RAB10 Interacts with ABCB4 and Regulates Its Intracellular Traffic

ABCB4 (ATP-binding cassette subfamily B member 4) is an ABC transporter expressed at the canalicular membrane of hepatocytes where it ensures phosphatidylcholine secretion into bile. Genetic variations of ABCB4 are associated with several rare cholestatic diseases. The available treatments are not efficient for a significant proportion of patients with ABCB4-related diseases and liver transplantation is often required. The development of novel therapies requires a deep understanding of the molecular mechanisms regulating ABCB4 expression, intracellular traffic, and function. Using an immunoprecipitation approach combined with mass spectrometry analyses, we have identified the small GTPase RAB10 as a novel molecular partner of ABCB4. Our results indicate that the overexpression of wild type RAB10 or its dominant-active mutant significantly increases the amount of ABCB4 at the plasma membrane expression and its phosphatidylcholine floppase function. Contrariwise, RAB10 silencing induces the intracellular retention of ABCB4 and then indirectly diminishes its secretory function. Taken together, our findings suggest that RAB10 regulates the plasma membrane targeting of ABCB4 and consequently its capacity to mediate phosphatidylcholine secretion.     

The relation between (bio-)chemical, morphological, and mechanical properties of thermally processed carrots as influenced by high-pressure pretreatment condition

Thermal texture evolution kinetics (90–110 °C) of nonpretreated and high-pressure pretreated (HP = 400 MPa, 60 °C, 15 min) carrots were determined using a multiparameter approach (cutting, compression). Alcohol Insoluble Residues (AIR) were extracted before and after thermal processing of the samples and the degree of methylation (DM) was estimated. The β-elimination kinetics of the water soluble pectin extract from the AIR was studied and related to the changes in material properties. Morphological changes and tissue-failure characteristics were monitored. The mechanical properties were strongly dependent on the processing temperature and the DM of the samples. Texture degradation rate constants were independent of the texture measurement method. Increasing temperature accelerated the β-elimination reaction (k _b) and the thermosoftening (k _x) rates, but pretreatment condition slowed down the rates. Interestingly, a strong correlation (r > 0.99) between k _b and k _x occurred. Thermal processing resulted in cell-wall thickening accompanied by a transition from cell rupture to cell separation, a process retarded by pretreatment condition.     

Thyroid Hormones and Antioxidant Systems: Focus on Oxidative Stress in Cardiovascular and Pulmonary Diseases

In previous works we demonstrated an inverse correlation between plasma Coenzyme Q₁₀ (CoQ₁₀) and thyroid hormones; in fact, CoQ₁₀ levels in hyperthyroid patients were found among the lowest detected in human diseases. On the contrary, CoQ₁₀ is elevated in hypothyroid subjects, also in subclinical conditions, suggesting the usefulness of this index in assessing metabolic status in thyroid disorders. A Low-T3 syndrome is a condition observed in several chronic diseases: it is considered an adaptation mechanism, where there is a reduction in pro-hormone T4 conversion. Low T3-Syndrome is not usually considered to be corrected with replacement therapy. We review the role of thyroid hormones in regulation of antioxidant systems, also presenting data on total antioxidant capacity and Coenzyme Q₁₀. Published studies suggest that oxidative stress could be involved in the clinical course of different heart diseases; our data could support the rationale of replacement therapy in low-T3 conditions.     

An E3‐14.7K Peptide that Promotes Microtubules‐Mediated Transport of Plasmid DNA Increases Polyplexes Transfection Efficiency

Chemical vectors as cationic polymers and cationic lipids are promising alternatives to viral vectors for gene therapy. Beside endosome escape and nuclear import, plasmid DNA (pDNA) migration in the cytosol toward the nuclear envelope is also regarded as a limiting step for efficient DNA transfection with non‐viral vectors. Here, the interaction between E3‐14.7K and FIP‐1 to favor migration of pDNA along microtubules is exploited. E3‐14.7K is an early protein of human adenoviruses that interacts via FIP‐1 (Fourteen.7K Interacting Protein 1) protein with the light‐chain components of the human microtubule motor protein dynein (TCTEL1). This peptide is conjugated with pDNA and mediates interaction of pDNA in vitro with isolated microtubules as well as with microtubules in cellulo. Videomicroscopy and tracking treatment of images clearly demonstrate that P79‐98/pDNA conjugate exhibits a linear transport with large amplitude along microtubules upon 2 h transfection with polyplexes whereas control pDNA conjugate exhibits small non‐directional movements in the cytoplasm. Remarkably, P79‐98/peGFP polyplexes enhance by a factor 2.5 (up to 76%) the number of transfected cells. The results demonstrate, for the first time, that the transfection efficiency of polyplexes can be drastically increased when the microtubules migration of pDNA is facilitated by a peptide allowing pDNA docking to TCTEL1. This is a real breakthrough in the non viral gene delivery field that opens hope to build artificial viruses.     

Pre-Omicron Vaccine Breakthrough Infection Induces Superior Cross-Neutralization against SARS-CoV-2 Omicron BA.1 Compared to Infection Alone

SARS-CoV-2 variants raise concern because of their high transmissibility and their ability to evade neutralizing antibodies elicited by prior infection or by vaccination. Here, we compared the neutralizing abilities of sera from 70 unvaccinated COVID-19 patients infected before the emergence of variants of concern (VOCs) and of 16 vaccine breakthrough infection (BTI) cases infected with Gamma or Delta against the ancestral B.1 strain, the Gamma, Delta and Omicron BA.1 VOCs using live virus. We further determined antibody levels against the Nucleocapsid (N) and full Spike proteins, the receptor-binding domain (RBD) and the N-terminal domain (NTD) of the Spike protein. Convalescent sera featured considerable variability in the neutralization of B.1 and in the cross-neutralization of different strains. Their neutralizing capacity moderately correlated with antibody levels against the Spike protein and the RBD. All but one convalescent serum failed to neutralize Omicron BA.1. Overall, convalescent sera from patients with moderate disease had higher antibody levels and displayed a higher neutralizing ability against all strains than patients with mild or severe forms of the disease. The sera from BTI cases fell into one of two categories: half the sera had a high neutralizing activity against the ancestral B.1 strain as well as against the infecting strain, while the other half had no or a very low neutralizing activity against all strains. Although antibody levels against the spike protein and the RBD were lower in BTI sera than in unvaccinated convalescent sera, most neutralizing sera also retained partial neutralizing activity against Omicron BA.1, suggestive of a better cross-neutralization and higher affinity of vaccine-elicited antibodies over virus-induced antibodies. Accordingly, the IC50: antibody level ratios were comparable for BTI and convalescent sera, but remained lower in the neutralizing convalescent sera from patients with moderate disease than in BTI sera. The neutralizing activity of BTI sera was strongly correlated with antibodies against the Spike protein and the RBD. Together, these findings highlight qualitative differences in antibody responses elicited by infection in vaccinated and unvaccinated individuals. They further indicate that breakthrough infection with a pre-Omicron variant boosts immunity and induces cross-neutralizing antibodies against different strains, including Omicron BA.1.     

Macrocell corrosion of steel in concrete: Characterization of anodic behavior in relation to the chloride content

This study presents the determination of electrochemical properties of active steel in mortar, based on inverse numerical modeling that focuses on their dependency on chloride content. An experimental campaign, consisting of galvanic coupling tests between anode samples contaminated with different chloride concentrations and cathode samples without chlorides, was carried out. Cathode polarization tests allowed for directly determining passive steel electrochemical parameters. Anode polarization tests coupled with a numerical optimization were then performed for quantifying active steel parameters and focusing on chloride's effect on the iron anodic Tafel coefficient. Furthermore, the steel electrochemical properties were successfully used as input parameters to model the galvanic experiments.     

Structure and properties of sugar beet fibres

Fibrous fractions were prepared from sugar beet pulp (RF) by sequential extractions with potassium oxalate, 0.05 M hydrochloric acid at 85°C and 0.05 m sodium hydroxide at 4°C. The overall composition, polysaccharide structure and some physico-chemical properties (cation exchange capacity, CEC; water holding capacity, WHC; swelling) of each fraction were determined. RF was mostly composed of carbohydrates (66.3 %) with minor amounts of ash, proteins and lignin. The main polysaccharides were highly methylated and acetylated pectins, cellulose and arabinans. The oxalate residue (82.1 % of RF) exhibited only minute differences from RF whereas the acidic and alkaline residues, accounting for 42.8 % and 35.5 % of RF, respectively, were enriched in cellulose and hemicelluloses (xylans, xyloglucans, mannans) and mostly devoid of pectins and arabinans. CEC and WHC of fractions were closely related to the content of unmethylated galacturonic acid residues. The influence of the ionic form of ionisable groups was demonstrated, the Na⁺ form giving the highest WHC. The ionic strength of the medium can reduce the WHC, especially in the case of the acid and alkali-extracted fibres.     

Fabrication of long-term hydrophilic surfaces of poly(dimethyl siloxane) using 2-hydroxy ethyl methacrylate

In the present work, 2-hydroxy ethyl methacrylate (HEMA) was used to modify surface of poly(dimethyl siloxane) (PDMS) elastomer. Fourier transform infrared spectroscopy (FTIR) and wetting angle measurements were used for the analysis of modified surface and hydrophilic stability of PDMS. Results of the surface reconstruction reveal that long-term hydrophilic surfaces of PDMS can be achieved by use of HEMA.