Oxidative and carbonaceous patterning of Si surface in an organic media by scanning probe lithography

A simple top-down fabrication technique that involves scanning probe lithography on Si is presented. The writing procedure consists of a chemically selective patterning in mesitylene. Operating in an organic media is possible to perform local oxidation or solvent decomposition during the same pass by tuning the applied bias. The layer deposited with a positively biased tip with sub-100-nm lateral resolution consists of nanocrystalline graphite, as verified by Raman spectroscopy. The oxide pattern obtained in opposite polarization is later used as a mask for dry etching, showing a remarkable selectivity in SF₆ plasma, to produce Si nanofeatured molds.     

Prolonged display or rapid internalization of the IgG-binding protein ZZ anchored to the surface of cells using the diphtheria toxin T domain

We have shown previously that the diphtheria toxin transmembranedomain (T) may function as a membrane anchor for soluble proteinsfused at its C-terminus. Binding to membranes is triggered byacidic pH. Here, we further characterized this anchoring device.Soluble proteins may be fused at the N-terminus of the T domainor at both extremities, without modifying its membrane bindingproperties. This allows one to choose the orientation of theprotein to be attached to the membrane. Maximum binding to thecell surface is reached within 1 h. Anchoring occurs on cellspreviously treated with proteinase K, suggesting that T interactswith the lipid phase of the membrane without the help of cellsurface proteins. Binding does not permeabilize cells or affectcell viability, despite the fact that it permeabilizes liposomesand alters their structure. When attached to L929 fibroblasts,the proteins are not internalized and remain displayed at theirsurface for more than 24 h. When bound to K562 myeloid cells,the molecules are internalized and degraded. Thus, dependingon the cell type, soluble proteins may be anchored to the surfaceof cells by the T domain for an extended time or directed towardsan internalization pathway.     

Comparison of methods of differentiating between fresh and frozen–thawed fish or fillets

Freezing is an efficient way of storing fish. Objectively though, it is very hard to determine whether a fish has been previously frozen. Following an appraisal of various methods, we selected a physical determination (torrymeter), a physiological examination (eye lens) and three enzymatic assays (α‐glucosidase, β‐N‐acetylglucosaminidase and β‐hydroxyacyl‐CoA‐dehydrogenase) and applied them to three species: plaice (Pleuronectes platessa), whiting (Merlangus merlangus) and mackerel (Scomber scombrus). We also compared the results obtained following slow and rapid freezing and investigated how spoilage affects the torrymeter measurements and α‐glucosidase assay values. For whole fish the physical method using the torrymeter is a reliable indicator. For fish fillets we recommend the enzymatic method using the α‐glucosidase assay, which should be accompanied by measurement of the freshness to avoid confusing a frozen–thawed fish and a fish in an advanced stage of spoilage. The values noted for fresh and thawed whiting and plaice indicated cut‐off values of 0.15 for whiting and 0.5 for plaice, above which it can be asserted that the sample had been frozen. © 2002 Society of Chemical Industry     

Eicosapentaenoic Acid,Arachidonic Acid and Eicosanoid Metabolism in Juvenile Barramundi Lates calcarifer

A two part experiment was conducted to assess the response of barramundi (Lates calcarifer; initial weight = 10.3 ± 0.03 g; mean ± S.D.) fed one of five diets with varying eicosapentaenoic acid (diets 1, 5, 10, 15 and 20 g/kg) or one of four diets with varying arachidonic acid (1, 6, 12, 18 g/kg) against a fish oil control diet. After 6 weeks of feeding, the addition of EPA or ARA did not impact on growth performance or feed utilisation. Analysis of the whole body fatty acids showed that these reflected those of the diets. The ARA retention demonstrated an inversely related curvilinear response to either EPA or ARA. The calculated marginal utilisation efficiencies of EPA and ARA were high (62.1 and 91.9 % respectively) and a dietary ARA requirement was defined (0.012 g/kg^0.796/day). The partial cDNA sequences of genes regulating eicosanoid biosynthesis were identified in barramundi tissues, namely cyclooxygenase 1 (Lc COX1a, Lc COX1b), cyclooxygenase 2 (Lc COX2) and lipoxygenase (Lc ALOX‐5). Both Lc COX2 and Lc ALOX‐5 expression in the liver tissue were elevated in response to increasing dietary ARA, meanwhile expression levels of Lc COX2 and the mitochondrial fatty acid oxidation gene carnitine palmitoyltransferase 1 (Lc CPT1a) were elevated in the kidney. A low level of EPA increased the expression of Lc COX1b in the liver. Consideration should be given to the EPA to ARA balance for juvenile barramundi in light of nutritionally inducible nature of the cyclooxygenase and lipoxygenase enzymes.     

Polymerization shrinkage and porosity profile of dual cure dental resin cements with different adhesion to dentin mechanisms

This research aims to probe the porosity profile and polymerization shrinkage of two different dual cure resin cements with different dentin bonding systems. The self‐adhesive resin cement RelyX U200 (named RU) and the conventional Allcem Core (named AC) were analyzed by x‐ray microtomography (μCT) and Scanning Electron Microscopy (SEM). Each cement was divided into two groups (n = 5): dual‐cured (RUD and ACD) and self‐cured (RUC and ACC). μCT demonstrated that the method of polymerization does not influence the porosity profile but the polymerization shrinkage. Fewer concentration of pores was observed for the conventional resin cement (AC), independently the method used for curing the sample. In addition, SEM showed that AC has more uniform surface and smaller particle size. The method of polymerization influenced the polymerization shrinkage, since no contraction for both RUC and ACC was observed, in contrast with results from dual‐cured samples. For RUD and ACD the polymerization shrinkage was greater in the lower third of the sample and minor in the upper third. This mechanical behavior is attributed to the polymerization toward the light. µCT showed to be a reliable technique to probe porosity and contraction due to polymerization of dental cements.     

Study of ITER equatorial port plug handling system and vacuum sealing interface

In the field of the ITER port plug engineering and integration task, CEA has contributed to define proposals concerning the port plugs vacuum sealing interface with the vessel flange and the equatorial plug handling.The 2001 baseline vacuum flange sealing consisted of TIG welding of a 316L strip plate on to U shapes. This arrangement presented some issues like welding access, implementation of tools, lip consumption, complex local leak test, continuous leak checking. Therefore, an alternate sealing solution based on the use of metallic gaskets is proposed. The different technical aspects are discussed to explain how this design can simplify the maintenance and deal with safety and vacuum requirements.The design of the mechanical attachment and vacuum sealing of the plug has constantly evolved, but the associated remote handling equipment was not systematically reviewed. An update of the cask and maintenance procedure was studied in order to design it in accordance with the last generic plug flange design. This includes a concept of a gripping system that uses the plug flange bolting area and, to help the remote handling process, a cantilever assisting system is suggested to increase the reliability of the transfer operation between vacuum vessel and cask.     

Cold groundwater temperatures and conductive heat flow in the Mt. Amiata geothermal area, Tuscany, Italy

The hydrogeochemical study of the cold springs present in the Mt. Amiata geothermal area, where the Bagnore and Piancastagnaio geothermal fields are situated, has defined the different shallow groundwater systems.The cold groundwater temperature of the volcanic phreatic aquifer is largely correlated with the geothermal heat flow. Through the analysis of the temperature of cold groundwaters, a possible method for geothermal prospection has been developed, supported by the results obtained in the studied area. Through the enthalpic balance of the aquifer and in agreement with available data, a geothermal flow of about 200 mW m^-2 has been calculated.     

Biological Effects of Cyclin-Dependent Kinase Inhibitors Ribociclib,Palbociclib and Abemaciclib on Breast Cancer Bone Microenvironment

The CDK4/6 inhibitors (CDKi) palbociclib, ribociclib, and abemaciclib are currently approved in combination with anti-estrogen therapy for the treatment of advanced and/or metastatic hormone receptor-positive/HER2-neu-negative breast cancer patients. Given the high incidence of bone metastases in this population, we investigated and compared the potential effects of palbociclib, ribociclib, and abemaciclib on the breast cancer bone microenvironment. Primary osteoclasts (OCs) and osteoblasts (OBs) were obtained from human monocyte and mesenchymal stem cells, respectively. OC function was evaluated by tartrate-resistant acid phosphatase assay and real-time PCR; OB activity was assessed by an alizarin red assay. OB/breast cancer co-culture models were generated via the seeding of MCF-7 cells on a layer of OBs, and tumor cell proliferation was analyzed using flow cytometry. Here, we showed that ribociclib, palbociclib, and abemaciclib exerted similar inhibitory effects on the OC differentiation and expression of bone resorption markers without affecting OC viability. On the other hand, the three CDKi did not affect the ability of OB to produce bone matrix, even if the higher doses of palbociclib and abemaciclib reduced the OB viability. In OB/MCF-7 co-culture models, palbociclib demonstrated a lower anti-tumor effect than ribociclib and abemaciclib. Overall, our results revealed the direct effects of CDKi on the tumor bone microenvironment, highlighting differences potentially relevant for clinical practice.     

Structural Plasticity Is a Feature of Rheostat Positions in the Human Na+/Taurocholate Cotransporting Polypeptide (NTCP)

In the Na⁺/taurocholate cotransporting polypeptide (NTCP), the clinically relevant S267F polymorphism occurs at a “rheostat position”. That is, amino acid substitutions at this position (“S267X”) lead to a wide range of functional outcomes. This result was particularly striking because molecular models predicted the S267X side chains are buried, and thus, usually expected to be less tolerant of substitutions. To assess whether structural tolerance to buried substitutions is widespread in NTCP, here we used Rosetta to model all 19 potential substitutions at another 13 buried positions. Again, only subtle changes in the calculated stabilities and structures were predicted. Calculations were experimentally validated for 19 variants at codon 271 (“N271X”). Results showed near wildtype expression and rheostatic modulation of substrate transport, implicating N271 as a rheostat position. Notably, each N271X substitution showed a similar effect on the transport of three different substrates and thus did not alter substrate specificity. This differs from S267X, which altered both transport kinetics and specificity. As both transport and specificity may change during protein evolution, the recognition of such rheostat positions may be important for evolutionary studies. We further propose that the presence of rheostat positions is facilitated by local plasticity within the protein structure. Finally, we note that identifying rheostat positions may advance efforts to predict new biomedically relevant missense variants in NTCP and other membrane transport proteins.