Synthesis of C‐Glucosylated Octaketide Anthraquinones in Nicotiana benthamiana by Using a Multispecies‐Based Biosynthetic Pathway

Carminic acid is a C‐glucosylated octaketide anthraquinone and the main constituent of the natural dye carmine (E120), possessing unique coloring, stability, and solubility properties. Despite being used since ancient times, longstanding efforts to elucidate its route of biosynthesis have been unsuccessful. Herein, a novel combination of enzymes derived from a plant (Aloe arborescens, Aa), a bacterium (Streptomyces sp. R1128, St), and an insect (Dactylopius coccus, Dc) that allows for the biosynthesis of the C‐glucosylated anthraquinone, dcII, a precursor for carminic acid, is reported. The pathway, which consists of AaOKS, StZhuI, StZhuJ, and DcUGT2, presents an alternative biosynthetic approach for the production of polyketides by using a type III polyketide synthase (PKS) and tailoring enzymes originating from a type II PKS system. The current study showcases the power of using transient expression in Nicotiana benthamiana for efficient and rapid identification of functional biosynthetic pathways, including both soluble and membrane‐bound enzymes.     

Model-based multivariable control of the drying of a thin sheet of fibres in a continuous infrared dryer

We developed an Internal Model Control (IMC) algorithm for drying a thin textile fabric, continuously passing through an electric infrared dryer, based on a reduced linear model of the drying dynamics. This model relates the controlled variables, the humidity and temperature of the web at the dryer outlet, to the manipulated variables, the electrical power supplied to the sources and the web speed through the dryer, and also to changes in the initial humidity of the web at the dryer inlet. The control algorithm was first tested by simulation using the model in regulation mode, and in set-point tracking mode, to vary the manipulated variables so as to maintain the controlled variables at their respective set-points when the inlet web temperature and humidity were changed. The performance under simulated operational conditions was compared to that of a conventional feedback proportional-integral (PI) controller coupled with a feedforward control. The IMC controller was then tested directly in regulation mode using a pilot scale infrared dryer, acting simultaneously on the manipulated variables, the emitter power and the web speed, to control the fabric temperature and humidity at the dryer outlet. The experimental results were compared with those from the above feedback-feedforward controller, on the pilot scale dryer. The results have indicated that the closed-loop stability of the process is assured simply by choosing a stable IMC controller. Also, such a controller does not require the design of specific compensators for the strong interactions between variables of the drying process.     

Simulation of Fluctuating Populations of Microorganisms and Macroorganisms with Models Having a Normal Random Variate Term

Fluctuating populations of micro- and macroorganisms play a major role in the stability and safety of food and agricultural products. In recent years such populations have been described by ‘chaotic’ models, most notably the discrete logistic function. This model's major limitations, however, are that it implies determinism and that it can account for neither population explosions nor extinction. Consequently, it is suggested that in order to simulate realistic population histories, the growth rate constant, r, in the original discrete logistic function X_n+1=rX_n(1-X_n) can be replaced by a randomly varying term, r_n. The latter can be defined as r_n=r₀ exp(kZ_rn) where r₀ is a characteristic constant, Z_rn a normal random variate produced by a normally distributed random number generator and k its chosen standard deviation. Theoretically, the magnitude of such r_n (and its corresponding X_n) can be anywhere in the range 0<X_n<∞. However, the magnitude of k can be selected so that X_n>1 is a very rare event. In the special case of widely fluctuating populations dominated by random factors, the model can be replaced by N_n/N₀=exp(k′Z_rn), where N₀ is a characteristic number. In this form the model can account for occasional aperiodic population explosions, and if a lower limit to a viable number is also set (eg N/N₀>0·01), for extinction events as well. The flexibility of the two models is demonstrated with simulated population evolution patterns of different kinds. © 1997 SCI.     

Impact of Host Immune Status on Discordant Anti-SARS-CoV-2 Circulating B Cell Frequencies and Antibody Levels

Individuals with pre-existing chronic systemic low-grade inflammation are prone to develop severe COVID-19 and stronger anti-SARS-CoV-2 antibody responses. Whether this phenomenon reflects a differential expansion of antiviral B cells or a failure to regulate antibody synthesis remains unknown. Here, we compared the antiviral B cell repertoire of convalescent healthcare personnel to that of hospitalized patients with pre-existing comorbidities. Out of 277,500 immortalized B cell clones, antiviral B cell frequencies were determined by indirect immunofluorescence screening on SARS-CoV-2 infected cells. Surprisingly, frequencies of SARS-CoV-2 specific clones from the two groups were not statistically different, despite higher antibody levels in hospitalized patients. Moreover, functional analyses revealed that several B cell clones from healthcare personnel with low antibody levels had neutralizing properties. This study reveals for the first time a key qualitative defect of antibody synthesis in severe patients and calls for caution regarding estimated protective immunity based only on circulating antiviral antibodies.     

Exploring the early steps of amyloid peptide aggregation by computers

The assembly of normally soluble proteins into amyloid fibrils is a hallmark of neurodegenerative diseases. Because protein aggregation is very complex, involving a variety of oligomeric metastable intermediates, the detailed aggregation paths and structural characterization of the intermediates remain to be determined. Yet, there is strong evidence that these oligomers, which form early in the process of fibrillogenesis, are cytotoxic. In this paper, we review our current understanding of the underlying factors that promote the aggregation of peptides into amyloid fibrils. We focus here on the structural and dynamic aspects of the aggregation as observed in state-of-the-art computer simulations of amyloid-forming peptides with an emphasis on the activation-relaxation technique.     

Derivation of a control oriented low order model of the dynamics of an infrared oven for the continuous drying of a thin sheet of material

A model consisting of three partial non-linear differential equations for describing the humidity and temperature of a thin sheet of material and of the temperature of the air flow as a function of time and position in an infrared dryer was reduced to a model more adaptable to direct digital control. To begin with the original model was discretized along the length of the dryer and then linearized. Models of orders ranging from 12 to 72 were analyzed for controllability and observability. Using Moore's method, the models thus obtained were further reduced to a controllable state space model of order 5 that produced satisfactory results and for which stability was preserved. Simulated open loop responses of the reduced model to discrete perturbations in emitter electrical power, inlet humidity and velocity of the web, produced satisfactory dynamic and steady-state responses when compared to responses of the original model subjected to similar perturbations. The steady-state responses showed absolute errors for humidity and temperature of the web less than 1.3% [g water/100 g] and 2.2°C, respectively.     

Thin chromium nitride PVD coatings on stainless steel for conductive component as bipolar plates of PEM fuel cells: Ex-situ and in-situ performances evaluation

In this paper, two types of chromium PVD coatings (100 nm) have been elaborated on 316L stainless steel (SS) by adjusting the nitrogen flow rate. The first coating is a mixture of Cr₂N and Cr, the second one is a single phase CrN. It is shown that the performances of the material are strongly dependant of the nature of the passive film formed on the chromium nitride layers due to the galvanic coupling between the coating and the substrate. The CrN coated SS shows very good corrosion resistance in simulated PEMFC media. The surface conductivity of the SS is also greatly improved and the CrN coated SS shows an interfacial contact resistance of 10 mΩ cm² at 140 N cm⁻². Five single cells of stainless steel bipolar plates coated with the CrN film were assembled for performance test. This 5 cell stack does not show any mean voltage degradation over 200 h dynamic cycling. Moreover, the performances of the CrN coated SS bipolar plates are very close to the Au-coated SS bipolar plates.     

Synthesis of conducting transparent few-layer graphene directly on glass at 450 °C

Post-growth transfer and high growth temperature are two major hurdles that research has to overcome to get graphene out of research laboratories. Here, using a plasma-enhanced chemical vapour deposition process, we demonstrate the large-area formation of continuous transparent graphene layers at temperatures as low as 450?°C. Our few-layer graphene grows at the interface between a pre-deposited 200 nm Ni catalytic film and an insulating glass substrate. After nickel etching, we are able to measure the optical transmittance of the layers without any transfer. We also measure their sheet resistance directly and after inkjet printing of electrical contacts: sheet resistance is locally as low as 500 Ω sq?1. Finally the samples equipped with printed contacts appear to be efficient humidity sensors.     

Cerium insertion in 316L passive film: Effect on conductivity and corrosion resistance performances of metallic bipolar plates for PEM fuel cell application

A surface treatment to modify passive film properties formed on 316L stainless steel has been designed for bipolar plate application in PEMFC systems. This one is based on cerium insertion from electrochemical technique. The effect of this element addition on the composition of the passive films was evaluated by X-ray photoelectron spectroscopy (XPS) and low energy ion spectroscopy (LEIS). Potentiodynamic and potentiostatic curves, and electrochemical impedance spectroscopy (EIS) were used for the characterisation of the corrosion protection in simulated PEMFC media. Interfacial contact resistance (ICR) was also evaluated. Results highlight an increase of the polarisation resistance of the cerium treated sample at ambient temperature, and a good corrosion resistance in anodic and cathodic simulated media. An important diminution of the IRC is obtained (80% with respect to the reference), which is attributed to the enhancement of the passive layer conductivity of the processed sample. Surface analyses show a great enrichment of cerium in first atomic layers of the films, which explains the better conductivity observed by an increase of charge carrier density in the passive film.     

Modulation of Starch Digestibility in Breakfast Cereals Consumed by Subjects with Metabolic Risk: Impact on Markers of Oxidative Stress and Inflammation during Fasting and the Postprandial Period

Scope : Decreasing postprandial glycaemic excursions may have a beneficial effect on inflammatory and oxidative stress profiles. In this study, we investigated the impact of carbohydrate digestibility modulation per se, as a means of reducing the glycaemic response, on metabolic and inflammatory responses in subjects with metabolic risk factors. Methods and results : Twenty healthy subjects with metabolic risk consumed a cereal product either high in Slowly Digestible Starch (HSDS) or low in SDS (LSDS) at breakfast daily for 3 weeks, in a cross‐over design. Following each 3‐week session, postprandial glycaemia, insulinaemia, the lipid profile, inflammation and oxidative stress markers were assessed and compared to those induced by ingestion of a glucose solution (as a reference). The 2‐h glycaemic and insulinaemic responses were significantly lower following the HSDS breakfast compared with the LSDS breakfast or glucose. No significant differences between the products were observed in terms of the lipid profile, C‐reactive protein, IL‐6 and tumour necrosis factor alpha. We observed a slight increase in fasting lipid peroxidation markers, including an increase in plasma malondialdehyde (MDA) and a decrease in whole blood glutathione (GSH), without significant alteration of urinary F2‐isoprostanes or plasma glutathione peroxidase (GPx) activity. Conclusion : Consumption of HSDS products for 3 weeks significantly altered both postprandial glycaemia and insulinaemia, but was not sufficient to modify the inflammatory profile. Consumption of both cereal products was associated with a slightly higher fasting oxidative stress profile.