Selective Cardiomyocyte Oxidative Stress Leads to Bystander Senescence of Cardiac Stromal Cells

Accumulation of senescent cells in tissues during normal or accelerated aging has been shown to be detrimental and to favor the outcomes of age-related diseases such as heart failure (HF). We have previously shown that oxidative stress dependent on monoamine oxidase A (MAOA) activity in cardiomyocytes promotes mitochondrial damage, the formation of telomere-associated foci, senescence markers, and triggers systolic cardiac dysfunction in a model of transgenic mice overexpressing MAOA in cardiomyocytes (Tg MAOA). However, the impact of cardiomyocyte oxidative stress on the cardiac microenvironment in vivo is still unclear. Our results showed that systolic cardiac dysfunction in Tg MAOA mice was strongly correlated with oxidative stress induced premature senescence of cardiac stromal cells favoring the recruitment of CCR2⁺ monocytes and the installation of cardiac inflammation. Understanding the interplay between oxidative stress induced premature senescence and accelerated cardiac dysfunction will help to define new molecular pathways at the crossroad between cardiac dysfunction and accelerated aging, which could contribute to the increased susceptibility of the elderly to HF.     

Light propagation in multilayered scattering media beyond the diffusive regime

We describe a method to solve the radiative transfer equation (RTE) in multilayered geometry with index mismatch and demonstrate its potential for modeling light propagation in biological systems. The method is compared to Monte Carlo simulations with high accuracy but is much more efficient in terms of computer time. We illustrate the potential of the method by studying a multilayered system containing a weakly scattering layer surrounded by highly scattering layers, with anisotropic scattering and index mismatched interfaces. The calculation of directional transmitted fluxes has shown that the RTE method can be used to calculate relevant quantities in realistic systems in the presence of non-diffusive behavior.     

Analysis of Amyloid-β Peptides in Cerebrospinal Fluid Samples by Capillary Electrophoresis Coupled with LIF Detection

We report a CE-LIF method for the separation and detection of five synthetic amyloid-β peptides corresponding to an important family of CSF-biomarkers in the context of Alzheimer disease (AD). The presumed most relevant peptides (Aβ1-42, Aβ1-40, and Aβ1-38) that may support the differentiation between AD and healthy patients or other dementias were successfully detected in CSF by incorporating an immunoconcentration step prior to CE analysis of derivatized peptides. We labeled the Aβ peptides with a fluoroprobe dye before CE-LIF analysis. This reagent reacts with the amino groups of lysine residues and produced mostly ditagged Aβ peptides under the proposed experimental conditions. The labeling reaction displayed similar efficiency with each one of the five different synthetic Aβ peptides that were tested. The limit of detection of the CE-LIF method approached 280 attomoles of injected synthetic labeled Aβ peptides. We obtained excellent correlation between peak areas and peptide concentrations from 35 nM to 750 nM. For the detection of Aβ peptides in human CSF samples, we enriched the peptides by immunoprecipitation prior to the CE-LIF analysis. The comparison of the CE-LIF profiles obtained from CSF samples from 3 AD patients and 4 non-demented control subjects indicated noticeable differences, suggesting that this method, which relies on a multibiomarker approach, may have potential as a clinical diagnostic test for AD.     

NMR-based structural glycomics for high-throughput screening of carbohydrate-active enzyme specificity

We report here the development of a straightforward, sensitive, and quantitative NMR-based method for high-throughput characterization of carbohydrate structure and screening of carbohydrate active enzyme (CAZyme) specificity. Automated assays starting from gene library expression to carbohydrate structure determination directly from crude reaction media have been established and successfully used to screen a library of 4032 CAZymes obtained by combinatorial engineering, at a rate of 480 enzyme variants per day. This allowed one to accurately discriminate 303 enzyme variants with altered specificity. The results demonstrate the potential of high-throughput NMR technology in glycomics, to mine artificial and natural enzyme diversity for novel biocatalysts.     

Förster resonance energy transfer imaging in vivo with approximated radiative transfer equation

We describe a new light transport model, which was applied to three-dimensional lifetime imaging of F?rster resonance energy transfer in mice in vivo. The model is an approximation to the radiative transfer equation and combines light diffusion and ray optics. This approximation is well adopted to wide-field time-gated intensity-based data acquisition. Reconstructed image data are presented and compared with results obtained by using the telegraph equation approximation. The new approach provides improved recovery of absorption and scattering parameters while returning similar values for the fluorescence parameters.     

Nonlinear plasmonics at planar metal surfaces

We investigate the nonlinear optical response of a noble metal surface. We derive the components of the third-order nonlinear susceptibility and determine an absolute value of χ((3))≈0.2 nm(2) V(-2), a value that is more than two orders of magnitude larger than the values found for typical nonlinear laser crystals. Using nonlinear four-wave mixing (4WM) with incident laser pulses of frequencies ω(1) and ω(2), we generate fields oscillating at the nonlinear frequency ω(4WM)=2ω(1)-ω(2). We identify and discuss three distinct regimes: (i) a regime where the 4WM field is propagating, (ii) a regime where it is evanescent, and (iii) a regime where the nonlinear response couples to surface plasmon polaritons.     

Statistical analysis of effects of industrial processing steps on functional properties of pasteurised liquid egg white

Egg white is widely used as an ingredient in the food industry owing to its excellent functional properties. The transformation of shell eggs into safe liquid, frozen, or spray‐dried egg white with extended shelf life requires many technological operations that result in modifications to the egg white's functional properties. The present study highlights the critical steps affecting foaming and gelling properties during a classical pasteurised liquid egg white process. The main source of variation in functional properties was raw material quality, accounting for 70% of the variability. Part of the remaining 30% was explained by mechanical egg white–yolk separation, tank storage, pasteurisation and homogenisation that resulted in damage to the functional properties, whereas initial flow through pipes and pumping resulted in their improvement. The effects of these steps could be grouped according to the type of treatment undertaken. Dry matter content, pH and treatment intensity at each step contributed about 30% of the variability in functional properties due to processing steps. Relationships between the modifications of egg white functional properties and protein conformation were established. Between 46 and 78% of the variability in functional properties can be explained by protein denaturation, temperature and enthalpy changes. Copyright © 2004 Society of Chemical Industry     

Towards Molecular Understanding of the Functional Role of UbiJ-UbiK2 Complex in Ubiquinone Biosynthesis by Multiscale Molecular Modelling Studies

Ubiquinone (UQ) is a polyisoprenoid lipid found in the membranes of bacteria and eukaryotes. UQ has important roles, notably in respiratory metabolisms which sustain cellular bioenergetics. Most steps of UQ biosynthesis take place in the cytosol of E. coli within a multiprotein complex called the Ubi metabolon, that contains five enzymes and two accessory proteins, UbiJ and UbiK. The SCP2 domain of UbiJ was proposed to bind the hydrophobic polyisoprenoid tail of UQ biosynthetic intermediates in the Ubi metabolon. How the newly synthesised UQ might be released in the membrane is currently unknown. In this paper, we focused on better understanding the role of the UbiJ-UbiK₂ heterotrimer forming part of the metabolon. Given the difficulties to gain functional insights using biophysical techniques, we applied a multiscale molecular modelling approach to study the UbiJ-UbiK₂ heterotrimer. Our data show that UbiJ-UbiK₂ interacts closely with the membrane and suggests possible pathways to enable the release of UQ into the membrane. This study highlights the UbiJ-UbiK₂ complex as the likely interface between the membrane and the enzymes of the Ubi metabolon and supports that the heterotrimer is key to the biosynthesis of UQ₈ and its release into the membrane of E. coli.     

TNF-α and IL-1β Modulate Blood-Brain Barrier Permeability and Decrease Amyloid-β Peptide Efflux in a Human Blood-Brain Barrier Model

The blood-brain barrier (BBB) is a selective barrier and a functional gatekeeper for the central nervous system (CNS), essential for maintaining brain homeostasis. The BBB is composed of specialized brain endothelial cells (BECs) lining the brain capillaries. The tight junctions formed by BECs regulate paracellular transport, whereas transcellular transport is regulated by specialized transporters, pumps and receptors. Cytokine-induced neuroinflammation, such as the tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), appear to play a role in BBB dysfunction and contribute to the progression of Alzheimer’s disease (AD) by contributing to amyloid-β (Aβ) peptide accumulation. Here, we investigated whether TNF-α and IL-1β modulate the permeability of the BBB and alter Aβ peptide transport across BECs. We used a human BBB in vitro model based on the use of brain-like endothelial cells (BLECs) obtained from endothelial cells derived from CD34+ stem cells cocultivated with brain pericytes. We demonstrated that TNF-α and IL-1β differentially induced changes in BLECs’ permeability by inducing alterations in the organization of junctional complexes as well as in transcelluar trafficking. Further, TNF-α and IL-1β act directly on BLECs by decreasing LRP1 and BCRP protein expression as well as the specific efflux of Aβ peptide. These results provide mechanisms by which CNS inflammation might modulate BBB permeability and promote Aβ peptide accumulation. A future therapeutic intervention targeting vascular inflammation at the BBB may have the therapeutic potential to slow down the progression of AD.     

A Novel Thermoplastic Composite for Marine Applications: Comparison of the Effects of Aging on Mechanical Properties and Diffusion Mechanisms

Applied Composite Materials - This article investigates and compares the effects of hydrothermal aging on carbon fibre / Elium? thermoplastic composite and on carbon fibre / vinylester...