeSH
• Bacteria patogenita   MeSH
• bakteriální infekce mikrobiologie   MeSH
• interakce hostitele a patogenu MeSH
• lidé MeSH
• membránové mikrodomény mikrobiologie   fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

BACKGROUND: Both BALB/c mice and common voles (Microtus arvalis) are considered highly susceptible to tularemia. However, the common vole is reported to harbour Francisella tularensis in European habitats as well as to survive longer with chronic shedding of the bacterium. The purpose of the present study was to compare the response of these two rodents to a wild Francisella tularensis subsp. holarctica strain infection. METHODS: Rodents were evaluated for differences in the total antioxidant capacity derived from low-molecular-weight antioxidants, biochemistry including lipid metabolism, tissue bacterial burdens and histopathology following experimental intraperitoneal infection with 160 colony forming units (CFU) pro toto. RESULTS: Bacterial burdens in common voles started to develop later post-exposure and amounted to lower levels than in BALB/c mice. Elevation of liver function enzymes was more pronounced in mice than common voles and there were marked differences in lipid metabolism in the course of tularemia in these two species. Hypertriglyceridemia and hypercholesterolemia developed in mice, while physiologically higher levels of triglycerides and cholesterol showed a decreasing tendency in common voles. On the other hand, the total plasma antioxidant capacity gradually dropped to 81.5% in mice on day 5 post-infection, while it increased to 130% on day 6 post-infection in common voles. Significant correlations between tissue bacterial burdens and several biochemical parameters were found. CONCLUSION: As differences in lipid metabolism and the total antioxidant capacity of highly susceptible rodent species were demonstrated, the role of triglycerides, cholesterol and antioxidants in tularemic sepsis should be further investigated.

• MeSH
• antioxidancia metabolismus   MeSH
• Arvicolinae mikrobiologie   MeSH
• cholesterol metabolismus   MeSH
• metabolismus lipidů MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• nemoci hlodavců metabolismus   mikrobiologie   MeSH
• triglyceridy metabolismus   MeSH
• tularemie metabolismus   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH
• srovnávací studie MeSH

• MeSH
• financování organizované MeSH
• Publikační typ
• abstrakty MeSH

Specific allergen immunotherapy is frequently associated with adverse reactions. Several strategies are being developed to reduce the allergenicity while maintaining the therapeutic benefits. Peptide immunotherapy is one such approach. Methods for the simple and rapid identification of immunogenic epitopes of allergens (i.e. allergenic epitopes) are ongoing and could potentially lead to peptide-based vaccines. An epitope extraction technique, based on biofunctionalized magnetic microspheres self-organized under a magnetic field in a channel of a simple microfluidic device fabricated from polydimethylsiloxane, was applied in the isolation and identification of prospective allergenic epitopes. Similarly to chromatographic column separations, the easily replaceable plug of self-organized beads in the channel benefits especially from an even larger surface-to-volume ratio and an enhanced interaction of the surfaces with passing samples. Ovalbumin, the major protein of egg white and a typical representative of food allergens, was selected as the model molecule. Highly resistant ovalbumin was at first efficiently digested by a magnetic proteolytic reactor with trypsin treated with l-1-tosylamido-2-phenylethyl chloromethyl ketone and the second step, i.e. capture of allergenic epitopes from the mixture of peptides, was performed by a magnetic immunoaffinity carrier with orientedly immobilized rabbit anti-ovalbumin IgG molecules. Captured peptides were released with 0.05% trifluoroacetic acid. The elution fractions were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The peptide fragment of ovalbumin HIATNAVLFFGR (m/z: 1345.75, position: 371-382) was identified as a relevant allergenic epitope in this way. Such a microfluidic magnetic force-based epitope extraction technique applied in the epitope mapping of ovalbumin has the potential to be a significant step towards developing safe and cost-effective epitope-based vaccines.

• MeSH
• alergeny chemie   imunologie   MeSH
• epitopy analýza   MeSH
• financování organizované MeSH
• hmotnostní spektrometrie MeSH
• imunomagnetická separace metody   MeSH
• mapování epitopu metody   MeSH
• mikrofluidní analytické techniky metody   MeSH
• mikrosféry MeSH
• ovalbumin chemie   imunologie   MeSH
• potravinová alergie MeSH
• vakcíny MeSH

Rapid and reliable detection, identification, and typing of bacterial species are necessary in response to natural or terrorist-caused outbreaks of infectious diseases and play crucial roles in diagnosis and efficient treatment. We report here two proteomic approaches with a high potential in the detection and identification of Coxiella burnetii, the causative agent of Q fever. The first of them starts with the acetonitrile (ACN) and trichloroacetic acid extractions of inactivated C. burnetii cells followed by the detection of extracted molecules and ions derived from the inactivated cells by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. In the second approach, identification of the proteins extracted by ACN is accomplished after enzymatic digestion by electrospray tandem mass spectrometry coupled to a nanoscale ultraperformance liquid chromatography (LC-MS/MS). In order to observe morphological differences on the surface structures upon extraction, the inactivated and treated cells of the bacterium were examined by electron microscopy. The LC-MS/MS approach has allowed identification of 20 proteins in the ACN extracts of C. burnetii strain RSA 493 that were observed in more than 3 out of 10 experiments.

• MeSH
• bakteriální proteiny MeSH
• chromatografie kapalinová MeSH
• Coxiella burnetii chemie   izolace a purifikace   MeSH
• databáze faktografické MeSH
• elektronová mikroskopie MeSH
• financování organizované MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• tandemová hmotnostní spektrometrie MeSH
• trypsin metabolismus   MeSH