In vitro kultivace motolice Trichobilharzia regenti

Třída Trematoda zahrnuje mnoho patogenních zástupců, mezi nimi významného lidského parazita druhu Schistosoma mansoni (čeleď Schistosomatidae). Její blízký příbuzný, ptačí schistosoma Trichobilharzia regenti, je hlavním předmětem této práce. In vitro kultivace motolic může být vhodným krokem pro bližší pochopení biologie těchto organizmů a jejich vztahů s hostitelem. Žádný druh motolice však dosud nebyl v in vitro podmínkách kultivován od vajíčka po dospělce produkujícího opět vajíčka. Kultivací S. mansoni se do dnešní doby zabývalo mnoho studií, ale informace v literatuře týkající se kultivace T. regenti jsou prozatím strohé. V tomto ohledu bylo dosud pouze dosaženo in vitro transformace cerkárií T. regenti ve schistosomula s následným přežitím několik dní v kultivačním médiu. S cílem získat co nejvyšší množství cerkárií s odlomeným ocáskem (jakožto stimulem pro transformaci cerkárií T. regenti ve schistosomula) bylo porovnáno osm transformačních metod. Vybraných pět metod bylo dále zhodnoceno z hlediska základních charakteristik časného schistosomula (svlečený glykokalyx, vyprázdněné penetrační žlázy, přežití v in vitro podmínkách). Bylo zjištěno, že transformace cerkárií s použitím injekční jehly či tkáňového homogenizátoru BeadBeateru poskytuje nejvyšší procento cerkárií s odlomeným ocáskem....The class Trematoda includes many pathogenic representatives. Main subject of this thesis, avian schistosome Trichobilharzia regenti, is a close relative to the important human pathogen Schistosoma mansoni (family Schistosomatidae). In vitro cultivation of trematodes enables closer understanding of their biology and parasite- host interactions; however, no trematode species has been successfully kept in vitro from the egg stage to the adults producing eggs. Many studies are focused on the problematic of S. mansoni cultivation, but data concerning T. regenti cultivation remain scarce. Only the ability of T. regenti cercariae to transform into schistosomula in vitro was documented, with following survival in a culture medium for a few days. Comparison of eight transformation methods was performed with T. regenti cercariae. Based on the number of tailless cercarial bodies obtained, five transformation methods were selected for further evaluation of the early schistosomula characteristics (glycocalyx shedding, penetration glands emptying and survival in vitro). It was observed that the largest quantity of cercarial bodies can be obtained by using a syringe needle or the BeadBeater cell disrupter. The largest quantity of schistosomula meeting the criteria of early schistosomulum was recorded after...Department of ParasitologyKatedra parazitologiePřírodovědecká fakultaFaculty of Scienc

TREATMENT OF NATURAL GAS BY ADSORPTION OF CO2

Apart from burning, one of the possible uses of natural gas is as a fuel for motor vehicles. There are two types of fuel from natural gas — CNG (Compressed Natural Gas) or LNG (Liquefied Natural Gas). Liquefaction of natural gas is carried out for transport by tankers, which are an alternative to long-distance gas pipelines, as well as for transport over short distance, using LNG as a fuel for motor vehicles. A gas adjustment is necessary to get LNG. As an important part of the necessary adjustment of natural gas to get LNG, a reduction of CO2 is needed. There is a danger of the carbon dioxide freezing during the gas cooling. This work deals with the testing of adsorption removal of CO2 from natural gas. The aim of these measurements was to find a suitable adsorbent for CO2 removal from natural gas. Two different types of adsorbents were tested: activated carbon and molecular sieve. The adsorption properties of the selected adsorbents were tested and compared. The breakthrough curves for CO2 for both adsorbents were measured. The conditions of the testing were estimated according to conditions at a gas regulation station — 4.0MPa pressure and 8 °C temperature. Natural gas was simulated by model gas mixture during the tests. The breakthrough volume was set as the gas volume passing through the adsorber up to the CO2 concentration of 300 ml/m3 in the exhaust gas. The thermal and pressure desorption of CO2 from saturated adsorbents were also tested after the adsorption

TREATMENT OF NATURAL GAS BY ADSORPTION OF CO2

Apart from burning, one of the possible uses of natural gas is as a fuel for motor vehicles. There are two types of fuel from natural gas — CNG (Compressed Natural Gas) or LNG (Liquefied Natural Gas). Liquefaction of natural gas is carried out for transport by tankers, which are an alternative to long-distance gas pipelines, as well as for transport over short distance, using LNG as a fuel for motor vehicles. A gas adjustment is necessary to get LNG. As an important part of the necessary adjustment of natural gas to get LNG, a reduction of CO2 is needed. There is a danger of the carbon dioxide freezing during the gas cooling. This work deals with the testing of adsorption removal of CO2 from natural gas. The aim of these measurements was to find a suitable adsorbent for CO2 removal from natural gas. Two different types of adsorbents were tested: activated carbon and molecular sieve. The adsorption properties of the selected adsorbents were tested and compared. The breakthrough curves for CO2 for both adsorbents were measured. The conditions of the testing were estimated according to conditions at a gas regulation station — 4.0MPa pressure and 8 °C temperature. Natural gas was simulated by model gas mixture during the tests. The breakthrough volume was set as the gas volume passing through the adsorber up to the CO2 concentration of 300 ml/m3 in the exhaust gas. The thermal and pressure desorption of CO2 from saturated adsorbents were also tested after the adsorption

Comparative analysis of toxic responses of organic extracts from diesel and selected alternative fuels engine emissions in human lung BEAS-2B cells

This study used toxicogenomics to identify the complex biological response of human lung BEAS-2B cells treated with organic components of particulate matter in the exhaust of a diesel engine. First, we characterized particles from standard diesel (B0), biodiesel (methylesters of rapeseed oil) in its neat form (B100) and 30% by volume blend with diesel fuel (B30), and neat hydrotreated vegetable oil (NEXBTL100). The concentration of polycyclic aromatic hydrocarbons (PAHs) and their derivatives in organic extracts was the lowest for NEXBTL100 and higher for biodiesel. We further analyzed global gene expression changes in BEAS-2B cells following 4 h and 24 h treatment with extracts. The concentrations of 50 mu g extract/mL induced a similar molecular response. The common processes induced after 4 h treatment included antioxidant defense, metabolism of xenobiotics and lipids, suppression of pro-apoptotic stimuli, or induction of plasminogen activating cascade; 24 h treatment affected fewer processes, particularly those involved in detoxification of xenobiotics, including PAHs. The majority of distinctively deregulated genes detected after both 4 h and 24 h treatment were induced by NEXBTL100; the deregulated genes included, e.g., those involved in antioxidant defense and cell cycle regulation and proliferation. B100 extract, with the highest PAH concentrations, additionally affected several cell cycle regulatory genes and p38 signaling.Web of Science1711art. no. 183

DNA methylation profiles in a group of workers occupationally exposed to nanoparticles

The risk of exposure to nanoparticles (NPs) has rapidly increased during the last decade due to the vast use of nanomaterials (NMs) in many areas of human life. Despite this fact, human biomonitoring studies focused on the effect of NP exposure on DNA alterations are still rare. Furthermore, there are virtually no epigenetic data available. In this study, we investigated global and gene-specific DNA methylation profiles in a group of 20 long-term (mean 14.5 years) exposed, nanocomposite, research workers and in 20 controls. Both groups were sampled twice/day (pre-shift and post-shift) in September 2018. We applied Infinium Methylation Assay, using the Infinium MethylationEPIC BeadChips with more than 850,000 CpG loci, for identification of the DNA methylation pattern in the studied groups. Aerosol exposure monitoring, including two nanosized fractions, was also performed as proof of acute NP exposure. The obtained array data showed significant differences in methylation between the exposed and control groups related to long-term exposure, specifically 341 CpG loci were hypomethylated and 364 hypermethylated. The most significant CpG differences were mainly detected in genes involved in lipid metabolism, the immune system, lung functions, signaling pathways, cancer development and xenobiotic detoxification. In contrast, short-term acute NP exposure was not accompanied by DNA methylation changes. In summary, long-term (years) exposure to NP is associated with DNA epigenetic alterations

Inhalation of ZnO nanoparticles: Splice junction expression and alternative splicing in mice

Despite the wide application of nanomaterials, toxicity studies of nanoparticles (NP) are often limited to in vitro cell models, and the biological impact of NP exposure in mammals has not been thoroughly investigated. Zinc oxide (ZnO) NPs are commonly used in various consumer products. To evaluate the effects of the inhalation of ZnO NP in mice, we studied splice junction expression in the lungs as a proxy to gene expression changes analysis. Female ICR mice were treated with 6.46 x 10(4) and 1.93 x 10(6) NP/cm(3) for 3 days and 3 months, respectively. An analysis of differential expression and alternative splicing events in 298 targets (splice junctions) of 68 genes involved in the processes relevant to the biological effects of ZnO NP was conducted using next-generation sequencing. Three days of exposure resulted in the upregulation of IL-6 and downregulation of BID, GSR, NF-kB2, PTGS2, SLC11A2, and TXNRD1 splice junction expression; 3 months of exposure increased the expression of splice junctions in ALDH3A1, APAF1, BID, CASP3, DHCR7, GCLC, GCLM, GSR, GSS, EHHADH, FAS, HMOX-1, IFN, NF-kB1, NQO-1, PTGS1, PTGS2, RAD51, RIPK2, SRXN1, TRAF6, and TXNRD1. Alternative splicing of TRAF6 and TXNRD1 was induced after 3 days of exposure to 1.93 x 10(6) NP/cm(3). In summary, we observed changes of splice junction expression in genes involved in oxidative stress, apoptosis, immune response, inflammation, and DNA repair, as well as the induction of alternative splicing in genes associated with oxidative stress and inflammation. Our data indicate the potential negative biological effects of ZnO NP inhalation.Web of Science168120019

Genetic variability of dung beetles on pastures.

The aim of this thesis was a research of the genetic variability of 8 populations of the dung beetle Aphodius prodromus using allozyme markers. Following population-genetic analysis were used to determine genetic structure of these populations: estimates of heterozygosity, Wright´s F- statistics, estimates of gene flow, Mantel test, PCA, Nei´s genetic distance, testing of Hardy-Weinberg equilibrium and linkage disequilibrium. The results revealed low gene flow between populations, low number of heterozygotes and disequilibrium within and between populations. Cluster analyses showed, that populations from Tichá and Rychnov nad Malší-hřbitov are genetically most different from other populations

Cultivation of Trematodes

Many of Trematode species are serious pathogens of human and animals; therefore they are the subjects of intensive investigation. The successful in vitro cultivation of these organisms could represent the first step for further sophisticated experiments focused on characterization of crucial biological processes related to the particular phases of fluke life cycle or for the studies focused on testing the anthelmintic effect of various compounds. This thesis reviews majority of information published in relation to in vitro cultivation of Trematodes. It shows that for many Trematode families is our knowledge of culturing methods very limited. In contrast to the first attempts to cultivate some species of Trematodes about 100 years ago, nowadays we dispose of technologies enabling to cultivate particular live cells isolated from different tissues of flukes. Some of the cultivation media originally used for cell cultures are now commercially available and they were successfully used for cultivation of whole worms. Nevertheless, we still haven't been able to continuously cultivate a Trematode species by using an egg at the beginning and finish with fully developed adult producing eggs. However, there are some partially successful experiments in which development of particular life stages of some flukes has been..

Suicidal behaviour in adolescent age

The present thesis aims to concentrate available sources of scientific literature on adolescent suicidal behavior, which would also provide a tutu on certain issues holistic view. In the introductory part of the thesis points to the fact that suicidal behavior is being explored especially psychiatric disciplines because in psychiatric disorders are at high risk of suicidal behavior. The seriousness of this issue and the benefit of that statistical data on both the domestic and the World Health Organization, the number of suicide attempts and suicides are mentioned also the trends per decade for the Czech Republic (from r. 1990). In the next part of the thesis are presented in various forms of suicidal behavior with an indication of the usual motivation, quantitative data on the most frequently used methods of suicide in this country and abroad. In connection with the study of adolescent suicidal behavior are also explored in parallel manifestations of self-harm, stating the characteristic differences. The analysis of risk factors for suicidal behavior in children and adolescents. Special attention is paid to the influence of the family, the school environment, the role of their own state of health, religion, social situation, etc. Separately are analyzed negative effects of drug addiction. Attention is also..

In vitro cultivation of the trematode species Trichobilharzia regenti

The class Trematoda includes many pathogenic representatives. Main subject of this thesis, avian schistosome Trichobilharzia regenti, is a close relative to the important human pathogen Schistosoma mansoni (family Schistosomatidae). In vitro cultivation of trematodes enables closer understanding of their biology and parasite- host interactions; however, no trematode species has been successfully kept in vitro from the egg stage to the adults producing eggs. Many studies are focused on the problematic of S. mansoni cultivation, but data concerning T. regenti cultivation remain scarce. Only the ability of T. regenti cercariae to transform into schistosomula in vitro was documented, with following survival in a culture medium for a few days. Comparison of eight transformation methods was performed with T. regenti cercariae. Based on the number of tailless cercarial bodies obtained, five transformation methods were selected for further evaluation of the early schistosomula characteristics (glycocalyx shedding, penetration glands emptying and survival in vitro). It was observed that the largest quantity of cercarial bodies can be obtained by using a syringe needle or the BeadBeater cell disrupter. The largest quantity of schistosomula meeting the criteria of early schistosomulum was recorded after..