Mutation of Nogo-B Receptor, a Subunit of cis-Prenyltransferase, Causes a Congenital Disorder of Glycosylation

SummaryDolichol is an obligate carrier of glycans for N-linked protein glycosylation, O-mannosylation, and GPI anchor biosynthesis. cis-prenyltransferase (cis-PTase) is the first enzyme committed to the synthesis of dolichol. However, the proteins responsible for mammalian cis-PTase activity have not been delineated. Here we show that Nogo-B receptor (NgBR) is a subunit required for dolichol synthesis in yeast, mice, and man. Moreover, we describe a family with a congenital disorder of glycosylation caused by a loss of function mutation in the conserved C terminus of NgBR-R290H and show that fibroblasts isolated from patients exhibit reduced dolichol profiles and enhanced accumulation of free cholesterol identically to fibroblasts from mice lacking NgBR. Mutation of NgBR-R290H in man and orthologs in yeast proves the importance of this evolutionarily conserved residue for mammalian cis-PTase activity and function. Thus, these data provide a genetic basis for the essential role of NgBR in dolichol synthesis and protein glycosylation

Mutations causing medullary cystic kidney disease type 1 (MCKD1) lie in a large VNTR in MUC1 missed by massively parallel sequencing

While genetic lesions responsible for some Mendelian disorders can be rapidly discovered through massively parallel sequencing (MPS) of whole genomes or exomes, not all diseases readily yield to such efforts. We describe the illustrative case of the simple Mendelian disorder medullary cystic kidney disease type 1 (MCKD1), mapped more than a decade ago to a 2-Mb region on chromosome 1. Ultimately, only by cloning, capillary sequencing, and de novo assembly, we found that each of six MCKD1 families harbors an equivalent, but apparently independently arising, mutation in sequence dramatically underrepresented in MPS data: the insertion of a single C in one copy (but a different copy in each family) of the repeat unit comprising the extremely long (~1.5-5 kb), GC-rich (>80%), coding VNTR in the mucin 1 gene. The results provide a cautionary tale about the challenges in identifying genes responsible for Mendelian, let alone more complex, disorders through MPS

Mutations causing medullary cystic kidney disease type 1 (MCKD1) lie in a large VNTR in MUC1 missed by massively parallel sequencing

While genetic lesions responsible for some Mendelian disorders can be rapidly discovered through massively parallel sequencing (MPS) of whole genomes or exomes, not all diseases readily yield to such efforts. We describe the illustrative case of the simple Mendelian disorder medullary cystic kidney disease type 1 (MCKD1), mapped more than a decade ago to a 2-Mb region on chromosome 1. Ultimately, only by cloning, capillary sequencing, and de novo assembly, we found that each of six MCKD1 families harbors an equivalent, but apparently independently arising, mutation in sequence dramatically underrepresented in MPS data: the insertion of a single C in one copy (but a different copy in each family) of the repeat unit comprising the extremely long (~1.5-5 kb), GC-rich (>80%), coding VNTR in the mucin 1 gene. The results provide a cautionary tale about the challenges in identifying genes responsible for Mendelian, let alone more complex, disorders through MPS

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

Mutations causing medullary cystic kidney disease type 1 lie in a large VNTR in MUC1 missed by massively parallel sequencing

Although genetic lesions responsible for some mendelian disorders can be rapidly discovered through massively parallel sequencing of whole genomes or exomes, not all diseases readily yield to such efforts. We describe the illustrative case of the simple mendelian disorder medullary cystic kidney disease type 1 (MCKD1), mapped more than a decade ago to a 2-Mb region on chromosome 1. Ultimately, only by cloning, capillary sequencing and de novo assembly did we find that each of six families with MCKD1 harbors an equivalent but apparently independently arising mutation in sequence markedly under-represented in massively parallel sequencing data: the insertion of a single cytosine in one copy (but a different copy in each family) of the repeat unit comprising the extremely long (~1.5–5 kb), GC-rich (>80%) coding variable-number tandem repeat (VNTR) sequence in the MUC1 gene encoding mucin 1. These results provide a cautionary tale about the challenges in identifying the genes responsible for mendelian, let alone more complex, disorders through massively parallel sequencing.National Institutes of Health (U.S.) (Intramural Research Program)National Human Genome Research Institute (U.S.)Charles University (program UNCE 204011)Charles University (program PRVOUK-P24/LF1/3)Czech Republic. Ministry of Education, Youth, and Sports (grant NT13116-4/2012)Czech Republic. Ministry of Health (grant NT13116-4/2012)Czech Republic. Ministry of Health (grant LH12015)National Institutes of Health (U.S.) (Harvard Digestive Diseases Center, grant DK34854

Ovarian carcinomas - the influence of the Rho kinase family on cell migration

Author: Veronika Barešová Title: Ovarian carcinomas - the influence of Rho family kinases on cell migration Diploma thesis Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Ovarian carcinomas are the deadliest group of gynecological malignancies. This is due to a high rate of metastasis and acquired drug resistance. Metastasis is characterized by a phenotypic change of cells from epithelial to mesenchymal, resulting in the loss of cell-cell and cell-extracellular matrix contacts and subsequent cell movement. This process is called epithelial-mesenchymal transition and has been shown to also play a role in cell resistance to cisplatin. The movement of individual cells is mediated by the Rho GTPase family and can be divided into mesenchymal and amoeboid types. The amoeboid type is characterized by the Rho/ROCK signaling pathway, while mesenchymal movement is mediated by the Rac/Cdc42 pathways. In my thesis, we investigated the differences in expression of selected proteins in both signaling pathways in ovarian carcinoma cells. Initially, we focused on comparing the A2780 and A2780cis cell lines, which are resistant to cisplatin. We monitored markers of amoeboid movement, RhoA and phosphorylated myosin, and markers of mesenchymal movement, Wave-2,..

Expansion proposal for LunaCup Ltd.

Cílem bakalářské práce bylo navrhnout zahraniční expanzi společnosti LunaCup s.r.o. zaměřující se na výrobu a prodej menstruačních kalíšků. První část práce seznamuje čtenáře s dostupnou literatura a vysvětluje metodologii a práci s daty. V druhé části práce je provedena SWOT analýza společnosti i produktu. Na tyto analýzy navazuje scoring model, pomocí něhož je vybrán rakouský trh. K posouzení trhu je použita analýza PEST a Porterova analýza. Tyto analýzy jsou doplněny o komentář ke strategii expanze. V závěru práce jsou shrnuty klíčové poznatky a představena doporučení.The bachelor thesis aimed to propose a foreign expansion for LunaCup Ltd., which focuses on the production and sale of menstrual cups. The first part of the thesis introduces the reader to the available literature and explains the methodology and work with data. In the second part, the SWOT analysis of the company and the product is conducted. These analyzes are followed by the scoring model, which selects the Austrian market. To assess the market, PEST and Porter’s analysis is applied. They are completed by a commentary on the strategy of expansion. At the end of the thesis, key findings are summarized, and recommendations are given

Ovarian carcinomas - the influence of the Rho kinase family on cell migration

Author: Veronika Barešová Title: Ovarian carcinomas - the influence of Rho family kinases on cell migration Diploma thesis Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Ovarian carcinomas are the deadliest group of gynecological malignancies. This is due to a high rate of metastasis and acquired drug resistance. Metastasis is characterized by a phenotypic change of cells from epithelial to mesenchymal, resulting in the loss of cell-cell and cell-extracellular matrix contacts and subsequent cell movement. This process is called epithelial-mesenchymal transition and has been shown to also play a role in cell resistance to cisplatin. The movement of individual cells is mediated by the Rho GTPase family and can be divided into mesenchymal and amoeboid types. The amoeboid type is characterized by the Rho/ROCK signaling pathway, while mesenchymal movement is mediated by the Rac/Cdc42 pathways. In my thesis, we investigated the differences in expression of selected proteins in both signaling pathways in ovarian carcinoma cells. Initially, we focused on comparing the A2780 and A2780cis cell lines, which are resistant to cisplatin. We monitored markers of amoeboid movement, RhoA and phosphorylated myosin, and markers of mesenchymal movement, Wave-2,...Autor: Veronika Barešová Název: Ovariální karcinomy - vliv rodiny Rho kináz na migraci buněk Diplomová práce Univerzita Karlova, Farmaceutická fakulta v Hradci Králové Katedra biologických a lékařských věd Ovariální karcinomy jsou nejsmrtelnější skupinou gynekologických malignit. Je to dáno vysokou mírou metastázování a získané lékové rezistence. Metastázování je charakteristické změnou fenotypu buněk z epiteliálního na mezenchymální, čímž dochází ke ztrátě kontaktů buňka-buňka a buňka-extracelulární matrix a následnému pohybu buněk. Tento proces se nazývá epitelo-mezenchymální tranzice a bylo prokázáno, že také hraje roli v rezistenci buněk na cisplatinu. Pohyb jednotlivých buněk je zprostředkován rodinou Rho GTPáz a můžeme ho rozdělit na mezenchymální a améboidní typ. Pro améboidní je typická signální dráha Rho/ROCK, kdežto mezenchymální pohyb je zprostředkován drahami Rac/Cdc42. V mé diplomové práci jsme sledovali rozdíly v expresi vybraných proteinů obou signálních drah v buňkách ovariálních karcinomů. V první fázi jsme se zaměřili na porovnání buněčných linií A2780 a A2780cis, která je rezistentní na cisplatinu. Sledovali jsme markery améboidního pohybu RhoA a fosforylovaný myozin a markery pohybu mezenchymálního Wave-2, PAK a fosforylovaný PAK. Ve druhé fázi jsme sledovali vliv EGF a...Katedra biologických a lékařských vědDepartment of Biological and Medical SciencesFaculty of Pharmacy in Hradec KrálovéFarmaceutická fakulta v Hradci Králov

The nursing process about patients with cervical spinal injury on the spinal unit

Cílem bakalářské práce je seznámit s problematikou vzniku, léčbou a ošetřováním pacientů po úrazech krční páteře. V práci je vypracován dlouhodobý ošetřovatelský proces u konkrétního pacienta. Na závěr je popsána aplikace vypracovaného ošetřovatelského procesu v praxi.Dokončená práce s úspěšnou obhajobo

Specific aspects of 20th century artwork of educational programme in gallery

The thesis focuses on difference between theoretical and didactical aspects of work with artwork of so-called great styles (from the Romance art until art of 19. century), modern and postmodern artefact. It surveys the current thought of the early 21st century about art and visual culture. On the basis of this thought educational programs for gallery space come up. Within the framework of the thesis, two confrontational gallery educational programs have been proposed. At the end of the thesis, based on the comparison of these programs the specific aspects of 20th century artwork of educational programme in gallery have been formulated