In order to evaluate the applicability of the cytokinesis-block micronucleus cytome assay in routine mutagenicity testing we investigated with this method different chemicals having different mechanisms of action: non-mutagens, direct-acting basealtering mutagens, direct-acting cross-linking mutagens, clastogens including a radiomimetic chemical, indirect-acting spindle poisons and indirect-acting enzyme inhibitors. We looked at the presence of micronuclei as biomarkers for either the loss of chromosome fragments (clastogen) or the loss of a whole chromosome (aneugen), nucleoplasmic bridges as biomarkers for complex rearrangements (e.g., dicentric chromosomes) and nuclear buds as biomarkers for gene amplification. The cytome assay proved to be a suitable tool to investigate genetic effects of environmental agents and to provide insight into their working mechanisms as all chemicals tested showed the expected response.

• MeSH
• buněčné linie MeSH
• cytokineze účinky léků   MeSH
• lidé MeSH
• mikrojaderné testy metody   MeSH
• mikrojádra chromozomálně defektní účinky léků   MeSH
• mutageny toxicita   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

PURPOSE: To assess whether the micronucleus cytome assay (MCyt) reliably detects DNA damage occurring in control and pathological superficial epithelial cells from human conjunctiva. METHODS: Impression cytology samples from the bulbar conjunctiva of 33 healthy controls, eight patients with conjunctival intraepithelial neoplasia (CIN) and eight with mucous membrane pemphigoid (MMP) were examined using the MCyt modified for the ocular surface. RESULTS: The mean number of micronuclei (MNi) in control samples was 0.94 MNi/1000 epithelial cells, with no significant difference between conjunctival quadrants and independent of sex and age. The MCyt assay applied to CIN-affected eyes showed a significantly higher frequency of MNi (18.63/1000 cells), apoptotic cells, nuclear enlargement, multinucleated cells, and keratolysis compared with the corresponding unaffected paired eyes and with the control value. Although the mean MNi frequency in MMP eyes was also higher (1.73 MNi/1000 cells), it did not prove to be statistically different from the control samples. On the other hand, the MMP-affected eyes revealed significantly elevated percentages of cells with snake-like chromatin, multinucleated cells, apoptotic cells, and nuclear buds compared with controls. CONCLUSIONS: Micronucleus cytome assay was adapted as a rapid screening test for genomic instability on the ocular surface. We have determined reference levels for MNi and other nuclear alterations on healthy conjunctiva and demonstrated that particularly frequencies of MNi are significantly elevated in conjunctiva affected by CIN. We demonstrate that MNi are more specific than other nuclear abnormalities and thus can be used for screening of ocular surface neoplasia.

• MeSH
• buněčné jádro MeSH
• epitelové buňky * MeSH
• konjunktiva MeSH
• lidé MeSH
• mikrojaderné testy MeSH
• poškození DNA * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• cytokineze genetika   účinky záření   MeSH
• financování organizované MeSH
• ionizující záření MeSH
• krev účinky záření   MeSH
• lidé MeSH
• lymfocyty cytologie   účinky záření   MeSH
• mikrojaderné testy metody   využití   MeSH
• radiační účinky MeSH
• radiometrie metody   využití   MeSH
• statistika jako téma MeSH
• struktury buněčného jádra genetika   účinky záření   MeSH
• Check Tag
• lidé MeSH

Plant-derived smoke and certain smoke compounds improve seed germination and enhance seedling growth of many species. Thus, smoke-infused water and the active smoke-derived compounds have the potential to be used in different agricultural and horticultural applications. However, despite these interesting and potentially practical properties, it should also be ascertained whether such compounds may pose a health risk, particularly if they are to be used in the production of food or fodder crops. Amongst some of the aspects that would be important to understand are any possible genotoxic properties that the compounds may possess due to potential carry-over effects. Here, we report on a genotoxicity study of 3,4,5-trimethylfuran-2(5H)-one, a compound from plant-derived smoke previously shown to have germination inhibitory activity. Using two in vitro tests, namely the bacterial VITOTOX® test (with/without S9 metabolic activation) and the cytome assay on human C3A cells, no genotoxicity or toxicity was found. Furthermore, these results support a previous study where a related smoke-derived compound with germination promoting properties was investigated.

• MeSH
• 4-nitrochinolin-1-oxid toxicita   MeSH
• benzopyreny toxicita   MeSH
• chinolony toxicita   MeSH
• furany farmakologie   MeSH
• hepatocyty cytologie   účinky léků   metabolismus   MeSH
• klíčení účinky léků   MeSH
• lidé MeSH
• luciferasy genetika   metabolismus   MeSH
• mutageny farmakologie   MeSH
• nádorové buněčné linie MeSH
• reportérové geny MeSH
• rostliny účinky léků   MeSH
• Salmonella typhimurium účinky léků   genetika   růst a vývoj   MeSH
• semena rostlinná účinky léků   růst a vývoj   MeSH
• SOS odpověď (genetika) účinky léků   MeSH
• testy genotoxicity MeSH
• transkripční faktory genetika   metabolismus   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In the last decade, it has become evident that complex mixtures of cyanobacterial bioactive substances, simultaneously present in blooms, often exert adverse effects that are different from those of pure cyanotoxins, and awareness has been raised on the importance of studying complex mixtures and chemical interactions. We aimed to investigate cytotoxic and genotoxic effects of complex extracts from laboratory cultures of cyanobacterial species from different orders (Cylindrospermopsis raciborskii, Aphanizomenon gracile, Microcystis aeruginosa, M. viridis, M. ichtyoblabe, Planktothrix agardhii, Limnothrix redekei) and algae (Desmodesmus quadricauda), and examine possible relationships between the observed effects and toxin and retinoic acid (RA) content in the extracts. The cytotoxic and genotoxic effects of the extracts were studied in the human hepatocellular carcinoma HepG2 cell line, using the MTT assay, and the comet and cytokinesis-block micronucleus (cytome) assays, respectively. Liquid chromatography electrospray ionization mass spectrometry (LC/ESI-MS) was used to detect toxins (microcystins (MC-LR, MC-RR, MC-YR) and cylindrospermopsin) and RAs (ATRA and 9cis-RA) in the extracts. Six out of eight extracts were cytotoxic (0.04-2 mgDM/mL), and five induced DNA strand breaks at non-cytotoxic concentrations (0.2-2 mgDM/mL). The extracts with genotoxic activity also had the highest content of RAs and there was a linear association between RA content and genotoxicity, indicating their possible involvement; however further research is needed to identify and confirm the compounds involved and to elucidate possible genotoxic effects of RAs.

• MeSH
• alkaloidy izolace a purifikace   toxicita   MeSH
• buňky Hep G2 MeSH
• Chlorophyta metabolismus   MeSH
• chromatografie kapalinová MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
• kometový test MeSH
• lidé MeSH
• mikrocystiny izolace a purifikace   toxicita   MeSH
• mikrojaderné testy MeSH
• mikrojádra chromozomálně defektní chemicky indukované   MeSH
• poškození DNA * MeSH
• sinice metabolismus   MeSH
• tandemová hmotnostní spektrometrie MeSH
• tretinoin izolace a purifikace   toxicita   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The lymphocyte cytokinesis-block micronucleus (CBMN) assay has been applied in hundreds of in vivo biomonitoring studies of humans exposed either environmentally or occupationally to genotoxic chemicals. However, there is an emerging need to re-evaluate the use of MN and other biomarkers within the lymphocyte CBMN cytome assay as quantitative indicators of exposure to main classes of chemical genotoxins. The main aim of the present report is to systematically review published studies investigating the use of the lymphocyte CBMN assay to determine DNA damage in subjects exposed to anaesthetic gases. We also compared performance of the CBMN assay with other DNA damage assays employed and identified strengths and weaknesses of the published studies. We have retrieved 11 studies, published between 1996 and 2013, reporting MN associated with occupational exposures (operating room personnel). The individual job categories were often described (anaesthesiologists, technicians, radiologists) among cases, as well as duration of exposure. All studies reported the compounds present at the workplace and, in some instances, the exposure levels were measured. Controls were usually recruited among personnel at the hospital not exposed to anaesthetics or they were healthy unexposed subjects from general population. The number of investigated subjects, due to the character of the occupation, was relatively smaller than those investigated in other occupational monitoring settings. Overall, the majority of the studies were age- and gender- matched (or investigated only males or females) while less attention was given to lifestyle confounders. Appropriate measurement of exposure, available in approximately half of the studies only, was compromised by the lack of the personal dosimetry-based determinations. In all studies, higher MN frequencies were observed in exposed individuals. The meta-analysis of mean MN frequency of combined studies confirmed this tendency (log mean ratio=0.56 [0.34-0.77]; P=3.51×10(-7)). Similar differences between the exposed and controls were also observed for other biomarkers.

• MeSH
• anestetika inhalační škodlivé účinky   MeSH
• chromozomální aberace * MeSH
• cytokineze * MeSH
• lidé MeSH
• lymfocyty účinky léků   ultrastruktura   MeSH
• pracovní expozice * MeSH
• zdravotnický personál * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Monoklonální gamapatie nejasného významu (MGUS) je prekancerózou, která se nejčastěji vyvíjí do mnohočetného myelomu (MM), druhého nejčastějšího krevního nádorového onemocnění. Projekt MGUS 2010 je prvním projektem České myelomové skupiny, respektive myelomové sekce České hematologické společnosti, čerpající z Registru monoklonáních gamapatií vzniklého v roce 2007. Hlavní cíle projektu jsou následující: najít biologické (genomické, proteomické a cytomické) faktory progrese z MGUS do MM; zastavit progresi do MM u nemocných s vysoce rizikovou MGUS pomocí experimentální léčby; informovat správně osoby s MGUS o riziku progrese do nádorového onemocnění podle typu individuálního rizika. Vytvoření nového a užitečnějšího modelu, než jsou stávající modely umožňující identifi kaci nemocných s vysoce rizikovým MGUS, je hlavním cílem tohoto mezinárodního projektu.

Monoclonal gammopathy of undertermined signifi cance (MGUS) is precancerosis mainly of multiple myeloma (MM). Project MGUS 2010 is the fi rst project generated from Register of Monoclonal Gammopathy (RMG; activated in 2007) of Czech Myeloma Group, myeloma section of Czech Hematology Society respectively. Main aims of this project are: to fi nd biological (genomic/proteomic/cytomic) markers of progression from MGUS to MM); to stop progression form high risk MGUS to MM using experimental therapy; to inform precisely people with MGUS about risk of progression. Creation of a new model superior to currently used systems for the identifi cation of high-risk MGUS patients is the target outcome of this international project.

• MeSH
• biologické markery analýza   MeSH
• financování organizované MeSH
• hodnocení rizik MeSH
• lidé MeSH
• monoklonální gamapatie nejasného významu klasifikace   MeSH
• myelomové proteiny moč   MeSH
• prekancerózy diagnóza   MeSH
• registrace MeSH
• rizikové faktory MeSH
• Check Tag
• lidé MeSH

Proposed project will develop a technological platform for investigation of complex suspension tissues (peripheral blood, bone marrow and CUSA aspirates) that contain a tumor infiltration. The approach would combine and complement two innovative high-content approaches: a single cell mass cytometry (“cancer cytomics”) to determine changes in cellular composition of tumor affected tissue and SEC-MAP array (“proteomics”) to determine changes in proteome of tumor cells. First, a reference of normal cellular development will be established for bone marrow resident B-cells to which abnormal (leukemic) cells could be compared in search for biomarkers. Next, SEC-MAP will be used to determine tumor specific proteome changes (in leukemia and in brain tumors as a model). Last, useful biomarkers will be built into single cells analysis by mass cytometry panels for “cancer cytomics” investigation. Bioinformatic tools will be developed to enable understanding the complex data.

Projekt vytvoří technologickou platformu pro výzkum komplexních tekutých tkání (periferní krev, kostní dřeň a CUSA aspirát) které jsou infiltrovány nádorovými buňkami. Přístup kombinuje dvě inovativní datově bohaté technologie: hmotnostní cytometrii („nádorová cytomika“), která zjišťuje změny v buněčném zastoupení infiltrovaných tkání a SEC-MAP array („proteomika“), která zjišťuje změny v proteomu nádorových buněk. Nejprve zmapujeme normální vývoj B-lymfocytů rezidentních v kostní dření, ke kterému bude možné vztahovat nádorem způsobené abnormity (potenciální biomarkery). Na modelu leukemických buněk a buněk mozkových nádorů dále určíme změny proteomu při maligním zvratu metodou SEC-MAP arrays. V další fázi budou biomarkery zabudované do panelů vyšetřovaných hmotnostní cytometrií („nádorová cytomika“). Vyvineme bioinformatické nástroje pro porozumění vzniklým vysoce kompexním datům.

• MeSH
• B-lymfocyty cytologie   MeSH
• cytodiagnostika metody   MeSH
• hmotnostní spektrometrie metody   MeSH
• leukemie MeSH
• nádorová transformace buněk MeSH
• nádorové biomarkery MeSH
• nádorové mikroprostředí MeSH
• nádory centrálního nervového systému MeSH
• nádory diagnóza   MeSH
• proteomika metody   MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• onkologie
• cytologie, klinická cytologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

Psoriasis vulgaris (PV) is a chronic, recurrent inflammatory dermatosis mediated by aberrantly activated immune cells. The role of the innate-like T cells, particularly gammadelta T (γδT) cells and MR1-restricted T lymphocytes, is incompletely explored, mainly through animal models, or by use of surrogate lineage markers, respectively. Here, we used case-control settings, multiparameter flow cytometry, 5-OP-RU-loaded MR1-tetramers, Luminex technology and targeted qRT-PCR to dissect the cellular and transcriptional landscape of γδ and MR1-restricted blood T cells in untreated PV cases (n=21, 22 matched controls). High interpersonal differences in cell composition were observed, fueling transcriptional variability at healthy baseline. A minor subset of canonical CD4+CD8+MR1-tet+TCRVα7.2+ and CD4+CD8-MR1-tet+TCRVα7.2+ T cells was the most significantly underrepresented community in male PV individuals, whereas Vδ2+ γδ T cells expressing high levels of TCR and Vδ1-δ2- γδ T cells expressing intermediate levels of TCR were selectively enriched in affected males, partly reflecting disease severity. Our findings highlight a formerly unappreciated skewing of human circulating MAIT and γδ cytomes during PV, and reveal their compositional changes in relation to sex, CMV exposure, serum cytokine content, BMI, and inflammatory burden. Complementing numerical alterations, we finally show that flow-sorted, MAIT and γδ populations exhibit divergent transcriptional changes in mild type I psoriasis, consisting of differential bulk expression for signatures of cytotoxicity/type-1 immunity (EOMES, RUNX3, IL18R), type-3 immunity (RORC, CCR6), and T cell innateness (ZBTB16).

• MeSH
• buněčná diferenciace MeSH
• cytotoxicita imunologická MeSH
• dospělí MeSH
• histokompatibilita - antigeny třídy I metabolismus   MeSH
• krevní oběh MeSH
• lidé středního věku MeSH
• lidé MeSH
• MAIT buňky imunologie   MeSH
• mladý dospělý MeSH
• přirozená imunita MeSH
• protein promyelocytické leukemie s motivem zinkového prstu genetika   metabolismus   MeSH
• psoriáza imunologie   MeSH
• receptory antigenů T-buněk gama-delta metabolismus   MeSH
• T-lymfocyty imunologie   MeSH
• Th1 buňky imunologie   MeSH
• vedlejší histokompatibilní antigeny metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH