The New Microtubule-Targeting Agent SIX2G Induces Immunogenic Cell Death in Multiple Myeloma

Microtubule-targeting agents (MTAs) are effective drugs for cancer treatment. A novel diaryl [1,2]oxazole class of compounds binding the colchicine site was synthesized as cis-restricted-combretastatin-A-4-analogue and then chemically modified to have improved solubility and a wider therapeutic index as compared to vinca alkaloids and taxanes. On these bases, a new class of tricyclic compounds, containing the [1,2]oxazole ring and an isoindole moiety, has been synthetized, among which SIX2G emerged as improved MTA. Several findings highlighted the ability of some chemotherapeutics to induce immunogenic cell death (ICD), which is defined by the cell surface translocation of Calreticulin (CALR) via dissociation of the PP1/GADD34 complex. In this regard, we computationally predicted the ability of SIX2G to induce CALR exposure by interacting with the PP1 RVxF domain. We then assessed both the potential cytotoxic and immunogenic activity of SIX2G on in vitro models of multiple myeloma (MM), which is an incurable hematological malignancy characterized by an immunosuppressive milieu. We found that the treatment with SIX2G inhibited cell viability by inducing G2/M phase cell cycle arrest and apoptosis. Moreover, we observed the increase of hallmarks of ICD such as CALR exposure, ATP release and phospho-eIF2α protein level. Through co-culture experiments with immune cells, we demonstrated the increase of (i) CD86 maturation marker on dendritic cells, (ii) CD69 activation marker on cytotoxic T cells, and (iii) phagocytosis of tumor cells following treatment with SIX2G, confirming the onset of an immunogenic cascade. In conclusion, our findings provide a framework for further development of SIX2G as a new potential anti-MM agent.     

Identification of a Maturation Plasma Cell Index through a Highly Sensitive Droplet Digital PCR Assay Gene Expression Signature Validation in Newly Diagnosed Multiple Myeloma Patients

DNA microarrays and RNA-based sequencing approaches are considered important discovery tools in clinical medicine. However, cross-platform reproducibility studies undertaken so far have highlighted that microarrays are not able to accurately measure gene expression, particularly when they are expressed at low levels. Here, we consider the employment of a digital PCR assay (ddPCR) to validate a gene signature previously identified by gene expression profile. This signature included ten Hedgehog (HH) pathways’ genes able to stratify multiple myeloma (MM) patients according to their self-renewal status. Results show that the designed assay is able to validate gene expression data, both in a retrospective as well as in a prospective cohort. In addition, the plasma cells’ differentiation status determined by ddPCR was further confirmed by other techniques, such as flow cytometry, allowing the identification of patients with immature plasma cells’ phenotype (i.e., expressing CD19+/CD81+ markers) upregulating HH genes, as compared to others, whose plasma cells lose the expression of these markers and were more differentiated. To our knowledge, this is the first technical report of gene expression data validation by ddPCR instead of classical qPCR. This approach permitted the identification of a Maturation Index through the integration of molecular and phenotypic data, able to possibly define upfront the differentiation status of MM patients that would be clinically relevant in the future.     

A DSP based FASTBUS board for data acquisition or high leveltrigger control in DELPHI electromagnetic calorimeters

A versatile FASTBUS board that is based on fast digital signal processors (DSPs) and analog-to-digital converters is described. It has been developed for the real-time acquisition and online processing of signals produced in different electromagnetic calorimeters of the DELPHI detector at the LEP (Large Electron Positron) collider. The board contains six TMS32010 DSPs, six piggyback cards for the analog-to-digital conversion, a set of 512-word FIFO memories for data exchange between contiguous DSPs and with external devices and a 16 K×16 random-access memory for data storage, accessible both to the DSPs and to FASTBUS in an asynchronous way, with override privilege granted to the DSPs     

Whiteness Constraints in a Unified Variational Framework for Image Restoration

We propose a robust variational model for the restoration of images corrupted by blur and the general class of additive white noises. The key idea behind our proposal relies on a novel hard constraint imposed on the residual of the restoration, namely we characterize a residual whiteness set to which the restored image must belong. As the feasible set is unbounded, solution existence results for the proposed variational model are given. Moreover, based on theoretical derivations as well as on Monte Carlo simulations, we provide well-founded guidelines for setting the whiteness constraint limits. The solution of the non-trivial optimization problem, due to the non-smooth non-convex proposed model, is efficiently obtained by an alternating directions method of multipliers, which in particular reduces the solution to a sequence of convex optimization subproblems. Numerical results show the potentiality of the proposed model for restoring blurred images corrupted by several kinds of additive white noises.     

Additive Manufacturing Approaches for Hydroxyapatite‐Reinforced Composites

Additive manufacturing (AM) techniques have gained interest in the tissue engineering field, thanks to their versatility and unique possibilities of producing constructs with complex macroscopic geometries and defined patterns. Recently, composite materials—namely, heterogeneous biomaterials identified as continuous phase (matrix) and reinforcement (filler)—have been proposed as inks that can be processed by AM to obtain scaffolds with improved biomimetic and bioactive properties. Significant efforts have been dedicated to hydroxyapatite (HA)‐reinforced composites, especially targeting bone tissue engineering, thanks to the chemical similarities of HA with respect to mineral components of native mineralized tissues. Herein, applications of AM techniques to process HA‐reinforced composites and biocomposites for the production of scaffolds with biological matrices, including cellular tissues, are reviewed. The primary outcomes of recent investigations in terms of morphological, structural, and in vitro and in vivo biological properties of the materials are discussed. The approaches based on the nature of the matrices employed to embed the HA reinforcements and produce the tissue substitutes are classified, and a critical discussion is provided on the presented state of the art as well as the future perspectives, to offer a comprehensive picture of the strategies investigated as well as challenges in this emerging field of materiomics.     

The effects of temperatures and volumetric expansion on the diffusion of fluids through solid polymers

This study examines moisture sorption behaviors of two glassy polymers, epoxy and vinylester, immersed in different fluids at two temperatures below the glass transition temperatures of the polymers. The main purpose of this study is to understand the effect of volume‐dependent temperatures and deformations on the diffusion process of solid polymers. Diffusivity coefficients are first determined by assuming the diffusion to follow the classical Fickian diffusion. In some cases, moisture sorption led to quite significant changes of volume, and the diffusion process cannot be well described by the Fickian diffusion. In such situation, the coupled deformation–diffusion model for linear elastic isotropic materials presented by Gurtin 1 is adopted, as a first approximation. This coupled deformation‐diffusion model reduces to a Fickian diffusion model when the coupling parameters are absent and the volume changes in the solid polymers during diffusion are negligible. A finite difference method is used in order to solve for the coupled deformation‐diffusion model. The model is used to predict the one‐dimensional moisture diffusion in thin plates and the multiaxial three‐dimensional moisture diffusion in dogbone specimens. The multiaxial diffusion in the dogbone specimens is used to validate the calibrated material parameters from the standard thin plate diffusion characterization. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 45151.     

Freeze-Drying Monitoring Using a New Process Analytical Technology: Toward a “Zero Defect” Process

This article presents a new process analytical technology to monitor a vial freeze-drying process taking into account the batch nonuniformity. It is based on a soft sensor, using a mathematical model of the process, and on the temperature measurement obtained using a new wireless device. It allows estimating the residual ice content in the monitored vials until the end of the primary drying stage and the values of the heat (and mass transfer) coefficients. Experiments carried out with aqueous solutions containing various excipients (sucrose, mannitol, polyvinylpyrrolidone) or with a pharmaceutical formulations show the effectiveness of the proposed system.