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Evaluation of perfusate solution after ex vivo lung perfusion
Dobrovolná, Terezie ; Enghuber, Florian (referee) ; Paštěka, Richard (advisor)
The aim of this master’s thesis is to introduce lung perfusion procedures and determine the parameters that are connected to the state of the lungs after perfusion and during preservation. In the perfusion process and for storing the lung, the phosphate-buffered saline (PBS) solution is used. The work focuses mainly on the parameters that can be measured and evaluated in the perfusate solution during the perfusion and preservation of the lungs such as pH, total dissolved solids, and proteins. These factors can be measured according to several procedures that are based on different principles. The mentioned process has been selected and implemented with regard to availability, feasibility, and appropriateness given. The pH and TDS parameters have been acquired by electrodes. Therefore, sensor calibration is an integral part of the measure ment. The concentration of the proteins was measured with the Bradford assay where it is important to follow the established protocol. Materials for that were present and provided in the Tissue engineering laboratory. Another intention is to assess the state of the lung tissue, meaning how the lungs gradually change their appearance, weight, and mechanical properties over time. The weight is obtained with the calibrated load cell sensor. For this sensor, a platform had to be assembled in order to function correctly and with great accuracy. From the field of mechanical properties of the lungs, the compliance and PV loops have been selected and presented. Five lungs were used for measurement. The appearance of the lungs changed over time and the necrosis was progressing. The lung viability was preserved with the PBS solution with balanced pH. The value of pH was maintained around pH 7. The concentration of the total dis solved solids (TDS) was determined and similar values were obtained in all lungs because the same solution was used. The TDS fluctuated slowly same as pH because they are correlated. The weight of the lungs decreased with passage of the time as expected. Protein concentration increased over time. That can indicate the inflammatory processes in the lung tissue or lung injury. The surface temperature was also obtained with a laser thermometer and decreasing values were noted with increasing storage time. Static compliance had unexpected charac teristics in some of the lungs. This was probably influenced by leakage due to an unsecured trachea. The last parameter was PV loops. The shape of the PV loops looked good until 48h in the majority of the loops. Then, the shape flatted and the shortage of curves appeared due to shortness of breath.
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Development of an ex vivo lung perfusion system focusing on the preservation of fresh animal lungs for experiments and storage
Mesíková, Klaudia ; Forjan, Mathias (referee) ; Paštěka, Richard (advisor)
A mechanical combined lung model is a type of model used in human breathing simulation. The biggest currency of the model is a high similarity with the human lungs. In order to work with the animal lungs for a longer time and so follow the principles of the 3Rs, a perfusion system is involved in the procedure. The perfusion system filled with a chosen perfusate solution is responsible to prolong the period in which the animal lungs are viable for experiments and storage in the ex-vivo environment. The development of the properly functioning perfusion system is based on the several components included in the process. Choosing the right solution for the perfusion of the inner environment of the lungs is one of the most important things that need to be taken into account. The roller pump is considered the drive motor of the system. Pressure and flow sensors are responsible for monitoring the process parameters that could describe the functionality and the ability to preserve the animal lungs in the ex-vivo environment. The validation of the developed system by using the fresh animal lungs is a part of the thesis as well as the checking procedure of the solution’s influence with the time of the storage. The perfusion system was successfully created and tested. The pressure and flow parameters gained during the measurement were compared while using the saline solution, the Ringer’s solution, and Histofix in the system. The compliance parameter of the lungs were been monitored during the perfusion as well as during the storage with the aim to determine the behaviour of the preserved lungs with the time and the impact of the chosen solution on it. Compliance initially decreased and then stabilized at a certain value throughout the storage period. For the perfusion with the saline and Ringer’s solution, it dropped by one-third. For Histofix preservation, the drop was by half of the initial compliance. The preservation time without the presence of the tissue necrosis was 120 hours using the Saline solution, 240 hours using the Ringer’s solution, and at least 268 hours using Histofix. The perfusion system could further be used in medical research and make a positive aspect in terms of less consumption of the animal organs for experimental purposes in various fields of the research. For future research, the improvement of the perfusion system and solution composition to ensure even longer preservation is welcomed.
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Antimicrobial activity of selected natural extracts
Šandová, Lucie ; Veselá, Mária (referee) ; Zemanová, Jana (advisor)
The aim of the bachelor thesis was to determine antimicrobial activity of selected natural extracts, specifically three model extracts made from purple coneflower (Echinacea purpurea) and four commercial extracts – true lavender (Lavandula angustifolia), lemon balm (Melissa oficinalis), echinacea herbal drops (Echinacea purpurea) and echinacea extract (Echinacea purpurea) with ginger, against selected microorganisms. Theoretical part is focused on description of bioactive substances that are found in plants. It also describes extraction methods used for obtaining extracts from plant materials, methods for determining antimicrobial activity and then the use of natural extracts in food and cosmetic industry. In experimental part, antimicrobial effect of the extracts was tested against gram-negative bacteria Escherichia coli, gram-positive bacteria Bacillus cereus and yeast Candida glabrata. For determination of the antimicrobial activity, two methods (hole method and disc diffusion method) were used and compared. The highest antimicrobial activity against selected microorganisms was showed by commercial extract of lemon balm, which contains lot of geraniol and citroneloll. Significant antimicrobial activity had also been shown by commercial extract of true lavender. On the contrary, extracts from Echinacea had not shown any antimicrobial activity against selected microorganisms.
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Způsoby marinování a nakládání ryb
Fichtnerová, Lenka
The bachelor thesis on the topic "Methods of marinating and pickling fish" deals with the characteristics of individual freshwater and marine fish species, which are the most used in the food industry, their nutritional value, and nutritional properties. The introductory part focuses on the description of specific fish species, the different components of fish meat and their impact on human health. Furthermore, special hygienic rules for handling fishery products were described, which are determined by both Czech legislation and European Union regulations. Subsequently, the various methods of fish processing that ensure the extension of the shelf life of fish meat are explained in detail. This is followed by a description of the various methods, including marinating, pickling, and preserving. Finally, the sub-processes are evaluated in terms of the processing requirements, the raw materials used and the shelf life of the products.
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Evaluation of perfusate solution after ex vivo lung perfusion
Dobrovolná, Terezie ; Enghuber, Florian (referee) ; Paštěka, Richard (advisor)
The aim of this master’s thesis is to introduce lung perfusion procedures and determine the parameters that are connected to the state of the lungs after perfusion and during preservation. In the perfusion process and for storing the lung, the phosphate-buffered saline (PBS) solution is used. The work focuses mainly on the parameters that can be measured and evaluated in the perfusate solution during the perfusion and preservation of the lungs such as pH, total dissolved solids, and proteins. These factors can be measured according to several procedures that are based on different principles. The mentioned process has been selected and implemented with regard to availability, feasibility, and appropriateness given. The pH and TDS parameters have been acquired by electrodes. Therefore, sensor calibration is an integral part of the measure ment. The concentration of the proteins was measured with the Bradford assay where it is important to follow the established protocol. Materials for that were present and provided in the Tissue engineering laboratory. Another intention is to assess the state of the lung tissue, meaning how the lungs gradually change their appearance, weight, and mechanical properties over time. The weight is obtained with the calibrated load cell sensor. For this sensor, a platform had to be assembled in order to function correctly and with great accuracy. From the field of mechanical properties of the lungs, the compliance and PV loops have been selected and presented. Five lungs were used for measurement. The appearance of the lungs changed over time and the necrosis was progressing. The lung viability was preserved with the PBS solution with balanced pH. The value of pH was maintained around pH 7. The concentration of the total dis solved solids (TDS) was determined and similar values were obtained in all lungs because the same solution was used. The TDS fluctuated slowly same as pH because they are correlated. The weight of the lungs decreased with passage of the time as expected. Protein concentration increased over time. That can indicate the inflammatory processes in the lung tissue or lung injury. The surface temperature was also obtained with a laser thermometer and decreasing values were noted with increasing storage time. Static compliance had unexpected charac teristics in some of the lungs. This was probably influenced by leakage due to an unsecured trachea. The last parameter was PV loops. The shape of the PV loops looked good until 48h in the majority of the loops. Then, the shape flatted and the shortage of curves appeared due to shortness of breath.
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The comparison of production technology and conservation of bulky fodder in selected farm and recommendation for improvement
NOVÁKOVÁ, Vladimíra
The first part of this thesis deals with the history and description of roughages. Here, harvesting methods, processing principles and procedures and storage options are described. Also in this first part, the conservation of hay and haylage material is described. For haylage and hay, both sensory properties and laboratory evaluation according to the 'Standard 2004' (No 244/2000 Coll.), which is the most commonly used in the Czech Republic, are given. The most commonly used and commonly occurring clover and grasses are also characterised. Hay additives or preparations are divided into groups and microorganisms for preservation that are harmful or beneficial for biomass fermentation are included. This section concludes with a discussion of maize and the impact of roughages on livestock production. The second part of the thesis deals with the actual monitoring of the quality of roughages, mainly hay pit, hay bale, hay fence and hay bale. Mostly this part focuses on sensory monitoring of the properties of these feeds and laboratory analysis. It concludes with various recommendations that could improve the quality and economics of roughages.
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Účinnost vybraných postupů konzervace ryb na zachování morfometrických ukazatelů
VOGALOVÁ, Pavlína
This bachelor's thesis is focused on the topic of fish fixation and preservation. Experiments regarding various preservation methods are covered in many scientific ichthyological texts. Unfortunately, most of these works are exploring only one preservation method, so mostly there isn't the possibility of comparison with different methods using the same species. Aim of this work is introduction to preservation and fixation methods, that affect morphometric parameters of fish. Theoretical part of this thesis are procedures of fixation and preservation. The experiment itself is focusing on effect of preservation over time. Selected morphometric parameters were observed and measured in selected fish species to discover potential changes in these parameters. Measurements were being taken every two weeks for few months.
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