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CE-nESI/MS in electrode-free design: narrowing the separation channel to 5 μm
Týčová, Anna ; Foret, František
Capillary electrophoresis-nanospray/mass spectrometry (CE-nESI/MS) in electrodefree\narrangement represents a very simple instrumentation for analysis of biomolecules.\nSo far, CE-nESI/MS analyses in this design were limited to 10-75 μm ID of the\nseparation channel. Although the work with narrower dimensions brings several\nchallenges, there is a great potential for better sensitivity, improved separation power\nand lower sample consumption. This work is devoted to some practical aspects of\nexperiments in narrow bore channels and systematic evaluation of CE-nESI/MS\nanalyses conducted in capillaries of 25, 15 and 5 μm ID.
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Performance of the Care Contribution Agenda in the Municipality with Extended Scope of Dačice after the Social Reform Seen by Recipients of the Care Contribution.
ČURDOVÁ, Hana
Since January 1, 2007 a new social security benefit has been provided: Care Allowance. This benefit is regulated by Act No. 108 of 2006, about social services. The Act defines conditions of help and benefit distribution to people, dependent on other person's help, who got into a difficult social situation. I have chosen the topic, regarding Care Allowance, on the basis of my professional experience in this issue. The aim of this thesis was to survey opinions of Care Allowance beneficiaries (families with a disabled child), regarding office workers' work before and after social reform and find out the impact of the reform on beneficiaries. The theoretical part contains a short reference to the situation before passing the Law of Social Services, a short introduction of the Law of Social Services (effective from January 1, 2007) and social reform. Next part of the thesis describes Care Allowance after social reform, thus after January 1, 2012. It specifically contains: Care Allowance impact on social services sphere, claim conditions, extend of dependance, benefit amount and its payment, Care Allowance proceedings, terms of authorities, Care Allowance use and duties resulting from the law. The thesis describes in detail a social worker, his or her role and authority. A term "handicap" was defined in the conclusion of the theorethical part. The practical part of the bachelor's thesis elicits communication partners' opinions on office workers' work. The research question was: How do Care Allowance beneficiaries ( families with a disabled child) evaluate office workers' work before and after social reform? On the basis of the research question, I set four questions and worked with them during the research. The research sample consisted of twelve families with a disabled child, who were chosen on the basis of their free decision. These families were Care Allowance beneficiaries before and after social reform, which is essential here. The research was carried out by means of quantitative survey, an inquiry method and a method of semi-structured dialogue with the parent with a disabled child, who draws Care Allowance. According to the facts elicited from the communication partners, social reform is perceived rather negatively than positively. There is a significant difference between work of social workers of municipal authorities in Dačice and work of social workers of employment office in Dačice. Communication partners see differences in helpfulness, satisfaction, knowledge and workers' experience. Employment office workers are evaluated by communication partners very negatively, on the contrary municipal authorities workers' work is evaluated absolutely positively. Communication partners mention another serious problem, which is misfunction of Care Allowance payment, the benefit is paid out late, up to one month later. In the conclusion of the thesis, I suggest solving to the problematic situation, that was revealed in the research.
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Analysis of organic additives in historical mortars
Křížová, Iva ; Čabala, Radomír (advisor) ; Hynek, Radovan (referee)
- 4 - Abstrakt The aim of this diploma thesis was to determine the possibility of identifying the protein and lipid additives, such as blood, gelatine, curd, lard and eggs, in the model samples of lime based mortars using Raman spectroscopy and gas chromatography- mass spectrometry. It was proved that the method of Raman spectroscopy can determine the presence of organic additives in the order of one weight percent. The gas chromatography enables to detect lipid additives from the order of hundredths weight percent. Unambiguous resolution of protein additives can be done in the samples with the concentrations up to the order of one weight percent of the additives.
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Determination of content substances for selected species and varieties of roses.
Slavíková, Eva ; Sus, Josef (advisor) ; Súkeníková, Tereza (referee)
This bachelor's work is focused on comparing the most significant content substance, vitamin C, in fruits of chosen rose kinds species and varieties. Herbal products are often seen by people as products of "second choice", when failure of allopathic treatment occurs, which is focused especially on suppressing symptoms of illness, but it ignores health of the human body as whole. That's why are herbal products often alternative for therapy of chronic diseases. The base of this work is extensive literary research and results of laboratory measurings. Literary part of this work includes description of botanical classification of rose plants. There is stated basic characteristic of individual rose species and their morphological features. In following chapters is composed overview of effective substances in rose fruits and their effect on human health. In practical part of this work are included results of laboratory tests for determining ascorbic acid content for chosen samples of rose species and it also includes results of weighing and measuring of fruit samples of chosen rose species. There are also stated chosen rose species and shortly their origin and description, in the beginning of practical part. For practical part were chosen samples of species. Results of practical part are compiled into statistic tables and graphs and it confirms high content of ascorbic acid in rose fruits. This content is in range of cca 300 - 900 mg.100 g-1. Due to growing interest in alternative treatment options and "clean" natural product, it can be assumed, that many plants of Rosaceae family will keep its place in treatment practice. Those plants will be used not only for treatment of acute and chronic diseases, but also for cosmetic products production, thanks to its large effects and process options.
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Nutritional analysis and optimization of breeding of selected species of edible insects under conditions of the Czech Republic with regard to human health
Adámková, Anna ; Kouřimská, Lenka (advisor)
All over the world, the edible insects are considered a highly nutritious food with high protein and fat content. However, the nutritional value of insect is not constant. It can be affected by species, developmental stage, rearing technology or nutrition.
Therefore, this thesis was aimed at obtaining the selected nutritional value of edible insects. Analyses were focused on the determination of the crude protein content, fat content, fatty acid profile and sterols in selected species of edible insects, which can be commonly reared in the Czech Republic. At the same time we also analysed samples of insects reared on the island of Sumatra to evaluate the influence of the climate on the nutritional value. The main aim of the thesis was the determination of optimal breeding conditions, developmental stages and feed rations for obtaining good production of insects with nutritional properties suitable for human nutrition. The analyses showed a high nutritional value of selected insect species, but also confirmed the significant differences in the content of individual nutrients between different species depending on the climate conditions and developmental stage. Comparing the fat and crude protein content in edible insects and other conventional sources of meat it has been found, that the examined insect is similar to beef concerning the fat and crude protein content. The results obtained are the basis for determining the appropriate rearing conditions and developmental stages for obtaining insect with the desired nutritional properties for human nutrition.
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Freezing technology of bull sperm in relation to its survivability and fertilization ability
Doležalová, Martina ; Stádník, Luděk (advisor) ; Jiří, Jiří (referee)
The aim of optimalization the insemination doses production is to provide the highest fertilization ability of spermatozoa during the demanding proces of processing fresh semen and its subsequent cryopreservation. Temperature changes causes spermatozoa damage during the cooling and freezing. Spermatozoa is exposed to cold shock and many others limiting factors, which leads to cell death and therefore to decline of fertilization ability of thawed insemination doses. For increasing spermatozoa resistance, exactly the plasma membrane resistance against cold shock was fraction of egg yolk LDL cholesterol (low density lipoprotein) at various concentrations into the comercially produced diluents added. It is believed that LDL acts possitively to plasma membrane and helps to maintain the fertilization ability of spermatozoa after thawing. Following step in the proces of insemination doses production is slow cooling of diluted semen and equilibration, when the straws are store at cooling box for 30 minutes to 240 hours. This period is necessary to penetrate of certain diluent components into the spermatazoa also maintain the balance between their intracellular and extracellular concentration. Also important is subsequent freezing temperature gradient of insemination doses. The most suitable freezing method is based on computer controlled temperature decline in freezing chamber which allows the precise control of ice crystals formation that could tear and kill the cell.
During 2012 to 2016 was repeatedly collected semen from the group of breeding bulls (n = 27, Holstein and Czech Fleckvieh breed) at AI centre. Semen which fulfill the standard entrance conditions in first step was evenly into several parts divided. For dilution the three types of comercially diluents AndroMed, Bioxcell and Triladyl with and without LDL addition were used. Into the diluents AndroMed and Bioxcell the concentration of LDL 4 %, 6 % and 8% into the dilent Triladyl 6 %, 8 % and 10 % was added. Diluted semen was filled into the glass capillares with volume 0,1 ml and temperature +4 °C. Subsequently the sample was placed to cold bath (0°C) for 10 minutes. Then the volume of capillare with physiological solution (37 °C) was mixed and for next 120 minutes was incubate. The effect of cold shock to proportion of live spermatozoa was evaluated by using Eosin and Nigrosine staining technique during heat test of spermatozoa survivability after spermatozoa heating and after 120 minutes of incubation.
The more suitable semen diluents which provide the higher spermatozoa resistance against cold shock were AndroMed and Bioxcell. Together the possitive effect of LDL addition into the diluents to lower decrease of proportion of live spermatozoa during heat test was found (P<0.05). The most suitable LDL concentration which had a favorable influence at spermatozoa resistance against cold shock was 6 % in diluent Bioxcell. Values of the proportion of live sperm were higher at the beginning of the heat test (+1.31% to + 3.2%) and after 120 minute incubation (+5.82% to +8.41%) compared to other diluents with and without addition of LDL.
In the next step the process of equilibration was optimized, is an important part of insemination doses production. The effect of the length of equilibration for subsequent fertilization ability of spermatozoa was evaluated using spermatozoa motility based of CASA and proportion of live spermatozoa after thawing and during heat survival test lasting 120 minutes (37 ° C). Suitable semen was diluted by comercially used diluent AndroMed based on soya lecithin, filled into the straws (0.25 ml), cooled and equilibrated in cooling box for 30, 120 and 240 minutes and freezed in programmable freezing box applying four types of freezing curves differing in temperature rate decline. There was used standard and by producer recommended 3. phase freezing curve, then 2. phase freezing curve, and 3. phase freezing curve with slower as well as rapid decline of temperature rate in freezing chamber, compared with standard freezing curve. The highest spermatozoa motility was found using 240 minutes of equilibration by +2.72% and +4.58% compared to other lengths of equilibration (P <0.05 to 0.01). The highest proportion of live spermatozoa was found using 120 minutes of equilibration (+6.87 % and +8.68 %). The highest average spermatozoa motility during heat test after thawing was achieved by using 2. phase freezing curve (from +2.97% to +10.37%, P <0.05), also in the proportion of live spermatozoa (from + 4.37% to +8.82%, P <0.01). When evaluating interaction between the length of equilibration and freezing curve (standard 3. phase and 2 . phase freezing curve), the highest average spermatozoa motility and proportion of live spermatozoa using 240 minutes of equilibration by both freezing curves was reached, there was no statistically significant differences. As well as, in all evaluated parts of this study the individual differences between ejaculate of bulls and within semen from one bull (P <0.05) as secondary effect were found.
To maintain good fertilization ability of semen during cryopreservation is necessary to increase the spermatozoa resistance against cold shock using addition of correct concentration of LDL into the commercially used diluents AndroMed and Bioxcell. Subsequently the fertilization ability of insemination dose is influenced by cooling, the length of equilibration and freezing. The length of equilibration 120 minutes and more as well as gentle way of freezing according to freezing curve, which ensures a gradual decrease of temperature in freezing chamber provided the higher average spermatozoa motility and proportion of live spermatozoa.
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Antioxidant, anti-proliferative and immunomodulatory effect of fruits, vegetables and mushrooms in vitro
Doskočil, Ivo ; Havlík, Jaroslav (advisor) ; Orsák, Matyáš (referee)
Recently there has been an increasing interest in discovering of new species of plants and mushrooms which have antioxidant or anti proliferative activity. The interest is caused by the fact that these species have medicinal and food utilization. These properties of plants and mushrooms can be used when dealing with many diseases which may be connected with oxidative stress (inflammatory bowel disease, cardiovascular disease, hypertension, and tumour disease, etc.).
The thesis propounds characteristics of in vitro antioxidant and anti proliferative activity of 19 types of juices and 28 methanol extracts of fruits and vegetables, which are common parts of a diet. In the thesis there are also characteristics of 13 ethanol extracts of edible mushrooms of the order of Polypore (Popyporales). Antioxidant activity was quantifie by 2, 2 diphenyl 1 picrylhydrazyl (DPPH), oxygen radical absorbance capacity (ORAC), and inhibition of nitric oxide (NO) production. Cytotoxicity was measured by MTT (3 (4,5 dimethylthiazol 2 yl) 2,5 diphenyltetrazolium bromide. Immunomodulatory activity was determined by an assay of phagocytic activity of human neutrophil granulocytes. Further a total phenolic content (TPC) and total beta glucan content were investigated; these are considered to be the potentially active constituents participating in mentioned activities.
From the obtained results it is evident that juices (TPC = 1603.2 mg GAE/L; ORAC = 438.5 umol TE/g)
and a bilberry (Vaccinium myrtillus) fruit extract (ORAC 836.6 umol TE/g; DPPH = 404.6 umol TE/g) showed
the highest antioxidant activity of all tested samples of fruits and vegetables. Capsicum (Capsicum L.)
juices (TPC = 642.1 mg GAE/L; ORAC = 127.9 umol TE/g) and a radish (Raphanus sativus L.) extract
(ORAC 724.5 umol TE/g; DPPH = 52.2 umol TE/g) also proved to have the high values of antioxidant activity. From the tested fruits and vegetables the following inhibited to produce a nitric oxide: onion (Allium cepa L) juices (lowered the NO production by 57%), tangerine juices (Citrus reticulata Blanco)(by 52%), broccoli (Brassica oleracea var. botrytis italic) extract (by 21%), and orange (Citrus sinensis Pers.) extract (by 10%). Concerning edible
mushrooms, Lentinus tigrinus (Bull.) Fr. (TPC = 216.2 umol GAE/g of extract), Ganoderma lucidum (Curtis) P. Kras (TPC = 257.9 umol GAE/g of extract), and Royoporus badius (Pers.) A.B. De (TPC = 257.8 umol GAE/g of extract) presented the highest phenolic content. Sparassis crispa (Wulfen) Fr. (117.4 mg/g of extract) had the highest content of beta glucan. Substantial effect on phagocytic activity of granulocytes was noticed in connection with Neolentinus lepideus (Fr.) Redhead & Ginns, Polyporus squamosus (Huds.) Fr., and S. crispa. In connection with the latter of mentioned mushrooms, a moderate inhibitory effect towards HT 29 human colorectal adenocarcinoma cell line
(IC50 = 107 ug/ml of extract) was noticed.
The results indicate that the tested plants and mushrooms can be beneficial when reducing the negative effects of oxidative stress. The oxidative stress has been recognized as a contributing factor to a whole range of diseases and the reduction of the oxidative stress may lead to the decrease in the possibility of the diseases´ progression. The results show the possible favourable effects on the human health. When evaluating the results, the following fact should be taken into consideration; the in vitro tests and screenings are considered to be the first phase of systematic research of the effects and serve for the choice of respondents for further detailed studies.
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Non-invasive methods for sex and steroid homones determination in gibbons of the genus Nomascus
Bolechová, Petra ; Bartoš, Luděk (advisor) ; Pavel, Pavel (referee)
The gibbon primates of the family Nomascus are classified as critically endangered species, and, to date, basic understanding and information about their biology is missing. With regard to the status of these animals in the wild and captive populations in zoos, being familiar with their reproduction, may improve captive breeding programs. Data collection in this study was to be carried out by the practical use of non-invasive methods (polymerase chain reaction for DNA extraction and enzyme immunoassays), using faecal samples for analysis. The first method, polymerase chain reaction, was used for sex determination in juveniles, because of their coat colour and visual similarity of secondary sex characteristic in both sexes; it is not possible to determinate sex without handling the animal. Another main purpose of this study is to try and answer the hypothesis regarding the ovarian cycle of females, factors influencing their hormone concentration and also the onset of sexual maturity in females and the timing of their fur colour change. Hypotheses were checked by monitoring the concentration of progesterone and oestrogen faecal metabolites and by evaluation of the composition of breeding groups of gibbons with the ZOO influence. During a four year period (from 2010 till 2014), there were a total of 51 animals analysed from 16 different zoos with a faecal sample count totalling 1618 samples. The results confirmed the use of noninvasive methods for sex determination, and thus ensuring the maximum welfare standards. Endocrinological analysis confirmed the hypothesis of the influence of the environment (ZOO) to the hormone concentrations and female´s pregnancy effect in conjunction with her age on the final results of the male - father hormone concentrations. The initial information in this study is the confirmation of the onset of ovarian cycle in young females without connection to their fur colour change and a significant factor of a mother´s impact and her territoriality. This study is the first to present a long term monitoring of ovarian cycles in females and hormone concentrations of other individuals, both male and female, and from various age groups. The results allow us to understand the possible impact of zoo environments on the reproductive status of these gibbons and contribute to the general improvement of breeding management.
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Time lapse tomography of fracture progress in silicate-based composite subjected to the loading a combination with acoustic emission scanning
Kumpová, Ivana ; Kytýř, Daniel ; Fíla, Tomáš ; Veselý, V. ; Trčka, T. ; Vopálenský, Michal ; Vavřík, Daniel
The initiation and propagation of a fracture in quasi-brittle materials (such as silicatebased composite) is an increasingly discussed topic for which various methods of research have been developed/applied. As the quasi-brittle silicate-based composite compounds are very non-homogenous, the mechanism of the crack initiation and propagation can be very different even for samples with the\nidentical geometry. One possible approach to study the fracture mechanism in quasi-brittle materials is to use several different experimental techniques in a single experiment and perform detail analysis to identify generally valid fracture process phenomena. In this work, a simultaneous monitoring of fracture\nprocess zone formation and propagation by three different methods is presented and discussed. A three point bending test was performed on a notched silicate composite specimen. During the loading process, a highly accurate force displacement dependence was recorded accompanied with X-ray radiography,\nX-ray computed tomogra-phy and acoustic emission scanning.
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The impact analysis of the technical devices used by the driver on the reaction time
Kubánek, Ondřej ; Bradáč, Albert (referee) ; Panáček, Vladimír (advisor)
This master‘s thesis analyzes the impact of the use of technical equipment for the driver reaction time. There are described the legal requirements for the use of technical devices by the driver in the vehicle. There are analyzed the types of technical devices used by drivers and mainly discussed the issue of drivers' reaction times. They assessed factors affecting reaction time, ways of measuring the reaction time and the type of reaction times. They discussed earlier research from the beginning of the measurement of reaction times to the latest research, dealing with the measurement of reaction times when using a mobile phone. The practical part is devoted to perform road tests on a sample of drivers and evaluate their reaction times during normal driving and during a call. The measured and evaluated data are commented with respect to the benefits of expert activities in the field of transport.
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