Malý, L.: Study of biological activity of alkaloids isolated from Fumaria officinalis L. (Fumariaceae) II. Diploma Thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2014, 49 pp. Obtained diethylether extract of Fumaria officinalis L. was separated to fractions in column chromatography with petrol, chloroform and ethanol. Preparative TLC and crystalisation led to isolation of five alkaloids from fraction. Alkaloids were identified by GC-MS and NMR specters, optical rotation and melting point as protopine, cryptopine, (-)-fumaricine, (+)-fumariline and (+)-parfumidine. Isolated alkaloids were tested for their inhibition activity towards acetyl- and butyrylcholinesterase and towards prolyloligopeptidase. Activities were compared with standards. Natural inhibitor galanthamine showed IC50 AChE 1.710 ± 0.065 µM, IC50 BuChE 42.30 ± 1.30 µM. Best inhibition activity showed protopine (IC50 AChE 345.4 ± 24 µM, IC50 BuChE 239.6 ± 22.3 µM) and cryptopine (IC50 AChE 477.71 ± 47.33 µM, IC50 BuChE 270.82 ± 39.12 µM). The highest prolyloligopeptidase inhibition activity showed (+)-parfumidine with IC50 POP 99.2 µM, which was more active than used natural inhibitor baicaline (IC50 POP 605.9 ± 0.021 µM). Synthetic POP...

Kostelník, J.: Study of biological activity of alkaloids isolated from Fumaria officinalis L. (Fumariaceae) I. Diploma thesis, Charles University in Prague,Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2014, 63 p. The aim of this study was to isolate alkaloids from joined fraction no. 55-67 (A2) obtained from the total alkaloid fraction of extract of Fumaria officinalis L. (Fumariaceae) plant. Using chromatography methods three alkaloids were isolated and then identified by structural analysis (GC-MS, NMR). Three alkaloids were isolated by using common chromagografic methods and then identified by structural analyses optical rotation and melting point as (-)-O- methylfumarophycine, (-)-sinactine a (-)-stylopine. Inhibitory activity of isolated alkaloids was assessed against human erythrocyte acetylcholinesterase, human butyrylcholineesterase and prolyl oligopeptidase. The results were expressed as IC50 values ((-)-stylopine: IC50 AChE and IC50 BuChE > 1000 μM, IC50 POP > 1000 mM; (-)-O-methylfumarophycine: IC50 AChE = 963.10 ± 135.98 µM, IC50 BuChE = 1771.0 ± 380.94 µM, IC50 POP - unmeasured; (-)-sinactine IC50 AChE = 632.0 ± 68.12 µM, IC50 BuChE = 8154.3 ± 981.42 µM, IC50 POP = IC50 POP = 52.9 ± 1.8 µM). None of alkaloids isolated showed...

The inter-population geographic song variability can be found in many bird species. One of the most interesting types are dialects, characterized by sharp boundaries between populations of individuals sharing the same particular song type. Despite many years of investigation there is no satisfactory answer to the question how these borders persist over the years at nearly same sites. Distinctions between dialects allow recognition of males from different populations and could influence the territorial behaviour. Various habitats, however, degrade the sound differently; it is therefore possible that specific habitats are preferred by individuals with specific dialects. Both social interactions and acoustic properties of the locality could also be only secondary factors and the dialect distribution could be the result of the habitat distribution either in present or in recent past. In this diploma thesis, several hypotheses were tested on the example of Yellowhammer (Emberiza citrinella) - a songbird abundant in open land with simple song and easily recognizable dialects. The thorough data concerning the habitats and dialect distribution were obtained at the dialect borders at two localities in Czech Republic. The reactions of territorial males to both foreign and their own dialects were also...

Michal, Vojtěch: STUDY OF BIOLOGICAL ACTIVITY OF ISOLATED ALKALOIDS FROM ARGEMONE GRANDIFLORA (PAPAVERACEAE) II. Diploma thesis 2015. Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology. Supervisor: PharmDr. Jakub Chlebek, PhD. Key words: Argemone grandiflora Sweet, Papaveraceae, alkaloids, isolation, acetylcholinesterase, butyrylcholinesterase, prolyloligopeptidase, Alzheimerʼs disease, in vitro assay. Diethylether alkaloid extract obtained from stem and roots of Argemone grandiflora Sweet was chromatografically analyzed. Using common chromatografic methods, three alkaloids were isolated in clean form. These substances were identified as allocryptopine, (-)-munitagine and (-)-norargemonine by structural analysis (MS, NMR). These obtained alkaloids were tested for their inhibitory activity against human erythrocyte acetylcholinesterase (AChE) and human plasma butyrylcholinestrase (BuChE) by Ellman's method. The results were represented as IC50 values (allocryptopine: IC50 AChE = 250,0 ± 2,52 μM, IC50 BuChE = 530 ± 28,2 μM; (-)-munitagine: IC50 AChE = 62,29 ± 5,81 μM, IC50 BuChE = 837,4 ± 23,03 μM; (-)-norargemonine: IC50 AChE = 205,17 ± 11,6 μM, IC50 BuChE = 4158,20 ± 495,78 μM). Inhibition against prolyloligopeptidase was tested for...

The objective of this thesis is to design a log house construction project meeting the needs of a four/five person family. The initial chapters describe a variety of up to date wooden building construction systems. Analysing their characteristics, a construction project for the wooden building is designed including the building envelope in three different modifications: frame wooden building with diffusion closed structure, frame wooden building with diffusion-opened structure, massive wooden building. For each structure of building envelope is then performed thermal technical assessment, all of which variants are designed so that the heat transfer coefficient structure was the same for each part of the structure of individual systems. The second part of the thesis deals with the investigation of the construction project itself making use of one of the above mentioned construction systems. The last part is focused on setting a price for each structure of building envelope separately.

The Master's Thesis deals with the issue of solitude of seniors. It examines the solitude from theological, psychological and sociological point of view and seeks inspiration in the Social Doctrine of the Catholic Church. It tries to understand the position of Benedict XVI. who says that the most painful kind of poverty is solitude. The acquired knowledge is then viewed through the optic of social work and possible solutions of the problem of solitude of seniors are suggested.

This thesis is focused on the role of a pediatric nurse in the care of overweight and obese adolescents. A pediatric nurse plays a key role in promoting the health of children and adolescents in primary care. A nurse is in many cases the first person the family or a child confide their worries and health concerns in. A pediatric nurse is a person whose professional focus is determined by the needs of children; he or she is an expert who should seize every opportunity for consultation on healthy lifestyles of children and parents; he or she is an essential and irreplaceable member of the team caring for a child or an adolescent (Sikorová, 2012). This thesis consists of two parts. The theoretical part is focused on obesity itself, it describes pathogenesis, complications and consequences, as well as prevention and treatment. In the next chapter we focus on the role of the nurse in the pediatrician's office. There we described the role of nurses in the examination of an obese or overweight adolescent, the awareness of a nurse and adolescent in the field of childhood obesity, the principles of communication of nurses with the obese adolescent and his or her education. At the end of the theoretical part the psychosocial impacts on obese adolescents, such as social isolation, discrimination and the risk of developing eating disorders, are described. The second half of the thesis is empirical. Research was carried out in two ways. The first part of the research was realized in the form of a quantitative survey, in which nurses in general pediatricians offices were approached using electronic questionnaires. The questions in the questionnaire were focused on the education of nurses in the field of childhood obesity, education of adolescents, cooperation with families and other professionals. The research group therefore consisted of pediatric nurses who pursue their profession in the Czech Republic. In the second part of the research the method of questioning, namely the interview technique was used. It was a semi-structured interview with obese adolescents. Each interview consisted of 24 questions and lasted an average of 20 - 25 minutes. Interviews with adolescents were focused on the knowledge of the term Body Mass Index, health and psychological consequences, their personal lives, relationships with peers and partners, the possibility of using the services of a nutritional consultant. We contacted 15 respondents aged 16 to 20 years, all were from South Bohemia. In case of minor adolescents parental agreement with the research had to be acquired. The questionnaire was accessible online and its completion was not time consuming; however out of approximately 400 distributed questionnaires only 154 returned fully completed. The return of questionnaires was approximately 40 % and reflects the ratio of completed and mailed questionnaires. At the beginning two goals were set. The purpose of the first objective was to explore the role of nurses in the approach to obese adolescents. The results showed that nurses are trying to educate themselves in the field of child obesity, they are interested in the issue of childhood obesity, what is more they are trying to educate the overweight adolescents in good nutrition. Furthermore, they also aim to cooperate with the child's family and even with other professionals, such as an endocrinologist, cardiologist, nutritionist etc. The second objective was to determine how obese adolescents perceive themselves. Opinions obviously differed, some children did not admit to being overweight/obese, while others saw themselves as the "fatsoes", "whales" ugly and useless, and that nothing suits them". Some suffer from depression, others try not trouble themselves with it. To fulfill the purpose of quantitative research 3 hypotheses were set. H1: Nurse educates adolescents about healthy eating. The hypothesis was statistically confirmed because research showed that nurses really do educate teenager.

Considering contemporary lifestyle and possibility of travelling imported infections accrued and some illnesses come back that have not occurred nearly at us more. Not only haemorrhagic fevers and other highly infectious illnesses but also hospital multiresistant bacterial strains that mean high risk for patients constitute a threat. The number of nosocomial infectious grows still. The patient is in the case of suspicion or already diagnosed infectious illness isolated and treated in the infectious department or in the tribal department in the room with increased hygienic-epidemiologic regime. The patient is saved in a single room, the possibility of visits is reduced and the staff uses personal protective equipment. The man is holistic being and it can come due to isolation to lack of satisfaction of his/her bio-psycho-social needs. The aim of this thesis is to identify unsatisfied bio-psycho-social needs of patients in increased hygienic-epidemiologic regime and to describe changes in needs of the patient depending on the time of hospitalisation. The theoretic part describes the problematics of infectious illnesses, system of the care for the patient with infectious illness and changes in human needs in the period of the illness. The empiric part of the thesis deals with unsatisfied bio-psycho-social needs of isolated individual through quantitative research. The research suite was made up of 10 patients who were hospitalised in the room with increased hygienic-epidemiologic regime. The survey was carried out in the Hospital Jihlava by semistructured interview and the results of the survey were processed by opened coding by method paper and pencil. It is emphasized that the most needs of the patient have biological, psychical, social and even spiritual aspects. The rate of satisfaction with caregiving depends on the health condition and personality of the patient. Not small influence has the length of the hospitalisation, too. It was found out during the survey that the patients have not enough information about specifics of the care in the infectious department and that is why there is often a misunderstanding of necessity of barrier measures. It is possible to avoid useless misunderstanding through timely and suitable education. It was created an educational material with the title "Guide for patients hospitalised in the infectious department of Hospital Jihlava" for this purpose. Employees of the infectious department of Hospital Jihlava were informed with results of the research during a workshop, too.

The aim of this work was the production of two Drosophila growth factor proteins, DS47 and IDGF3. The cDNAs encoding the two proteins were cloned into a baculovirus transfer vector and used for the recombination with bacmid form of baculovirus maintained in Escherichia coli. The recombinant baculoviral DNA was transfected to insect cells. Baculovirus- infected cells produced recombinant His-tagged proteins which were purified from growth media by nickel-agarose affinity chromatography. The function of recombinant proteins was verified by in-vitro growth/survival tests.

It is the purpose of this thesis to implement primordial germ cell (PGC) transplantation, one of the new biotechnological reproductive methods, and for this to explain the details that we have to know about embryo development and PGC migration in pikeperch. We provide several specific useful methods such as GFP labelling and blastodisc surgery which are required for efficiency assessment of the transplantation technique. The main results of the publications in the thesis could be informative and useful for generation of germline chimera by using pikeperch. We described pikeperch embryo development to first feeding at 15°C in detail and demonstrated effects of temperature on the rate of embryogenesis to determine temperature limits for slowing development with minimum negative effects on growth and survival rate. We also developed a technique to soften the pikeperch chorion by enzyme in order to remove it by forceps for in depth observation. Additional groups of eggs were fertilised and incubated at different temperatures to document embryo developmental stages, developmental rate, and survival. The optimum fertilisation and incubation temperature was 15°C, with the highest fertilisation, survival, and hatching rates. Embryo development was drastically slowed down at 10 °C, with 45% of fertilised embryos surviving to hatching. Development was accelerated at 20 °C, with a 56% survival rate of fertilised embryos. After the series of experiments to characterize the embryo development of pikeperch, it could be a valuable model percid for research in which flexible incubation temperatures is required. We described the important early embryonic events, namely, yolk syncytial layer (YSL) formation and midblastula transition (MBT) during the blastula stage in pikeperch embryos. The chorion was removed as we described in the first study. The YSL was formed after the breakdown of marginal cells during the 512- to 1k-cell stage. Cell division analysis by 4'-6-diaminido-2-phenylindole (DAPI) staining revealed that transition from synchronous to asynchronous division occurred after 1k-cell stage. Our results indicate that MBT starts after this stage. Next, we performed blastodisc isolation assay to find the competent stage for embryonic manipulation. Embryos were manipulated by using a microneedle every hour from the 512-cell to the sphere stage, and then developmental rates were evaluated at the hatching stage. The highest survival rate was obtained when we performed this manipulation at the 1k-cell stage. These results clearly showed that the MBT is the best stage for transplantation of PGCs or any cells in pikeperch. We described PGC migration and performed blastomere transplantation in pikeperch. PGCs were visualised by injection of synthesised green fluorescent protein (GFP) within the 3'untranslated region (UTR) mRNA of nanos3. GFP-positive PGCs appeared in all embryos at approximately 100% epiboly. Time-lapse imaging revealed the PGC migration pattern from their initial appearance to the location at the gonadal ridge. We conducted blastomere transplantation at the blastula stage. Donor embryos were labelled with GFP-nos3 3'UTR mRNA and tetramethylrhodamine dextran to label PGCs and somatic cells, respectively. Twelve blastomere transplantation chimeras were produced, with eight surviving to hatching. All exhibited donor-derived somatic cells in the developing body. The PGCs from donor embryos were observed to migrate towards the gonad region of the host embryos. Our results indicated that blastomere transplantation can be successfully applied in pikeperch, and these findings may be useful to produce germline chimeras in percids.