Detection of single photoluminescent diamond nanoparticles in cells and study of the internalization pathway

Diamond nanoparticles are promising photoluminescent probes for tracking intracellular processes, due to embedded, perfectly photostable color centers. In this work, the spontaneous internalization of such nanoparticles (diameter 25 nm) in HeLa cancer cells is investigated by confocal microscopy and time-resolved techniques. Nanoparticles are observed inside the cell cytoplasm at the single-particle and single-color-center level, assessed by time-correlation intensity measurements. Improvement of the nanoparticle signal-to-noise ratio inside the cell is achieved using a pulsed-excitation laser and time-resolved detection taking advantage of the long radiative lifetime of the color-center excited state as compared to cell autofluorescence. The internalization pathways are also investigated, with endosomal marking and colocalization analyses. The low colocalization ratio observed proves that nanodiamonds are not trapped in endosomes, a promising result in prospect of drug delivery by these nanoparticles. Low cytotoxicity of these nanoparticles in this cell line is also shown.     

Common pattern of gene expression in response to hypoxia or cadmium in the gills of the European glass eel (Anguilla anguilla)

European eel (Anguilla anguilla) populations are in decline. Glass eel recruitment has fallen 10-fold since the early 1980s. Estuaries play a fundamental role in the life history of eels because glass eels must pass through them to reach freshwater ecosystems. Unfortunately, because of their geographical position at the upstream basin slopes, estuaries accumulate metals like cadmium and are important sites of hypoxia events. In this context, we studied the effect of the oxygen level on the ventilation of the glass eel. In parallel, glass eels were submitted to different dissolved cadmium concentrations (0, 2, and 10 microg L(-1)) under two oxygen levels (normoxia PO2 = 21 kPa and Hypoxia PO2 = 6 kPa). The expression level of various genes involved in the mitochondrial respiratory chain, in the cellular response to metal and oxidative stresses, was investigated. Our results showed that hypoxia enhances (1) ventilation of the postlarval stage and (2) Cd accumulation in gills only at the lowest metal water concentration tested (2 microg Cd L(-1)). At the gene level, Cd exposure mimics the effect of hypoxia since we observed a decrease in expression of genes involved in the respiratory chain and in the defense against oxidative stress.     

Interdependent Regulation of Polycystin Expression Influences Starvation-Induced Autophagy and Cell Death

Autosomal dominant polycystic kidney disease (ADPKD) is mainly caused by deficiency of polycystin-1 (PC1) or polycystin-2 (PC2). Altered autophagy has recently been implicated in ADPKD progression, but its exact regulation by PC1 and PC2 remains unclear. We therefore investigated cell death and survival during nutritional stress in mouse inner medullary collecting duct cells (mIMCDs), either wild-type (WT) or lacking PC1 (PC1KO) or PC2 (PC2KO), and human urine-derived proximal tubular epithelial cells (PTEC) from early-stage ADPKD patients with PC1 mutations versus healthy individuals. Basal autophagy was enhanced in PC1-deficient cells. Similarly, following starvation, autophagy was enhanced and cell death reduced when PC1 was reduced. Autophagy inhibition reduced cell death resistance in PC1KO mIMCDs to the WT level, implying that PC1 promotes autophagic cell survival. Although PC2 expression was increased in PC1KO mIMCDs, PC2 knockdown did not result in reduced autophagy. PC2KO mIMCDs displayed lower basal autophagy, but more autophagy and less cell death following chronic starvation. This could be reversed by overexpression of PC1 in PC2KO. Together, these findings indicate that PC1 levels are partially coupled to PC2 expression, and determine the transition from renal cell survival to death, leading to enhanced survival of ADPKD cells during nutritional stress.     

Functionalized carbon nanotubes are non-cytotoxic and preserve the functionality of primary immune cells

Carbon nanotubes are emerging as innovative tools in nanobiotechnology. However, their toxic effects on environment and health have become an issue of strong concern. In the present study, we address the impact of functionalized carbon nanotubes (f-CNTs) on cells of the immune system. We have prepared two types of f-CNTs, following the 1,3-dipolar cycloaddition reaction (f-CNTs 1 and 2) and the oxidation/amidation treatment (f-CNTs 3 and 4), respectively. We have found that both types of f-CNTs are uptaken by B and T lymphocytes as well as macrophages in vitro, without affecting cell viability. Subsequently, the functionality of the different cells was analyzed carefully. We discovered that f-CNT 1, which is highly water soluble, did not influence the functional activity of immunoregulatory cells. f-CNT 3, which instead possesses reduced solubility and forms mainly stable water suspensions, preserved lymphocytes' functionality while provoking secretion of proinflammatory cytokines by macrophages.     

Immunochemical quantification of heat denaturation of camel ( Camelus dromedarius ) whey proteins

The major whey proteins IgG, serum albumin and alpha-lactalbumin were purified from camel milk using gel permeation and ion-exchange chromatography. Specific antisera against each of them were raised and used to quantify their heat denaturation in early or mature milk over a range of 60-90 degrees C for 10-60 min using the single radial immunodiffusion technique. The heat denaturation midpoints for the mature milk heated 30 min were 67.2, 73.0 and 77.5 degrees C for IgG, albumin and alpha-lactalbumin respectively. The early milk was more sensitive to heat treatment with coagulation at low temperature and heat denaturation midpoints of 64.8, 71.6 and 72.6 degrees C respectively. This difference was related to the high IgG content of the early milk (12.6 mg/ml v. 0.5 mg/ml for the mature milk) and stresses the importance of monitoring the IgG level of milk to assess the absence of colostrum.     

Power quality improvement using DPC controlled three-phase shunt active filter

This paper presents a new control method entitled direct power control (DPC) for shunt active power filter (APF), which is applied to eliminate line current harmonics and compensate reactive power. Its main goal is to rebuild active and reactive source powers to be compared to references values using hysteresis control. The active power reference is issue of DC-side of inverter and reactive power reference is set to zero for unity power factor. The outputs of hysteresis controllers associated with a switching table, control the instantaneous active and reactive power by selecting the optimum switching state of the voltage source inverter (VSI). A theoretical analysis with a complete simulation of the system and experimental results are presented to prove the excellent performance of the proposed technique.     

Biocompatibility of heparin-grafted hemodialysis membranes: impact on monocyte chemoattractant protein-1 circulating level and oxidative status

This prospective observational study aimed at evaluating efficacy and biocompatibility performances of the new heparin-coated Evodial dialyzers with/without systemic heparin reduction. After a 4-week wash-out period with reference polysulfone F70S dialyzers, 6 hemodialysis patients were sequentially dialyzed with Evodial, F70S, and Evodial dialyzers using 30% heparin reduction, each period of treatment was 4 weeks. Removal rates (RR) (urea, creatinine, and β2-microglobulin), dialysis dose, and instantaneous clearances (urea and creatinine) were measured as well as inflammatory (C-reactive protein, fibrinogen, interleukin 6, tumor necrosis factor α, and monocyte chemoattractant protein-1) and oxidative stress (OS) (superoxide anion, homocysteine, and isoprostanes) parameters at the end of each study period. Patients treated with Evodial or F70S dialyzers for 4 weeks presented comparable dialysis efficacy parameters including urea and creatinine RR, dialysis dose and instantaneous clearances. By contrast, a significantly lower but reasonably good β2-microglobulin RR was achieved with Evodial dialyzers. Regarding biocompatibility, no significant difference was observed with inflammation and OS except for postdialysis monocyte chemoattractant protein-1 which significantly decreased with Evodial dialyzers. Thirty percent heparinization reduction with Evodial dialyzers did not induce any change in inflammation but led to an improvement in OS as demonstrated by a decrease in postdialysis superoxide production and predialysis homocysteine and isoprostane. This bioactive dialyzer together with heparin dose reduction represents a good trade-off between efficacy and biocompatibility performance (improvement in OS with a weak decrease in efficacy) and its use is encouraging for hemodialysis patients not only in reducing OS but also in improving patient comorbid conditions due to lesser heparin side effects.     

High Magnetic Field Annealing Dependent the Morphology and Microstructure of Nanocrystalline Co/Ni Bilayered Films

Co/Ni bilayered films were prepared on ITO glass by electrodeposition assisted with a magnetic field up to 0.5T aligned parallel to the electrode surface. The effect of a high magnetic field annealing up to 12T on morphology and microstructure of the post-deposited films was investigated by field emission scanning electronic microscopy (FE-SEM), X-ray diffraction (XRD) and atomic force microscopy (AFM). Grain shape and grain boundary in the Co/Ni morphology were modified dramatically when the high magnetic field was applied during the annealing process. Magnetic anisotropy appeared in the films due to the preferential orientation of fcc-CoNi alloy in comparison with a weaker orientation of hcp-Co. High magnetic field annealing favored to form a more homogeneous surface with smaller grain size and lower roughness, compared with the annealed samples obtained in the absence of magnetic field. The influencing mechanisms of high magnetic field annealing on the microstructure evolution in the Co/Ni electrodeposits are interpreted in terms of the overlapping effects: diffusion, recrystallization, grain growth and magnetic domains.     

Towards a verified compiler prototype for the synchronous language SIGNAL

SIGNAL belongs to the synchronous languages family which are widely used in the design of safety-critical real-time systems such as avionics, space systems, and nuclear power plants. This paper reports a compiler prototype for SIGNAL. Compared with the existing SIGNAL compiler, we propose a new intermediate representation (named S-CGA, a variant of clocked guarded actions), to integrate more synchronous programs into our compiler prototype in the future. The front-end of the compiler, i.e., the translation from SIGNAL to S-CGA, is presented. As well, the proof of semantics preservation is mechanized in the theorem prover Coq. Moreover, we present the back-end of the compiler, including sequential code generation and multithreaded code generation with time-predictable properties. With the rising importance of multi-core processors in safetycritical embedded systems or cyber-physical systems (CPS), there is a growing need for model-driven generation of multithreaded code and thus mapping on multi-core. We propose a time-predictable multi-core architecture model in architecture analysis and design language (AADL), and map the multi-threaded code to this model.