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Parallel single-cell analysis of active caspase-3/7 in apoptotic and non-apoptotic cells
Ledvina, Vojtěch ; Klepárník, Karel
Caspases are proteases that play key role in the process of apoptosis, the programmed\ncell death. Among them, caspase-3 and -7 are main executioner caspases that cleave\nmany vital proteins during apoptosis and after their widespread activation, the process\ncannot be reversed. To analyze caspase-3/7 activation within single cells, a miniaturized\ndevice for parallel analysis of eight samples was developed. The assay is based on the\nmodified luciferin-firefly luciferase bioluminescence (BL) system. Individual\nsuspended cells were collected and transferred into detection microvials using a\nmicromanipulator. The bioluminescence was detected using a photon counting head\nwith cooled photcathode. The LOD suitable for detection of active caspase-3/7 in both\napoptotic and non-apoptotic cells was reached.
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Neural Networks Between Integer and Rational Weights
Šíma, Jiří
The analysis of the computational power of neural networks with the weight parameters between integer and rational numbers is refined. We study an intermediate model of binary-state neural networks with integer weights, corresponding to finite automata, which is extended with an extra analog unit with rational weights, as already two additional analog units allow for Turing universality. We characterize the languages that are accepted by this model in terms of so-called cut languages which are combined in a certain way by usual string operations. We employ this characterization for proving that the languages accepted by neural networks with an analog unit are context-sensitive and we present an explicit example of such non-context-free languages. In addition, we formulate a sufficient condition when these networks accept only regular languages in terms of quasi-periodicity of parameters derived from their weights.
Fulltext: content.csg -  PDF
Plný tet: v1237-16 - PDF
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Therapeutic hypotermia after cardiac arrest
JIRCOVÁ, Danuše
The bachelor thesis deals with the influence of a mild therapeutic hypothermia as a hospital post-resuscitation care of patients after cardiopulmonary resuscitation for non-traumatic cardiac arrest. Therapeutic hypothermia is a relatively new method. It is part of the European Guidelines for resuscitation since 2005. It was reported its positive effects on the neurological status of the patients. Using the method reduces the damage to the brain and cardiac muscle. The theoretical part of the thesis deals with the effect of mild hypothermia on the human body. Specifies the indication and contraindication criteria for initiation of therapeutic hypothermia, describes cooling methods of the patient and ways to terminate the therapeutic hypothermia. Describes the monitoring of physiological functions and nursing care of patients. The practical part of bachelor thesis mapped the procedures used in the administration of therapeutic hypothermia in patients treated in two large hospitals. These were Fakultní nemocnice Kralovske Vinohrady in Prague and Nemocnice Ceske Budejovice, a.s. The applied strategy of the research was qualitative. Content analysis of 10 case reports were carried out, 5 case reports from each hospital. Case reports concerning patients after cardiopulmonary resuscitation, who were treated using the therapeutic hypothermia as a part of hospital care. The aim of this thesis was to map the procedures used in the therapeutic hypothermia administration in patients after cardiopulmonary resuscitation. It was made available by hospitals after approval by ethics committees on the basis of applications submitted. The information was subsequently created into case studies that were analysed in the categorization tables. In the discussion, the results were compared with the published knowledge provided in the theoretical part of the thesis, and were commented. Analysed were: the composition of the group of patients by age and gender, provide the basic life support by witnesses prior to arrival of crew of the emergency medical service on scene, initial ECG rhythm after emergency medical service arrival, the time elapsed since the cardiac arrest appeared to restore of spontaneous circulation, initiation of therapeutic hypothermia in pre-hospital care, time elapsed since the therapeutic hypothermia start up to reaching the target body temperature, duration of the therapeutic hypothermia, the methods used for cooling the patients, the methods used for measuring the body temperature and neurological status at the time of discharge or transfer from the hospital. The results of the research surprisingly showed that general public are well-informed. Witnesses were able to recognize cardiac arrest, early called in professional help and were willing to give heart massage. This shows also the success in implementing the method of medical dispatcher-assisted basic life support in practice, because the witnesses were in many cases instructed by the medical dispatchers to resuscitate the patient. The majority of patient's initial ECG rhythm were diagnosed as ventricular fibrillation. Time to restoration of spontaneous circulation in most cases ranged within 29 minutes. To achieve the target body temperature at the recommended time limit has managed to half of patients. Target body temperature was always kept within the recommended timeframe and core body temperature was monitored. Concerns raised by an absence of the method start up in pre-hospital emergency care. The thesis highlighted the seriousness of sudden cardiac arrest and the high incidence of patients dismissed from the department with severe neurological deficit. Therefore, it is necessary to continue in research of this method, improve it and try to incorporate it already to pre-hospital care.
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Effect of snowpack on runoff generation during rain on snow event.
Juras, Roman ; Máca, Petr (advisor) ; Ladislav , Ladislav (referee)
During a winter season, when snow covers the watershed, the frequency of rain-on-snow (ROS) events is still raising. ROS can cause severe natural hazards like floods or wet avalanches. Prediction of ROS effects is linked to better understanding of snowpack runoff dynamics and its composition. Deploying rainfall simulation together with hydrological tracers was tested as a convenient tool for this purpose. Overall 18 sprinkling experiments were conducted on snow featuring different initial conditions in mountainous regions over middle and western Europe.
Dye tracer brilliant blue (FCF) was used for flow regime determination, because it enables to visualise preferential paths and layers interface. Snowpack runoff composition was assessed by hydrograph separation method, which provided appropriate results with acceptable uncertainty. It was not possible to use concurrently these two techniques because of technical reasons, however it would extend our gained knowledge. Snowmelt water amount in the snowpack runoff was estimated by energy balance (EB) equation, which is very efficient but quality inputs demanding. This was also the reason, why EB was deployed within only single experiment.
Timing of snowpack runoff onset decrease mainly with the rain intensity. Initial snowpack properties like bulk density or wetness are less important for time of runoff generation compared to the rain intensity. On the other het when same rain intensity was applied, non-ripe snowpack featuring less bulk density created runoff faster than the ripe snowpack featuring higher bulk density. Snowpack runoff magnitude mainly depends on the snowpack initial saturation. Ripe snowpack with higher saturation enabled to generate higher cumulative runoff where contributed by max 50 %. In contrary, rainwater travelled through the non-ripe snowpack relatively fast and contributed runoff by approx. 80 %.
Runoff prediction was tested by deploying Richards equation included in SNOWPACK model. The model was modified using a dual-domain approach to better simulate snowpack runoff under preferential flow conditions. Presented approach demonstrated an improvement in all simulated aspects compared to the more traditional method when only matrix flow is considered.
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Design of Experiment for Non-Stationary Processes of Production
Jadrná, Monika ; Macák, Tomáš (advisor)
The doctoral thesis is concerned with the services sector and the area of mass production. Particularly, the optimization of the product portfolio of the travel agency and the optimization of production rounds of ammunition.
The theoretical part deals with the current overview of discussed topic. Further, the terminology and methods of the decision-making process are defined to support decision making. The theoretical basis of research focused on the choice of appropriate input variables in the area of services, and on the choice of a particular material option in the production area and appropriate equipment for the production. The theoretical part forms the basis for the practical part of the thesis.
For the doctoral thesis was chosen an enterprise operating in the defined sector. Product portfolio for the services sector is optimised using Fuzzy logic and Fuzzy sets so that the enterprise can maintain its competitiveness in todays highly ambitious market. Product portfolio for manufacture is optimised for achieving desired properties of the product.
The main aim of the thesis is to propose a new methodological approach for the management of selected business processes in their nonstationary time course. The aim of the practical implementation is to verify the functionality of the proposed methodological approach, both in the area of services and in the field of mass production.
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Population genetic analysis of Old Kladruber horse
Vostrá Vydrová, Hana ; Majzlík, Ivan (advisor) ; Karel, Karel (referee)
The Old Kladruber horse, along with the Lipizzaner horse, Andalusian horse and Lusitano horse, is of the original Italo-Spanish type. The Old Kladruber horse is kept in two colour varieties (grey and black). Because the population is closed, there is a concern about the loss of genetic variation. The genetic diversity and population structure were analysed in the Old Kladruber horse breed based on the pedigree information of animals that were registered in the Studbook to identify factors that may have affected the genetic variability of the breed. Pedigree records collected from 1729 to 2013 contained information on 7971 animals that were used in the analyses. The pedigree depth was up to 33 generations, with an average of 15.1 complete generations. The effective number of founders and ancestors contributing to the current genetic pool was 92.69 and 17.16, respectively. The average values of the inbreeding coefficient were as follows: 13% (with a maximum value of 29%) for the reference population (individuals that can currently take part in reproduction, n=612), 11% for the grey variety (with a maximum value of 25%) and 15% for the black variety (with a maximum value of 29%). The proportion of inbred animals was high (99%). The average rate of inbreeding in the reference population was 1%: 0.8% for the grey variety and 1.1% for the black variety, and the respective estimates of the effective population sizes were 52 for the reference population, 62 for the grey variety and 45 for the black variety. The estimated percentage of genetic diversity lost due to non-random mating within subpopulations and the reference population was 1.0, 1.0 and 1.2%, respectively. The total loss of genetic diversity in the reference population, in the grey variety and in the black variety was 11%, 13% and 17%, respectively.
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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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DCTOOL-A3
Bakule, Lubomír ; Papík, Martin ; Rehák, Branislav
DCTOOL-A3 is a documentation of Matlab routines developed for the design of decentralized control of large scale complex systems. The current beta version covers three areas as follows:\nReport 4.1 deals with the event-triggered control design for unstructured uncertain systems. Both non-quantized and quantized feedback is analyzed. The results are given in terms of linear matrix inequalities (LMIs). Logarithmic quantizer is used. Numerical example illustrates the effectiveness of the presented results.\nReport 4.2 presents a new decentralized overlapping wireless control design with a switched communication protocol. The method is applied by simulations on the 20-story building structure including the test of robustness of the methods against sensor failures and network node dropouts of a digital network.\nReport 4.3 presents the construction of a new decentralized wireless controller and a set of heuristic algorithms for evaluation of packet dropouts, sensor faults and actuator faults. The digital network operates at the standard frequency used in well-known widely-used industrial protocols. The results are tested at the Benchmark model decomposed into two disjoint substructures. The results are published. Thus, the details are omitted here.
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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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