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Genetické aspekty melanomu
KOUDELKOVÁ, Kateřina
The aim of this diploma thesis is to summarize findings about the cellular, molecular and mainly genetic characteristics of malignant melanoma, which is one of the most harmful human malignancies with steadily increasing incidence in recent decades. Malignant melanoma is a highly complex disease in which a huge number of genes play a role. The experimental part is devoted to the detection of mutations in the CDKN2A and CDK4 genes and polymorphism C609T in the NQO1 gene in participants with melanoma or pancreatic cancer in the family.
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Phenotype of melanocytes under physiological and pathological conditions
Strnadová, Karolína ; Lacina, Lukáš (advisor) ; Mokrý, Jaroslav (referee) ; Balvan, Jan (referee)
In addition to the dominant keratinocytes and fibroblasts, melanocytes are also indispensable representatives of skin cell populations. Melanocytes are pigment cells whose primary function is to produce the pigment melanin, which is important for protecting keratinocytes from harmful ultraviolet radiation. Excessive exposure to this radiation is a risk factor for the development of skin tumours, including malignant melanoma of the skin, in which pathological transformation of melanocytes into melanoma cells occurs. The presented thesis focuses on 4 thematic areas associated mainly with malignant melanoma. In the first thematic area, the increasing incidence of malignant skin melanoma is associated with the ageing of the population. One of the reasons seems to be the more frequent occurrence of proinflammatory setting in the ageing organism. It prepares a suitable environment for tumour development. The second thematic area focuses on new approaches that could expand the range of diagnostic methods for the early detection of malignant melanoma. The first approach methodically uses the detection of proinflammatory molecules in the patient's serum. Higher serum levels of IL-6 and IL-8 correlate with an unfavourable patient prognosis. The second approach is based on the possibility of detecting a...
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Studies of intercellular interactions in tumours
Jechová, Alžběta ; Smetana, Karel (advisor) ; Skalníková, Helena (referee) ; Masařík, Michal (referee)
Beside tumor cells themselves, tumors consist of many non-malignantly transformed cellular elements and an extracellular matrix. This so-called tumor microenvironment, or stroma, significantly influences the biological properties of the tumor through intercellular interactions. In this thesis I have focused on the study of tumor-associated fibroblasts in squamous cell carcinomas of the head and neck, malignant melanoma and glioblastoma. The data show the presence of cells with mesenchymal characteristics, present even in the glioblastoma stroma, which could potentially have a positive effect on proliferative activity and invasiveness of glioblastoma cells. In malignant melanoma, the presence of keratinocytes should also be considered, as they are the major cells of the epidermis influencing tumor melanocytes. The conditioned medium from UVB irradiated keratinocytes and non-irradiated fibroblasts stimulates the invasion of malignant melanoma cells. Targeting the tumor stroma may be a new direction in oncological therapy, so we have focused on the influence of synthetic polyamine on the formation of myofibroblasts, which are an active part of the population of tumor-associated fibroblasts. The tested polyamine prevents the formation of myofibroblasts but has no effect on those already formed nor on...
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Metabolic Effects Of Hyperthermic Isolated Limb Perfusion (HILP) in Malignant Melanoma Patients
Hodková, Gabriela ; Semrád, Michal (advisor) ; Štádler, Petr (referee)
Hodková, Gabriela - Metabolic Effects Of Hyperthermic Isolated Limb Perfusion (HILP) in Malignant Melanoma Patients First Faculty of Medicine, Charles University in Prague, Praha 2, Kateřinská 32 Head of the work: Doc. MUDr. Michal Semrád, CSc. Supervisor - consultant: MUDr. Miroslav Špaček, Ph. D. The aim of the study is to assess the metabolic consequences of mechanical isolation and hyperthermic cytostatic perfusion in a limb affected by malignant process. The theoretical part refers to a topic of malignant melanoma, its clinical evaluation and treatment. Methods based on conservative and surgical treatment are described. The isolated hyperthermic cytostatic limb perfusion is a consecutive local treatment indicated in cases of recurrent malignant lesions following surgical resection, when next surgery is impossible. In the practical part, the laboratory samples and clinical data were recorded in patients who had undergone hyperthermic cytostatic limb perfusion in the 2nd Surgical Department of The General Teaching Hospital and First Faculty of Medicine, Charles University Prague. The affected limb was flushed with a warm oxygenated blood containing cytostatic drugs using an extracorporeal circuit apparatus. Selected arterial blood gas, metabolic and hematologic parameters were studied intra and...
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Studies of intercellular interactions in tumours
Jechová, Alžběta ; Smetana, Karel (advisor) ; Skalníková, Helena (referee) ; Masařík, Michal (referee)
Beside tumor cells themselves, tumors consist of many non-malignantly transformed cellular elements and an extracellular matrix. This so-called tumor microenvironment, or stroma, significantly influences the biological properties of the tumor through intercellular interactions. In this thesis I have focused on the study of tumor-associated fibroblasts in squamous cell carcinomas of the head and neck, malignant melanoma and glioblastoma. The data show the presence of cells with mesenchymal characteristics, present even in the glioblastoma stroma, which could potentially have a positive effect on proliferative activity and invasiveness of glioblastoma cells. In malignant melanoma, the presence of keratinocytes should also be considered, as they are the major cells of the epidermis influencing tumor melanocytes. The conditioned medium from UVB irradiated keratinocytes and non-irradiated fibroblasts stimulates the invasion of malignant melanoma cells. Targeting the tumor stroma may be a new direction in oncological therapy, so we have focused on the influence of synthetic polyamine on the formation of myofibroblasts, which are an active part of the population of tumor-associated fibroblasts. The tested polyamine prevents the formation of myofibroblasts but has no effect on those already formed nor on...
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Detection of malignant melanoma in histological sample using deep neural networks
Frey, Adam ; Lokoč, Jakub (advisor) ; Straka, Milan (referee)
The aim of this thesis is to create a classification method for detection of ma- lignant melanoma in high-resolution digital images. Deep convolutional neural networks were used for this task. At first, a short overview of malignant melanoma and ways to detect it is presented. Deep convolutional neural networks are also introduced with a special attention given to models used further in this work. Several ways to generate samples from the provided histological images are discussed, and several experiments are evaluated to decide how to maximize the accuracy of employed classification methods. The thesis then focuses on several neural network structures used for image classification and their possible utiliza- tion for the given task. The emphasis is laid on the transfer learning, a method used for modifying already trained models for different tasks. This method is then used for training several classifiers. Further on, several methods for the visualization of model results are discussed with some of them implemented. The experiments show promising results on par with other studies dealing with similar problems. Several possibilities for further development are listed in the conclusion.
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Construction of mouse polyomavirus chimeric VLP bearing melanoma epitopes
Kojzarová, Martina ; Drda Morávková, Alena (advisor) ; Tachezy, Ruth (referee)
Major capside protein of Polyomaviridae family viruses is able to selfassemble into virus-like particle (VLP) even without the presence of minor proteins, bind exogenous DNA non-specifically and recognise the receptor on the cellular surface. These characteristics determine its use as vector in gene therapy or immunotherapy. It was discovered before that MPyV VLPs significantly stimulate immune system and have strong adjuvant effect. Chimeric VLP derived from mouse polyomavirus carrying exogenous antigene or epitop is supposed to elicit specifically targeted immune response after immunisation. The main obstacle is choice of immunogene that is strong enough to cause adequate immune response. The goal of this thesis was to construct chimeric particles carrying epitop of malignant melanoma, one of the most immunogenic tumours, on their surface, using methods of genetic engineering. For future research of particle's immunogenic properties three types of particles were developed - particles with human and mouse melanoma epitopes, respectively and control particles with ovalbumine epitop. For the purpose of production of chimeric protein was used baculovirus expression system. It was verified then, with the use of electron microscopy, that introduction of tumour antigen into one of surface loops of VP1...
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Metabolic Effects Of Hyperthermic Isolated Limb Perfusion (HILP) in Malignant Melanoma Patients
Hodková, Gabriela ; Semrád, Michal (advisor) ; Štádler, Petr (referee)
Hodková, Gabriela - Metabolic Effects Of Hyperthermic Isolated Limb Perfusion (HILP) in Malignant Melanoma Patients First Faculty of Medicine, Charles University in Prague, Praha 2, Kateřinská 32 Head of the work: Doc. MUDr. Michal Semrád, CSc. Supervisor - consultant: MUDr. Miroslav Špaček, Ph. D. The aim of the study is to assess the metabolic consequences of mechanical isolation and hyperthermic cytostatic perfusion in a limb affected by malignant process. The theoretical part refers to a topic of malignant melanoma, its clinical evaluation and treatment. Methods based on conservative and surgical treatment are described. The isolated hyperthermic cytostatic limb perfusion is a consecutive local treatment indicated in cases of recurrent malignant lesions following surgical resection, when next surgery is impossible. In the practical part, the laboratory samples and clinical data were recorded in patients who had undergone hyperthermic cytostatic limb perfusion in the 2nd Surgical Department of The General Teaching Hospital and First Faculty of Medicine, Charles University Prague. The affected limb was flushed with a warm oxygenated blood containing cytostatic drugs using an extracorporeal circuit apparatus. Selected arterial blood gas, metabolic and hematologic parameters were studied intra and...
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Search for new biomarkers for screening and early diagnosis of cancer
KALČÍKOVÁ, Kateřina
Thesis on the topic "Search of New biomakers suitable for screening and early diagnostics of cancer" is dedicated to the issue of cancer, a cancer of the breast, prostate, ovarian and after the new findings out as melanom determination of tumor markers and their implications for clinical practice. Their aim is skouted up biomaker suitable for screening and early diagnostics of carcinoma( cancer ) of the breast, ovarian, prostate and malignant melanoma with utilization of various kinds of immunoassay. The theoretical part gives a brief and comprehensive overview of the epidemiology of selected tumors and tumor markers used and their qualities. In the methodological part are given principles applied iminoanalytics methods a list of the statistical methods and a detailed description of the groups of patients. Results section contains tables with results of biomarkers diveded by sort of avalueted tumors. In the discussion are then analyzed facts that follow from the results. It discussed the process and actualy status of searching convenients biomekers and their combinations that can distinguish a population of benign tumors of the population with malignant tumors and markers which are able to distinguish healthy population from the population with malignant tumors of the breast, ovary, prostate and melanoma. This work may serve as a source of knowledge and results to continue in search of such marker that could be used for screening and early diagnosis of cancer and so to improve the prevention of these illneses in the population.
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Use of Imunohistochemical Methods in Diagnostics of Neoplasms
VELKOVÁ, Michala
This thesis deals with the application of immunohistochemical methods in the diagnosis of neoplasms. Indications of immunohistochemical methods of particular departments in Hospital Písek, a.s. are outlined there. It describes the origins and development of this method in the Pathological Anatomy Department, Hospital Písek, a.s. In the theoretical part, I describe knowledge of single methods. As for immunohistochemistry, I focused on the determination of antigens and the use of suitable primary antibodies for diagnosing diseases, mostly neoplasms. There are 20 primary antibodies available at the Department of Pathological Anatomy. In the practical part, I compare conventional staining and immunohistochemical methods with conventional staining and special staining methods in histology for the diagnosis of tumours. Hematoxylin eosin serves for distinguishing of essential cellular components. Selective staining enables colour differentiation of various tissue components. Using immunohistochemistry, the presence of certain tissue antigens by antibody reaction with the immobilized chemical compounds is detected. This comparison I was applied at the differential diagnosis of malignant melanoma and myogenic tumours at the Pathological Anatomy Department, Hospital Písek, a.s. For the determination of malignant melanomas, initially the sample was histologically processed and stained with Hematoxylin eosin staining base. Secondarily, the argentaffin reaction was used to detect melanin pigment and immunohistochemical staining of antigen HMB 45 using a mouse monoclonal antibody against HMB 45 by Dako Company and S 100 protein using a polyclonal rabbit antibody against S 100 by Dako Company. 14 samples of patients with malignant melanoma were examined this way. HMB 45 was stained positively in 12 samples out of 14 in total. Monoclonal antibody against HMB 45 is for malignant melanoma quite specific. S 100 protein was detected in all tissue samples. Polyclonal antibody against S 100 shows higher sensitivity, however it is not very specific. It is more used to distinguish neuroectodermal and nonneuroektodermálních lesions. Melanin was detected in 11 tissue samples. Detection of melanin is also non-specific and complicated. Argentaffin reaction shows colouring of other substances contained in the cytoplasm, such as lipofuscin therefore it is necessary to use other methods for the melanoma diagnosis, e.g. immunohistochemistry. In the diagnosis of myogenic neoplasms, Hematoxylin eosin basic staining was used and subsequently immunohistochemical examination followed to detect smooth muscle actin and desmin using monoclonal mouse antibodies by Dako Company and transparent Hematoxylin van Gieson staining, which is used less than in the past. A total of 94 myogenic neoplasms (leiomyoms uterus) were examined. 72 neoplasms stained with Hematoxylin van Gieson showed a positive colour reaction of muscle mass and 22 neoplasms weak colour reaction. Smooth muscle actin and Desmin was detected by immunohistochemistry in 92 samples. The main problem of Hematoxylin van Gieson staining remains less specificity and sometimes ambiguous differentiability of colour tones in their own preparation impeding and preventing to set a reliable diagnosis in some cases. For this reason, immunohistochemical detection of Smooth muscle actin and Desmin (amongst others) is preferred as key parameters. Preferably, you can then use positive antibody reaction with Smooth muscle actinof myoepiteliels in the search for possible invasiveness mainly to differentiate in situ and microinvasive tumous in glandular organs. The differential diagnosis of myogenic origin tumours currently prefers immunohistochemical methods to special staining for high sensitivity and specificity, as well as in the differential diagnosis of malignant melanoma. When combined, both approaches complement each other well and provide relatively high degree of certainty in determining cells origin.
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