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Pentraxin PTX3 - a new marker of immunopathological inflammation.
Kondělková, Kateřina ; Jílek, Petr (advisor) ; Andrýs, Ctirad (referee)
Pentraxin 3 (PTX3) is a newly identified acute phase reactant which shares some structural and some functional properties with CRP, classical short pentraxin. On the other hand, PTX3 displays unique biological properties of its own, including a possible role in the pathogenesis of cardiovascular diseases and in processes accompanying the natural evolution of surgical wounds. Unlike CRP, which is manufactured predominatly in the liver, PTX3 is produced especially at sites of tissue damage. PTX3 synthesis is induced by proinflammatory cytokines TNF-α or IL-1β and by microbial wall constituents such as the lipololysaccharide (LPS). Plasma protein PTX3 concentrations are elevated in a wide range of diseased states, such as in coronary artery disease, in pulmonary infection and acute lung injury, 3 in patients with chronic kidney disease, in pacients with depression, during normal pregnancy and preeclampsia, in pacients with psoriasis and during cardiac surgery with or without cardiopulmonary bypass. PTX3 was detected using detection set (Alexis Biochemicals, Switzerland) cat.no. ALX- 850-299-KI01 for sandwich ELISA application that provided capture monoclonal antibody to PTX3, detection polyclonal antibody to PTX3 and recombinant PTX3 (standard). Goeckerman's therapy of psoriasis is highly efficient in...
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Immunological laboratory diagnostics of non-specific gut inflammation.
Vohralíková, Jana ; Jílek, Petr (advisor) ; Andrýs, Ctirad (referee)
BACKGROUND: Inflammatory bowel disease (IBD), with Crohn‛s Disease (CD) and ulceratice colitis (UC) as the two main disorders, is a heterogenous group of disease of unknown etiology. IBD serological markers may be important in the diagnosis of IBD, for differentiating CD from UC. The major serological marker in commonal use are ASCA and ANCA. Recently new antiglycan antibodies have been identified: anti laminaribioside carbohydrate antibodies - ALCA and anti chitobioside carbohydrate antibodies ACCA. The antibodies are significantly associated with CD. AIMS: To evaluate the significance of the recently discovered novel antibodies: ALCA and ACCA METHODS: Assesment of pANCA, ASCA, ALCA and ACCA was performed using the standardized indirect immunoflourescence technique and ELISA. Serum samples were obtained from 89 pacients with CN and from 33 pacients with UC. Results were considered positive if: ASCA> 15 U/ml, ALCA> 60 EU and ACCA> 90 EU. RESULTS: ASCA was positive for 78.8% (70/89) of patiens with CD and 12,1% (4/33) of pacients with UC. pANCA was positive for 42.4%(14/33) of patiens with UC and 7,8%(7/89) of patiens with CN. The ASCA IgA test yielded 71.3% sensitivity and 93.9% specificity for CN. The ASCA IgG test yielded 57,3% sensitivity and 93.9% specificity for CN. The ANCA test yielded 41.9%...
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Antimicrobial peptides in infections with grammnegative bacteria.
Tocháčková, Petra ; Jílek, Petr (advisor) ; Andrýs, Ctirad (referee)
Bc. Petra Tocháčková Antimicrobial peptides in infections with grammnegative bacteria Diploma thesis - abstract Charles University in Prague, Faculty of Pharmacy in Hradec Králové Pharmacy Background: Aim of this thesis was a development of methods for detection of gene activation of antimicrobial peptides (beta defensins 1 and 2 - pBD1 and pBD2) in ileum during infection of gnotobiotic piglets with gramnegative bacteria. Salmonella enterica serovar Typhimurium strain LT2 and its R mutants (rfaG- and rfaL-) were selected as representants of gramnegative bacteria of different virulence. Methods: 1) Suggestion of systems for detection of pBD1 and pBD2 (software), 2) Purification of total RNA (commercial columns), quantification (spectrophotometry), and estimation of its purity (ratio A280/A260), 3) Reverse transcription (mixture of random n-mers and oligo d(T)m), 4) Quantification of cDNA (real time polymerase chain reaction - LNA probes), and 5) Normalization of gene transcription for pBD1 and pBD2 against beta actin, cyclophilin A or to both together. Results: We used a method reducing risk of contamination by genomic DNA and obtaining of false positive results for purification of total RNA. We suggested detection systems for estimation of pBD1 and pBD1 with efficiency approx. 100% (comparable with...
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The effect of Goeckerman therapy on selected immune markers in patients with psoriasis
Kondělková, Kateřina ; Andrýs, Ctirad (advisor) ; Posová, Helena (referee)
1 Summary The effect of Goeckerman therapy on selected immune markers in patients with psoriasis Psoriasis is a systemic inflammatory disorder with skin manifestation. Classical approach to treat psoriasis is Goeckerman therapy (GT) which is based on daily application of pharmaceutical coal tar on affected skin with subsequent exposition to UV light. The aim of this study was to observe selected immunological markers and cell subpopulations in patients with psoriasis and to elucidate the effect of GT on these parameters. We evaluated T regulatory cells (Treg), membrane CD163 expression on monocytes, serum levels of CD163 (sCD163), membrane TLR2 and TLR4 expression on monocytes and serum levels of TLR2 (sTLR2). The immunofluorescent analysis was performed on erythrocyte lysed peripheral blood using a flow cytometer FC500 Cytomics (Beckman Coulter) and FACSCalibur (Becton Dickinson). The level of soluble markers was detected by sandwich enzyme-liked immunosorbent assay (ELISA). There was no significant difference in the relative number of Treg cells in the peripheral blood of healthy blood donors and patients with psoriasis before initiation of GT. The relative number of Treg is significantly increased in psoriatic patients after GT compared to the pre-treatment level. In comparison with levels in controls...
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