APOB CRISPR-Cas9 Engineering in Hypobetalipoproteinemia: A Promising Tool for Functional Studies of Novel Variants

Hypobetalipoproteinemia is characterized by LDL-cholesterol and apolipoprotein B (apoB) plasma levels below the fifth percentile for age and sex. Familial hypobetalipoproteinemia (FHBL) is mostly caused by premature termination codons in the APOB gene, a condition associated with fatty liver and steatohepatitis. Nevertheless, many families with a FHBL phenotype carry APOB missense variants of uncertain significance (VUS). We here aimed to develop a proof-of-principle experiment to assess the pathogenicity of VUS using the genome editing of human liver cells. We identified a novel heterozygous APOB-VUS (p.Leu351Arg), in a FHBL family. We generated APOB knock-out (KO) and APOB-p.Leu351Arg knock-in Huh7 cells using CRISPR-Cas9 technology and studied the APOB expression, synthesis and secretion by digital droplet PCR and ELISA quantification. The APOB expression was decreased by 70% in the heterozygous APOB-KO cells and almost abolished in the homozygous-KO cells, with a consistent decrease in apoB production and secretion. The APOB-p.Leu351Arg homozygous cells presented with a 40% decreased APOB expression and undetectable apoB levels in cellular extracts and supernatant. Thus, the p.Leu351Arg affected the apoB secretion, which led us to classify this new variant as likely pathogenic and to set up a hepatic follow-up in this family. Therefore, the functional assessment of APOB-missense variants, using gene-editing technologies, will lead to improvements in the molecular diagnosis of FHBL and the personalized follow-up of these patients.     

Influence of the alkyl-chain size on the amphiphilic behaviour of poly(3-alkyl-1-vinylimidazolium) bromide in aqueous medium

Summary Several 3-alkyl-1-vinylimidazolium bromides differing by the length of the alkyl chain were prepared and polymerized using a free radical route. The behaviour of the obtained homopolymers in aqueous medium was investigated by several physico-chemical methods including viscosimetry, surface tension measurements and fluorescence spectroscopy. These experiments highlighted a strong influence of the length of the pendent alkyl chain on the side-chain aggregation tendency which is a property of such amphiphilic comb polymers. Fluorescence experiments were also conducted on the surface active analogues, in order to compare their behaviour to that of the polymers. Received: 28 June 2000/Revised version: 28 November 2000/Accepted: 20 December 2000     

The impact of polypropylene‐graft‐maleic anhydride on the crystallization and dynamic mechanical properties of isotactic polypropylene

Differential scanning calorimetry (DSC) and dynamic mechanical analysis (DMA) were used to identify the mechanisms that lead to differences in the mechanical behavior of formulations of polypropylene blended with maleated polypropylene (MAPP) copolymers. MAPP lowered the melting temperature of PP indicating that less stable crystals were formed possibly because of cocrystallization of PP and MAPP. Crystallization kinetics revealed that copolymers do not change the rate of crystal growth, but may retard nucleation leading to a more spherulitic morphology. The dynamic storage modulus slightly increased in the glassy region with the small addition amounts of MAPP, while mechanical dampening systematically decreased with MAPP addition. An analysis of the viscoelastic behavior did not reveal any real differences in molecular coupling around the β‐transition of PP with the addition of the MAPP copolymer. At low addition levels, MAPP does not appear to have a significant impact on the viscoelastic properties of the polymer blend. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Hydrolysis of biliary phospholipids in the upper small intestine of the chick

Chick endogenous phospholipids were doubly labeled by an intravenous injection of [³²P] phosphate and [1-¹⁴C] oleic acid, and the free fatty acid and phospholipid fraction of gall bladder bile and in contents of upper small intestine were analyzed 4 days later. There was evidence of hydrolysis of biliary phosphatidylcholine to lysophosphatidylcholine in the duodenum and jejunum, but this did not account for the pronounced increase in the¹⁴C radioactivity of the free fatty acids relative to the³²P phospholipid radioactivity between bile and upper intestinal segments. It is suggested that phosphatidylcholine is largely absorbed in the duodenum of the chick while the remainder is progressively hydrolyzed and absorbed.     

COVID-19 Pandemic: Escape of Pathogenic Variants and MHC Evolution

We propose a new hypothesis that explains the maintenance and evolution of MHC polymorphism. It is based on two phenomena: the constitution of the repertoire of naive T lymphocytes and the evolution of the pathogen and its impact on the immune memory of T lymphocytes. Concerning the latter, pathogen evolution will have a different impact on reinfection depending on the MHC allomorph. If a mutation occurs in a given region, in the case of MHC allotypes, which do not recognize the peptide in this region, the mutation will have no impact on the memory repertoire. In the case where the MHC allomorph binds to the ancestral peptides and not to the mutated peptide, that individual will have a higher chance of being reinfected. This difference in fitness will lead to a variation of the allele frequency in the next generation. Data from the SARS-CoV-2 pandemic already support a significant part of this hypothesis and following up on these data may enable it to be confirmed. This hypothesis could explain why some individuals after vaccination respond less well than others to variants and leads to predict the probability of reinfection after a first infection depending upon the variant and the HLA allomorph.     

智能蓝光捕捉技术简述

为了防止有害光线对眼部的伤害,依视路集团研发出了智能蓝光捕捉技术,该技术可实现紫外线防护,符合新国标要求的蓝光防护,并维持色彩真实性。     

Megaprojects as complex adaptive systems: The Hinkley point C case

Megaprojects are complex projects which impact millions of people, involve public and private stakeholders, and present challenges related to decision making and performance shortfalls. They are relevant cases for studying faulty management thinking as well as performance evolutions and self-organizing dynamics. Our paper builds on the theory of Complex Adaptive Systems (CAS) to understand and model processes of evolution in the Hinkley Point C nuclear power plant megaproject. The results show that CAS properties apply to megaproject changes and provide a theoretical and practical framework for examining and modeling megaproject management dynamics. We designed a research methodology combining content analysis and historical research for its relevance in conducting organizational research in conditions of complexity and non-linearity. This original research design makes it possible to conduct causal analyses of relations between key megaproject events and thus build models of evolution dynamics in stakeholder success expectations, change mechanisms in the implementation of project outputs, and self-organizing patterns.     

Immunocytochemical localisation of para-coumaric acid and feruloyl-arabinose in the cell walls of maize stem

Two phenolic compounds, p-coumaric acid and feruloyl-arabinose, were localised by immunocytochemistry in the cell walls of the apical internode of two lines of maize (Co125 and W401) of different digestibility. The compounds were detected at two stages of cell maturity in the lignified tissues (sclerenchyma, fibres and xylem) and in the medullary parenchyma, which, in the samples studied, was not lignified. p-Coumaric acid is a phenolic acid associated with lignins, which confer resistance on plant cell walls to microbial degradation in the rumen. Feruloyl-arabinose is a compound associated with xylans, the principal hemicelluloses in Gramineae, which are potentially degradable. Labelling of p-coumaric acid decreased in both maize lines with cell age and as the cell walls became lignified. The mass of lignin deposited in the cell walls masked p-coumaric acid, thereby making it less accessible to the antibodies. There was an inverse relationship in the labelling of p-coumaric acid and feruloyl-arabinose. Feruloyl-arabinose was more heavily labelled as the plant cell walls matured in all the lignified tissues of both maize lines and in the parenchyma of the less digestible line. All tissues except the parenchyma were more heavily labelled with both sera in Co125, the more digestible line. © 1998 Society of Chemical Industry.     

The evolution of the exhaust diesel soot adsorptive properties after NO2 exposure and according to their SOF content

Several samples of exhaust diesel soot are investigated by inverse gas chromatography and linear solvation energy relationship (LSER) modelling according to their soluble organic fraction content and their time of exposure in oxidative conditions. The results demonstrate the evolution of the adsorptive properties of the studied materials towards volatile compounds during the oxidation under NO₂.