Helium permeation through mixed matrix membranes based on polyimides and silicalite-1

Mixed matrix membranes based on modified polyimides and silicalite-1 were prepared and studied. The novel preparation approach consists in improvement of the interfacial adhesion by employment of the coupling agent 3-aminopropyltriethoxysilane in such a way which leads to forming of chemical bonds between polyimide and silicalite-1. Firstly polyimide chains were endcapped by this agent which subsequently enabled their reaction with groups naturally present on silicalite-1 surface. Membranes with silicalite-1 content up to 60 wt. % were prepared and characterized by SEM, light microscopy and permeation of gases, prevailingly of He. The accessibility of sililicalite-1 pores prior to embedding and after embedding into polymeric matrices was studied by an iodine indicator technique. The permeability of membranes for gases was measured using a semi-open permeation apparatus with a small volume. Helium permeability depended on filler content and increased monotonously with the increasing content of filler. There were, however, significant deviations of the permeability dependences on filler content from shapes predicted by Bruggeman’s model. The qualitative explanation of the data disagreement with the model was proposed based on a stratification of mixed matrix membranes found by SEM

Pathway-specific effects of ADSL deficiency on neurodevelopment

Adenylosuccinate lyase (ADSL) functions in de novo purine synthesis (DNPS) and the purine nucleotide cycle. ADSL deficiency (ADSLD) causes numerous neurodevelopmental pathologies, including microcephaly and autism spectrum disorder. ADSLD patients have normal serum purine nucleotide levels but exhibit accumulation of dephosphorylated ADSL substrates, S-Ado, and SAICAr, the latter being implicated in neurotoxic effects through unknown mechanisms. We examined the phenotypic effects of ADSL depletion in human cells and their relation to phenotypic outcomes. Using specific interventions to compensate for reduced purine levels or modulate SAICAr accumulation, we found that diminished AMP levels resulted in increased DNA damage signaling and cell cycle delays, while primary ciliogenesis was impaired specifically by loss of ADSL or administration of SAICAr. ADSL-deficient chicken and zebrafish embryos displayed impaired neurogenesis and microcephaly. Neuroprogenitor attrition in zebrafish embryos was rescued by pharmacological inhibition of DNPS, but not increased nucleotide concentration. Zebrafish also displayed phenotypes commonly linked to ciliopathies. Our results suggest that both reduced purine levels and impaired DNPS contribute to neurodevelopmental pathology in ADSLD and that defective ciliogenesis may influence the ADSLD phenotypic spectrum.ID was funded by the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement no. 754510, THS, JL, and SP were funded by the Ministry of Science, Innovation and Universities (MCIU; PGC2018-095616-B-I00 to THS, PGC2018-099562-B-I00 to JL, and BFU2017-83562-P to SP), the 2017 SGR 1089 (AGAUR), FEDER, the Centres of Excellence Severo Ochoa award, and the CERCA Programme. THS was supported by the NIH Intramural Research Program, National Cancer Institute, Center for Cancer Research. MP was funded by grants from the Deutsche Forschungsgemeinschaft (DFG PH144/4-1 and PH144/6-1). MZ, OS, and VS were supported by Charles University, program PROGRES Q26/LF1. We would like to thank Biocev, First Faculty of Medicine, Charles University, for the opportunity to use their department’s equipment

A pathogenity of adenylosuccinate lyase deficiency

Adenylosuccinate lyase (ADSL) is an enzyme acting in two pathways of purine nucleotide metabolism. Mutations in ADSL gene compromising the enzyme activity lead to an inherited metabolic disease with severe neurological involvement - ADSL deficiency. Three distinct clinical phenotypes can be distinguished based on onset and severity of symptoms. The pathogenic mechanisms leading to the development of symptoms and underlying the phenotypic heterogeneity are unclear. The main pathogenic effect is attributed to the toxic effects of accumulating succinylpurines (SAdo, SAICAr). Their concentrations in cerebrospinal fluid, particularly diverse SAdo/SAICAr ratio, do correspond with the phenotypic groups. It is hypothesized that it may result from a mutation specific and thus structural related non-parallel loss (or gain) of enzyme activity towards one of its substrates. The main goal of the thesis is to seek for biochemical and structural basis of the diverse SAdo/SAICAr ratios and thus explain a pathogenetic mechanism of ADSL deficiency

The CRISPR-Cas9 crADSL HeLa Transcriptome: A First Step in Establishing a Model for ADSL Deficiency and SAICAR Accumulation

Adenylosuccinate lyase (ADSL) catalyzes two steps in de novo purine synthesis (DNPS). Mutations in ADSL can result in inborn errors of metabolism characterized by developmental delay and disorder phenotypes, with no effective treatment options. Recently, SAICAR, a metabolic substrate of ADSL, has been found to have alternative roles in the cell, complicating the role of ADSL. crADSL, a CRISPR KO of ADSL in HeLa cells, was constructed to investigate DNPS and ADSL in a human cell line. Here we employ this cell line in an RNA-seq analysis to initially investigate the effect of DNPS and ADSL deficiency on the transcriptome as a first step in establishing a cellular model of ADSL deficiency. We report transcriptome changes in genes relevant to development, vascular development, muscle, and cancer biology, which provide interesting avenues for future research

Improved diagnostics of purine and pyrimidine metabolism disorders using LC-MS/MS and its clinical application

Objectives: To develop a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify 41 different purine and pyrimidine (PuPy) metabolites in human urine to allow detection of most known disorders in this metabolic pathway and to determine reference intervals. Methods: Urine samples were diluted with an aqueous buffer to minimize ion suppression. For detection and quantification, liquid chromatography was combined with electrospray ionization, tandem mass spectrometry and multiple reaction monitoring. Transitions and instrument settings were established to quantify 41 analytes and nine stable-isotope-labeled internal standards (IS). Results: The established method is precise (intra-day CV: 1.4–6.3%; inter-day CV: 1.3–15.2%), accurate (95.2% external quality control results within ±2 SD and 99.0% within ±3 SD; analyte recoveries: 61–121%), sensitive and has a broad dynamic range to quantify normal and pathological metabolite concentrations within one run. All analytes except aminoimidazole ribonucleoside (AIr) are stable before, during and after sample preparation. Moreover, analytes are not affected by five cycles of freeze-thawing (variation: −5.6 to 7.4%), are stable in thymol (variation: −8.4 to 12.9%) and the lithogenic metabolites also in HCl conserved urine. Age-dependent reference intervals from 3,368 urine samples were determined and used to diagnose 11 new patients within 7 years (total performed tests: 4,206). Conclusions: The presented method and reference intervals enable the quantification of 41 metabolites and the potential diagnosis of up to 25 disorders of PuPy metabolism

Legislative Documents

Also, variously referred to as: House bills; House documents; House legislative documents; legislative documents; General Court documents

Helium permeation through mixed matrix membranes based on polyimides and silicalite-1

Mixed matrix membranes based on modified polyimides and silicalite-1 were prepared and studied. The novel preparation approach consists in improvement of the interfacial adhesion by employment of the coupling agent 3-aminopropyltriethoxysilane in such a way which leads to forming of chemical bonds between polyimide and silicalite-1. Firstly polyimide chains were endcapped by this agent which subsequently enabled their reaction with groups naturally present on silicalite-1 surface. Membranes with silicalite-1 content up to 60 wt. % were prepared and characterized by SEM, light microscopy and permeation of gases, prevailingly of He. The accessibility of sililicalite-1 pores prior to embedding and after embedding into polymeric matrices was studied by an iodine indicator technique. The permeability of membranes for gases was measured using a semi-open permeation apparatus with a small volume. Helium permeability depended on filler content and increased monotonously with the increasing content of filler. There were, however, significant deviations of the permeability dependences on filler content from shapes predicted by Bruggeman’s model. The qualitative explanation of the data disagreement with the model was proposed based on a stratification of mixed matrix membranes found by SEM

Study of the Accessibility of Zeolite Crystals in Polyimide Matrices. A Route to Coatings Exhibiting Selective Permeation

The accessibility of void space in MFI-type zeolite crystals, isolated and embedded in a polyimide (PI) matrix, was studied using optical microscopy coupled with an iodine indicator technique (IIT). IIT was used to estimate the adhesion between the PI matrix and the zeolitic phase, the accessibility of the zeolite void space for gas molecules as well as for characterizing the spatial distribution of embedded crystals in the composite. The channel system of the zeolitic phase in as-synthesized composites with the majority of crystals covered by a layer of PI is occupied by the solvent molecules used in composite synthesis. In untreated composites, the zeolitic phase is inaccessible to iodine sorption. Colouring patterns and colouring kinetics have been used to characterize the efficiency of the treatment for removing the PI covering layer from the crystal surface. The same techniques can be applied to characterize the treatment with regard to the desorption of solvents ( N- methyl-2-pyrrolidone, N,N -dimethylformamide, n-heptane) from the silicalite-1 channel system

Prague Dependency Treebank 3.0

PDT 3.0 is a new version of Prague Dependency Treebank. It contains a large amount of Czech texts with complex and interlinked morphological (2 million words), syntactic (1.5 MW) and semantic annotation (0.8 MW); in addition, certain properties of sentence information structure, multiword expressions, coreference, bridging relations and discourse relations are annotated at the semantic level