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Dynamics of mouse sperm capacitation and acrosome reaction
Dvořáková-Hortová, Kateřina ; Frolíková, Michaela ; Děd, Lukáš ; Šebková, Nataša
Capacitation followed by the acrosome reaction (AR), is a very complex event of molecular changes, including acrosome matrix rearrangement and actin polymerization, which mammalian sperm must undergo in the female reproductive tract in order to obtain the ability to penetrate and fertilize the egg. CD46 and β1-integrin belong to specific proteins, which are predicted to interact during molecular reorganization of capacitating sperm. The IZUMO1 as the primary fusion protein of the mammalian sperm is also involved in this dynamic network. We investigated the relationship between the Izumo, CD46 and β1 integrin relocation in the sperm head during the capacitation and AR in vitro. We have already successfully monitored by immunofluorescent labelling the dynamics of proteins CD46 and β1-integrin. The changes in the localization of these proteins associated with the AR and their mutual co-localization was observed. The original β1-integrin location in the freshly released epididymal sperm is in the acrosome and it relocates during the AR further through the sperm head compartments into the equatorial segment and over the whole sperm head. Its density over the equatorial segment is decreasing with the extended time of the AR. Also its presence in the perforatorium of the mouse sperm head is very prominent. The pattern for protein CD46 is extremely similar if not identical in both aspects such as compartment localization and time progress during capacitation and AR in vitro. The molecular interaction of CD46 and β1-integrin was investigated using the Proximity Ligation Assay and Super resolution microscopy STED. The data were statistically analysed. The newly obtained results from CD46 and β1-integrin relocation are in correlation with IZUMO1 dynamics and giving a substantial knowledge on the studied protein network rearrangement during capacitation and AR in mouse spermatozoa.
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Work in social integration process of adults with intellectual disability.
SVITÁKOVÁ, Iveta
In my bachelor thesis, I dealt with the use of work activities in the process of social integration of adults with mental disabilities, the aim was to create, implement and evaluate regular work activities intended to a specific group of adults with mental disability and focused primarily on the development of social integration. The thesis is structured into six chapters. The first of them is focused on the definition of mental disability, characteristic of adulthood as an important developmental period in human life, and on adulthood of individuals with mental disabilities with a goal to find and identify the specifics that were needed to be taken into account when working with this target group of people. The following section is devoted to the social integration, its determinants and the environment in which it operates. I continued with the outputs in the third chapter, where I focused on key competencies, which them are the necessary basis for social integration, especially social and personal skills and communication skills, necessary basis. Targeted professional development can be realized through educational activities, but has to respect the specificities of the participants and voluntary participation. Both of these assumptions can be fulfilled within the leisure education using adequate pedagogical methods. For this reason, I decided to focus my attention on these areas at the end of the chapter. The fourth chapter is devoted to work and work activities, namely the definition of concepts, their characteristics and importance and role of work activities in human life, or a person with mental disability. Based on all acquired theoretical knowledge and using previous experience with the target group in the fifth chapter, I designed a specific training program based on the work and activities focused on the development of selected key competences. The educational program volunteered four participants, but I have decided to follow the development of competencies of three participants because one participant attended the program very irregularly. The program has been implemented three times a week, two and a half hours for seven months (from October 2015 to May 2016) in the premises of the training centre Mo-zai-ka. At the beginning of the realisation of the created educational program, the participants agreed that we will adhere and develop good habits necessary for good relations within the group and the results of the work. On a basis of the individual characteristics of the participants, I prepared the work, which should also contribute to the development of social and personal and interpersonal skills. I have continuously monitored and recorded them. During the implementation of the program, I put emphasis on repetition, patient negotiations with participants and creating a friendly atmosphere. After completing the training program, I evaluated the development of competencies in three selected participants. On that basis, I concluded that the biggest change was in the communicative competence. I also experienced a change in social and personal competencies, however, results varied widely among individual participants. Yet, I believe that the goal of the thesis was achieved. These results show that education in leisure time, based on work activities may lead to the development of social and personal and interpersonal skills that are for adults with mental disabilities necessary in the process of social integration. This thesis can be an interesting source of inspiration for the realisation of similar programs aimed at social integration of persons with mental disabilities.
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Narcolepsy and other seizure disorders
TONDLOVÁ, Andrea
The thesis deals with the issue of seizure disorders from the perspective of special neurology. In terms of content, it is focuses on two major areas. The first part is theoretical in nature and has descriptions of the individual disorders, their clinical manifestations, diagnoses and current treatments. It is focused on the following seizure disorder group with regard to one of the objectives of the work: epilepsy, tetany, Meniere's syndrome, migraine and more specifically narcolepsy. These findings are supplemented with information on current knowledge regarding the treatment and diagnoses of these diseases, which make the lives of thus disables people very difficult and prevent them from leading their lives the way they could if they were in full health. The aim of the work is not just a theoretical description of the currently available knowledge of these diseases from literature, but also a practical confrontation thereof from experiences, which in this case result from the true stories of people suffering from seizure disorders. A specific seizure disorder has been selected for this second part of my thesis, which needs to be talked about more in order to become better known. It is a disorder that is interchangeable with regard to its symptoms, which are similar to other disorders, and it is difficult to diagnose or is only diagnosed after an inconveniently long time - i.e. Narcolepsy . The thesis therefore deals with the issue of narcolepsy in most detail, with regard to the practical focus in the next section, where in addition to aforementioned basic knowledge, the author describes the activities and the importance of sleep laboratories and centres as institutions involved in treatment. In order to penetrate into the issue of this disorder even more deeply, she adds the aforementioned anonymous stories of people affected in this way. This part is also complemented with illustrative images of the sleep laboratory and the stories of these people as a better visual complement of the thesis for the reader. A descriptive analysis (comparison) of each symptom was chosen as a comparative method of the aforementioned thesis objective (to compare the theoretically given symptoms of narcolepsy with the symptoms of individuals suffering from this disease), in which the goal was to choose the common symptoms these people which make their lives difficult and compare these data with information (symptoms of narcolepsy) found in contemporary literature. In conclusion, we visit the most important information from scientific literature, the views of the professional community on the current research and studies on narcolepsy, i.e. the specifically examined causal links of applying the Pandemrix vaccine (against swine flu) and narcolepsy and a summary of the results of other studies investigating the etiology of this disease. The aim of this section is to present the reader with, and identify, the current developments in narcolepsy research. An evaluation of the historical development of interest in sleep disorders and an evaluation of the conclusions drawn by comparing symptoms specified in professional literature on narcolepsy with actual symptoms of people suffering from this disease are also included. The aim is to provide the reader with a bio-psycho-social view of a person with this disease, which is not always emphatically discussed in every piece of professional literature. The result of the entire thesis is a summary of the current state of knowledge of these diseases and a reflection on the research of the current possible causes of narcolepsy. It continues to compare the theoretical background of the narcolepsy disorder with the experience of living with this disease, including stories of selectively chosen individuals suffering from narcolepsy.
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Methodist School-Based Prevention and his Role in Practical Elementary School and Special Elementary School
HAVLÍČKOVÁ, Markéta
The diploma thesis focuses at a theme of Methodology of school prevention and its role at the specialised primary school. The theoretical part is split into four main chapters which refers to the education system of children with need of special educative care in the Czech Republic. It defines socially pathological aspects including risk behaviour with its prevention. The last chapter describes primary prevention at schools using a minimal preventive program and work of a methodologist of school prevention. The practical part contains a quality-type research based on semistructural interviews with methodologists of school prevention at chosen specialised primary schools. The first task of my diploma thesis is to find out what advise parents need and ask for the most. The other task is to find out what aspects a methodologist of school prevention has to take into consideration during issuing a minimal preventive program. The questions have been asked in the way to find out how cooperation at the level of prevention between parents and specialised primary school works, and who else among school staff except of a methodologist participates in the issuing a minimal prevention program. I have contacted sixteen specialised primary schools in the South Bohemia region for the interview. Ten have replied, but only six of them were willing to cooperate further. Two of the schools are located in the regional city, the other four are from smaller cities of similar sizes. Interviews were carried out with methodologists of school prevention and questions were focused at the cooperation at the level of prevention between parents and specialised primary school, and possible influence of location of the school which could be reflected in the minimal prevention program.
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The effect of pyrethroid based pesticides on fish
RICHTEROVÁ, Zuzana
Pyrethroids are ones of the most used pesticides worldwides. The widespread use and high stability of pyrethroids lead to the assumption of that their occurrence in the environment could be quite frequent. They can reach water ecosystem as pollutants. Residues of pyrethroids are not only detected in the water column, but also in sediments and in fish tissues. The first study was devoted to the product Nexide containing 60 g.l-1 of active substance gamma-cyhalothrin. Tested Nexide concentrations were 5, 25, 50, 100, and 250 µg.l-1. Early life stage test was used.Common carp (Cyprinus carpio) was tested. There were significant mortalities in all concentrations except the lowest concentration during the trial. The lowest concentration tested 5 µg.l-1 only caused a slightly increased mortality. This lowest concentration influenced the growth in length and weight negatively, decelerated ontogenetic development, and made the body surface of the individuals darker. Histopathology of individuals from this concentration revealed dystrophy in liver. Examination of kidney, intestine and gills did not show significant histopathological differences compared with control. The evaluation of selected parameters of oxidative stress demonstrated a significantly higher activity of detoxification enzyme glutathione-S.transferase (GST) and a significantly lower activity of defensive enzyme glutathione peroxidase (GPx) compared with the control group. The other examined parameters of oxidative stress such as catalase (CAT), glutation reductase (GR), and lipid peroxidation determined by using the thiobarbituric acid-reactive substances (TBARs) were comparable to the control group. Changes in oxidative stress parameters suggest that exposure of the organism to the product Nexide in the given concentration leads to dysbalance of defensive enzymes. The second study was devoted to the product Cyperkill 25 EC containing 250 g.l-1. Tested Cyperkill 25 EC concentrations were 7.2, 36, 72, 144, and 360 µg.l-1. The procedure of the trial was the same as the preceded one. There were 100% mortalities in all concentrations except the lowest concentration during the trial. The lowest tested concentration 7.2 µg.l-1 allowed 90% of individuals to stay alive till the end of experiment. The lowest concentration influenced the growth in length and weight negatively and decelerated ontogenetic development compared with the control. Any individual exposed to this concentration did not reach juvenile stage until the end of the trial. Dark pigmentation was visible in 68% of these exposed individuals on the last day. Similar darkening was visible in individuals from higher concentrations shortly before death too. Histological examination did not revealed significant changes in intestine, liver, kidney, and gills compared with the control group. Evaluation of selected parameters of oxidative stress demonstrated significantly lower activities of GST, GR, and GPx. Activities of CAT and TBARS were comparable with the control group. Changes in oxidative stress parameters suggest that exposure of the organism to the product Cyperkill 25 EC in the given concentration could induce oxidative stress and interfere with the activities of antioxidant enzymes. The presented thesis summarises actual data about pyrethroids and their influence on fish. The demonstrated effects confirm high susceptibility of early developmental stages of fish to tested pesticides. When interpreting the results, we have to take into account the fact that studies showed this risk even on single pyrethroid substances. But water organisms are exposed to many other more or less toxic products and substances in a real environment. These xenobiotics could react with each other and their mixture could even potentiate negative effects. The performed studies also clearly show the significant differences in the sensitivity of embryonic and embryolarval tests.
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Iniciace pohybu byčíku, signalizace a regulace pohyblivosti spermií ryb: fyzikální a biochemické řízení
PROKOPCHUK, Galina
The current study attempted to shed light on the regulatory processes and response arrangements of fish spermatozoa during the course of maturation and motility initiation. The first intent of this study was to improve the understanding of the mechanism underlying the acquisition of potential for sperm motility in sturgeon. Up to present work, the physiological process underlying sperm maturation in this species has not been described at all. Our results showed that sperm maturation in sturgeon occurs outside the testes because of dilution of sperm by urine and involves the participation of high molecular weight substances as well as calcium ions present in seminal fluid and/or urine. The second aim of the present study was to investigate the coping mechanisms in fish spermatozoa with osmotic and ionic activating mode, as well as in spermatozoa of euryhaline fishes, to various environmental conditions. We showed that alteration of environmental osmolality might affect the fish sperm in different ways, depending on fish species and modes of spermatozoa motility activation either osmotic or ionic mode. In response to osmotic stress caused by hypotonicity, carp spermatozoa regulated the flow of water across their cell membrane and increased their cytoplasmic volume during their short motility period. In contrast, no indications of sperm volume changes were observed neither in sterlet nor in brook trout spermatozoa, both of which having an ionic mode of motility activation. We also examined the mechanism by which sperm motility triggering in euryhaline fishes can adapt to a broad range of environmental salinity. Our results demonstrated that spermatozoa of euryhaline tilapia, Sarotherodon melanotheron heudelotii, reared in fresh-, sea- or hypersaline water can be activated in hypotonic, isotonic or hypertonic conditions of swimming milieu, provided Ca2+ ions are present at various levels. It was established that the higher the fish rearing salinity or the more hypertonic ambient media at spermatozoa activation, the higher extracellular concentration of Ca2+ ions is required. The results obtained in the present study allow suggesting that osmolality is not the main factor inhibiting sperm motility inside the testis in the S. melanotheron heudelotii. A third aim of this study was investigation of the regulation of motility initiation process and description of flagellar beating initiation in chondrostean spermatozoa. We detected that K+ inhibition of sperm motility in sturgeon can be by-passed due to the pre-exposure of sperm cells to a high osmolality shock prior to its transfer to K+-rich swimming media. Thus, we hypothesized that sturgeon spermatozoa may be activated by use of an unexpected signaling pathway, independent from regular ionic stimulation. The successive activation steps in sturgeon spermatozoa were investigated by high-speed video microscopy, using specific experimental situation, where sperm motility initiation was delayed in time up to several seconds. At motility initiation, the first couple of bends formed at the basal region begins to propagate towards the flagellar tip, but gradually fades when reaching the mid-flagellum. This behavior repeats several times until a stage where the amplitudes of bends gradually reach similar value, what eventually leads to sperm progressive displacement. The total period needed for the flagellum to switch from immobility with rigid shape to full activity with regular propagating bends ranges from 0.4 to 1.2 seconds. In conclusion, the results of the current study bring valuable pieces of information into the general understanding of the processes of maturation of fish spermatozoa, their adaptability to different physical and biochemical circumstances, the extra- and intra-cellular signaling as well as the regulatory mechanisms of motility activation in fish spermatozoa.
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Some Robust Distances for Multivariate Data
Kalina, Jan ; Peštová, Barbora
Numerous methods of multivariate statistics and data mining suffer from the presence of outlying measurements in the data. This paper presents new distance measures suitable for continuous data. First, we consider a Mahalanobis distance suitable for high-dimensional data with the number of variables (largely) exceeding the number of observations. We propose its doubly regularized version, which combines a regularization of the covariance matrix with replacing the means of multivariate data by their regularized counterparts. We formulate explicit expressions for some versions of the regularization of the means, which can be interpreted as a denoising (i.e. robust version) of standard means. Further, we propose a robust cosine similarity measure, which is based on implicit weighting of individual observations. We derive properties of the newly proposed robust cosine similarity, which includes a proof of the high robustness in terms of the breakdown point.
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The Human Resources and Employment Operational Programme - specific example
Novotná, Jitka ; Hrdlička, Pavel (advisor)
The theoretical part is focused on characteristics of the labor market , causes and consequences of unemployment risk groups of people over 50 years , the European Social Fund and its impact on the labor market and characterized Operational Programme Human Resources and Employment 2007-2013.
At the conclusion of the theoretical part are specified individual regional project Key competences in Hradec Kralove region.
In the practical part of the thesis evaluates the individual regional project Key competences in Hradec Kralove region and described its own survey of long-term success of the project operators in the labor market and measures designed to increase the success of similar projects .
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Genetic eye diseases in dogs
Šmejkalová, Barbora ; Hofmanová, Barbora (advisor)
Eye diseases in dogs are often very similar to humans eye disease. Eye diseases are becoming more common in pure breed of dogs, therefore it is important to take them into consideration, particularly when we are crossing the parent generation.
In the first part is explained in detail and illustrated anatomy of the eye, including images and photographs. In the next part, there are described various eye diseases, namely cataract, glaucoma, hereditary diseases of the retina, progressive retinal atrophy (PRA) and Collie Eye Anomaly (CEA). For each disease is described, how it is possible to diagnose the disease, how it progress, what is its connection with genetic and possible treatment alternatives. In closing there are tables and graphs showing the frequency of occurrence of a specific disease either from an overall perspective or from the perspective of individual breeds.
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Effect of dairy cows housing technology on mastitis occurrence
Brzáková, Lenka ; Stádník, Luděk (advisor) ; Jan, Jan (referee)
The objective of my bachelor thesis was evaluation of quality of housing, milking and making analysis of mastitis on chosen farm.
First part of the thesis contains of literature summary about housing technologies, mastitis issues, their detection, originators, therapy and factors affects against their incidence. Thereafter is this part focused on tipes of milking parlours, corect milking procedures and mechanized milking.
Materials and methodes of the thesis contains the characteristics of chosen farm where the quality and technology of housing, milking and nutrition of milk cows and other factors affecting presence of mastitis such as taking care of cloven hooves was watched in 2014. The incidence, detection and curation of mastitis was watched afterwards. Montamilk s.r.o. farm cradles 1,200 pieces of cattle and milk cows are about 419 out of it.
In the part of thesis named results was watched and evaluated inspection of efficiency in inspecting year 2013 - 2014, which was stated on first lactated cows and on cows which are on second and further lactation. Milk yield was far better with cows on second or futher lactation according to data gathered during the study and expressed by diagram. Diagram of milk components (fat and proteins), which was divided by lactation, was expressed in this part too. Other examinated parameters was occurrence of mastitis - which milk cows, in which season is the occurrence more often, treatment and cost of treatment for one cow. At the end the number of somatic cells for 2014 was represented in diagram for particular lactations.
In part of thesis called discusion the comparison was drowed between results from chosen farm and national average of efficiency checks. From results is obvious that milk cow in Mnotamilk has been better in first lactated cows by 427 kg and in second and more lactated cows by 602 kg than is the avarage in Czech republic.
Far more the milk components in Montamilk was compared with rest of Czech republic and the results was quite similar. The volume of fat of Montamilk cows was lower in every lactation in comparison with milk cows from Czech. Value of fat in Czech republic avarage first lactated cows is 3,87 % in Montamilk 3,85 % and the value of fat on second and more lactation cows is 3,86 % in Czech average and 3,81 % in Montamilk.
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