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The study of bacterial microflora at patients with spontaneous early delivery.
Pasdiorová, Markéta ; Hronek, Miloslav (advisor) ; Vrbacký, Filip (referee)
1. ABSTRACT Bc. Markéta Pasdiorová The study of bacterial microflora at patients with spontaneous early delivery Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Healthcare Bioanalytics - Specialist in Laboratory Methods Backround: Premature delivery and premature rupture of membranes have been associated with maternal genital tract infections or with altered vaginal microflora during pregnancy. This work discuses knowledge about vaginal microbiome, premature delivery and analysis options of vaginal microbiome using next-generation sequencing. The aim of the work was to evaluate the composition of the bacterial microflora at patients with spontaneous preterm delivery and compared with each other of vaginal and cervical samples. Methods: The study group consisted of 7 patients with diagnosed spontaneous preterm delivery. Samples were obtained on the basis of cooperation with the Department of Obstetrics and Gynecology of University Hospital Hradec Králové. The evaluation of the vaginal microbiome was performed by pyrosequencing method using Genome Sequencer FLX+. Results: We have observed all typical patterns of the vaginal microflora in the samples tested. The most frequent species were Lactobacillus crispatus/casei and L. iners, which belong to the four most dominant...
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BRAF mutations in metastatic malignant melanoma.
Hrabcová, Veronika ; Hronek, Miloslav (advisor) ; Vrbacký, Filip (referee)
Bc.Veronika Hrabcová BRAF mutations in metastatic malignant melanoma. Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Healthcare bioanalytics - Specialist in Laboratory Methods Backround: Melanoma is malignant disease with increasing incidency. Treatment of advanced stage of melanoma is still limited. With a progress of knowledge in genetics and tumorigenesis, the incidence of mutated BRAF protein was observed at 50 % of melanomas. In 80-90 % mutated melanomas contain BRAF V600E mutation. The aim of study was to establish a suitable molecular biological method for the diagnosis of mutations in codon V600 BRAF. Methods: Cobas 4800 BRAF V600 mutation test and BRAF StripAssay test were used to analyze DNA. Cobas 4800 BRAF V600 mutation test is based on PCR using TaqMan probes designed for the wild-type and mutant BRAF V600E sequence. BRAF StripAssay test is based on PCR amplification with biotinylated primers and subsequent hybridization of the stripped with allele-specific oligonucleotide probes. Examined DNA samples were derived from 35 patients with advanced malignant melanoma or from archive of laboratory. Results: BRAF V600 mutation was detected in approximately half of the tumors, consistent with the results of other studies. In comparison methods Cobas test...
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Comparison of differential leukocyte count set by the analyzer Beckman Coulter LH 750 and by microscope in the group of pediatric hemato- oncology patients
Šturmová, Michaela ; Fátorová, Ilona (advisor) ; Vrbacký, Filip (referee)
In my thesis I focused on comparing a differential leukocyte count (DIF) determined by the analyzer and by microscope in the group of pediatric oncological and hemato-oncological patients. Thesis is divided into theoretical and practical parts. The theoretical part includes an explanation of the process of hematopoiesis, the development of blood cells and the characteristic of time series, a decription of hemato-oncological and oncological diseases which occur most frequently in the monitored group of pediatric patients, and their treatment. In a practical part is described a determination of the DIF by the analyzer Beckman Coulter LH 750 and by microscope Meopta. I measured a blood count with the differencial leukocyte count, I made a blood smear and I evaluated microscopically it, in the selected group of pediatric patients. The results were reported in the tables and statistically processed using paired t-test in GraphPad Prism and were created graphs. An aim of this thesis was to explore and evaluate further the differences between the two methods. In 19,32% of patients in the test group was not evaluated the DIF by the analyzer . They were patients with the fresh catch of acute leukemia, were they applied growth factors or were they the patients after chemotherapy. If in such cases analyzer...
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Detection CD56 antigenic expression of the patient with morfology report LGL in the peripheral blood.
Procházková, Irena ; Fátorová, Ilona (advisor) ; Vrbacký, Filip (referee)
Author: Bc. Irena Procházková Job title: Detekce exprese antigenu CD56 u pacientů s morfologickým nálezem LGL v periferní krvi Diploma work Charles Univerzity in Prague, Faculty of Pharmacy in Hradec Králové, Biological and medical department Field of study: Expert of specialized methods Abstract The goal of this diploma work was gathering evidence to support the theory, that LGL cells, large granular lymphocytes, are identical to the population of NK cells, that are detectable by antigen CD56. Numbers of the CD56+ cells were gained by flow cytometry measuring the expression of antigen CD56 labeled with antibody against CD56. The correlation was reviewed with the numbers of LGL cells, which were found in the periferal blood of the patient by microscopic evaluation with common panoptical staining of blood coat. There were used the absolute counts of detected parameters for this comparison. The leukocytes from the measured blood count of the patients on the analyzer Advia 2120i were used for calculation the absolute counts of LGL cells. In the group of patients with immunodeficiencies and various forms of allergy where established an increased number of CD56+/CD3- cells (NK cells), we performed a correlation with the number of LGL cells defined microscopically. We found a strong correlation betwwen the two...
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Laboratory diagnostics of multiple myeloma.
Hubálková, Blanka ; Kovařík, Miroslav (advisor) ; Vrbacký, Filip (referee)
Multiple myeloma is one of the main representatives of monoclonal gammapathy diseases. It is caused by malignant transformation of B-lymphocytes, its clonal proliferation and accumulation of terminal stages of plasmocyte maturation. Clinical diagnosis is based on presence of this monoclonal immunoglobulin in serum or urine, myeloma cells infiltration into bone marrow and finding osteolytic lesions in bones. The electrophoresis and immunofixation of monoclonal immunoglobulins, free light chains assay, cytology of bone marrow aspirate are the most important laboratory tests helping in the statement of this immunoglbuline. The proof of the osteolytic damage is made using imaging technologies, such as RTG. Treatment of multiple myeloma had significantly improved during last years. The main options of therapy include high-dose chemotherapy and autological transplantations, together with immune modulating drugs, such as thalidomide, lenalidomide and bortezomib. Experimental part was focused on the comparison of sensitivity of paraprotein assessment by electrophoresis, immunofixation and free light chains assay. We can assume on the base of the results of the work that the free light chain assay is less sensitive method compared to k/I index. In addition - the higher sensitivity of IFO in comparison with...
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Morphology and cytochemical diagnosis of hairy cells leukemia
Bergerová, Lenka ; Fátorová, Ilona (advisor) ; Vrbacký, Filip (referee)
5 Title: Morphology and cytochemical diagnosis of hairy cells leukemia (Bachelor thesis) Author: Lenka Bergerová Department: Charles University in Prague - Faculty of Pharmacy in Hradec Králové, branch of study Medical Laboratory Technician Supervisor: MUDr. Dana Mikulenková Abstract Hairy cell leukemia is the chronic lymphoproliferative disorder of clonal character arising from mature B cells. In the peripheral blood smear and in the bone marrow smear, we find the typical leukemic cells which are characteristic by their size, morphology of cell nucleus and cytoplasm. They are bigger than healthy lymphocytes, and they have most frequently the kidney shaped nucleus with fine net-like chromatin and the cytoplasm projecting into fine salients all over the cell perimeter. In the blood count we usually find a mild pancytopenia. To demonstrate the presence of the pathological cells in the peripheral blood smear and the bone marrow smear, we use the panoptic staining method according to Pappenheim. Among other examinations contributing to the diagnostics of this disease belongcytochemical examination of acid phosphatase and acid phosphatase resistent for tartarate, immunophenotypic findings and trepan bone marrow biopsy. In addition to the typical form of hairy cell leukemia we distinguish another variant form of...
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Laboratory diagnostics of anemias
Halamková, Renata ; Fátorová, Ilona (advisor) ; Vrbacký, Filip (referee)
1 ABSTRACT This bachelor theses deals with anaemia, one of the most common hematological diseases. The introductory part of this work defines the possible causes of this disease and its clinical picture. The following part includes the logical division of different types of anaemia accompanied with examples of real cases. Main focus of the theses was put on sideropenic and megaloblastic anaemia, as they occur considerably in the population.
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Search for a suitable housekeeping gene for relative quantification in patients with the chronic lymphocytic leukemia
Ryšková, Barbora ; Vrbacký, Filip (advisor) ; Sadílek, Petr (referee)
5 ABSTRACT Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Barbora Ryšková Supervisor: Mgr. Filip Vrbacký Title of diploma thesis: Search for a suitable houskeeping gene for relative guantification in patients with the chronic lymfocytic leukemia Chronic lymphocytic leukemia (CLL) is the most common form of leukemia found in adults and that is why it is studied very intensively. The cause of CLL is still unknown and it is not known how to cure this lyphoproliferative disorder without high risk of complications that may occur with a bone marrow transplantation. Altered gene expression of angiogenic factors in CLL is still insufficiently known too, so we decided to study this topic in our laboratory. Our task was to establish method for quantification of proper housekeeping gene that can be used for normalization of results of quantification of expression of some angiogenic factors by real-time PCR. We decided to use Abl1 gene but that gene is alternatively spliced. Exon 1 containing variant is housekeeping only and we had to take it into account when choosing proper method. I managed to transform Escherichia coli cells and amplify plasmid containing Abl1 sequence. Plasmid was isolated, quantificated by spectrophotometer and used to...
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