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Detection of human adenoviruses from the respiratory samples from the patients of Motol University Hospital
NOVÁKOVÁ, Veronika
Human adenoviruses are world-wide pathogens causing endemic and epidemic outbre-aks of disease. Adenoviral infections are related with high morbidity and mortality especially among the paediatric allogeneic haematopoietic stem cell recipients. Presented work is comparing the commercial and in-house assay for detection of the adenoviruses to improve the diagnostics of these viruses in Motol University Hospital, especially among the immunocompromissed patients, in which is the rapid and precise detection highly important. Theoretical part of work presents the characteristics of adenoviruses, clinical symptoms and diseases of the adenoviral infection, their use in the experimental therapy. Practical part is aiming the screening of the respiratory tract samples with quantitative in-house test and comparing of the results with previously performed detection by commercial Anyplex TM II RV16 test. Detection of adenoviruses was performed in 1,881 samples from 1,420 patients with symptoms of respiratory tract infection by real-time PCR. In total, adenovirus was detected in 187 samples (9.9%) from 169 patients (11.9%). One hundred samples (53.5%) was positive by both methods, 26 (13.9%) with commercial detection only and 61 (32.6%) was positive only with in-house assay. Most frequently, adenoviruses were detected among the patients from Dept. of Paediatrics, Dept. of Paediatric Haematology and Oncology and from adult patients from 3rd Dept. of Surgery and Dept. of Anaestesiology, resuscitation and intensive medicine. In all positive samples, subsequent sequence analysis of hypervariable part of 1-6 hexon gene was performed and representative sequence for identification was obtained in 70 samples. Most frequently detected adenovirus genotypes were C2 (25.7 %), A31 (24.3 %), B3 (15.7 %) a C1 (11.4 %). From 26 samples positive by Anyplex TM II RV16, only 3 samples were identified (all as genotype B3). Contrary, among 61 positive samples detected only with in-house assay, 15 samples were identified as genotype A31 and 3 as AdV from group C. Our results shows the lower sensitivity of the commercial test in detection of adenoviru-ses.
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The role of the Smc5/6 complex in DNA viral infection
Protivová, Eliška ; Huerfano Meneses, Sandra (advisor) ; Pokorná, Karolína (referee)
The Smc5/6 complex is an eukaryotic protein complex that, together with Smc1/3 cohesin and Smc2/4 condensin, is involved in ensuring genome stability. It contributes to this by participating in the organization and maintenance of chromosomal structures as well as in the response to DNA damage. In addition, the Smc5/6 complex has been shown to play an important role in viral infection. This thesis focuses on the mechanisms of interaction of the Smc5/6 complex with viral DNA genomes, DNA intermediate genomes and viral proteins. In the case of HBV of the Hepadnaviridae family, Smc5/6 acts as a restriction factor. The same is true for HSV-1 of the Herpesviridae family, viruses of the Papillomaviridae family and HIV-1 of the Retroviridae family. An interaction of the Smc5/6 complex with the JC virus of the Polyomaviridae family has also been discovered. Nevertheless, the meaning of this interaction remains elusive. Some of the above-mentioned viruses can prevent this restriction. In detail, HBx protein of HBV mediates proteasomal degradation of the Smc5/6 complex or Vpr protein of HIV-1 induces degradation of the SLF2 protein, which is responsible for the Smc5/6 localization on HIV-1 DNA intermediate genomes. Keywords: Smc5/6 complex, DNA repair, ATPase, sumoylation, DNA viruses, viruses with a DNA...
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Role of molecular chaperones Hsp70 and Hsp90 in the replication cycle of DNA viruses
Žáčková, Sandra ; Horníková, Lenka (advisor) ; Poláková, Ingrid (referee)
Molecular chaperones are proteins which enable other proteins to assemble into native conformation and are essential for viability of the cells. Chaperones of the Hsp70 family bind to newly synthetized and denaturated proteins, prevent their aggregation and facilitate their assembly. They participate in assembly and disassembly of oligomers and also in the transport across the membranes. Chaperones of the Hsp90 family do not participate in the assembly of nascent or denaturated proteins. They bind proteins which are nearly in native conformation and enable them to assemble into conformation suitable for ligand binding or interacting with other proteins. These attributes predestinate chaperones to participate in the replication cycle of DNA viruses. A huge amount of proteins is translated during viral infection. These proteins require the chaperones to facilitate their assembly and are also required for assembly into oligomers and macromolecular structures. In addition to capsid assembly the chaperones also participate in transport of genetic information to the sites of replication, disassembly of incoming viral particles or replication of viral DNA. Therefore, the development of specific chaperone inhibitors is a promising approach. They could be used against broad spectrum of viral infections...
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