High-speed Pellet Research Articles

Ignitor physics assessment and confinement projections

An independent assessment of the physics of Ignitor ispresented. Ignitor is a physics demonstration experiment with the main goalof achieving thermonuclear ignition, defined as the regime wherefusion alpha heating compensates for all forms of energy losses. Simulations show that a pulse of alpha particle power of up to 10-20 MW is produced for a period of more than a few seconds. Crucial issues are the production of peaked density profiles on atimescale of several energy confinement times, the control ofcurrent penetration for the optimization of ohmic heating andsawtooth avoidance. The presence of a 10-20 MW ICRF system and the operation of a high speed pellet injector are considered essential to provide added flexibility in order tocounter unexpected, adverse plasma behaviour.

Velocity Measurement and Freezing Technology of Multi-Shot Pellet on HL-1M Tokamak

A measuring system for high-speed hydrogen ice pellet was introduced in this paper and the photographing of flying pellet taken therein. With the system, a pellet (minimum size of Φ 0.3 mm) velocity (ranging from 50 m/s to 2500 m/s) can be measured in the HL-1M tokamak fueling experiments. By analyzing photographs and the conditions of frozen pellets (including gas supply, gas replenishment, temperature controlling etc), the pellet-freezing technology is summarized in the paper.

Heat Treatment Adaptations in Clostridium perfringens Vegetative Cells

Vegetative cells of Clostridium perfringens enterotoxigenic strains NCTC 8679, NCTC 8238, and H6 were grown at 37°C followed by a 60-min exposure to 28°C or 46°C. D10-values, as a measure of thermal resistance at 60°C, were significantly lower for 28°C exposures as compared with cultures given 37 and 46°C exposures. Following refrigeration at 4°C for 24 h, D10-values for the 37 and 46°C samples could not be differentiated from 28°C samples. Western immunoblot analyses of lysates from heat-adapted cells also detected the increased expression of proteins reacting with antiserum directed against the molecular chaperonins from Escherichia coli; GroEL, DnaJ, and the small acid soluble protein from Bacillus subtilis, SspC. Differential scanning calorimetry (DSC) identified thermal transitions corresponding to ribosomal protein denaturations at 72.1 ± 0.5°C. Any cellular heat adaptations in the DSC profiles were lost following refrigeration for several days to simulate minimally processed food storage conditions. Further analyses of high-speed pellets from crude cell extract fractions using two-dimensional gel electrophoresis detected the differential gene expression of at least four major proteins in heat-adapted vegetative cells of C. perfringens. N-terminal amino acid analyses identified two of the proteins as glyceraldehyde3-phosphate dehydrogenase and rubrerythrin. Both appear to have roles in this anaerobe under stressful conditions.

Refuelling performance improvement by high speed pellet launch from the magnetic high field side

Limited available pellet velocities have so far restricted the refuelling performance of efficient launch schemes from the tokamak magnetic high field side (HFS). Although pellet injection during H mode has resulted in more peaked density profiles and enhanced performance with respect to gas puff refuelling, prompt particle and energy losses caused by pellet induced ELM bursts have still limited the extension of the operational area. Now, the preliminary version of a new optimized pellet injection set-up at ASDEX Upgrade allows for significantly higher injection speeds when launching pellets from the magnetic HFS. Intact pellets with velocities up to vP = 560 m/s were successfully injected instead of the vP = 240 m/s available with the previous set-up. The velocity increase results in a deeper pellet penetration and seems to follow the vP1/3 scaling derived from a multimachine study using conventional pellet launch from the torus outside. The inward shift of the pellet particle deposition profile with respect to the penetration depths turned out to be approximately the same for both launch velocities. The respectively achieved deeper particle deposition inside the plasma column reduced the particle and energy loss rates during the immediate post-pellet phase. Thus, further enhancement of tokamak operation in the high density regime seems feasible by means of high speed pellets launched from the torus inner side.

Oxidative stress impairs insulin but not platelet-derived growth factor signalling in 3T3-L1 adipocytes.

Activation of phosphatidylinositol 3-kinase (PI 3-kinase) is a common event in both insulin and platelet-derived growth factor (PDGF) signalling, but only insulin activates this enzyme in the high-speed pellet (HSP), and induces GLUT4 translocation. Recently, we have demonstrated that exposure of 3T3-L1 adipocytes to oxidative stress impairs insulin-stimulated GLUT4 translocation and glucose transport, associated with impaired PI 3-kinase translocation and activation in the HSP [Tirosh, Potashnik, Bashan and Rudich (1999) J. Biol. Chem. 274, 10595-10602]. In this study the effect of a 2 h exposure to approximately 30 microM H(2)O(2) on insulin versus PDGF-BB signalling and metabolic effects was compared. PDGF-stimulated p85-associated PI 3-kinase activity in total cell lysates, as well as co-precipitation of the PDGF receptor, were unaffected by oxidative stress. Additionally, the increase in p85 association with the plasma-membrane lawns by PDGF remained intact following oxidation, whereas the insulin effect was decreased. PDGF significantly increased protein kinase B (PKB) activity in early differentiated cells, and that of p70 S6-kinase in both early and fully differentiated 3T3-L1 adipocytes. Following oxidation the effect of PDGF on PKB and p70 S6-kinase activation remained intact, whereas significant inhibition of insulin-stimulated activation of those enzymes was observed. In accordance, in both early and fully differentiated cells, oxidative stress completely blunted insulin- but not PDGF-stimulated protein synthesis. In conclusion, oxidative stress impairs insulin, but not PDGF, signalling and metabolic actions in both early and fully differentiated 3T3-L1 adipocytes. This emphasizes compartment-specific activation of PI 3-kinase as an oxidation-sensitive step specifically leading to insulin resistance.

Differential Protein Phosphorylation in 3T3-L1 Adipocytes in Response to Insulin VersusPlatelet-derived Growth Factor: NO EVIDENCE FOR A PHOSPHATIDYLINOSITIDE 3-KINASE-INDEPENDENT PATHWAY IN INSULIN SIGNALING

Insulin regulates glucose metabolism in adipocytes via a phosphatidylinositide 3-kinase (PI3K)-dependent pathway that appears to involve protein phosphorylation. However, the generation of phosphoinositides is not sufficient for insulin action, and it has been suggested that insulin regulation of glucose metabolism may involve both PI3K-dependent and -independent pathways, the latter being insulin specific. To test this hypothesis, we have designed a phosphoprotein screen to study insulin-specific phosphoproteins that may be either downstream or in parallel to PI3K. Nineteen insulin-regulated phosphospots were detected in the cytosol and high speed pellet fractions, only six of which were significantly regulated by platelet-derived growth factor. Importantly, almost all (92%) of the insulin-specific phosphoproteins identified using this approach were sensitive to the PI3K inhibitor wortmannin. Thus, we obtained no evidence for an insulin-specific, PI3K-independent signaling pathway. A large proportion (62%) of the insulin-specific phosphoproteins were enriched in the same high speed pellet fraction to which PI3K was recruited in response to insulin. Thus, our data suggest that insulin specifically stimulates the phosphorylation of a novel subset of downstream targets and this may in part be because of the unique localization of PI3K in response to insulin in adipocytes.

Mouse hepatitis virus replicase proteins associate with two distinct populations of intracellular membranes.

The coronavirus replicase gene (gene 1) is translated into two co-amino-terminal polyproteins that are proteolytically processed to yield more than 15 mature proteins. Several gene 1 proteins have been shown to localize at sites of viral RNA synthesis in the infected cell cytoplasm, notably on late endosomes at early times of infection. However, both immunofluorescence and electron microscopic studies have also detected gene 1 proteins at sites distinct from the putative sites of viral RNA synthesis or virus assembly. In this study, mouse hepatitis virus (MHV)-infected cells were fractionated and analyzed to determine if gene 1 proteins segregated to more than one membrane population. Following differential centrifugation of lysates of MHV-infected DBT cells, gene 1 proteins as well as the structural N and M proteins were detected almost exclusively in a high-speed small membrane pellet. Following fractionation of the small membrane pellet on an iodixanol density gradient, the gene 1 proteins p28 and helicase cofractionated with dense membranes (1.12 to 1.13 g/ml) that also contained peak concentrations of N. In contrast, p65 and p1a-22 were detected in a distinct population of less dense membranes (1.05 to 1.09 g/ml). Viral RNA was detected in membrane fractions containing helicase, p28, and N but not in the fractions containing p65 and p1a-22. LAMP-1, a marker for late endosomes and lysosomes, was detected in both membrane populations. These results demonstrate that multiple gene 1 proteins segregate into two biochemically distinct but tightly associated membrane populations and that only one of these populations appears to be a site for viral RNA synthesis. The results further suggest that p28 is a component of the viral replication complex whereas the gene 1 proteins p1a-22 and p65 may serve roles during infection that are distinct from viral RNA transcription or replication.

Release of Insulin Receptor Substrate Proteins from an Intracellular Complex Coincides with the Development of Insulin Resistance

Insulin receptor substrate (IRS) proteins are major substrates of the insulin receptor (IR). IRS-1 associates with an insoluble multiprotein complex, possibly the cytoskeleton, in adipocytes. This localization may facilitate interaction with the IR at the cell surface. In the present study, we examined the hypothesis that the release of IRS proteins from this location may be a mechanism for insulin desensitization. We show that a second IRS protein, IRS-2, is associated with a multiprotein complex in adipocytes with similar characteristics to the IRS-1 complex. Insulin treatment (15-60 min) caused the release of IRS-1 and IRS-2 from this complex (high speed pellet; HSP) into the cytosol, whereas the level of tyrosyl-phosphorylated IRS proteins remained constant. Chronic insulin treatment resulted in a dramatic reduction in IRS-1 and IRS-2 in the HSP, eventually (>2 h) leading to IRS protein degradation and decreased levels of tyrosyl-phosphorylated IRS proteins. Okadaic acid, which rapidly induces insulin resistance in adipocytes independently of IR function, caused an almost quantitative release of IRS-1 into the cytosol commensurate with a significant reduction in tyrosyl-phosphorylated IRS proteins. Platelet-derived growth factor, a factor known to compromise insulin signaling, caused a more moderate release of IRS proteins from the HSP. Collectively, these results suggest that the assembly of IRS-1/IRS-2 into a multiprotein complex facilitates coupling to the IR and that the regulated release from this location may represent a novel mechanism of insulin resistance.

Transport process of high-speed impurity pellets through curved guide tubes

Curved guide tubes to transport high-speed pellets are used for pellet injection into plasma devices. This article describes the transport process of the high-speed pellets through curved guide tubes. The pellets pass through the curved tubes with repeating reflections. Using the equation of friction at each reflection point, the velocity and the direction of the output pellet are calculated by following the pellet trajectory. The analytic expressions, with respect to the velocity and the direction of the output pellet, are also derived. It has been confirmed that there is good correspondence between the results obtained by following the pellet trajectories, those by the analytic expressions and those by the experiments. The analytic expressions are particularly useful for the design and making of the curved guide tubes used in the pellet injection.

Acceleration of large size deuterium pellets to high speeds using a small two-stage pneumatic gun

The technology of two-stage pneumatic pellet injectors represents by far the most reliable way to perform deep plasma fueling, with pipe gun devices capable of routinely launching small or medium size (up to 4 mm) D2 pellets at speeds in excess of 3 km/s, using rather small two-stage guns. It is still an open question, however, if scaling of the pellet size to the International Thermonuclear Experimental Reactor relevant values (6–8 mm) will or will not require a somewhat proportional increase in the physical size of the two-stage gun. In order to investigate this question, an extensive study was carried out at ENEA Frascati, using numerical simulation codes. It clearly indicated that a “compact” two-stage gun may have the potential to accelerate large size pellets at speeds up to 5 km/s. A low cost experiment was also scheduled. A spare pipe-gun cryostat of the single-shot two-stage pneumatic injector, previously used for high-speed pellet fueling of the Frascati tokamak upgrade, was modified in order to accommodate larger bore (up to 6 mm) launching barrels. In this article, we will mainly discuss the results of numerical simulations. A very early experimental campaign, carried out in 1996, will also be briefly reported, during which intact 6 mm D2 pellets were launched at speeds up to 2.5 km/s.

O3-induced inflammation in prepregnant, pregnant, and lactating rats correlates with O3 dose estimated by18O

Previous studies have shown that rats late in pregnancy and throughout lactation are more susceptible to ozone (O3)-induced pulmonary inflammation than are prepregnant (virgin) or postlactating rats. The major aim of the present study was to determine whether these differences in response intensity could be accounted for by the O3 dose to the lower region of the lung. The relative O3 dose to the lower lung of groups of pregnant, lactating, and virgin female rats was estimated by measuring the incorporation of the 18O isotope into low-speed (cells) and high-speed (surfactant) pellets of bronchoalveolar lavage fluid immediately after acute exposure to 0.5-1.1 parts/million 18O3. The polymorphonuclear leukocyte (PMN) and protein inflammatory responses were established 20 h after acute exposure of identical physiological groups to 0.5-1.1 parts/million 16O3 (common isotope). A single regression of PMN inflammation data against surfactant 18O concentration for all physiological groups gave a linear relationship, indicating direct proportionality of PMN inflammation with this estimate of relative dose to the lower lung regardless of physiological status. This implies that the chemical species that react with surfactant molecules, i.e., O3 or its metabolites, are the same as or proportional to those chemical species responsible for initiating PMN inflammation. Additional experiments showed that lung tissue ascorbic acid concentration was significantly lower in pregnant and lactating rats than in virgin female rats. Although a causative relationship cannot be assumed, the deficit in tissue ascorbic acid concentration in pregnant and lactating rats compared with virgin female rats is consistent with their greater responsiveness and higher relative surfactant O3 dose.

A Real-Time Protection System for the RFX Pellet Injector

In the RFX reversed field pinch (RFP) frozen H and D pellets with mass ≤5·1020 atoms and speed ≤1.5 km/s are used for plasma fuelling. If a high speed pellet is launched with no plasma or is not completely ablated, the graphite first wall may be damaged by its impact. The paper presents the design of a protection system, based on real-time measurements of plasma current and density, developed to prevent incomplete pellet ablation.

Sensory and Motor Neuron-derived Factor Is a Transmembrane Heregulin That Is Expressed on the Plasma Membrane with the Active Domain Exposed to the Extracellular Environment

The beta-heregulin sensory and motor neuron-derived factor (SMDF) has been suggested to be an important regulator of Schwann cell development and proliferation. In the present study, human SMDF was expressed in cultured cell lines. The cells and the recombinant protein were used to examine the membrane association and biological activity of the growth factor. Transfection of cells with SMDF cDNA constructs bearing FLAG epitope tags at either the amino- or carboxyl-terminal ends of the polypeptide resulted in expression of anti-FLAG immunoreactive polypeptides of approximately 44 and 83 kDa. The 83-kDa polypeptide was the major form expressed on the cell surface, as demonstrated by sensitivity to proteolysis in intact cells and surface biotinylation. SMDF was tightly associated with membranes isolated from transfected cells but was solubilized by Triton X-100. Immunofluorescent staining and immunoprecipitation experiments using cells expressing amino- or carboxyl-terminal tagged SMDF revealed that only the carboxyl-terminal end of the protein is exposed on the cell surface. Membranes from SMDF-transfected cells stimulated tyrosine phosphorylation of the beta-heregulin receptor ErbB3 in Schwann cells. Conditioned medium from transfected cells contained a similar activity, suggesting that SMDF is subject to proteolytic release from the plasma membrane. In contrast with other beta-heregulin isoforms, SMDF failed to bind heparin. Stimulation of Schwann cell ErbB3 receptor phosphorylation by SMDF was not affected by inhibition of Schwann cell heparan sulfate proteoglycan synthesis. These results demonstrate that SMDF is a type II transmembrane protein. This orientation places the active epidermal growth factor homology domain, which is located near the carboxyl-terminal end of the polypeptide, on the cell surface, where it can function as a membrane-anchored growth factor.

Comparative Studies of Glycoprotein Ib in Heparin Coated and Nonheparin Coated Extracorporeal Circulation Circuits

Contact between blood and artificial materials has various effects on blood. Impairment of platelet function is an especially important and well known effect, but its precise mechanism is not clearly understood. The authors constructed a circulation model to investigate the effect of extracorporeal circulation on platelet membrane glycoproteins (GPs), especially GP Ib, and to compare the changes in GP Ib in heparin coated (group C) and nonheparin coated (group N) circuits. As determined by flow cytometry, GP Ib in both groups decreased on initiating circulation, but the decrease in group N was significantly larger than that in group C. There was no observed change in GP IIb/IIIa levels in either group. The extent of shear stress induced platelet aggregation significantly decreased during circulation in both groups. Decreases in the extent of shear stress induced platelet aggregation were significantly less with the use of heparin coated circuits. In addition, the amount of GP Ib in the high speed pellet decreased progressively during circulation in both groups. In contrast, the amount of GP Ib in the Triton insoluble (low speed) pellet increased dramatically during circulation. However, expression of GP Ib in the Triton soluble platelet fraction was low in both groups. From the results, it was concluded that the cause of the decrease in platelet function during extracorporeal circulation is attributable to the internalization of GP Ib from the platelet surface inside the platelet. It also can be said that a heparin coated circuit is one effective means of controlling this change.

High-speed multishot pellet injector prototype for the Frascati Tokamak Upgrade

The Frascati Tokamak Upgrade (FTU) may require multiple high-speed pellet injection in order to achieve quasi-steady-state conditions. A research and development program was thus being pursued at ENEA Frascati, aimed at developing a multishot two-stage pellet injector (MPI), featuring eight “pipe gun” barrels and eight small two-stage pneumatic guns. According to FTU requirements, the final goal is to simultaneously produce up to eight D2 pellets, and then deliver them during a plasma pulse (1 s) with any time schedule, at speeds in the 1–2.5 km/s range. A prototype was constructed and tested to demonstrate the feasibility of the concept, and optimize pellet formation and firing sequences. This laboratory facility was automatically operated by means of a programmable logic controller (PLC), and had a full eight-shot capability. However, it was equipped as a first approach with only four two-stage guns. In this article we will describe in detail the guidelines of the MPI prototype design, which were strongly influenced by some external constraints. We will also report on the results of the experimental campaign, during which the feasibility of such a two-stage MPI was demonstrated. Sequences of four intact D2 pellets in the 1.2–1.6 mm size range, fired at time intervals of a few tens up to a few hundreds of ms, were routinely delivered in a laboratory experiment at injection speeds above 2.5 km/s, with good reproducibility and satisfactory aiming dispersion. Some preliminary effort to address the problem of propellant gas handling, based on an innovative approach, gave encouraging results, and work is in progress to carry out an experiment to definitely test the feasibility of this concept.

Intracellular localization of phosphatidylinositide 3-kinase and insulin receptor substrate-1 in adipocytes: potential involvement of a membrane skeleton.

Phosphatidylinositide (PI) 3-kinase binds to tyrosyl-phosphorylated insulin receptor substrate-1 (IRS-1) in insulin-treated adipocytes, and this step plays a central role in the regulated movement of the glucose transporter, GLUT4, from intracellular vesicles to the cell surface. PDGF, which also activates PI 3-kinase in adipocytes, has no significant effect on GLUT4 trafficking in these cells. We propose that this specificity may be mediated by differential localization of PI 3-kinase in response to insulin versus PDGF activation. Using subcellular fractionation in 3T3-L1 adipocytes, we show that insulin- and PDGF-stimulated PI 3-kinase activities are located in an intracellular high speed pellet (HSP) and in the plasma membrane (PM), respectively. The HSP is also enriched in IRS-1, insulin-stimulated tyrosyl-phosphorylated IRS-1 and intracellular GLUT4-containing vesicles. Using sucrose density gradient sedimentation, we have been able to segregate the HSP into two separate subfractions: one enriched in IRS-1, tyrosyl-phosphorylated IRS-1, PI 3-kinase as well as cytoskeletal elements, and another enriched in membranes, including intracellular GLUT4 vesicles. Treatment of the HSP with nonionic detergent, liberates all membrane constituents, whereas IRS-1 and PI 3-kinase remain insoluble. Conversely, at high ionic strength, membranes remain intact, whereas IRS-1 and PI 3-kinase become freely soluble. We further show that this IRS-1-PI 3-kinase complex exists in CHO cells overexpressing IRS-1 and, in these cells, the cytosolic pool of IRS-1 and PI 3-kinase is released subsequent to permeabilization with Streptolysin-O, whereas the particulate fraction of these proteins is retained. These data suggest that IRS-1, PI 3-kinase, as well as other signaling intermediates, may form preassembled complexes that may be associated with the actin cytoskeleton. This complex must be in close apposition to the cell surface, enabling access to the insulin receptor and presumably other signaling molecules that somehow confer the absolute specificity of insulin signaling in these cells.

Identification and localization of myosin phosphatase in human platelets.

Type 1 (PP1) and type 2A (PP2A) phosphatase activity was measured in three subcellular fractions of human platelets. About 80% of the activity was in the high-speed supernatant. Western blots showed that the catalytic subunit of PP1 (PP1c), including alpha- and delta-isoforms, was present in each fraction, but the level of the catalytic subunit of PP2A was very low in the low-speed pellet (cytoskeletal fraction). Various antibodies detected a subunit similar to the 130 kDa subunit (M130) of myosin phosphatase (MP) of smooth muscle in the low- and the high-speed pellets of human platelets. PP1c and associated proteins were isolated by microcystin-Sepharose. Many proteins were separated from each fraction, including myosin, actin and PP1c. M130 was separated only from the low-speed and the high-speed pellets. Kinase activities were detected in the unbound fractions, and fractions from the low- and high-speed pellets phosphorylated M130 and myosin respectively. Treatment of platelets with calyculin A increased the phosphorylation level of many proteins, including myosin heavy- and light-chains, and caused association of cytoskeletal proteins with the low-speed pellet. No marked change in the distribution of PP1c and M130 was detected. These results suggest that the MP in human platelets is composed of PP1c plus a subunit similar to M130 of the smooth muscle phosphatase.

Molecular cloning and transient expression in COS7 cells of a novel human PDE4B cAMP-specific phosphodiesterase, HSPDE4B3.

5'-Rapid amplification of cDNA ends, done on poly(A)+ RNA from human U87 cells, was used to identify 420 bp of novel 5' sequence of a PDE4B cAMP-specific phosphodiesterase (PDE). This identified an open reading frame encoding a putative 721-residue 'long-form' PDE4B splice variant, which we term HSPDE4B3. HSPDE4B3 differs from the two known PDE4B forms by virtue of its unique 79-residue N-terminal region, compared with the unique N-terminal regions of 94 and 39 residues found in HSPDE4B1 and HSPDE4B2 respectively. In transfected COS7 cells the two long forms, HSPDE4B1 and HSPDE4B3, had molecular masses of approx. 104 and approx. 103 kDa respectively. Expressed in COS-7 cells, the three HSPDE4B isoforms were found in the high-speed supernatant (cytosol) fraction as well as both the high-speed pellet (P2) and low-speed pellet (P1) fractions. All isoforms showed similar Km values for cAMP hydrolysis (1.5-2.6 microM). The maximal activities of the soluble cytosolic activity of the two long forms were very similar, whereas that of the short form, HSPDE4B2, was approx. 4-fold higher. Particulate-associated HSPDE4B1 and HSPDE4B2 were less active (approx. 40%) than their cytosol forms, whereas particulate HSPDE4B3 was similar in activity to its cytosolic form. Particulate and cytosolic forms of HSPDE4B1 and HSPDE4B3 were similarly inhibited by rolipram {4-[3-(cyclopentoxyl)-4-methoxyphenyl]-2-pyrrolidone}, the selective inhibitor of PDE4 (IC50 0.05-0.1 microM), whereas particulate-associated HSPDE4B2 was profoundly (approx. 10-fold) more sensitive (IC50 0.02 microM) to rolipram inhibition than its cytosolic form (IC50 0.2 microM). The various particulate-associated HSPDE4B isoforms showed very different susceptibilities to solubilization with the detergent Triton X-100 and high NaCl concentration. A novel cDNA, called pRPDE74, was obtained by screening a rat olfactory lobe cDNA library. This contained an open reading frame encoding a 721-residue protein that showed approx. 96% amino acid identity with HSPDE4B3 and is proposed to reflect the rat homologue of this human enzyme and is thus called RNPDE4B3. Alternative splicing of mRNA generated from both the human and rat PDE4B genes produces long and short splice variants that have unique N-terminal splice regions. It is suggested that these alternatively spliced regions determine changes in the maximal catalytic activity of the isoforms, their susceptibility to inhibition by rolipram and mode of interaction with particulate fractions.

Targeting of Tiam1 to the Plasma Membrane Requires the Cooperative Function of the N-terminal Pleckstrin Homology Domain and an Adjacent Protein Interaction Domain

The Rho-like GTPases Cdc42, Rac, and Rho play key roles in the regulation of the actin cytoskeleton and are implicated in transcriptional activation and cell transformation. We have previously identified the invasion-inducing Tiam1 gene, which encodes an activator of Rac. In fibroblasts, Tiam1 induces Rac-mediated membrane ruffling, which requires the N-terminal pleckstrin homology (PHn) domain. Here we show that this PHn domain is part of a protein interaction domain, which mediates membrane localization of Tiam1. After subcellular fractionation, up to 50% of Tiam1 is recovered in the Triton X-100-insoluble high speed pellet that contains small protein complexes. The regions in Tiam1 that are responsible for these protein interactions comprise the PHn domain, an adjacent putative coiled coil region (CC), and an additional flanking region (Ex). Deletions in each of these regions abolish membrane localization of Tiam1 and membrane ruffling, suggesting that they function cooperatively. Indeed, only polypeptides encompassing the PHn-CC-Ex region, and not the PHn-CC or the Ex region, localize at the membrane. These results indicate that the N-terminal PH domain is part of a larger functional Tiam1 domain that mediates protein complex formation and membrane localization of Tiam1.

High-speed hydrogen pellet acceleration using an electromagnetic railgun system

Using a low electric energy railgun system, solid hydrogen pellet acceleration tests have been conducted to investigate the application of the electromagnetic railgun system for high-speed pellet injection into fusion plasmas. Pneumatically pre-accelerated hydrogen pellets measuring 3 mm in diameter and 4–9 mm in length were successfully accelerated by a railgun system that uses a laser-induced plasma armature formation. A 2 m long single railgun with ceramic insulators accelerated the hydrogen pellet to 2.6 km s −1 with a supplied energy of 1.7 kJ. The average acceleration rate and the energy conversion coefficient were improved to about 1.6×10 6m s −2 and 0.37%, which is 1.6 times and three times as large as that using a railgun with plastic insulators, respectively. Furthermore, using the 1 m long augment railgun with ceramic insulators, the energy conversion coefficient was improved to about 0.55% while the acceleration rate was increased to 2.4×10 6 m s −2. The highest hydrogen pellet velocity attained was about 2.3 km s −1 for the augment railgun under an energy supply of 1.1 kJ. Based on the findings, it is expected that the acceleration efficiency and the pellet velocity can be further improved by using a longer augment railgun with ceramic insulators and by applying an optimal power supply.