Státní zdravotní ústav organizuje v rámci Systému monitorování zdravotního stavu obyvatelstva ČR ve vztahu k životnímu prostředí celostátní monitoring alergických onemocnění u dětí. Šetření probíhají od roku 1996 v sadě vybraných měst, ve které jsou zahrnuta také tři města ostravsko- karvinské aglomerace – Ostrava, Karviná a Frýdek-Místek, která patří dlouhodobě k oblastem s nejvíce znečištěným ovzduším v Evropě. Příspěvek obsahuje porovnání výskytu alergických onemocnění a astmatu v této polutanty zatížené oblasti s výskytem onemocnění v ostatních městech monitoringu. Zdrojem dat je výpis z dokumentace spolupracujících dětských lékařů a dotazník pro rodiče, který je vyplňován v rámci preventivních prohlídek v populaci 5, 9, 13 a 17letých dětí. Průměrná prevalence alergických onemocnění v ostravsko-karvinské aglomeraci 31,4 % dětí, zjištěná zatím posledním šetřením v roce 2006, se nelišila od prevalence v ostatních 15 městech monitoringu (31,9 %, p = 0,726). Signifikantní rozdíly však byly zjištěny ve výskytu některých diagnóz: astmatu (10,0 vs. 7,9 %; p < 0,014), alergické rýmy (20,3 % vs. 14,1 %; p < 0,001) a společného onemocnění dýchacích cest (astma s alergickou rýmou – 6,6 % vs. 3,1 %; p < 0,001). V souboru měst na Ostravsku bylo signifikantně více astmatických dětí s nedostatečnou kontrolou astmatu podle Testu kontroly astmatu oproti ostatním městům (40,5 % vs. 13,6 %; p < 0,001). Také výskyt respiračních příznaků, jako jsou pískoty mimo nachlazení a ponámahové pískoty (5,8 % vs. 3,3 %; p < 0,001 a 6,3 % vs. 3,6 %; p < 0,001) nebo přítomnost suchého nočního kašle (18,9 % vs. 10,5 %; p < 0,001) byly u dětí z měst ostravsko-karvinské aglomerace signifikantně vyšší.

National Institute of Public Health organizes nationwide monitoring of allergic diseases in children within the framework of the system of monitoring of health of population in the Czech Republic in relation to the environment. Monitoring has been in progress since 1996 in the set of selected cities, including three cities of the Ostrava, Karviná and Frýdek-Místek metro areas, belonging from a long-term view among areas with most polluted environment in Europe. This study compares the prevalence of allergic diseases and asthma in this pollution affected area with the prevalence of diseases in other cities involved in the monitoring project. Records taken from documentation of cooperating paediatricians and forms filled out by parents during preventive check-ups of children at the age of 5, 9, 13 and 17 years were used as data resources. The average prevalence of allergic diseases in the Ostrava-Karviná metro area of 31.4 % children as found in the last monitoring in 2006 did not differ from other 15 cities and towns included in the monitoring project (31.9 %, p = 0.726 ). Significant difference was however found in the prevalence of several diagnoses: asthma (10.0 vs. 7.9 %; p < 0.014), allergic rhinitis (20.3 % vs. 14.1 %; p < 0.001) and concurrent diseases of airways (asthma and allergic rhinitis – 6.6 % vs. 3.1 %; p < 0.001). According to the test of asthma control there were significantly more children with insufficient asthma control in cities and towns in Ostrava region compared to other cities and towns (40.5 % vs. 13.6 %). The prevalence of respiratory symptoms as wheezing without cold and post-stress wheezing (5.8 % vs. 3.3 %; p < 0.001 and 6.3 % vs. 3.6 %; p < 0.001) or the presence of night dry cough (18.9 % vs. 10.5 %; p < 0.001) was also significantly higher in the set of children in the Ostrava-Karviná region.

• Keywords
• astma,
• MeSH
• Hypersensitivity epidemiology   prevention & control   MeSH
• Asthma epidemiology   prevention & control   MeSH
• Child MeSH
• Epidemiologic Methods MeSH
• Humans MeSH
• Adolescent MeSH
• Environmental Monitoring methods   MeSH
• Immune System Diseases epidemiology   prevention & control   MeSH
• Pediatrics methods   MeSH
• Preventive Medicine MeSH
• Surveys and Questionnaires standards   utilization   MeSH
• Risk Factors MeSH
• Statistics as Topic MeSH
• Environmental Exposure prevention & control   MeSH
• Check Tag
• Child MeSH
• Humans MeSH
• Adolescent MeSH
• Geographicals
• Czech Republic MeSH

Cyklosporin A (CyA) je lék s úzkým terapeutickým rozmezím u něhož se projevuje významná interindividuální variabilita, která má za následek rozdílnou odpověď na terapii. V klinické praxi neexistují imunologické testy, kterými by bylo možno účinnost imunosuprese cyklosporinem sledovat, a proto se monitorování CyA omezuje na stanovení jeho koncentrací v krvi. Pro terapeutické monitorování (TDM) cyklosporinu A se používají především metody imunoanalytické, metody vysokoúčinné kapalinové chromatografie (HPLC) a kapalinová chromatografie ve spojení s hmotnostní detekcí (LC-MS, popř. LC-MS/MS). Výhodou imunoanalytických metod je analýza velkého množství vzorků během krátkého času a snadná ovladatelnost přístrojů. Jejich nevýhodou jsou zkřížené reakce s metabolity CyA. HPLC metody se považují za referenční metody stanovení CyA. Vyžadují však časově náročné extrakční postupy přípravy vzorků, které jsou důležité pro dobrou separaci. HPLC metody jsou také používány k měření CyA společně s jeho metabolity. V tom případě je celkový čas analýzy až několik desítek minut. Metody LC-MS a LC-MS/MS jsou selektivní, velmi citlivé a specifické. Výhodou těchto metod oproti klasické HPLC je zejména v případě LC-MS/MS rychlá průchodnost vzorků a možnost stanovení CyA s metabolity nebo s dalšími imunosupresivy v jedné analýze. Nevýhodou je vysoká pořizovací cena přístroje a nutnost odborné obsluhy.

Cyclosporine A (CyA) is a drug with narrow therapeutic range and high interindividual variability. No immunologic tests are known to verify immunosuppressive effect of cyclosporine in clinical practice and that's why CyA monitoring is limited mainly to determine its concentration in blood. Immunoanalytic methods, high performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS or LC-MS/MS) are regularly used to determine CyA concentration in therapeutic drug monitoring (TDM). Immunoanalytic methods can analyze a lot of samples in short time and they are easy practicability, but the disadvantage is crossed reactions with CyA metabolites. HPLC methods are considered to be referential for CyA determination, but they require time-consuming extraction procedures important for successful separation. The analysis may take even tens of minutes. HPLC methods are also used to determine CyA together with its metabolites. LC-MS and LC-MS/MS methods are selective, very sensitive and specific. The advantage of these methods especially in the case of LC-MS/MS is high sample throughput and the possibility to determine CyA simultaneously with its metabolites or other immunosuppressives in one analysis. On the other hand the initial instrumentation cost and a requirements of skilful staff are limitations of these methods.

• Keywords
• cyklosporin A, TDM,, imunoanalytické metody, HPLC-UV, LC-MS, LC-MS/MS,
• MeSH
• Cyclosporine isolation & purification   blood   MeSH
• Pharmacologic Actions MeSH
• Financing, Organized MeSH
• Mass Spectrometry methods   utilization   MeSH
• Immunoassay methods   utilization   MeSH
• Humans MeSH
• Drug Monitoring methods   utilization   MeSH
• Reference Standards MeSH
• Sirolimus analogs & derivatives   isolation & purification   blood   MeSH
• Tacrolimus isolation & purification   blood   MeSH
• Chromatography, High Pressure Liquid methods   utilization   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Review MeSH

There is an increasing need for proper quality control tools in the pre-analytical phase of the molecular diagnostic workflow. The aim of the present study was to identify biomarkers for monitoring pre-analytical mRNA quality variations in two different types of blood collection tubes, K2EDTA (EDTA) tubes and PAXgene Blood RNA Tubes (PAXgene tubes). These tubes are extensively used both in the diagnostic setting as well as for research biobank samples. Blood specimens collected in the two different blood collection tubes were stored for varying times at different temperatures, and microarray analysis was performed on resultant extracted RNA. A large set of potential mRNA quality biomarkers for monitoring post-phlebotomy gene expression changes and mRNA degradation in blood was identified. qPCR assays for the potential biomarkers and a set of relevant reference genes were generated and used to pre-validate a sub-set of the selected biomarkers. The assay precision of the potential qPCR based biomarkers was determined, and a final validation of the selected quality biomarkers using the developed qPCR assays and blood samples from 60 healthy additional subjects was performed. In total, four mRNA quality biomarkers (USP32, LMNA, FOSB, TNRFSF10C) were successfully validated. We suggest here the use of these blood mRNA quality biomarkers for validating an experimental pre-analytical workflow. These biomarkers were further evaluated in the 2nd ring trial of the SPIDIA-RNA Program which demonstrated that these biomarkers can be used as quality control tools for mRNA analyses from blood samples.

• MeSH
• Humans MeSH
• RNA, Messenger blood   chemistry   genetics   isolation & purification   MeSH
• Blood Specimen Collection MeSH
• Polymerase Chain Reaction MeSH
• Oligonucleotide Array Sequence Analysis MeSH
• RNA Stability MeSH
• Gene Expression Profiling MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Validation Study MeSH

Úvod: Tumorigeneze je spojena s deregulací Wnt signalizační dráhy, která se významně podílí na regulaci proliferace a diferenciace buněk. Mutace nebo deregulace exprese komponent Wnt dráhy se uplatňují v indukci řady chorob včetně vzniku karcinomu. V případě vzniku neoplastických změn ve střevu dochází k aberantní stabilizaci a akumulaci ß-kateninu, který i v nepřítomnosti Wnt signálů uniká destrukci a vstupuje do buněčného jádra. Zde vytváří transkripčně aktivní komplex, a tak dochází k zahájení transkripce řady genů včetně protoonkogenů. Otázkou je, co způsobuje akumulaci ß-kateninu (mutace nebo deregulace exprese určitých komponent Wnt signalizace) a zda deregulace v kanonické Wnt/ß-kateninové signalizaci není spojena se změnami nekanonické Wnt signalizace, která by se rovněž mohla uplatňovat v tumorigenezi. Cíl studie: Cílem práce bylo porovnat expresi vybraných komponent Wnt signalizace ve vzorcích neoplastické tkáně pacientů s kolorektálním karcinomem a v okolní makroskopicky zdravé tkáni. Materiál a metodika: Hladiny transkriptů vybraných genů Wnt dráhy byly měřeny pomocí kvantitativní PCR ve vzorcích tkáně získané od pacientů operovaných pro kolorektální karcinom. Výsledky: Neoplastická tkáň vykazovala zvýšenou hladinu transkriptů genů pro Axin 2, DKK1 a NKD1 a sníženou hladinu transkriptu pro sFRP1. U dalších studovaných genů (Axin 1, LRP5, ßTrCP, DKK2, DKK3 a ß-katenin) byly hladiny transkriptů v neoplastické i v okolní zdravé tkáni stejné. Závěr: Prokázali jsme, že neoplastická tkáň pacientů s kolorektálním karcinomem, která je běžně spojena s aktivací kanonické Wnt/ß-kateninové dráhy, vykazuje na úrovni mRNA změny v expresi 4 komponent Wnt signalizace, které působí tlumivě na Wnt/ß-kateninovou dráhu. Nález zvýšené hladiny NKD1 mRNA spolu s poklesem hladiny sFRP1 naznačuje posun Wnt odpovědí ve směru k nekanonické dráze, což by se mohlo uplatňovat v tumorigenezi. Data rovněž naznačují možnost použití NKD1 mRNA a sFRP1 mRNA jako markerů kolorektální karcinogeneze.

Introduction: The tumorigenesis is associated with deregulation of Wnt signaling pathway which is mostly involved in regulation of cell proliferation and differentiation. Mutation or deregulations of expression of the Wnt pathway components are able to inducel diseases including development of carcinoma. In case of the development of neoplastic changes in colon ß-catenin is stabilized and accumulated in cytosol without Wnt signaling due to a decreased rate of its degradation. It is translocated to the cell nucleus where it forms a transcriptionally complex and initiates inappropriate transcription of several genes including protooncogenes. The question is, what is the cause of ß-catenin accumulation (mutation or deregulation of expression of certain components of Wnt pathway) and whether the deregulation of canonic Wnt/ß-catenin signaling is accompanied with changes of noncanonical Wnt signaling pathways could also be involved in tumorigenesis. The aim of the study: The aim of the study was to compare the expression of selected components of Wnt signaling in specimens of neoplastic tissue and surrounding healthy tissue of patients with colorectal cancer. Material and methods: Transcript levels of selected genes of Wnt pathway were measured by quantitative PCT in specimens of tissue from patients operated for colorectal cancer. Results: Neoplastic tissue showed increased up-regulation of transcript genes for Axin 2, DKK1 a NKD1 and down-regulation for sFRP1 transcript. Other selected genes (Axin 1, LRP5, ßTrCP, DKK2, DKK3 a ß-katenin) showed no significant changes between neoplastic and normal tissue. Conclusion: We demonstrated that neoplastic tissue of patients with colorectal cancer, that is commonly associated with canonical Wnt/ß-catenin pathway, had increased mRNA expression of 4 components of Wnt signaling which are known to cause inactivation of Wnt/ß-catenin pathway. The finding of NKD1 mRNA up-regulation and simultaneous sFRP1 mRNA down-regulation suggests the shift of Wnt response to the noncanonical pathway which might play a role in tumorigenesis. Our data suggest the possibility of using NKD1 mRNA and sFRP1 mRNA as markers of colorectal carcinogenesis.

• MeSH
• Gene Expression genetics   immunology   MeSH
• Financing, Organized MeSH
• Colorectal Neoplasms etiology   genetics   MeSH
• Humans MeSH
• Specimen Handling methods   utilization   MeSH
• Reverse Transcriptase Polymerase Chain Reaction methods   instrumentation   utilization   MeSH
• Wnt Proteins genetics   immunology   MeSH
• Gene Expression Profiling methods   utilization   MeSH
• Statistics as Topic MeSH
• Check Tag
• Humans MeSH

• MeSH
• Anesthesia, General methods   nursing   MeSH
• Child MeSH
• Risk Assessment MeSH
• Clinical Laboratory Techniques utilization   MeSH
• Drug Monitoring methods   utilization   MeSH
• Pediatrics manpower   MeSH
• Preoperative Care methods   standards   MeSH
• Adverse Drug Reaction Reporting Systems organization & administration   utilization   MeSH
• Patient Selection MeSH
• Check Tag
• Child MeSH
• Publication type
• Practice Guideline MeSH

Human multipotent neural stem cells could effectively be used for the treatment of a variety of neurological disorders. However, a defining signature of neural stem cell lines that would be expandable, non-tumorigenic, and differentiate into desirable neuronal/glial phenotype after in vivo grafting is not yet defined. Employing a mass spectrometry approach, based on selected reaction monitoring, we tested a panel of well-described culture conditions, and measured levels of protein markers routinely used to probe neural differentiation, i.e. POU5F1 (OCT4), SOX2, NES, DCX, TUBB3, MAP2, S100B, GFAP, GALC, and OLIG1. Our multiplexed assay enabled us to simultaneously identify the presence of pluripotent, multipotent, and lineage-committed neural cells, thus representing a powerful tool to optimize novel and highly specific propagation and differentiation protocols. The multiplexing capacity of this method permits the addition of other newly identified cell type-specific markers to further increase the specificity and quantitative accuracy in detecting targeted cell populations. Such an expandable assay may gain the advantage over traditional antibody-based assays, and represents a method of choice for quality control of neural stem cell lines intended for clinical use.

• MeSH
• Biomarkers MeSH
• Cell Differentiation * MeSH
• Cell Line MeSH
• Cell Lineage genetics   MeSH
• Mass Spectrometry MeSH
• Immunohistochemistry MeSH
• Humans MeSH
• Neural Stem Cells cytology   metabolism   MeSH
• Neuroglia MeSH
• Neurons MeSH
• Gene Expression Profiling MeSH
• Gene Expression Regulation, Developmental MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Amphibians are globally threatened, but not all species are affected equally by different threatening processes. This is true for the threat posed by the chytridiomycete fungus (Batrachochytrium dendrobatidis). We compiled a European data set for B. dendrobatidis to analyze the trends of infection in European amphibians. The risk of infection was not randomly distributed geographically or taxonomically across Europe. Within countries with different prevalence, infection was nonrandom in certain amphibian taxa. Brown frogs of the genus Rana were unlikely to be infected, whereas frogs in the families Alytidae and Bombinatoridae were significantly more likely to be infected than predicted by chance. Frogs in the 2 families susceptible to B. dendrobatidis should form the core of attempts to develop spatial surveillance studies of chytridiomycosis in Europe. Ideally, surveys for B. dendrobatidis should be augmented by sampling the widespread genus Pelophylax because this taxon exhibits geographically inconsistent overinfection with B. dendrobatidis and surveillance of it may facilitate recognition of factors causing spatial variability of infection intensity. Several European amphibian taxa were not represented in our data set; however, surveillance of unsampled species should also occur when warranted.

• MeSH
• Chytridiomycota isolation & purification   physiology   MeSH
• Risk Assessment MeSH
• Mycoses epidemiology   microbiology   MeSH
• Amphibians microbiology   MeSH
• Polymerase Chain Reaction veterinary   MeSH
• Prevalence MeSH
• Conservation of Natural Resources * MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Europe MeSH

Flow injection analysis (FIA) is an analytical method where the reaction mixture is injected into flow of liquid. The reaction product is monitored by a suitable detector such as ultraviolet/visible (UV/VIS) spectrophotometric or electrochemical detector. Mass spectrometric detectors (MS) are coming to be a standard equipment of analytical laboratories in the present time. This work is focused on application of FIA-MS instrumentation for monitoring of Ellman's reaction where both reactants (acetylthiocholine and 5,5’-dithiobis-2-nitrobenzoic acid, DTNB) and the reaction product (5-mercapto-2-nitrobenzoic acid) are monitored. This reaction is usually used for monitoring of acetylcholinesterase and butyrylcholinesterase. Due to its simplicity, the developed method is generally applicable for monitoring of enzymatic reactions of cholinesterases. The main advantage of this method is high selectivity and reduction of influence of compounds, which are reacting with DTNB, resulting in a color product of Ellman's reaction.

• MeSH
• Acetylcholine chemistry   metabolism   MeSH
• Acetylcholinesterase * analysis   metabolism   MeSH
• Time Factors MeSH
• Cholinesterase Inhibitors chemistry   metabolism   MeSH
• Mass Spectrometry * methods   MeSH
• Hydrolysis MeSH
• Flow Injection Analysis * methods   MeSH
• Spectrophotometry methods   MeSH

Many severe diseases of the respiratory tract lead to hospitalisation. These diseases are often caused by viral infections and may cause increased mortality. The most common viral pathogens involved in these cases, which are also associated with significant morbidity and mortality during the influenza seasons are influenza viruses. Rapid differential diagnosis of influenza viruses is therefore of great importance. Classical diagnosis of these viruses involves virus cultures. Of the rapid diagnostic methodologies which have been developed are RT-PCR, multiplex PCR, real-time PCR. In the present study we have monitored clinical samples from patients of different age groups from selected regions in Slovakia and compared the effectiveness of the classical and molecular biological diagnostic methods. The molecular biological methods proved to be rapid, accurate and effective. Application of these techniques in diagnosis of the respiratory illnesses should help in the prevention, therapy and disease control.

• MeSH
• Acute Disease MeSH
• Influenza, Human virology   MeSH
• Diagnosis, Differential MeSH
• Respiratory Tract Infections virology   MeSH
• Virus Cultivation MeSH
• Humans MeSH
• Polymerase Chain Reaction methods   MeSH
• RNA, Viral analysis   MeSH
• Seasons MeSH
• Check Tag
• Humans MeSH

• MeSH
• Environmental Monitoring MeSH
• Environmental Exposure MeSH
• Environmental Pollution MeSH
• Geographicals
• Czech Republic MeSH

• Conspectus
• Veřejné zdraví a hygiena
• NML Fields
• environmentální vědy