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The role of microRNAs in regulation of mammalian oocyte and embryo development
Marcollová, Kateřina ; Procházka, Radek (advisor) ; Svoboda, Petr (referee)
Cumulus-oocyte complex (COCs) is crucial for mammalian reproduction. Cumulus cells not only nurture the oocyte; they also represent important communicational nodes for mediating information towards and from the oocyte. The non-coding miRNAs can modulate posttranscriptional events they might serve as a useful biomarker for evaluating cell conditions. Based on the COCs staining with vital Lissamine Green B stain we divided cumulus cells into high- and low-quality ones. Furthermore, we implemented division based on the maturation stage, the GV and MII. Sequencing data analysis showed that DE miRNAs from qualitatively different stages donot significantly vary. Nonetheless, significantly DE miRNAs were detected between two developmentally different stages. We identified e.g. ssc-miR-183, ssc-miR-182, and ssc-miR-21-5p to be highly downregulated when comparing GV to MII stage. Among the highly expressed miRNAs from all samples were members of let-7 family (let-7c, let-7a, let- 7f-5p), ssc-miR-16, ssc-miR-21-5p, and ssc-miR-125a. Targeted genes by the DE miRNAs were involved in ErbB, TGF-β, MAPK, FoxO, gap junction and cGMP signalling pathways. We conclude that single miRNAs in cumulus cells probably cannot be used as a reliable oocyte quality marker. On the other hand,changes in the miRNA expression in...
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Effect of on in vitro maturation of porcine oocytes
Laštůvková, Viktorie ; Petr, Jaroslav (advisor) ; Benc, Michal (referee)
Lysmeral (Lilial, Butylphenyl Methylpropional) is a synthetically produced aldehyde. It is used for its typical lily of the valley-like scent as a perfume ingredient in cosmetic products. It is typically found, for example, in cleaning products. It is on the list of the 26 most well-known synthetic allergens and has been shown to be present in human urine and subsequently in wastewater. According to some opinions, lysmeral should be classified as an endocrine disruptor, which are substances that can interfere with the body's endocrine system. As these substances often affect mammalian reproduction, it is in society's interest to address them, given that they are commonly found in the environment. Lysmeral was not properly tested before being placed on the market and all its effects on organisms are still unknown. The hypothesis of this thesis is that exposure to lysmeral causes undesirable changes in the meiotic maturation of porcine oocytes in vitro and the aim is to investigate the effect of lysmeral, at concentrations corresponding to normal human exposures, on specific markers of meiotic maturation of porcine oocytes. The results confirm the negative effect of lysmeral on the smooth progression of meiotic maturation and the achievement of the final stage of porcine oocyte maturation in vitro....
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: Investigation of lymphocyte populations in the porcine small intestine
Kárová, Kristýna ; Šinkora, Marek (advisor) ; Javorková, Eliška (referee)
8 ABSTRACT Historically pig is allocated to a group of animals which use certain parts of their small intestine to acquire a fully developed primary B cell reperoire. Development of such primary repertoire is independent on the antigen presence and resembles the primary lymphopoietic activity of avian bursa of Fabricius. However, some findings concernig the pig's alignment in the above mentioned group suggest otherwise. This graduation thesis is focused on the investigtion of lymphocyte populations and subpopulations in the small intestine of germ-free and conventional piglets. The aim is to determine whether the percentage amounts of lymphocyte populations is dependent on the intestinal colonization. Using Flow Cytometry the significant differences between individual samples were assesed allowing us to conclude which parts of the small intestine could possibly be used for the development of B cell repertoire. Moreover, the status of isotype switching of B lymphocytes isolated from different intestinal parts was determined by the means of PCR analysis. Our data suggest that the small intestine colonization has a crucial role in development of all the main lymphocyte populations as well as some of their subpopulations. The greatest influence of colonization was observed concerning B lymphocytes and their...
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Characterization of the porcine induced pluripotent stem cells
Svobodová, Eliška ; Vodička, Petr (advisor) ; Tlapáková, Tereza (referee)
Induced pluripotent stem cells (iPSCs) are the cells established by introducing several transcription factors into the somatic cells and culturing them in embryonal stem cell (ESCs) culture conditions. Factors used for the establishment of the first iPSCs are OCT3/4, SOX2, KLF4 and c-MYC. IPSCs created by these means resemble closely to the ESCs. IPS technology may be used to derive iPS cells of individual pacients and apply these cells for their treatment in the cases where the use of ESCs represents an ethical and imunological problem. Therefore, it is important to establish an appropriate animal model for the longtime safety testing of iPSCs before acceding to their medicinal aplication.
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Methods and consequences of some neurosurgical interventions in animal model and in human clinic. Pinealectomy and intracerebroventricular application.
Řezáčová, Lenka ; Tichý, Michal (advisor) ; Szárszoi, Ondrej (referee) ; Přibáň, Vladimír (referee)
Clinical surgical techniques of pinealectomy and intracerebroventricular (i.c.v) cannulation are still used in the clinic in indicated cases. In research, there is an effort to replace these classical surgical techniques by other means. However, these various new models often do not reflect the complexity of the functions taking place in the living organism as a whole. The thesis shows that these surgical techniques should be also a part of biomedical research in the future, as they still bring new important information. In the experiment, the pinealectomy can serve as a model of resection or depletion (removal of natural hormone secretion), while i.c.v. application of the active substance to the ventricles on the contrary as a model of addition or substitution. The practical part of the thesis is divided into 4 areas: A) pinealectomy - an animal models, B) pinealectomy - in the experiment, C) pinealectomy - in the clinical practice and D) intracerebroventricular application - in the experiment. The work describes in detail the surgical techniques and discuss possible consequences of pinealectomy in 6 animal species (and their comparison) and in humans. It also includes an experiment with pinealectomy in a rat and a clinical study in patients with pineal cyst and subsequent pinealectomy. The work...
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Effect of sperm ubiquitination in early embryonic development of porcine embryos
Petelák, Aleš
The PhD thesis is focused on the effect of porcine sperm cell extracellular ubiquitination on early embryonic development up to the blastocyst stage after ICSI. In addition, it also presents a potential improvement of the technique of in vitro fertilization using oocyte incubation with ion channels regulators. To address these aims, we established an entirely novel methodology for sperm cell sorting using flow cytometry and subsequent cryopreservation. We determined the conditions for successful sperm cell sorting based on extracellular ubiquitination rate providing highly specific selection as well as sufficient numbers of viable sperms for fertilization using the ICSI method. Concerning the following cryopreservation, established methods were optimized to enable freezing of a minimal sperm cell suspension volume with low cell numbers. The performed experiments showed a direct relationship between the rate of extracellular ubiquitination and the capability of sperms to give rise to a properly developing embryo. Highly ubiquitinated sperm cells were less successful regarding the embryonic development to the blastocyst stage if compared with the lowly ubiquitinated group (6,2 % vs. 16,7 %, P<0,001). Interestingly, the rate of extracellular ubiquitination showed no effect on the pronuclear formation...
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Effect of sperm ubiquitination in early embryonic development of porcine embryos
Petelák, Aleš ; Krylov, Vladimír (advisor) ; Šušor, Andrej (referee) ; Procházka, Radek (referee)
The PhD thesis is focused on the effect of porcine sperm cell extracellular ubiquitination on early embryonic development up to the blastocyst stage after ICSI. In addition, it also presents a potential improvement of the technique of in vitro fertilization using oocyte incubation with ion channels regulators. To address these aims, we established an entirely novel methodology for sperm cell sorting using flow cytometry and subsequent cryopreservation. We determined the conditions for successful sperm cell sorting based on extracellular ubiquitination rate providing highly specific selection as well as sufficient numbers of viable sperms for fertilization using the ICSI method. Concerning the following cryopreservation, established methods were optimized to enable freezing of a minimal sperm cell suspension volume with low cell numbers. The performed experiments showed a direct relationship between the rate of extracellular ubiquitination and the capability of sperms to give rise to a properly developing embryo. Highly ubiquitinated sperm cells were less successful regarding the embryonic development to the blastocyst stage if compared with the lowly ubiquitinated group (6,2 % vs. 16,7 %, P<0,001). Interestingly, the rate of extracellular ubiquitination showed no effect on the pronuclear formation...
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Effect of sperm ubiquitination in early embryonic development of porcine embryos
Petelák, Aleš ; Krylov, Vladimír (advisor) ; Šušor, Andrej (referee) ; Procházka, Radek (referee)
The PhD thesis is focused on the effect of porcine sperm cell extracellular ubiquitination on early embryonic development up to the blastocyst stage after ICSI. In addition, it also presents a potential improvement of the technique of in vitro fertilization using oocyte incubation with ion channels regulators. To address these aims, we established an entirely novel methodology for sperm cell sorting using flow cytometry and subsequent cryopreservation. We determined the conditions for successful sperm cell sorting based on extracellular ubiquitination rate providing highly specific selection as well as sufficient numbers of viable sperms for fertilization using the ICSI method. Concerning the following cryopreservation, established methods were optimized to enable freezing of a minimal sperm cell suspension volume with low cell numbers. The performed experiments showed a direct relationship between the rate of extracellular ubiquitination and the capability of sperms to give rise to a properly developing embryo. Highly ubiquitinated sperm cells were less successful regarding the embryonic development to the blastocyst stage if compared with the lowly ubiquitinated group (6,2 % vs. 16,7 %, P<0,001). Interestingly, the rate of extracellular ubiquitination showed no effect on the pronuclear formation...
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Effect of selected factors affecting the percentage of lean muscle in pig carcass
KOLÁŘOVÁ, Eva
The aim of the thesis was to obtain information on the effect of hybrid combination, gender and weight of cold JUT on carcass ratios in final pig hybrids. 1742 pig carcasses of four hybrid pig combinations were included in the carcass monitoring: PIC, Topigs, Danbred and France Hybrides. The average carcass weight was 89 kg with an average muscle mass of 59.5%, represented mostly by quality classes S and E. The higher fat content of pigs versus gilts was shown. With increasing slaughter weight, the proportion of muscle decreased and the proportion of fat increased.
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Chov prasat plemene bílé ušlechtilé v České republice
ROHÁČEK, Daniel
The goal of this bachelor thesis is to analyze the results that have been provided by the Czech Association of Pig Breeders regarding productivity of Czech Large White breed. For the ob-served period of 1999-2017, the intention was to compile the tables that would summarize the number of litters, number of born piglets in one litter (all vs. live born), number of piglets sur-viving 21 days, weight of piglets after 21 days, and average time between 2 litters. The source materials and the used literature clearly indicate that the swine production in the Czech Republic is shrinking. Swine breeders are closing their production due to instable price of pork meat. The number of sows decreased by 177 755 between 1999 and 2017. As a con-sequence, the number of litters declined as well. On the other hand, this impact is compensa-ted by permanently shortening time distance between two consecutive litters, which currently stands at 156 days. This logically results to increasing number of litters per sow, now amoun-ting to 2.4 per year. Effective breeding also leads to increasing number of piglets in one litter. In 2017, one sow brought in average 14.8 piglets in a litter, of which 13.4 live born. Also the number of successfully bred piglets is growing thanks to improving technologies and better trained pers onnel, having reached 11.7 piglets per litter in 2017. Sows of Czech Large White breed deliver increasing lactescence, as the weight of a litter after 21 days grew in the obser-ved period by 13.4 kg to 69.1 kg. In the next part of the thesis, analysis is made for field tests and results of the unified test, both supplemented by tables and charts. For the observed period, the goal also was to evaluate the average daily weight gain, the percentage share of muscle mass, and the average thickness of saddle fat in mm of both sows and boars. It was found out that the Czech Large White breed had an excellent meat utilization. The share of lean muscle is increasing continuously - in 2017 it reached 63.6 % for boars and 63 % for sows. The vast majority of supplied pigs was classified into the highest-quality categories. The final part of the thesis contains the list of farms that were certified for breeding the Czech Large White breed in 1999 - 2017. The number of such farms was growing until it rea-ched 24 in 2001 and 2002. However, after subsequent decline there was only 6 such farms in 2017.
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