Targeted proteomics recently proved to be a technique for the detection and absolute quantification of proteins not easily accessible to classical bottom-up approaches. Due to this, it has been considered as a high fidelity tool to detect potential warfare agents in wide spread kinds of biological and environmental matrices. Clostridium perfringens toxins are considered to be potential biological weapons, especially the epsilon toxin which belongs to a group of the most powerful bacterial toxins. Here, the development of a target mass spectrometry method for the detection of C. perfringens protein toxins (alpha, beta, beta2, epsilon, iota) is described. A high-resolution mass spectrometer with a quadrupole-Orbitrap system operating in target acquisition mode (parallel reaction monitoring) was utilized. Because of the lack of commercial protein toxin standards recombinant toxins were prepared within Escherichia coli. The analysis was performed using proteotypic peptides as the target compounds together with their isotopically labeled synthetic analogues as internal standards. Calibration curves were calculated for each peptide in concentrations ranging from 0.635 to 1101 fmol/μL. Limits of detection and quantification were determined for each peptide in blank matrices.

• MeSH
• bakteriální proteiny analýza   genetika   MeSH
• bakteriální toxiny analýza   genetika   MeSH
• chromatografie kapalinová MeSH
• Clostridium perfringens * genetika   růst a vývoj   metabolismus   MeSH
• Escherichia coli genetika   MeSH
• peptidy analýza   genetika   MeSH
• proteomika MeSH
• rekombinantní proteiny analýza   MeSH
• tandemová hmotnostní spektrometrie MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

We have studied the biodiversity of trypanosomes from birds and bloodsucking Diptera on a large number of isolates. We used two molecular approaches, random amplification of polymorphic DNA (RAPD) method, and sequence analysis of the small subunit ribosomal RNA (SSU rRNA) gene. RAPD method divided the isolates into 11 separate lineages. Phylogenetic analysis of the SSU rRNA gene was congruent with the RAPD. Morphometric analysis of kinetoplast width and cell length was in agreement with molecular data. Avian trypanosomes appeared polyphyletic on SSU rDNA tree; thus, they do not represent a taxonomic group. We propose that all lineages recovered by SSU analysis probably represent distinct species of avian trypanosomes. We discuss possible transmission ways and geographical distribution of new avian trypanosome lineages. Finally, we recommend methods that should be used for species determination of avian trypanosomes.

• MeSH
• biodiverzita MeSH
• DNA primery MeSH
• fylogeneze MeSH
• protozoální DNA genetika   MeSH
• ptáci parazitologie   MeSH
• RNA ribozomální genetika   MeSH
• sekvenční analýza DNA MeSH
• technika náhodné amplifikace polymorfní DNA metody   MeSH
• transmisní elektronová mikroskopie MeSH
• Trypanosoma klasifikace   genetika   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Metabolomics has become an important tool in clinical research and diagnosis of human diseases. In this work we focused on the diagnosis of inherited metabolic disorders (IMDs) in plasma samples using a targeted metabolomic approach. The plasma samples were analyzed with the flow injection analysis method. All the experiments were performed on a QTRAP 5500 tandem mass spectrometer (AB SCIEX, U.S.A.) with electrospray ionization. The compounds were measured in a multiple reaction monitoring mode. We analyzed 50 control samples and 34 samples with defects in amino acid metabolism (phenylketonuria, maple syrup urine disease, tyrosinemia I, argininemia, homocystinuria, carbamoyl phosphate synthetase deficiency, ornithine transcarbamylase deficiency, nonketotic hyperglycinemia), organic acidurias (methylmalonic aciduria, propionic aciduria, glutaric aciduria I, 3-hydroxy-3-methylglutaric aciduria, isovaleric aciduria), and mitochondrial defects (medium-chain acyl-coenzyme A dehydrogenase deficiency, carnitine palmitoyltransferase II deficiency). The controls were distinguished from the patient samples by principal component analysis and hierarchical clustering. Approximately 80% of patients were clearly detected by absolute metabolite concentrations, the sum of variance for first two principle components was in the range of 44-55%. Other patient samples were assigned due to the characteristic ratio of metabolites (the sum of variance for first two principle components 77 and 83%). This study has revealed that targeted metabolomic tools with automated and unsupervised processing can be applied for the diagnosis of various IMDs.

• MeSH
• analýza hlavních komponent MeSH
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• metabolom MeSH
• metabolomika metody   MeSH
• mladiství MeSH
• předškolní dítě MeSH
• průtoková injekční analýza MeSH
• reprodukovatelnost výsledků MeSH
• shluková analýza MeSH
• tandemová hmotnostní spektrometrie MeSH
• vrozené poruchy metabolismu aminokyselin krev   diagnóza   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

A novel avian trypanosome, Trypanosoma culicavium sp. nov., isolated from Culex mosquitoes, is described on the basis of naturally and experimentally infected vectors and bird hosts, localization in the vector, morphological characters and molecular data. This study provides the first comprehensive description of a trypanosome species transmitted by mosquitoes, in which parasites form plugs and rosettes on the stomodeal valve. Trypanosomes occurred as long epimastigotes and short trypomastigotes in vectors and culture and as long trypomastigotes in birds. Transmission of parasites to bird hosts was achieved exclusively by ingestion of experimentally infected Culex mosquito females by canaries (Serinus canaria), but not by Japanese quails (Coturnix japonica), nor by the bite of infected vectors, nor by ingestion of parasites from laboratory cultures. Transmission experiments and the identity of isolates from collared flycatchers (Ficedula albicollis) and Culex mosquitoes suggests that the natural hosts of T. culicavium are insectivorous songbirds (Passeriformes). Phylogenetic analyses of small-subunit rRNA and glycosomal glyceraldehyde-3-phosphate dehydrogenase gene sequences demonstrated that T. culicavium sp. nov. is more related to Trypanosoma corvi than to other avian trypanosomes (e.g. Trypanosoma avium and Trypanosoma bennetti).

• MeSH
• Culex parazitologie   MeSH
• fylogeneze MeSH
• geny rRNA MeSH
• mezerníky ribozomální DNA chemie   genetika   MeSH
• mikroskopie MeSH
• molekulární sekvence - údaje MeSH
• protozoální DNA chemie   genetika   MeSH
• ptáci parazitologie   MeSH
• ribozomální DNA chemie   genetika   MeSH
• RNA protozoální genetika   MeSH
• RNA ribozomální genetika   MeSH
• sekvenční analýza DNA MeSH
• shluková analýza MeSH
• Trypanosoma klasifikace   genetika   izolace a purifikace   patogenita   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Phlebotomine sand flies are blood-sucking insects transmitting Leishmania parasites. In bitten hosts, sand fly saliva elicits specific immune response and the humoral immunity was shown to reflect the intensity of sand fly exposure. Thus, anti-saliva antibodies were suggested as the potential risk marker of Leishmania transmission. In this study, we examined the long-term kinetics and persistence of anti-Phlebotomus papatasi saliva antibody response in BALB/c and C57BL/6 mice. We also tested the reactivity of mice sera with P. papatasi salivary antigens and with the recombinant proteins. METHODOLOGY/PRINCIPAL FINDINGS: Sera of BALB/c and C57BL/6 mice experimentally bitten by Phlebotomus papatasi were tested by ELISA for the presence of anti-saliva IgE, IgG and its subclasses. We detected a significant increase of specific IgG and IgG1 in both mice strains and IgG2b in BALB/c mice that positively correlated with the number of blood-fed P. papatasi females. Using western blot and mass spectrometry we identified the major P. papatasi antigens as Yellow-related proteins, D7-related proteins, antigen 5-related proteins and SP-15-like proteins. We therefore tested the reactivity of mice sera with four P. papatasi recombinant proteins coding for most of these potential antigens (PpSP44, PpSP42, PpSP30, and PpSP28). Each mouse serum reacted with at least one of the recombinant protein tested, although none of the recombinant proteins were recognized by all sera. CONCLUSIONS: Our data confirmed the concept of using anti-sand fly saliva antibodies as a marker of sand fly exposure in Phlebotomus papatasi-mice model. As screening of specific antibodies is limited by the availability of salivary gland homogenate, utilization of recombinant proteins in such studies would be beneficial. Our present work demonstrates the feasibility of this implementation. A combination of recombinant salivary proteins is recommended for evaluation of intensity of sand fly exposure in endemic areas and for estimation of risk of Leishmania transmission.

• MeSH
• časové faktory MeSH
• ELISA MeSH
• hmyzí proteiny imunologie   MeSH
• imunoglobulin E krev   MeSH
• imunoglobulin G krev   MeSH
• myši inbrední BALB C MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• Phlebotomus imunologie   MeSH
• slinné proteiny a peptidy imunologie   MeSH
• tvorba protilátek MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Monoxenous trypanosomatid Herpetomonas trimorpha sp. nov. was isolated from the digestive tract of the biting midge Culicoides truncorum (Ceratopogonidae, Diptera). This species forms three distinct morphotypes in culture: the microflagellate promastigote, the small promastigote and the long promastigote. The last form is unique for the newly described species. Phylogenetic analyses of SSU rRNA and glycosomal glyceraldehyde phosphate dehydrogenase genes showed that H. trimorpha sp. nov. is the closest relative of Herpetomonas ztiplika, another monoxenous trypanosomatid isolated from biting midges. However, morphological and randomly amplified polymorphic DNA analyses confirmed that H. trimorpha sp. nov. is distinct from H. ztiplika.

• MeSH
• Ceratopogonidae parazitologie   MeSH
• fylogeneze MeSH
• gastrointestinální trakt parazitologie   MeSH
• glyceraldehyd-3-fosfátdehydrogenasy genetika   MeSH
• molekulární sekvence - údaje MeSH
• protozoální DNA genetika   MeSH
• protozoální proteiny genetika   MeSH
• ribozomální DNA genetika   MeSH
• Trypanosomatina klasifikace   enzymologie   genetika   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• Publikační typ
• abstrakty MeSH

Cíl studie: Přehled současných poznatků o dědičných metabolických poruchách u těhotných a u plodů. Typ studie: Literární přehled. Název a sídlo pracoviště: Laboratoř dědičných metabolických poruch, Oddělení klinické biochemie a Dětská klinika, Fakultní nemocnice a Univerzita Palackého, Olomouc. Metodika: Rešerše studií a publikací za pomoci medicínských databází. Závěr: K metabolickému vyšetření matky nebo dítěte je vhodné přistoupit, kdykoli se vyskytnou klinické problémy během nebo těsně po těhotenství. Většinu dědičných metabolických poruch dnes lze prenatálně diagnostikovat v prvním nebo druhém trimestru.

Objective: Review of contemporary knowledge about inherited metabolic disorders in pregnant women and fetuses. Design: A review of literature. Setting: Laboratory for inherited metabolic disorders, Departament of Clinical Biochemistry and Childern's Clinic, University Hospital and Palacky University, Olomouc. Method: Review of literature and publications using medical databases. Conclusion: Metabolic examination of mother or child is recommended if any clinical problems during or immediately after pregnancy occur. Nowadays most of inherited errors of metabolism can be diagnosed prenataly in first or second trimester of pregnancy.

• MeSH
• fenylketonurie v těhotenství MeSH
• financování organizované MeSH
• galaktosemie komplikace   MeSH
• glykogenóza typu I komplikace   MeSH
• HELLP syndrom diagnóza   MeSH
• infertilita etiologie   genetika   MeSH
• komplikace těhotenství genetika   klasifikace   MeSH
• lidé MeSH
• prenatální diagnóza využití   MeSH
• těhotenství MeSH
• vrozené poruchy metabolismu genetika   komplikace   MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• přehledy MeSH

Ke studiu ateromatózních změn v tepenné stěně, stanovení doby vhodné pro angioplastiku, bude použita rotační angiografie s trojrozměrným zobrazením cévní stěny rentgenovou technikou programu CalciView. Trojrozměrné zobrazování tepenného zúžení systémem Integris 3D-RA lze využít k detekci ateromatózních plátů v cévní stěně pomocí měkké techniky programu CalciView. K studii ateromatózních změn arteriální stěny, detekci sklerotických kalcifikovaných plátů v karotidách bude použito nativní RTG zobrazení s 3D rekonstrukcí, parametry budou srovnány s metodou IVUS.; A new soft X - ray technique with Calci-view programme will be used in detection and quantification of atheroclerotic lesions in vessel wall Three dimensional imaging of vessel narrowing with Calci-view programme can be used in detection of atherosclerotic plaque in blood vessel wall IVUS, CTA and AG parameters of sclerotic plaque will be compared with those of native 3DRA with Calci-view programme

• MeSH
• angiografie metody   MeSH
• angioplastika MeSH
• arteriální okluzní nemoci diagnóza   MeSH
• radiografie metody   MeSH
• skleroterapie MeSH
• ultrasonografie metody   MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• angiologie
• radiologie, nukleární medicína a zobrazovací metody
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

Three strains of a trypanosomatid protozoan were isolated from the midguts of two naturally infected species of biting midges [Culicoides (Oecacta) festivipennis and Culicoides (Oecacta) truncorum] and characterized by light and electron microscopy and by molecular techniques. Morphological characteristics and sequences of the 18S rRNA, 5S rRNA, spliced leader RNA and glycosomal glyceraldehyde-3-phosphate dehydrogenase genes indicate that the studied flagellates represent a novel phylogenetic lineage within the Trypanosomatidae. Based on phylogenetic analyses, the novel endosymbiont-free, monoxenous trypanosomatid was classified as Sergeia podlipaevi gen. nov., sp. nov. Interestingly, it is closely related to another trypanosomatid species that parasitizes the sand fly Lutzomyia evansi, a blood-sucking dipteran from South America. The type strain of S. podlipaevi sp. nov., ICUL/CZ/2000/CER3, was obtained from Malpighian tubes. Of 2518 females of seven species of biting midges trapped in the Czech Republic, more than 1.5 % were infected by trypanosomatid parasites. An unrelated insect species, Culicoides (Monoculicoides) nubeculosus, was experimentally infected with S. podlipaevi, demonstrating that its host range extends to different subgenera of biting midges.

• MeSH
• Ceratopogonidae cytologie   parazitologie   ultrastruktura   MeSH
• elektronová mikroskopie MeSH
• financování organizované MeSH
• fylogeneze MeSH
• gastrointestinální trakt parazitologie   MeSH
• geny rRNA MeSH
• glyceraldehyd-3-fosfátdehydrogenasa (fosforylační) genetika   MeSH
• kinetoplastová DNA analýza   MeSH
• molekulární sekvence - údaje MeSH
• protozoální DNA genetika   chemie   MeSH
• ribozomální DNA genetika   chemie   MeSH
• RNA protozoální genetika   MeSH
• RNA ribozomální 18S genetika   MeSH
• RNA ribozomální 5S genetika   MeSH
• sekvenční analýza DNA MeSH
• sekvenční homologie nukleových kyselin MeSH
• Trypanosomatina cytologie   genetika   izolace a purifikace   klasifikace   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH