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.     

Applications of Low Field NMR Techniques in the Characterization of Oil Sand Mining,Extraction and Upgrading Processes

A number of techniques have previously been developed that use low field nuclear magnetic resonance (NMR) relaxometry for conventional and heavy oil reservoir characterization. In the current work, the adaptation of these algorithms for use in the oil sands industry is presented. NMR based methods have been developed for identification of water and bitumen content in ore and froth samples. Consistent algorithms have been used to analyze over 500 ore samples and 50 froth samples from the Athabasca oil sands in northern Alberta. Preliminary analyses are shown, with applications for in‐situ fluid determination using NMR logging tools and improved process control in oil sands processing plants.     

Obituary: Benjamin B. Wolman (1908-2000).

Memorializes Benjamin B. Wolman for his many contributions to psychology. He was proudest of a modification of psychoanalysis that he called the interactional approach. Wolman founded the International Organization for the Study of Group Tensions and the International Journal of Group Tensions. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Automatic tracking of individual fluorescence particles: application to the study of chromosome dynamics.

We present a new, robust, computational procedure for tracking fluorescent markers in time-lapse microscopy. The algorithm is optimized for finding the time-trajectory of single particles in very noisy dynamic (two- or three-dimensional) image sequences. It proceeds in three steps. First, the images are aligned to compensate for the movement of the biological structure under investigation. Second, the particle's signature is enhanced by applying a Mexican hat filter, which we show to be the optimal detector of a Gaussian-like spot in 1/omega2 noise. Finally, the optimal trajectory of the particle is extracted by applying a dynamic programming optimization procedure. We have used this software, which is implemented as a Java plug-in for the public-domain ImageJ software, to track the movement of chromosomal loci within nuclei of budding yeast cells. Besides reducing trajectory analysis time by several 100-fold, we achieve high reproducibility and accuracy of tracking. The application of the method to yeast chromatin dynamics reveals different classes of constraints on mobility of telomeres, reflecting differences in nuclear envelope association. The generic nature of the software allows application to a variety of similar biological imaging tasks that require the extraction and quantitation of a moving particle's trajectory.     

Markovian modelling and Fisher distribution for unsupervised classification of radar images

Statistical segmentation techniques based on hidden Markov field modelling have generated considerable interest in past years. They take contextual information into account in a particularly elegant and rigorous way. Although these models have been thoroughly tested, they can fail in some cases such as the non-stationary one. In this article, we propose use of the recently developed triplet Markov field, which models non-stationary images, and that of Fisher distribution, which is adapted to a wide range of surfaces for modelling synthetic aperture radar (SAR) image noise. Examples illustrate the difference between the approach proposed and classical ones. Various experiments indicate that the new model and its associated unsupervised algorithm perform better than classical ones.     

Impact of incomplete DNase I treatment on human macrophage proteome analysis

The aim of our study was to analyze the proteomic pattern of human macrophages obtained over a 4 year period from blood donors. The purpose was to simulate a long-term clinical study to assess the application of 2-D DIGE technique for differential proteomic analysis of these scarce samples. Bioinformatic analysis of 2-D DIGE gels of 19 different cultures of macrophages assessed whether they did or did not contain at least specific five spots identified by MS as being or containing bovine deoxyribonuclease I (DNase I). Bovine DNase I was used during sample treatment to remove nucleic acids from protein extracts. Macrophages were classified in two groups, which appeared to be differentiated by the completeness of DNase I treatment. Further detailed analysis revealed a different proteomic pattern of macrophage protein samples according to the completeness of this treatment. The major group of proteins affected, accounting for one third of the differentially expressed proteins, included proteins involved in cell motion and actin cytoskeleton reorganization. The use of DNase I for the removal of nucleic acids from protein samples must be avoided in proteomic studies since it can generate bias in the analysis of protein expression patterns.     

Polyphenoloxidases from Williams Pear (Pyrus communis L,cv Williams): Activation,Purification and Some Properties

Activation of a crude polyphenoloxidase (PPO) preparation extracted from Williams pears was investigated. Comparison between several activation agents led to the hypothesis of a limited conformational change involved in the activation process. Using ammonium sulphate precipitation followed by hydrophobic and ion exchange chromatography, two PPO fractions, F1 and F2, were 124- and 36-fold purified. F1 contained two forms characterised by isoelectric point equal to 4·2 and 4·5 while F2 contained two other forms associated with isoelectric point of 3·8 and 4·0. The molecular mass determined by gel filtration and confirmed after SDS-page was unique (c 43 kDa). F1 and F2 showed similar apparent Km values, in the 5–11 mM range for the three main phenolic substrates in pear cortex. Chlorogenic acid appeared to be a better substrate than catechins for pear PPO. © 1997 SCI.     

Processing, repairing and cyclic oxidation behaviour of sol–gel thermal barrier coatings

Sol–gel Thermal Barriers Coatings (TBCs) are manufactured using the dip-coating technique optimised in terms of process parameters including sol formulation, rate of withdrawing and heat treatment. The specific mechanisms of sol–gel TBCs, deposited on either NiAl or NiPtAl bond-coated superalloy substrates, are described. The possibility to reinforce and stabilise the crack network formed during the heat treatment or the first oxidation cycles using supplementary dip-coatings and appropriate process parameters is investigated. It is shown that implementing this technique that can be further regarded as an attractive way for repairing TBCs, significantly improves the cyclic oxidation behaviour of the multi-materials systems.