The dissertation is focused on the effect of three antivirals (acyclovir, rimanatadine or zidovudine) for their potencial use in the process of chemotherapy in the combination of meristem culture method as a possible method for Onion yellow stripe virus, Leek yellow stripe virus, Garlic common laten virus and Shallot latent virus eradication in in vitro conditions. Antivirals were always applied separately into cultivation media and two amounts of antivirals (25 or 50 mg.l-1) were used. The treatment time was two weeks. Meristem culture only was a control method. Murashige and Skood medium was used for all variants. During the first year, young plants of the variety D Alsace Freres cultivated in vitro were treated with chemotherapy. These plants had been treated with the meristem culture only in the past but the treatment had not been successful. The virus presence was detected with the ELISA method. Rimantadine variant in the amount of 50 mg.l-1 was the most effective variant for the monitored virus elimination. More sensitive method reverse transcription polymerase chain reaction (RT-PCR) was optimized for monitored viruses in the next parts of this work. Garlic common latent virus (GCLV) primers were designed as well. During the second year, the chemotherapy was applied on explants with the size 0.8 mm during meristem culture. Explants of the varieties Blanin, Sukoradský and Japo, however, did not regenerate and the chemotherapy could not be evaluated. In the next two years of the work, the chemotherapy was applied on individual cloves of the Unikát variety. Whilst detecting the presence of viruses in plants prior the treatment, it was established that only one plant was free from all the monitored viruses. After treatment, acyclovir variant in amount of 25 mg.l-1 was evaluated as the most effective variant for the virus complex elimination, and also from the economical point of view. Real-Time PCR method for virus detection has been started in the dissertation. The detection of GCLV was optimized from the start. Isolates which had been negative for the GCLV presence after RT-PCR were tested with Real-Time PCR. A higher percentage of GCLV positive plants were detected with Real-Time PCR. Real-Time PCR optimization for other viruses will be the objective of futur experiments.

The transplantation of bone marrow - more generally said of hematopoietic cells - is in the present time one of the modern therapeutical methods used in cases of malfunction of haematogenesis inside of the bone. The ususal reason for going through with transplantation is leukemia, lymphomas, also anemia, congenital metabolic disorders, autoimmune disorders or some of the solid tumors. Thanks to the transplantation of hematopoietic cells, it is possible to highten the chance of restoration in more than 50 % of patients suffering from oncological disease. The submitted thesis is processed in theoretical form, it is goal is to present the important points leading to progress of this presently very much used treatment and also thorough descripton of present trends in the area of transplantation of hematopoietic cells, that are almost identical in local transplatation centers as they are in the world. Another goal is to describe the role of a general nurse during providing of the care for a patient before and after transplantation of hematopoietic cells. The thesis is focused on the key areas of the issues of transplatation of hematopoietic cells and it is divided into several parts. The first part deals with the general description of the topic, it presents short introduction into the transplantation of hematopoietic cells, the basic division, historical and current indications and short preamble into todays trends. It describes closely the area of the registry of the bone marrow donors, which is the basics for the choosing of the suitable unrelated donor. The second part of the thesis deals with the desciption of historical progress since the very beginnings, through the progress during postwar period, the development of 70s and 80s of the previous century and also the 90s of the 20th century up until the present day. The third part is focused on current methods of collection of hematopoietic cells and following transplatation, it also describes closely the ways of donating hematopoietic cells, the preparation of the patient for transplantation, the course of transplantation and what should the patient prepare for during the early posttransplantation period. The fourth part is focused on nursing, it states thoroughly the role of a nurse in the nursing care before and after transplantation of hematopoietic cells, the principles a nurse has to abide by, and it also closely describes the role of a nurse during transplant application. It also describes dietary and important regimen measures the patient is set to follow after transplantation. The last part is focused especially on complications that are possible after hemopoetic cells transplantation. The necessary data have been obtained through thorough study of professional Czech and foreign monoghraphs and scientific periodicals. The acquisition of the data was significantly contributed to by the attended internsip in Internal hematological and oncological clinic of the Faculty hospital Brno Bohunice. The findings regarding nursing were obtained especially by the study of nursing standards. This thesis points out the important role of transplatation of hemopoetic cells as a method of treatment predominantly of hemooncological illnesses in current medicine and the most important roles of a nurse during the nursing of transplantation patients. The thesis provides a comperehensive overview on the issue of transplants of the hematopoietic cells and describes key areas that are considered as important. It focuses on the historical progress and the desciption of present trends thanks to which the transplantology can be considered as a highly professional and top field. It also highlights the main roles of a nurse during nursing that must be proffesional, qualified, meticulous, considerate and individualized. The results of the survey are supposed to provide the professional and amateur public enough information about the issue of transplantation of the hematopoietic cells.

Evaluation of the antiviral effectiveness of ribavirin and acyclic nucleotide phosphonates to radish mosaic virus. Virus inoculation of plants with RaMV and immunological assay of the virus by ELISA. Subsequent application of antiviral agents and monitoring relative content of the virus in plants. Subsequent processing of data in tables and graphs, and then statistical evaluation.

A new method was developed for testing antiviral compounds against plant viruses based on rapidly growing brassicas in vitro on liquid medium. While using ribavirin as a standard for comparison, phytotoxicity and ability of the acyclic nucleotide analogues(R)-PMPA, PMEA, PMEDAP, and (S)-HPMPC to eliminate ssRNA Turnip yellow mosaic virus (TYMV) were evaluated by this method. Double antibody sandwich ELISA was used for relative quantification of viral protein in plants. Ribavirin had the most powerful antiviral effect against TYMV. On the other hand, (R)-PMPA and PMEA had no antiviral effect and almost no phytotoxicity compared to the control. (S)-HPMPC and PMEDAP showed moderate antiviral effect, accompanied by higher phytotoxicity.

Acute myeloid leukemia (AML) is a very heterogeneous disease associated with cytogenetic aberrations and genetic mutations. Many of these changes have been revealed and their detection became usual part of the diagnostic process today. However, changes of expression profiles of small, noncoding RNAs, so called microRNAs (miRNAs), are less known and not used for diagnostics yet. These RNAs, 19-24 nucleotides long, take part in the regulation of expression of different genes through complementary base pairing to the 3'non- translated region (3'UTR) of the target messenger RNA (mRNA). They can influence key processes of the cell, like differentiation, proliferation or apoptosis. The changes in expression of different miRNAs are known from different types of cancers. In solid tumors, they are usually detected from bioptic samples; but also plasma samples are now in the center of attention as so called liquid biopsies providing the information about molecular genetic events in the organism. Many studies have revealed deregulated miRNAs in the bone marrow, full blood or isolated progenitor cells (CD34+) of AML patients, only four of them have analyzed plasma samples. We focused on the plasma samples and we targeted on such miRNAs, which levels differ at AML diagnosis and after the chemotherapy. Out of...

Introduction: Colorectal cancer (CRC) is one of the most frequent malignancies and affects approximately 5% of worldwide population. More than 75% of CRC cases represent sporadic forms. Susceptibility to nonhereditary CRC is significantly influenced by polymorphisms and mutations in low-penetrance genes. Variations in biotransformation and DNA repair genes may result in acumulation of toxins and DNA damage in cells leading to the development of cancer. Furthermore, different gene expression profiles of membrane transporters affecting the accumulation of anticancer drugs in tumour cells, e.g. ABC drug transporters, may largely influence inter-individual variability in drug response and chemotherapy outcome. The aim of this study was to evaluate the role of genetic and lifestyle factors in the risk of onset and progression of colorectal cancer. This study followed selected genetic alterations in xenobiotic-metabolizing enzymes (CYP1B1, GSTM1, GSTT1, GSTP1, NQO1 and EPHX1) and genes involved in response to DNA damage (CHEK2 and NBN), as potential CRC susceptibility factors. Another aim of this study was to investigate expression profile of all human ABC transporter genes to follow their prognostic and predictive potential in colorectal carcinoma. Materials and methods: The polymorphisms and other...

Chemotherapy is one of the basic therapeutic procedures of breast cancer (BC) which may precede and/or follow the surgical resection of a tumor as a part of neoadjuvant or adjuvant therapy. However, the selective pressure of chemotherapy on tumor cells may change their molecular and expression profile and thus also their chemosensitivity. The aim of our work was to document the expression changes of selected markers in BC after neoadjuvant chemotherapy, which may contribute to the understanding of the role of these proteins and genes in tumor response to chemotherapy and the development of chemoresistance. Immunohistochemical analysis of expression of standard BC markers [estrogen (ER) and progesterone receptors (PR), HER2 and proliferation activity (Ki67)] and intercellular junction proteins (claudin 1, 3 and 4, E- and N-cadherin) before and after neoadjuvant chemotherapy revealed a decrease of PR, Ki67 and claudin 3 expression and an increase of claudin 1 expression. The expression of ER, HER2, claudin 4, E- and N-cadherin proved to be stable. Assessment of standard BC markers is performed routinely during a bioptic investigation as a necessary factor for therapy indication. Our results support the current recommendations for the re-examination before indication of adjuvant chemotherapy. Claudins...

Introduction: Colorectal cancer (CRC) is one of the most frequent malignancies and affects approximately 5% of worldwide population. More than 75% of CRC cases represent sporadic forms. Susceptibility to nonhereditary CRC is significantly influenced by polymorphisms and mutations in low-penetrance genes. Variations in biotransformation and DNA repair genes may result in acumulation of toxins and DNA damage in cells leading to the development of cancer. Furthermore, different gene expression profiles of membrane transporters affecting the accumulation of anticancer drugs in tumour cells, e.g. ABC drug transporters, may largely influence inter-individual variability in drug response and chemotherapy outcome. The aim of this study was to evaluate the role of genetic and lifestyle factors in the risk of onset and progression of colorectal cancer. This study followed selected genetic alterations in xenobiotic-metabolizing enzymes (CYP1B1, GSTM1, GSTT1, GSTP1, NQO1 and EPHX1) and genes involved in response to DNA damage (CHEK2 and NBN), as potential CRC susceptibility factors. Another aim of this study was to investigate expression profile of all human ABC transporter genes to follow their prognostic and predictive potential in colorectal carcinoma. Materials and methods: The polymorphisms and other...

Introduction Axillary dissection has little diagnostic and therapeutic benefit in the node-positive breast cancer patients in whom axillary disease has been completely eradicated after neoadjuvant chemotherapy (ypN0). We sought to assess the efficacy of an algorithm used for the identification of the ypN0 patients consisting of intraoperative evaluation of sentinel and tattooed (initially positive) lymph nodes. Methods Included were T1 and T2 breast cancer patients with one to three positive axillary lymph nodes marked with carbon who were referred for neoadjuvant chemotherapy followed by a surgery. Axillary dissection was performed only in the patients with residual axillary disease after neoadjuvant chemotherapy on ultrasound or with metastases described in the sentinel or tattooed lymph nodes either intraoperatively or in the final histology. Results Out of 62 included initially node-positive patients, 15 (24%) were spared axillary dissection. The detection rate of tattooed lymph node after neoadjuvant chemotherapy was 81%. The ypN0 patients were identified with 91% sensitivity and 38% specificity using ultrasound and intraoperative assessment of both sentinel and tattooed lymph node according to the final histology. Discussion/Conclusion Lymph node marking with carbon dye is a useful and...

Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Nikola Jaščevská Supervisor: PharmDr. Anna Jirkovská, Ph.D. Title of diploma thesis: The effects of topoisomerase II beta on the sensitivity of the cancer cells to the antineoplastics Topoisomerase II (TOP II) is a cellular enzyme responsible for solving topological problems of double-stranded DNA. Alpha and beta isoforms of TOP II are different gene products having similar catalytic activities. The expression of TOP IIα is cell-cycle dependent, peaking in G2/M phase, while TOP II isoform is expressed constitutively throughout the cell cycle. It is therefore present also in non-proliferating differentiated cells. Anthracycline antibiotics are an old class of anticancer drugs, belonging to TOP II poisons. Although their clinical usefulness is high, the incidence of side effects (especially myelotoxicity and cardiotoxicity) may limit the therapy. The key role of TOP II inhibition, which is present also in cardiomyocytes, has been increasingly discussed. Dexrazoxane, the only clinically used cardioprotective, leads to depletion of TOP II in cardiomyocytes, which may explain its cardioprotection. Although TOP II was previously shown to be dispensable for cellular proliferation, its possible...