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The role of mitochondrial DNA in reproduction
Svobodová, Mariana ; Daňková, Pavlína (advisor) ; Jelínková, Ladislava (referee)
The bachelor's thesis focuses on the role of mitochondrial DNA (mtDNA) in human reproduction, especially describing the differences in mtDNA dynamics in oogenesis, spermatogenesis and early embryo development. The physiological function of mtDNA is complemented by its pathologies and their effect on fertility and the ability of the embryo to implant successfully. Furthermore, mtDNA is discussed as a possible diagnostic marker in the evaluation of the quality of sperm, eggs and embryos. These findings of these studies are especially significant for assisted reproduction, where the goal is to select the highest quality embryo with the greatest implantation potential. The work also briefly mentions the presence of cell free mtDNA in the mother's blood and its connection with the development of preeclampsia. Key words: mtDNA, mitochondria, reproduction, (in)fertility
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Effect of biological treatment and body constitution on in vitro osteoclastogenesis from peripheral precursors in axial spondylarthritisis
Beránková, Martina ; Daňková, Pavlína (advisor) ; Rossmeislová, Lenka (referee)
Introduction: Biological therapy is becoming a common approach to stop disease progression and suppress symptoms of axial spondyloarthritis, an inflammatory rheumatic disease characterized by bone loss due to dysregulation of bone turnover and increased osteoclast differentiation. Obesity likely affects bone metabolism through multiple mechanisms. It can promote adipocyte differentiation and fat accumulation while inhibiting osteoblast differentiation and bone formation. Additionally, obesity is closely associated with chronic inflammation, which can enhance osteoclast activity and bone resorption. This master's thesis aims to examine the potential influence of excessive adiposity on the osteoclastogenesis process among patients receiving biological therapy for axial spondyloarthritis. Materials and methods: Monocytes isolated from the peripheral blood of patients with axial spondyloarthritis undergoing biological therapy and healthy donors were stimulated in vitro with pooled sera from patients receiving biological treatment, patients with different type of treatment, and healthy controls for a duration of 7-14 days. Osteoclasts were evaluated as multinucleated, TRAP-positive cells, and their numbers were subjected to statistical analysis. Markers of bone metabolism and inflammation were assessed...
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Molecular mechanisms of cfDNA effects
Hříbková, Natálie ; Daňková, Pavlína (advisor) ; Škubica, Patrik (referee)
The biological role of free circulating DNA is currently being investigated mainly in terms of the etiology of autoimmune conditions and cancer. It appears that the role of cfDNA of nuclear or mitochondrial origin in interacting with receptors of the innate immune system is quite complex and cfDNA is thus involved in a variety of molecular mechanisms such as the spread of cancer metastasis, induction of autophagy and triggering of inflammatory processes. Knowledge of the specific pathways in which cfDNA is involved in the development of these pathologies seems to be crucial, especially in terms of selecting and targeting appropriate therapies. This paper provides a framework for the knowledge on the interaction of cfDNA with receptors of the innate immune system, the induction of immunological signaling pathways and the possible consequences that these molecular pathways may have in the body. Key words: cfDNA, TLR9, cGAS/STING, AIM2, autoimmune disease, biomarker, cancer
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The circulating cell-free DNA as a biomarker of civilization diseases
Dyachenko, Yulia ; Daňková, Pavlína (advisor) ; Vaňková, Markéta (referee)
The incidence of civilization diseases in the population is increasing every year due to changing environmental conditions and modern lifestyles. These diseases reduce the quality of life, lead to health complications or even death of patients. This can be prevented by monitoring health status. Circulating free DNA (cfDNA) has the potential to be introduced as a non-invasive biomarker. Possible mechanisms of cfDNA release are apoptosis, necrosis, NETosis, pyroptosis and active secretion. The occurrence of these processes in some civilization diseases allows the use of cfDNA as a biomarker. The aim of this work was to summarize the current knowledge on the possibility of using cfDNA in the diagnosis, monitoring of the development and course of some diseases of civilization, namely obesity, diabetes mellitus, atherosclerosis, hypertension, myocardial infarction and stroke. From the available literature it can be concluded that cfDNA could be used in clinical practice for the purposes described above. Key words: Circulating cell-free DNA (cfDNA), civilization diseases, biomarker, obesity, diabetes mellitus, atherosclerosis, hypertension, myocardial infarction and stroke.
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Interaction between nurses and family members and their view of patients admitted to the anaesthesia-resuscitation department
DAŇKOVÁ, Pavlína
The present Master's thesis examines the topic of anaesthesia-resuscitation ward visits, interactions, and communication, seen from the perspective and views of nurses and family members of the patients. The primary objective was to examine the interaction between nurses and family members of patients admitted to the anaesthesia-resuscitation wards; the second objective was to investigate the nurses' and family members' perceptions of these patients. The author has achieved the goals mentioned above through qualitative research using two methods: semi-structured interviews with family members of patients and questionnaires involving nurses working in the anaesthesiology-resuscitation wards. The research has established the following: nurses keep an open, positive attitude towards supporting the patient's family; nurses' cooperation with the family members does not depend on their shift difficulty; however, the demandingness of the shift can be influenced by the amount of time the nurse spends with the patient's family; and the nurse's view of the patient is not subject to their education or length of work experience. It is hard for families to see their relatives admitted to the anaesthetic and resuscitation ward; such experiences are associated with very negative feelings; moreover, the family members expect the nurse to behave and act in a role-defined manner. Families usually want to be involved in their loved one's care and appreciate when the nurse is helpful, proactive, supportive and does not shun them; on the other hand, the unfamiliar and strange environment of the ward, and their loved one fighting for life, impacts negatively on families. The present thesis has resulted in drafting an information leaflet that nurses can use for the education of anaesthesia-resuscitation ward visits, containing basic information about the ward, general instructions and a description of the patient, accompanied by illustrations.
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Functional crosstalk between human adipose tissue and lymphatic system
Varaliová, Zuzana ; Rossmeislová, Lenka (advisor) ; Daňková, Pavlína (referee) ; Horáková, Olga (referee)
Both adipose tissue (AT) and lymphatic system (LS) are organs with a great influence on whole-body homeostasis and metabolism. Although the first evidence of a link between the AT and LS was provided already in the 19th century when immune clusters called "milk spots" were found in the omental AT, their functional crosstalk remained unexplored and therefore unappreciated for a long time. However, research in recent years has begun to clearly demonstrate the relationship between the two organs, thanks to the development of new tools for studying and visualizing the LS. Nevertheless, many questions about this crosstalk remain unanswered. This doctoral thesis therefore aimed to answer some of them: How do AT and adipocytes change in response to LS dysfunction ranging from worsened lymphatic drainage to chronic lymphedema? What is the trigger of massive AT expansion in lymphedema? Do adipocytes support lymphangiogenesis, and how? Do lymphatic endothelial cells (LECs) affect the metabolic activities of adipocytes? Although our experiments to answer these questions are ongoing, this work demonstrates several new facts. We showed that the lack of effective lymphatic drainage affects lipolytic reactivity in femoral AT and leads to significant differences in the distribution of AT metabolites between the...
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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...
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Detection of extracellular microRNAs in maternal circulation - diagnosis and prognosis of pregnancy related complications
Ondráčková, Markéta ; Hromadníková, Ilona (advisor) ; Daňková, Pavlína (referee)
MicroRNAs (miRNAs) are small noncoding RNAs of length 18 to 25 nucleotides that regulate gene expression posttranscriptionally. Expression of some miRNAs is tissue specific. I assumed that pregnancy induced complications associated with placental insufficiency could be characterized by a unique profil of placental-specific miRNAs in maternal circulation. I measured concentration and gene expression of selected miRNAs (miR-516-5p, miR-517*, miR-518b, miR-520a*, miR-520h, miR-525 and miR-526a) in the plasma of patients with preeclampsia (PE), fetal growth restriction (FGR) and gestational hypertension (GH). The control group consisted of patients with a normal course of pregnancy (FG). I processed 168 plasma samples, the representation of individual diagnosis were as follows: PE 63, FGR 27, GH 23, FG 55. Detection and quantification was carried out by quantitative real-time PCR. I identified three miRNAs with elevated levels in a group of preeclamptic patients: miR-517*, miR-520a* and miR-525. The severity of the PE, which was characterized by a form of the disease (mild or severe PE) and term (before or after the 34th week of pregnancy), did not have a statistically significant effect on the levels of miRNAs. More than a quarter of patients had PE superimposed on previous hypertension. Previous history of...
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