|
|
Determination of the concentration of alpha-1-antitrypsin in the stool by immunoassay method
Plačková, Marie ; Kocna, Petr (advisor) ; Průša, Richard (referee)
Determination of the concentration of α1 - antitrypsin in the stool is a diagnostic indicator of inflammatory diseases of the small and the large intestine, especially malabsorption syndrome. α1 - antitrypsin belongs to the family of plasma proteins with antiproteinase effect. α1 - antitrypsin is synthesized in liver, in small amount in macrophage and is a protease inhibitor of serine proteases sercected from neutrophils. α1 - antitrypsin is acute phase protein. Higher α1 - antitrypsin values are in early phase of inflammation associated with raised CRP and other pozitive acute phase proteins. Fecal α1 - antitrypsin clearance is a sensitive and specific marker of protein loss. For α1 - antitrypsin determination in stool samples ELISA method can be used. ELISA is noncompetetive immunoassay used to detect presence of antibody or an antigen in a sample. The aim of this work was to compare two ELISA sets (Immundiagnostik and Ridascreen) used for determination α1 - antitrypsin in the stool. Then examine stability of α1 - antitrypsin in the stool and in extract prepared from stool in various storing conditions temperature and time. After this establish this method as routine in laboratory. 20 patient stool samples were examined to compare ELISA sets. Samples were suggested to be α1 - antitrypsin...
|
|
|
Individual ....
Tomeček, David ; Kocna, Petr (advisor) ; Průša, Richard (referee)
13 C-MTG breath tests are used for exocrine pancreas diagnostics. It is a quite expansive test and its usage has never been tested in clinical practice in long time horizon. This study is engaged in pre-analytical phase and contains a group of 4 people who were tested for 6 months and this is the first study ever that has been proceeded in a long time horizon. The half-year profile of these 4 people showed that some of those results are spread of tens percents against its average. The test of CO2 stability showed that the value of CO2 is independent on the temperature which the breath bag is stored in. Its concentration was same for the time of 7 days. Now we can say that the breath bags are insensitive in case of transporting and storage. The question is, why the value of delta 13 C/12 C changed. The results of this study point on fact that values of cumulative dosage recovery of these 4 people did change.
|
|
|
Examining the significance of peptides regulating the intake of food and the nutritional state of children and adolescents
Bronský, Jiří ; Průša, Richard (advisor) ; Pelouch, Václav (referee) ; Hyánek, Josef (referee)
Introduction: Regulation of food intake and nutritional status is mediated by complex interactions of regulatory peptides of the central nervous system, gastrointestinal tract and adipose tissue. These systems are connected by feedback loops which inform the centre about amount of ingested food and energy reserves in the organism. Dysfunction of any of these regulatory areas may lead to changes in nutritional status of the organism. Methods: We used radioimmunoassay to measure plasma levels of orexin A, total ghrelin and serum levels of leptin and enzyme immunoassay to measure serum levels of adiponectin in healthy subjects and in children with obesity, anorexia nervosa, Crohn's disease and celiac disease and we evaluated the influence of nutritional therapy on these levels. Moreover, we evaluated relationship of these regulatory peptides to other biochemical and anthropometrical factors of nutritional status. We also measured plasma levels of total and unreduced amylin by enzyme immunoassay with immunofluorescence detection in adult patients with osteoporosis, type II diabetes mellitus and in the control group. Results: During reduction of body weight in obese children and adolescents, there were statistically significant changes of plasma orexin A levels and total ghrelin levels, but we haven't seen any...
|
|
|
Biochemical diagnostics of expansive brain contusions
Vajtr, David ; Průša, Richard (advisor) ; Bouška, Ivan (referee) ; Zima, Tomáš (referee)
TBI is one of the major causes of morbidity and mortality. Outcome for brain-injured patients is determined by the type, severity, location of the injury, ischemia and inflammatory response following TBI. Aim of the study: We focused on the relevance of a risk factor, kinetics of all markers in the blood reflecting different types of pathophysiological changes of the BBB in focal and diffuse injuries. We analyzed the BBB impairment in focal and diffuse brain injuries by immunohistochemistry and electon microscopy. Methods: The patients (n=38) were divided into group of diffuse axonal (DAI, n=10) and focal (n=28) brain injuries. Blood samples were collected during 10 days after admission to the hospital. Serum protein (S-100B, NSE) concentrations were measured on immunoassay analyzer Roche Cobas e411, GFAP by Biotrak activity assay systém, NF-H by ELISA immunoassay, and inflammatory markers on immunoassay analyzer. The BBB was studied by immunohistochemistry and electon microscopy. Results: With regard to the kinetics of NSE and S-100B we found the decrease of value of NSE and S-100B protein in survived patients with focal brain injury, on the other hand the increase of NSE (32,72 μg/l up to 86,95 μg/l) and S-100B protein (3,41 μg/l up to 5,80 μg/l) 2 up to 3 day was observed in 5 patrients who died. Within...
|
|
|
Identification and determination of metalloproteins and metalloproteinases in experimental models and in groups of patients
Kukačka, Jiří ; Průša, Richard (advisor) ; Zima, Tomáš (referee) ; Stoklasová, Alena (referee)
Introduction: Matrix metalloproteinases (MMPs) are zinc-containing metalloproteins which take part in many processes associated with extracellular matrix (ECM) remodeling. These enzymes participate in most processes degrading connective tissue during ontogenesis. Changes in MMP expression and activity can be observed in most inflammato ry, degenerative, and malign processes. Methalothionein also belongs to the group of zinc-dependent metalloproteins and it is involved in metal trafficing, detoxification or protection of cells against reactive oxygen species. Aim: The aim of th is study was to analyze MMP and metallothionein in various experimental models of caridac tissue remodeling (under hypoxia and methamphetamine administration) and in groups of patiens with dislipoproteinemia and traumatic brain injury. Methods: Laboratory rats were exposed to 1) hypoxia for 3 weeks and treated with MMP inhibitor 2) hypoxia and hypoxic hypercapnia for 4 days. 3) ln another experimental set methamphetamine was aplicated to rats for 9 weeks. 4) A group of patiens with dyslipoproteinemia was investigated before and after 1 month therapy by diet or hypolipidemics. 5) Blood samples were collected from patiens with traumatic brain injury during hospitalization. MMPs were analysed by zymography, immunochemical method, and...
|
|
|
Significance of determination SAA of comparison with CRP and procalcitonin
Daňková, Michaela ; Zadina, Jiří (advisor) ; Průša, Richard (referee)
The purpose of the bachelor thesis is to evaluate serum amyloid (SAA) as a diagnostic marker in selected patient groups and to compare the results of SAA with those of C-reactive protein (CRP) and procalcitonin (PCT). SAA, CRP and PCT are acute-phase proteins, whose blood concentration significantly increases within a few hours after an inflammatory stimulus. The synthesis, caused by proinflammatory cytokines, takes place in the liver. CRP is the most and longest used acute-phase reactant in routine practice. As indicators of inflammation, SAA and especially procalcitonin have been used more lately. Although SAA is an acute-phase reactant as sensitive as CRP, it is not employed so often in the diagnosis since a method suitable for routine diagnosis was not developed until recently. Briefly describing the acute-phase proteins, the theoretical part elaborates on the characterization and comparison of SAA, CRP and PCT. It further outlines methods for the determination of individual analytes. The experimental part focuses on defining selected patient groups and the immunonephelometric method for measuring SAA by the Immage 800 biochemical analyzer. The data obtained in three selected patient groups have been statistically evaluated. The aim of the thesis has been to evaluate the correlation of inflammatory...
|
|
|
Immunoflulorescent detection of endomysial antibodies (EmA) in the coeliac diagnosis
Martikán, Petr ; Kocna, Petr (advisor) ; Průša, Richard (referee)
The aim of this thesis was comparing of a several immunofluorescence methods and substrates for detection endomysial antibodies (EmA). The common method (Anti-Endomysium antibodies AEA) is used by UKLBD in VFN and by the some samples gives ambigious results owing to another autoantibodies (e.g. ANA, AMA, ASMA). The common method was compared with another sets (IFA-EmA-IgA - AEMAk 03a-10, ImmuGlo anti Endomysial Antipody EMA IFA Kit). Another aim of the work was how to work with this method and be able to evaluate samples by the fluorescence microscope. (...) This thesis brought me a lot of valuable experiences in laboratory and also teached me how to evaluate endomysium antibodies by indirect immunofluorescence method.
|
|
|
Monitoring of biochemici markers of septic states
Brodská, Helena ; Kazda, Antonín (advisor) ; Zazula, Roman (referee) ; Průša, Richard (referee)
The topic of systemic inflammatory response is lately often discussed across the whole field of medicine. Early diagnosis and the following adequate therapy minimize the risks of the development of systemic inflammatory response syndrome (SIRS). Therefore there is a general urge to find an appropriate marker for diagnosing and monitoring of the development of SIRS. Each year new markers appear, markers that fill up the mosaic of a pathophysiological picture, but we still do not have a marker, that would fulfill the demands of specificity, sensitivity and availability. In the recent ten years, one of the most promising markers seems to be the PCT. (...) In two studies with patients in end states renal diseases (ESRD) the non infectious elevations appear as marker of severity of clinical state. In the last study we confirm the relationship between PCT and severity of clinical state and the antioxidative defence, respectively the plasmatic selenium level. In the above mentioned studies I would like to present the wide range of this marker, which in the meantime is one of the most used markers of the SIRS. I would like to stress the importance of a correct indication and of an accurate interpretation in accordance with all the other parameters, but most of all with the clinical state. PCT, being almost ten...
|
|
|
Investigation of thyroid disorder in pregnancy and reference intervals in evaluation of maternal thyroid function
Springer, Drahomíra ; Zima, Tomáš (advisor) ; Průša, Richard (referee) ; Zamrazil, Václav (referee)
The importance of maternal thyroxine for the development of the fetus brain early in pregnancy has received increasing acceptance. It has more recently become evident that maternal hypothyroxinemia results in the birth of children with decreased mental and psychomotor development. In our group of 7,530 women in 9-11 week of pregnancy were determined TSH, anti TPOAb and FT4. For evaluation of results was necessary to set reference intervals for pregnant women. The TSH reference interval was determined to be 0.06 - 3.67 mU/l and for FT4 9.8 - 23.43 pmol/l was used. The limit for anti-TPO positivity was determined to be 143 kU/l. A raised concentration of TSH was found in 5.14% of women; and a suppression of TSH was found in 2.90% of women and 11.5% of pregnant women were found positive. Serum concentrations of FT4 were lower in TPOAb positive as compared to TPOAb negative women and differences of FT4 in euthyroid women with suppressed, normal and elevated TSH were found. (...) In Czech Republic, case finding screening is able to disclose only about 20% of asymptomatic mild or deep hypothyroidism or women with positive anti TPO in pregnancy. Foreign study report 70% of these high-risk pregnant women, consequently is evident need of general screening of pregnancy thyroid failure in Czech Republic. Moreover the...
|
|
|
Oxidative and carbonyl stress in kidney diseases
Kratochvílová, Markéta ; Tesař, Vladimír (advisor) ; Průša, Richard (referee) ; Zadražil, Josef (referee)
Aims: 1. Determination of AGEs (Advanced Glycation End products) in patients with various types of nephropathy. 2. Association AGEs with nutritional parameters and anemia. 3. Influence of renal parameters on sRAGE (soluble form of Receptor for Advanced Glycation End products) levels. 4. Technics and proceeding methods of the podocytes cultivation. 5. Determination of urine podocytes. Methods: We determined fluorescent AGEs by spectrofluorometry, sRAGE by Enzyme-Linked ImmunoSorbent Assay (ELISA). Podocytes were passaged and identified immunocytochemically. Podocytes in urine were specified by flow cytometry method. Results: 1. We did not find significant differences in AGEs serum levels among various types of nephropathy, even though the pathogenesis differs. 2. The albumin and prealbumin levels positively and haemoglobin levels negatively correlate with AGEs in patients with CKD grade 1-5, without necessity of dialysis. 3. Serum sRAGE levels are increased in patients with decreased renal function independently on the course of renal disease. 4. We implemented the methods and technics of podocyte cultivation. 5. Urine podocytes observation and confirmation that podocyturia relates to disease activity. Conclusion: We confirmed that AGEs serum levels depend more on renal function than the type of...
|
guest ::
RSS feed.