Air pollution caused by road traffic has an unfavorable impact on the environment and also on human health. It has previously been shown, that complete gasoline emissions lead to toxic effects in cell models originating from human airways. Here we focused on extractable organic matter (EOM) from particulate matter, collected from gasoline emissions from fuels with different ethanol content. We performed cytotoxicity evaluation, quantification of mucin and extracellular reactive oxygen species (ROS) production, DNA breaks detection, and selected gene deregulation analysis, after one and five days of exposure of human bronchial epithelial model (BEAS-2B) and a 3D model of the human airway (MucilAirTM). Our data suggest that the longer exposure had more pronounced effects on the parameters of cytotoxicity and mucin production, while the impacts on ROS generation and DNA integrity were limited. In both cell models the expression of CYP1A1 was induced, regardless of the exposure period or EOM tested. Several other genes, including FMO2, IL1A, or TNF, were deregulated depending on the exposure time. In conclusion, ethanol content in the fuels did not significantly impact the toxicity of EOM. Biological effects were mostly linked to xenobiotics metabolism and inflammatory response. BEAS-2B cells were more sensitive to the treatment.

• MeSH
• benzin * MeSH
• bronchy cytologie   MeSH
• buněčné linie MeSH
• cytochrom P-450 CYP1A1 genetika   MeSH
• epitelové buňky účinky léků   metabolismus   MeSH
• histony metabolismus   MeSH
• interleukin-1alfa genetika   MeSH
• látky znečišťující vzduch toxicita   MeSH
• lidé MeSH
• oxygenasy genetika   MeSH
• pevné částice toxicita   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• regulace genové exprese účinky léků   MeSH
• TNF-alfa genetika   MeSH
• výfukové emise vozidel toxicita   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Polycyclic aromatic hydrocarbons (PAHs) associated with particulate matter (PM) may induce oxidative damage via reactive oxygen species (ROS) generation. However, the kinetics of ROS production and the link with antioxidant response induction has not been well studied. To elucidate the differences in oxidative potential of individual PAH compounds and extractable organic matter (EOM) from PM containing various PAH mixtures, we studied ROS formation and antioxidant response [total antioxidant capacity (TAC) and expression of HMOX1 and TXNRD1] in human alveolar basal epithelial cells (A549 cells) and human embryonic lung fibroblasts (HEL12469 cells). We treated the cells with three concentrations of model PAHs (benzo[a]pyrene, B[a]P; 3-nitrobenzanthrone, 3-NBA) and EOM from PM <2.5 μm (PM2.5). ROS levels were evaluated at 8 time intervals (30 min-24 h). In both cell lines, B[a]P treatment was associated with a time-dependent decrease of ROS levels. This trend was more pronounced in HEL12469 cells and was accompanied by increased TAC. A similar response was observed upon 3-NBA treatment in HEL12469 cells. In A549 cells, however, this compound significantly increased superoxide levels. This response was accompanied by the decrease of TAC as well as HMOX1 and TXNRD1 expression. In both cell lines, a short-time exposure to EOMs tended to increase ROS levels, while a marked decrease was observed after longer treatment periods. This was accompanied by the induction of HMOX1 and TXNRD1 expression in HEL12469 cells and increased TAC in A549 cells. In summary, our data indicate that in the studied cell lines B[a]P and EOMs caused a time-dependent decrease of intracellular ROS levels, probably due to the activation of the antioxidant response. This response was not detected in A549 cells following 3-NBA treatment, which acted as a strong superoxide inducer. Pro-oxidant properties of EOMs are limited to short-time exposure periods.

• MeSH
• buňky A549 MeSH
• fibroblasty účinky léků   metabolismus   patologie   MeSH
• kinetika MeSH
• kultivované buňky MeSH
• lidé MeSH
• organické látky škodlivé účinky   MeSH
• pevné částice škodlivé účinky   MeSH
• plíce účinky léků   metabolismus   patologie   MeSH
• polycyklické aromatické uhlovodíky škodlivé účinky   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

We investigated the role of oxidative damage in the mechanism of action of selected individual carcinogenic PAHs (c-PAHs: benzo[a]pyrene, B[a]P; dibenzo[a,l]pyrene, DB[a,l]P), an artificial mixture of c-PAHs (c-PAHs mix) and extractable organic matter (EOM) from urban air particulate matter (PM). Two cell lines (human hepatoma cells, HepG2; human diploid lung fibroblasts, HEL) were treated for 24 and 48h with various concentrations of compounds and mixtures. A panel of oxidative stress markers included 8-oxodeoxyguanosine (8-oxodG), 15-F(2t)-isoprostane (15-F(2t)-IsoP) and protein carbonyl groups. The response of the cell lines to the test compounds was substantially different. In HepG2 cells, oxidative damage to DNA was generally not induced by individual c-PAHs and the c-PAHs mix, but EOM increased 8-oxodG levels in these cells. In HEL cells, none of the compounds induced oxidative DNA damage. Lipid peroxidation, measured as the level of 15-F(2t)-IsoP, was induced by c-PAHs in HepG2 cells only after 48h of incubation, while the effect of EOM was detected already after 24h. In HEL cells, individual c-PAHs and the c-PAH mix generally decreased 15-F(2t)-IsoP levels. This effect was even stronger for EOM treatment. Protein oxidation, assessed as carbonyl levels in cell lysates, was not induced after 24h of treatment with any compound in either cell line. Individual c-PAHs and the c-PAH mix generally induced protein oxidation in both cell lines after 48h treatment, with the exception of DB[a,l]P in HepG2 cells. Oxidative damage to proteins caused by EOM was generally increased in HepG2 cells after 48h of incubation, while in HEL cells the effect was observed for only one dose of EOM. In summary, our results demonstrate the ability of EOM to induce oxidative damage to DNA and lipids after 24h of treatment, and to proteins after 48h, in HepG2 cells, while the effect of c-PAHs was substantially less. The induction of oxidative stress by c-PAHs and EOM in HEL cells was weak.

• MeSH
• buněčné linie MeSH
• buňky Hep G2 MeSH
• karcinogeny životního prostředí toxicita   MeSH
• látky znečišťující vzduch toxicita   MeSH
• lidé MeSH
• oxidace-redukce MeSH
• oxidační stres účinky léků   MeSH
• peroxidace lipidů účinky léků   MeSH
• pevné částice toxicita   MeSH
• plíce účinky léků   MeSH
• polycyklické aromatické uhlovodíky toxicita   MeSH
• poškození DNA účinky léků   MeSH
• proteiny účinky léků   MeSH
• zdraví ve městech MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Polycyclic aromatic hydrocarbons (PAHs) may cause lipid peroxidation via reactive oxygen species generation. 15-F2t-isoprostane (IsoP), an oxidative stress marker, is formed from arachidonic acid (AA) by a free-radical induced oxidation. AA may also be converted to prostaglandins (PG) by prostaglandin-endoperoxide synthase (PTGS) induced by NF-κB. We treated human embryonic lung fibroblasts (HEL12469) with benzo[a]pyrene (B[a]P), 3-nitrobenzanthrone (3-NBA) and extractable organic matter (EOM) from ambient air particulate matter <2.5 µm for 4 and 24 h. B[a]P and 3-NBA induced expression of PAH metabolising, but not antioxidant enzymes. The concentrations of IsoP decreased, whereas the levels of AA tended to increase. Although the activity of NF-κB was not detected, the tested compounds affected the expression of prostaglandin-endoperoxide synthase 2 (PTGS2). The levels of prostaglandin E2 (PGE2) decreased following exposure to B[a]P, whereas 3-NBA exposure tended to increase PGE2 concentration. A distinct response was observed after EOM exposure: expression of PAH-metabolising enzymes was induced, IsoP levels increased after 24-h treatment but AA concentration was not affected. The activity of NF-κB increased after both exposure periods, and a significant induction of PTGS2 expression was found following 4-h treatment. Similarly to PAHs, the EOM exposure was associated with a decrease of PGE2 levels. In summary, exposure to PAHs with low pro-oxidant potential results in a decrease of IsoP levels implying 'antioxidant' properties. For such compounds, IsoP may not be a suitable marker of lipid peroxidation.

• MeSH
• aromatické hydroxylasy metabolismus   MeSH
• benz(a)anthraceny toxicita   MeSH
• benzopyren toxicita   MeSH
• cyklooxygenasa 2 metabolismus   MeSH
• dinoprost analogy a deriváty   biosyntéza   metabolismus   MeSH
• dinoproston biosyntéza   metabolismus   MeSH
• fibroblasty účinky léků   enzymologie   MeSH
• kultivované buňky MeSH
• kyselina arachidonová metabolismus   MeSH
• látky znečišťující vzduch toxicita   MeSH
• lidé MeSH
• NF-kappa B metabolismus   MeSH
• oxidační stres účinky léků   MeSH
• peroxidace lipidů účinky léků   MeSH
• pevné částice toxicita   MeSH
• plíce cytologie   účinky léků   embryologie   enzymologie   MeSH
• polycyklické aromatické uhlovodíky toxicita   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Many of the toxic and carcinogenic effects of urban air pollution have been linked to polycyclic aromatic hydrocarbons (PAHs) adsorbed to airborne particulate matter (PM). The carcinogenic properties of PAHs in complex organic mixtures derived from PM have been chiefly attributed to their mutagenicity. Nevertheless, PAHs are also potent activators of the aryl hydrocarbon receptor (AhR), which may contribute to their nongenotoxic effects, including tumor promotion. As the genotoxicity of carcinogenic PAHs in complex mixtures derived from urban PM is often inhibited by other mixture constituents, the AhR-mediated activity of urban PM extracts might significantly contribute to the carcinogenic activity of such mixtures. In the present study, we used an organic extract of the urban dust standard reference material, SRM1649a, as a model mixture to study a range of toxic effects related to DNA damage and AhR activation. Both the organic extract and its neutral aromatic fraction formed a low number of DNA adducts per nucleotide in the liver epithelial WB-F344 cells model, without inducing DNA damage response, such as tumor suppressor p53 activation and apoptosis. In contrast, we found that this extract, as well as its neutral and polar fractions, were potent inducers of a range of AhR-mediated responses, including induction of the AhR-mediated transcription, such as cytochrome P450 1A1/1B1 expression, and the AhR-dependent cell proliferation. Importantly, these toxic events occurred at doses one order of magnitude lower than DNA damage. The AhR-mediated activity of the neutral fraction was linked to PAHs and their derivatives, as polychlorinated dibenzo-p-dioxins, dibenzofurans and biphenyls were only minor contributors to the overall AhR-mediated activity. Taken together, our data suggest that more attention should be paid to the AhR-dependent nongenotoxic events elicited by urban PM constituents, especially PAHs and their derivatives.

• MeSH
• adukty DNA účinky léků   MeSH
• apoptóza účinky léků   MeSH
• buněčné linie MeSH
• cytochrom P-450 CYP1A1 metabolismus   MeSH
• geny p53 účinky léků   MeSH
• játra účinky léků   MeSH
• krysa rodu rattus MeSH
• mutageny toxicita   MeSH
• organické látky toxicita   MeSH
• pevné částice toxicita   MeSH
• polycyklické aromatické uhlovodíky toxicita   MeSH
• poškození DNA účinky léků   MeSH
• proliferace buněk účinky léků   MeSH
• receptory aromatických uhlovodíků metabolismus   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

A rapid and reliable analytical method was developed for the quantitative determination of psychopharmaceuticals, their precursors and by-products in real contaminated samples from a pharmaceutical company in Olomouc (Czech Republic), based on SPE disk extraction and detection by ultra performance liquid chromatography, combined with time-of-flight mass spectrometry. The target compounds were quantified in the real whole-water samples (water including suspended particles), both in the presence of suspended particulate matter (SPM) and high concentrations of other organic pollutants. A total of nine compounds were analyzed which consisted of three commonly used antidepressants (tricyclic antidepressants and antipsychotics), one antitussive agent and five by-products or precursors. At first, the SPE disk method was developed for the extraction of water samples (dissolved analytes, recovery 84-104%) and pressurised liquid extraction technique was verified for solid matrices (sludge samples, recovery 81-95%). In order to evaluate the SPE disk technique for whole water samples containing SPM, non contaminated groundwater samples were also loaded with different amounts (100 and 300mgL(-1)) of real contaminated sludge originating from the same locality. The recoveries from the whole-water samples obtained by SPE disk method ranged between 67 and 119% after the addition of the most contaminated sludge. The final method was applied to several real groundwater (whole-water) samples from the industrial area and high concentrations (up to 10(3)μgL(-1)) of the target compounds were detected. The results of this study document and indicate the feasibility of the SPE disk method for analysis of groundwater.

• MeSH
• chemické látky znečišťující vodu analýza   MeSH
• extrakce na pevné fázi * MeSH
• hmotnostní spektrometrie * MeSH
• monitorování životního prostředí přístrojové vybavení   metody   MeSH
• odpadní voda chemie   MeSH
• pevné částice analýza   MeSH
• podzemní voda chemie   MeSH
• psychotropní léky analýza   MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

BACKGROUND: Recently, we used cell-free assays to demonstrate the toxic effects of complex mixtures of organic extracts from urban air particles (PM2.5) collected in four localities of the Czech Republic (Ostrava-Bartovice, Ostrava-Poruba, Karvina and Trebon) which differed in the extent and sources of air pollution. To obtain further insight into the biological mechanisms of action of the extractable organic matter (EOM) from ambient air particles, human embryonic lung fibroblasts (HEL12469) were treated with the same four EOMs to assess changes in the genome-wide expression profiles compared to DMSO treated controls. METHOD: For this purpose, HEL cells were incubated with subtoxic EOM concentrations of 10, 30, and 60 μg EOM/ml for 24 hours and global gene expression changes were analyzed using human whole genome microarrays (Illumina). The expression of selected genes was verified by quantitative real-time PCR. RESULTS: Dose-dependent increases in the number of significantly deregulated transcripts as well as dose-response relationships in the levels of individual transcripts were observed. The transcriptomic data did not differ substantially between the localities, suggesting that the air pollution originating mainly from various sources may have similar biological effects. This was further confirmed by the analysis of deregulated pathways and by identification of the most contributing gene modulations. The number of significantly deregulated KEGG pathways, as identified by Goeman's global test, varied, depending on the locality, between 12 to 29. The Metabolism of xenobiotics by cytochrome P450 exhibited the strongest upregulation in all 4 localities and CYP1B1 had a major contribution to the upregulation of this pathway. Other important deregulated pathways in all 4 localities were ABC transporters (involved in the translocation of exogenous and endogenous metabolites across membranes and DNA repair), the Wnt and TGF-β signaling pathways (associated particularly with tumor promotion and progression), Steroid hormone biosynthesis (involved in the endocrine-disrupting activity of chemicals), and Glycerolipid metabolism (pathways involving the lipids with a glycerol backbone including lipid signaling molecules). CONCLUSION: The microarray data suggested a prominent role of activation of aryl hydrocarbon receptor-dependent gene expression.

• MeSH
• exprese genu účinky léků   MeSH
• fibroblasty cytologie   účinky léků   fyziologie   MeSH
• látky znečišťující vzduch chemie   metabolismus   farmakologie   MeSH
• lidé MeSH
• mikročipová analýza MeSH
• organické látky chemie   metabolismus   farmakologie   MeSH
• oxidace-redukce MeSH
• pevné částice chemie   metabolismus   farmakologie   MeSH
• plíce cytologie   MeSH
• stanovení celkové genové exprese MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

The main aim of this study was to compare the genotoxic potential of organic extracts from urban air particles collected in three different sampling periods in the center of Prague (Czech Republic). For this purpose, we analyzed the DNA adduct forming activity of extractable organic matter (EOM) from urban air particles <10 microm (PM10) in the human hepatoma cell line HepG2. DNA adducts were analyzed by (32)P-postlabelling with nuclease P1 enrichment. PM10 concentrations were 36.9 microg/m(3), 62.6mug/m(3) and 39.0 microg/m(3), in summer 2000, winter 2001 and winter 2005, respectively. The corresponding EOM contents were 5.0 microg/m(3) (13.9% of PM10), 14.9 microg/m(3) (23.8%) and 6.7 microg/m(3) (17.2%). The total DNA adduct levels induced by 10 microg EOM/ml were 4.7, 19.5 and 37.2 adducts/10(8) nucleotides in summer 2000, winter 2001 and winter 2005, respectively. However, when the EOM quantities per cubic meter of air were taken into consideration, the summer sample exhibited a 10-fold lower genotoxicity than did those of winter, while the difference between the winter samples was not significant: 23.4 in summer 2000, 291 in winter 2001 and 249 in winter 2005 (in relative units). Although the PM10 concentration in air and the EOM content in particles in winter 2005 were significantly lower than in winter 2001, the genotoxic potential of the ambient air in these samples was almost equal. There were significant positive correlations between the B[a]P and c-PAH content in EOM from various sampling periods and the total DNA adduct levels detected in the EOM-treated samples. These findings support the hypothesis that the B[a]P and c-PAH content in EOM is the most important factor that determines its genotoxic potential. Thus, estimating the genotoxic potential of the ambient air and predicting health risk should be based mainly on the c-PAH concentration and the biological activity of the extracts, while the mass of particles and the EOM content do not seem to be crucial determinants of ambient air genotoxicity.

• MeSH
• adukty DNA metabolismus   účinky léků   MeSH
• chromatografie na tenké vrstvě MeSH
• financování organizované MeSH
• izotopy fosforu MeSH
• látky znečišťující vzduch analýza   chemie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• pevné částice analýza   farmakologie   MeSH
• polycyklické aromatické uhlovodíky analýza   farmakologie   MeSH
• roční období MeSH
• velkoměsta MeSH
• Check Tag
• lidé MeSH
• Geografické názvy
• velkoměsta MeSH

Principal aims of this study were at first, to find a relevant human derived cell line to investigate the genotoxic potential of PAH-containing complex mixtures and second, to use this cell system for the analysis of DNA adduct forming activity of organic compounds bound onto PM10 particles. Particles were collected by high volume air samplers during summer and winter periods in three European cities (Prague, Kosice, and Sofia), representing different levels of air pollution. The genotoxic potential of extractable organic matter (EOM) was compared with the genotoxic potential of individual carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) as well as their artificial mixtures. Metabolically competent human hepatoma HepG2 cells, confluent cultures of human diploid lung fibroblasts (HEL), and the human monocytic leukemia cell line THP-1 were used as models. DNA adducts were analyzed by (32)P-postlabeling. The total DNA adduct levels induced in HepG2 cells after exposure to EOMs were higher than in HEL cells treated under the same conditions (15-190 versus 2-15adducts/10(8) nucleotides, in HepG2 and HEL cells, respectively). THP-1 cells exhibited the lowest DNA adduct forming activity induced by EOMs (1.5-3.7adducts/10(8) nucleotides). A direct correlation between total DNA adduct levels and c-PAH content in EOM was found for all EOMs in HepG2 cells incubated with 50microg EOM/ml (R=0.88; p=0.0192). This correlation was even slightly stronger when B[a]P content in EOMs and B[a]P-like adduct spots were analyzed (R=0.90; p=0.016). As THP-1 cells possess a limited metabolic capacity for most c-PAHs to form DNA reactive intermediates and are also more susceptible to toxic effects of PAHs and various EOM components, this cell line seemed to be an inappropriate system for genotoxicity studies of PAH-containing complex mixtures. The seasonal variability of genotoxic potential of extracts was stronger than variability among the three localities studied. In HepG2 cells, the highest DNA adduct levels were induced by EOM collected in Prague in the winter period, followed by Sofia and Kosice. However, in the summer sampling period, the order was quite opposite: Kosice>Sofia>Prague. When the EOM content per m(3) of air was taken into consideration in order to compare real exposures of humans to genotoxic compounds in all three localities, extracts from respirable dust particles collected in Sofia exhibited the highest genotoxicity regardless of the sampling period. The results indicate that most of DNA adducts detected in cells incubated with EOMs have their origin in low concentrations of c-PAHs representing 0.03-0.17% of EOM total mass. Finally, our results suggest that HepG2 cells have a metabolic capacity for PAHs similar to human hepatocytes and represent therefore the best in vitro model for investigating the genotoxic potential of complex mixtures containing PAHs among the three cell lines tested in this study.

• MeSH
• adukty DNA analýza   MeSH
• financování organizované MeSH
• karcinogeny životního prostředí toxicita   MeSH
• látky znečišťující vzduch toxicita   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• organické látky toxicita   MeSH
• pevné částice toxicita   MeSH
• polycyklické aromatické uhlovodíky metabolismus   toxicita   MeSH
• testy genotoxicity metody   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH

The genotoxic activities of complex mixtures of organic extracts from the urban air particles collected in various localities of the Czech Republic, which differed in the extent and sources of air pollution, were compared. For this purpose, PM2.5 particles were collected by high volume samplers in the most polluted area of the Czech Republic--Ostrava region (localities Bartovice, Poruba and Karvina) and in the locality exhibiting a low level of air pollution--Trebon--a small town in the non-industrial region of Southern Bohemia. To prepare extractable organic matter (EOM), PM2.5 particles were extracted by dichloromethane and c-PAHs contents in the EOMs were determined. As markers of genotoxic potential, DNA adduct levels and oxidative DNA damage (8-oxo-7,8-dihydro-2'-deoxyguanosine, 8-oxodG, levels) induced by EOMs in an acellular assay of calf thymus DNA coupled with ³²P-postlabeling (DNA adducts) and ELISA (8-oxodG) in the presence and absence of microsomal S9 fraction were employed. Twofold higher DNA adduct levels (17.20 adducts/10⁸ nucleotides/m³ vs. 8.49 adducts/10⁸ nucleotides/m³) were induced by EOM from Ostrava-Bartovice (immediate proximity of heavy industry) compared with that from Ostrava-Poruba (mostly traffic emissions). Oxidative DNA damage induced by EOM from Ostrava-Bartovice was more than fourfold higher than damage induced by EOM from Trebon (8-oxodG/10⁸ dG/m³: 0.131 vs. 0.030 for Ostrava-Bartovice vs. Trebon, respectively). Since PM2.5 particles collected in various localities differ with respect to their c-PAHs content, and c-PAHs significantly contribute to genotoxicity (DNA adduct levels), we suggest that monitoring of PM2.5 levels is not a sufficient basis to assess genotoxicity of respirable aerosols. It seems likely that the industrial emissions prevailing in Ostrava-Bartovice represent a substantially higher genotoxic risk than mostly traffic-related emissions in Ostrava-Poruba. B[a]P and c-PAH contents in EOMs are the most important factors relating to their genotoxic potential.

• MeSH
• adukty DNA analýza   MeSH
• deoxyguanosin analogy a deriváty   analýza   MeSH
• DNA účinky léků   MeSH
• karcinogeny životního prostředí chemie   toxicita   MeSH
• látky znečišťující vzduch chemie   toxicita   MeSH
• organické látky chemie   toxicita   MeSH
• oxidační stres MeSH
• pevné částice chemie   toxicita   MeSH
• polycyklické aromatické uhlovodíky analýza   MeSH
• poškození DNA MeSH
• průmyslový odpad škodlivé účinky   analýza   MeSH
• skot MeSH
• testy genotoxicity MeSH
• výfukové emise vozidel analýza   toxicita   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH