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Role of peripheral blood monocytes and innate immunity in diabetes
Zinková, Alžběta ; Daňková, Pavlína (advisor) ; Novota, Peter (referee)
Introduction: Diabetes mellitus is a polygenic disease and its development is influenced to some extent by environmental factors as well. Innate immunity triggers nonspecifically first defense reactions after penetration of the pathogen into the body, while overstimulation components of innate immunity may give rise to autoimmune diseases, including diabetes type 1. The components of innate immunity are, among others, Toll-like receptors (TLRs) belonging to a group of the structures recognizing preserved molecular structures characteristic of pathogens. Toll-like receptors are abundantly expressed by monocytes which produce prolactin (PRL) having an immunostimulatory function. To clarify the role of innate immunity in the pathogenesis of diabetes, we focused on the expression of mRNA and protein expression of TLR2 and TLR4. The expression of PRL was studied only at the level of mRNA. Monocytes were separated by flow cytometry into classical (CD14++) and nonclassical (CD14+). We monitored their percentages and the degree of expression of CD14 antigen on their surface.The operational objective of this dissertation was to optimize the stimulation of monocytes for the planned study of the function of non-pituitary prolactin in vitro and determine the appropriateness of the use of healthy donors' buffy...
Comparison of the determination of hormones (Follicle stimulating hormone, Luteinizing hormone, Prolactin, Testosterone, Progesteron) by two analytical systems. Converting accredited method and its verification.
Kucejová, Soňa ; Martínková, Markéta (advisor) ; Mrízová, Iveta (referee)
Analytical system ARCHITECT i2000SR was verified according to requirements of ÚLBLD VFN and 1. LF UK laboratory in Prague. Repeatability, intermediate precision, and measurement uncertainty were determined as performance parameters for verification of analytical assays for testosterone, progesterone, luteinizing hormone, follicule stimulating hormone and prolactin. Results of Lyphochek control samples, which were measured, were consistent with values given by manufacture. Repeatability: coefficients of variation for testosterone Lyphochek 1 6,81%, for Lyphochek 3 6,40%, progesterone 2,4% and 1,8%, luteinizing hormone 5,38% and 1,89%, follicle stimulating hormone 5,12% and 3,24% prolactin 1,45% a 1,83%. Intermediate precision: coefficients of variation for testosterone Lyphochek 1 6,02%, Lyphochek 2 3,60%, Lyphochek 3 3,07%, progesterone 7,9%, 4,9% and 5,8%, luteinizing hormone 4,50%, 5,51% and 5,83%, follicle stimulating hormone 4,00%, 3,72% and 4,87%, prolactin 4,60%, 4,20% and 5,00%. Measurement uncertainty: testosterone 6,02%, progesterone 7,9%, luteinizing hormone 5,83%, follicle stimulating hormone 4,87%, prolactin 5,00%. Analytical System Architect i2000SR was compared with previously used ADVIA Centaur system to find out, whether it is possible to convert the method Centaur Testosterone,...
Role of toll-like receptors and stress hormone prolactin in defects of immune system
Sluková, Veronika ; Daňková, Pavlína (advisor) ; Hušáková, Markéta (referee)
Introduction: Diabetes mellitus is a polygene disease and on its manifestation have influence also enviromental factors. We have studied the role of extrapituitary prolactin (PRL) and toll-like receptors (TLR) 2 and 4 in the etiopathogenesis of autoimmune diabetes. PRL is mainly produced by hypophysis, but in small concentrations also in the periphery, where it participates in the immune reactions. Therefore, we investigated the influence of the levels of monocytic PRL mRNA on the development of diabetes, and also the influence of G allele of the -1149 G/T polymorphism in the extrapituitary promotor, which has already been associated with other autoimmune diseases. TLRs are receptors of the immune cells that recognize patogenes entering into the body. They play an important role in the iniciation of the immune response. We aimed to find out their function in the pathogenesis of the autoimmune diabetes by the detection of their mRNA levels and protein levels expressed on the cell surface of the monocytes. Material and methods: In this study we included 30 T1D and 21 LADA patients. Three control groups consisted of 23 T2D patients, 23 patients with a nondiabetic disease (neDM) and 60 healthy blood donors (TO). Blood samples have been taken from the individuals. From these blood samples we isolated...
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...
Neuroimmune and endocrine correlates of stress response and dissociation in affective disorders
Bízik, Gustáv ; Bob, Petr (advisor) ; Horáček, Jiří (referee) ; Yamamotová, Anna (referee)
Depression and other mental disorders are the leading cause of disability worldwide and their burden has increased considerably over past decades. However, advances in psychopharmacology of psychiatric disorders are not in measure with this negativ trend. As a result, a large body of researchinpsychiatryandneurosciencestries to furtherourunderstanding of pathophysiologicalmechanismsunderlyingmooddisorders andothermentalillnesses in order to improve the efficacy of current treatments and to identify new therapeutic agents. According to current evidence, stress-related pathways and inflammation processes are directly involved in thedevelopment of depressive disorder andseveral other psychiatric conditions.Thestudy of the effects and consequences of stress exposure requires an interdisciplinary approach,taking into account specific aspects of the "inputs", such as chronic stress and traumatic experiences, and related psychological processes, with the crucial role of dissociation. Following these theoretical findings, the empirical research performed in two cohorts of inpatients with depressive disorder focused on immune and endocrine responses to stress and their relationship to psychopathological symptoms, specifically trauma-related symptoms, psychic and somatoform dissociation and depressive...
Immunogenetic and hormonal markers of predisposition to systemic rheumatic diseases particularly systemic lupus erythematosus
Fojtíková, Markéta ; Pavelka, Karel (advisor) ; Hrnčíř, Zbyněk (referee) ; Rovenský, Jozef (referee)
Fojtikova 2011 INTRODUCTION: Several factors like genetic susceptibility is required for systemic rheumatic diseases development. Immunomodulatory PRL effect supports autoimmunity. AIMS: 1. To detect the immunogenetic background (alleles HLA class I, II and microsatellite polymorphism of the transmembrane part exon 5 of MIC-A gene) of SLE and PsA. 2. To detect PRL serum and synovial fluid with regard to clinical and laboratory RA activity. 3. To find the role of the functional polymorphism -1149G/T SNP PRL of extrapituitary promoter of PRL gene in SLE, RA, PsA, SSc and inflammatory myopathies development. METHODS: Genetic analyses of pateints with SLE (n=156), RA (n=173), PsA (n=100), SSc (n=75), PM (n=47) a DM (n=68) and 123 healthy individuals: PCR-SSP (HLA clase I and II), PCR-fragment analysis (MIC-A) a PCR-RFLP (-1149 G/T SNP PRL). In 29 RA a 26 OA PRL serum and synovial fluid concentrations were detected using immunoradiometric assay. RESULTS: 1. The allele HLA-DRB1*03 (pc=0.008; OR 2.5) and haplotype HLA-DRB1*03-DQB1*0201 (pc <0.001; OR 4.54) were determined as risk immunogenetic markers for SLE in Czech population. In SLE versus controls allele MIC-A5.1 was increased (pc =0.005; OR 1.88). MIC-A5.1 together with HLA-DRB1*03 increases the risk for SLE development, pc <0.000001; OR 9.71....
The hypothalamic-pituitary-gonadal axis and epilepsy: mutual relationships
Čuchalová, Marcela ; Herink, Josef (advisor) ; Semecký, Vladimír (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Science Author: Marcela Čuchalová Supervisor: doc. MUDr. Josef Herink, DrSc. Title of diploma thesis: The hypothalamic - pituitary - gonadal axis and epilepsy: mutual relationships The content of the diploma thesis is an overview of the anatomy and physiology of the hypothalamic-pituitary-gonadal axis (HHG). Further chapters are devoted to the influence of epilepsy on HHG function, the effect of HHG hormones on epileptic activity itself. The effect of anti-epileptics on HHG functions will also be elucidated. The second part of the diploma thesis deals with separate chapters - catamenial epilepsy and epilepsy during pregnancy. Keywords: antiepileptic drugs, gonadotropin, hypothalamic-pituitary-gonadal axis, prolactin, sex hormones, temporal lobe epilepsy.
Neuroimmune and endocrine correlates of stress response and dissociation in affective disorders
Bízik, Gustáv ; Bob, Petr (advisor) ; Horáček, Jiří (referee) ; Yamamotová, Anna (referee)
Depression and other mental disorders are the leading cause of disability worldwide and their burden has increased considerably over past decades. However, advances in psychopharmacology of psychiatric disorders are not in measure with this negativ trend. As a result, a large body of researchinpsychiatryandneurosciencestries to furtherourunderstanding of pathophysiologicalmechanismsunderlyingmooddisorders andothermentalillnesses in order to improve the efficacy of current treatments and to identify new therapeutic agents. According to current evidence, stress-related pathways and inflammation processes are directly involved in thedevelopment of depressive disorder andseveral other psychiatric conditions.Thestudy of the effects and consequences of stress exposure requires an interdisciplinary approach,taking into account specific aspects of the "inputs", such as chronic stress and traumatic experiences, and related psychological processes, with the crucial role of dissociation. Following these theoretical findings, the empirical research performed in two cohorts of inpatients with depressive disorder focused on immune and endocrine responses to stress and their relationship to psychopathological symptoms, specifically trauma-related symptoms, psychic and somatoform dissociation and depressive...
Comparison of the determination of hormones (Follicle stimulating hormone, Luteinizing hormone, Prolactin, Testosterone, Progesteron) by two analytical systems. Converting accredited method and its verification.
Kucejová, Soňa ; Martínková, Markéta (advisor) ; Mrízová, Iveta (referee)
Analytical system ARCHITECT i2000SR was verified according to requirements of ÚLBLD VFN and 1. LF UK laboratory in Prague. Repeatability, intermediate precision, and measurement uncertainty were determined as performance parameters for verification of analytical assays for testosterone, progesterone, luteinizing hormone, follicule stimulating hormone and prolactin. Results of Lyphochek control samples, which were measured, were consistent with values given by manufacture. Repeatability: coefficients of variation for testosterone Lyphochek 1 6,81%, for Lyphochek 3 6,40%, progesterone 2,4% and 1,8%, luteinizing hormone 5,38% and 1,89%, follicle stimulating hormone 5,12% and 3,24% prolactin 1,45% a 1,83%. Intermediate precision: coefficients of variation for testosterone Lyphochek 1 6,02%, Lyphochek 2 3,60%, Lyphochek 3 3,07%, progesterone 7,9%, 4,9% and 5,8%, luteinizing hormone 4,50%, 5,51% and 5,83%, follicle stimulating hormone 4,00%, 3,72% and 4,87%, prolactin 4,60%, 4,20% and 5,00%. Measurement uncertainty: testosterone 6,02%, progesterone 7,9%, luteinizing hormone 5,83%, follicle stimulating hormone 4,87%, prolactin 5,00%. Analytical System Architect i2000SR was compared with previously used ADVIA Centaur system to find out, whether it is possible to convert the method Centaur Testosterone,...
Role of peripheral blood monocytes and innate immunity in diabetes
Zinková, Alžběta ; Daňková, Pavlína (advisor) ; Novota, Peter (referee)
Introduction: Diabetes mellitus is a polygenic disease and its development is influenced to some extent by environmental factors as well. Innate immunity triggers nonspecifically first defense reactions after penetration of the pathogen into the body, while overstimulation components of innate immunity may give rise to autoimmune diseases, including diabetes type 1. The components of innate immunity are, among others, Toll-like receptors (TLRs) belonging to a group of the structures recognizing preserved molecular structures characteristic of pathogens. Toll-like receptors are abundantly expressed by monocytes which produce prolactin (PRL) having an immunostimulatory function. To clarify the role of innate immunity in the pathogenesis of diabetes, we focused on the expression of mRNA and protein expression of TLR2 and TLR4. The expression of PRL was studied only at the level of mRNA. Monocytes were separated by flow cytometry into classical (CD14++) and nonclassical (CD14+). We monitored their percentages and the degree of expression of CD14 antigen on their surface.The operational objective of this dissertation was to optimize the stimulation of monocytes for the planned study of the function of non-pituitary prolactin in vitro and determine the appropriateness of the use of healthy donors' buffy...

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