Analytical study of reduction of ammonium perrhenate using selected reduction agents

Tato práce je zaměřena na studium redukce rhenistanu amonného méně častými re- dukčními činidly. Jako méně častá redukční činidla byla zvolena kyselina askorbová a acetylhydrazin. Redukce byly sledovány pomocí metody kapilární zónové elektroforézy a UV-VIS spektrometrie. Měření proběhla v kyselém i bazickém prostředí. V případě metody kapilární zónové elektroforézy byl sledován úbytek plochy píku rhenistanu amonného po přidání redukčního činidla. U metody UV-VIS spektrometrie byla sledována míra absorpce rhenistanu amon- ného. Dále byla sledována možnost posunu rovnováhy reakce ve prospěch produktů redukce vlivem přídavku komplexotvorného činidla. Jako ligand byl použit pyrogallol (benzen- 1,2,3-triol). Jako redukční činidlo byl zvolen acetylhydrazin. Redukce byla sledována me- todou kapilární zónové elektroforézy. Identita vzniklého komplexu byla potvrzena s po- mocí ESI/MS. Klíčová slova rhenium, komplexní sloučeniny rhenia, kyselina askorbová, acetylhydrazin, kapilární zóno- vá elektroforéza, UV-VIS spektrometrie, hmotnostní spektrometrieThis thesis is focused on analysis of reduction of ammonium perrhenate using less frequent reducing agents. Ascorbic acid and acetylhydrazine were chosen as less common reducing agents. Reductions were observed leveraging capillary zone electrophoresis and UV/Vis spectroscopy. Measurements were carried out both in acidic and in basic environments. Reduction of peak area of ammonium perrhenate was observed using capillary zone electrophoresis after addition of reducing agent. Rate of absorption of ammonium perrhenate was monitored with UV/Vis spectroscopy. Furthermore, the possibility of shifting the equilibrium of the reaction to the benefit of the reduction products was monitored by the addition of the complexing agent. Py- rogallol (benzene-1,2,3-triol) was used as the ligand and acetylhydrazine was selected as the reducing agent. The reduction was monitored by capillary zone electrophoresis. The identity of the resulting complex was confirmed with ESI/MS. Keywords rhenium, rhenium complexes, ascorbic acid, acetylhydrazine, capillary zone electrophore- sis, UV/Vis spectroscopy, mass spectrometryKatedra analytické chemieDepartment of Analytical ChemistryPřírodovědecká fakultaFaculty of Scienc

Type of cell death and superinfection exclusion phenomenon in vaccinia virus-infected cells

Vaccinia virus (VACV) was formerly used in the eradication campaign against smallpox. VACV infection causes lysis of most cell types, including epithelial ones, which is equivalent to necrosis. However in our laboratory, we have previously detected activation of caspases during infection of HeLa G and BSC-40 epithelial cell lines with VACV, a typical sign of apoptosis. In this thesis, the type of cell death in HeLa G and BSC-40 cell lines infected with VACV strain WR was further studied and the activation and activity of caspases in the infected cells was characterized. In HeLa G cells infected with VACV strain WR, apoptosis was identified, as demonstrated by condensed nuclei, activity of caspase-3 and cleavage of death substrates. Additionally, activation and activity of caspase-2 and -4 was detected in infected HeLa G cells. In infected BSC-40 cells, neither apoptosis nor caspase activity were observed, but activation of caspase-2 and 4 was detected in these cells also. Finally, cleavage of procaspase-3 and -12 was detected in infected cells of both cell lines. Vaccination strains Praha and Dryvax induced apoptosis in both HeLa G and BSC-40 cells, as was demonstrated by the apoptotic morphology of nuclei and by the cleavage of PARP, substrate of the executioner caspases. Our results suggest that...Virus vakcinie (VACV) byl v minulosti využíván v eradikační kampani proti černým neštovicím. Infekce VACV je ve většině buněčných typů, včetně epiteliálních, lytická, což odpovídá nekrotické buněčné smrti. Při infekci epiteliálních linií HeLa G a BSC-40 VACV kmenem Western Reserve (WR) však byla v naší laboratoři již dříve pozorována aktivace kaspáz, která je typická pro apoptózu. V této práci byl blíže studován typ buněčné smrti buněk HeLa G a BSC-40 infikovaných VACV a také byla charakterizována aktivace a aktivita kaspáz u těchto infikovaných buněk. U buněk HeLa G infikovaných VACV kmene WR byla identifikována apoptóza na základě kondenzace jader, aktivity kaspázy 3 a štěpení substrátů smrti. V infikovaných buňkách HeLa G byla také detekována aktivace i aktivita kaspáz 2 a 4. U buněk BSC-40 infikovaných VACV kmenem WR apoptóza ani aktivita kaspáz pozorována nebyla, aktivace kaspáz 2 a 4 však zde byla detekována také. V infikovaných buňkách obou buněčných linií byla také štěpena prokaspáza 3 a 12. Vakcinační kmeny Dryvax a Praha indukovaly apoptózu v buňkách HeLa G i BSC-40, což bylo prokázáno apoptotickou morfologií jader a také detekcí štěpení substrátu exekutorových kaspáz PARP. Naše výsledky naznačují, že typ buněčné smrti infikovaných buněk může souviset s virulencí a imunogenitou různých...Department of Genetics and MicrobiologyKatedra genetiky a mikrobiologieFaculty of SciencePřírodovědecká fakult

Phytase-mediated enzymatic mineralization of chitosan-enriched hydrogels

Hydrogels mineralized with calcium phosphate (CaP) are increasingly popular bone regeneration biomaterials. Mineralization can be achieved by phosphatase enzyme incorporation and incubation in calcium glycerophosphate (CaGP). Gellan gum (GG) hydrogels containing the enzyme phytase and chitosan oligomer were mineralized in CaGP solution and characterized with human osteoblast-like MG63 cells and adipose tissue-derived stem cells (ADSC). Phytase induced CaP formation. Chitosan concentration determined mineralization extent and hydrogel mechanical reinforcement. Phytase-induced mineralization promoted MG63 adhesion and proliferation, especially in the presence of chitosan, and was non-toxic to MG63 cells (with and without chitosan). ADSC adhesion and proliferation were poor without mineralization. Chitosan did not affect ADSC osteogenic differentiation

Twist exome capture allows for lower average sequence coverage in clinical exome sequencing

Background Exome and genome sequencing are the predominant techniques in the diagnosis and research of genetic disorders. Sufficient, uniform and reproducible/consistent sequence coverage is a main determinant for the sensitivity to detect single-nucleotide (SNVs) and copy number variants (CNVs). Here we compared the ability to obtain comprehensive exome coverage for recent exome capture kits and genome sequencing techniques. Results We compared three different widely used enrichment kits (Agilent SureSelect Human All Exon V5, Agilent SureSelect Human All Exon V7 and Twist Bioscience) as well as short-read and long-read WGS. We show that the Twist exome capture significantly improves complete coverage and coverage uniformity across coding regions compared to other exome capture kits. Twist performance is comparable to that of both short- and long-read whole genome sequencing. Additionally, we show that even at a reduced average coverage of 70× there is only minimal loss in sensitivity for SNV and CNV detection. Conclusion We conclude that exome sequencing with Twist represents a significant improvement and could be performed at lower sequence coverage compared to other exome capture techniques

A Solve-RD ClinVar-based reanalysis of 1522 index cases from ERN-ITHACA reveals common pitfalls and misinterpretations in exome sequencing

Purpose Within the Solve-RD project (https://solve-rd.eu/), the European Reference Network for Intellectual disability, TeleHealth, Autism and Congenital Anomalies aimed to investigate whether a reanalysis of exomes from unsolved cases based on ClinVar annotations could establish additional diagnoses. We present the results of the “ClinVar low-hanging fruit” reanalysis, reasons for the failure of previous analyses, and lessons learned. Methods Data from the first 3576 exomes (1522 probands and 2054 relatives) collected from European Reference Network for Intellectual disability, TeleHealth, Autism and Congenital Anomalies was reanalyzed by the Solve-RD consortium by evaluating for the presence of single-nucleotide variant, and small insertions and deletions already reported as (likely) pathogenic in ClinVar. Variants were filtered according to frequency, genotype, and mode of inheritance and reinterpreted. Results We identified causal variants in 59 cases (3.9%), 50 of them also raised by other approaches and 9 leading to new diagnoses, highlighting interpretation challenges: variants in genes not known to be involved in human disease at the time of the first analysis, misleading genotypes, or variants undetected by local pipelines (variants in off-target regions, low quality filters, low allelic balance, or high frequency). Conclusion The “ClinVar low-hanging fruit” analysis represents an effective, fast, and easy approach to recover causal variants from exome sequencing data, herewith contributing to the reduction of the diagnostic deadlock

Assessment in Biology Education in Ceske Budejovice and Surroundings

VOMÁČKOVÁ J. 2015: Assessment in Biology Education in Ceske Budejovice and surroundings. Bachelor thesis. Faculty of Education, University of South Bohemia in Ceske Budejovice. 39 pp. Assessment is the key part to all educational and pedagogic activities. Based on this, assessment is nowadays often discussed not only in the academic sphere but also by teachers doing the practical job. This is especially viewed from the point of the effectiveness and adequacy of individual assessment methods in concrete subjects taught. The aim of this bachelor thesis was to monitor assessment methods currently used in Biology lessons. This has been done through questionnaires used with grammar school students and elementary school students attending the second level study in the Ceske Budejovice region. The results show assumptive assessment methods prevail with teachers mainly focusing on the students knowing definitions, facts and formulae. New activating methods are being introduced in Biology lessons, which makes the assessment of students' performance more difficult. Formative assessment might be the answer to this. In the part of the questionnaire where the students´ personal experience, preferences and opinions have been monitored, it has been found out students themselves consider this type of assessment more motivating and beneficial then the assumptive one

Komparace elektronických školních informačních systémů z hlediska využitelnosti pro MŠ a ZŠ

Tato práce se zabývá porovnáním nejčastěji používaných elektronických školních informačních systémů. Jedná se o systémy Bakaláři, dm Software, Škola Online a EduPage. Práce se také zaměřuje na možnosti jejich využití v podmínkách spojené základní a mateřské školy. Kromě rozsahu jednotlivých systémů, volitelných modulů, jejich ovládání a využitelnosti, se zaobírá i přehledností a uživatelskou přívětivostí, a to nejen z pohledu vedení školy a pedagogů, ale i z pohledu rodičů a žáků. Vzhledem k rozšíření smart technologií je část práce věnována i srovnání mobilních verzí jednotlivých produktů. Cílem práce je komparace výhod a nevýhod používání výše uvedených systémů, ale nikoli rozhodnutí, která varianta je nejlepší, neboť je třeba respektovat i individuální preference jednotlivých uživatelů

World Bank Development Agenda With Focus on Sub-Saharan Africa

World Bank Development Agenda With Focus on Sub-Saharan Africa Diploma thesis with subject World Bank Development Agenda With Focus on Sub-Ssaharan Africa is dividend into free main chapters. The first one deals with Sub-Saharan Africa as a development region. It briefly describes Africa's history and some of ecomomic and social characteristics, and also mentions main economic and social problems of the region. Second chapter concentrates on World Bank institution and the evolution of its development strategies since 1950s, in context with UN attitudes to development. Third charter looks into finance flows from World Bank to Subsaharan Africa since 1950s upto February 2012.Diplomová práce na téma Rozvojová agenda Světové banky se zaměřením na subsaharskou Afriku je členěna do tří hlavních kapitol. První kapitola se zabývá subsaharskou Afrikou jako rozvojovým regionem. Stručně popisuje historický vývoj a vybrané ekonomické a demografické charakteristiky subsaharské Afriky. Popisuje též hlavní problémy regionu, které dělí na ekonomické a sociální. Druhá kapitola nejprve představí instituce Světové banky a následně popíše vývoj jejích rozvojových strategií od 50. let, v kontextu přístupů OSN k rozvoji. Třetí kapitola se zabývá rozborem finančních toků ve formě půjček Světové banky do zemí subsaharské Afriky od 50. let do února 2012

Hiporehabilitation - Paradresage in person with multiple sclerosis

Klinika rehabilitačního lékařstvíDepartment of Rehabilitation MedicineThird Faculty of Medicine3. lékařská fakult

Homoxenous coccidia of caudate amphibia: diversity and developmental cycles

The study is focused on homoxenous coccidia of caudate amphibia. The aims include review of relevant literature information in the context of current knowledge on taxonomy, systematics and biology of coccidia in general and coprological examination of newts from two localities for a presence of coccidian oocysts in fecal samples