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Background: There is presently no cure for locomotor deficits after spinal cord injury (SCI). Very few therapies effectively target the brain due to poor understanding of the brain’s role post-SCI. Newly developed tissue clearing techniques have permitted unbiased three-dimensional circuit analysis, opening new opportunities for SCI-related brain interrogation. Methods: We established a novel brain interrogation pipeline by optimizing mouse brain clearing, imaging, and atlas registration. We leveraged a spontaneous recovery lateral hemisection model to analyze whole brain cell activity and connectivity with the lumbar cord using cFos immunolabelling and virus-mediated projection tracing. We identified a functionally and anatomically dynamic region correlating with recovery and interrogated its locomotor role with optogenetics. We assessed deep brain electrical stimulation (DBS) of this region in a more clinically relevant rat contusion SCI using an established bipedal robotic interface. Results: We unexpectedly uncovered the lateral hypothalamus (LH) to functionally and anatomically correlate with recovery. LHVglut2 optogenetic stimulation significantly augmented locomotor function. LH DBS in rats acutely robustly augmented bipedal locomotion post-SCI. Conclusions: This is the first demonstration of the LH’s role in locomotion post-SCI and is a novel DBS target that robustly augmented locomotor function, dependent on LH glutamatergic cells. LH DBS may be a promising intervention in humans.

We present the experimental realization of photonic devices optimized by a pixel-by-pixel binary optimization method, which can straightforwardly take into account the technological constraints such as minimum feature sizes in the fabrication process. In this approach, for each iteration, one considers all the candidate structures that differ by a single pixel from the starting structure and update by adopting the structure that has the best figure of merit among all candidate structures. This approach can be implemented with high computational efficiency using a Green's function method. The devices optimized by this approach have been realized on 200 mm and 300 mm Silicon-On-Insulator platforms, using either e-beam lithography or deep-UV immersion lithography. Characterizations experimentally demonstrate the reliability of the method, given the technological constraints of Silicon Photonics.

This paper deals with the improvement of the TTV values (Total Thickness Variation) of 300 mm silicon wafers thinned down at 100 μm using the ZoneBond ® (ZoneBond® is registred by Brewer Science Inc.) technology. We have demonstrated that the thin film TTV of the film was mainly driven by the bonding process and was very tolerant towards the coating step. It was also shown that, within the experiment precision, the TTV value was not changed by the grinding thinning process. 8 µm was the best TTV value obtained after the optimization of the bonding parameters.

BACKGROUND Sirtuin 1 (SIRT1) is a class III histone deacetylase that may play a critical role in several biological functions, including lifespan, stress, and inflammation. Our main objective was to evaluate SIRT1 activity in peripheral blood mononuclear cells (PBMCs) in patients with osteoporosis and to analyze the relationship between the SIRT 1 activity and markers of inflammation and bone remodelling. MATERIAL AND METHODS We performed a prospective monocentric study of patients with osteoporosis and measured the nuclear and cytoplasmic activities of SIRT1 in PBMCs. Levels of proinflammatory cytokines were assessed in culture supernatants of PBMCs isolated from the osteoporosis patients. The level of serum C-terminal cross-linking telopeptide of type I collagen (CTX), a marker of bone resorption, was measured in the serum of osteoporosis patients. RESULTS Sixteen women with osteoporosis were included. A statistically significant correlation between the cytoplasmic and nuclear SIRT 1 activities was found in PBMCs of patients with osteoporosis. Although non-significant, we observed a negative trend between nuclear SIRT 1 activity and the rate of serum CTX and a positive trend between IL-6 and CTX levels in patients with osteoporosis. CONCLUSIONS This study shows that the cytoplasmic and nuclear SIRT 1 activities are measurable in circulating PBMCs of patients with osteoporosis and that these 2 activities are correlated. The potential role of inflammation in bone resorption in patients with osteoporosis was also studied.

For the first time, a full 3D CMOS over CMOS CoolCube™ integration is demonstrated with a top level compatible with state of the art high performance FDSOI (Fully-Depleted Silicon On Insulator) process requirements such as High-k/metal gate or raised source and drain. Functional 3D inverters with either PMOS or NMOS on the top level are highlighted. Furthermore, Si layer transfer above a 28nm W Metal 1 level of an industrial short loop and the return in a front end environment is presented, confirming the industrial compatibility of CoolCube™ integration.

After a short reminder of the principle of monolithic 3D integration, this paper firstly reviews the main technological challenges associated to this integration and proposes solutions to assess them. Wafer bonding is used to have perfect crystalline quality of the top layer at the wafer scale. Thermally stabilized silicide is developed to use standard salicidation scheme in the bottom layer. Finally a fully depleted SOI low temperature process is demonstrated for top layer processing (overall temperature kept below 650^oC). In a second part the electrical results obtained within this integration scheme are summarized: mixed Ge over Si invertor is demonstrated and electrostatic coupling between top and bottom layer is used to shift the threshold voltage of the top layer. Finally circuit opportunities such as stabilized SRAM or gain in density are investigated.

The aim of this study was to assess the efficiency of aspect ratio trapping in improving the crystalline quality of relaxed Ge(1 1 0) layers selectively grown in trenches surrounded by SiO 2 . The 400 °C growth of a few hundreds of nanometers thick Ge layers has first been studied on blanket Si(1 1 0) surfaces then in recessed areas of Si(1 1 0) patterned wafers. The influence of 1 min H 2 anneals (in-between 600 and 850 °C) on the surface morphology, crystalline quality and strain state of blanket Ge(1 1 0) layers has notably been quantified. Intermediate annealing temperatures (750 °C) have improved the crystalline quality and increased the macroscopic strain relaxation of those layers, without too high a surface roughening. (1 1 0) Si windows (surrounded by shallow trench isolation) of patterned wafers have then been recessed by ≈300 nm using gaseous HCl, with a definite faceting and a slight roughening of the resulting cavities. Epitaxial lateral overgrowth of Ge has then been implemented in those trenches, followed by 1' H 2 bakes at 750 °C. Chemical mechanical polishing has been used afterwards to get rid of the several hundreds of nm thick Ge layer overflowing on the SiO 2 areas (very reduced dishing and flat Ge(1 1 0) surfaces obtained in the end). The efficiency of aspect ratio trapping in reducing the defect density in those Ge(1 1 0) layers is not obvious. Indeed, some trapping of inclined defects in the SiO 2 sidewalls of narrow Ge(1 1 0) patterns (∼80 nm long) has been evidenced. However, the theoretically unexpected appearance of defects at 90° to the surface (i.e. normal to (1 1 0)) that were consequently not trapped was detrimental to defect density reduction. Those 90° defects may have arisen from interactions of inclined defects with one another. The reduction of the high defect density in relaxed (1 1 0) layers is thus still challenging and requires further investigations.

This paper is dedicated to the full integration of innovative silicon-based material for Wafer-Level molding of silicon interposer wafers. This technology can be used in the frame of silicon packages where the silicon interposer is either reported on P-BGA or directly assembled on board. Interest of such material, is the rapid Wafer-Level lamination process and die planarization which can facilitate interposer realization. In a first part of the article, we have evaluated the compatibility of this material with the whole interposer flow by having a focus on filling capacities as well as induced deformation on 300mm wafers. Secondly, we have generated electrical test vehicles in order to verify the impact of such material on environmental and mechanical reliability.

We have studied at 20 Torr the very high temperature (i.e.850ºC to 1100ºC) growth kinetics of SiGe in Reduced Pressure-Chemical Vapour Deposition. The growth chemistry adopted was heavily chlorinated, i.e. SiH2Cl2 + GeH4 + HCl, in order to avoid any significant chamber quartz wall coating (and thus flaking) during the deposition of thick layers. We are mostly in a high temperature, supply limited regime, with a small increase of the SiGe growth rate as the GeH4 mass flow and/or the growth temperature increases. By contrast, the Ge concentration decreases for given precursor mass flows as the growth temperature increases. Its dependence on the F(GeH4)/F(SiH2Cl2)+F (GeH4) mass flow ratio, almost linear at 850ºC and 900ºC, gets parabolic for higher growth temperatures. We have used those datapoints to grow linearly graded (~9%Ge/m) SiGe Virtual Substrates(VS) with final Ge concentrations in the 15% to 45% range.

Resume L'inflammation est une reaction le plus souvent transitoire parce qu'elle est rapidement interrompue par des mecanismes inhibiteurs. Le premier de ces mecanismes est le blocage des voies de signalisation associees aux recepteurs des mediateurs de l'inflammation. Il implique que des facteurs de transcription responsables du developpement de l'inflammation soient inactives par des proteines presentes constitutivement ou exprimees de novo sous le controle des mediateurs de l'inflammation eux-memes. Le deuxieme mecanisme est la degradation ou l'inactivation des mediateurs de l'inflammation. Le dernier mecanisme est l'arret de la synthese des mediateurs de l'inflammation. Il apparait en particulier que plusieurs precurseurs de ces mediateurs sont progressivement detournes d'une voie metabolique favorisant la formation de mediateurs pro-inflammatoires au profit d'une autre voie a l'origine de mediateurs anti-inflammatoires. Tous ces mecanismes sont la cible de nouvelles approches therapeutiques.

In this letter, we report the fabrication and characterization of self-aligned double-gate MOSFETs with gate length down to 6 nm. Based on molecular bonding, the interest of this original process relies on the fact that, for the first time, technological options such as planar process, independently biasable gates, and metallic source and drain are integrated all together to address critical issues for sub-22-nm node, such as variability, short channel effect control, and access resistance decrease. Good electrical performance of pMOS transistors is demonstrated. Short channel effects are very well controlled down to 30 nm. The independent biasing of the two gates allows tuning of the characteristics, depending on the targeted applications.

The reactions induced in a vacuum furnace (5×10−7 Torr) between an electron-beam-evaporated Ni film a few hundred nm thick and a (001)-oriented (i.e. Si-face-oriented) single crystalline 3C–SiC substrate are investigated by 3.2 MeV 4He2+ backscattering spectrometry, X-ray diffraction, secondary ion mass spectrometry, and scanning electron microscopy. Samples are characterized before and after annealing at temperatures of 400–700oC for 30 min. At 450oC, carbon diffuses throughout the Ni film and forms a carbon-rich layer at the Ni surface of a thickness of a few nm which remains unchanged during subsequent annealing. Some nickel silicides were detected at this initial stage but could not be clearly identified. At 450oC (after 120 min) the Ni31Si12 phase starts to form. This is the only detected phase at 500oC. The Ni2Si phase, the silicide that is thermodynamically stable with SiC and carbon, forms first at the surface and grows toward the SiC substrate. At 600oC, this reaction has consumed about half of the Ni31Si12 phase and at 700oC, Ni2Si is the only silicide in the reacted film. In all the reacted samples the carbon distribution is alike and consists of three distinct layers: a first zone with a constant carbon concentration that extends from near the SiC/silicide interface through most of the films thickness. The second zone is ∼70 nm thick and is deficient of carbon. The third zone is the thin graphite layer at the surface. There is oxygen in the film too, the distribution of which is related increasingly clearly to the carbon profile as the annealing temperature rises.

Stable fluorine doped PETEOS and HDP-CVD silicon oxide thin films have been deposited without any capping layer. Gap-filling of 0.35 /spl mu/m metal spacing (2:1 A.R.) has been studied, and partial integration has been successfully achieved.

Macrophage arginine metabolism via nitric oxide (NO) synthase and arginase pathways reduces and enhances tumor cell proliferation, respectively. Transforming growth factor-beta (TGF-beta) has been shown to down-regulate the NO synthase pathway. The present study describes the effect of TGF-beta on the arginase pathway. TGF-beta up-regulated arginase activity in rat peritoneal macrophages as assessed by measuring the generation of [14C]urea from [14C]-L-arginine in the presence of NG-monomethyl-L-arginine (L-NMMA). The stimulation, which reached fivefold after a 48-h exposure of macrophages to 10 ng/ml TGF-beta, was due to reduction in Km value of arginase. TGF-beta-induced up-regulation of arginase activity led to the release of more polyamines, mainly putrescine. The role of this up-regulation on macrophage cytotoxicity toward L-929 tumor cells was analyzed in coculture experiments. Macrophages blunted DNA synthesis by L-929 cells as assessed by measuring the incorporation of [3H]TdR into the cells and the proportion of cells in the G2 phase. Addition of TGF-beta in the presence of L-NMMA permitted L-929 cells cocultured with macrophages to resume DNA synthesis. The mechanism responsible for this restoration was the up-regulation of arginase activity rather than the down-regulation of NO synthase activity since TGF-beta in the presence of L-NMMA failed to further reduce NO synthase activity whereas it still enhanced arginase activity; synthetic putrescine (1-10 microM) also blunted macrophage cytotoxicity toward L-929 cells. This is the first evidence that TGF-beta up-regulates arginase activity in macrophages and, hence, limits macrophage-dependent cytostasis.

In this work, both surface and bulk interactions between Fe and SiC are studied. Surface interactions are quantified by contact angles measured by the sessile drop technique under high vacuum. Products of Fe-SiC bulk reactions are characterized by scanning electron microscopy and microprobe analysis. The interpretation of the experimental results on reactivity is based on classical thermodynamics applied to equilibria of bulk phases. The results are used to discuss the general relationship between bulk reactivity and wettability.

Interfacial reactions between a W thin film and a single-crystal (001) β-SiC substrate on rapid thermal annealing from 600 °C to 1100 °C for 60 s were investigated by backscattering spectrometry, X-ray diffraction, secondary ion mass spectrometry and cross-sectional transmission electron microscopy. Backscattering spectrometry shows that W reacts with SiC at 950 °C. The product phases identified by X-ray diffraction are W5Si3 and W2C. At 1100 °C no more unreacted W is detected. Current-voltage measurements show that ohmic contacts are already obtained on as-deposited W. Contact resistivity measured using the circular transmission line model is about 10−3 Ω cm2. Thermodynamic studies of the solid phase stability in the ternary WSiC system help us to understand the chemical stability of W thin film.

Contact resistivities of as-deposited and annealed Pt, Re and Ta films on n-type single-crystalline β-SiC(001) are characterized using a circular contact pattern and the circular transmission-line model method. The β-SiC substrates used in the experiment are n-type doped either non-intentionally to a carrier concentration of about 10 17 cm −3 , or by nitrogen implantation and annealing to a concentration of 5 × 10 19 cm −3 . The effect of a finite resistance along the circular contact rings on the measured potentials is corrected with a resistance network model. On the non-intentionally doped β-SiC substrates, Pt contacts are non-ohmic regardless of the heat treatment. The as-deposited Ta and Re contacts are ohmic with contact resistivities of 5 × 10 −5 Ω cm 2 and 4 × 10 −4 Ω cm 2 respectively. Upon annealing at 500 °C for 30 min, the resistivity of Ta increases slightly while that of Re decreases slightly. Both Ta and Re contacts become non-ohmic by annealing at 900 °C for 30 min. The as-deposited Ta, Pt and Re contacts are all ohmic on the nitrogen-implanted β-SiC substrate. The contact resistivity of the as-deposited Ta contact is the lowest and in the order of high 10 −7 Ω cm 2 , stays about the same at 500 °C and degrades to 4.3 × 10 −6 Ω cm 2 at 1000 °C. The as-deposited Re contact has the highest contact resistivity of 1 × 10 −4 Ω cm 2 but it improves to 1 × 10 −5 Ω cm 2 upon annealing at 900 °C. The contact resistivity of the as-deposited Pt contacts is 6 × 10 −6 Ω cm 2 and increases to 1 × 10 −5 Ω cm 2 at 500 °C. After annealing at 900 °C for 30 min, the Pt contact on the nitrogen-implanted β-SiC is no longer ohmic. The results are compared with the reactions that take place in those systems.

The reaction between a sputtered-deposited Ta film (320 nm thick) and a single crystalline (001) beta-SiC substrate induced by vacuum annealing at temperatures of 600-1200 C for 1 h (30 min at 1100 C) is investigated by 3 MeV He(+2) backscattering spectrometry, x-ray diffraction, secondary ion mass spectrometry, and transmission and scanning electron microscopies. No significant reaction is observed at 800 C or at lower tempertures. At 900 C, the main product phases are Ta2C and carbon-stabilized Ta5Si3. A minor amount of unreacted Ta is also present. After annealing at 1000 C, all the tantalum has reacted; the reaction zone possesses a multilayered structure of beta-SiC/TaC/carbon-stabilized Ta5Si3/alpha-Ta5Si3/Ta2C. The diffusion path at 1000 C is plotted on the isothermal section of the Ta-Si-C phase diagram. At 1100 C, the reacted layer has an interface with the SiC substrate that is still quite flat but has a rough surface due to the formation of macroscopic voids within the reacted layer. The equilibrium products predicted by the phase diagram are TaC and TaSi2. This final state is reached by annealing at 1200 C for 1 h. At that point, the reacted layer has a latterally very uneven structure and morphology.

Thermally induced solid-state reactions between a 70 nm Pt film and a single-crystal (001) β-SiC substrate at temperatures from 300 °C to 1000 °C for various time durations are investigated by 2 MeV He backscattering spectrometry, x-ray diffraction, secondary ion mass spectrometry, scanning electron microscopy, and cross-sectional transmission electron microscopy. Backscattering spectrometry shows that Pt reacts with SiC at 500 °C. The product phase identified by x-ray diffraction is Pt3Si. At 600–900 °C, the main reaction product is Pt2Si, but the depth distribution of the Pt atoms changes with annealing temperature. When the sample is annealed at 1000 °C, the surface morphology deteriorates with the formation of some dendrite-like hillocks; both Pt2Si and PtSi are detected by x-ray diffraction. Samples annealed at 500–900 °C have a double-layer structure with a silicide surface layer and a carbon-silicide mixed layer below in contact with the substrate. The SiC—Pt interaction is resolved at an atomic scale with high-resolution electron microscopy. It is found that the grains of the sputtered Pt film first align themselves preferentially along an orientation of {111}Pt//{001}SiC without reaction between Pt and SiC. A thin amorphous interlayer then forms at 400 °C. At 450 °C, a new crystalline phase nucleates discretely at the Pt-interlayer interface and projects into or across the amorphous interlayer toward the SiC, while the undisturbed amorphous interlayer between the newly formed crystallites maintains its thickness. These nuclei grow extensively down into the substrate region at 500 °C, and the rest of the Pt film is converted to Pt3Si. Comparison between the thermal reaction of SiC-Pt and that of Si–Pt is discussed.

Thermal reactions of Re thin films in contact with single crystalline (001) β‐SiC at temperatures between 700 and 1100 °C for 30 min are investigated by MeV He++ backscattering spectrometry, x‐ray diffraction, secondary ion mass spectrometry, and transmission electron microscopy (plan‐view and cross‐sectional). No reaction between Re and SiC is observed for any annealing conditions. The average grain size of the as‐deposited Re film is 220 nm and increases to 280 nm after annealing at 1100 °C for 30 min. A strong {0001}Re fiber texture is also observed after annealing. The chemical stability of Re thin films on SiC is consistent with the earlier study of solid‐phase stability in the ternary Re‐Si‐C system which shows that Re and its silicides have tie lines with SiC at 1600 °C. It also coincides with calculations of the free energy of reaction from assessed thermodynamic data for rhenium silicides and SiC. The implications of this Re stability with SiC for applications of Re as a metal for electrical cont...

3D sequential integration enables the full use of the third dimension thanks to its high alignment performance. In this paper, we address the major challenges of 3D sequential integration: in particular, the control of molecular bonding allows us to obtain pristine quality top active layer. With the help of Solid Phase Epitaxy, we can match the performance of top FET, processed at low temperature (600°C), with the bottom FET devices. Finally, the development of a stable salicide enables to retain bottom performance after top FET processing. Overcoming these major technological issues offers a wide range of applications.

3D monolithic integration, thanks to its high vertical density of interconnections, is the only available option for applications requiring connections at the transistor scale. However to achieve 3D monolithic integration, some issues such as realization of high quality top film, high stability bottom FET, low thermal budget top FET still have to be solved. In this work, a 3D monolithic process flow relying on molecular wafer bonding is proposed and results in all critical steps are given. Significant breakthroughs have been obtained using a full wafer molecular bonding with thin interlayer dielectric and an original salicidation process stabilized up to 650°C enabling to reach high performance for the top and bottom transistor. With such technology, we demonstrate functional top and bottom transistors as well as 3D structures such as invertors and SRAMs.

There is considerable evidence suggesting that reactive oxygen species (ROS) are implicated in the pathogenesis of ischemic, toxic, and immunologically-mediated renal injury. In experimental renal ischemia, ROS sources include the electron transport chain, oxidant enzymes (xanthine oxidase), phagocytes, and auto-oxidation of epinephrine. ROS cause lipid peroxidation of cell and organelle membranes and, hence, disruption of the structural integrity and capacity for cell transport and energy production, especially in the proximal tubule segment. In experimental immune glomerulonephritis, ROS are generated by both infiltrating blood-borne cells (polymorphonuclear leukocytes and monocytes) and resident glomerular cells, mainly mesangial cells. Their formation results in morphologic lesions and in modifications of glomerular permeability to proteins through activation of proteases and reduction of proteoglycan synthesis. Additionally, they promote a reduction in glomerular blood flow and glomerular filtration rate through liberation of vasoconstrictory bioactive lipids (prostaglandins, thromboxane, and platelet activating factor) and, possibly, inactivation of relaxing nitric oxide. Further studies are needed to address the role of ROS in human glomerular diseases.

We present the first experimental study of a Gate-All-Around (GAA) SONOS memory architecture with 4-level crystalline nanowire channels (down to 6nm-diameter). The technology is also extended to an independent double gate memory architecture, called φ-Flash. The experimental results with 6nm nanowires show high programming windows (up to 7.4V), making the structure compatible with multilevel operation. Excellent retention even after 104 cycles is achieved. The independent double gate option has otherwise been successfully integrated with 4-level stacked nanowires for multibit applications. The φ-Flash exhibits up to 1.8V ΔV Th between its two gates, demonstrating multibits operation. The basic process to fully disconnect the different nanowires in view of a full 3D integration of a memory array is discussed.

For the first time 3D sequential CMOS integration turns up to be an actual competitor for sub 22nm technology nodes. Thanks to the original use of molecular bonding, high quality top Si active layers are obtained. Thermally robust bottom salicide goes through the whole top FET processing without any significant sheet resistance degradation. The low temperature integration of raised source and drain for top layers is demonstrated. A decrease by 4A of the Equivalent Oxide Thickness is measured when a low thermal budget process is implemented. The electrostatic coupling between stacked FETs is demonstrated thanks to an ultra thin inter layer dielectric thickness of 60nm. It leads to a threshold voltage dynamic shift of 130mV enabling SRAM stabilization.

We report in this paper the fabrirication and the characterirization of FDSOI pMOSFETs with metallic source and drain exhibiting the best performance obtained so far on metallic source/drain devices, with Ion=345µA/µm and Ioff=30nA/µm at −1V for a 50nm gate length device. These results have been achieved thanks to a careful optimization of the source/drain to channel contacts, which can allow specifific contact resistivities as low as 0.1 Ω.µm2.

For the first time, we report a 3D stacked sub-15 nm diameter NanoWire FinFET-like CMOS technology (3D-NWFET) with a new optional independent gate nanowire structure named PhiFET. Extremely high driving currents for 3D-NWFET (6.5 mA/mum for NMOS and 3.3 mA/mum for PMOS) are demonstrated thanks to the 3D configuration using a high-k/metal gate stack. Co-processed reference FinFETs with fin widths down to 6 nm are achieved with record aspect ratios of 23. We show experimentally that the 3D-NWFET, compared to a co-processed FinFET, relaxes by a factor of 2.5 the channel width requirement for a targeted DIBL and improves transport properties. PhiFET exhibits significant performance boosts compared to Independent-Gate FinFET (IG-FinFET): a 2-decade smaller IOFF current and a lower subthreshold slope (82 mV/dec. instead of 95 mV/dec.). This highlights the better scalability of 3D-NWFET and PhiFET compared to FinFET and IG-FinFET, respectively.

Building systems that are guaranteed to be secure or to remain secure over time is still an unachievable goal. The need for a security cockpit that helps to determine security assurance level of a system in a near real time manner is therefore vital in order to maintain and improve overall security. This paper discusses different steps in the whole security assurance assessment process in order to build such cockpit. To test and evaluate the applicability of our approach, we introduce then a system capable of assessing the security assurance level of a network system. Currently, this assessment system focuses on the availability aspect of the security.

Building systems that are guaranteed to be secure or to remain secure over time is still an unachievable goal. The need for a tool that helps to determine security assurance level of a system is therefore vital in order to maintain and improve overall security. This paper introduces our system to assess the overall security assurance of a large, networked, IT-driven system in terms of a dedicated evaluation infrastructure based on multi-agent technology. We use attack graph approach to compute an attackability metric value and define other metrics for anomaly detection to assess both the static and dynamic visions of the system under study. The implemented software system is described, and the examples of experiments for evaluating of network component, sub network and network security assurance levels are considered.

We introduce a new overlay network named ROSA1. Overlay networks offer a way to bypass the routing constraints of the underlying network. ROSA used this overlay network property to offer a resilient routing to critical applications. Unlike other overlay networks dealing with the routing resilience issue, we oriented our research towards building a robust overlay network topology instead of a robust routing function. We tried to maintain a path between any pairs of nodes of the network. The routing resilience is obtained by forcing nodes to choose and modify dynamically their neighbors set according to the ROSA protocol. Moreover, ROSA is highly scalable.

The regulatory effect of H2O2 on both the cytotoxic activity and the specific binding of TNF-alpha was studied by using TNF-alpha-sensitized murine L929 cells. When these cells were exposed simultaneously to TNF-alpha and H2O2 (100 to 500 microM), the cytotoxic activity of TNF-alpha was inhibited by up to 66.6%. This inhibition was also effective when the cells were pretreated by H2O2, but not when TNF-alpha alone was preexposed to H2O2. These data suggest that H2O2 altered the cell sensitivity to TNF-alpha, without modifying the activity of the TNF-alpha molecule. Maximum loss of cell sensitivity to TNF-alpha occurred after 30-min preexposure to 500 microM H2O2. Complete restoration of TNF-alpha sensitivity was obtained within 12 h after H2O2 removal. It required protein synthesis as demonstrated by the suppressive effect of actinomycin D. The inhibitory effect of H2O2 was suppressed by catalase, but was unaffected by the scavengers of hydroxyl radical and hypochlorous acid, suggesting that H2O2 but not one of its metabolites was responsible for this inhibition. H2O2 inhibitory effect did not implicate any change in prostaglandin production or in PKC activity. In contrast, H2O2 effect was associated with an about 50% loss of the density of cell membrane 125I-TNF-alpha receptors (2949 vs 5620 binding sites per cell), without change in their affinity (3.9 vs 3.4 nM). Moreover H2O2 did not affect the rate of degradation of TNF-alpha, and only slightly increased the degree of internalization of 125I-TNF-alpha receptors. These findings indicate that H2O2 can down-regulate the cellular response to TNF-alpha, possibly by reducing the TNF-alpha-binding capacity.

This paper describes a WCDMA direct conversion receiver which has been integrated in a BiCMOS SiGe-carbon process featuring 0.25 /spl mu/m/f/sub T/=60 GHz bipolar transistors. This receiver includes an integrated RF-front-end with local oscillator quadrature generator, 5/sup th/ order Butterworth analog baseband lowpass filter (LPF) and variable gain amplifier (VGA), cut-off frequency calibrator, DAC for DC-offset calibration, serial bus interface and voltage and current reference generators. In the high/low gain modes, this device consumes 25 mA and 20 mA respectively with 2.7 V power supply. The die is wire bonded directly on the validation board. Within the receive band, the measurements show 51 dB of overall gain, NF=5 dB, IIP3= -9 dBm, ICP1 = -15dBm.

Background Sirtuin 1 (Sirt1) is a nuclear enzyme from the class III histone deacetylases (HDACs) modulating gene expression, involved in the regulation of various biological processes (cell survival, apoptosis, gluconeogenesis, adipogenesis, lipolysis), local and systemic inflammation, as well as in bone and cartilage remodeling. Sirt-1 reduces pro inflammatory cytokines release. Objectives The main objective of the study was to evaluate Sirt1 activity in PBMC by venous blood aspiration in rheumatoid arthritis (RA) patients, compared to those of control patients, and to analyze the relationship between Sirt1 activity, disease activity and production of mediators of inflammation and cytokines (TNFalpha, IL-6, IL-8) by the cells, before and after ex vivo treatment with a Sirtuin activator, resveratrol. Methods A prospective and comparative monocentric study was performed in order to compare the activity of Sirt1 in patients with RA (according to ACR criteria) and controls. Disease activity was assessed by DAS28 (ESR) and CRP. PBMC were isolated from venous blood, and Sirt1 activity was evaluated from cytoplasmic and nuclear compartments using a fluorometric assay (SIRT1 fluorimetric kit, BML-AK-555, Enzo Life Sciences, Villeurbanne, France) at the 15 minutes point. Culture supernatant levels of TNF alpha, IL-6, IL-8 were quantified before and after resveratrol (1 μmol and 5 μmol) ex vivo treatment, with commercial kits (Quantikine Kits, RD significance: p less than 0.05. Results Twenty two patients with RA (age 55±12 years, meanDAS28: 4.27;disease duration 15±10 years; 75% ACPA +; mean CRP: 12.7 mg/l) and 18 controls (age 54±13 years) were included. No differences were found in cytoplasmic or nuclear Sirt1 activity between patients and controls. Cytoplasmic and nuclear Sirt1 activity were correlated in patients and controls. Sirt1 activity (nuclear and cytoplasmic) was not correlated with DAS28, CRP or ESR, but cytoplasmic Sirt-1 activity was correlated to baseline IL-6 (p=0.02) and baseline TNF (p=0.04) in RA and controls, but not with IL-8. Sirt1 activity (cytoplasmic and nuclear) was lower in RA ACPA+ compared to ACPA neg, and nuclear activity was higher in patients treated with corticosteroids. After resveratrol treatment, no changes in TNF, IL-6 or IL-8 levels were found, Conclusions Sirt1 activity (cytoplasmic and nuclear) from PBMC was not different between RA patients and controls, nevertheless, the correlation in RA with Sirt1 activity and TNF and IL-6 levels, as well as reduced Sirt1 activity in ACPA + patients suggest an implication of this epigenetic regulation in RA. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.1584

Background Sirtuin 1 (Sirt1) is a nuclear enzyme from the class III histone deacetylases (HDACs) modulating gene expression, involved in the regulation of various biological processes (cell survival, apoptosis, gluconeogenesis, adipogenesis, lipolysis), local and systemic inflammation, as well as in bone and cartilage remodeling. Sirt-1 reduces pro inflammatory cytokines release. Spondyloarthritis (SpA) is a multifactorial disease, but no epigenetic data are currently available in this disease. Objectives The main objective of the study was to evaluate Sirt1 activity in PBMC by venous blood aspiration in spondyloarthritis (SpA) patients, compared to those of control patients, and to analyze the relationship between Sirt1 activity, disease activity and production of mediators of inflammation and cytokines (TNFalpha, IL-6, IL-8) by the cells, before and after ex vivo treatment with a Sirtuin activator, resveratrol. Methods A prospective and comparative monocentric study was performed in order to compare the activity of Sirt1 in patients with SpA (according to ASAS criteria) and controls. Disease activity was assessed by BASDAI, ASDAS, ESR and CRP. PBMC were isolated from venous blood, and Sirt1 activity was evaluated from cytoplasmic and nuclear compartments using a fluorometric assay (SIRT1 fluorimetric kit, BML-AK-555, Enzo Life Sciences, Villeurbanne, France) at the 15 minutes point. Culture supernatant levels of TNF alpha, IL-6, IL-8 were quantified before and after resveratrol (1 μmol and 5 μmol) ex vivo treatment, with commercial kits (Quantikine Kits, RD significance: p less than 0.05. Results Forteen patients with SpA (age 48±14 years, mean BASDAI 43±18; ASDAS-CRP 3,1±0,9; disease duration 19±10 years; all were HLA-B27+; mean CRP: 19.1 mg/l) and 18 controls (age 54±13 years) were included. No differences were found in cytoplasmic or nuclear Sirt1 activity between patients and controls. Cytoplasmic and nuclear Sirt1 activity were correlated each other in patients and controls. Sirt1 activity (nuclear and cytoplasmic) was not correlated with ASDAS, BASFI, CRP or ESR, but a trend of correlation was found between nuclear Sirt-1 activity and BASDAI (r =0.5; p=0.07). No differences in Sirt1 activity was found between axial and peripheral forms of SpA. Sirt1 activity (cytoplasmic and nuclear) was higher in SpA with NSAIDs compared to patients without NSAIDs (p=0.005). No correlations were found between Sirt1 activity and cytokine levels at baseline, and no change after ex vivo treatment by resveratrol. Conclusions Sirt1 activity (cytoplasmic and nuclear) from PBMC was not different between SpA patients and controls in this study, these result do not argue for a major implication of Sirt1 in SpA. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.1744

Transforming growth factor-beta (TGF-beta1) enhances interleukin-10 (IL-10) synthesis by mouse monocytes/macrophages, suggesting a potential role of IL-10 in mediating some of the anti-inflammatory properties of TGF-beta1. Since differences exist between the transcriptional regulation of human and mouse IL-10, the studies reported here examined whether TGF-beta1 up-regulated IL-10 production by human monocytes/macrophages as well. Exposure of PMA-differentiated U-937 promonocytic cells to TGF-beta1 resulted in an unexpected, dose-dependent decrease in IL-10 production as assessed by specific ELISA. TGF-beta1 was effective when added at the time of the PMA stimulus or 6 hours after. In addition, TGF-beta1 suppressed induction of IL-10 by three different stimuli other than PMA. TGF-beta1 inhibition of IL-10 protein release was associated with proportional changes in IL-10 mRNA accumulation as assessed by quantitative kinetic ELISA PCR. This would result from a decrease in IL-10 gene transcription as TGF-beta1 did not affect IL-10 mRNA stability, and TGF-beta1 limited the luciferase activity in cells transfected with reporter gene constructs containing 1,308 bp of the 5' non-coding sequence of human IL-10 gene. Blocking tumour necrosis factor-alpha (TNF-alpha) with neutralizing anti-TNF-alpha antibody did not modify the response to TGF-beta1, indicating the involvement of TNF-alpha-independent mechanisms in the overall process. Thus, the present study provides the first evidence that TGF-beta1 prevents IL-10 production by human monocytic cells at a transcriptional level.

Both pro- and anti-inflammatory mediators regulate the anti-inflammatory actions of glucocorticoids, in part by modifying the binding of glucocorticoids to specific receptors. For instance, somatostatin has been shown to increase glucocorticoid binding and signaling in macrophages. The mechanism of this regulation does not require an increased expression of glucocorticoid receptors but, rather, a stabilization of glucocorticoid receptor-associated heat shock protein 90. This is related to a decrease in calpain activity. Thus calpain inhibition may offer a new and exciting possibility for enhancing the anti-inflammatory efficiency of glucocorticoids.

Studies of glomerulonephritis models have shown that inflammatory reaction is responsible for the development of glomerulosclerosis and tubulo-interstitial sclerosis and, hence, for the progression to end stage renal failure. That macrophage accumulation and fibrosis extension are frequently not closely related events suggests that macrophages are not involved in progression process. Glomerular sclerosis is rather associated with the release of mediators from resident cells-mainly growth factors such as platelet-derived growth factor and transforming growth factor-beta--the synthesis and bioactivity of which are enhanced by inflammatory mediators. Tubulo-interstitial sclerosis is induced by inflammatory lesions of the glomerulus that lead to proteinuria. Indeed, reabsorption of proteins in proximal tubule triggers epithelial cells to release proinflammatory and prosclerotic mediators into the interstitium. New therapeutic approaches including gene transfer strategies are directed at suppressing the efficiency of such mediators.

Intracellular dehydration is the major risk in case of a polyuropolydipsic syndrome. Excepting osmotic polyuria, prognosis depends on a possibly progressive functional anomaly of the hypothalamopituitary axis.Polyuropolydipsia occurs when antidiuretic hormone (ADH) secretion is absent (central diabetes insipidis), the kidney does not respond to ADH (nephrogenic diabetes insipidus) or in case of physiological inhibition of ADH secretion (primary polydipsia).Dynamic explorations are associated with radioimmunoassay of ADH. They are particularly useful in case of atypical diabetes insipidus and include the water restriction test and a study of the sensitivity to exogenous ADH (dDAVP). The results orient the etiologic diagnosis and allow an evaluation of the fluid intake required as a function of the maximal concentrating capacity of the kidneys.Treatment is based on ADH analogs (dDAVP). The aim is to obtain a constant antidiuretic effect without hyponatremia or escape. In case of partial central diabetes insipidus, a non-hormone treatment using compounds which increase vasopressin release or its effect on the kidney can be proposed.

Stimulation of macrophages with endotoxin and/or cytokines is responsible for the expression of the inducible isoform of nitric oxide synthase (iNOS). Because macrophages are exposed to low pH within the microenvironment of inflammatory lesions, the potential role of acidic pH as an additional regulator of iNOS was investigated. Substitution of the culture medium of rat peritoneal macrophages at pH 7.4 with medium at pH 7.0 up-regulated iNOS activity, as reflected by a 2.5-fold increase in nitrite accumulation. The increase in iNOS activity was associated with a similar increase in iNOS mRNA expression that reflected an increase in iNOS mRNA synthesis rather than stability. Low environmental pH-induced iNOS gene transcription involved the activation of nuclear factor-kappaB (NF-kappaB) transcription factor since exposure of macrophages to low environmental pH both increased NF-kappaB binding activity in the nucleus and enhanced NF-kappaB-driven reporter gene expression. In addition, treatment of macrophages with pyrrolidine dithiocarbamate or n-acetyl-leucinyl-leucinyl-norleucinal, two drugs preventing NF-kappaB translocation to the nucleus, canceled low pH-induced nitrite accumulation. The overall mechanism required the synthesis of tumor necrosis factor alpha (TNFalpha). Indeed, 1) elevated TNFalpha bioactivity was observed in the medium of macrophages exposed to pH 7.0, and 2) incubation of macrophages with a neutralizing anti-TNFalpha antibody impaired both NF-kappaB activation and nitrite accumulation in response to acid challenge. In summary, exposure of macrophages to acidic microenvironment in inflammatory lesions leads to the up-regulation of iNOS activity through the activation of NF-kappaB.

We investigated the expression and potential regulatory role of insulin-like growth factors (IGFs) and their specific binding proteins (BPs) in tuberculous and nontuberculous pleuritis. By using a radioimmunoassay after acid gel filtration chromatography, we found that mean concentrations of IGF-I were 211.9 +/- 20.2 microg/l and 203.2 +/- 31.1 microg/l in pleural fluid of 14 patients with tuberculous pleuritis and 9 patients with malignant pleuritis respectively. These values were near those in serum of the same patients (221.3 +/- 19.5 microg/l and 204.6 +/- 21.0 microg/l respectively). By using a specific protein-binding assay, we found that mean concentrations of IGF-II were 345.3 +/- 61.0 microg/l and 167.6 +/- 22.7 microg/l in tuberculous and malignant pleural effusions respectively. These values were significantly lower than those in serum of the same patients (628.3 +/- 79.0 microg/l, P

Stimulation of macrophages with endotoxin and/or cytokines is responsible for the expression of the inducible isoform of nitric oxide synthase (iNOS). Because macrophages are exposed to low pH within the microenvironment of inflammatory lesions, the potential role of low pH as an additional regulator of iNOS was investigated. Substitution of the culture medium of rat peritoneal macrophages at pH 7.4 with medium at pH 7.0 upregulated iNOS activity, as reflected by a 2.5-fold increase in nitrite accumulation. The increase in iNOS activity was associated with a similar increase in iNOS mRNA expression. Low environmental pH-induced iNOS gene expression involved the activation of nuclear factor-kappa B (NF-kappa B) transcription factor since [1] exposure of macrophages to low environmental pH increased NF-kappa B binding activity in the nucleus, and [2] treatment of macrophages with pyrrolidine dithiocarbamate or n-acetyl-leucinyl-norleucinal, two drugs preventing NF-kappa B translocation to the nucleus, canceled low pH-induced nitrite accumulation. The overall mechanism required the synthesis of tumor necrosis factor-alpha (TNF-alpha). Indeed, [1] elevated TNF-alpha bioactivity was observed in the medium of macrophages exposed to pH 7.0, and [2] incubation of macrophages with a neutralizing anti-TNF-alpha antibody impaired both NF-kappa B activation and nitrite accumulation in response to acid challenge. In summary, exposure of macrophages to acidic microenvironment in inflammatory lesions leads to the upregulation of iNOS activity through the activation of NF-kappa B.

OBJECTIVE: Acetylation or deacetylation of histone proteins may modulate cytokine gene transcription such as TNF alpha (TNF). We evaluated the balance between histone deacetytlase (HDAC) and histone acetyltransferase (HAT) in patients with rheumatoid arthritis (RA) or ankylosing spondylitis (AS) compared to healthy controls (HC) and determined the influence of HDAC inhibitors (trichostatin A -TSA- or Sirtinol -Sirt-) on these enzymatic activities and on the PBMC production of TNF. METHODS: 52 patients with RA, 21 with AS and 38 HC were evaluated. HAT and HDAC activities were measured on nuclear extracts from PBMC using colorimetric assays. Enzymatic activities were determined prior to and after ex vivo treatment of PBMC by TSA or Sirt. TNF levels were evaluated in PBMC culture supernatants in the absence or presence of TSA or Sirt. RESULTS: HAT and HDAC activities were significantly reduced in AS, while these activities reached similar levels in RA and HC. Ex vivo treatment of PBMC by HDACi tended to decrease HDAC expression in HC, but Sirt significantly reduced HAT in RA. TNF production by PBMC was significantly down-regulated by Sirt in HC and AS patients. CONCLUSION: HAT and HDAC were disturbed in AS while no major changes were found in RA. HDACi may modulate HDAC and HAT PBMC expression, especially Sirt in RA. Sirtinol was able to down regulate TNF production by PBMC in HC and AS. An imbalance between HAT and HDAC activities might provide the rationale for the development of HDACi in the therapeutic approach to inflammatory rheumatic diseases.

Background Sirtuin 1 (Sirt1) is a nuclear enzyme from the class III histone deacetylases that modulates gene expression and is involved in bone and cartilage remodeling. The goal of our study was to evaluate Sirt1 activity in peripheral blood mononuclear cells in patients with osteoarthritis in comparison with control patients, and to determine the relationship between Sirt1 activity and production of TNFα, IL-6 and IL-8 by peripheral blood mononuclear cells after ex vivo treatment with resveratrol, a Sirt1 activator. Results A prospective study was performed to compare the activity of Sirt1 in patients with primary osteoarthritis of the knee (American College of Rheumatology criteria) with its activity in controls. Peripheral blood mononuclear cells were isolated from peripheral blood, and Sirt1 activity evaluated from cytoplasmic and nuclear compartments using a fluorometric assay. Culture supernatant levels of TNFα, IL-6, and IL-8 were quantified before and after resveratrol ex vivo treatment. Nineteen patients with symptomatic knee osteoarthritis (age 64 ±9 years) and 18 controls (age 54 ±13 years) were included. No differences were found in cytoplasmic or nuclear Sirt1 activity between patients and controls. After resveratrol treatment, no changes in TNFα or IL-8 levels were found, but a significant dose-dependent increase in IL-6 levels was demonstrated in patients with osteoarthritis, but not controls. Sirt1 activity did not correlate with clinical activity (Lequesne’s index) or inflammation (erythrocyte sedimentation rate, C-reactive protein). Conclusion Sirt1 activity (cytoplasmic and nuclear) from peripheral blood mononuclear cells did not differ between patients with osteoarthritis and controls. Ex vivo treatment of peripheral blood mononuclear cells with resveratrol was associated with a dose-dependent increase in IL-6 levels only in patients with osteoarthritis.

Background Sirtuin 1 (Sirt1) is a nuclear enzyme from the class 3 histone deacetylases (HDACs) modulating gene expression, involved in the regulation of diverse biological processes ( cell survival, apoptosis, gluconeogenesis, adipogenesis, lipolysis), local and systemic inflammation, as well as in bone and cartilage remodeling. Objectives The main objective of the study was to evaluate Sirt1 activity in PBMC by venous blood aspiration in patients with osteoporosis (OP), compared to those of control patients, and to analyze the relationship between Sirt1 activity and production of mediators of inflammation and cytokines (TNFalpha, IL-6, IL-8) by the cells after ex vivo treatment with a Sirtuin activator, resveratrol, the most potent natural compound able to activate Sirt1. Methods A prospective and comparative monocentric study was performed in order to compare the activity of Sirt1 in patients with OP and controls. OP was defined by low BMD and at least one vertebral fracture. Secondary causes of osteoporosis were ruled out. PBMC were isolated from venous blood, and Sirt1 activity was evaluated from cytoplasmic and nuclear compartments using a fluorometric assay (SIRT1 fluorometric kit, BML-AK-555, Enzo Life Sciences, Villeurbanne, France) at the 15 minutes point. Culture supernatant levels of TNF alpha, IL-6, IL-8 were quantified before and after resveratrol ( 1 µmol and 5 µmol) ex vivo treatment, with commercial kits (Quantikine Kits, RD significance : p less than 0.05. Results Eighteen patients with symptomatic OP (age 68 ± 9 years) and 18 controls (age 54 ± 13 years) were included. No differences were found in cytoplasmic or nuclear Sirt1 activity between patients and controls. Cytoplasmic and nuclear Sirt1 activity were significantly correlated in patients and controls. Sirt1 activity did not correlate with bone biologic remodeling biomarkers or biologic markers of inflammation (CRP). After resveratrol treatment, no changes in TNF alpha, IL-6 or IL-8 levels were found, compared to pre treatment values, or between patients and controls. Conclusions Sirt1 activity (cytoplasmic and nuclear) from PBMC was not different between OP patients and controls. These results suggest that there is no implication of Sirt regulation of pro inflammatory cytokines or chemokines in PBMC from patients with osteoporosis. Disclosure of Interest None Declared

Background Sirtuin 1 (Sirt1) is a nuclear enzyme from the class 3 histone deacetylases (HDACs) modulating gene expression, involved in the regulation of diverse biological processes (cell survival, apoptosis, gluconeogenesis, adipogenesis, lipolysis), local and systemic inflammation, as well as in bone and cartilage remodeling. Objectives The main objective of the study was to evaluate Sirt1 activity in PBMC by venous blood aspiration in osteoarthritis (OA) patients, compared to those of control patients, and to analyze the relationship between Sirt1 activity and production of mediators of inflammation and cytokines (TNFalpha, IL-6, IL-8) by the cells after ex vivo treatment with a Sirtuin activator, resveratrol. Methods A prospective and comparative monocentric study was performed in order to compare the activity of Sirt1 in patients with osteoarthritis and controls. OA was defined according to the ACR criteria, with radiological grading (K-L); symptoms were quantified with Lequesne’s index. PBMC were isolated from venous blood, and Sirt1 activity was evaluated from cytoplasmic and nuclear compartments using a fluorometric assay (SIRT1 fluorimetric kit, BML-AK-555, Enzo Life Sciences, Villeurbanne, France) at the 15 minutes point. Culture supernatant levels of TNF alpha, IL-6, IL-8 were quantified before and after resveratrol (1 µmol and 5 µmol) ex vivo treatment, with commercial kits (Quantikine Kits, RD significance : p less than 0.05. Results Nineteen patients with symptomatic knee OA (age 64 ± 9 years, mean Lequesne’s index : 8.4; grade II or III of the K-L classification) and 18 controls (age 54 ± 13 years) were included. No differences were found in cytoplasmic or nuclear Sirt1 activity between patients and controls. Cytoplasmic and nuclear Sirt1 activity were correlated in patients and controls. Sirt1 activity (nuclear and cytoplasmic) was correlated to baseline IL-6 (p = 0.002) and baseline TNF (p = 0.004), but not with IL-8. Sirt1 activity did not correlate with clinical activity (Lequesne’s index) or biologic inflammation. After resveratrol treatment, no changes in TNF or IL-8 levels were found, but a significative, resveratrol-dose-dependent increase in IL-6 levels was demonstrated in OA patients (5.17 ± 0.89 pg/ml at baseline, 6,72 ± 1.31 after 1 µmol ex vivo treatment, and 7.64 ± 1.35 after 5 µmol ex vivo treatment; p = 0.02), not found in controls. Conclusions Sirt1 activity (cytoplasmic and nuclear) from PBMC was not different between OA patients and controls, nevertheless, ex vivo treatment of these PBMC with resveratrol, a Sirt1 activator, was unexpectedly associated with increased IL-6 levels in a dose-dependent relation only in OA patients, suggesting that IL-6 expression could be specifically regulated via Sirt1 in OA. Disclosure of Interest None Declared