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Molekulární diagnostika fungálních patogenů z klinických vzorků
CIMICKÁ, Jana
This study is focused on optimization and use of molecular methods in mycological diagnostics. The patient's life often depends on early diagnosis, which current culture methods cannot provide. The solution is based on pan-fungal primers of ITS region and next generation sequencing (NGS). These methods were used for fungal detection from FFPE tissue samples from patients with ulcerative colitis (UC), Crohn disease (CD) and 10 controls without chronic inflammatory changes in the intestine. Results of this study have three levels. Firstly, the process of optimization highlights the importance of sterile DNA isolation in a laminar box which can prevent contamination by Malassezia spp. Secondly, raw data showed balanced rate of Ascomycota and Basidiomycota in CD samples similar to negative controls, while UC samples indicated higher representation of Ascomycota than other groups. Thirdly, detailed data showed fungal genera Malassezia, Cladosporium and Toninia in all groups, while genus Candida was found only in UC samples and genera Engyodontium and Ramularia were found in CD samples. In conclusion, mycobiome plays a role in inflammatory bowel diseases; however, its compartments are still questionable. The optimization of pan-fungal nested PCR and NGS helped with introduction of mycological diagnostics from FFPE samples to Bioptická laboratoř s.r.o. and is used for differential diagnostics.
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Inflammatory Bowel Diseases in pregnancy
Kovářová, Tereza ; Krátká, Zuzana (advisor) ; Hadlová, Petra (referee)
(EN) Inflammatory bowel disease is a chronic autoimmune disease of the intestinal tract including Crohn's disease, ulcerative colitis, and other unspecified colitis. Depending on the type of disease, inflammation affects the small or large intestine or rectum. It is a multifactorial disease in which the intestinal microflora, the immune system status, and genetic predisposition are crucial. IBD is most often diagnosed in late adolescence or early adulthood. Due to the availability of quality treatment, the health status of IBD patients also improves, allowing them to consider starting a family after the disease has calmed down. Patients have fewer children than the general population, but there was no significantly higher risk of infertility. A prerequisite for successful pregnancy and childbirth is a stable condition of the disease and the use of treatment during pregnancy to prevent relapse. A specific problem of patients is voluntary childlessness because of worries about the worsening of IBD during or after pregnancy, about negative effects of treatment on a foetus, but also an occurrence of disease in offspring This work aims to summarize the issue of pregnancy in patients with IBD and to provide information on the current situation in patients with IBD in the Czech Republic. Keywords:...
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Changes in distribution of B lymphocyte subpopulations in patients with Crohn disease before and after biological therapy
Suchá, Renata ; Růžičková, Šárka (advisor) ; Krulová, Magdaléna (referee)
B-lymphocytes are lymphoid cells, which are a part of the adaptive/innate immune system and generate antibodies. Recently, many studies have supported hypothesis that different rather minor B-lymphocyte subpopulations may play a direct and indirect role in immunopathogenesis in human pathologies such as Crohn's disease (CD). The aim of current study was therefore to investigate distribution of frequencies of B lymphocyte subpopulations (from transient to mature effector B cell stages) in peripheral blood of healthy subjects (CO), patients with Crohn's disease (CD) and ulcerative colitis (UC). Thus, using 11-colour flow cytometry we have analysed 30 blood samples of individuals, including 14 healthy controls, 11 patients with Crohn's disease and 5 with UC. In 6 patients with CD we have had an opportunity to analyze blood samples collected 2 hours after an administration of anti-TNF therapy. Higher frequencies of memory B-lymphocytes (CD19+ CD27+ , CD19+ CD20+ CD27+ and CD19+ CD20+ CD27+ IgM+) were found in patients with CD as compared to COs. (20.06±13.58%; 17.61±13.48%; 88.60±20.56% vs. 11.75±26.47%; 11.25±26.50%; and 66.82±22.60%), in case of CD19+CD20-CD27-IgM+ B-lymphocytes the difference was statistically significant (57.15±17.21% in CD vs. 19.59±31.79% in CO; p=0.0341), which is in accordance...
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Quality of life in patients with inflammatory bowel diseases
Píchová, Andrea ; Mellanová, Alena (advisor) ; Marková, Eva (referee)
Quality of life in patients with inflammatory bowel disease (IBD) depends on activity, therapy and complications of this chronic disease. Further more it also depends on psychosocial and economic aspects. Nurse plays an important role in education and psychological care for these patients. Aim of this work, based on questionnaire and its statistical processing is to evaluate quality of life of IBD patients (according to age, education and social background) and monitor its dynamics from the time of diagnosis and during treatment of the disease. 1. -6-
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