Development of a single ion hit facility at the Pierre Sue Laboratory: a collimated microbeam to study radiological effects on targeted living cells

A single ion hit facility is being developed at the Pierre Süe Laboratory (LPS) since 2004. This set-up will be dedicated to the study of ionising radiation effects on living cells, which will complete current research conducted on uranium chemical toxicity on renal and osteoblastic cells. The study of the response to an exposure to alpha particles will allow us to distinguish radiological and chemical toxicities of uranium, with a special emphasis on the bystander effect at low doses. Designed and installed on the LPS Nuclear microprobe, up to now dedicated to ion beam microanalysis, this set-up will enable us to deliver an exact number of light ions accelerated by a 3.75 MV electrostatic accelerator. An 'in air' vertical beam permits the irradiation of cells in conditions compatible with cell culture techniques. Furthermore, cellular monolayer will be kept in controlled conditions of temperature and atmosphere in order to diminish stress. The beam is collimated with a fused silica capillary tubing to target pre-selected cells. Motorisation of the collimator with piezo-electric actuators should enable fast irradiation without moving the sample, thus avoiding mechanical stress. An automated epifluorescence microscope, mounted on an antivibration table, allows pre- and post-irradiation cell observation. An ultra thin silicon surface barrier detector has been developed and tested to be able to shoot a cell with a single alpha particle.     

Temperature control during Spark Plasma Sintering and application to up-scaling and complex shaping

The determination and the homogeneity of the sample temperature during consolidation of powders by Spark Plasma Sintering (SPS) are addressed. Densifications were carried out in three different facilities differing by their constructors and their size. A structural transformation activated in a TiAl alloy was used as a marker of the sample temperature and finite element modeling were performed to evaluate the temperature at each point of the set-up. A good agreement between experimental and simulated data is exhibited. Alloys with identical microstructures were sintered and the homogeneity of the microstructure was better in the largest machines or when the sample was isolated by alumina disks.Large specimens with a diameter of 100 mm and a height of 60 mm were sintered, with the aim to achieve full densification and good microstructural control. On the basis of this temperature mastering, samples with complex shape were successfully densified. SEM post mortem observations confirmed the absence of porosities, and the expected microstructure as predicted by FEM calculations.     

Tumor Suppressive Role of miR-342-5p in Human Chondrosarcoma Cells and 3D Organoids

Chondrosarcomas are malignant bone tumors. Their abundant cartilage-like extracellular matrix and their hypoxic microenvironment contribute to their resistance to chemotherapy and radiotherapy, and no effective therapy is currently available. MicroRNAs (miRNAs) may be an interesting alternative in the development of therapeutic options. Here, for the first time in chondrosarcoma cells, we carried out high-throughput functional screening using impedancemetry, and identified five miRNAs with potential antiproliferative or chemosensitive effects on SW1353 chondrosarcoma cells. The cytotoxic effects of miR-342-5p and miR-491-5p were confirmed on three chondrosarcoma cell lines, using functional validation under normoxia and hypoxia. Both miRNAs induced apoptosis and miR-342-5p also induced autophagy. Western blots and luciferase reporter assays identified for the first time Bcl-2 as a direct target of miR-342-5p, and also Bcl-xL as a direct target of both miR-342-5p and miR-491-5p in chondrosarcoma cells. MiR-491-5p also inhibited EGFR expression. Finally, only miR-342-5p induced cell death on a relevant 3D chondrosarcoma organoid model under hypoxia that mimics the in vivo microenvironment. Altogether, our results revealed the tumor suppressive activity of miR-342-5p, and to a lesser extent of miR-491-5p, on chondrosarcoma lines. Through this study, we also confirmed the potential of Bcl-2 family members as therapeutic targets in chondrosarcomas.     

Analyzing the Substrate Specificity of a Class of Long-Horned-Beetle-Derived Xylanases by Using Synthetic Arabinoxylan Oligo- and Polysaccharides

Xylophagous long-horned beetles thrive in challenging environments. To access nutrients, they secrete plant-cell-wall-degrading enzymes in their gut fluid; among them are cellulases of the subfamily 2 of glycoside hydrolase family 5 (GH5_2). Recently, we discovered that several beetle-derived GH5_2s use xylan as a substrate instead of cellulose, which is unusual for this family of enzymes. Here, we analyze the substrate specificity of a GH5_2 xylanase from the beetle Apriona japonica (AJAGH5_2-1) using commercially available substrates and synthetic arabinoxylan oligo- and polysaccharides. We demonstrate that AJAGH5_2-1 processes arabinoxylan polysaccharides in a manner distinct from classical xylanase families such as GH10 and GH11. AJAGH5_2-1 is active on long oligosaccharides and cleaves at the non-reducing end of a substituted xylose residue (position +1) only if: 1) three xylose residues are present upstream and downstream of the cleavage site, and 2) xylose residues at positions −1, −2, +2 and +3 are not substituted.     

A multi-class approximation technique for the analysis of kanban-like control systems

Analytical methods have been proposed in the literature for performance evaluation of kanban control systems. Among them, the method presented by Di Mascolo and colleagues appears to be of special interest since it can handle manufacturing stages consisting of any number of machines and it is fairly accurate. This paper presents a new way of deriving the analytical method presented by Di Mascolo et al. The approach is to see the queueing network of the kanban control system as a multiclass queueing network in which each kanban loop is represented by a class of customers. This allows one to use the general technique proposed in Baynat and Dallery for analysing multiclass queueing network using product-form approximation methods. In terms of equations, the new method is equivalent to that previously presented. However, the computational algorithm is much more efficient since it avoids the two levels of iterations involved in the original algorithm. Another major advantage of the new method over that originally proposed is that it provides a general framework for the analysis of more general kanban systems. Indeed, it is shown in this paper how this approach can easily be extended in order to handle kanban systems with multiple consumers and multiple suppliers, kanban-controlled assembly systems and generalized kanban systems.     

Minimising work overload in mixed-model assembly lines with different types of operators: a case study from the truck industry

This paper considers the problem of sequencing mixed-model assembly lines (MMALs). Our goal is to determine the sequence of products to minimise work overload. This problem is known as the MMAL sequencing problem with work overload minimisation: we explicitly use task operation times to find the product sequence. This paper is based on an industrial case study of a truck assembly line. In this industrial context, as a reaction to work overloads, operators at the workstations finish their tasks before the product reaches the next workstation, but at the expense of fatigue. Furthermore, there are different types of operators, each with different task responsibilities. The originality of this work is to model this new way of reacting against work overloads, to integrate three operator types in the sequencing model and to apply the developed methods in a real industrial context. To solve this problem, we propose three meta-heuristic procedures: genetic algorithm, simulated annealing and a combination of these two meta-heuristics. All the methods proposed are tested on industrial data and compared to the solutions obtained using a mixed-integer linear programme. The results show that the proposed methods considerably improve the results of the current procedure used in the case study.     

Design of a walking cyclic gait with single support phases and impacts for the locomotor system of a thirteen-link 3D biped using the parametric optimization

The development of an algorithm of parametric optimization to achieve optimal cyclic gaits in space for a thirteen-link 3D bipedal robot with twelve actuated joints is proposed. The cyclic walking gait is composed of successive single support phases and impulsive impacts with full contact between the sole of the feet and the ground. The evolution of the joints are chosen as spline functions. The parameters to define the spline functions are determined using an optimization under constraints on the dynamic balance, on the ground reactions, on the validity of impact, on the torques, and on the joints velocities. The cost functional considered is represented by the integral of the torques norm. The torques and the constraints are computed at sampling times during one step to evaluate the cost functional for a feasible walking gait. To improve the convergence of the optimization algorithm the explicit analytical gradient of the cost functional with respect to the optimization parameters is calculated using the recursive computation of torques. The algorithm is tested for a bipedal robot whose numerical walking results are presented.