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The role of monocytes in pathogenesis of diabetes - immunogenetic study
Paukner, Karel ; Daňková, Pavlína (advisor) ; Černá, Marie (referee)
Type one diabetes is an autoimmune disease. It is caused by the destruction of β cells of Langerhans' pancreatic islets. Hyperglycemia is a major symptom of β cell destruction. Monocytes play a key role during T cell activation. T cell effect can be protective (Treg) or destructive. Monocyte destroys β cells as a macrophage and generates self-tolerance as a dendritic cell. The number of patients with T1D is increasing. In the presented work I aim to summarize current information about pathogenesis of T1D and I try to propose future way of research.
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Molecular aspects of calcium-phosphate metabolism in renal complications of diabetes mellitus.
Šimáková, Eva ; Černá, Marie (advisor) ; Zajíčková, Kateřina (referee)
Introduction: This thesis deals with the calcium-phosphate metabolism and its role in the development of chronic diabetic complications. It examines calcium sensitive receptor, which can be crucial in affecting calcium metabolism. Calcium-sensitive receptor may play a role in intracellular signaling and metabolic pathways that lead to cell proliferation and extracellular matrix early diabetic nephropathy. We investigated two polymorphisms of the gene for calcium-sensitive receptor (intron 4 and codon 990). Material and Methods: The study included 313 diabetic patients, 41 patients with type 1 diabetes, 106 patients with type 2 diabetes, 110 diabetic patients with type 2 diabetes who had diabetic nephropathy and 56 patients with renal failure non-diabetic (NDRD). It was also examined 72 non-diabetic patients with chronic renal failure (CKD) and 96 healthy blood donors (ZK). Classification of diagnoses is listed in the theory. DNA was isolated by QIAamp DNA Blood Mini Kit and salting method. The specific fragments of gene for the CaSR were amplified by PCR. For detection, restriction fragment length polymorphism and TaqMan probes were used. The expression levels of mRNA were determined by real-time PCR. Results: For the codon 990 polymorphism, we found statistical significance of the genotype frequencies (AA,...
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Změny exprese signálních proteinů a genů v souvislosti s diabetickou nefropatií
Demová, Hana ; Černá, Marie (advisor) ; Viklický, Ondřej (referee) ; Kalousová, Marta (referee)
Hypothesis: I have investigated specific molecular and cell mechanisms that might be involved in the ethiopatogenesis of diabetic nephropathy. Their involvement might also be demonstrated in response to treatment of diabetic nephropathy. Aims: I have studied changes of signal transduction proteins (CAV-1, VEGF, RhoA) in renal cortical cells in the rat model of renal hypertension and in the rat model of type 1 diabetes. I also have performed analyses of polymorphisms in the genes involved in cell signaling with respect to their function in human study. Methods : The renal cortical expression of molecular targets CAV-1, VEGF, and RhoA have been assessed in addition to measurements of renal functional parameters in L-NAME-treated rats with or without a concomitant administration of atorvastatin (ATO) and have been compared to untreated control animals.
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Assessment of expression markers of HLA class II genes.
Zajacová, Marta ; Černá, Marie (advisor) ; Černý, Jan (referee)
BACKGROUND: HLA molecules play a central role in the immune response. HLA class II are involved in the selection of the T-cell repertoire in the thymus, and in presentation of antigenic peptides to antigen reactive CD4-positive T cells. The HLA class II act as restriction determinants in the presentation of antigens to T lymphocytes and their expression on the cell surface is necessary for triggering the immune responses. Regulated transcription of HLA class II genes is a complex system involving cis-acting sequence elements and trans-acting protein factors. It has been reported that allelic polymorphism exists in the regulatory regions of HLA class II DQA1, DQB1 and DRB1 genes. Most of the polymorphisms appear to be conserved within a haplotype. The hierarchy of sequence homology which exists among the structural genes is not paralleled among their promoter sequences. It is of interest that the most striking discrepancy was found for the DR4 linked alleles, DQB1*0301 and DQB1*0302, which were described as frequent risk factors for a variety of autoimmune diseases. The structural genes of these two alleles are the most closely related, but their regulatory sequences are the most heterogeneous among DQB1 variants. These sequence differences correspond to functional variation: The promoter strength...
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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...
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Role of heat shock proteins in the pathogenesis of leukaemia
Kopřivová, Olga ; Hromadníková, Ilona (advisor) ; Černá, Marie (referee)
(Abstract) Some of heat shock proteins (Hsp), for example the inducible form Hsp70, are expressed on the surface of tumour cells. High Hsp expression is reflected in tumour cell features, such as ability to progression, to metastasize and resistance to apoptosis. The question is whether Hsp gene expression correlates with surface expression. The aim of this master thesis is to compare surface and gene expression of Hsp70 and observe the gene expression of some other Hsp proteins (Hsp27, Hsp60, Hsp90 and HspBP1) in leukaemia. The research was carried out on cell lines obtained from leukaemic blasts of patients with acute myeloid leukaemia: UoC-M1, HL-60, OCI/AML3, THP-1, HU-3 and TF-1 that had been cultivated in vitro. Hsp70 surface expression was detected using flow cytometry, and gene expression of each Hsp was studied using real-time RT-PCR. It was found out that high surface expression of Hsp70 did not correlate with gene expression in consequence of negative feedback applied in Hsp expression regulation. Hsp27 gene expression was increased compared to negative (healthy) control on all tumour cell lines, with the highest increase on the THP-1 line. Hsp60 gene expression was increased compared to negative (healthy) control on all tumour cell lines and there were not remarkable differences in...
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Genetic, humoral and cellular factors of development of autoimmune diseases
Růžičková, Šárka ; Černá, Marie (advisor) ; Kalina, Tomáš (referee) ; Drda Morávková, Alena (referee)
Autoimmune diseases currently represent the most serious medical issues, mainly due to generally increasing number of patients with these diseases. Their pathogenesis is likely caused by hereditary factors, cellular and humoral interactions influenced by external environmental factors, whose knowledge is important both for diagnostics and therapy, and for theoretical immunology. The aim was to examine the correlation between genetic alterations and the production of autoantibodies, clinical manifestation of the disease; to identify joint autoantigens, further to demonstrate development of leukemic cells from originally autoreactive B cells, the role of B lymphocytes in the pathogenesis of the disease and finally to develop a method for detection of B cells recognizing a defined autoantigen. Several predisposing polymorphisms were revealed using genetic analysis however, they were not exclusively associated either to clinical forms of the disease or usable as prediction markers. In addition, the frequency of alleles IL-1RN * 2 and PD3.1 showed ethno- geographical differences and a critical role of size of sample cohorts in assessing of significance of particular polymorphism was demonstrated in GWA studies. The combination of examination of anti-CCP and IgM RF was found as the best and most...
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Importance of trombophilic mutations in clinical genetic.
Vavrušková, Klára ; Kuklík, Miloslav (advisor) ; Černá, Marie (referee)
Trombophilia means an increased disposition to creation of trombs. Health complication incurred as a consequence of hypercoagulation can be very serious. When a trombophilic mutation is found at a patient, it brings necassity of thrombosis - control in risk situations (e.g. pregnancy, operation) for the rest of the patient's life. There were filed 300 people (206 women and 94 men) with trombophilic mutations into my study of clinical signification of trombophilic mutations. These people were examinated in years 2008 - 2010. Most of positive medical findings - 266 people, were recorded in the area of MTHFR (C677T i A1298C) mutations. There were less findings in the field of FV Leiden and FII prothrombin mutations. Multipath trombophilic mutations were found at 99 patients. I accordance with foreign literature, our results advert to clinical consequences of trombophilic mutations like: repeated spontanious aborts, cerebrovascular akcident (CA), ischaemic heart disease (IHD), thrombosis, flebothrombosis, pulmonary embolism, varicose veins, aseptic necrosis of hip bone, arterial sclerosis and aortic stenosis. Mutations MTHFR C677T and MTHFR A1298C we found mainly at patients with CA, IM and IHD. Leiden mutation was most often found at patients with thrombosis, flebothrombosis and pulmonary embolism. We...
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