17 research outputs found

    A PC program for estimating measurement uncertainty for aeronautics test instrumentation

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    A personal computer program was developed which provides aeronautics and operations engineers at Lewis Research Center with a uniform method to quickly provide values for the uncertainty in test measurements and research results. The software package used for performing the calculations is Mathcad 4.0, a Windows version of a program which provides an interactive user interface for entering values directly into equations with immediate display of results. The error contribution from each component of the system is identified individually in terms of the parameter measured. The final result is given in common units, SI units, and percent of full scale range. The program also lists the specifications for all instrumentation and calibration equipment used for the analysis. It provides a presentation-quality printed output which can be used directly for reports and documents

    High Response Dew Point Measurement System for a Supersonic Wind Tunnel

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    A new high response on-line measurement system has been developed to continuously display and record the air stream dew point in the NASA Lewis 10 x 10 supersonic wind tunnel. Previous instruments suffered from such problems as very slow response, erratic readings, and high susceptibility to contamination. The system operates over the entire pressure level range of the 10 x 10 SWT, from less than 2 psia to 45 psia, without the need for a vacuum pump to provide sample flow. The system speeds up tunnel testing, provides large savings in tunnel power costs and provides the dew point input for the data-reduction subroutines which calculate test section conditions

    Rotating pressure measurement system using an on board calibration standard

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    A computer-controlled multichannel pressure measurement system was developed to acquire detailed flow field measurements on board the Large Low Speed Centrifugal Compressor Research Facility at the NASA Lewis Research Center. A pneumatic slip ring seal assembly is used to transfer calibration pressures to a reference standard transducer on board the compressor rotor in order to measure very low differential pressures with the high accuracy required. A unique data acquisition system was designed and built to convert the analog signal from the reference transducer to the variable frequency required by the multichannel pressure measurement system and also to provide an output for temperature control of the reference transducer. The system also monitors changes in test cell barometric pressure and rotating seal leakage and provides an on screen warning to the operator if limits are exceeded. The methods used for the selection and testing of the the reference transducer are discussed, and the data acquisition system hardware and software design are described. The calculated and experimental data for the system measurement accuracy are also presented

    Cdc45 Limits Replicon Usage from a Low Density of preRCs in Mammalian Cells

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    Little is known about mammalian preRC stoichiometry, the number of preRCs on chromosomes, and how this relates to replicon size and usage. We show here that, on average, each 100-kb of the mammalian genome contains a preRC composed of approximately one ORC hexamer, 4–5 MCM hexamers, and 2 Cdc6. Relative to these subunits, ∼0.35 total molecules of the pre-Initiation Complex factor Cdc45 are present. Thus, based on ORC availability, somatic cells contain ∼70,000 preRCs of this average total stoichiometry, although subunits may not be juxtaposed with each other. Except for ORC, the chromatin-bound complement of preRC subunits is even lower. Cdc45 is present at very low levels relative to the preRC subunits, but is highly stable, and the same limited number of stable Cdc45 molecules are present from the beginning of S-phase to its completion. Efforts to artificially increase Cdc45 levels through ectopic expression block cell growth. However, microinjection of excess purified Cdc45 into S-phase nuclei activates additional replication foci by three-fold, indicating that Cdc45 functions to activate dormant preRCs and is rate-limiting for somatic replicon usage. Paradoxically, although Cdc45 colocalizes in vivo with some MCM sites and is rate-limiting for DNA replication to occur, neither Cdc45 nor MCMs colocalize with active replication sites. Embryonic metazoan chromatin consists of small replicons that are used efficiently via an excess of preRC subunits. In contrast, somatic mammalian cells contain a low density of preRCs, each containing only a few MCMs that compete for limiting amounts of Cdc45. This provides a molecular explanation why, relative to embryonic replicon dynamics, somatic replicons are, on average, larger and origin efficiency tends to be lower. The stable, continuous, and rate-limiting nature of Cdc45 suggests that Cdc45 contributes to the staggering of replicon usage throughout S-phase, and that replicon activation requires reutilization of existing Cdc45 during S-phase

    The Cytotoxic Necrotizing Factor of Yersinia pseudotuberculosis (CNFY) Enhances Inflammation and Yop Delivery during Infection by Activation of Rho GTPases.

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    Some isolates of Yersinia pseudotuberculosis produce the cytotoxic necrotizing factor (CNFY), but the functional consequences of this toxin for host-pathogen interactions during the infection are unknown. In the present study we show that CNFY has a strong influence on virulence. We demonstrate that the CNFY toxin is thermo-regulated and highly expressed in all colonized lymphatic tissues and organs of orally infected mice. Most strikingly, we found that a cnfY knock-out variant of a naturally toxin-expressing Y. pseudotuberculosis isolate is strongly impaired in its ability to disseminate into the mesenteric lymph nodes, liver and spleen, and has fully lost its lethality. The CNFY toxin contributes significantly to the induction of acute inflammatory responses and to the formation of necrotic areas in infected tissues. The analysis of the host immune response demonstrated that presence of CNFY leads to a strong reduction of professional phagocytes and natural killer cells in particular in the spleen, whereas loss of the toxin allows efficient tissue infiltration of these immune cells and rapid killing of the pathogen. Addition of purified CNFY triggers formation of actin-rich membrane ruffles and filopodia, which correlates with the activation of the Rho GTPases, RhoA, Rac1 and Cdc42. The analysis of type III effector delivery into epithelial and immune cells in vitro and during the course of the infection further demonstrated that CNFY enhances the Yop translocation process and supports a role for the toxin in the suppression of the antibacterial host response. In summary, we highlight the importance of CNFY for pathogenicity by showing that this toxin modulates inflammatory responses, protects the bacteria from attacks of innate immune effectors and enhances the severity of a Yersinia infection
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