The 10 Tesla muSR instrument: detector solutions

Solutions to the detector system of the High-Field muSR instrument at the Paul Scherrer Institut (PSI) in Switzerland are presented. The strict technical requirements are fulfilled through the application of Geiger-mode Avalanche Photodiodes.Comment: 6 pages, 4 figure

MEDIATION OF CHEMOTHERAPY-INDUCED APOPTOSIS BY THE LYSOSOMAL PROTEASE CATHEPSIN D

One of the most common hallmarks of cancer is dysregulation of cellular apoptotic processes. A comprehensive knowledge of the underlying mechanisms of the apoptotic machinery is vital for the identification of new drug targets and the development of innovative agents that stimulate the cell death process in cancer cells. Studies have shown that the lysosomal protease cathepsin D is important in the extrinsic apoptotic pathway stimulated by the death receptor ligands for TNFR1 and FAS, as well as by oxidative stress and the protein kinase C inhibitor staurosporine. To date, the role of cathepsin D in the chemotherapy-induced apoptotic pathway has not been characterized. This project examined the role of the lysosomal protease cathepsin D in chemotherapy-induced apoptosis of HeLa and U937 cells. The data demonstrated that following stimulation of U937 cells with the chemotherapy drug VP-16, cathepsin D was released into the cytosol approximately 4 hours after drug treatment. This release was selective for cathepsin D, as cathepsin B and the lysosomal markers LAMP and â-hexosaminidase were not released into the cytosol following VP-16 treatment. Inhibitors of caspases andcathepsin D had no effect on cathepsin D release, demonstrating that cathepsin D release occurred independently of caspase and cathepsin D activities. Downregulation of cathepsin D expression in U937 and Hela cells using siRNA was found to inhibit cell death resulting from a variety of stimuli, including death receptor ligands, oxidative stress, PKC inhibitors, and importantly, chemotherapy drugs. In addition, U937 and HeLa cells expressing cathepsin D siRNA exhibited delayed cytochrome c release and caspase-3 activation following VP-16 treatment. Moreover, isolated mitochondria from wild-type U937 cells released cytochrome c in response to cytosolic extracts that were treated with cathepsin D, suggesting that cathepsin D acts on a cytosolic factor to induce cytochrome c release. Inhibition of caspases had no impact on cytochrome c release provoked by cathepsin D-cleaved cytosolic extract, demonstrating that caspases are not mediators of cathepsin D-induced cytochrome c release. Taken together, these results demonstrate that cathepsin D is an important component of the apoptotic pathway and that it acts via an intermediary cytosolic factor to promote cytochrome c release and caspase activation during chemotherapy-induced apoptosis

The new versatile general purpose surface-muon instrument (GPS) based on silicon photomultipliers for μ{\mu}SR measurements on a continuous-wave beam

We report on the design and commissioning of a new spectrometer for muon-spin relaxation/rotation studies installed at the Swiss Muon Source (S$\mu$S) of the Paul Scherrer Institute (PSI, Switzerland). This new instrument is essentially a new design and replaces the old general-purpose surface-muon instrument (GPS) which has been for long the workhorse of the $\mu$SR user facility at PSI. By making use of muon and positron detectors made of plastic scintillators read out by silicon photomultipliers (SiPMs), a time resolution of the complete instrument of about 160 ps (standard deviation) could be achieved. In addition, the absence of light guides, which are needed in traditionally built $\mu$SR instrument to deliver the scintillation light to photomultiplier tubes located outside magnetic fields applied, allowed us to design a compact instrument with a detector set covering an increased solid angle compared to the old GPS.Comment: 11 pages, 11 figure

A time-resolution study with a plastic scintillator read out by a Geiger-mode Avalanche Photodiode

In this work we attempt to establish the best time resolution attainable with a scintillation counter consisting of a plastic scintillator read out by a Geiger-mode Avalanche Photodiode. The measured time resolution is inversely proportional to the square root of the energy deposited in the scintillator, and scales to 18ps (sigma) at 1MeV. This result competes with the best ones reported for photomultiplier tubes.Comment: 8 pages, 8 figure

Experiment K-6-09. Morphological and biochemical investigation of microgravity-induced nerve and muscle breakdown. Part 1: Investigation of nerve and muscle breakdown during spaceflight; Part 2: Biochemical analysis of EDL and PLT muscles

The present findings on rat hindlimb muscles suggest that skeletal muscle weakness induced by prolonged spaceflight can result from a combination of muscle fiber atrophy, muscle fiber segmental necrosis, degeneration of motor nerve terminals and destruction of microcirculatory vessels. Damage was confined to the red adductor longus (AL) and soleus muscles. The midbelly region of the AL muscle had more segmental necrosis and edema than the ends. Macrophages and neutrophils were the major mononucleated cells infiltrating and phagocytosing the cellular debris. Toluidine blue-positive mast cells were significantly decreased in Flight AL muscles compared to controls; this indicated that degranulation of mast cells contributed to tissue edema. Increased ubiquitination of disrupted myofibrils may have promoted myofilament degradation. Overall, mitochondria content and SDH activity were normal, except for a decrease in the subsarcolemmal region. The myofibrillar ATPase activity shifted toward the fast type in the Flight AL muscles. Some of the pathological changes may have occurred or been exacerbated during the 2 day postflight period of readaptation to terrestrial gravity. While simple atrophy should be reversible by exercise, restoration of pathological changes depends upon complex processes of regeneration by stem cells. Initial signs of muscle and nerve fiber regeneration were detected. Even though regeneration proceeds on Earth, the space environment may inhibit repair and cause progressive irreversible deterioration during long term missions. Muscles obtained from Flight rats sacrificed immediately (within a few hours) after landing are needed to distinguish inflight changes from postflight readaptation

Surface Tension Driven Convection Experiment Completed

The Surface Tension Driven Convection Experiment (STDCE) was designed to study basic fluid mechanics and heat transfer on thermocapillary flows generated by temperature variations along the free surfaces of liquids in microgravity. STDCE first flew on the USML-1 mission in July 1992 and was rebuilt for the USML-2 mission that was launched in October 1995. This was a collaborative project with principal investigators from Case Western Reserve University (CWRU), Professors Simon Ostrach and Yasuhiro Kamotani, along with a team from the NASA Lewis Research Center composed of civil servants and contractors from Aerospace Design & Fabrication, Inc. (ADF), Analex, and NYMA, Inc

Use of Fuzzycones for Sun-Only Attitude Determination: THEMIS Becomes ARTEMIS

In order for two THEMIS probes to successfully transition to ARTEMIS it will be necessary to determine attitudes with moderate accuracy using Sun sensor data only. To accomplish this requirement, an implementation of the Fuzzycones maximum likelihood algorithm was developed. The effect of different measurement uncertainty models on Fuzzycones attitude accuracy was investigated and a bin-transition technique was introduced to improve attitude accuracy using data with uniform error distributions. The algorithm was tested with THEMIS data and in simulations. The analysis results show that the attitude requirements can be met using Fuzzycones and data containing two bin-transitions

trans-complementation analysis of the flavivirus Kunjin ns5 gene reveals an essential role for translation of its N-terminal half in RNA replication

Recently we described rescue of defective Kunjin virus (KUN) RNAs with small deletions in the methyltransferase and RNA polymerase motifs of the ns5 gene, using BHK cells stably expressing KUN replicon RNA (repBHK cells) as helper (A. A. Khromykh et al., J. Virol. 72:7270-7279, 1998). We have now extended our previous observations and report successful trans-complementation of defective KUN RNAs with most of the ns5 gene deleted or substituted with a heterologous (dengue virus) ns5 sequence. Replication of full-length KUN RNAs with 3'-terminal deletions of 136 (5%), 933 (34%), and 1526 (56%) nucleotides in the ns5 gene was complemented efficiently in transfected repBHK cells. RNA with a larger deletion of 2,042 nucleotides (75%) was complemented less efficiently, and RNA with an even larger deletion of 2,279 nucleotides (84%) was not complemented at all. Chimeric KUN genomic RNA containing 87% of the KUN ns5 gene replaced by the corresponding sequence of the dengue virus type 2 ns5 gene was unable to replicate in normal BHK cells but was complemented in repBHK cells. These results demonstrate for the first time complementation of flavivirus RNAs with large deletions (as much as 75%) in the RNA polymerase gene and establish that translation of most of the N-terminal half of NS5 is essential for complementation in trans. A model of formation of the flavivirus replication complex implicating a possible role in RNA replication of conserved coding sequences in the N-terminal half of NS5 is proposed based on the complementation and earlier results with KUN and on reported data with other flaviviruses