Lipidomic and transcriptomic analyses of Chlamydomonas reinhardtii under heat stress unveil a direct route for the conversion of membrane lipids into storage lipids.

Studying how photosynthetic cells modify membrane lipids in response to heat stress is important to understand how plants and microalgae adapt to daily fluctuations in temperature and to investigate new lipid pathways. Here, we investigate changes occurring in lipid molecular species and lipid metabolism genes during early response to heat stress in the model photosynthetic microorganism Chlamydomonas reinhardtii. Lipid molecular species analyses revealed that, after 60 min at 42 °C, a strong decrease in specific polyunsaturated membrane lipids was observed together with an increase in polyunsaturated triacylglycerols (TAGs) and diacylglycerols (DAGs). The fact that decrease in the major chloroplastic monogalactosyldiacylglycerol sn1-18:3/sn2-16:4 was mirrored by an accumulation of DAG sn1-18:3/sn2-16:4 and TAG sn1-18:3/sn2-16:4/sn3-18:3 indicated that newly accumulated TAGs were formed via direct conversion of monogalactosyldiacylglycerols to DAGs then TAGs. Lipidomic analyses showed that the third fatty acid of a TAG likely originated from a phosphatidylethanolamine or a diacylglyceryl-O-4'-(N,N,N,-trimethyl)-homoserine betaine lipid species. Candidate genes for this TAG synthesis pathway were provided through comparative transcriptomic analysis and included a phospholipase A2 homolog and the DAG acyltransferase DGTT1. This study gives insights into the molecular events underlying changes in membrane lipids during heat stress and reveals an alternative route for TAG synthesis.

3D cell culture to determine in vitro biocompatibility of bioactive glass in association with chitosan.

This study reports the in vitro biocompatibility of a composite biomaterial composed of 46S6 bioactive glass in association with chitosan (CH) by using 3D osteoblast culture of SaOS2. The 46S6 and CH composite (46S6-CH) forms small hydroxyapatite crystals on its surface after only three days immersion in the simulated body fluid. For 2D osteoblast culture, a significant increase in cell proliferation was observed after three days of contact with 46S6 or 46S6-CH-immersed media. After six days, 46S6-CH led to a significant increase in cell proliferation (128%) compared with pure 46S6 (113%) and pure CH (122%). For 3D osteoblast culture, after six days of culture, there was an increase in gene expression of markers of the early osteoblastic differentiation (RUNX2, ALP, COL1A1). Geometric structures corresponding to small apatite clusters were observed by SEM on the surface of the spheroids cultivated with 46S6 or 46S6-CH-immersed media. We showed different cellular responses depending on the 2D and 3D cell culture model. The induction of osteoblast differentiation in the 3D cell culture explained the differences of cell proliferation in contact with 46S6, CH or 46S6-CH-immersed media. This study confirmed that the 3D cell culture model is a very promising tool for in vitro biological evaluation of bone substitutes' properties.

Genome-wide transcriptome profiling of the early cadmium response of Arabidopsis roots and shoots.

Transcriptional regulation in response to cadmium treatment was investigated in both roots and leaves of Arabidopsis, using the whole genome CATMA microarray containing at least 24,576 independent probe sets. Arabidopsis plants were hydroponically treated with low (5 microM) or high (50 microM) cadmium concentrations during 2, 6, and 30 hours. At each time point, Cd level was determined using ICP-AES showing that both plant tissues are able to accumulate the heavy metal. RT-PCR of eight randomly selected genes confirmed the reliability of our microarray results. Analyses of response profiles demonstrate the existence of a regulatory network that differentially modulates gene expression in a tissue- and kinetic-specific manner in response to cadmium. One of the main response observed in roots was the induction of genes involved in sulfur assimilation-reduction and glutathione (GSH) metabolism. In addition, HPLC analysis of GSH and phytochelatin (PC) content shows a transient decrease of GSH after 2 and 6 h of metal treatment in roots correlated with an increase of PC contents. Altogether, our results suggest that to cope with cadmium, plants activate the sulfur assimilation pathway by increasing transcription of related genes to provide an enhanced supply of GSH for PC biosynthesis. Interestingly, in leaves an early induction of several genes encoding enzymes involved in the biosynthesis of phenylpropanoids was observed. Finally, our results provide new insights to understand the molecular mechanisms involved in transcriptional regulation in response to cadmium exposure in plants.

Anticonical anchoring and surface transitions in a nematic liquid crystal.

Recent works reported planar and conical azimuthally degenerated nematic anchorings. Here we predict an additional "anticonical" degenerated anchoring. Its energy presents two minima, parallel and perpendicular to the substrate plane, separated by a conical energy barrier. We realize this bistable anchoring on a grafted polymer brush and we observe temperature-driven transitions between the conical, planar, and anticonical degenerated anchorings. Under electric field we break the anticonical anchoring and switch between its bistable states.

Colloidal structures from bulk demixing in liquid crystals.

This experimental paper deals with phase separations of binary mixtures composed of a continuous liquid crystal phase and an isotropic dispersed phase. In contrast to isotropic binary mixtures, the investigated mixtures do not lead to a full phase separation but to a self-ordering of colloidal particles, as reported earlier (Loudet, J. C. et al. Nature 2000, 407, 611). We present here further aspects of such phase separations which include the kinetics of the phase separation, the origin of the formation of dislocation-like patterns, the influence of surfactants, chiral additives, and temperature on the formed colloidal structures. The present results show that (i) the dislocations in chain arrays can be seen as kinetically frozen defects, (ii) temperature can be used to control the size of the domains formed upon demixing, (iii) a slight change in surface chemistry, via the addition of surfactants, profoundly alters the formed colloidal structures, and (iv) chiral additives allow the formation of unique helical pearl chains which reflect the symmetry of the liquid crystal phase they are embedded in.

Segmental order and dynamics of polymer chains confined in block copolymer lamellar mesophases: NMR and dielectric relaxation studies.

The PDMS lamellar sublayers of a poly(styrene)-poly(dimethylsiloxane) diblock (PS-PDMS) and PS-PDMS-PS triblocks are investigated by NMR and dielectric spectroscopy. Some segments of the confined PDMS chains display anisotropic orientational fluctuations along the interfaces with the PS glassy blocks, whereas the others display fluctuations rather parallel to the lamellae normal. This coexistence results from a competitive ordering effect induced by the glassy interfaces and the chain-end anchoring junctions. The distribution of PDMS relaxation times within the sublayers is also examined: in particular, a slowing down of the segmental motions, together with a broadening of this distribution, are detected.

End-chain segment ordering in lamellar sublayers of a diblock copolymer.

The segmental ordering within a lamellar polystyrene-polydimethylsiloxane diblock (PS-PDMS) was examined by NMR. A non-uniform stretching of the PDMS chains was evidenced. Although chain segments displaying isotropic reorientational motions are present along the chains, relaxation measurements on the free PDMS extremities demonstrate that these latter are oriented and submitted to a non-zero constraint. This specific result allows to precise some features of the order distribution within the lamellae.

Synthesis of poly(styrene sulfonate) brushes.

Dense poly(styrene sulfonate sodium salt) brushes were prepared on silicone wafers using a two-step procedure: polystyrene (PS) chains, terminated by a reactive trichlorosilane group, were first covalently grafted, and then the PS brush was converted to a poly(styrene sulfonate) brush by a soft sulfonation reaction. Ellipsometry and infrared spectroscopy in ATR were used to characterize the samples and to optimize the procedure: in particular, the sulfonation was shown to be homogeneous along the chain backbone and the neutralization complete. In some cases, the polymer layer revealed to be quite fragile: the chains were pulled out of the brush. A consolidation treatment which consisted in grafting oligomers inbetween the long PS chains significantly increased the robustness of the layer. This might be relevant for industrial applications.

Evaluation of the capacity of the SCGE assay to assess the genotoxicity of biomaterials.

The comet test or SCGE assay, which is already widely used in other areas, has never been used to evaluate the mutagenic potential of medical biomaterials in the final form. The purpose of our study was thus to assess the comet test as a means of assessing the genotoxic potential of finished medical biomaterials. We used silicone elastomers with increasing concentrations of 4-nitroquinoline oxide, a genotoxic agent. Hydrogen peroxide was used as the positive control, and tissue culture polystyrene as the negative control. In our study, the comet test did not detect a significant difference in genotoxicity between the pure elastomer and the same elastomer containing 0.01 mg/ml 4-nitroquinoline oxide, but did detect a significant difference between two elastomers containing 0.01 and 0.3 mg/ml of 4-nitroquinoline oxide, respectively. Since, the surface properties of the samples were identical, only the chemical composition may have caused significant differences in mutagenicity. Whatever the cause of the genotoxicity detected by the SCGE assay, testing finished biomaterials using the comet assay makes it possible to evaluate interactions between biomaterials and living tissues that are much closer to actual application conditions.

Memory-free conic anchoring of liquid crystals on a solid substrate

We show that the anchoring memory of liquid crystals on solid substrates can be completely removed by grafting highly mobile polymer chains. Using grafted polystyrene, we obtain uniform, stable, and reproducible memory-free and conically degenerated anchoring of the nematic 5CB. The symmetry of the conic anchoring enables two different zenithal anchoring breakings, towards homeotropic and planar states. Under external torques we observe both of them, with weak thresholds.

Chain segment order in polymer thin films on a nonadsorbing surface: A NMR study

The dynamics of polydimethylsiloxane chains confined between air and a solid, wettable, and nonadsorbing surface is investigated using deuterium NMR. Chain segments in such molten films undergo uniaxial reorientations around the normal to the surfaces. The influence of the film thickness proves this dynamic uniaxiality involves rapid segment diffusion between both surfaces which impose a high degree of planar orientation. This segmental ordering is observed over a thickness range which depends on the degree of chain entanglements.

Evaluation of the in vitro biocompatibility of various elastomers.

A previous study highlighted the superior shock absorption of silicone rubbers compared to other elastomers. We evaluated and compared the in vitro biocompatibility of silicone-based rubbers and propose them as an alternative to conventional products. We used the MTT colorimetric test to assess cell viability and flow cytometry to evaluate cell proliferation. Tests were conducted at 24 and 72 h. Changes in cell morphology were evaluated by scanning electron microscopy. Positive (polyurethane) and negative (polystyrene) toxicity controls were included. The number of viable cells was significantly higher on polystyrene than on polyurethane. A decrease in the total number of cells from 24 to 72 h compared to the negative control was correlated with a lower percentage of S-phase cells. The differences in cell viability noted between the samples and the polystyrene control mainly resulted from an initial lack of adhesion, which was confirmed by scanning electron microscopy. The biocompatibility of the three silicone rubbers was comparable to the best of the three products currently being used. These results, combined with those of the previous study, indicate that silicone rubber could be considered for the manufacture of mouth guards.

Polyelectrolyte brushes: counterion distribution and complexation properties.

The structure of dense grafted polyelectrolyte layers has been studied with a combination of neutron reflectivity and infrared spectroscopy techniques. The polyelectrolyte brushes were made of poly(styrene sulfonate) neutralized by different counterions. Small counterions are distributed throughout the brush in order to ensure a highly local electroneutrality. In addition, they can be readily exchanged with other small ions. On the other hand, macromolecular counterions (as well as some proteins) are irreversibly trapped by the brush, but are located outside the grafted layer and cannot reach the surface.

Hardness and shock absorption of silicone rubber for mouth guards.

Silicone rubbers have general properties that make them suitable for the fabrication of custom-made mouth guards. This study evaluated the shock absorption properties and Shore A hardness of several silicone rubbers and derived products, compared their values with those of materials commonly used for the manufacture of mouth guards, and correlated the shock absorption and transmission abilities of these different materials with their Shore hardness. Silicone rubbers absorb shock better than the materials currently used for custom-made mouth guards. In addition, to adapt mouth guards to particular sports, the properties of the silicone rubbers can be appropriately modified by the addition of oils or glass fiber reinforcement. Statistical analysis of hardness values and transmitted forces for the 27 materials tested indicates that the maximum transmitted force increases with hardness. However, this relationship is not linear, and departure from linearity is greatest for minimal and maximal hardness values.

Subarachnoid midazolam: histologic study in rats and report of its effect on chronic pain in humans.

Subarachnoid administration via a catheter of a water-soluble benzodiazepine, midazolam, was tested in the control of cancer pain. First, the lack of its toxicity during constant subarachnoid administration (50 micrograms per day) was assessed in the rat. After 15 days of treatment, a histologic examination of the spinal cord revealed the same amount of fibrosis, infiltration, and deformation in the control group (n = 14), which had received only saline, as in the test group (n = 18), treated with subarachnoid midazolam. Therefore, the histologic changes observed in the spinal cord probably are related to the presence of the catheter. After these results, a mixture of 2 mg midazolam and a variable dose of subarachnoid morphine was injected in two patients presenting chronic neoplastic pain resistant to high doses of morphine. In these two cases, the addition of midazolam appeared to be effective in controlling intractable neoplastic pain.

Oral nociceptive activity in the rat superior colliculus.

Single units were recorded using extracellular glass microelectrodes in all laminae of the superior colliculus of the rat under halothane nitrous oxide anaesthesia. Fifty-one units were encountered which responded to a low intensity mechanical stimulus applied to a contralateral or bilateral field located in the oral sphere (intraoral 11, perioral 16), on the face (29) or on the rest of the body (21). Sixteen units responded to a jaw movement. Sixty-one cells were recorded which were preferentially (10) or only (51) activated (30) or inhibited (21) by noxious stimuli. Contralateral or bilateral mechanoreceptive fields located in intraoral (34) and perioral (35) areas were frequent. There is therefore a high incidence of the nociceptive representation of the mouth in the superior colliculus. The other functional properties of the nociceptive units were similar to those reported in other studies. From the subsequent histological localization of the recorded units, it appeared that the nociceptive projections from the intraoral and perioral regions to the superior colliculus reach the lateral part of the intermediate and deep layers of the superior colliculus.