Výskyt antibiotik v životním prostředí

7 ABSTRAKT Univerzita Karlova v Praze, Farmaceutická fakulta v Hradci Králové Katedra analytické chemie Kandidát: Mgr. Marcela Seifrtová Školitel: Prof. RNDr. Petr Solich, CSc. Prof. Angelina Pena, Ph.D. Název disertační práce: Výskyt antibiotik v životním prostředí Výskyt léčiv a jejich metabolitů v životním prostředí je v posledních letech ve zvýšeném zájmu veřejnosti. Léčiva jsou látky určené mít speciální biologický účinek v organismu. Díky jejich fyzikálně-chemickým a biologickým vlastnostem mohou mít po uvolnění do životního prostředí důležitý vliv na lidské zdraví a na ekosystém. Léčiva nejsou považována za perzistentní látky z hlediska jejich poločasu, ale jsou neustále uvolňována do životního prostředí díky používání v humánní a veterinární medicíně. To má za následek dlouhodobou expozici pro vodní ekosystém. V současné době jsou léčiva považovány za velmi nebezpečné látky pro životní prostředí a je prováděn rozsáhlý výzkum týkající se jejich výskytu, osudu a efektů na životní prostředí a různé druhy organismů. Antibiotika jsou velmi významná skupina léčiv široce využívaná v humánní a veterinární medicíně k léčení různých druhů infekcí. Hlavní problém výskytu antibiotik v životním prostředí je spojen s rozvojem antibiotické rezistence. Předkládaná disertační práce je hlavně zaměřena na vývoj a...5 ABSTRACT Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Marcela Seifrtová Supervisor: Prof. RNDr. Petr Solich, CSc. Prof. Angelina Pena, Ph.D. Title of Doctoral Thesis: Occurrence of antibiotics in the environment The occurrence of pharmaceuticals in the environment is a growing concern. The pharmaceuticals are developed to have biological effects, and because of their biological and physico-chemical properties, their release into the environment can have impact in human health and ecosystems. Despite their long half-life, pharmaceuticals are constantly being release into the environment, contributing to a continuous exposure of aquatic ecosystems. The pharmaceuticals are considered as emerging environmental pollutants and the assessment of their environmental risk is acquired. Research on the occurrence, fate and effects is described in the scientific literature, however the data are still insufficient. Antibiotics are an important class of pharmaceuticals widely used in human and veterinary medicine. The main problem associated with their presence in the environment is the development of mechanisms of resistance by bacteria, leading to the development of multi-resistant bacteria, which may put at risk public health in terms of...Katedra analytické chemieDepartment of Analytical ChemistryFaculty of Pharmacy in Hradec KrálovéFarmaceutická fakulta v Hradci Králov

Precoloring extension in planar near-Eulerian-triangulations

We consider the 4-precoloring extension problem in planar near-Eulerian- triangulations, i.e., plane graphs where all faces except possibly for the outer one have length three, all vertices not incident with the outer face have even degree, and exactly the vertices incident with the outer face are precolored. We give a necessary topological condition for the precoloring to extend, and give a complete characterization when the outer face has length at most five and when all vertices of the outer face have odd degree and are colored using only three colors

Biosensor Applications in the Field of Antibiotic Research—A Review of Recent Developments

Antibacterials are among of the most important medications used in health care. However, their efficacy is increasingly impeded by a tremendous and globally spread bacterial resistance phenomenon. This bacterial resistance is accelerated by inadequate application of antibacterial drugs in humans, the widespread veterinary use of antibacterials, and antibacterial occurrence in the environment and food. Further, there is a lack of development of innovative novel drugs. Therefore, the search for novel antibacterials has to be intensified and the spread of antibacterials in the environment has to be restricted. Due to the fundamental progress in biosensor development and promising applications in the antibiotic field, this review gives for the first time an overview on the use and prospects of biosensor applications in that area. A number of reports have applied biosensors of different design and techniques to search for antibacterials in environmental and foodstuff matrices. These studies are discussed with respect to the analytical values and compared to conventional techniques. Furthermore, biosensor applications to elucidate the mode of action of antimicrobial drugs in vitro have been described. These studies were critically introduced referring to the informational value of those simulations. In summary, biosensors will be illustrated as an innovative and promising, although not yet comprehensively applied, technique in the antibacterial field

The molecular mechanisms of stem cells response to cytostatic treatment

The molecular mechanisms of stem cells response to cytostatic treatment Apoptosis has been suggested as a common mechanism by which the cells are eradicated during chemotherapy. Most studies have focused on the reaction to DNA damage in tumor cell lines, but it is obvious that not only the tumour cells, but also normal cells are exposed to DNA-damage inducing agents during an anticancer treatment. Within this study we focus on the reaction of stem cells which are an important component of tissues and represent promising tool for a tissue engineering. Before their use, however, their reaction to the DNA damage caused by various genotoxic stressors requires elucidation and characterization of these cells under genotoxic stress contributes to the evaluation of their safety usage. In this study we assessed the effects of the frequently used chemotherapeutic agents mitoxantrone (MTX) and cisplatin (CP) on mesenchymal stem cells isolated from a dental pulp (DPSCs) and compared it with the response of human dermal fibroblasts (HDFs). Our data indicate that both cell types react to MTX and CP by the induction of apoptosis as well as by a cell cycle arrest in a dose-dependent manner. DPSCs and HDFs treated with clinically relevant concentrations of MTX underwent a permanent cell cycle arrest called..

Regulární nakrytí - struktura a složitost

Regular Coverings - Structure and Complexity Michaela Seifrtová The thesis consists of two main parts, the first concentrated on the struc- ture of graph coverings, where different properties of regular graph coverings are presented, and the second dealing with computational complexity of the covering problem. Favorable results have been achieved in this area, proving the problem is solvable in polynomial time for all graphs whose order is a prime multiple of the order of the covered graph.

Regulations of p53 pathway in anticancer therapy: MAPK inhibitors

V této práci jsme se zabývali molekulárními změnami po působení mitoxantronu a inhibitoru U0126 (inhibitor MEK1/2) u lidských nádorových buněk. Cílem práce bylo zjistit, zda a jakým způsobem ovlivňuje inhibitor U0126 aktivaci biochemických cest po mitoxantronem vyvolaném poškození DNA. Jako modelový systém jsme použili buněčnou linii MOLT-4 odvozenou od lidské T-lymfocytární leukémie. Na počátku jsme sledovali proliferační aktivitu buněk po působení různých dávek mitoxantronu s cílem vybrat vhodné dávkové schéma pro další studii. Dále jsme sledovali proliferační aktivitu buněk po působení mitoxantronu a inhibitoru U0126 a hodnotili vliv tohoto inhibitoru na působení mitoxantronu. Zjistili jsme, že inhibice MEK1/2 kinázy pomocí U0126 vede po mitoxantronem vyvolaném poškození DNA ke zvýšenému odumírání buněk. Průtokovou cytometrií jsme sledovali indukci apoptózy za použití duálního značení Annexinem V a propidium jodidem a optických parametrů rozptylu světla. Indukci apoptózy jsme stanovili v reakci na různé dávky mitoxantronu, inhibitor U0126 a jejich kombinaci v různých časových intervalech působení. Bylo potvrzeno, že U0126 inhibicí MEK1/2 zastaví cestu napomáhající buněčné proliferaci a přežívání, a po poškození DNA je tak více buněk posíláno do apoptózy. Pro lepší přehled o průběhu buněčného cyklu jsme u jednotlivých skupin zanalyzovali obsah DNA v buňce. U buněk MOLT-4 mitoxantron způsobuje zástavu buněčného cyklu v G2/M fázi následovanou rozvojem apoptózy, inhibice MEK1/2 kinázy pomocí U0126 tuto zástavu buněčného cyklu nenarušuje. Následně jsme studovali změny v expresi proteinů podílejících se na regulaci buněčného cyklu a rozvoji apoptózy (p53 a jeho posttranslační modifikace, p21, ERK1/2, lamin B). Expresi proteinů jsme detekovali pomocí elektroforézy a Western blottingu s následnou imunodetekcí. Zjistili jsme, že inhibice MEK1/2 pomocí U0126 u buněk MOLT-4 nemá zásadní vliv na expresi proteinu p53, ani na expresi jeho fosforylovaných forem. Proteiny jsme kvantifikovali pomocí měření integrované optické denzity, která nám poskytla detailnější přehled o množství proteinů u jednotlivých skupin ovlivněných buněk.In the project we were focused on molecular changes activated by mitoxantrone and inhibitor U0126 (inhibitor MEK1/2) in human tumor cells. The aim of the thesis was to find if and how inhibitor U0126 is activating biochemical pathways after the DNA damage caused by mitoxantrone. As a model system we selected cell line MOLT- 4, derived from T-lymphocytic leukemia. First, we observed the dynamics of cell viability and proliferation after the effect of different doses of mitoxantrone. The object of the task was to choose suitable dose scheme for other studies. Then we observed proliferation activity of the cells after the effect of mitoxantrone and inhibitor U0126 and we evaluated the influence of inhibitor to the action of mitoxantrone. Cell line MOLT-4 after exposure of mitoxantrone and inhibition of MEK1/2 by U0126 together caused increase in dead cells. The induction of apoptosis was evaluated by flow cytometry. We used the flowcytometric methods with the application of dual mark by Annexin V and propidium iodide and optical parameters of light scattering. The induction of apoptosis was measured after the exposure of the cells to the different doses of mitoxantrone, inhibitor U0126 and their combination in the different time lags. It was proved that U0126 through the inhibition MEK1/2 stopped the pathway leading to proliferation and cell survival and after the DNA damage more cells were sent to apoptosis. For better view of the process we studied of the cell cycle, using the analysis of DNA content. In the cell line MOLT-4 mitoxantrone caused G2/M cell cycle arrest followed by apoptosis, inhibition of MEK1/2 had not any influence on the G2/M arrest. Follow-up research was concentrated on the changes in the expression of the proteins that participate in the regulation of cell cycle and progress of apoptosis (p53 and its post-translation modifications, p21, ERK1/2, lamin B). The proteins were detected by electrophoresis and Western blot followed by immunodetection. The results lead to conclusion that inhibition of MEK1/2 has not influence on the expression of p53 and its post-translation modifications in the cell line MOLT-4. Subsequently, we used the measurement of integrated optical density for the quatification of proteins. This step provided more detailed information about the amount of proteins in individual groups of affected cells.Katedra biologických a biochemických vědDokončená práce s úspěšnou obhajobo

Occurrence of Antibiotics in the Environment

5 ABSTRACT Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Marcela Seifrtová Supervisor: Prof. RNDr. Petr Solich, CSc. Prof. Angelina Pena, Ph.D. Title of Doctoral Thesis: Occurrence of antibiotics in the environment The occurrence of pharmaceuticals in the environment is a growing concern. The pharmaceuticals are developed to have biological effects, and because of their biological and physico-chemical properties, their release into the environment can have impact in human health and ecosystems. Despite their long half-life, pharmaceuticals are constantly being release into the environment, contributing to a continuous exposure of aquatic ecosystems. The pharmaceuticals are considered as emerging environmental pollutants and the assessment of their environmental risk is acquired. Research on the occurrence, fate and effects is described in the scientific literature, however the data are still insufficient. Antibiotics are an important class of pharmaceuticals widely used in human and veterinary medicine. The main problem associated with their presence in the environment is the development of mechanisms of resistance by bacteria, leading to the development of multi-resistant bacteria, which may put at risk public health in terms of..

Regulární nakrytí - struktura a složitost

Regular Coverings - Structure and Complexity Michaela Seifrtová The thesis consists of two main parts, the first concentrated on the struc- ture of graph coverings, where different properties of regular graph coverings are presented, and the second dealing with computational complexity of the covering problem. Favorable results have been achieved in this area, proving the problem is solvable in polynomial time for all graphs whose order is a prime multiple of the order of the covered graph.

A new approach for assessing the suspected pesticide poisonings of honeybees:certified methodology

Honeybee colonies are exposed to hundreds of plant protection products, which are especially due to improper use possible source of honeybee poisoning. The methodology introduces to the issue of honeybee poisoning including legislation and contributes with novel aspects to the indication and evaluation of the suspected honeybee poisonings, which are anually investigated by the State Veterinary Administration and the Central Institute for Supervising and Testing in Agriculture in Czechia. The methodology provides instructions to identify honeybee posining and notification of that finding. Moreover, possible confusions of the poisonings with honeybee diseases are highlighted. Some sections of the methodology provide innovative recommendations for improvement of honeybee poisoning assessing including determination of the extent of colony contamination. The present methodology is useful for the state administration, beekeepers, scientific and educational purposes