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The comparison of properties of cell lines resistant to ellipticine, doxorubicin, and cisplatin
Černá, Tereza ; Poljaková, Jitka (advisor) ; Eckschlager, Tomáš (referee)
7 Abstract Neuroblastoma is the most common extracranial solid tumor of childhood. Despite advances in cancer diagnosis and therapy, the treatment of some forms of neuroblastoma is still complicated. One of the major complications of the chemotherapy is a developed drug resistance. This master thesis deals with the effect of cytostatics on protein and gene expression of selected proteins, which may contribute to chemoresistance of the human neuroblastoma cell line UKF-NB-4. The sensitive line UKF-NB-4 and the resistant line UKF-NB-4CDDP , UKF-NB-4DOXO and UKF-NB-4ELLI were exposed to cisplatin, doxorubicin, ellipticine for 24, 48 and 72 hours. The Western blot analysis showed that cytostatic agents cisplatin, doxorubicin or ellipticine added to the sensitive neuroblastoma cell line UKF-NB-4 in amounts which are added to resistant neuroblastoma cell lines in order to maintain resistance induced expression of p53 and reduced expression of retinoblastoma protein pRb after 72 hours of cultivation. Differences in the expression of RAS protein, cytochrome P450 1A1, 3A4 and cytochrome b5 has not been shown. Changes in the expression of the studied proteins in resistant lines UKF-NB-4CDDP , UKF-NB-4DOXO and UKF-NB-4ELLI cultured with and without cytostatic agents were not detected by the Western blot analysis....
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Involvement of cellular signal systems controlled by small G-proteins in neuroregenerative processes
Dušek, Jakub ; Novotný, Jiří (advisor) ; Košek, Dalibor (referee)
G-proteins repeatedly seem as significant signal mediators according to actual "cellular signal during neuroregeneration" reviews. This thesis aimed to find out how much small G-proteins correlate with neural tissue regeneration and which molecular mechanisms induce the regeneration. The thesis mentions pathologies of nervous system which are caused by traumas or neurodegenerative diseases in wider social perspective. Pivotal part of the text describes principles of functioning for each small GTPase family and its involvement in nervous tissue repair. This thesis demonstrates crucial role of small GTPases in neuroregeneration and points to considerable therapeutic potential of these GTPases. Key words: neuroregeneration; small GTPase; small G-protein; Ras; signal systems; neurodegeneration
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The impact of Fam84b in retinal homeostasis
Raishbrook, Miles Joseph ; Procházka, Jan (advisor) ; Mašek, Jan (referee)
Fam84b is a largely unstudied protein, where its function in eukaryotic cells is unclear. This thesis work presents a FAM84B knockout mouse model and characterises the resulting retinal phenotype in detail. FAM84B KO mice were morphologically assessed by optical coherence tomography and histological processing, revealing dynamic changes stemming from the photoreceptor and pigmented epithelial layers. This potent phenotype progresses with age, spreading inwards towards the inner retinal layers, as well as laterally to adjacent retinal regions. Comparative localisation of standard retinal cell markers demonstrates that FAM84B KO retinal layering becomes increasingly disorganised, together with deformation of the retinal macrostructure. Due to this, KO mice experience reducing responses to light, as demonstrated by electroretinography, where overall retinal efficiency falls. Fam84b shows homology to the HRASLS enzyme family, which are capable of attenuating Ras-associated signalling. To investigate whether Fam84b possesses a similar function, the level of phosphorylated and activated downstream Ras effectors were compared between wild type and FAM84B KO mouse retinal lysates. A reduction of pERK1 (pY204) in KO lysates suggests that Fam84b holds some function related to this pathway downstream of Ras....
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Intenzivní chov okounka pstruhového (Micropterus salmoides) v RAS s různým světelným režimem
BOŇKO, Dominik
Largemouth bass (Micropterus salmoides) is one of the species of fish that don´t have a significant position among the species of fish breeded in Czech Republic. However the trend of building aquaculture recirculation systems with warm water comes to the fore. Breeding in these conditions opens up the possibility of introducing new kinds of thermophilic fish species. Largemouth bass definitely belongs to this group. The general factors of successful breeding of largemouth bass are the temperature and purity of the water, the suitable-composed diet and the correct stocking density. This work studies another factor that not only participates in the success of the breeding, but also has influence on the energy expenses of the aquaculture system. It is the effect of photoperiod during breeding. Five experimental groups with three repetitions were created according to the time of lighting (8, 12, 16, 20 and 24 hours of light a day). The experiment lasted 16 weeks. The average water temperature was 24,6 °C. The initial stocking density for all tanks was 20.6 kg . m-3. Fish had an initial average weight of 32.6 +- 2.1 grams. The feed ration was set up at 1% of biomass per day. As a diet were used a floating dry pellets Skretting R-2 Europa 15 F of 2 mm. The feed contained 55 % of protein and 16 % of fat. Fish were fed manually by hand. All groups showed high survival during the experiment (99.4 - 100 %). The best results were achieved by a group of 8 hours with the highest feed conversion (FCR = 0.94 +- 0.06) and the highest growth rate (SWGR = 0.6 +- 0.04 % . day-1). The average individual fish weight of the 8 hour group at the end of the experiment was 64.07 +- 2.82 g. On the other hand, group of 12 hours was the worst with the lowest average individual weight (W= 57.51 +- 1.51 g), growth rate (SWGR = 0.49 +- 0.04 % . day-1) and the lowest feed conversion (FCR = 1.16 +- 0.08). These two groups were significantly different in most production indicators. The pervisceral fat index grew with biomass. The longer the photoperiod, the higher the plasma and ammonia content of lactate. In contrast, glucose content decreased. A longer photoperiod also influenced the higher production of antioxidants.
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ADAPTACE A INTENZIVNÍ CHOV OKOUNKA PSTRUHOVÉHO (Micropterus salmoides) V POROVNÁNÍ S INTENZIVNÍM CHOVEM CANDÁTA OBECNÉHO (Sander lucioperca)
HANZLÍK, Petr
The object of this thesis is to verify Largemouth bass (Micropterus salmoides) as an alternative species to Pikeperch (Sander lucioperca) for fish farming in RAS (Recirculating Aquaculture Systems). Largemouth bass with its good meat quality is comparable to Pikeperch and with its lower sensitivity to stress related to RAS conditions could expand the number of species used in intensive aquaculture. This experiment was split to two parts. In the first part was documented adaptation of Largemouth fingerlings to RAS conditions and artificial feed. After successful adaptation, the second part was started the main experiment. Only 9-16g form pieces were picked from the fish from the first experiment.) The Fish were divided into four groups: Candát 100%, Okounek pstruhový 100%, Candát mix, Okounek mix in three repetitions to nine tanks with capacity of 600 l. Dimensions of these tanks are 101x100x59,5 cm. Each tank was settled by 700 fish. In mixed groups were 350 of Pikeperch to 350 of Largemouth bass in each tank. Second part of experiment lasted 60 days. During that time different parameters of water quality were monitored. Twice a day oxygen, water temperature and once a day pH, NO2- , NH4- were monitored. Every day feeding dosage and mortality were counted. At the end of the experiment all fish were weighed piece by piece to find out the percentage frequency of each weight category in 1 g step. From each tank after every part of the experiment 33 pc of fish were selected and the biometrics were made. Monitored parameters were TL,SL,W and FC, FCR, SGR were counted. Evaluation of outcome and comparison of researched groups pointed to next findings. Largemouth bass showed a slightly smaller growing ability than Pikeperch in RAS. We have to consider a lower water temperature to the end value 21,5°C in system which was set because of Pikeperch sensitivity to bacterial infection and the optimum water temperature for Largemouth bass to 26°C. The experiment proved a high survival of Largemouth bass in groups Okounek 100% 99,86% survival, Okounek mix 99,81% survival. Pikeperch in groups Candát 100% and Candát mix did not exceed 97% in survival. An interesting finding of this thesis proved a positive influence of Largemouth bass to the growth of pikeperch. The group Candát mix had the best results in all monitored parameters TL,SL,W and FC, FCR, SGR.
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Investigating critical mechanisms of oncogenesis using cell model systems
Hušková, Hana ; Stopka, Tomáš (advisor) ; Macůrek, Libor (referee) ; Vojtěšek, Bořivoj (referee)
(EN) Humans and cells in their bodies are exposed to various mutagens in their lifetime that cause DNA damage and mutations, which affect the biology and physiology of the target cell, and can lead to the expansion of an immortalized cell clone. Genome-wide massively parallel sequencing allows the identification of DNA mutations in the coding sequences (whole exome sequencing, WES), or even the entire genome of a tumour. Mutational signatures of individual mutagenic processes can be extracted from these data, as well as mutations in genes potentially important for cancer development ('cancer drivers', as opposed to 'passengers', which do not confer a comparative growth advantage to a cell clone). Many known mutational signatures do not yet have an attributed cause; and many known mutagens do not have an attributed signature. Similarly, it is estimated that many cancer driver genes remain to be identified. This Thesis proposes a system based on immortalization of mouse embryonic fibroblasts (MEF) upon mutagen treatment for modelling of mutational signatures and identification and testing of cancer driver genes and mutations. The signatures extracted from WES data of 25 immortalized MEF cell lines, which arose upon treatment with a variety of mutagens, showed that the assay recapitulates the...
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Physiotherapy using rhythmic auditory stimulation in upper limb training for patients with motor disorder caused by brain damage
Volchanskaya, Sofya ; Táborská, Silvie (advisor) ; Sládková, Petra (referee)
Title: Physiotherapy using rhythmic auditory stimulation in upper limb training for patients with motor disorder caused by brain damage Abstract: The theme of this bachelor thesis is physiotherapy using rhythmic auditory stimulation in upper limb training for patients with motor disorder caused by brain damage. The thesis consists from theoretical and practical parts. The range of problems associated with brain damage, both traumatic and stroke-induced, is briefly described in theoretical part. Fine and gross motor function of upper limb, the impact of central nervous system damage on it and possibilities of subsequent physiotherapy are also described in this part. In the last chapter of theoretical part knowledge about musical therapy, its application in neurorehabilitation and Rhythmical Auditory Stimulation method (RAS) is summarized. Practical part consists of two case reports of patients with acquired brain damage and following upper limb moving disorder. In this part the influence of RAS method during therapy is monitored. The evaluate the method's effect Five-Step Clinical Assessment by Gracies and Jebson-Taylor standard hand function test were used prior to initiation of treatment and immediately after its completion. The purpose of this thesis is summarizing and comparing knowledge about RAS method...
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The influence of rhythmic auditory stimulation on gait-pattern of patients with brain damage brain damage
Pavlikhina, Ekaterina ; Táborská, Silvie (advisor) ; Konopáčová, Eva (referee)
Author: Ekaterina Pavlikhina Supervisor: Mgr. Silvie Táborská Title of bachelors thesis: The influence of rhythmic auditory stimulation on gait-pattern of patients with brain damage Abstract: The Bachelor's thesis is focused on influences of musical therapy method RAS (Rhythmic Auditory Stimulation) upon the quality of patient's walking with the brain injury. The thesis summarizes and compares theoretical knowledges of music therapy in patients with pathological gait-pattern due to brain damage. Theoretical part of the Bachelor's thesis is intended to focus on the brain injury both caused by a trauma and an infarct. In the following part are described the physiologic and pathological mechanisms of walking. The final chapter of the theoretical part introduces the exact principles of musical therapy, it's variants and technics that can be used in the treatment of patients. In this chapter I have also referred on the studies that correlate the topic. The practical part contains two casuistries of the patients with the brain injury, where is described kinesiology and the therapeutic exercises in the way of the program. In order to esteem the progress of applicated therapy I used the special "walking tests". Key words: brain damage, music therapy, pathological gait-pattern, Rhythmic Auditory Stimulation, RAS
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Structure- and sequence-based identification of functionally important amino acids in a protein family
Peclinovská, Iveta ; Novotný, Marian (advisor) ; Pleskot, Roman (referee)
A group of small GTPases consist of over twenty protein families in the super class P-loop. It has a very diverse cell functions. Small GTPases regulate the formation of vesicular follicles, cytoskeleton and nuclear transport. They participate also on cell proliferation and signaling. The aim of my work is to find important amino acids that define family and distinguish each other. I focus on families Arf, Rab, Ran, Ras and Rho. At the Rho family I am also devoted to classes Rho, Rac and Cdc42. Amino acids are identified using bioinformatic programs selected Consurf and Sca5. The objective is also to test P2RANK specialized tool developed at the Charles University in Prague that predict ligand binding sites from protein structure in different families. Founding amino acids can have a big role in the functional divergence of individual families and classes of small GTPases and can be the basis for future study example for the proliferation of cancerous cells. 1.1 Keywords Powered by TCPDF (www.tcpdf.org)
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The comparison of properties of cell lines resistant to ellipticine, doxorubicin, and cisplatin
Černá, Tereza ; Poljaková, Jitka (advisor) ; Eckschlager, Tomáš (referee)
7 Abstract Neuroblastoma is the most common extracranial solid tumor of childhood. Despite advances in cancer diagnosis and therapy, the treatment of some forms of neuroblastoma is still complicated. One of the major complications of the chemotherapy is a developed drug resistance. This master thesis deals with the effect of cytostatics on protein and gene expression of selected proteins, which may contribute to chemoresistance of the human neuroblastoma cell line UKF-NB-4. The sensitive line UKF-NB-4 and the resistant line UKF-NB-4CDDP , UKF-NB-4DOXO and UKF-NB-4ELLI were exposed to cisplatin, doxorubicin, ellipticine for 24, 48 and 72 hours. The Western blot analysis showed that cytostatic agents cisplatin, doxorubicin or ellipticine added to the sensitive neuroblastoma cell line UKF-NB-4 in amounts which are added to resistant neuroblastoma cell lines in order to maintain resistance induced expression of p53 and reduced expression of retinoblastoma protein pRb after 72 hours of cultivation. Differences in the expression of RAS protein, cytochrome P450 1A1, 3A4 and cytochrome b5 has not been shown. Changes in the expression of the studied proteins in resistant lines UKF-NB-4CDDP , UKF-NB-4DOXO and UKF-NB-4ELLI cultured with and without cytostatic agents were not detected by the Western blot analysis....
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