|
. Changes in gene expression of B cell line Ramos RA-1 after infection with Francisella tularensis.
Matyášová, Simona ; Konečná, Klára (advisor) ; Kubelková, Klára (referee)
Francisella tularensis is a facultative intracellular bacterium causing disease tularemia of different species of mammals, including humans. It is able to infect phagocytotic as well as non-phagocytotic cells. We focused on the study of B cells, that role in the infection of F. tularensis have been, until recently, regarded as not significant. Over the last 10 years several studies showed important role of B cells in innate immunity to F.tularensis infection. The aim of this thesis was to idenify changes in the expression of genes that encode molecules of cytokines produced by B cells, after F. tularensis infection, due to stimulation with bacterial recombinant protein IglC and lipopolysaccharide. These changes were examinated on B cell line Ramos RA-1. Using reverse transcriptase polymerase chain reaction, we detected these cytokines: IFN-γ, IL-10 a TGF-β. Key words: Francisella tularensis, B cells, cytokines, reverse transcriptase polymerase chain reaction
|
|
Chronic Obstructive Pulmonary Disease ant its Treatment
Vrátná, Sandra ; Vopršalová, Marie (advisor) ; Žáčková, Pavla (referee)
1 ABSTRACT The aim of my thesis was a comprehensive view of COPD as a separate nosological entity. A substantial section focuses on current therapy, which are listed by category as concessional and stability of drugs including their mechanism of action. Education and smoking prevention play major role during the non- pharmacological therapy , as well as the rehabilitation, which is not yet very common, but it may contribute significantly to patientś health improving. The work is also illustrative of the disease compared with bronchial asthma because of the significant similarities of these diseases, their possible replacements and overlays number of syndromes that may complicate the diagnosis of any disease. A separate chapter is devoted to innovations in treatment, which lead to new possibilities in the combination of drugs for immunotherapy, but also necessary for long-acting drugs that promote patient compliance and is seeking to increase their comfort during therapy. Given that most drugs are administered by inhalation, indicated in my work with the issue of inhalation systems, their separation and the available tools offered by the Czech market.
|
|
Frequency of multiple head and neck tumors on department of otorhinolaryngology of FNKV hospital from 1984 to 2009
Cimalová, Lenka ; Čoček, Aleš (advisor)
The incidence of malignant tumors of head and neck is rising worldwide and the incidence of subsequent neoplasms in this area is increasing as well. Diagnosis can be difficult. Most patients underestimate symptoms and a large proportion of disease is diagnosed at a late stage. The common feature of head and neck tumors is their histological composition (90-95% are squamous cell carcinomas with varying degrees of differentiation) and a tendency to locally spread with the early emergence of locoregional metastases. The group most at risk is men over 45 years old, especially cigarette smokers and drinkers - especially spirits. Other risk factors include infection of HPV and HSV viruses, occupational pollution, poor nutrition, poor oral hygiene and inadequate dental prosthesis and so on. The role of prevention seems to be essential in case of head and neck cancer. According to the American Cancer Society 85-90% of head and neck tumors is caused by tobacco products. This is one of the reasons why the fight against smoking has become a major global prevention effort. The crucial role in the treatment of these tumors has a surgical treatment and radiotherapy, but chemotherapy has just a limited effect. The future way of therapy is likely to be a biological treatment. The multiple tumors mean subsequent primary...
|
|
delta subunit of bacterial RNA pol and its role in regulation of gene expression in B. subtilis
Dvořáček, Lukáš ; Krásný, Libor (advisor) ; Vopálenský, Václav (referee)
Delta subunit of bacterial RNA pol and its role in regulation of gene expression in B. subtilis. In this work I focus on regulation of eubacterial gene expression. First, I describe recent knowledge about a key stage of gene expression - transcription, focusing on regulation of trancription iniciation via small effector molecules (guanosine tetraphosphate, initiating nucleoside triphosphate) that are important for the regulation of ribosomal RNA. Second, in the experimental part of my work, I focus on the role of the _ protein, a subunit of RNA polymarase in gram positive bacteria, in transcription iniciation and its effects on regulation of RNA polymerase by the concentration of initiating nucleoside triphosphates.
|
|
Significance of prolactin as peripheral cytokine in dysbalance of immune system
Janatová, Kateřina ; Černá, Marie (advisor) ; Holáň, Vladimír (referee)
Background: Interactions between the neuroendocrine and immune system play an importatnt role in maintaining homeostasis. This communication is mediated by cytokines, neurotransmiters and hormones through endocrine, paracrine and autocrine signaling. Prolactin (PRL), hormone of anterior pituitary, is produced by a number of other tissues and cells of immune system. On periphery, PRL is cytokine. Sepsis is an inflamatory response of the organism to severe infection, Th1 immune response is activated and PRL could participate in it. Toll-like receptors (TLR) play a key role in a recognition of bacteial components and mediate a systemic response (with PRL secretion) during infection. It is supposed that activated immune system leads to increasing of PRL, TLR2 and TLR4 gene expression. We detected PRL, TLR2 a TLR4 mRNA levels in monocytes from patiens with system inflammation. We studied influence of single nucleotide polymorphism (SNP -1149 G/T) in PRL gene promotor, it supposed that G allele increases PRL expression. Materials and Methods: For the pilot study 30 patients diagnose with severe infectious event. Collectoin of patiens blood samples was performed consequently three times. Control group comprised 40 healthy individuals. One blood sample was taken from each healthy subject. For testing of...
|
|
Domain conformations of the motor subunit of EcoR124I involved in ATPase activity and dsDNA translocation
BIALEVICH, Vitali
Bacterial type I restriction-modification systems are composed of three different subunits: one HsdS subunit is required for identification of target sequence and anchoring the enzyme complex on DNA; two HsdM subunits in the methyl-transferase complex serve for host genome modification accomplishing a protective function against self-degradation; two HsdR (or motor) subunits house ATP-dependent translocation and consequent cleavage of double stranded DNA activities. The crystal structure of the 120 kDa HsdR subunit of the Type I restriction-modification system EcoR124I in complex with ATP was recently reported. HsdR is organized into four approximately globular structural domains in a nearly square-planar arrangement: the N-terminal endonuclease domain, the RecA-like helicase domains 1 and 2 and the C-terminal helical domain. The near-planar arrangement of globular domains creates prominent grooves between each domain pair. The two helicase-like domains form a canonical helicase cleft in which double-stranded B-form DNA can be accommodated without steric clash. The helical domain, probably involved in complex assembly, exhibits only a few specific interactions with helicase 2 domain. Molecular mechanism of dsDNA translocation, cleavage and ATP hydrolysis has not been yet structurally investigated. Here we propose a translocation cycle of the restriction-modification system EcoR124I based on analysis of available crystal structures of superfamily 2 helicases, strutural modeling and complementary biochemical characterization of mutations introduced in sites potentially inportant for translocation in the HsdR motor subunit. Also a role of the extended region of the helicase motif III in ATPase activity of EcoR124I was probed.
|
|
Molecular Markers with Impact on Kidney Graft Survival and Glomerulopathies Progression
Brabcová, Irena ; Viklický, Ondřej (advisor) ; Jirsa, Milan (referee) ; Rychlík, Ivan (referee)
The progression of chronic glomerulopathy and graft rejection is affected by a number of proinflammatory cytokines, whose role in the pathogenesis of damage is poorly understood. The aim of this dissertation was to identify reliable risk markers of renal dysfunction progression and thereby contribute to a more effective patient treatment. Human native kidney biopsies with histologically confirmed diagnosis of glomerulopathy or kidney graft biopsies were analysed. Intrarenal gene expressions were measured by RT-qPCR. Single nucleotide polymorphisms were detected by methods based on PCR-RFLP. Immunohistochemical staining was used to identify and quantify the mononuclear cell infiltration. Gene expression of TGF-β1, HGF, BMP7, MCP-1, RANTES and mononuclear cell infiltration were associated with poor renal function and proteinuria at the time of IgA nephropathy diagnosis. Progression of IgA nephropathy during the 2-year follow-up was shown to be dependent on the degree of chronic vasculopathy and TGF-1 expression in the kidney. Patients with graft dysfunction and enhanced intrarenal expression of TGF-1, MCP-1 had significantly shorter graft survival. Higher mRNA expression of IL-10, TGF- 1, IL-6, MCP-1, RANTES and TNF- was observed in patients with graft dysfunction presented at the time of biopsy....
|
|
Molecular Pathology of Rett Syndrome
Záhoráková, Daniela ; Martásek, Pavel (advisor) ; Kubala Havrdová, Eva (referee) ; Mazura, Ivan (referee)
Rett syndrome (RTT) is a severe X-linked neurodevelopmental disorder affecting almost exclusively girls. It belongs to autistic spectrum disorders and it is characterized especially by psychomotor regression, loss of acquired speech, microcephaly, repetitive stereotypic hand movements, and seizures. Most of RTT cases are caused by de novo mutations in the gene for the methyl-CpG-binding protein 2 (MECP2) and familial cases are extremely rare. The MECP2 gene product plays an important role in chromatin remodeling, regulation of gene expression and is also involved in RNA splicing. Some atypical RTT cases are caused by mutations in other genes, such as CDKL5, FOXG1 or NTNG1. In this paper we give an overview of RTT, its clinical aspects, molecular basis, diagnostic criteria, medical management and DNA diagnosis.
|
|
Polyhydroxyalkanoates and their role in bacterial biofilms
Rucká, Markéta ; Samek,, Ota (referee) ; Obruča, Stanislav (advisor)
This master thesis deals with polyhydroxyalkanoates (PHA) and their role in bacterial biofilms. In the theoretical part the polyhydroxyalkanoates, bacterial biofilm and the relationship between them were reviewed. The experimental part focused on differences in PHA production by planktonic and biofilm cells. In order to study selected topic, bacterial strains of Burkholderia cepacia and Burkholderia sacchari were cultivated using a CDC biofilm reactor. The attention was paid to quantity and especially to the form in which PHA occurs in planktonic and biofilm cells. Results of Raman spectroscopy have shown that PHA exists exclusively in native amorphous form in planktonic bacterial cells. On the other hand, in biofilm PHA occurs also in a partially crystalline form. In addition, the resistance of planktonic and biofilm cells against various stress factors and the effect of osmotic stress on PHA production was tested too. According to the results of the experiment, when the bacteria were exposed to different stress factors (high temperature, low temperature, presence of detergent and so forth) biofilm cells showed a higher stress resistance than planktonic cells. Apart from slowing cell growth and reproduction, increased osmotic pressure in the culture medium also caused decrease of PHA production. In addition, planktonic cells responded to external stimuli more sensitively than biofilm ones.
|
|
Problems of the functional flatfoot
Michalec, Radim ; Dudová, Agnieszka (advisor) ; Véle, František (referee)
Title: Problems of the functional flatfoot Objective: The aim of this diploma thesis is problems of the functional flatfoot. Main goal is to describe relation between change of structure, disorder of foot function and development of flatfoot. The thesis tries to describe the behavior of foot and arch in standing and walking. Another goal is to summarize the diagnostic possibilities of flatfoot. Methods: The diploma thesis has descriptive-analytical character. It's written in form of literature review. Results: This thesis demonstrates the problem of disunity nomenclature concerning the problem of flatfoot. Specifically, it concerns the interpretation of the words of functional flatfoot, movement components in foot and laboratory diagnostic methods. There is offered an explanation of the concept of functional flatfoot. Further work indicated that the acquired flatfoot is a question of a functional disorder that may progress to structural changes. The static and dynamic loadings were confirmed by the role of small muscles of the foot in the role of dynamic stabilization and influence of femoral rotation on the foot arch. There were summarized methods used for the diagnosis of flatfoot including dynamic plantography. Keywords: flat foot, foot arch, kinesiology, diagnostics
|