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Pharmacokinetics of Intramuscularly Administered Thermoresponsive Polyacrylamides
Groborz, Ondřej
6 Pharmacokinetics of Intramuscularly Administered Thermoresponsive Polyacrylamides Author: Ondřej Groborz Supervisor: doc. Mgr. Martin Hrubý, Ph.D., DSc. Institute of Macromolecular Chemistry, Czech Academy of Sciences Advisers: Ing. Pavel Švec RNDr. Lenka Loukotová, PhD. Abstract Polymer solutions with lower critical solution temperature (LCST) undergo a phase separation when heated above their cloud point temperature (TCP). These thermoresponsive polymers have numerous promising medicinal applications, such as in situ depot-forming radiotherapy (brachytherapy), controlled drug-release, immuno-radiotherapy, injectable thermogelling for tissue engineering and cell culture and magnetic resonance imaging (MRI), among others. Yet, despite extensive research on medicinal applications of thermoresponsive polymers, their fate after their administration remains largely unknown. Thus, in our study, we synthesized and thoroughly characterized four different thermoresponsive polyacrylamides, namely poly(N-(2,2-difluoroethyl)acrylamide), poly(N- isopropylacrylamide), poly(N,N-diethylacrylamide) and poly(N-acryloylpyrrolidine) under physiologically relevant conditions. Subsequently, we determined their biodistribution kinetics in mice and proposed a data-based pharmacological model to describe their in vivo behaviour,...
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Antivirotické účinky biologicky aktivních látek z přírodních zdrojů v kombinaci s imunomodulačními látkami proti viru klíšťové encefalitidy
ČERNÝ, Jan
The aim of this study is focused on substances from the nature as a potentional treatment against tick-born encephalitis virus. The research of this master thesis can be divided into two sections. The first one is about choosen derivatives of adamantane, their cytotoxic effect on neuron cell lines and the determination of their antivirotic properties in vitro. For these purposes was used the combination of MTT assay, plaque titration assay and one-step qRT-PCR. The second part of this work verified the capability of choosen stilbenes to prolong the life of BALB/c mice which were infected by tick-born encephalitis virus.
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Pharmacokinetics of Intramuscularly Administered Thermoresponsive Polyacrylamides
Groborz, Ondřej ; Hrubý, Martin (advisor) ; Matějíček, Pavel (referee)
6 Pharmacokinetics of Intramuscularly Administered Thermoresponsive Polyacrylamides Author: Ondřej Groborz Supervisor: doc. Mgr. Martin Hrubý, Ph.D., DSc. Institute of Macromolecular Chemistry, Czech Academy of Sciences Advisers: Ing. Pavel Švec RNDr. Lenka Loukotová, PhD. Abstract Polymer solutions with lower critical solution temperature (LCST) undergo a phase separation when heated above their cloud point temperature (TCP). These thermoresponsive polymers have numerous promising medicinal applications, such as in situ depot-forming radiotherapy (brachytherapy), controlled drug-release, immuno-radiotherapy, injectable thermogelling for tissue engineering and cell culture and magnetic resonance imaging (MRI), among others. Yet, despite extensive research on medicinal applications of thermoresponsive polymers, their fate after their administration remains largely unknown. Thus, in our study, we synthesized and thoroughly characterized four different thermoresponsive polyacrylamides, namely poly(N-(2,2-difluoroethyl)acrylamide), poly(N- isopropylacrylamide), poly(N,N-diethylacrylamide) and poly(N-acryloylpyrrolidine) under physiologically relevant conditions. Subsequently, we determined their biodistribution kinetics in mice and proposed a data-based pharmacological model to describe their in vivo behaviour,...
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Multipotent mesenchymal stromal cells in orthopedic: Potentiation of bone healing
Stehlík, David ; Trč, Tomáš (advisor) ; Janíček, Pavel (referee) ; Sedmera, David (referee)
The aim of the thesis was development of an innovative treatment of bone defects. Human multipotent mesenchymal stromal cells (MSC) play a crucial role in bone healing. Clinical applications of MSC require large amount of cells, which could be obtained by autologous expansion of MSC harvested from bone marrow. As a first step, the standard protocol of MSC expansion based on αMEM medium and fetal bovine serum (FBS) was used. Experiments replacing FBS by pooled human serum (HS) in the culture medium concluded in patenting of a new MSC cultivation protocol (EU 1999250, CR 301141). This one-step cultivation protocol and xenogeneic protein-free cultivation medium is based on CellGro® for Hematopoietic Cells' Medium, HS, human recombinant growth factors, dexamethasone, insulin and ascorbic acid. The preclinical in vitro and in vivo experiments with MSC from both expansion protocols were carried out. Fibrillar polylactic scaffolds were seeded with MSC, cultured, differentiated and implanted in immunodeficient mice (NOD/LtSz-Rag1-). Bone-like mineralized tissue containing vessels was observed. The MSC cultured according to patented method were classified as Advanced-therapy Medicinal Product and has to fulfil the European Medicines Agency regulations to enter the clinical trials. Nevertheless the use of MSC seems...
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Bioaktivní látky ve vodním prostředí a jejich vliv na ryby - zaměření na cytochromy P450 v rybách
SAKALLI, Sidika
The term "biologically active compounds" covers a wide range of substances originating from natural or synthetic origins. These compounds can enter the aquatic environment through wastewater treatment plants, manufactural discharge or they are naturally present in the aquatic plants or microorganisms. Their adverse effects on fish has been widely studied and accepted. This thesis is focussed on the adverse effect of several bioactive compounds (i.e. pharmaceuticals, phytochemicals, or complex mixture of pollutants) on fish using different experimental design as in vitro, in vivo and in situ. In the first part of this thesis, in vitro effects of pharmaceuticals and phytochemicals or their combinations on fish CYP system were investigated. Moreover, effects of standard carrier solvents used in enzyme activity assays were also investigated. An important finding in this study was the lack of effects of either dexamethasone, quercetin, or indole-3-carbinol on EROD activity; however, when these agents were combined, EROD activity was strongly inhibited. This demonstrates that combination of compounds might exert different effects than single compounds, and the effects of mixture compounds cannot be predicted from the effects of individual compounds. In the second part of the thesis, the effects of chronic exposures of rainbow trout to dexamethasone and clotrimazole were investigated. The results regarding dexamethasone showed similarities with in vitro studies, and both in vitro and in vivo exposure of dexamethasone did not alter the CYP enzyme activities. On the other hand, in vivo exposure of clotrimazole yielded conflicting results with findings from the in vitro studies. Clotrimazole induced EROD activity in chronically exposed rainbow trout, and BFCOD activity showed biphasic pattern in which it was inhibited at environmentally relevant concentrations and induced at high concentrations. Thus, the observed effects suggest that clotrimazole could negatively affect fish CYPs at environmentally relevant concentrations. However, in the environment, the effects of clotrimazole and dexamethasone on fish CYPs system might be modified because of unknown compounds in these mixtures. Therefore, further investigations were done to identify the effects of mixture compounds using an in situ model. The last part of the dissertation addresses the effects of cocktail PPCPs on common carp under natural conditions. In situ studies provide valuable information on both hepatic and intestinal CYP activities. Both EROD and BFCOD activities were affected by the PPCPs that are present in the exposed fish. Moreover, changes in intestinal CYP activities suggest that fish can ingest some of these contaminants through their feed. Therefore, the intestines might be responsible for elimination of some of these pollutants and act as the first barrier of pollutant entry in fish. Despite the extensive studies concerning aquatic pollution, further studies are necessary. Development of new pharmaceuticals, their occurrence in the aquatic environment, and their effects on non-target organisms should be continuously monitored.
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Multipotent mesenchymal stromal cells in orthopedic: Potentiation of bone healing
Stehlík, David ; Trč, Tomáš (advisor) ; Janíček, Pavel (referee) ; Sedmera, David (referee)
The aim of the thesis was development of an innovative treatment of bone defects. Human multipotent mesenchymal stromal cells (MSC) play a crucial role in bone healing. Clinical applications of MSC require large amount of cells, which could be obtained by autologous expansion of MSC harvested from bone marrow. As a first step, the standard protocol of MSC expansion based on αMEM medium and fetal bovine serum (FBS) was used. Experiments replacing FBS by pooled human serum (HS) in the culture medium concluded in patenting of a new MSC cultivation protocol (EU 1999250, CR 301141). This one-step cultivation protocol and xenogeneic protein-free cultivation medium is based on CellGro® for Hematopoietic Cells' Medium, HS, human recombinant growth factors, dexamethasone, insulin and ascorbic acid. The preclinical in vitro and in vivo experiments with MSC from both expansion protocols were carried out. Fibrillar polylactic scaffolds were seeded with MSC, cultured, differentiated and implanted in immunodeficient mice (NOD/LtSz-Rag1-). Bone-like mineralized tissue containing vessels was observed. The MSC cultured according to patented method were classified as Advanced-therapy Medicinal Product and has to fulfil the European Medicines Agency regulations to enter the clinical trials. Nevertheless the use of MSC seems...
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Invasive structures of cancer cells in 3D environment
Lyková, Dominika ; Tolde, Ondřej (advisor) ; Libusová, Lenka (referee)
The ability of cells to migrate through tissue barriers plays an important role in physiological and pathological processes including immune response or invasiveness of cancer cells. The cells generate cytoplasmic protrusions called podosomes and invadopodia, collectively known as invadosomes or podosome-type adhesions (PTA), which are thought to be the key structures of cell invasion, especially of cancer cells during metastasis. Invadosomes are F-actin rich cell-matrix contacts with capability to degrade extracellular matrix components and are observed both in normal cells (such as monocytic cells, endothelial cells and smooth muscle cells) and in cancer cells. This bachelor thesis is focused on those in cancer cells, their initiation, regulation, function and morphology in 3D and in vivo and their requirement for tumor metastasis.
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Glycaemic index of food and its determination
Špiláčková, Aneta ; Kouřimská, Lenka (advisor) ; Monika, Monika (referee)
The purpose of this thesis was to elaborate a literature recherche on a given theme with focus on analytical methods to estimate the glycaemic index of food.
This bachelor thesis is divided into three main chapters. First chapter describes general characteristics of glycaemic index of foods and influencing factors. In this chapter states the influence of high and low glycaemic index foods on health, wellbeing and glycaemia in humans. Subsequent part of this chapter is dealing with many other factors that influence the absolute amount of glucose in blood in 2 hour interval after meal.
Chapter two initiates into conventional and continual methods of glycaemia measurement.It highlights the advantages and limitations of each of these methods.
Third, the most extensive chapter is dealing with methods of glycaemic index assessment. This chapter is broken down into two parts. First part specifies clinical methods that are also called in vivo, and the second part is focusing on in vitro methods. Clinical methods require blood glycaemia measurement in healthy volunteers after consuming a test meal. With this method we are able to reliably state the glycaemic index of certain foods.
The limiting factor of this method is that it is rather time-consuming and it requires higher financial means. These measurements take place under a strict control, according to guidelines and only with the approval of ethical committee.
Analytical methods are based on simulation of human digestion. In vitro methods use hydrolysis index to obtain the glycaemic index. The final values are only an assessment of glycaemic index of given food. Other method that is described is measurement of fast and slowly available glucose and starch in given test foods. It is based on measurement of released glucose from the test food with HPLC method, time incubation and standardized conditions. Last method includes partitioning of a test food, and subsequent HPLC analysis of carbohydrates and sugar alcohols. The HPLC results are processed in artificial neural network to reveal glycaemic index values.
The advantages and disadvantages of each mentioned method are stated in conclusion of this work. I compared and contrasted reliability, elaborateness, budget demands
of analytical methods with clinical assessment methods.
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Záchrana genofondu vybraných starých odrůd růží z areálu Flora Olomouc množením in vitro
Remerová, Pavla
In the thesis "Saving the gene pool of selected varieties of old roses from the area Flora Olomouc using in vitro propagation" are listed procedures for sampling explants and primary cultures foundation varieties of roses Klimentina, Jan Palach, Fortissimo, Pastorale and Julius Fabiancs de Misefa. It was developed a protocol in vitro propagation of roses and selected media and culture conditions for the maintenance of multiplication in gene bank. Varieties Klimentina, Jan Palach, Fortissimo, Pastorale and Julius Fabiancs de Misefa formed its foundation. An integral part of the above results indicate that the formation of roots and convert rooted plants in non-sterile conditions. Finally, we tested the stability of donor plants and plant tissue using SSR primers in the varieties Klimentina, Jan Palach, Fortissimo and Julius Fabiancs de Misefa.
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