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Polyethylene wear particles around joint replacements - their properties, distribution and possible mechanism of their adverse biological effects
Zolotarevová, Eva
This PhD thesis in essence rounds off the range of queries asked several years ago when the task was set by Doc. RNDr. Zdenek Horak, PhD. The original isolation procedure for polyethylene wear particles, identification of some of their physico-chemical properties and manipulation accordingly has been successful in the process of this thesis accomplishment. It became possible to quantify the particles at relative as well as absolute scale. The thesis managed to disprove the published opinion that centrifugation can affect the morphology of isolated particles. The issue of distribution of polyethylene wear particles around hip endoprothesis has been solved at two levels: "macroscopically", the distribution of wear particles in zones around endoprosthesis was showed (this thesis does not include these data). Furthermore, at the "microscopic" level, the distribution of wear particles in granuloma tissue from the zones surrounding endoprostheses with most particles has been determined. The granuloma tissue formed in zones with most particles because of that. It was observed that the common term - osteoagressive granuloma - used by orthopedist is rather misguided. Granuloma is not aggressive to bone tissue. On the contrary, the granuloma formation reduces the effective amount of wear particles with...
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Polyethylene wear particles around joint replacements - their properties, distribution and possible mechanism of their adverse biological effects
Zolotarevová, Eva ; Gallo, Jiří (advisor) ; Rauch, Pavel (referee) ; Horák, Zdeněk (referee)
CHARLES UNIVERSITY IN PRAGUE FACULTY OF SCIENCE DEPARTMENT OF BIOCHEMISTRY PRAGUE 2010 Supervisor: Doc. MUDr. Jiří Gallo, PhD. Supervisor-consultant: Prof. RNDr. Gustav Entlicher, CSc. Eva Zolotarevová Summary of PhD Thesis POLYETHYLENE WEAR PARTICLES AROUND TOTAL JOINT REPLACEMENTS - THEIR PROPERTIES, DISTRIBUTION AND POSSIBLE MECHANISM OF THEIR ADVERSE BIOLOGICAL EFFECTS PRAGUE 2010 The research project was supported by the Grant project of National research program II. (no. 2B06096) of the Czech Ministry of Education, Youth and Sports named "Observation and minimization of UHMWPE wear in joint replacements". Front-page picture: Polyethylene wear particle isolated from granuloma tissue surrounding total hip replacement; Electron microscope microphotograph; particle size app. 90 x 80 µm; retouched Eva Zolotarevová Introduction Introduction For many people all over the world a replacement with an implant of irreversibly damaged joint is often the only way how to come back to normal life without pain and mobility limitation. Nowadays, greater and greater requirements are asked of quality of all replacement components, especially in terms of quality of used materials. Biomechanical properties of hip and knee replacements have been already solved. The most often used materials for manufacturing joint...
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Analýza variability genomů Nepovirů (GFLV a ArMV) v produkčních vinicích vinařské oblasti Morava
Eichmeier, Aleš
The disertation thesis was focused on the analysis of variability of two nepoviruses genomes, Grapevine fanleaf virus -- GFLV and Arabis mosaic virus -- ArMV. The thesis described single isolates of these nepoviruses and their until recently non-explored genome portions coded by RNA1 strand, the coding regions for 1BHel and 1EPol proteins precisely. The main topic of this thesis was the identification of infected plants that were infected by nepoviruses GFLV and ArMV. Based on design of specific primers there was established sequence homology of coding sequences. Another topic was the development and optimalization of diagnostic system for the efficient GFLV and ArMV detection. This work was based on hypothesis of very high level GFLV and ArMV genomes variability. The results were recieved mainly from sequencing analyses performed by capillary automatic sequencer ABI-PRISM 310 (Applied Biosystems, Carlsbad, USA). Obtained results were assessed with CLC Main Workbench 5 software (CLC Bio, Aarhus, Denmark). Real-time PCR TaqMan multiplex was also designed using this software. This real-time tool is able to detect and quantify both nepoviruses within one PCR reaction simultaneously. There were sequenced and analysed the genetic codes at the nucleotide level for MP 2B , 2CCP, 1BHel and 1EPol proteins in this work. The first one was assessed because of the development of real-time PCR TaqMan multiplex. In this case were sequenced 290 bp from 14 GFLV and ArMV isolates, the variability level was established in frame of nucleotides 71.7-97.6%. The partial genetic code for 2CCP was assessed in frame of 6 GFLV isolates and the variability was established in 935 bp fragment to 83-86% at the nucleotide level and 81-91% at the amino acid level. The partial genetic code for 1BHel was sequenced in frame of 6 isolates GFLV and the variability was established in 379 bp in frame of all accessible GFLV and ArMV isolates, the variability was established 86.4-98.9% at the nucleotide level and 96-100% at the aminoacid level. The partial genetic code for 1EPol protein was sequenced in 6 GFLV isolates in frame of 365 bp and the variability was established for all accessible sequences in GenBank/NCBI. The variability was established at the nucleotide level 79.5-100% and at the aminoacid level 91.4-100%. All received sequences longer than 300 bp were submitted to GenBank/NCBI. The obtained partial genetic codes for 1BHel a 1EPol proteins coded in RNA1 were unique at the time of publication. This work provided the important results which were used subsequently like source information for construction of vectors intended for the preparation of transgenic plants. Then the incurred grapevine would show resistance against GFLV. Moreover, based on results of this work there was created certified methodology of the real-time PCR TaqMan multiplex method which can be standartly used as an efficient diagnostic tool for simultaneous detection of GFLV and ArMV. This real-time system was succesfully compared with ELISA what would be interesting mainly for diagnostic laboratories engaged in the certification processes of propagation material.
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Non-contrast MRI perfusion imaging
Kruml, Ondřej ; Jiřík, Radovan (referee) ; Krátká, Lucie (advisor)
This bachelor paper is focused on the magnetic resonance perfusion imaging technic without the usage of an external contrast media. This paper contains a look at the basic functional principle of the magnetic resonance. The focus is then set set on the Arterial Spin Labeling (ASL) technic. The basic principle of ASL uses protons from the water mocelules in arterial blood as an endogenous contrast agent. This technic can be used for obtaining a perfusion map of any tissue without the usage of invasiont echnics or contrast agents. The quantification of perusion was done applying the FAIR-RARE scheme, which belongs to the large family pulsed ASL.
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Classification of marked cells migration in tissue
Solař, Jan ; Skopalík, Josef (referee) ; Čmiel, Vratislav (advisor)
This diploma thesis deals with analysing of modern methods for cell detection, visualization and quantification in 3D space. The first section deals with optical methods for cells detection. There is detailed discussion about cell labeling and detection on confocal microscopy. There is also description about developed algorithm for whole cell volume quantification from microscopy images. This could made a comparsion of fluorescence signal according to time of cell labeling and according to cell shapes. There was also optimalization of handmade tissue phantoms visualization. It could be compared a possibilities of cell detections in these phantoms by confocal microscopy and OCT. It was also implemented algorithm for quantification of cells from OCT images. Besides confocal microscopy and OCT cells are also analyzed by other methods. The last part is the Conclusion of results and comparison of used methods.
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