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Účinky xenobiotik na oxidační stres, metabolizmus lipidů, integritu DNA a životaschopnost lidských buněk a rybích spermií in vitro
LINHARTOVÁ, Pavla
Pollution of the aquatic environment by inorganic and organic chemicals is a major factor posing a serious threat to the survival of aquatic organisms including fish. In addition balancing risks and benefits of fish consumption is nowadays an intensively discussed public health topic. Spermatozoa of almost all fish species are released into water environment where they can be directly exposed to various compounds, such as xenobiotics including toxic metals, prior to fertilization. In addition, exposure of parental adults to various xenobiotics may affect gamete quality, which may subsequently reduce fertilization success. On the other hand the advantages of eating fish are well-known, not only in the point that fish is a healthy source of protein and other nutrients, but eating contaminated fish may also confer various health benefits. Research over the past few decades has shown that the nutrients and particularly the n-3 fatty acids (FA) found in fish and seafood, are for examples protective against cardiac diseases and have a positive impact on brain development. The thesis provides a focus on two different cell model types. Firstly, human hepatocellular cells (Hep G2, ATCC) were used as in vitro tool for studying the effect of the intake of cadmium (Cd2+) contaminated fish on cytotoxicity, oxidative stress and fatty acid and phospholipid class compositions. Secondly, spermatozoa of one threatened species of fish, sterlet (Acipenser ruthenus) were used as in vitro model for studying effect of potentially hazardous xenobiotic compounds' occurring in open waters. Sperm from sterlet were exposed for 2h to environmentally relevant concentrations of DQ (0-150
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Alakali-metal-cation homeostasis in pathogenic Candida species
Elicharová, Hana ; Sychrová, Hana (advisor) ; Heidingsfeld, Olga (referee) ; Půta, František (referee)
Several tens of Candida species belong to the opportunistic human pathogens capable of inducing life-threatening infections in immunocompromised patients. Virulence of single Candida species depends among others on their resistance to the variable external conditions. The maintenance of alkali-metal-cation homeostasis, which means the ability to accumulate sufficient amount of potassium cations and on the other hand to survive under high extracellular concentrations of alkali-metal cations, is essential for growth and virulence of Candida cells. We observed the negative effect of fluconazole (FLC) on salt-tolerance of six Candida species and found that it is independent of the species level of FLC- resistance. FLC hyperpolarizes plasma membrane of Candida cells and therefore increases non-specific uptake of alkali-metal cations which results in strongly increased salt-sensitivity of Candida cells. The FLC-induced hyperpolarization also results in an increased sensitivity of Candida cells to the antifungals which are positively charged and are driven into the cells by the membrane potential. The effect of fluconazole on membrane potential and thus on the uptake of alkali- metal cations into the cell turned our attention to the homeostasis of potassium cations whose high intracellular concentration is...
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Fanconi anemia and pancreatic cancer
Hucl, Tomáš ; Špičák, Julius (advisor) ; Nečas, Emanuel (referee) ; Vodička, Pavel (referee)
Inactivation of the Fanconi anemia (FA) pathway occurs in diverse human tumors including pancreatic cancer and renders those tumors hypersensitive to DNA interstrand-cross-linking agents (ICL). How to treat specificly pancreatic and other cancers harboring FA mutations has recently raised great interest, yet preclinical studies have been hampered by the lack of well-controlled human cancer models. We endogenously disrupted FANCC and FANCG in an adenokarcinoma cell line and observed a typical phenotype of FA pathway deficiency (abrogation of FANCD2 monoubiquitination; chromosomal instability, G2M arrest and decreased proliferation upon treatement with ICL, spontaneous chromosomal breakage). Homozygous deletion was achieved for FANCC and FANCG but not for FANCD2 and BRCA2/FANCD1 in RKO cells, suggesting a detrimental phenotype. It provided direct evidence for the paradoxical assumption that their inactivation could be predominantly selected against in cancer cells. Using high-throughput screening, we assessed the growth of our isogenic FANCC and FANCG cells upon treatment with 880 active drugs and 40 000 diverse compounds. The compound having the stronges effect, named 80136342, had a distinct mechanism of action from that of ICL agents. When applied in combination with ICL agents, 80136342 had at least...
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Variability of eggshell colouration
Bubeníčková, Kateřina ; Svobodová, Jana (advisor) ; Sládeček, Martin (referee)
Bird´s Eggs Are Specific for Big Variability in Eggshell Coloration. This Eggshell Eoloration Is Caused by Two Pigments; Biliverdin and Protoporphyrin. Biliverdin Causes Bluegreen Color of the Egg and Protoporphyrin Causes RedBrown and Black Maculas on the Surface of the Eggshell. Many Hypotheses Try to Clarify the Reason of Resultant Variability in the Eggshell Coloration. The Final Reason of the Variability in the Eggshell Coloration Has Not Been yet Explained and this Is the Reason, Why this Topic Is Subject of Scientifical Researches. In the Theoretical Part of this Thesis Are Described Individual Hypotheses, Which Describe Variability in the Eggshell Coloration. These Theses Were Focused on Structural and Signal Function of Pigment and the Anemic Hypothesis; these are Explained in Detail in the thesis, because they are the subjects of this thesis´s Research. Signal Hypothesis Works with Theory, That Egg´s Pigmentation Informs the Partner about Female´s Condition Quality. Results of Recent Published Studies, Which Are Focused on Signal Function, Say, That 60% of Results Support the Theory, That Egg Maculation Relates to Female´s Body Condition. 80% of Studies about Signal Function Made There Researches on Passerines. This Is the Main Reason, Why Signal Function of Pigment Cannot Be Applied Generaly on All Bird Species, because the Signal function Was Not Properaly or None Tested on Other Species Than Passerines (Out of Few Exceptions). Anemic Hypothesis Might Explain the Variability in the Eggshell Coloration with Theory, Protoporphyrin, Which Is Derived in Higher Concetration from Blood Due the Anaemia Causes Higher Maculation on the Surface of Eggshell. Purpose of Practical Part of this Thesis Was Finding Out, Whether Female´s Condition Quality Depends on the Character of Color Maculation in Great tit (Parus major). Futhermore if There Exist Correlation between Ratio (of Immature Red Blood Cells and Total Sum of Red Blood Cells) and Eggshell Coloration. Results of the Study Show, that the Eggshell Coloration May Depend on female´s Conditon. Specifically, Red-Brown Macules with Lower Level of Intensity and Saturation Pronounce Larger Area of Melanin Based Ventral Stripe. In Conclusion, this Thesis Supports Signal Function Thesis of Protoporhyrin and Leads to the Fact, that Final Eggshell Coloration Has an Impact on the Female´s Body Condition. Results of this Thesis Does Not Prove any Relationship Between Ratio (of Immature Red Blood Cells and Total Sum of Red Blood Cells) and Eggshell Coloration. It Leads to the Fact, that there Is No Possible Support for Correlation between Anaemia and Variability in Eggshell Coloration in our Population of Great tit.
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The Synthesis and Characterisation of Biocomposite Materials for Potential Application in Medicine
Balgová, Zuzana ; Plešingerová,, Beatrice (referee) ; Bakoš, Dušan (referee) ; Palou, Martin (advisor)
Dizertační práce se zabývala syntézou a studiem kompozitních materiálů pro potenciální lékařské využití. Teoretická část je zaměřena na biomateriály, zejména na kompozity složené z polyvinylalkoholu a hydroxyapatitu(PVA/HA). Byly připraveny kompozitní membrány složené z polyvinylalkoholu s různým hmotnostním zastoupením hydroxyapatitu - 0%, 10%, 20%, 30%, 40% a 50%. Hydroxyapatit (HA) byl připraven srážecí metodou z hydrogenfosforečnanu amonného a tetrahydrátu dusičnanu vápenatého ve vodném alkalickém prostředí. Vzniklá suspenze se smísila s roztokem polyvinylalkoholu, který byl připraven rozpuštěním ve vodě o teplotě 85° C. Jednotlivé směsi byly odlity do formy a sušeny po dobu 7 dní při teplotě 30 ° C, vzniklé 0,5 mm tenké membrány byly analyzovány ATR-FTIR spektroskopií k identifikaci funkčních skupin v kompozitu, dále byla provedena XRD analýza. Zkouška tahem a TGA měření byly realizovány k určení vlivu HA na mechanické vlastnosti, respektive změnu tepelné odolnosti kompozitů ve srovnání s čistým PVA. Byla provedena zkouška bioaktivity v simulovaném krevním roztoku (SBF) po dobu 2h, 7 a 28 dnů. SEM byla použita k charakterizaci povrchové mikrostruktury biocompositních membrán před a po ponoření do SBF. Na povrchu testovaných membrán vznikla vrstva apatitu, která je charakteristická pro bioaktivní materiály. Bylo zjištěno, že s rostoucím množstvím HA částic docházelo ke vzniku aglomerátů v kompozitu, které vznikly mimo jiné jako důsledek růstu krystalů HA během sušení membrán. Bioaktivita rostla s delším působením SBF na vzorky.
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Assisted reproduction in dogs
Kopecká, Iveta ; Tůmová, Lenka (advisor) ; Šimoník, Ondřej (referee)
The dog is among the largest and longest man domesticated beast and holds in his life indispensable place. To preserve the species is a precondition for its ability to reproduce. To increase the success rate of reproduction contribute in recent decades and methods of assisted reproduction. Among the most common methods of assisted reproduction in dogs include artificial insemination. This can be done in several ways depending on the site deposit seed in genital tract of female (intravaginal, transcervical intrauterine, intratubular insemination). It can be carried out using native, chilled or frozen semen. Insemination process itself affects the success of pregnancy rates. Generally, that is the most successful design insemination with fresh semen intratubular. Another method is assisted reproduction in vitro oocyte fertilization and subsequent embryo transfer. The success of this method is dependent on many factors, among which include the sampling of gametes method of storage, the composition of culture media, mechanisms regulating nuclear oocyte maturation and others. It is evident that the need of further research, leading to the understanding of the molecular mechanisms that control oocyte maturation and embryonic development in dogs. A relatively new method of reproductive biotechnology is somatic cell nuclear transfer into an oocyte depleted own genetic information called cloning. This technology does not achieve major success so far. Methods of assisted reproduction in dogs hinders overall physiology of the reproductive system, but also more difficult method of obtaining oocytes and embryos, and then find the appropriate recipient. The key seems to be to find a more suitable solution for long term storage of gametes. Achieving successful freezing of sperm and oocyte cryopreservation would lead to greater use of reproductive technologies. Studies on assisted reproduction in dogs are among other things contributing to overall understand of reproductive mechanisms. Acquired knowledge may enable the conservation of valuable dog genotypes in vitro.
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Hanacky estate - construction and technological project
Troubilová, Hana ; Kotek, Lukáš (referee) ; Biely, Boris (advisor)
In this diploma thesis is designed replica Hanacky estate from the 19th century. The whole structure includes not only a replica of the estate but also including the reconstruction of a barn or shelter. Hanacky estate will be part of Zoopark Vyškov and will serve as an environmental education center. Thesis I deal with a construction technological project. I focus here on the transport links, building-site cell site equipment, calculation of water and electricity, design tools and machinery, calculation, budget and schedule, the use of machines and workers, limitky. I deal with the technological regulations innovative technology. Furthermore, it is the control and test plans, security, ecology, design contract for work. In the last part of my thesis I deal with natural building materials.
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The study of the role of cell populations in rejection and tolerance of orthotopic corneal transplants
Sedláková, Klára ; Filipec, Martin (advisor) ; Jirásková, Naďa (referee) ; Čejková, Jitka (referee)
Small animal models of orthotopic corneal transplantation offer many advantages for the study of immune mechanisms after grafting - not only because of the similar mechanisms of murine and human corneal transplant rejection but also due to the feasibility of the direct observation of the animal without the need to sacrifice it. The purpose of the thesis was to study this model in mouse and rat. We established allotransplantation (BALB/c to C57BL/6 mouse and Wistar Furth to Lewis rat) and concordant xenotransplantation model (rat to mouse; Lewis to BALB/c and Sprague Dawley to BALB/c) and set up grading schemes for the evaluation of the clinical course after grafting. Initially, we focused on the effect of the suturing technique on the survival of xenografts and on the efficacy of selected immunosuppressants: cyclosporine A, monoclonal antibody against T cells (anti-Thy-1.2) and AMT (a specific inhibitor of inducible nitric oxide synthase, 2- amino-5.6-dihydro-6-methyl-4-H-1.3-thiazine)). The results demonstrate that the suturing technique significantly affects the outcome of transplantation and, importantly, influences the effectiveness of immunosuppressive regimens and therefore must be taken into account when evaluating the efficacy of immunosuppressive drugs. FTY720 is a novel immunosuppressant with a...
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Sources and Importance of Natural Antioxidants for Human Health
Frühaufová, Lenka ; Rambousková, Jolana (advisor)
Existence of free radicals as participants in biological processes, scientists expected at the beginning of the 20th century . Given the characteristics of these substances, their instability and very short half-life , however, was their exploring challenging to impossible , and therefore a deeper learning to occur with the development of relevant research methods. Hand in hand with uncovering secrets and patterns of oxidative damage to biological macromolecules occurred to focus attention also to a system that is capable of this action prevent and maintain a balance in the cell , the antioxidant system . It is formed diverse group of substances of enzymes catalyzing redox reactions through plasma proteins to substances that the human body is not capable of self- produce and must be obtained from food . The subject of the research are now interaction of both systems and applications, primarily acquired knowledge into practice. What is the role of free radicals in the pathogenesis of disease states such as cardiovascular disease or malignancy ? How to participate in prenatal damage to the fetus ? They are involved in the aging process ? Together with obtaining answers to these and many other questions are logically appears another : we can use substances having antioxidant effects in preventing or treating...
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