Finding Conditions for the Separation and Identification of Protein Complexes by Using "Blue Native PAGE" and "Photo-induced Cross-Linking of Unmodified Proteins"

Department of Cell BiologyKatedra buněčné biologieFaculty of SciencePřírodovědecká fakult

Cytokinová exprese v chemicky indukované senescenci

AbstractJanderova-Rossmeislova L., Novakova Z., Vlasakova J., Philimonenko V., Hozak P., Hodny Z., 2007. PML protein association with specific nucleolar structures differs in normal, tumor and senescent human cells. J. Struct. Biol. 159, 56-70. Cellular senescence, widely recognized as a potent suppressor of tumorigenesis, represents response to cellular stress and DNA damage which results in irreversible cell growth arrest. Growing evidence signalizes crucial role of cytokine production in senescence phenomenon. Recently, many research groups have efforted to define exact character of senescence-associated secretory phenotype and its function in senescence development and maintentance. Factors secreted by senescent cells, mainly of proiflammatory character, were found to have pronounced effects on their environment as well as on its own producer. These observations were obtained preferentially on models of replicative senescence and oncogene-induced senescence. Vlasakova J, Novakova Z, Rossmeislova L, Kahle M, Hozak P, Hodny Z. 2007. Histone deacetylase inhibitors suppress IFNalpha-induced up-regulation of promyelocytic leukemia protein. Blood 109: 1373-80 Novakova Z, Man P, Novak P, Hozak P, Hodny Z. 2006. Separation of nuclear protein complexes by blue native polyacrylamide gel electrophoresis....Souhrn Buněčná senescence, kerá dokáže potlačit nádorové bujení, představuje odpověď buněk na stres a poškození DNA a způsobuje zástavu proliferace. Na tomto ději se zřejmě podílejí i produkované cytokiny. V současné době se mnoho vědeckých skupin snaží stanovit povahu a funkci sekrečního fenotypu spojeného se senescencí. Bylo zjištěno, že prozánětlivé látky sekretované senescentními buňkami ovlivňují okolní buňky i své vlastní producenty. Tyto poznatky byly získány především na modelech replikativní a onkogenem indukované senescence. Úkol této práce spočíval v definování cytokinové produkce senescentními buňkami indukovanými genotoxickými látkami bromodeoxyuridinem a distamycinem A. V tomto modelu jsme zjistili trvale aktivovanou interferonovou signalizaci spojenou se zvýšením interferonových cílových genů zahrnujících PML, STAT1 a IRF1. Při testování více než 160 cytokinů a jejich receptorů jsme nalezli zvýšenou hladinu u více než dvaceti cytokinů zahrnující prozánětlivé faktory IL-1, IL-6 and IL-8. Domníváme se, že cytokinová signalizace spojená se senescencí vyvolavou chemikáliemi představuje složitý a robustní děj, který je udržován pozitivními zpětnými vazbami. Charles University in Prague Faculty of Sciences PML (promyelocytární leukemický protein) a PML tělíska hrají roli v potlačování...Department of Cell BiologyKatedra buněčné biologieFaculty of SciencePřírodovědecká fakult

Optimization of izolation technique utilized for preparation of template material suitable for gene expression analysis of mesenchymal stromal cells from umbilical cord tissue

Ribonucleic acid (RNA) is the main component of the regulation of gene expression in the cell. As the development of techniques of molecular biology proceeds, RNA becomes an important tool for the gene expression analysis in research as well as diagnostic approaches. In contrast to deoxyribonucleic acid, RNA is very unstable and its isolation is tricky. RNA used for the gene expression analysis should fulfill several re foquirements focused especially on its purity. The isolation techniques should ensure effective separation of RNA from other cell components and chemicals used in the process. Currently, several methodologies are employed for the RNA isolation. The most common isolation is guanidine thiocyanate-phenol-chloroform combination called TRIzol or TRI Reagent and the extraction on solid phase so-called the column extraction. This study shows basic knowledge about macromolecule RNA and describes methodologies of RNA extraction from biological material. Spectrophotometry is very common technique used in laboratories. Thus the spectrophotometer is considered according to its speed, accuracy and simplicity of utilization. New improved devices reveal higher accuracy and lower requirements for the sample volume and make huge analyses much easier and more accurate. Getting knowledge about the principle and construction of the device we can improve utilization of the device and interpret the data correctly. In this study basic physical laws and principles concerning electromagnetic radiation and spectrophotometry are described, moreover, the basic model of spectrophotometer and critical parameters for nucleic acid quantification are shown. The aim of the study was to find the methodic approach for the isolation of unlimited amount of RNA of high quality suitable for high-throughput gene expression analysis of mesenchymal stromal cells. In the study several isolation techniques were compared to gain RNA suitable for reverse transcription reaction. These include isolation with guanidine thiocyanate-phenol-chloroform combination and column extraction. In total, yield and purity of RNA isolated by seven kits was compared in relation to the amount of cells used. The study involved comparison of two spectrophotometric devices NanoDrop ND-1000 and BioPhotometer Eppendorf. The accuracy of measurement of the concentration and the purity of RNA, speed, sample volume requirement and the weight of data acquired was compared between tested devices. A set of cell samples containing various number of mesenchymal stromal cells and commercially available RNA were used for testing purposes. The results of the study showed that in case of small cell number samples RNA isolated by column extraction is of higher quality than that isolated by TRI Reagent. From column-based kits the ZR RNA MicroPrep gave the best results the highest yield and purity of isolated RNA. From the spectrophotometric devices tested NanoDrop revealed smaller deviation of measurement, shorter time required for measurement, smaller sample volume requirement and higher weight of acquired data. In summary, ZR RNA MicroPrep and NanoDrop spectrophotometer were selected for preparation of RNA samples suitable for gene expression analysis

When God comes to know: The verbal root j-d-a as the predicate of God in Pentateuch and its comperhension in Rabbinic and Patristic literature

This Diploma thesis deals with two main issues. First, it explores the verbal root yadaʽ (know, learn) in the Pentateuch on places where God or the Lord is a grammatical subject, and second, how this verb has been interpreted by the rabbinic and patristic literature. The first part of the thesis focuses on general characteristics of the verb in contexts where it can reliably testify about God as the subject matter. The second part inquiries into exegesis of two particular cases of appearance of the verb yadaʽ in the so called intransitive form which occurs in the Pentateuch only twice, in Gen 18:21 and Ex 2:25. These two incidences are interconnected not only by the use of the verb yadaʽ but also by the complexity of explication and a number of other motives, especially the term of groaning. Both of the cases represent a groaning cry of the suffering people which is followed by God's response. While in Gen 18:21, the groaning comes from Sodom, in Ex 2:25, there are the sons of Israel crying from Egypt. However, God's knowledge does not precede these situations, it is a response to the resounding crying. The third part of my thesis analyses how the two given contexts are interpreted by rabbinic and patristic literature which can be regarded as sisterly traditions of interpretation of the first centuries of..

Cytokine expression in chemically-induced senescence

AbstractJanderova-Rossmeislova L., Novakova Z., Vlasakova J., Philimonenko V., Hozak P., Hodny Z., 2007. PML protein association with specific nucleolar structures differs in normal, tumor and senescent human cells. J. Struct. Biol. 159, 56-70. Cellular senescence, widely recognized as a potent suppressor of tumorigenesis, represents response to cellular stress and DNA damage which results in irreversible cell growth arrest. Growing evidence signalizes crucial role of cytokine production in senescence phenomenon. Recently, many research groups have efforted to define exact character of senescence-associated secretory phenotype and its function in senescence development and maintentance. Factors secreted by senescent cells, mainly of proiflammatory character, were found to have pronounced effects on their environment as well as on its own producer. These observations were obtained preferentially on models of replicative senescence and oncogene-induced senescence. Vlasakova J, Novakova Z, Rossmeislova L, Kahle M, Hozak P, Hodny Z. 2007. Histone deacetylase inhibitors suppress IFNalpha-induced up-regulation of promyelocytic leukemia protein. Blood 109: 1373-80 Novakova Z, Man P, Novak P, Hozak P, Hodny Z. 2006. Separation of nuclear protein complexes by blue native polyacrylamide gel electrophoresis...

When God comes to know: The verbal root j-d-a as the predicate of God in Pentateuch and its comperhension in Rabbinic and Patristic literature

This Diploma thesis deals with two main issues. First, it explores the verbal root yadaʽ (know, learn) in the Pentateuch on places where God or the Lord is a grammatical subject, and second, how this verb has been interpreted by the rabbinic and patristic literature. The first part of the thesis focuses on general characteristics of the verb in contexts where it can reliably testify about God as the subject matter. The second part inquiries into exegesis of two particular cases of appearance of the verb yadaʽ in the so called intransitive form which occurs in the Pentateuch only twice, in Gen 18:21 and Ex 2:25. These two incidences are interconnected not only by the use of the verb yadaʽ but also by the complexity of explication and a number of other motives, especially the term of groaning. Both of the cases represent a groaning cry of the suffering people which is followed by God's response. While in Gen 18:21, the groaning comes from Sodom, in Ex 2:25, there are the sons of Israel crying from Egypt. However, God's knowledge does not precede these situations, it is a response to the resounding crying. The third part of my thesis analyses how the two given contexts are interpreted by rabbinic and patristic literature which can be regarded as sisterly traditions of interpretation of the first centuries of..

When God comes to know: The verbal root j-d-a as the predicate of God in Pentateuch and its comperhension in Rabbinic and Patristic literature

Tato diplomová práce se zabývá jednak charakteristikou slovesného kořene j-d-a (vědět, poznat) v Pentateuchu na místech, kde Bůh či Hospodin je subjektem, a jednak tím, jak toto sloveso vykládá rabínská a patristická literatura. První část práce se věnuje obecné charakteristice tohoto slovesa v kontextech, kde může hodnotně vypovědět něco o Bohu či Hospodinu jako o svém subjektu. Druhá část práce se zabývá exegezemi dvou specifických výskytů slovesa j-d-a v tzv. bezpředmětné vazbě, která se vyskytuje v Pentateuchu pouze na dvou místech, v Gn 18,21 a Ex 2,25. Tyto dva výskyty spojuje vedle slovesa j-d-a i obtížnost jejich výkladu a řada dalších motivů, zejména termín úpění. V obou případech jde o úpěnlivé volání trpících, na které navazuje Boží reakce. V Gn 18,21 jde o úpění přicházející ze Sodomy, v Ex 2,25 naopak úpěnlivě volají synové Izraele z Egypta. Boží poznání však nepředchází těmto situacím, nýbrž je reakcí na ozývající se křik. Třetí část práce se zabývá tím, jakým způsobem uvedená dvě místa vykládá rabínská a patristická literatura, které se dají považovat za sesterské vykladačské tradice prvních století křesťanského letopočtu. Powered by TCPDF (www.tcpdf.org)This Diploma thesis deals with two main issues. First, it explores the verbal root yadaʽ (know, learn) in the Pentateuch on places where God or the Lord is a grammatical subject, and second, how this verb has been interpreted by the rabbinic and patristic literature. The first part of the thesis focuses on general characteristics of the verb in contexts where it can reliably testify about God as the subject matter. The second part inquiries into exegesis of two particular cases of appearance of the verb yadaʽ in the so called intransitive form which occurs in the Pentateuch only twice, in Gen 18:21 and Ex 2:25. These two incidences are interconnected not only by the use of the verb yadaʽ but also by the complexity of explication and a number of other motives, especially the term of groaning. Both of the cases represent a groaning cry of the suffering people which is followed by God's response. While in Gen 18:21, the groaning comes from Sodom, in Ex 2:25, there are the sons of Israel crying from Egypt. However, God's knowledge does not precede these situations, it is a response to the resounding crying. The third part of my thesis analyses how the two given contexts are interpreted by rabbinic and patristic literature which can be regarded as sisterly traditions of interpretation of the first centuries of...Katedra Starého zákonaDepartment of Old Testament StudiesEvangelická teologická fakultaProtestant Theological Facult

Cytokine expression in chemically-induced senescence

AbstractJanderova-Rossmeislova L., Novakova Z., Vlasakova J., Philimonenko V., Hozak P., Hodny Z., 2007. PML protein association with specific nucleolar structures differs in normal, tumor and senescent human cells. J. Struct. Biol. 159, 56-70. Cellular senescence, widely recognized as a potent suppressor of tumorigenesis, represents response to cellular stress and DNA damage which results in irreversible cell growth arrest. Growing evidence signalizes crucial role of cytokine production in senescence phenomenon. Recently, many research groups have efforted to define exact character of senescence-associated secretory phenotype and its function in senescence development and maintentance. Factors secreted by senescent cells, mainly of proiflammatory character, were found to have pronounced effects on their environment as well as on its own producer. These observations were obtained preferentially on models of replicative senescence and oncogene-induced senescence. Vlasakova J, Novakova Z, Rossmeislova L, Kahle M, Hozak P, Hodny Z. 2007. Histone deacetylase inhibitors suppress IFNalpha-induced up-regulation of promyelocytic leukemia protein. Blood 109: 1373-80 Novakova Z, Man P, Novak P, Hozak P, Hodny Z. 2006. Separation of nuclear protein complexes by blue native polyacrylamide gel electrophoresis...

The Influence of the Classroom Age Arrangement on Preschool Education

Přehledová studie si klade za cíl zmapovat výzkumy zaměřené na problematiku věkově heterogenních a homogenních tříd v mateřských školách. Této oblasti není v českém předškolním vzdělávání věnován žádný relevantní výzkum, přestože se s povinnou předškolní docházkou (od roku 2017) zvýšila četnost diskusí o vhodnosti heterogenního uspořádání tříd. Studie využívá 14 časopiseckých studií a dvou kapitol v monografiích ze šesti zemí, aby odpověděla na následující otázky: (1) Jaké cíle byly ve studiích sledovány? (2) Jaká podstatná z jištění a závěry z těchto studií vyplynuly? Shrnutí nálezů z vybraných studií naznačují směr, jakým by se mohl ubírat výzkum předškolního vzdělávání v českém prostředí mateřských škol.The main aim of this review study is to map research that was conducted on mixed-age and same-age preschool classrooms. In the Czech Republic, very little attention has been dedicated to these settings, although after the 2017 implementation of the compulsory preschool year, more attention and discussion is being paid to this research field. A total of 14 studies and 2 chapters from 6 countries were selected and analyzed to answer following questions: (1) What aims are pursued in these studies? and (2) What are the main results of the analyzed studies? A summary of the research findings indicates the orientation of possible preschool school level research in the Czech Republic