|
|
Study of the effect of mesenchymal stem cells in combination with immunosuppressive therapy on inflammatory response in in vivo model
Jabůrek, Filip ; Krulová, Magdaléna (advisor) ; Brdička, Tomáš (referee)
Immunosuppressive drugs have been used for many years for the treatment of autoimmune diseases and post-transplantation treatment. While these drugs have a lot of advantages, they also show several undesirable side effects. The most common side effects are higher blood pressure, lowered renal function and susceptibility to infections. Therefore, in recent years there has been a demand for other medical approaches that do not exhibit the above-mentioned adverse effects. Among one of the newly tested approaches is the application of mesenchymal stem cells (MSCs), which possess several advantages such as immunomodulatory abilities, safety and relatively easy isolation, however, stem cell use alone has not yet provided sufficiently strong immunomodulation. Only a small part of research of MSCs is focused on their use in the combination with immunosuppressive therapy. Therefore, in my thesis I focused on the model which allows to reduce the dose of immunosuppressive drugs in the combination with MSCs. Combined therapy is more advantageous than both monotherapies thanks to lower dosages of these drugs used. It enables to decrease negative side effects of immunosuppressive drugs, when combined with MSCs to provide sufficient immunomodulation in comparison to classical therapy. The aim of my work was to...
|
|
|
Alternative methods for visualization of pancreatic islets.
Gálisová, Andrea ; Jirák, Daniel (advisor) ; Krššák, Martin (referee) ; Kratochvílová, Simona (referee)
Transplantation of pancreatic islets (PIs) represents an alternative treatment for type 1 diabetes mellitus. Post-transplant monitoring of islets by a reliable imaging method may contribute to the improvement of the transplantation outcome. In this thesis, novel visualization approaches for PIs were tested using magnetic resonance (MR) and optical imaging on phantoms and experimental animals, including Chemical Exchange Saturation Transfer (CEST) MR, fluorine (19 F) MR, bioluminescence and fluorescence imaging. MR imaging based on frequency-selective method CEST was performed on islets labeled with Eu-/Yb-based chelates. Labeled islets possessed low MR signal in phantoms, what would have been unsatisfactory for in vivo applications. Moreover, viability and function of labeled islets was impaired reflecting limited applicability of these agents for islet labeling and visualization. Genetically modified bioluminescent islets showed suitable properties for longitudinal tracking of their post-transplant fate at an artificial transplant site - subcutaneously implanted polymeric scaffolds. Using multimodal imaging (MR and bioluminescence), the optimal timing for transplantation of islets into the scaffolds was assessed in diabetic rats. Islets transplanted into scaffolds using the optimized timing scheme...
|
|
|
Study of differential potential of spermatogonial stem cells via transplantation in vertebrates
Kodedová, Barbora ; Krylov, Vladimír (advisor) ; Pšenička, Martin (referee)
Spermatogonia, or spermatogonial stem cells are necessary to maintain male fertility. In the complex process of ongoing spermatogenesis in the testes these pluripotent stem cells proliferate and differentiate into sperm cells. In 1994 the first spermatogonial transplantation technique was described in rodents to allow the study of male germ cells. The following series of studies of intra- and inter-species transmission of testicular tissue revealed the regenerative capacity of transplanted spermatogonial stem cells and their possible usage. Recently, spermatogonia transplantation systems are developed in many vertebrates making it possible to study the development of sperm as well as artificial production of male and female gametes derived from germ cell donors. The differentiation potential of spermatogonial stem cells enabled the creation of transgenic organisms by genetic manipulation of isolated spermatogonia and subsequent transplantation into a suitable recipient. Spermatogonial transplantation in the future may find their application in regenerative medicine, the treatment of disorders of spermatogenesis, or serve to preservation of genetic stock of endangered species.
|
|
|
Huntington's disease modeling and stem cell therapy in spinal cord disorders and injury
Hruška-Plocháň, Marián ; Motlík, Jan (advisor) ; Bjarkam, Carsten (referee) ; Roth, Jan (referee)
Neurological disorders affect more than 14% of the population worldwide and together with traumatic brain and spinal cord injuries represent major health, public and economic burden of the society. Incidence of inherited and idiopathic neurodegenerative disorders and acute CNS injuries is growing globally while neuroscience society is being challenged by numerous unanswered questions. Therefore, research of the CNS disorders is essential. Since animal models of the CNS diseases and injuries represent the key step in the conversion of the basic research to the clinics, we focused our work on generation of new animal models and on their use in pre-clinical research. We generated and characterized transgenic minipig model of Huntington's disease (HD) which represents the only successful establishment of a transgenic model of HD in minipig which should be valuable for testing of long term safety of HD therapeutics. Next, we crossed the well characterized R6/2 mouse HD model with the gad mouse model which lacks the expression of UCHL1 which led to results that support the theory of "protective" role of mutant huntingtin aggregates and suggest that UCHL1 function(s) may be affected in HD disturbing certain branches of Ubiquitin Proteasome System. Traumatic spinal cord injury and Amyotrophic Lateral...
|
|
|
Case Study of Physical therapy treatment of a patient after a combined pancreas and kidney transplantation
Lonek, Jan ; Novotná, Irena (advisor) ; Charvát, Robert (referee)
Title: Case study of physical therapy treatment of a patient after a combined pancreas and kidney transplantation Objectives: The aim of this thesis is to elaborate on case studies of the patient after a combined pancreas and kidney transplant from the perspective of a physical therapist. The case report is supplemented by theoretical information on transplantation and the patient's disease, which is diabetes mellitus. Methods: The thesis is divided into a general and a practical part. Based on the scientific literature, the general section summarizes basic knowledge about diabetes mellitus, simultaneous pancreas-kidney transplantation, and a subsequent physical therapy treatment. The practical part of the thesis describes the process of rehabilitation that the patient underwent during my continuous professional practice at the Institute for Clinical and Experimental Medicine from January 22 to February 16, 2018. Results: During therapy sessions, the patient's overall physical condition and independence were rehabilitated, which were the main goals of the treatment plan. Furthermore, an elimination of reflex changes in soft tissue was achieved, especially in the abdominal area. Keywords: diabetes mellitus, transplantation, pancreas, kidney, physical therapy
|
|
|
ELISpot methodology and prediction of acute rejection after renal transplantation.
Rybáková, Kateřina ; Slavčev, Antonij (advisor) ; Mrázek, František (referee)
Transplantation is the best therapeutic solution for patients with chronic renal failure. Due to the great advances in immunosuppressive therapy in the last decades, graft and patient survival have improved significantly. On the other hand, immunosuppressive therapy has serious side effects - too strong immunosuppression may lead to infection or malignancies, conversely insufficient immunosuppression may lead to graft rejection. Due to the grave consequences of acute rejection, the main goal of cooperation of clinicians and transplant immunologists is to stratify patients into groups with low, moderate and high risk of rejection based on the evaluation of various immunologic risk factors. There are reports in the literature that the numbers (frequencies) of interferon gamma (IFNγ) producing cells before transplantation may be helpful to identify patients with high risk of acute cellular rejection and to predict long-term survival of the graft. In this retrospective study we determined the pre-transplant frequencies of activated donor specific T lymphocytes producing IFNγ after short stimulation (24 hrs) by ELISpot (Enzyme-linked immunosorbent spot assay). The results were correlated with the incidence of acute cellular (ACR) and antibody-mediated (AMR) rejection and with other risk factors. In our...
|
|
|
The role of stem and progenitor cells in regeneration of hematopoietic tissue
Faltusová, Kateřina ; Nečas, Emanuel (advisor) ; Hofer, Michal (referee) ; Filipp, Dominik (referee)
Tissue regeneration is a complex and highly orchestrated process dependent on cells with the potential to restore structures and functions and on controlling factors from the tissue microenvironment. Hematopoietic tissue has a high ability to regenerate, which is attributed to the presence of stem cells, but the regeneration of severely damaged adult tissue is still only partially understood. Hematopoietic tissue provides a unique opportunity to study tissue regeneration due to its well-established steady- state structure and function, easy accessibility, advanced research methods, and well-defined embryonic, fetal, and adult stages of development. Embryonic/fetal liver hematopoiesis and adult hematopoiesis recovering from damage share the needto expand populations of progenitors and stem cells in parallel with increasing production of mature blood cells. We analyzed adult hematopoiesis in mice subjected to a submyeloablative dose (6 Gy) of gamma radiation, in which only a few cells with reconstituting capacity survived. We targeted the period of regeneration characterized by the renewed massive production of mature blood cells and the ongoing expansion of immature hematopoietic cells. Cells from the top of the hematopoietic hierarchy, hematopoietic stem cells, and multipotent progenitors are almost missing...
|
|
|
The role of memory T cells in transplant immunity
Straňavová, Lucia
Memory T cells represent a specific subpopulation of cells formed during the first encounter with antigen. The main role of these cells is to elicit faster and more effective secondary response during reinfections. In transplant immunity, they may affect graft survival directly with donor-specific memory T cells or with cross-reactive virus-specific memory T cells. In this study, we focused on donor-specific and CMV-specific memory/effector T cells. We were interested in the effect of immunosuppressive therapy on the frequency of these cells in periphery. We found that the immunosuppression, prophylaxis and length of dialysis did not significantly affect the number of CMV-reactive cells 6 months after transplantation. We were also interested in the cross-reactivity between CMV and donor antigens, so-called heterologous immunity, which we verified by analyzing the TCR-β repertoire using next- generation sequencing (NGS) in CMV and donor-reactive T cells. Functional cross-reactive T cell clones (shared the same TCR-β sequence) were then found both in the peripheral blood of pre-transplant patients and in the post-transplant graft biopsy. We were also interested if long-term dialysis treatment affects immune memory. Dialysis therapy is often associated with the presence of poorly defined immune system...
|
|
|
Modelovanie ochorenia a štúdium regeneračných procesov v Huntingtonovej chorobe a ALS in vivo
Hruška-Plocháň, Marián
Neurological disorders affect more than 14% of the population worldwide and together with traumatic brain and spinal cord injuries represent major health, public and economic burden of the society. Incidence of inherited and idiopathic neurodegenerative disorders and acute CNS injuries is growing globally while neuroscience society is being challenged by numerous unanswered questions. Therefore, research of the CNS disorders is essential. Since animal models of the CNS diseases and injuries represent the key step in the conversion of the basic research to the clinics, we focused our work on generation of new animal models and on their use in pre-clinical research. We generated and characterized transgenic minipig model of Huntington's disease (HD) which represents the only successful establishment of a transgenic model of HD in minipig which should be valuable for testing of long term safety of HD therapeutics. Next, we crossed the well characterized R6/2 mouse HD model with the gad mouse model which lacks the expression of UCHL1 which led to results that support the theory of "protective" role of mutant huntingtin aggregates and suggest that UCHL1 function(s) may be affected in HD disturbing certain branches of Ubiquitin Proteasome System. Traumatic spinal cord injury and Amyotrophic Lateral...
|
|
|
The meaning of the dead donor rule in current transplantion ethics
Rusinová, Kateřina ; Šimek, Jiří (advisor) ; Kieslichová, Eva (referee) ; Hříbek, Tomáš (referee)
The thesis presents current understanding of the concept of death and criteria for diagno- sis of death in the context of organ donation. We will argue that 1) the dead donor rule should not be the necessary condition for retrieving organs for transplantation and 2) it should be permissible to retrieve organs from patients that are imminently dying (not dead yet), with respect to the principle of autonomy and non-maleficence. We will first present the impossibility and current inconsistencies in determining the exact "moment of death" and we will then demonstrate that current organ donors do not fulfill biological criteria for death and that the dead donor rule is not respected in clinical practice. We suggest that in the context of recent major technological advances in the field of critical care medicine the dead donor rule becomes irrelevant and does not contribute to the transplantation ethics. The legal concept of death and the biological phenomenon of death become more and more distant. We argue that declaring death is not necessary for ethically justified policy in transplantation. Both the societal trust and the protection of vulnerable individuals can be ensured by different ethical principles (i.e. the principle of autonomy and the principle of non- maleficence). The sound ethical...
|
guest ::
