|
|
Preparation and characterization of enzyme-containing wound dressings
Jurová, Bronislava ; Szotkowski, Martin (referee) ; Skoumalová, Petra (advisor)
This diploma thesis is focused on the study and characterization of nanofiber wound dressings from natural biopolymers. Proteolytic enzymes, specifically collagenase and bromelain, were incorporated into these nanofiber covers. The theoretical part deals with a brief description of skin wounds, their healing and skin covers. There are also methods for preparing these covers and the methods used to characterize them. The practical part deals with the optimization of the preparation of nanofiber coatings based on polyhydroxybutyrate, gelatin, alginate and chitosan. These nanofibers were then enriched with active substances and their gradual release into the model environment was monitored. Finally, their proteolytic activity was determined for these substances.
|
|
|
Study of Biomolecular Interactions with Surface Plasmon Resonance Biosensors
Šípová, Hana ; Homola, Jiří (advisor) ; Houska, Milan (referee) ; Skládal, Petr (referee)
Surface plasmon resonance (SPR) biosensors represent one of the most advanced sensing technologies for real-time studies of biomolecular interactions. In this thesis, methods for functionalization of SPR substrates were optimized and studied via spectroscopic methods. Effects related to the SPR sensor microfluidic interface on the measured biomolecular interactions were analyzed, and furthermore, means to decrease mass-transport limitations were proposed. Several SPR-based assays regarding the detection of nucleic acids were developed, which allow for the detection of physiologically relevant concentrations of nucleic acids as well as point mutations in a nucleic acid sequence. Assays for the determination of the enzymatic activity of HIV integrase and ribonuclease H were developed. These assays can be employed for the design and synthesis of molecules that function either as antiviral drugs or as gene-regulating agents.
|
|
|
Activity of hydrolytic enzymes in process of prodution of biogas
Kolouchová, Kristýna ; Tesařová, Eva (advisor) ; Kopečný, Jan (referee)
In this work, the activities of hydrolytic enzymes, functioning in the hydrolytic stage of the production of biogas, have been determined. The activities of -amylase, -xylanase, and -glucosidase have been measured in five samples collected from the following steps of biogas production:. cow dung from the adjacent cowshed, cow manure from the homogenization tank for dosing biogas plant, digester sludge from the outside biogas fermenter, degister sludge from the inside biogas fermenter, and sludge of digestate from digestate tank. Samples for measurement of enzyme activities were provided by agricultural biogas station AGRI FAIR in Deštná, which uses for biogas production cow manure and corn silage The activities of hydrolytic enzymes in the production of biogas are crucial, because degradability of feed material is the limiting factor. Activity of a amylase had highest value in cow dung and cow manure due to food containing high levels of starch. In subsequent phases falls evenly. In the indoor digester and digestate is already depleted substrate and the activity is at zero level. Its activity is almost three times higher than - glycosidase in cow dung and cow manure, but in the last three stages of -glycosidase activity exceeds amylase and its value is nearly stable. -xylanase has...
|
|
|
Cold adaptation mechanisms in microorganisms
Dřízhalová, Marie ; Nedbalová, Linda (advisor) ; Kvíderová, Jana (referee)
On our planet there are vast areas, where the average temperature does not exceed 5 řC. They are also many places, where the temperature often drops below zero. Not every living organism is able to live in these conditions. The ability to survive in cold region and succefully colonize, such presumably adverse environment requires particular evolutionary adaptation. Adaptation to cold environments has evolved independently in different phylogenetic groups. In low temperature environments, organisms has to adapt in a complex way to be able to permanently inhabit these habitats. Since the analysis of cold adaptation in all organisms is a very complex topic, this thesis is focused on microorganisms. Within this group, it is possible to find certain, which are repeated also in unrelated taxa. The general mechanisms include changes in membrane composition, shifts of enzyme temperature optima, production of compounds that prevent the negative effects of low temperature or the formation of resting stages. This thesis summarizes the mechanisms of cold adaptation which have evolved in microorganisms as response to a long-term or temporary survival at low temperatures. Their potential biotechnological applications are also mentioned. keywords: psychrophilic, psychrotolerant, cold adaptation, enzymes,...
|
|
|
The role of extracellular matrix in mesenchymal stem cell biology
Šimková, Zuzana ; Chudíčková, Milada (advisor) ; Hubálek Kalbáčová, Marie (referee)
The aim of this bachelor thesis is to describe the mutual relations of mesenchymal stem cells and extracellular matrix, their communication and to point out their significance in living body. First part is focused on description of extracellular matrix, its components and characteristics. The interaction between extracellular matrix and surrounding cells is using both receptors and enzymes. Second part addresses the description of mesenchymal stem cells which are crucial producers of extracellular matrix. The matrix is also decisive part of stem cells niche which envelopes them, protects them and decides their future. The last section demonstrates possible utilizations of differentiation potential and paracrine secretion of MSC in therapy and identifies some of the risks bound to it.
|
|
|
Proteolytic Enzymes of Vegetative Forms and Spores of the Bacterium Paenibacillus larvae
Hrabák, Jaroslav ; Martínek, Karel (advisor) ; Kotyza, Jaromír (referee) ; Titěra, Dalibor (referee)
Due to the high resistance of the spores, the bacterium Paenibacillus larvae is the most dangerous bacterial pathogen of the honey bee (Apis mellifera). Thanks to its biological properties and restricted pathogenicity, this bacterium can be used as a model organism to study gram positive sporulating aerobic rods. This work is focused on completing information about secreted proteases of this bacterium and in a study of proteases bound in a spore structure. MYPGP medium was used for the cultivation of P. larvae. In this medium, lysis of the culture was shown after 40 hours of cultivation. The pH of the medium decreased below 6.4 by lysis. The induction of temperate bacteriophage BLA was detected as a causative agent of this lysis. A new sporulation medium called HCBB agar was proposed for the sporulation of P. larvae. In comparison with HCBB agar with MYPGP agar by 31 strains of P. larvae stored in our collection, HCBB agar was evaluated as an appropriate sporulation medium with a median of sporulatin 4.2 ' 106 spores per cm2 in aerobic conditions and 5.65 ' 106 spores per cm2 in aerobic conditions with 10 % CO2. For purification of the secreted proteases, a one-day culture incubated at room temperature was used. Optimal purification of 87/74 kDa and 42/40 kDa proteases was observed after application of this...
|
|
|
New possibilities in wound healing
Nováková, Laura ; Szotkowski, Martin (referee) ; Skoumalová, Petra (advisor)
The diploma thesis is focused on the study of fibrous wound dressings prepared by electrospinning method from natural biopolymers. Three active ingredients were added to the dressings: ampicillin, ibuprofen and collagenase, which are responsible for relieving pain, reducing the risk of infection and selectively removing necrotic tissue in the wound. The theoretical part describes the therapeutic dressings currently available on the market and the most common methods of nanofiber production. The experimental part evaluates the optimization of the preparation of gelatin, alginate and chitosan fibrous wound dressings, which were subsequently enriched with active substances and their gradual release into the model environment was determined spectrophotometrically. Antimicrobial effects against E.coli and S. epidermidis strains andantifungal activity against C. glabrata yeast were monitored. Finally, two cytotoxicity tests on the human keratinocyte cell line HaCaT confirmed the safety of the prepared products, which can serve as bioactive skin dressings in the future.
|
|
|
Isolation and characterization of natural active components and their application in food supplements
Šmídová, Veronika ; Němcová, Andrea (referee) ; Skoumalová, Petra (advisor)
The diploma thesis is focused on the characterisation and isolation of naturally occurred active compounds, especially on lipophilic compounds. The next part of the thesis deals with enzymes and their encapsulation into alginate particles to suggest new enzymatic dietary supplement for children with optimal nutrient composition. The theoretical part is focused on the issue of child nutrition, accordingly the necessary nutrients that children need to grow without any health difficulties. It is also focused on the availability of enzymatic food supplements for children. Finally, the theoretical part deals with the lack of chosen natural substances and their effects in connection with cancer. In the experimental part, four types of oils obtained from seeds were characterized. The oils were obtained by two methods: cold pressing with a Yoda kitchen press and extraction in an organic solvent with Soxtherm. Seeds were linseed, sesame, pumpkin, and cumin. With these oils were characterized natural active substances, polyphenols, flavonoids, antioxidants, carotenoids, chlorophylls, and fatty acids. Liposomes were also prepared from selected extracts, in which an effect on intestinal tumour cells were observed. Furthermore, some enzymes were selected, and they were encapsulated into alginate particles with average size 450 µm. In these particles was specified encapsulation efficiency as well as the proteolytic activity after testing in digest juices. The last part of this thesis was focused on the optimization of a complete enzymatic food supplement with addition of alginate particles as enzyme carriers. Samples of these supplements were finally subjected to a sensory analysis.
|
|
|
Antimicrobial materials based on nanostructures with enzymes
Jurová, Bronislava ; Vysoká, Marie (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis is focused on the preparation and characterization of antimicrobial gels and coatings containing antimicrobial components and enzymes. The theoretical part deals with a brief description of antimicrobial substances, their structure and mainly enzymes with antimicrobial effect. The methods used in the testing are also listed here. The practical part deals with testing the antimicrobial effects of extracts from cinnamon and cloves, clove essential oil, lysozyme and bromelain against Micrococcus luteus, Serratia marcescens and Candida glabrata. Various types of extracts were prepared from the selected spices, namely aqueous and ethanol extracts in different percentages. The individual extracts were characterized and, according to the total proportion of phenolic substances, the best ones were selected for encapsulation in liposome particles and gels. Extracts and enzymes were also added to the hydrogel and non-fibrous covers. A solution of sodium alginate and chitosan was used to prepare non-fibrous covers. A broth dilution method was used to test the antimicrobial effects on the given microorganisms. Furthermore, nanofibers based on PHB were prepared using the forcespinning method. These nanofibers were enriched with clove essential oil and their antioxidant activity was monitored.
|
|
|
Preparation and characterization of wound dressings
Dzurická, Lucia ; Márová, Ivana (referee) ; Skoumalová, Petra (advisor)
The diploma thesis if focused on the study of bioactive hydrogél and nanofiber wound dressings composed of natural biopolymers, which were functionalized by active compounds in the form of analgesic, antibiotics and enzymes. Hydrogél wound dressings were constituted from alginate and chitosan and nanofibers were created from polyhydroxybutyrate. The following 7 active compounds were selected to be added to the wound dressings: ampicillin, streptomycin, ibuprofen, papain, bromelain, collagenase and trypsin. In the theoretical part the structure of the skin and types of wound injuries were described. This part also talks about types of wound dressing and their applications, as well as treatment of skin wounds using enzymes and compounds with analgesic and antimicrobial properties. In addition, this section describes safety assays, in particular cytotoxicity assays on human cells. At the beginning of the experimental part, the process of preparation of hydrogél wound dressing was optimised. Subsequently, the dressings were enriched with active compounds and the rate of gradual releasing of the substances into model environment was monitored. In the case of enzymes, their proteolytic activity was also tested after their incorporation to the wound dressings. Furthermore, the prepared bioactive wound dressings were analyzed for possible cytotoxic effect on human keratinocytes. Finally, the wound dressing with combined content of active substances was created and also characterized for the rate of substance release, proteolytic activity and cytotoxicity. Antimicrobial activity of this wound dressings, against two selected strains of microorganisms: Escherichia coli and Staphylococcus epidermidis, was also evaluated.
|
guest ::
