Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 20

Search results for: Zuzana Sutova

20 Inverse Matrix in the Theory of Dynamical Systems

Authors: Renata Masarova, Bohuslava Juhasova, Martin Juhas, Zuzana Sutova

Abstract:

In dynamic system theory a mathematical model is often used to describe their properties. In order to find a transfer matrix of a dynamic system we need to calculate an inverse matrix. The paper contains the fusion of the classical theory and the procedures used in the theory of automated control for calculating the inverse matrix. The final part of the paper models the given problem by the Matlab.

Keywords: dynamic system, transfer matrix, inverse matrix, modeling

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19 Tool for Analysing the Sensitivity and Tolerance of Mechatronic Systems in Matlab GUI

Authors: Bohuslava Juhasova, Martin Juhas, Renata Masarova, Zuzana Sutova

Abstract:

The article deals with the tool in Matlab GUI form that is designed to analyse a mechatronic system sensitivity and tolerance. In the analysed mechatronic system, a torque is transferred from the drive to the load through a coupling containing flexible elements. Different methods of control system design are used. The classic form of the feedback control is proposed using Naslin method, modulus optimum criterion and inverse dynamics method. The cascade form of the control is proposed based on combination of modulus optimum criterion and symmetric optimum criterion. The sensitivity is analysed on the basis of absolute and relative sensitivity of system function to the change of chosen parameter value of the mechatronic system, as well as the control subsystem. The tolerance is analysed in the form of determining the range of allowed relative changes of selected system parameters in the field of system stability. The tool allows to analyse an influence of torsion stiffness, torsion damping, inertia moments of the motor and the load and controller(s) parameters. The sensitivity and tolerance are monitored in terms of the impact of parameter change on the response in the form of system step response and system frequency-response logarithmic characteristics. The Symbolic Math Toolbox for expression of the final shape of analysed system functions was used. The sensitivity and tolerance are graphically represented as 2D graph of sensitivity or tolerance of the system function and 3D/2D static/interactive graph of step/frequency response.

Keywords: mechatronic systems, Matlab GUI, sensitivity, tolerance

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18 Nanoparticles of Hyaluronic Acid for Radiation Induced Lung Damages

Authors: Anna Lierova, Jitka Kasparova, Marcela Jelicova, Lucie Korecka, Zuzana Bilkova, Zuzana Sinkorova

Abstract:

Hyaluronic acid (HA) is a simple linear, unbranched polysaccharide with a lot of exceptional physiological and chemical properties such as high biocompatibility and biodegradability, strong hydration and viscoelasticity that depend on the size of the molecule. It plays the important role in a variety of molecular events as tissue hydration, mechanical protection of tissues and as well as during inflammation, leukocyte migration, and extracellular matrix remodeling. Also, HA-based biomaterials, including HA scaffolds, hydrogels, thin membranes, matrix grafts or nanoparticles are widely use in various biomedical applications. Our goal is to determine the radioprotective effect of hyaluronic acid nanoparticles (HA NPs). We are investigating effect of ionizing radiation on stability of HA NPs, in vitro relative toxicity of nanoscale as well as effect on cell lines and specific surface receptors and their response to ionizing radiation. An exposure to ionizing radiation (IR) can irreversibly damage various cell types and may thus have implications for the level of the whole tissue. Characteristic manifestations are formation of over-granulated tissue, remodeling of extracellular matrix (ECM) and abortive wound healing. Damages are caused by either direct interaction with DNA and IR proteins or indirectly by radicals formed during radiolysis of water Accumulation and turnover of ECM are a hallmark of radiation induces lung injury, characterized by inflammation, repair or remodeling health pulmonary tissue. HA is a major component of ECM in lung and plays an important role in regulating tissue injury, accelerating tissue repair, and controlling disease outcomes. Due to that, HA NPs were applied to in vivo model (C57Bl/6J mice) before total body or partial thorax irradiation. This part of our research is targeting on effect of exogenous HA on the development and/or mitigating acute radiation syndrome and radiation induced lung injuries.

Keywords: hyaluronic acid, ionizing radiation, nanoparticles, radiation induces lung damages

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17 Change of Internal Friction on Magnesium Alloy with 5.48% Al Dependence on the Temperature

Authors: Milan Uhríčik, Andrea Soviarová, Zuzana Dresslerová, Peter Palček, Alan Vaško

Abstract:

The article is focused on the analysis changes dependence on the temperature on the magnesium alloy with 5,48% Al, 0,813% Zn and 0,398% Mn by internal friction. Internal friction is a property of the material is measured on the ultrasonic resonant aparature at a frequency about f = 20470 Hz. The measured temperature range was from 30 °C up to 420 °C. Precisely measurement of the internal friction can be monitored ongoing structural changes and various mechanisms that prevent these changes.

Keywords: internal friction, magnesium alloy, temperature, resonant frequency

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16 Support of Knowledge Sharing in Manufacturing Companies: A Case Study

Authors: Zuzana Crhová, Karel Kolman, Drahomíra Pavelková

Abstract:

Knowledge is considered as an important asset which can help organizations to create competitive advantage. The necessity of taking care of these assets is more important in these days – in days of turbulent changes in business environment. Knowledge could facilitate adaption to constant changes. The aim of this paper is to describe how the knowledge sharing can be supported in the manufacturing companies. The methods of case studies and grounded theory were used to present information gained by carrying out semi-structured interviews. Results show that knowledge sharing is supported in very similar ways in respondent companies.

Keywords: case study, human resource management, knowledge, knowledge sharing

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15 The Use of Methods and Techniques of Drama Education with Kindergarten Teachers

Authors: Vladimira Hornackova, Jana Kottasova, Zuzana Vanova, Anna Jungrova

Abstract:

Present study deals with drama education in preschool education. The research made in this field brings a qualitative comparative survey with the aim to find out the use of methods and techniques of drama education in preschool education at university or secondary school graduate preschool teachers. The research uses a content analysis and an unstandardized questionnaire for preschool teachers and obtained data are processed with the help of descriptive methods and correlations. The results allow a comparison of aspects applied through drama in preschool education. The research brings impulses for education improvement in kindergartens and inspiration for university study programs of drama education in the professional training of preschool teachers.

Keywords: drama education, preschool education, preschool teacher, research

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14 DNA Methylation Changes Caused by Lawsone

Authors: Zuzana Poborilova, Anna B. Ohlsson, Torkel Berglund, Anna Vildova, Petr Babula

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Lawsone is a pigment that occurs naturally in plants. It has been used as a skin and hair dye for a long time. Moreover, its different biological activities have been reported. The present study focused on the effect of lawsone on a plant cell model represented by tobacco BY-2 cell suspension culture, which is used as a model comparable with the HeLa cells. It has been shown that lawsone inhibits the cell growth in the concentration-dependent manner. In addition, changes in DNA methylation level have been determined. We observed decreasing level of DNA methylation in the presence of increasing concentrations of lawsone. These results were accompanied with overproduction of reactive oxygen species (ROS). Since epigenetic modifications can be caused by different stress factors, there could be a connection between the changes in the level of DNA methylation and ROS production caused by lawsone.

Keywords: DNA methylation, lawsone, naphthoquinone, reactive oxygen species

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13 The Use of Emoticons in Polite Phrases of Greeting and Thanks

Authors: Zuzana Komrsková

Abstract:

This paper shows the connection between emoticons and politeness in written computer-mediated communication. It studies if there are some differences in the use of emoticon between Czech and English written tweets. My assumptions about the use of emoticons were based on the use of greetings and thanks in real, face to face situations. The first assumption, that welcome greeting phrase would be accompanied by positive emoticon was correct. But for the farewell greeting both positive and negative emoticons are possible. My results show lower frequency of negative emoticons in this context. I also found quite often both positive and negative emoticon in the same tweet. The expression of gratitude is associated with positive emotions. The results show that emoticons accompany polite phrases of greeting and thanks very often both in Czech and English. The use of emoticons with studied polite phrases shows that emoticons have become an integral part of these phrases.

Keywords: Czech, emoticon, english, politeness, twitter

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12 On Tarski’s Type Theorems for L-Fuzzy Isotone and L-Fuzzy Relatively Isotone Maps on L-Complete Propelattices

Authors: František Včelař, Zuzana Pátíková

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Recently a new type of very general relational structures, the so called (L-)complete propelattices, was introduced. These significantly generalize complete lattices and completely lattice L-ordered sets, because they do not assume the technically very strong property of transitivity. For these structures also the main part of the original Tarski’s fixed point theorem holds for (L-fuzzy) isotone maps, i.e., the part which concerns the existence of fixed points and the structure of their set. In this paper, fundamental properties of (L-)complete propelattices are recalled and the so called L-fuzzy relatively isotone maps are introduced. For these maps it is proved that they also have fixed points in L-complete propelattices, even if their set does not have to be of an awaited analogous structure of a complete propelattice.

Keywords: fixed point, L-complete propelattice, L-fuzzy (relatively) isotone map, residuated lattice, transitivity

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11 Gas Separation by Water-Swollen Membrane

Authors: Lenka Morávková, Zuzana Sedláková, Jiří Vejražka, Věra Jandová, Pavel Izák

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The need to minimize the costs of biogas upgrading leads to a continuous search for new and more effective membrane materials. The improvement of biogas combustion efficiency is connected with polar gases removal from a feed stream. One of the possibilities is the use of water–swollen polyamide layer of thin film composite reverse osmosis membrane for simultaneous carbon dioxide and hydrogen sulphide removal. Transport properties and basic characteristics of a thin film composite membrane were compared in the term of appropriate water-swollen membrane choice for biogas upgrading. SEM analysis showed that the surface of the best performing composites changed significantly upon swelling by water. The surface changes were found to be a proof that the selective skin polyamide layer was swollen well. Further, the presence of a sufficient number of associative centers, namely amido groups, inside the upper layer of the hydrophilic thin composite membrane can play an important role in the polar gas separation from a non-polar gas. The next key factor is a high porosity of the membrane support.

Keywords: biogas upgrading, carbon dioxide separation, hydrogen sulphide separation, water-swollen membrane

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10 Ionic Liquid Membranes for CO2 Separation

Authors: Zuzana Sedláková, Magda Kárászová, Jiří Vejražka, Lenka Morávková, Pavel Izák

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Membrane separations are mentioned frequently as a possibility for CO2 capture. Selectivity of ionic liquid membranes is strongly determined by different solubility of separated gases in ionic liquids. The solubility of separated gases usually varies over an order of magnitude, differently from diffusivity of gases in ionic liquids, which is usually of the same order of magnitude for different gases. The present work evaluates the selection of an appropriate ionic liquid for the selective membrane preparation based on the gas solubility in an ionic liquid. The current state of the art of CO2 capture patents and technologies based on the membrane separations was considered. An overview is given of the discussed transport mechanisms. Ionic liquids seem to be promising candidates thanks to their tunable properties, wide liquid range, reasonable thermal stability, and negligible vapor pressure. However, the uses of supported liquid membranes are limited by their relatively short lifetime from the industrial point of view. On the other hand, ionic liquids could overcome these problems due to their negligible vapor pressure and their tunable properties by adequate selection of the cation and anion.

Keywords: biogas upgrading, carbon dioxide separation, ionic liquid membrane, transport properties

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9 Determination of Prostate Specific Membrane Antigen (PSMA) Based on Combination of Nanocomposite [email protected]@JB303 and Magnetically Assisted Surface Enhanced Raman Spectroscopy (MA-SERS)

Authors: Zuzana Chaloupková, Zdeňka Marková, Václav Ranc, Radek Zbořil

Abstract:

Prostate cancer is now one of the most serious oncological diseases in men with an incidence higher than that of all other solid tumors combined. Diagnosis of prostate cancer usually involves detection of related genes or detection of marker proteins, such as PSA. One of the new potential markers is PSMA (prostate specific membrane antigen). PSMA is a unique membrane bound glycoprotein, which is considerably overexpressed on prostate cancer as well as neovasculature of most of the solid tumors. Commonly applied methods for a detection of proteins include techniques based on immunochemical approaches, including ELISA and RIA. Magnetically assisted surface enhanced Raman spectroscopy (MA-SERS) can be considered as an interesting alternative to generally accepted approaches. This work describes a utilization of MA-SERS in a detection of PSMA in human blood. This analytical platform is based on magnetic nanocomposites [email protected], functionalized by a low-molecular selector labeled as JB303. The system allows isolating the marker from the complex sample using application of magnetic force. Detection of PSMA is than performed by SERS effect given by a presence of silver nanoparticles. This system allowed us to analyze PSMA in clinical samples with limits of detection lower than 1 ng/mL.

Keywords: diagnosis, cancer, PSMA, MA-SERS, Ag nanoparticles

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8 Current Approach in Biodosimetry: Electrochemical Detection of DNA Damage

Authors: Marcela Jelicova, Anna Lierova, Zuzana Sinkorova, Radovan Metelka

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At present, electrochemical methods are used in various research fields, especially for analysis of biological molecules. The fact offers the possibility of using the detection of oxidative damage induced indirectly by γ rays in DNA in biodosimentry. The main goal of our study is to optimize the detection of 8-hydroxyguanine by differential pulse voltammetry. The level of this stable and specific indicator of DNA damage could be determined in DNA isolated from peripheral blood lymphocytes, plasma or urine of irradiated individuals. Screen-printed carbon electrodes modified with carboxy-functionalized multi-walled carbon nanotubes were utilized for highly sensitive electrochemical detection of 8-hydroxyguanine. Electrochemical oxidation of 8-hydroxoguanine monitored by differential pulse voltammetry was found pH-dependent and the most intensive signal was recorded at pH 7. After recalculating the current density, several times higher sensitivity was attained in comparison with already published results, which were obtained using screen-printed carbon electrodes with unmodified carbon ink. Subsequently, the modified electrochemical technique was used for the detection of 8-hydroxoguanine in calf thymus DNA samples irradiated by 60Co gamma source in the dose range from 0.5 to 20 Gy using by various types of sample pretreatment and measurement conditions. This method could serve for fast retrospective quantification of absorbed dose in cases of accidental exposure to ionizing radiation and may play an important role in biodosimetry.

Keywords: biodosimetry, electrochemical detection, voltametry, 8-hydroxyguanine

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7 Structural and Magnetic Properties of CoFe2O4:Nd3+/Dy3+/Pr3+/Gd3+ Nanoparticles Synthesized by Starch-Assisted Sol-Gel Auto-Combustion Method and Annealing Effect

Authors: Raghvendra Singh Yadav, Ivo Kuřitka, Jaromir Havlica, Zuzana Kozakova, Jiri Masilko, Lukas Kalina, Miroslava Hajdúchová, Vojtěch Enev, Jaromir Wasserbauer

Abstract:

In this work, we investigated the structural and magnetic properties of CoFe2O4:Nd3+/Dy3+/Pr3+/Gd3+ nanoparticles synthesized by starch-assisted sol-gel combustion method. X-ray diffraction pattern confirmed the formation of cubic spinel structure of rare-earth ions (Nd3+, Dy3+, Pr3+, Gd3+) doped CoFe2O4 spinel ferrite nanoparticles. Raman and Fourier Transform Infrared spectroscopy study also confirmed cubic spinel structure of rare-earth ions (Nd3+, Dy3+, Pr3+, Gd3+) substituted CoFe2O4 nanoparticles. The field emission scanning electron microscopy study revealed the effect of annealing temperature on size of rare-earth ions (Nd3+, Dy3+, Pr3+, Gd3+) substituted CoFe2O4 nanoparticles and particles were in the range of 10-100 nm. The magnetic properties of rare-earth ions (Nd3+, Dy3+, Pr3+, Gd3+) substituted CoFe2O4 nanoparticles were investigated by using vibrating sample magnetometer. The variation in saturation magnetization, coercivity and remanent magnetization with annealing temperature/ particle size of rare-earth ions (Nd3+, Dy3+, Pr3+, Gd3+) substituted CoFe2O4 nanoparticles was observed. Acknowledgment: This work was supported by the Ministry of Education, Youth and Sports of the Czech Republic – Program NPU I (LO1504).

Keywords: starch, sol-gel combustion method, rare-earth ions, spinel ferrite nanoparticles, magnetic properties

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6 Patient Advocates to Improve Access to Justice in Involuntary Hospitalisation

Authors: Zuzana Durajova, Natasa Diatkova, Shreya Bhardwaj

Abstract:

This paper introduces the project START, its activities, goals, evaluation and final results. Over the past few decades, the legal discourse surrounding mental health has resulted in improvement in patient rights (in Netherlands, etc.), the appointment of Ombudspersons for psychiatric patients (in Austria, Sweden) and facilitating the participation of patients in decision-making processes. Czech legislation already recognizes the position of “patient’s advocate” as a person of trust. However, this instrument is not very widely known and rarely used in practice. In the pilot study of the project, legal training for patient advocacy is provided to persons with experience with mental health problems/psychiatric hospitalization chosen from a Czech-based NGO. These persons (patient advocates) visit patients in involuntary hospitalization in one closed ward in the chosen psychiatric institution. During visits, the patient advocates inform patients about their legal standing, their procedural rights and also offer them individual support in contacting their counsel, family members etc. To understand the effect of the intervention, qualitative interviews and participant observations are conducted with the patients, advocates, the hospital management and staff and other identifiable stakeholders, such as government officials responsible for mental health care reform. The interviews are held before, during and after the intervention (support from patient advocates in hospitals). Given the ethical quandaries arising from using psychiatric wards as a field setting, we assume a participatory approach to ensure respect for patient boundaries and dignity. Through this project, we seek to establish a profession of patient advocates based on professional standards.

Keywords: patient advocacy, involuntary hospitalization, Czech Republic, patient Rights, professionalization

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5 Characterization of Cyanobacteria as Polyhydroxybutyrate Producer

Authors: Eva Slaninova, Diana Cernayova, Zuzana Sedrlova, Katerina Mrazova, Petr Sedlacek, Jana Nebesarova, Stanislav Obruca

Abstract:

Cyanobacteria are Gram-negative prokaryotes that belong to a group of photosynthetic bacteria. In comparison with heterotrophic microorganisms, cyanobacteria utilize atmospheric nitrogen and carbon dioxide without any additional substrates. This ability of these microorganisms could be employed in biotechnology for the production of bioplastics, concretely polyhydroxyalkanoates (PHAs) which are primarily accumulated as a storage material in cells in the form of intracellular granules. In this study, two cyanobacterial cultures from genera Synechocystis were used, namely Synechocystic sp. PCC 6803 and Synechocystis salina CCALA 192. There were optimized and used several various approaches, including microscopic techniques such as Cryo-scanning electron microscopy (Cryo-SEM) and transmission electron microscopy (TEM) and fluorescence lifetime imaging microscopy using Nile red as a fluorescent probe (FLIM). Due to these instrumental techniques, the morphology of intracellular space and surface of cells were characterized. The next group of methods that were employed was spectroscopic techniques such as UV-Vis spectroscopy measured in two modes (turbidimetry and integrating sphere) and Fourier transform infrared spectroscopy (FTIR). All these diverse techniques were used for detection and characterization of pigments (chlorophylls, carotenoids, phycocyanin, etc.) and PHAs, in our case poly(3-hydroxybutyrate) (P3HB). To verify results, gas chromatography (GC) was employed concretely for the determination of the amount of P3HB in biomass. Cyanobacteria were also characterized as polyhydroxybutyrate producers by flow cytometer, which could count cells and at the same time distinguish cells including P3HB and without P3HB due to fluorescent probe called BODIPY and live/dead fluorescent probe SYTO Blue. Based on the results, P3HB content in cyanobacteria cells was determined and also overall fitness of the cells. Funding: This study was partly funded by the project GA19- 19-29651L of the Czech Science Foundation (GACR) and partly funded by the Austrian Science Fund (FWF), a project I 4082-B25.

Keywords: Cyanobacteria, fluorescent probe, microscopic techniques, poly(3-hydroxybutyrate), chromatography, spectroscopy

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4 The Indebtedness of Men and Women: A Study of Personal Bankruptcies in the Czech Republic

Authors: Zuzana Fišerová, Marie Paseková

Abstract:

Debt relief (also labelled personal bankruptcy) is a bankruptcy settlement method which was implemented into Czech legislation by the Insolvency Act (Act No. 182/2006 Coll. on Insolvency and its Resolution) on 1 January 2008. The need to implement the institute of personal bankruptcy arose from the excessive over-indebtedness of many inhabitants of the Czech Republic after the crisis that arose around 2008 and 2009. The contribution analyses the development in the manner in which households approach personal bankruptcy and assesses and surveys the differences between indebtedness among men and women. The first section analyses the development in numbers of filed personal bankruptcy petitions and the successfulness thereof; it likewise analyses the impact of other economic influences (regional differences, unemployment etc.). The differences between debtors in dependency to gender are also surveyed. A survey of insolvency proceedings for 664 persons whose insolvency proceedings were commenced in 2008 was conducted, whilst the data were acquired from the publicly accessible insolvency register. The hypothesis on the equality of the average debt level of men and women was tested when comparing indebtedness in dependency to debtor gender. At a significance level of 0.05, the test confirmed that the mean value of debt level for women is lower than the mean value of debt level for men. Through analysis of further results, it was found that the average level of debt among women was CZK 537 thousand, while the average level of creditor satisfaction reached 46.2%. Men in the monitored sample had an average level of reported receivables of CZK 652 thousand, satisfaction of their creditors reached 58.8%. The main changes in the institute of personal bankruptcy are then evaluated in the closing discussion, and the impacts of these changes for households are assessed. The development of legislation in the Czech Republic and practice are shifting towards broader usage of personal bankruptcy, especially insofar as it can now also be used by entrepreneurs. Furthermore, the amendment of the Insolvency Act has enabled married couples to apply for joint debt relief, which has improved the position of the marriage partner with lower income and who would not get permission for debt relief on his/her own (mostly women are at issue). In current practice, the condition of adequate income is also solved by the fact that another person (usually a family member) undertakes to donate a certain monthly sum throughout the duration of the debt relief. Personal bankruptcy can thus be completed also by individuals to whom it would previously have been denied by the court.

Keywords: debtor, households, insolvency act, over-indebtedness, personal bankruptcy

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3 Mixotrophic Growth as a Tool for Increasing Polyhydroxyalkanoates (PHA) Production in Cyanobacteria

Authors: Zuzana Sedrlova, Eva Slaninova, Ines Fritz, Christina Daffert, Stanislav Obruca

Abstract:

Cyanobacteria are ecologically extremely important phototrophic gram-negative bacteria capable of oxygenic photosynthesis. They synthesize many interesting metabolites such as glycogen, carotenoids, but the most interesting metabolites are polyhydroxyalkanoates (PHA). The main advantage of cyanobacteria is the fact they do not require costly organic substrate and, oppositely, cyanobacteria can fix CO₂. PHA serves primarily as a carbon and energy source and occurs in the form of intracellular granules in bacterial cells. It is possible, PHA helps cyanobacteria to survive stress conditions since increased PHA synthesis was observed during cultivation in stress conditions. PHA is microbial biopolymers that are biodegradable with similar properties as petrochemical synthetic plastics. Production of PHA by heterotrophic bacteria is expensive; for price reduction waste materials as input, materials are used. Positively, cyanobacteria principally do not require organic carbon substrate since they are capable of CO₂ fixation. In this work, we demonstrated that stress conditions lead to the highest obtained yields of PHA in cyanobacterial cultures. Two cyanobacterial cultures from genera Synechocystis were used in this work. Cultivations were performed either in Erlenmayer flask or in tube multicultivator. Multiple stressors were applied on cyanobacterial cultures, and stressors include PHA precursors. PHA precursors are chemical substances and some of them do not occur naturally in the environment. Cultivation with the same PHA precursors in the same concentration led to a 1,6x higher amount of PHA when a multicultivator was used. The highest amount of PHA reached 25 % of PHA in dry cyanobacterial biomass. Both strains are capable of co-polymer synthesis in the presence of their structural precursor. The composition of co-polymer differs in Synechocystis sp. PCC 6803 and Synechocystis salina CCALA 192. Synechocystis sp. PCC 6803 cultivated with γ-butyrolakton accumulated co-polymer of 3-hydroxybutyrate (3HB) and 4-hydroxybutyrate (4HB) the composition of the copolymer was 56 % of 4HB and 44 % of 3HB. The total amount of PHA, as well as yield of biomass, was lower than in control due to the toxic properties of γ-butyrolakton. Funding: This study was partly funded by the project GA19- 19-29651L of the Czech Science Foundation (GACR) and partly funded by the Austrian Science Fund (FWF), a project I 4082-B25. This work was supported by Brno, Ph.D. Talent – Funded by the Brno City Municipality.

Keywords: co-polymer, cyanobacteria, PHA, synechocystis

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2 Detection, Analysis and Determination of the Origin of Copy Number Variants (CNVs) in Intellectual Disability/Developmental Delay (ID/DD) Patients and Autistic Spectrum Disorders (ASD) Patients by Molecular and Cytogenetic Methods

Authors: Pavlina Capkova, Josef Srovnal, Vera Becvarova, Marie Trkova, Zuzana Capkova, Andrea Stefekova, Vaclava Curtisova, Alena Santava, Sarka Vejvalkova, Katerina Adamova, Radek Vodicka

Abstract:

ASDs are heterogeneous and complex developmental diseases with a significant genetic background. Recurrent CNVs are known to be a frequent cause of ASD. These CNVs can have, however, a variable expressivity which results in a spectrum of phenotypes from asymptomatic to ID/DD/ASD. ASD is associated with ID in ~75% individuals. Various platforms are used to detect pathogenic mutations in the genome of these patients. The performed study is focused on a determination of the frequency of pathogenic mutations in a group of ASD patients and a group of ID/DD patients using various strategies along with a comparison of their detection rate. The possible role of the origin of these mutations in aetiology of ASD was assessed. The study included 35 individuals with ASD and 68 individuals with ID/DD (64 males and 39 females in total), who underwent rigorous genetic, neurological and psychological examinations. Screening for pathogenic mutations involved karyotyping, screening for FMR1 mutations and for metabolic disorders, a targeted MLPA test with probe mixes Telomeres 3 and 5, Microdeletion 1 and 2, Autism 1, MRX and a chromosomal microarray analysis (CMA) (Illumina or Affymetrix). Chromosomal aberrations were revealed in 7 (1 in the ASD group) individuals by karyotyping. FMR1 mutations were discovered in 3 (1 in the ASD group) individuals. The detection rate of pathogenic mutations in ASD patients with a normal karyotype was 15.15% by MLPA and CMA. The frequencies of the pathogenic mutations were 25.0% by MLPA and 35.0% by CMA in ID/DD patients with a normal karyotype. CNVs inherited from asymptomatic parents were more abundant than de novo changes in ASD patients (11.43% vs. 5.71%) in contrast to the ID/DD group where de novo mutations prevailed over inherited ones (26.47% vs. 16.18%). ASD patients shared more frequently their mutations with their fathers than patients from ID/DD group (8.57% vs. 1.47%). Maternally inherited mutations predominated in the ID/DD group in comparison with the ASD group (14.7% vs. 2.86 %). CNVs of an unknown significance were found in 10 patients by CMA and in 3 patients by MLPA. Although the detection rate is the highest when using CMA, recurrent CNVs can be easily detected by MLPA. CMA proved to be more efficient in the ID/DD group where a larger spectrum of rare pathogenic CNVs was revealed. This study determined that maternally inherited highly penetrant mutations and de novo mutations more often resulted in ID/DD without ASD in patients. The paternally inherited mutations could be, however, a source of the greater variability in the genome of the ASD patients and contribute to the polygenic character of the inheritance of ASD. As the number of the subjects in the group is limited, a larger cohort is needed to confirm this conclusion. Inherited CNVs have a role in aetiology of ASD possibly in combination with additional genetic factors - the mutations elsewhere in the genome. The identification of these interactions constitutes a challenge for the future. Supported by MH CZ – DRO (FNOl, 00098892), IGA UP LF_2016_010, TACR TE02000058 and NPU LO1304.

Keywords: autistic spectrum disorders, copy number variant, chromosomal microarray, intellectual disability, karyotyping, MLPA, multiplex ligation-dependent probe amplification

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1 Delicate Balance between Cardiac Stress and Protection: Role of Mitochondrial Proteins

Authors: Zuzana Tatarkova, Ivana Pilchova, Michal Cibulka, Martin Kolisek, Peter Racay, Peter Kaplan

Abstract:

Introduction: Normal functioning of mitochondria is crucial for cardiac performance. Mitochondria undergo mitophagy and biogenesis, and mitochondrial proteins are subject to extensive post-translational modifications. The state of mitochondrial homeostasis reflects overall cellular fitness and longevity. Perturbed mitochondria produce less ATP, release greater amounts of reactive molecules, and are more prone to apoptosis. Therefore mitochondrial turnover is an integral aspect of quality control in which dysfunctional mitochondria are selectively eliminated through mitophagy. Currently, the progressive deterioration of physiological functions is seen as accumulation of modified/damaged proteins with limiting regenerative ability and disturbance of such affected protein-protein communication throughout aging in myocardial cells. Methodologies: For our study was used immunohistochemistry, biochemical methods: spectrophotometry, western blotting, immunodetection as well as more sophisticated 2D electrophoresis and mass spectrometry for evaluation protein-protein interactions and specific post-translational modification. Results and Discussion: Mitochondrial stress response to reactive species was evaluated as electron transport chain (ETC) complexes, redox-active molecules, and their possible communication. Protein-protein interactions revealed a strong linkage between age and ETC protein subunits. Redox state was strongly affected in senescent mitochondria with shift in favor of more pro-oxidizing condition within cardiomyocytes. Acute myocardial ischemia and ischemia-reperfusion (IR) injury affected ETC complexes I, II and IV with no change in complex III. Ischemia induced decrease in total antioxidant capacity, MnSOD, GSH and catalase activity with recovery in some extent during reperfusion. While MnSOD protein content was higher in IR group, activity returned to 95% of control. Nitric oxide is one of the biological molecules that can out compete MnSOD for superoxide and produce peroxynitrite. This process is faster than dismutation and led to the 10-fold higher production of nitrotyrosine after IR injury in adult with higher protection in senescent ones. 2D protein profiling revealed 140 mitochondrial proteins, 12 of them with significant changes after IR injury and 36 individual nitrotyrosine-modified proteins further identified by mass spectrometry. Linking these two groups, 5 proteins were altered after IR as well as nitrated, but only one showed massive nitration per lowering content of protein after IR injury in adult. Conclusions: Senescent cells have greater proportion of protein content, which might be modulated by several post-translational modifications. If these protein modifications are connected to functional consequences and protein-protein interactions are revealed, link may lead to the solution. Assume all together, dysfunctional proteostasis can play a causative role and restoration of protein homeostasis machinery is protective against aging and possibly age-related disorders. This work was supported by the project VEGA 1/0018/18 and by project 'Competence Center for Research and Development in the field of Diagnostics and Therapy of Oncological diseases', ITMS: 26220220153, co-financed from EU sources.

Keywords: aging heart, mitochondria, proteomics, redox state

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