Oriented immobilization of peptide-N-glycosidase F on a monolithic support for glycosylation analysis

In this paper, we report on a novel oriented peptide-N-glycosidase F (PNGase F) immobilization approach onto methacrylate based monolithic support for rapid, reproducible and efficient release of the N-linked carbohydrate moieties from glycoproteins. The glutathione-S-transferase-fusion PNGase F (PNGase F-GST) was expressed in Escherichia coli using regular vector technology. The monolithic pore surface was functionalized with glutathione via a succinimidyl-6-(iodoacetyl-amino)-hexanoate linker and the specific affinity of GST toward glutathione was utilized for the oriented coupling. This novel immobilization procedure was compared with reductive amination technique commonly used for non-oriented enzyme immobilization via primary amine functionalities. Both coupling approaches were compared using enzymatic treatment of several glycoproteins, such as ribonuclease B, fetuin and immunoglobulin G followed by MALDI/MS and CE-LIF analysis of the released glycans. Orientedly immobilized PNGase F via GST-glutathione coupling showed significantly higher activity, remained stable for several months, and allowed rapid release of various types of glycans (high-mannose, core fucosylated, sialylated, etc.) from glycoproteins. Complete protein deglycosylation was obtained as fast as in several seconds when using flow-through immobilized microreactors

Grandparenting, health, and well-being: a systematic literature review

Whether grandparenting is associated with improved health or well-being among older adults is a salient question in present-day aging societies. This systematic review compiles studies that consider the health or well-being outcomes of grandparenting, concerning (1) custodial grandparent families, where grandparents are raising grandchildren without parental presence; (2) three-generation households, where grandparents are living with adult children and grandchildren; and (3) non-coresiding grandparents, who are involved in the lives of their grandchildren. Review was based on literature searches conducted in September 2019 via Web of Science, PubMed, PsycINFO, and Ebsco. We screened 3868 abstracts across four databases, and by following the PRISMA guidelines, we identified 92 relevant articles (117 studies) that were published between 1978 and 2019. In 68% of cases, custodial grandparenting was associated with decreased health or well-being of grandparents. The few studies considering the health or well-being of grandparents living in three-generation households provided mixed findings (39% positive; 39% negative). Finally, in 69% of cases, involvement of non-coresiding grandparents was associated with improved grandparental outcomes; however, there was only limited support for the prediction that involved grandparenting being causally associated with grandparental health or well-being. Despite this, after different robustness checks (counting all nonsignificant results, taking into account the representativeness of the data and causal methodology), the main finding remains the same: the most negative results are found among custodial grandparents and three-generation households and most positive results among non-coresiding grandparents

Czech national e-infrastructure services for HEP

Czech e-infrastructure project e-INFRA CZ is the only project for ICT services defined on the Road Map of the Czech Republic for Large Infrastructures for Research, Experimental Development and Innovations. It was created by 3 national e-infrastructures: CESNET, CERIT-SC and IT4Innovations. The project provides ICT services mostly for Czech universities and research institutions. High energy physics projects belong to the most demanding users in terms of network, computing and storage capacities. We will describe usage of these three main categories for several HEP and astroparticle physics projects in which Czech institutions are involved. But not only these most visible services are provided. Other services like security CSIRT team, backup solutions, virtualization platform, identity management, Jupyter notebooks, electronic logbook and user support system are extensively used. Many concrete examples will be given on e-INFRA services usage by the Czech WLCG Tier-2 center. We will also present the important role of the e-INFRA project as a national coordinator of international activities in the grid computing (EGI), open science solutions (EOSC) or high-performance computing (EuroHPC)

Polymerisation and surface modification of methacrylate monoliths in polyimide channels and polyimide coated capillaries using 660 nm light emitting diodes

An investigation into the preparation of monolithic separation media utilising a cyanine dye sensitiser/triphenylbutylborate/N-methoxy-4-phenylpyridinium tetrafluoroborate initiating system activated by 660nm light emitting diodes is reported. The work demonstrates multiple uses of red- light initiated polymerisation in the preparation of monolithic stationary phases within polyimide and polyimide coated channels and the modification of monolithic materials with molecules which absorb strongly in the UV region. This initiator complex was used to synthesise poly(butyl methacrylate- co-ethylene dimethacrylate) and poly(methyl methacrylate-co-ethylene dimethacrylate) monolithic stationary phases in polyimide coated fused silica capillaries of varying internal diameters, as well as within polyimide micro-fluidic chips. The repeatability of the preparation procedure and resultant mono- lithic structure was demonstrated with a batch of poly(butyl methacrylate-co-ethylene dimethacrylate) monoliths in 10

EOSC-SYNERGY EU Deliverable D2.3: Final report on EOSC integration

This report is delivered in the form of a "Handbook" on how to integrate national clouds, thematic resources, and data repositories conformant to common quality standards, and harmonised in terms of technological, policy, and legal aspects.EOSC-SYNERGY receives funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 857647.Peer reviewe

SNPs in the FCER1A Gene Region Show No Association with Allergic Rhinitis in a Han Chinese Population

Background: Immunoglobulin E (IgE) is a central player in the allergic response, and raised total IgE levels are considered as an indicator of atopy or potential development of atopy. A recent genome-wide scan in a German population-based cohort of adults identified the gene encoding the alpha chain of the high affinity receptor for IgE (FCER1A) as a susceptibility locus influencing total serum IgE levels. The aim of this study was to investigate whether the polymorphisms in the FCER1A gene are associated with allergic rhinitis (AR) in a Han Chinese population. Methodology/Principal Findings: A population of 378 patients with AR and 288 healthy controls was studied. Precise phenotyping of patients was accomplished by means of a questionnaire and clinical examination. Blood was drawn for DNA extraction and total serum immunoglobulin E (IgE) measurement. A total of 16 single nucleotide polymorphisms (SNPs) in FCER1A were selected and individually genotyped. None of the SNPs in the FCER1A showed an association with AR. Similarly, the lack of association was also evident in subgroup analysis for the presence of different allergen sensitivities. None of the selected SNPs in FCER1A was associated with total IgE level. Conclusions: Although FCER1A presents itself as a good candidate for contributing to total serum IgE, this study failed t

Is High Resolution Melting Analysis (HRMA) Accurate for Detection of Human Disease-Associated Mutations? A Meta Analysis

BACKGROUND: High Resolution Melting Analysis (HRMA) is becoming the preferred method for mutation detection. However, its accuracy in the individual clinical diagnostic setting is variable. To assess the diagnostic accuracy of HRMA for human mutations in comparison to DNA sequencing in different routine clinical settings, we have conducted a meta-analysis of published reports. METHODOLOGY/PRINCIPAL FINDINGS: Out of 195 publications obtained from the initial search criteria, thirty-four studies assessing the accuracy of HRMA were included in the meta-analysis. We found that HRMA was a highly sensitive test for detecting disease-associated mutations in humans. Overall, the summary sensitivity was 97.5% (95% confidence interval (CI): 96.8-98.5; I(2) = 27.0%). Subgroup analysis showed even higher sensitivity for non-HR-1 instruments (sensitivity 98.7% (95%CI: 97.7-99.3; I(2) = 0.0%)) and an eligible sample size subgroup (sensitivity 99.3% (95%CI: 98.1-99.8; I(2) = 0.0%)). HRMA specificity showed considerable heterogeneity between studies. Sensitivity of the techniques was influenced by sample size and instrument type but by not sample source or dye type. CONCLUSIONS/SIGNIFICANCE: These findings show that HRMA is a highly sensitive, simple and low-cost test to detect human disease-associated mutations, especially for samples with mutations of low incidence. The burden on DNA sequencing could be significantly reduced by the implementation of HRMA, but it should be recognized that its sensitivity varies according to the number of samples with/without mutations, and positive results require DNA sequencing for confirmation

Robust open cellular porous polymer monoliths made from cured colloidal gels of latex particles

The coagulation of oppositely charged latexes, prepared from the soap-free emulsion polymerisation of styrene using water as the reaction medium, resulted in the obtainment of colloidal gels that were porous in nature and held together by electrostatic interactions. Chemical crosslinking, involving the introduction of a water-soluble crosslinker, resulted in the obtainment of stronger chemical bonds between particles affording a rigid porous material known as a monolith. It was found that, in a simpler approach, these materials could be prepared using a single latex where the addition of ammonium persulfate both resulted in the formation of the colloidal gel and initiated the crosslinking process. The pore size of the resulting monoliths was predictable as this was observed to directly correlate to the particle diameter, with larger pores achieved using particles of increased size. All gels obtained in this work were highly mouldable and retained their shape, which allowed for a range of formats to be easily prepared without the requirement of a mould