Genetic and molecular basis of neurodegenerative and neuropsychiatric diseases

Metody sekvenování DNA druhé (NGS) a třetí generace (TGS) zásadním způsobem ovlivnily strategii identifikace genetických příčin nemocí. Především použití exomového sekvenování zvýšilo účinnost identifikace kauzálních variant ve studovaných souborech pacientů z jednotek na desítky procent. Vzácné neurodegenerativní nemoci jsou unikátní svou klinickou i genetickou heterogenitou a širokou diferenciální diagnostikou. NGS a TGS technologie sehrály klíčovou roli v porozumění patomechanismu vzácných neurodegenerativních nemocí. Použití těchto technologií cestou výzkumu je u mnoha pacientů jedinou možností pro stanovení správné diagnózy, poskytnutí genetického poradenství a pro indikaci adekvátní léčby. Tato dizertační práce se zaměřuje na studium molekulární příčiny vybraných vzácných neurodegenerativních nemocí - adultní neuronální ceroidní lipofuscinózy (ANCL), spinální svalové atrofie (SMA) a neuronální intranukleární inkluzní nemoci (NIID). Studium molekulární příčiny těchto klinických jednotek s využitím moderních metod sekvenování a analýzy DNA vedlo k identifikaci kauzálních variant v souboru ANCL pacientů, vytvoření metodického postupu pro genetické testování SMA pacientů s negativním SMN1 nálezem a navržení metody pro vyšetření expanze tandemových repetic u NIID. Klíčová slova: adultní neuronální...Next-generation (NGS) and third-generation (TGS) sequencing methods have played a key role in strategies of disease genes identification. Especially the exome sequencing increased the efficiency of causal variants identification up to tens of percent in study cohorts. Rare neurodegenerative diseases are clinically and genetically heterogeneous and show a broad differential diagnostics. NGS and TGS technologies have been crucial in our understanding of the pathomechanism of rare neurodegenerative diseases. NGS and TGS, used by research laboratories, have been essential for many patients to determine a correct diagnosis, provide genetic counselling and reach an adequate treatment. This thesis focuses on molecular mechanisms of selected rare neurodegenerative diseases, namely adult neuronal ceroid lipofuscinosis (ANCL), spinal muscular atrophy (SMA) and neuronal intranuclear inclusion disease (NIID). Modern DNA sequencing methods led to identification of causal lesions in ANCL suspect patients. We provide a concept of genetic testing for SMN1 negative SMA patients and present a method for validation of tandem repeat expansion in NIID. Key words: adult neuronal ceroid lipofuscinosis, spinal muscular atrophy, neuronal intranuclear inclusion disease, next-generation sequencing methods, DNAJC5Institute of Inherited Metabolic Disorders First Faculty of Medicine Charles University in PragueÚstav dědičných metabolických poruch 1.LF a VFN v Praze1. lékařská fakultaFirst Faculty of Medicin

Evaluation of the impact of technical and technological adjustments to the water treatment plant on the quality of treated water

Předmětem této bakalářské práce je vyhodnocení postupné technické a technologické změny jímání a úpravy vody na konkrétní úpravně vody. Pro stanovení jsem si vybrala úpravnu vody v Ivančicích. První část této bakalářské práce vysvětluje základní pojmy týkající se problematiky tématu. Prvně je tedy seznámení s technologickými procesy úpravny vody, dále rozdělení úpraven vod, popis jakosti vod a technologií, které se na úpravně vody v Ivančicích nacházejí. V následujících dvou kapitolách se práce zaměřuje na popis úpravny vody před intenzifikací a po její intenzifikaci. Pozornost je věnována zejména odběru surové vody, jakosti surové i upravené vody a technologii úpravny vody. V poslední části této práce je provedeno srovnání původní a rekonstruované úpravny vody v několika hlediscích a následné doporučení pro zlepšení chodu rekonstruované úpravny vody. Důležité podklady pro hodnocení byly získány hlavně osobní prohlídkou úpravny vody. Z těchto a dalších podkladů se následně provedlo celkové hodnocení.The subject of this bachelor thesis is the evaluation of sequential technical and technological changes of water collection and modification at a specific water treatment facility – the water treatment facility in Ivančice. The first part of the thesis explains the basic terminology such as technological processes of the water treatment plant, types of water treatment facilities, the range of different types of water based on its quality and technologies that can be found at the water treatment plant in Ivančice. The next two chapters describe the water treatment process before and after intensification. The primary focus is on raw water collection, the quality of both raw and treated water, and the water treatment technology. The last part of the paper offers multiple comparisons of the original and renovated facility and recommendations for improving the operations of the renovated water treatment facility. The final evaluation was carried out based on the information collected mainly by visiting the facility in person, and other sources.

Heterozygous Loss-of-Function SEC61A1 Mutations Cause Autosomal-Dominant Tubulo-Interstitial and Glomerulocystic Kidney Disease with Anemia

Autosomal-dominant tubulo-interstitial kidney disease (ADTKD) encompasses a group of disorders characterized by renal tubular and interstitial abnormalities, leading to slow progressive loss of kidney function requiring dialysis and kidney transplantation. Mutations in UMOD, MUC1, and REN are responsible for many, but not all, cases of ADTKD. We report on two families with ADTKD and congenital anemia accompanied by either intrauterine growth retardation or neutropenia. Ultrasound and kidney biopsy revealed small dysplastic kidneys with cysts and tubular atrophy with secondary glomerular sclerosis, respectively. Exclusion of known ADTKD genes coupled with linkage analysis, whole-exome sequencing, and targeted re-sequencing identified heterozygous missense variants in SEC61A1—c.553A>G (p.Thr185Ala) and c.200T>G (p.Val67Gly)—both affecting functionally important and conserved residues in SEC61. Both transiently expressed SEC6A1A variants are delocalized to the Golgi, a finding confirmed in a renal biopsy from an affected individual. Suppression or CRISPR-mediated deletions of sec61al2 in zebrafish embryos induced convolution defects of the pronephric tubules but not the pronephric ducts, consistent with the tubular atrophy observed in the affected individuals. Human mRNA encoding either of the two pathogenic alleles failed to rescue this phenotype as opposed to a complete rescue by human wild-type mRNA. Taken together, these findings provide a mechanism by which mutations in SEC61A1 lead to an autosomal-dominant syndromic form of progressive chronic kidney disease. We highlight protein translocation defects across the endoplasmic reticulum membrane, the principal role of the SEC61 complex, as a contributory pathogenic mechanism for ADTKD

Evaluation of the impact of technical and technological adjustments to the water treatment plant on the quality of treated water

The subject of this bachelor thesis is the evaluation of sequential technical and technological changes of water collection and modification at a specific water treatment facility – the water treatment facility in Ivančice. The first part of the thesis explains the basic terminology such as technological processes of the water treatment plant, types of water treatment facilities, the range of different types of water based on its quality and technologies that can be found at the water treatment plant in Ivančice. The next two chapters describe the water treatment process before and after intensification. The primary focus is on raw water collection, the quality of both raw and treated water, and the water treatment technology. The last part of the paper offers multiple comparisons of the original and renovated facility and recommendations for improving the operations of the renovated water treatment facility. The final evaluation was carried out based on the information collected mainly by visiting the facility in person, and other sources

Genetic and molecular basis of neurodegenerative and neuropsychiatric diseases

Next-generation (NGS) and third-generation (TGS) sequencing methods have played a key role in strategies of disease genes identification. Especially the exome sequencing increased the efficiency of causal variants identification up to tens of percent in study cohorts. Rare neurodegenerative diseases are clinically and genetically heterogeneous and show a broad differential diagnostics. NGS and TGS technologies have been crucial in our understanding of the pathomechanism of rare neurodegenerative diseases. NGS and TGS, used by research laboratories, have been essential for many patients to determine a correct diagnosis, provide genetic counselling and reach an adequate treatment. This thesis focuses on molecular mechanisms of selected rare neurodegenerative diseases, namely adult neuronal ceroid lipofuscinosis (ANCL), spinal muscular atrophy (SMA) and neuronal intranuclear inclusion disease (NIID). Modern DNA sequencing methods led to identification of causal lesions in ANCL suspect patients. We provide a concept of genetic testing for SMN1 negative SMA patients and present a method for validation of tandem repeat expansion in NIID. Key words: adult neuronal ceroid lipofuscinosis, spinal muscular atrophy, neuronal intranuclear inclusion disease, next-generation sequencing methods, DNAJC

Pilot study on the risk assessment of selected water supply

Diplomová práce je zaměřena na posouzení rizik vodovodu pro veřejnou potřebu. Analýza rizik je řešena v souladu s metodikou vyvinutou VODÁRENSKOU AKCIOVOU SPOLEČNOSTÍ, a.s. a metodikou WaterRisk, kterou vytvořil Ústav vodního hospodářství obcí Fakulty stavební VUT v Brně. Práce vysvětluje základní pojmy týkající se dané problematiky tématu. Její připravenost k přijetí v České Republice a její podstatu vzhledem k legislativě. Dále se práce zaměřuje na popis konkrétního vodovodu. V následujících dvou kapitolách jsou posuzovány rizika podle vybraných metodik.This diploma thesis focuses on risk evaluation of water line for public use. The risk analysis is done in compliance with the methodology created by VODÁRENSKÁ AKCIOVÁ SPOLEČNOST, a. s. and by the method WaterRisk, created by Institute of Municipal Water management, Brno University of Technology, Faculty of Civil Engineering. The thesis clarifies the key points of the topic in question. Its readiness for passing in the Czech Republic and its core in relation to legislature. Then the thesis also includes a description of a specific water line. In the following two chapters the risks are studied according to the methodologies chosen.