Claviceps paspali is used in the pharmaceutical industry for the production of ergot alkaloids. This fungus also biosynthesizes paspalitrems, indole diterpene (IDT) mycotoxins that cause significant economic losses in agriculture and represent safety concerns for ergot alkaloid manufacture. Here, we use Agrobacterium-mediated transformation to replace the idtP and the idtF genes in the IDT biosynthetic gene cluster of C. paspali with a selectable marker gene. We show that the ΔidtP knockout mutant produces paspaline, the first IDT intermediate of the pathway. The ΔidtF strain produces unprenylated IDTs such as paspalinine and paspaline. These experiments validate the function of idtP as the gene encoding the cytochrome P450 monooxygenase that oxidizes and demethylates paspaline to produce 13-desoxypaxilline, and that of idtF as the gene that encodes the α-prenyltransferase that prenylates paspalinine at the C20 or the C21 positions to yield paspalitrems A and C, respectively. In addition, we also show that axenic cultures of the wild type, the ΔidtP and the ΔidtF mutant C. paspali strains fail to produce an assembly of IDTs that are present in C. paspali-Paspalum spp. associations.
PURPOSE: To report a simultaneous occurrence of 2 rare corneal dystrophies. METHODS: A 30-year-old man with a family history of posterior polymorphous corneal dystrophy type 3 (PPCD3) was invited for ophthalmic examination. Sanger sequencing of the coding regions and intron/exon boundaries of disease-associated genes, ZEB1 and UBIAD1, was performed. RESULTS: The clinical findings suggested co-occurrence of PPCD3 and Schnyder corneal dystrophy in the proband. This dual diagnosis was supported by genetic findings. He was identified to carry a previously reported heterozygous nonsense mutation in ZEB1: c.2157C>G; p.(Tyr719*), and a novel heterozygous missense mutation in UBIAD1: c.569T>C; p.(Ile190Thr). The mother of the proband only carried c.2157C>G in ZEB1, and slit-lamp examination of her corneas showed endothelial lesions characteristic of PPCD3. The sister of the proband carried c.569T>C in UBIAD1 and had corneal crystal deposition in her anterior stroma consistent with the diagnosis of Schnyder corneal dystrophy. CONCLUSIONS: This case illustrates the coincidental occurrence of 2 rare and genetically distinct corneal dystrophies in a single patient. Furthermore, it highlights the need to perform comprehensive phenotyping in combination with appropriate genetic diagnostic testing to achieve an accurate diagnosis.
- MeSH
- dědičné dystrofie rohovky diagnóza genetika MeSH
- dospělí MeSH
- lidé MeSH
- mutace MeSH
- prenyltransferáza genetika MeSH
- transkripční faktor Zeb1 genetika MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
BACKGROUND: The purpose of this study was to identify the genetic cause and describe the clinical phenotype of Schnyder corneal dystrophy (SCD) in six unrelated probands. METHODS: We identified two white Czech, two white British and two South Asian families with a clinical diagnosis of SCD. Ophthalmic assessment included spectral domain optical coherence tomography (SD-OCT) of one individual with advanced disease, and SD-OCT and confocal microscopy of a child with early stages of disease. UBIAD1 coding exons were amplified and Sanger sequenced in each proband. A fasting serum lipid profile was measured in three probands. Paternity testing was performed in one family. RESULTS: A novel heterozygous c.527G>A; p.(Gly176Glu) mutation in UBIAD1 was identified in one Czech proband. In the second Czech proband, aged 6 years when first examined, a previously described de novo heterozygous c.289G>A; p.(Ala97Thr) mutation was found. Two probands of South Asian descent carried a known c.305G>A; p.(Asn102Ser) mutation in the heterozygous state. Previously reported heterozygous c.361C>T; p.(Leu121Phe) and c.308C>T; p.(Thr103Ile) mutations were found in two white British families. Although crystalline deposits were present in all probands the affected area was small in some individuals. Corneal arcus and stromal haze were the most prominent phenotypical feature in two probands. In the Czech probands, SD-OCT confirmed accumulation of reflective material in the anterior stroma. Crystalline deposits were visualized by confocal microscopy. Mild dyslipidemia was found in all three individuals tested. CONCLUSION: Although de novo occurrence of mutations in UBIAD1 is extremely rare, SCD should be considered in the differential diagnosis of bilateral corneal haze and/or crystal deposition, especially in children.
- MeSH
- dědičné dystrofie rohovky diagnóza genetika metabolismus MeSH
- dítě MeSH
- DNA genetika MeSH
- dospělí MeSH
- fenotyp MeSH
- konfokální mikroskopie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mutace * MeSH
- mutační analýza DNA MeSH
- optická koherentní tomografie metody MeSH
- prenyltransferáza genetika metabolismus MeSH
- rodokmen MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Cytokinins, a class of phytohormones, are adenine derivatives common to many different organisms. In plants, these play a crucial role as regulators of plant development and the reaction to abiotic and biotic stress. Key enzymes in the cytokinin synthesis and degradation in modern land plants are the isopentyl transferases and the cytokinin dehydrogenases, respectively. Their encoding genes have been probably introduced into the plant lineage during the primary endosymbiosis. To shed light on the evolution of these proteins, the genes homologous to plant adenylate isopentenyl transferase and cytokinin dehydrogenase were amplified from the genomic DNA of cyanobacterium Nostoc sp. PCC 7120 and expressed in Escherichia coli. The putative isopentenyl transferase was shown to be functional in a biochemical assay. In contrast, no enzymatic activity was detected for the putative cytokinin dehydrogenase, even though the principal domains necessary for its function are present. Several mutant variants, in which conserved amino acids in land plant cytokinin dehydrogenases had been restored, were inactive. A combination of experimental data with phylogenetic analysis indicates that adenylate-type isopentenyl transferases might have evolved several times independently. While the Nostoc genome contains a gene coding for protein with characteristics of cytokinin dehydrogenase, the organism is not able to break down cytokinins in the way shown for land plants.
- MeSH
- biologická evoluce MeSH
- cytokininy metabolismus MeSH
- Escherichia coli enzymologie růst a vývoj MeSH
- fylogeneze MeSH
- geneticky modifikované rostliny genetika růst a vývoj metabolismus MeSH
- molekulární sekvence - údaje MeSH
- mutace genetika MeSH
- mutageneze cílená MeSH
- Nostoc enzymologie genetika MeSH
- oxidoreduktasy genetika metabolismus MeSH
- prenyltransferáza genetika metabolismus MeSH
- regulace genové exprese enzymů MeSH
- rekombinantní proteiny metabolismus MeSH
- sekvence aminokyselin MeSH
- sekvenční homologie aminokyselin MeSH
- tabák enzymologie růst a vývoj MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Bisfosfonáty jsou nejvíce užívanou skupinou léků využívaných v terapii osteoporózy. Jedná se o syntetické látky, které se ukládají do kostní tkáně, kde prostřednictvím inhibičního působení na osteoklasty výrazně snižují kostní resorpci a zvyšují mineralizaci, objem i pevnost kosti. V klinických studiích byl prokázán jejich příznivý vliv na zvýšení denzity kostního minerálu (BMD) a na snížení rizika vertebrálních i nevertebrálních zlomenin. Dosud však nebylo jednoznačně stanoveno, jak dlouho mají být bisfosfonáty podávány.
- MeSH
- alendronát aplikace a dávkování farmakokinetika škodlivé účinky terapeutické užití MeSH
- bisfosfonáty * aplikace a dávkování farmakokinetika farmakologie klasifikace škodlivé účinky terapeutické užití MeSH
- časové faktory MeSH
- fraktury kostí * prevence a kontrola MeSH
- fraktury páteře prevence a kontrola MeSH
- hydroxymethylglutaryl-CoA-reduktasy chemie MeSH
- kostní denzita účinky léků MeSH
- lidé středního věku MeSH
- lidé MeSH
- osteoblasty účinky léků MeSH
- osteoporóza * farmakoterapie MeSH
- prenyltransferáza chemie MeSH
- remodelace kosti MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
