Granulocyte-activating mediators (GRAM)

In the present study we investigated the capability of human epidermal cells to generate granulocyte-activating mediators (GRAM). It could be shown that human epidermal cells as well as an epidermoid carcinoma cell line (A431) produce an epidermal cell-derived granulocyte-activating mediator (EC-GRAM) which stimulates human granulocytes to release significant levels of toxic oxygen radicals as measured by a lucigenin-dependent chemiluminescence (CL). For further characterization of EC-GRAM the A431 cell line was used. Supernatants of A431 cells usually contained maximal EC-GRAM levels within 24 h of incubation. Factor production was enhanced by bacterial lipopolysaccharide (LPS), but not by silica particles and PHA. Moreover, freeze-thaw lysates of A431 cells and extracts of heat-separated human epidermis contained significant levels of EC-GRAM. Preincubation of granulocytes with EC-GRAM resulted in an enhanced response to subsequent stimulation with the chemotactic peptide f-met-phe. In contrast EC-GRAM did not affect the response to PMA or zymosan particles. However, EC-GRAM treated granulocytes were unresponsive to restimulation with EC-GRAM. Upon high performance liquid chromatography (HPLC) gel filtration EC-GRAM eluted within two major peaks exhibiting a molecular weight of 17 kD and 44 kD. According to its biochemical and biological properties EC-GRAM can be separated from other cytokines such as ETAF/-interleukin 1, interleukin 2, interferons, granulocyte colony-stimulating factor (G-CSF) and tumor necrosis factor (TNF). However, an antibody to human GM-CSF neutralized about 75% of the activity. These results indicate that EC-GRAM activity stimulating the generation of reactive oxygen species by granulocytes is probably due to GM-CSF

A study of flight control requirements for advanced, winged, earth-to-orbit vehicles with far-aft center-of-gravity locations

Control requirements of Controlled Configured Design Approach vehicles with far-aft center of gravity locations are studied. The baseline system investigated is a fully reusable vertical takeoff/horizontal landing single stage-to-orbit vehicle with mission requirements similar to that of the space shuttle vehicle. Evaluations were made to determine dynamic stability boundaries, time responses, trim control, operational center-of-gravity limits, and flight control subsystem design requirements. Study tasks included a baseline vehicle analysis, an aft center of gravity study, a payload size study, and a technology assessment

A turbojet-boosted two-stage-to-orbit space transportation system design study

The concept to use twin turbo-powered boosters for acceleration to supersonic staging speed followed by an all rocket powered orbiter stage was proposed. A follow-on design study was then made of the concept with the performance objective of placing a 29,483 Kg payload into a .2.6 X 195.3 km orbit. The study was performed in terms of analysis and trade studies, conceptual design, utility and economic analysis, and technology assessment. Design features of the final configuration included: strakes and area rule for improved take off and low transonic drag, variable area inlets, exits and turbine, and low profile fixed landing gear for turbojet booster stage. The payload required an estimated GLOW of 1,270,000 kg for injection in orbit. Each twin booster required afterburning turbojet engines each with a static sea level thrust rating of 444,800 N. Life cycle costs for this concept were comparable to a SSTO/SLED concept except for increased development cost due to the turbojet engine propulsion system

Applicability of the control configured design approach to advanced earth orbital transportation systems

The applicability of the control configured design approach (CCV) to advanced earth orbital transportation systems was studied. The baseline system investigated was fully reusable vertical take-off/horizontal landing single-stage-to-orbit vehicle and had mission requirements similar to the space shuttle orbiter. Technical analyses were made to determine aerodynamic, flight control and subsystem design characteristics. Figures of merit were assessed on vehicle dry weight and orbital payload. The results indicated that the major parameters for CCV designs are hypersonic trim, aft center of gravity, and control surface heating. Optimized CCV designs can be controllable and provide substantial payload gains over conventional non-CCV design vertical take-off vehicles

Human Granulocyte-Macrophage Colony Stimulating Factor: An Effective Direct Activator of Human Polymorphonuclear Neutrophilic Granulocytes

Granulocyet-macrophage colony-stimulating factor (GM-CSF) was shown to modulate different granulocyte functions. In the present study we investigated the effect of purified and recombinant human GM-CSF, particularly on the oxidative metabolism of isolated human granulocytes. In addition, ultrastructural changes upon stimulation were evaluated. For detection of granulocyte activation the following assay systems were used: 1) lucigenin-dependent chemiluminescence (CL),2) superoxide-dismutase (SOD) inhibitable cytochrome C-reduction (superoxide),3) horseradish peroxidase-mediated oxidation of phenol red (hydrogen peroxide),4) release of myeloperoxidase, 5) ultrastructural detection of hydrogen peroxide-production, and 6) scanning and transmission electron microscopy (SEM and TEM, respectively). A significant CL response was seen upon stimulation with recombinant human GM-CSF at concentrations ranging from 1 to 103 U/ml. The CL response started within 5-10 min with a maximum at 60 – 90 min and lasted more than 3 h. Thereafter granulocytes were completely deactivated to restimulation with the same mediator and with Tumor Necrosis Factor, but respondend to other triggers of the oxidative burst, whereas the response to f-met-leu-phe was significantly increased, The CL signal was completely blocked by an antiserum to GM-CSF. Moreover, the response was significantly inhibited by SOD and D-mannitol, suggesting the involvement of distinct reactive oxygen species (ROS) in generating the CL response. Significant amounts of superoxide were detected within 180 min after stimulation with GM-CSF, whereas, release of hydrogen peroxide and peroxidase were only minimal as shown by functional and ultrastructural assays. Activation of granulocytes could be visualized by SEM and TEM. GM-CSF stimulated cells showed an increased adherence to the substratum developing polarized filopodia and an increased number of intercellular vesicles within 30 min after addition of the stimulus. The results clearly demonstrate that GM-CSF directly stimulates granulocytes and, particularly, their oxidative metabolism. Therefore, GM-CSF which is probably released by epidermal cells appears to be a candidate for neutrophil activation in the skin, and thereby may play a crucial role in inflammatory skin diseases

Search for the Neutron Decay n→\rightarrow X+γ\gamma where X is a dark matter particle

In a recent paper submitted to Physical Review Letters, Fornal and Grinstein have suggested that the discrepancy between two different methods of neutron lifetime measurements, the beam and bottle methods can be explained by a previously unobserved dark matter decay mode, n$\rightarrow$ X+$\gamma$ where X is a dark matter particle. We have performed a search for this decay mode over the allowed range of energies of the monoenergetic gamma ray for X to be a dark matter particle. We exclude the possibility of a sufficiently strong branch to explain the lifetime discrepancy with greater than 4 sigma confidence.Comment: 6 pages 3 figure

Position-sensitive detection of ultracold neutrons with an imaging camera and its implications to spectroscopy

Position-sensitive detection of ultracold neutrons (UCNs) is demonstrated using an imaging charge-coupled device (CCD) camera. A spatial resolution less than 15 $\mu$m has been achieved, which is equivalent to an UCN energy resolution below 2 pico-electron-volts through the relation $\delta E = m_0g \delta x$. Here, the symbols $\delta E$, $\delta x$, $m_0$ and $g$ are the energy resolution, the spatial resolution, the neutron rest mass and the gravitational acceleration, respectively. A multilayer surface convertor described previously is used to capture UCNs and then emits visible light for CCD imaging. Particle identification and noise rejection are discussed through the use of light intensity profile analysis. This method allows different types of UCN spectroscopy and other applications.Comment: 12 figures, 28 pages, accepted for publication in NIM