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Optimization of recombinant protein expression in HEK293 cell line
Bláha, Jan ; Šácha, Pavel (referee) ; Bezouška, Karel (advisor)
Mammalian cells have become the dominant system for recombinant expression of pharmaceutical proteins. This system is becoming suitable also for structural biology, with the advances in methodology of transient transfection of mammalian cells. This work dealt with optimization of recombinant expression in HEK293T and HEK293-6E cell lines in various media using easily quantifiable markers - secreted alkaline phosphatase (SEAP) and green fluorescent protein (GFP). Emphasis was placed on optimizing key factors behind the creation of transfection complex - the ratio of DNA to polyethyleneimine and the amount of DNA used. The positive influence of histone deacetylases inhibitor valproic acid and also of casein hydrolysate Tryptone N1 was also studied. (The thesis is written in Czech.)
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Therapeutic importance of manual lymphatic drainage for lymphedema of limbs
VOTRUBOVÁ, Eva
The importance of lymphatic drainage and other techniques used to cure lymphedema was known as early as around the year 3,000 BC. In the Czech Republic it became quite common after 1992, when the Czech Society of Lymphology was founded. Since the number of professionals in this area has been increasing and so has the number of patients, which makes the cure of lymphedema a very topical question. The first chapter of the theoretical part of my thesis is devoted to the history of the anatomy of lymphatic system, lymphedema and manual lymphatic drainage. There are also chapters on the definition of lymphedema and its classification according the mechanism of its creation and pathophysiology which triggers this problem. The differences between primary and secondary lymphedema are described in detail. I also sum up the knowledge about the etiology of lymphedema, its clinical picture, its differential diagnosis and stages of development. Apart from surgical, pharmacological and spa treatment I deal with decongestive treatment, which includes instrumental lymphatic drainage, banding, regime measurements, special exercise and manual lymphatic drainage. The chapter on manual lymphatic drainage deals both with effect and indications and contraindications of this method. The basic aim of my thesis is to map the contribution of manual lymphatic drainage to the cure of the lymphedema of limbs with special attention to the span of motion of joints and circumference of the limb before and after the therapy. The method of quantitative research was used for the practical part. The research was done on 3 respondents, who went through the entry examination. It contained the taking of thorough anamnesis including the personal data and family, social, professional etc. anamnesis. I also did a kinesiology analysis, which contained the aspection from the front, from the side and from the back and the values which were of the highest importance to me: circumference of the limbs and goniometric measuring of the span of motion of joints of the afflicted limb in comparison to the healthy one. The first research question is: What is the influence of the manual lymphatic drainage on circumference of the afflicted limb? The second question is: What is the influence of the manual lymphatic drainage on the span of motion of joints of the afflicted limb? During my internship in the rehabilitation centre in Borovany I had consultations with Jana Poláková, BA, who is a graduate of a certified course of manual lymphatic drainage. We discussed the choice of suitable patients, possible therapy and short-term and long-term plans. After every first or second massage I conducted a check measurement with all probands. Al in all conducted 5 measurements with each proband. The aim of my thesis was to map the contribution of the lymphatic drainage to the rehabilitation and its positive influence on the overall improvement of the patient´s health. The research was conducted in the rehabilitation centre in Borovany through an interview, retrospective data analysis, measurements of the circumference of the limbs and the span of motion of joints and observation and examining of the afflicted limbs. Subjectively all the patients find manual lymphatic drainage useful in the treatment of lymphedema. They mention improvements in the visible decrease in circumference of the limbs and the span of motion of joints as well as the overall perception of the limbs, their posture and motoric. Objectively the decrease of centimetres in circumference of the limbs and increase in the span of motion of joints was observed with all the patients.
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Care about immature newborn of HIV positive mother
Marianová, Sandra ; Fajt, Martin (referee) ; Saxlová, Jaroslava (advisor)
This thesis is dedicated to the care of immature newborn HIV positive mothers. Work is conceived as a theoretical - practical. The theoretical part is devoted to HIV infection, their transmission, including transmission from mother to child, diagnosis, treatment options and preventive measures. Although there is enough awareness and prevention programs aimed at preventing transmission and transmission of HIV infection, a large number of HIV positive women diagnosed during pregnancy, resulting in considerable risk to the fetus. Then the paper deals with the care of HIV positive women during pregnancy and during delivery. Another section is dedicated to the care of newborn immature, are mentioned aspects of medical and nursing care for these children. The practical part contains a case report of immature babies who are born HIV positive mother. Case report describes the course of hospitalization for Neonatology Intensive Care Unit. There is comprehensively described for newborn care, focusing on the specifics of treatment of immature children. It also mentioned the proportion of parental care, including their education in the care of these children.
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Virtual construction of social reality through online communication and social networks
Klouček, Viktor ; Rosický, Antonín (advisor) ; Sigmund, Tomáš (referee)
This final paper is centered on influence of the internet in forming of social reality. Main factor in creation of social reality is element of socialization, say communication. The aim of this paper is online communication and socialization developed through chats, discussion groups, blogs, forums summarized by one term -- social networks, which gained very strong position among communication applications available on the internet. I have tried to find out how online communication influence people and how close one can get in creation of social reality in virtual settings in correlation with other members of such settings. Technology of today creates new forms of communication and new ways and connections among members of online communication groups, which differ from the real ones. Distance is no longer an issue and it is completely natural to interact, thus influence somebody and be influence by somebody, from a different geographical, social, economical or political settings. All of these factors directly influence construct of social reality of those who communicate. Study was conducted as a survey. Questionnaire was sent to two hundred people and 107 returned back. More than half of 107 respondents don't know each other, come from various social classes and different areas of Czech Republic. Range of age of respondents is 15-63 years. All of them were given questions aiming at the reality of virtual communication, virtual friendship and bonds between online members. Other questions aim at the authenticity of information and partially influence of commercials in process of decision making. My contribution to this work was mainly creation of questioner, analysis of answers towards conclusions about the influence of online communication on perception and forming of social reality. Analysis that aimed at respondents spending most of their time online brought interesting conclusions in comparison with other groups, which didn't spent as much time online. The paper begins with theoretical summary of reality, its construction, online applications and scientific papers which include their positive and negative impacts. Following there is study introducing respondents, basic questions, hypothesis and then interpretation of answers with previous studies taken in account. Outcome of this work concludes main aspects of interpretations and draw basic view of the future. The questionnaire and more detailed answers about time spent online are in the appendix.
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The effect of local heating on air quality in a small settlement
Andělová, Ludmila
Concentrations of PM10 and Black Smoke (BS) were measured during one winter and two summer periods in small village Albrechtice v Jizerských horách. Major source of aerosol in this village is domestic use of coal and wood burning. 15 minute concentrations of PM10 were measured by DustTrak and 24-h concentrations of PM10 were measured by Harvard impactor. 24-h concentrations of BS were measured too. Concentrations of PM10 and 24-h concentrations of BS were highly correlated during the heating season (0,89). The correlation dropped to 0,68 during the second summer period. Concentrations of PM10 were negatively correlated with average daily temperature during the winter season (-0,41). Positive correlation between PM10 and temperature was found during the first summer period (0,34) and no correlation was found during the second summer period (-0,01). Concentrations of BS were negatively correlated with average daily temperature during all the three seasons. Negative correlation between PM10 and wind velocity was found during the winter season (-0,43). Daily maximums were found mainly between 4 and 12 pm. Comparison of PM10 data from Albrechtice with PM10 data from two urban areas Jablonec nad Nisou and Liberec and one background area Souš showed that average PM10 concentrations in Albrechtice were the...
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Abiotic elicitation of explant culture of Trifolium pratense L.
Fiedlerová, Gabriela ; Siatka, Tomáš (referee) ; Kašparová, Marie (advisor)
Abiotic elicitation of explant culture of Trifolium pratense L. Summary The objective of this dissertation was to observe the effect of the abiotic elicitors of mercuric chloride and cadmium chloride on the production of flavonoids, and also of the effect of the mercuric chloride elicitor on the production of isoflavonoids in the Trifolium pratense L. explant culture (variety DO−8). The cultures were cultivated in the Gamborg nutrient media with the addition of 2,4−dichlorphenoxyacetic acid (2 mg.l-1 ) and 6−benzylaminopurine (2 mg.l-1 ), in the temperature of 25 řC and during a 16-hours light/8-hours dark period. The quantity of flavonoids was determined spectrophotometrically according to The Czech Pharmacopoeia 2005. The quantity of isoflavonoids was determined by the HPLC method. We can see from the results of this dissertation that the concentration 10 μmol of both of the used abiotic elicitors had the greatest positive effect on the production of flavonoids. The largest quantity of flavonoids (0,576 %) was detected during the elicitation of cadmium chloride of a 10-μmol concentration and after 48−hour application when the quantity of flavonoids increased by statistically significant 294 % in relation to the control sample. The quantity of isoflavonoids was determined by the HPLC method during the...
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The Ukrainian family names
Jarošová, Kateřina ; Lendělová, Věra (advisor) ; Savický, Nikolaj (referee)
In my thesis I follow the issue of Ukrainian family names. The study of anthroponymics in itself is still of live reflection due to the fact that both first and family names make it possible to get important data not only from philological point of view (for language structure research) but also from historical and ethnographical ones. Key guiding sources for this essay were: Publication "Sučasni ukrajinski prizvyšča" by J.K.Reďko and dictionary "Prizvyšča zakarpatskych ukrajinciv: Istoryko-etymologičnyj slovnyk" by P.Čučka. The purpose of this study is to illustrate the origin and history of the Ukrainian family names development and to examine family names word-making together with their grammatical and pragmatical special features. My analysis of individual word-making styles has proved that suffixation is the most common way of word-making in the Ukraine. Furthermore, this study deals with the variety and semantics of the Ukrainian family names. Special attention is being paid to family names of the Carpathian area, which are in typological terms (word roots of non-Slavonic origin) substantially different from family names coming from central and eastern areas. I have carried out detailed analysis of the most productive patronymic suffix in Carpathian Ukraine: "- / - / -" ("-ák" in Czech). I have...
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Streptomycetes surface growth and differentiation on inert microbeads- morphology and proteome study
Tesařová, Eva ; Weiser, Jaroslav (advisor) ; Lichá, Irena (referee)
Streptomyces, filamentous Gram-positive bacteria are producers of more than 70% of antibiotics used in human therapy and agriculture. They are remarkable because of their complex life cycle (morphological differentiation) which leads to a formation of dormant spores able to survive unfavorable living conditions and allowing long-term survival of the organism. Soil represents their mostly natural living environment. In laboratory conditions they are cultivated in liquid media or on agar. We have developed in our laboratory two phase cultivation system which allows quantitative and reproducible preparation of samples for proteomic, transcriptomic and metabolomic analyses of Streptomycetes differentiation. The system is composed of inert micro- beads submerged in liquid medium. We used two types of micro-beads in our studies, glass and zirconia/silica beads. We followed the surface growth and differentiation of Streptomycetes on both types of beads using optical and electron microscopy (SEM) techniques. We observed major growth and higher antibiotic production on glass beads. Another difference we observed was in size and shape of colonies. In further research, using comparative proteomics, we attempted to identify proteins which might be responsible for recognition and adhesion of Streptomycetes to...
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Diagnosing vascular prosthesis infection by means of the FDG-PET/CT
Špaček, Miroslav ; Bělohlávek, Otakar (advisor) ; Vojáček, Jan (referee) ; Gregor, Zdeněk (referee)
Introduction: Vascular prosthesis infection (VPI) is a life-threatening complication in 0.5- 5% of cases. Early and reliable diagnosis is a necessity for adequate treatment. Computed tomography (CT) is the gold standard diagnostic method used world-wide with excellent results, but in cases of advanced graft infection. Low grade infections in non-acute patients are a diagnostic challenge requiring a new method with good diagnostic accuracy. The aim of this work is to define the accuracy of 18 F-FDG PET/CT in these settings and to identify essential parameters of its evaluation. Material and Methods: Seventy-six consecutive patients with total of 96 vascular prosthetic grafts in which infection was suspected underwent PET/CT examination prospectively. PET/CT scans were analysed in terms of presence and intensity of focal and diffuse FDG uptake, presence of anastomotic pseudoaneurysm, irregular boundary of infiltration, combination of these, and uptake ratio between graft and blood background. Also, other parameters like white blood cell count, C-reactive protein, and microbiological cultures were tested. The final diagnosis (gold standard) was based on operative/histopathological finding or >6 months clinical follow up. Results: In our series the prevalence of VPI was 57.3 % (55/96 prostheses). Among various...
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Influence of various systems on housing and breeding of dairy cattle
DUBOVÁ, Dagmar
The trend in development of new technologies and machinery in the area of agriculture and animal husbandry shows an increasing number of robotized operations. Such trend mostly appears in dairy cattle breeding, which has its logical explanation due to the fact that high ratio of human labour in milk production is more than obvious. The basic prerequisite for a successful utilization of milking robotic systems is to provide dairy cattle with sufficient quantity of top quality food and water as well as to ensure their welfare. The aim of the bachelor?s work was to compare the influence of both stanchion housing using a milking pipeline method and loose housing with robotic milking system on basic categories of dairy cattle behaviour, milk efficiency and reproduction. The observation itself was executed in each housing system for the period of 24 hours, during which an interval method with a ten-minute interval mode was used. Basic life activity of dairy cattle such as: feed intake, standing, lying and movement were monitored. All data were collected and assessed under running conditions on a private dairy farm owned by Václav Dub in Boubín near Horažďovice. The task was carried out in a reconstructed housing, where both systems are in operation. During the day, the dairy cattle remained mostly in a lying position, from which 50,33 % were kept in stanchion housing and 49,54 % in loose housing. When being housed in a standing position, 23,78 % of the dairy cattle paid attention to feeding and 27,05 % of dairy cattle were robotically milked. Standing position represented 25,89 % in stanchion housing and 19,31 % in loose housing. The movement was only monitored with dairy cattle housed in free housing representing 4,1 %. After the milk efficiency was assessed, there were some statistically significant differences (P?0,001) found in milked milk as well as in a kilogram of protein. With respect to reproduction, the length of the insemination interval was recorded showing 129 days when being stanchion housed and 95 days when loose housed. The length of the service period of dairy cattle in stanchion housing was 144 days while dairy cattle milked robotically showed the length of 109 days. All in all, it was proved that a free movement has a positive influence on the milk efficiency, fertility as well as the welfare of dairy cattle.
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