High-Temperature, Thin-Film Strain Gages Improved

NASA Technical Reports Server (NTRS)

2005-01-01

Conventional resistance strain gage technology uses "bonded" strain gages. These foil or wire gages are bonded onto the surface of the test article with glue, ceramic cements, or flame-sprayed ceramics. These bonding agents can, in some instances, limit both the degree of strain transmission from the test structure to the gage and the maximum working temperature of the gage. Also, the bulky, bonded gage normally disrupts aerodynamic gas flow on the surface of the test structure because of its intrusive character. To respond to the urgent needs in aeronautic and aerospace research where stress and temperature gradients are high, aerodynamic effects need to be minimized, and higher operational temperatures are required, the NASA Lewis Research Center developed a thin film strain gage. This gage, a vacuum-deposited thin film formed directly on the surface of a test structure, operates at much higher temperatures than commercially available gages do and with minimal disruption of the aerodynamic flow. The gage uses an alloy, palladium-13 wt % chromium (hereafter, PdCr), which was developed by United Technologies Research Center under a NASA contract. PdCr is structurally stable and oxidation resistant up to at least 1100 C (2000 F); its temperature-induced resistance change is linear, repeatable, and not sensitive to the rates of heating and cooling. An early strain gage, which was made of 25-micrometer-diameter PdCr wire and demonstrated to be useable to 800 C, won an R&D 100 award in 1991. By further improving the purity of the material and by developing gage fabrication techniques that use sputter-deposition, photolithography patterning, and chemical etching, we have made an 8- to 10-m PdCr thin-film strain gage that can measure dynamic and static strain to at least 1100 C. For static strain measurements, a 5-m-thick Pt element serves as a temperature compensator to further minimize the temperature effect of the gage. These thin-film gages provide the advantage of

Inexpensive Bolt-Load Gage

NASA Technical Reports Server (NTRS)

Long, M. J.

1983-01-01

"Built-in" gage determines whether large bolt or stud has been torqued to desired load and provides for continuous inspection to ensure proper load is being maintained. Gage detects longitudinal stress/strain bolt; requires no electronic or sonic test equipment.

Palladium-chromium static strain gages for high temperatures

NASA Technical Reports Server (NTRS)

Lei, Jih-Fen

1992-01-01

An electrical resistance strain gage that can provide accurate static strain measurement to a temperature of 1500 F or above is being developed both in fine wire and thin film forms. The gage is designed to be temperature compensated on any substrate material. It has a dual element: the gage element is a special alloy, palladium-13wt percent chromium (PdCr), and the compensator element is platinum (Pt). Earlier results of a PdCr based wire gage indicated that the apparent strain of this gage can be minimized and the repeatability of the apparent strain can be improved by prestabilizing the gage on the substrate for a long period of time. However, this kind of prestabilization is not practical in many applications and therefore the development of a wire gage which is prestabilized before installation on the substrate is desirable. This paper will present our recent progress in the development of a prestabilized wire gage which can provide meaningful strain data for the first thermal cycle. A weldable PdCr gage is also being developed for field testing where conventional flame-spraying installation can not be applied. This weldable gage is narrower than a previously reported gage, thereby allowing the gage to be more resistant to buckling under compressive loads. Some preliminary results of a prestabilized wire gage flame-sprayed directly on IN100, an engine material, and a weldable gage spot-welded on IN100 and SCS-6/(beta)21-S Titanium Matrix Composite (TMC), a National Aero-Space Plane (NASP) structure material, will be reported. Progress on the development of a weldable thin film gage will also be addressed. The measurement technique and procedures and the lead wire effect will be discussed.

Elevated temperature strain gages

NASA Technical Reports Server (NTRS)

Brittain, J. O.; Geslin, D.; Lei, J. F.

1985-01-01

Materials were evaluated that could be used in manufacturing electrical resistance strain gages for static strain measurements at temperatures at or above 1273 K. Strain gage materials must have a characteristic response to strain, temperature and time that is reproducible or that varies in a predictable manner within specified limits. Several metallic alloys were evaluated, as well as a series of transition metal carbides, nitrides and silicides.

Gage for 3-d contours

NASA Technical Reports Server (NTRS)

Haynie, C. C.

1980-01-01

Simple gage, used with template, can help inspectors determine whether three-dimensional curved surface has correct contour. Gage was developed as aid in explosive forming of Space Shuttle emergency-escape hatch. For even greater accuracy, wedge can be made of metal and calibrated by indexing machine.

GAGES: A stream gage database for evaluating natural and alteredflow conditions in the conterminous United States

USGS Publications Warehouse

Falcone, James A.; Carlisle, Daren M.; Wolock, David M.; Meador, Michael R.

2010-01-01

In addition, watersheds were assessed for their reference quality within nine broad regions for use in studies intended to characterize stream flows under conditions minimally influenced by human activities. Three primary criteria were used to assess reference quality: (1) a quantitative index of anthropogenic modification within the watershed based on GIS-derived variables, (2) visual inspection of every stream gage and drainage basin from recent high-resolution imagery and topographic maps, and (3) information about man-made influences from USGS Annual Water Data Reports. From the set of 6785 sites, we identified 1512 as reference-quality stream gages. All data derived for these watersheds as well as the reference condition evaluation are provided as an online data set termed GAGES (geospatial attributes of gages for evaluating stream flow).

Inflatable device for installing strain gage bridges

NASA Technical Reports Server (NTRS)

Cook, C. E.; Smith, G. E.; Monaghan, R. C. (Inventor)

1983-01-01

Methods and devices for installing in a tubular shaft multiple strain gages are disclosed with focus on a method and a device for pneumatically forcing strain gages into seated engagement with the internal surfaces of a tubular shaft in an installation of multiple strain gages in a tubular shaft. The strain gages or other electron devices are seated in a template-like component which is wrapped about a pneumatically expansible body. The component is inserted into a shaft and the body is pneumatically expanded after a suitable adhesive was applied to the surfaces.

Evaluation of the trough-type rain gage

Treesearch

Irvin C. Reigner

1964-01-01

Although infrequently used, the trough-type rain gage has been described as aerodynamically the most efficient of gages (Hayes and Kittredge 1949). This applies only to the commercial semicircular eave-gutter type of trough and not to troughs rectangular in cross-section. The latter, as well as the upright cylindrical type of gage, produces greater disturbances in the...

Sensitivity of hot-cathode ionization vacuum gages in several gases

NASA Technical Reports Server (NTRS)

Holanda, R.

1972-01-01

Four hot-cathode ionization vacuum gages were calibrated in 12 gases. The relative sensitivities of these gages were compared to several gas properties. Ionization cross section was the physical property which correlated best with gage sensitivity. The effects of gage accelerating voltage and ionization-cross-section energy level were analyzed. Recommendations for predicting gage sensitivity according to gage type were made.

Triple-material stress-strain resistivity gage

DOEpatents

Stout, R.B.

1987-05-19

A triple material piezoresistive gage provides multi-component elastic stress or strain measurements. Thin foils of three piezoresistive materials, e.g., ytterbium, manganin, and constantan, are configured in a nested serpentine rectilinear grind or other grind arrangement and embedded in a medium, preferably normal to the direction of shock wave propagation. The output of the gage is a resistivity change history for each material of gage. Each resistivity change is independent of the others so that three diagonal components of the elastic stress or strain tensor can be calculated form the resistivity measurements. 4 figs.

Triple-material stress-strain resistivity gage

DOEpatents

Stout, Ray B.

1988-01-01

A triple material piezoresistive gage provides multi-component elastic stress or measurements. Thin foils of three piezoresistive materials, e.g. ytterbium, manganin, and constantan, are configured in a nested serpentine rectilinear grid or other grid arrangement and embedded in a medium, preferably normal to the direction of shock wave propagation. The output of the gage is a resistivity change history for each material of the gage. Each resistivity change is independent of the others so that three diagonal components of the elastic stress or strain tensor can be calculated from the resistivity measurements.

Strain gage based determination of mixed mode SIFs

NASA Astrophysics Data System (ADS)

Murthy, K. S. R. K.; Sarangi, H.; Chakraborty, D.

2018-05-01

Accurate determination of mixed mode stress intensity factors (SIFs) is essential in understanding and analysis of mixed mode fracture of engineering components. Only a few strain gage determination of mixed mode SIFs are reported in literatures and those also do not provide any prescription for radial locations of strain gages to ensure accuracy of measurement. The present investigation experimentally demonstrates the efficacy of a proposed methodology for the accurate determination of mixed mode I/II SIFs using strain gages. The proposed approach is based on the modified Dally and Berger's mixed mode technique. Using the proposed methodology appropriate gage locations (optimal locations) for a given configuration have also been suggested ensuring accurate determination of mixed mode SIFs. Experiments have been conducted by locating the gages at optimal and non-optimal locations to study the efficacy of the proposed approach. The experimental results from the present investigation show that highly accurate SIFs (0.064%) can be determined using the proposed approach if the gages are located at the suggested optimal locations. On the other hand, results also show the very high errors (212.22%) in measured SIFs possible if the gages are located at non-optimal locations. The present work thus clearly substantiates the importance of knowing the optimal locations of the strain gages apriori in accurate determination of SIFs.

Elevated temperature strain gages

NASA Technical Reports Server (NTRS)

Brittain, J. O.; Geslin, D.; Lei, J. F.

1986-01-01

One of the goals of the HOST Program is the development of electrical resistance strain gages for static strain measurements at temperatures equal to or greater than 1273 K. Strain gage materials must have a reproducible or predictable response to temperature, time and strain. It is the objective of this research to investigate criteria for the selection of materials for such applications through electrical properties studies. The results of the investigation of two groups of materials, refractory compounds and binary alloy solid solutions are presented.

Bi-directional, buried-wire skin-friction gage

NASA Technical Reports Server (NTRS)

Higuchi, H.; Peake, D. J.

1978-01-01

A compact, nonobtrusive, bi-directional, skin-friction gage was developed to measure the mean shear stress beneath a three-dimensional boundary layer. The gage works by measuring the heat flux from two orthogonal wires embedded in the surface. Such a gage was constructed and its characteristics were determined for different angles of yaw in a calibration experiment in subsonic flow with a Preston tube used as a standard. Sample gages were then used in a fully three-dimensional turbulent boundary layer on a circular cone at high relative incidence, where there were regimes of favorable and adverse pressure gradients and three-dimensional separation. Both the direction and magnitude of skin friction were then obtained on the cone surface.

Triple-material stress-strain resistivity gage

DOEpatents

Stout, R.B.

1988-05-17

A triple material piezoresistive gage provides multi-component elastic stress measurements is disclosed. Thin foils of three piezoresistive materials, e.g. ytterbium, manganin, and constantan, are configured in a nested serpentine rectilinear grid or other grid arrangement and embedded in a medium, preferably normal to the direction of shock wave propagation. The output of the gage is a resistivity change history for each material of the gage. Each resistivity change is independent of the others so that three diagonal components of the elastic stress or strain tensor can be calculated from the resistivity measurements. 4 figs.

Telescoping magnetic ball bar test gage

DOEpatents

Bryan, J.B.

1984-03-13

A telescoping magnetic ball bar test gage for determining the accuracy of machine tools, including robots, and those measuring machines having non-disengageable servo drives which cannot be clutched out is disclosed. Two gage balls are held and separated from one another by a telescoping fixture which allows them relative radial motional freedom but not relative lateral motional freedom. The telescoping fixture comprises a parallel reed flexure unit and a rigid member. One gage ball is secured by a magnetic socket knuckle assembly which fixes its center with respect to the machine being tested. The other gage ball is secured by another magnetic socket knuckle assembly which is engaged or held by the machine in such manner that the center of that ball is directed to execute a prescribed trajectory, all points of which are equidistant from the center of the fixed gage ball. As the moving ball executes its trajectory, changes in the radial distance between the centers of the two balls caused by inaccuracies in the machine are determined or measured by a linear variable differential transformer (LVDT) assembly actuated by the parallel reed flexure unit. Measurements can be quickly and easily taken for multiple trajectories about several different fixed ball locations, thereby determining the accuracy of the machine. 3 figs.

Telescoping magnetic ball bar test gage

DOEpatents

Bryan, J.B.

1982-03-15

A telescoping magnetic ball bar test gage for determining the accuracy of machine tools, including robots, and those measuring machines having non-disengagable servo drives which cannot be clutched out. Two gage balls are held and separated from one another by a telescoping fixture which allows them relative radial motional freedom but not relative lateral motional freedom. The telescoping fixture comprises a parallel reed flexure unit and a rigid member. One gage ball is secured by a magnetic socket knuckle assembly which fixes its center with respect to the machine being tested. The other gage ball is secured by another magnetic socket knuckle assembly which is engaged or held by the machine in such manner that the center of that ball is directed to execute a prescribed trajectory, all points of which are equidistant from the center of the fixed gage ball. As the moving ball executes its trajectory, changes in the radial distance between the centers of the two balls caused by inaccuracies in the machine are determined or measured by a linear variable differential transformer (LVDT) assembly actuated by the parallel reed flexure unit. Measurements can be quickly and easily taken for multiple trajectories about several different fixed ball locations, thereby determining the accuracy of the machine.

49 CFR 179.400-19 - Valves and gages.

Code of Federal Regulations, 2014 CFR

2014-10-01

... control of vapor phase pressure, vapor phase venting, liquid transfer and liquid flow rates. All valves... within suitable protective housings. A liquid level gage and a vapor phase pressure gage must be provided... Liquid Tank Car Tanks and Seamless Steel Tanks (Classes DOT-113 and 107A) § 179.400-19 Valves and gages...

49 CFR 179.400-19 - Valves and gages.

Code of Federal Regulations, 2012 CFR

2012-10-01

... control of vapor phase pressure, vapor phase venting, liquid transfer and liquid flow rates. All valves... within suitable protective housings. A liquid level gage and a vapor phase pressure gage must be provided... Liquid Tank Car Tanks and Seamless Steel Tanks (Classes DOT-113 and 107A) § 179.400-19 Valves and gages...

49 CFR 179.400-19 - Valves and gages.

Code of Federal Regulations, 2013 CFR

2013-10-01

... control of vapor phase pressure, vapor phase venting, liquid transfer and liquid flow rates. All valves... within suitable protective housings. A liquid level gage and a vapor phase pressure gage must be provided... Liquid Tank Car Tanks and Seamless Steel Tanks (Classes DOT-113 and 107A) § 179.400-19 Valves and gages...

Stage measurement at gaging stations

USGS Publications Warehouse

Sauer, Vernon B.; Turnipseed, D. Phil

2010-01-01

Stream and reservoir stage are critical parameters in the computation of stream discharge and reservoir volume, respectively. In addition, a record of stream stage is useful in the design of structures that may be affected by stream elevation, as well as for the planning for various uses of flood plains. This report describes equipment and methodology for the observation, sensing, and recording of stage in streams and reservoirs. Although the U.S. Geological Survey (USGS) still uses the traditional, basic stilling-well float system as a predominant gaging station, modern electronic stage sensors and water-level recorders are now commonly used. Bubble gages coupled with nonsubmersible pressure transducers eliminate the need for stilling wells. Submersible pressure transducers have become common in use for the measurement of stage in both rivers and lakes. Furthermore, noncontact methods, such as radar, acoustic, and laser methods of sensing water levels, are being developed and tested, and in the case of radar, are commonly used for the measurement of stage. This report describes commonly used gaging-station structures, as well as the design and operation of gaging stations. Almost all of the equipment and instruments described in this report will meet the accuracy standard set by the USGS Office of Surface Water (OSW) for the measurement of stage for most applications, which is ±0.01 foot (ft) or 0.2 percent of the effective stage. Several telemetry systems are used to transmit stage data from the gaging station to the office, although satellite telemetry has become the standard. These telemetry systems provide near real-time stage data, as well as other information that alerts the hydrographer to extreme or abnormal events, and instrument malfunctions.

Experiments with windshields for precipitation gages

Treesearch

C. C. Warnick

1953-01-01

Under cooperative encouragement from several Federal agencies, the Engineering Experiment Station of the University of Idaho has been studying the principles, development, and use of high-altitude precipitation gages. A low-speed wind tunnel has been used to study the effect of wind on the catching characteristics of model storage precipitation gages. A snow storm was...

Telescoping magnetic ball bar test gage

DOEpatents

Bryan, James B.

1984-01-01

A telescoping magnetic ball bar test gage for determining the accuracy of machine tools, including robots, and those measuring machines having non-disengageable servo drives which cannot be clutched out. Two gage balls (10, 12) are held and separated from one another by a telescoping fixture which allows them relative radial motional freedom but not relative lateral motional freedom. The telescoping fixture comprises a parallel reed flexure unit (14) and a rigid member (16, 18, 20, 22, 24). One gage ball (10) is secured by a magnetic socket knuckle assembly (34) which fixes its center with respect to the machine being tested. The other gage ball (12) is secured by another magnetic socket knuckle assembly (38) which is engaged or held by the machine in such manner that the center of that ball (12) is directed to execute a prescribed trajectory, all points of which are equidistant from the center of the fixed gage ball (10). As the moving ball (12) executes its trajectory, changes in the radial distance between the centers of the two balls (10, 12) caused by inaccuracies in the machine are determined or measured by a linear variable differential transformer (LVDT) assembly (50, 52, 54, 56, 58, 60) actuated by the parallel reed flexure unit (14). Measurements can be quickly and easily taken for multiple trajectories about several different fixed ball (10) locations, thereby determining the accuracy of the machine.

Strain gage installation and survivability on geosynthetics used in flexible pavements

NASA Astrophysics Data System (ADS)

Brooks, Jeremy A.

The use of foil type strain gages on geosynthetics is poorly documented. In addition, very few individuals are versed in proper installation techniques or calibration methods. Due to the limited number of knowledgeable technicians there is no information regarding the susceptibility of theses gages to errors in installation by inexperienced installers. Also lacking in the documentation related to the use of foil type strain gages on geosynthetics is the survivability of the gages in field conditions. This research documented procedures for installation, calibration, and survivability used by the project team to instruments a full scale field installation in Marked Tree, AR. This research also addressed sensitivity to installation errors on both geotextile and geogrid. To document the process of gage installation an experienced technician, Mr. Joe Ables, formerly of the UASCE Waterways Experiment Station, was consulted. His techniques were combined with those discovered in related literature and those developed by the research team to develop processes that were adaptable to multiple gage geometries and parent geosynthetics. These processes were described and documented in a step by step manner with accompanying photographs, which should allow virtually anyone with basic electronics knowledge to install these gages properly. Calibration of the various geosynthetic / strain gage combinations was completed using wide width tensile testing on multiple samples of each material. The tensile testing process was documented and analyzed using digital photography to analyze strain on the strain gage itself. Calibration factors for each geosynthtics used in the full scale field testing were developed. In addition, the process was thoroughly documented to allow future researchers to calibrate additional strain gage and geosynthetic combinations. The sensitivity of the strain gages to installation errors was analyzed using wide width tensile testing and digital photography to

Microprocessor-based multichannel flutter monitor using dynamic strain gage signals

NASA Technical Reports Server (NTRS)

Smalley, R. R.

1976-01-01

Two microprocessor-based multichannel monitors for monitoring strain gage signals during aerodynamic instability (flutter) testing in production type turbojet engines were described. One system monitors strain gage signals in the time domain and gives an output indication whenever the signal amplitude of any gage exceeds a pre-set alarm or abort level for that particular gage. The second system monitors the strain gage signals in the frequency domain and therefore is able to use both the amplitude and frequency information. Thus, an alarm signal is given whenever the spectral content of the strain gage signal exceeds, at any point, its corresponding amplitude vs. frequency limit profiles. Each system design is described with details on design trade-offs, hardware, software, and operating experience.

Gage for measuring coastal erosion and sedimentation

NASA Technical Reports Server (NTRS)

Carpini, T. D.; Moughon, W. C.

1970-01-01

Underwater sand height gage, which measures heights up to 12 inches, is comprised of two standard flush-diaphragm pressure transducers. Gage is very sensitive to buried water heights and is useful as a research tool in study of wet earth and landslide phenomena.

Evaluation results of the 700 deg C Chinese strain gages

NASA Technical Reports Server (NTRS)

Hobart, H. F.

1984-01-01

There is a continuing interest and need for resistance strain gages capable of making static strain measurements on components located in the hot section of gas turbine engines. A paper by Tsen-tai Wu describes the development and evaluation of high temperature gauges fabricated from specially developed Fe-Cr-Al-V-Ti-Y alloy wire. Several of these gages and a quantity of P12-2 ceramic adhesive were purchased for evaluation. Nine members of the aircraft turbine engine community were invited to participate in an evaluation of these gages. Each participant was sent one strain gage, a small amount of ceramic adhesive, instructions for mounting the gage on a test beam, and a set of suggestions for the experiment. Data on gage factor variation with temperature, apparent strain, and drift are discussed.

Bonding of strain gages to fiber reinforced composite plastic materials

NASA Technical Reports Server (NTRS)

Chamis, C. C.; Hanson, M. P.; Serafini, T. T.

1970-01-01

Strain gage is installed during molding of composite and utilizes the adhesive properties of the matrix resin in the composite to bond the strain gage in place. Gages thus embedded provide data at all temperatures that the matrix can withstand.

Cost effectiveness of the stream-gaging program in Louisiana

USGS Publications Warehouse

Herbert, R.A.; Carlson, D.D.

1985-01-01

This report documents the results of a study of the cost effectiveness of the stream-gaging program in Louisiana. Data uses and funding sources were identified for the 68 continuous-record stream gages currently (1984) in operation with a budget of $408,700. Three stream gages have uses specific to a short-term study with no need for continued data collection beyond the study. The remaining 65 stations should be maintained in the program for the foreseeable future. In addition to the current operation of continuous-record stations, a number of wells, flood-profile gages, crest-stage gages, and stage stations, are serviced on the continuous-record station routes; thus, increasing the current budget to $423,000. The average standard error of estimate for data collected at the stations is 34.6%. Standard errors computed in this study are one measure of streamflow errors, and can be used as guidelines in comparing the effectiveness of alternative networks. By using the routes and number of measurements prescribed by the ' Traveling Hydrographer Program, ' the standard error could be reduced to 31.5% with the current budget of $423,000. If the gaging resources are redistributed, the 34.6% overall level of accuracy at the 68 continuous-record sites and the servicing of the additional wells or gages could be maintained with a budget of approximately $410,000. (USGS)

Cost effectiveness of the stream-gaging program in Ohio

USGS Publications Warehouse

Shindel, H.L.; Bartlett, W.P.

1986-01-01

This report documents the results of the cost effectiveness of the stream-gaging program in Ohio. Data uses and funding sources were identified for 107 continuous stream gages currently being operated by the U.S. Geological Survey in Ohio with a budget of $682,000; this budget includes field work for other projects and excludes stations jointly operated with the Miami Conservancy District. No stream gage were identified as having insufficient reason to continue their operation; nor were any station identified as having uses specifically only for short-term studies. All 107 station should be maintained in the program for the foreseeable future. The average standard error of estimation of stream flow records is 29.2 percent at its present level of funding. A minimum budget of $679,000 is required to operate the 107-gage program; a budget less than this does no permit proper service and maintenance of the gages and recorders. At the minimum budget, the average standard error is 31.1 percent The maximum budget analyzed was $1,282,000, which resulted in an average standard error of 11.1 percent. A need for additional gages has been identified by the other agencies that cooperate in the program. It is suggested that these gage be installed as funds can be made available.

NASA LaRC Strain Gage Balance Design Concepts

NASA Technical Reports Server (NTRS)

Rhew, Ray D.

1999-01-01

The NASA Langley Research Center (LaRC) has been designing strain-gage balances for more than fifty years. These balances have been utilized in Langley's wind tunnels, which span over a wide variety of aerodynamic test regimes, as well as other ground based test facilities and in space flight applications. As a result, the designs encompass a large array of sizes, loads, and environmental effects. Currently Langley has more than 300 balances available for its researchers. This paper will focus on the design concepts for internal sting mounted strain-gage balances. However, these techniques can be applied to all force measurement design applications. Strain-gage balance concepts that have been developed over the years including material selection, sting, model interfaces, measuring, sections, fabrication, strain-gaging and calibration will be discussed.

Resistance fail strain gage technology as applied to composite materials

NASA Technical Reports Server (NTRS)

Tuttle, M. E.; Brinson, H. F.

1985-01-01

Existing strain gage technologies as applied to orthotropic composite materials are reviewed. The bonding procedures, transverse sensitivity effects, errors due to gage misalignment, and temperature compensation methods are addressed. Numerical examples are included where appropriate. It is shown that the orthotropic behavior of composites can result in experimental error which would not be expected based on practical experience with isotropic materials. In certain cases, the transverse sensitivity of strain gages and/or slight gage misalignment can result in strain measurement errors.

49 CFR 213.355 - Frog guard rails and guard faces; gage.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 4 2014-10-01 2014-10-01 false Frog guard rails and guard faces; gage. 213.355... Higher § 213.355 Frog guard rails and guard faces; gage. The guard check and guard face gages in frogs... distance between the gage line of a frog to the guard line 1 of its guard rail or guarding face, measured...

49 CFR 213.355 - Frog guard rails and guard faces; gage.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 49 Transportation 4 2012-10-01 2012-10-01 false Frog guard rails and guard faces; gage. 213.355... Higher § 213.355 Frog guard rails and guard faces; gage. The guard check and guard face gages in frogs... distance between the gage line of a frog to the guard line 1 of its guard rail or guarding face, measured...

49 CFR 213.355 - Frog guard rails and guard faces; gage.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 49 Transportation 4 2013-10-01 2013-10-01 false Frog guard rails and guard faces; gage. 213.355... Higher § 213.355 Frog guard rails and guard faces; gage. The guard check and guard face gages in frogs... distance between the gage line of a frog to the guard line 1 of its guard rail or guarding face, measured...

49 CFR 213.355 - Frog guard rails and guard faces; gage.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 49 Transportation 4 2011-10-01 2011-10-01 false Frog guard rails and guard faces; gage. 213.355... Higher § 213.355 Frog guard rails and guard faces; gage. The guard check and guard face gages in frogs... distance between the gage line of a frog to the guard line 1 of its guard rail or guarding face, measured...

49 CFR 213.355 - Frog guard rails and guard faces; gage.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 4 2010-10-01 2010-10-01 false Frog guard rails and guard faces; gage. 213.355... Higher § 213.355 Frog guard rails and guard faces; gage. The guard check and guard face gages in frogs... distance between the gage line of a frog to the guard line 1 of its guard rail or guarding face, measured...

Cost effectiveness of the stream-gaging program in Pennsylvania

USGS Publications Warehouse

Flippo, H.N.; Behrendt, T.E.

1985-01-01

This report documents a cost-effectiveness study of the stream-gaging program in Pennsylvania. Data uses and funding were identified for 223 continuous-record stream gages operated in 1983; four are planned for discontinuance at the close of water-year 1985; two are suggested for conversion, at the beginning of the 1985 water year, for the collection of only continuous stage records. Two of 11 special-purpose short-term gages are recommended for continuation when the supporting project ends; eight of these gages are to be discontinued and the other will be converted to a partial-record type. Current operation costs for the 212 stations recommended for continued operation is $1,199,000 per year in 1983. The average standard error of estimation for instantaneous streamflow is 15.2%. An overall average standard error of 9.8% could be attained on a budget of $1,271,000, which is 6% greater than the 1983 budget, by adopted cost-effective stream-gaging operations. (USGS)

Scale factor gage for fiber optics inspection device

NASA Technical Reports Server (NTRS)

Mcmahon, W.; Sugg, F. E.

1971-01-01

Flexible wire device, fastened along outside of fiber bundle from viewing portion to tip, positions calibrated adjustable gage in field of view. Scale factor is determined from known magnification characteristics of fiber optics system or from graduations on gage tip.

Cost effectiveness of the stream-gaging program in North Dakota

USGS Publications Warehouse

Ryan, Gerald L.

1989-01-01

This report documents results of a cost-effectiveness study of the stream-gaging program In North Dakota. It is part of a nationwide evaluation of the stream-gaging program of the U.S. Geological Survey.One phase of evaluating cost effectiveness is to identify less costly alternative methods of simulating streamflow records. Statistical or hydro logic flow-routing methods were used as alternative methods to simulate streamflow records for 21 combinations of gaging stations from the 94-gaging-station network. Accuracy of the alternative methods was sufficient to consider discontinuing only one gaging station.Operation of the gaging-station network was evaluated by using associated uncertainty in streamflow records. The evaluation was limited to the nonwinter operation of 29 gaging stations in eastern North Dakota. The current (1987) travel routes and measurement frequencies require a budget of about $248/000 and result in an average equivalent Gaussian spread in streamflow records of 16.5 percent. Changes in routes and measurement frequencies optimally could reduce the average equivalent Gaussian spread to 14.7 percent.Budgets evaluated ranged from $235,000 to $400,000. A $235,000 budget would increase the optimal average equivalent Gaussian spread from 14.7 to 20.4 percent, and a $400,000 budget could decrease it to 5.8 percent.

27. Detail view of mechanical float gages used to monitor ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

27. Detail view of mechanical float gages used to monitor level of water in the filtration bed reservoir. Gage on left measures water head, gage on right monitors filtration rate. - Lake Whitney Water Filtration Plant, Filtration Plant, South side of Armory Street between Edgehill Road & Whitney Avenue, Hamden, New Haven County, CT

Development and characterization of PdCr temperature-compensated wire resistance strain gage

NASA Technical Reports Server (NTRS)

Lei, Jih-Fen

1989-01-01

A temperature-compensated resistance static strain gage with potential to be used to 600 C was recently developed. Gages were fabricated from specially developed palladium-13 w/o chromium (Pd-13Cr) wire and platinum (Pt) compensator. When bonded to high temperature Hastelloy X, the apparent strain from room temperature to 600 C was within 400 microstrain for gages with no preheat treatment and within 3500 microstrain for gages with 16 hours prestabilization at 640 C. The apparent strain versus temperature relationship of stabilized PdCr gages were repeatable with the reproducibility within 100 microstrain during three thermal cycles to 600 C and an 11 hours soak at 600 C. The gage fabrication, construction and installation is described. Also, the coating system used for this compensated resistance strain gage is explained. The electrical properties of the strain sensing element and main characteristics of the compensated gage including apparent strain, drift and reproducibility are discussed.

Strain gage selection in loads equations using a genetic algorithm

NASA Technical Reports Server (NTRS)

1994-01-01

Traditionally, structural loads are measured using strain gages. A loads calibration test must be done before loads can be accurately measured. In one measurement method, a series of point loads is applied to the structure, and loads equations are derived via the least squares curve fitting algorithm using the strain gage responses to the applied point loads. However, many research structures are highly instrumented with strain gages, and the number and selection of gages used in a loads equation can be problematic. This paper presents an improved technique using a genetic algorithm to choose the strain gages used in the loads equations. Also presented are a comparison of the genetic algorithm performance with the current T-value technique and a variant known as the Best Step-down technique. Examples are shown using aerospace vehicle wings of high and low aspect ratio. In addition, a significant limitation in the current methods is revealed. The genetic algorithm arrived at a comparable or superior set of gages with significantly less human effort, and could be applied in instances when the current methods could not.

High Temperature Capacitive Strain Gage

NASA Technical Reports Server (NTRS)

Wnuk, Stephen P., Jr.; Wnuk, Stephen P., III; Wnuk, V. P.

1990-01-01

Capacitive strain gages designed for measurements in wind tunnels to 2000 F were built and evaluated. Two design approaches were followed. One approach was based on fixed capacitor plates with a movable ground plane inserted between the plates to effect differential capacitive output with strain. The second approach was based on movable capacitor plates suspended between sapphire bearings, housed in a rugged body, and arranged to operate as a differential capacitor. A sapphire bearing gage (1/4 in. diameter x 1 in. in size) was built with a range of 50,000 and a resolution of 200 microstrain. Apparent strain on Rene' 41 was less than + or - 1000 microstrain from room temperature to 2000 F. Three gage models were built from the Ground Plane Differential concept. The first was 1/4 in. square by 1/32 in. high and useable to 700 F. The second was 1/2 in. square by 1/16 in. high and useable to 1440 F. The third, also 1/2 in. square by 1/16 in. high was expected to operate in the 1600 to 2000 F range, but was not tested because time and funding ended.

High temperature capacitive strain gage

NASA Astrophysics Data System (ADS)

Wnuk, Stephen P., Jr.; Wnuk, Stephen P., III; Wnuk, V. P.

1990-01-01

Capacitive strain gages designed for measurements in wind tunnels to 2000 F were built and evaluated. Two design approaches were followed. One approach was based on fixed capacitor plates with a movable ground plane inserted between the plates to effect differential capacitive output with strain. The second approach was based on movable capacitor plates suspended between sapphire bearings, housed in a rugged body, and arranged to operate as a differential capacitor. A sapphire bearing gage (1/4 in. diameter x 1 in. in size) was built with a range of 50,000 and a resolution of 200 microstrain. Apparent strain on Rene' 41 was less than + or - 1000 microstrain from room temperature to 2000 F. Three gage models were built from the Ground Plane Differential concept. The first was 1/4 in. square by 1/32 in. high and useable to 700 F. The second was 1/2 in. square by 1/16 in. high and useable to 1440 F. The third, also 1/2 in. square by 1/16 in. high was expected to operate in the 1600 to 2000 F range, but was not tested because time and funding ended.

Gage monitors quality of cross-wire resistance welds

NASA Technical Reports Server (NTRS)

Etzel, J.; Piltch, A.

1968-01-01

Gage nondestructively monitors the quality of cross-wire resistance welds during the welding operation. The gage gives a dial indication of the relative embedment of the cross wires during the actual welding operation. A direct relationship exists between the depth of embedment and both weld strength and consistency.

Empirical Observations on the Sensitivity of Hot Cathode Ionization Type Vacuum Gages

NASA Technical Reports Server (NTRS)

Summers, R. L.

1969-01-01

A study of empirical methods of predicting tile relative sensitivities of hot cathode ionization gages is presented. Using previously published gage sensitivities, several rules for predicting relative sensitivity are tested. The relative sensitivity to different gases is shown to be invariant with gage type, in the linear range of gage operation. The total ionization cross section, molecular and molar polarizability, and refractive index are demonstrated to be useful parameters for predicting relative gage sensitivity. Using data from the literature, the probable error of predictions of relative gage sensitivity based on these molecular properties is found to be about 10 percent. A comprehensive table of predicted relative sensitivities, based on empirical methods, is presented.

Development of strain gages for use to 1311 K (1900 F)

NASA Technical Reports Server (NTRS)

Lemcoe, M. M.

1974-01-01

A high temperature electric resistance strain gage system was developed and evaluated to 1366 K (2000 F) for periods of at least one hour. Wire fabricated from a special high temperature strain gage alloy (BCL-3), was used to fabricate the gages. Various joining techniques (NASA butt welding, pulse arc, plasma needle arc, and dc parallel gap welding) were investigated for joining gage filaments to each other, gage filaments to lead-tab ribbons, and lead-tab ribbons to lead wires. The effectiveness of a clad-wire concept as a means of minimizing apparent strain of BCL-3 strain gages was investigated by sputtering platinum coatings of varying thicknesses on wire samples and establishing the optimum coating thickness--in terms of minimum resistivity changes with temperature. Finally, the moisture-proofing effectiveness of barrier coatings subjected to elevated temperatures was studied, and one commercial barrier coating (BLH Barrier H Waterproofing) was evaluated.

Development of gage widening projection parameter for the deployable gage restraint measurement system

DOT National Transportation Integrated Search

2006-10-01

Recent gage restraint measurement system (GRMS) developments include the redesign of GRMS vehicles to conduct testing from a deployable axle instead of using freight truck mounted axle and GRMS on hi-rail vehicles. This new test configuration results...

Theory and Practice of Shear/Stress Strain Gage Hygrometry

NASA Technical Reports Server (NTRS)

Shams, Qamar A.; Fenner, Ralph L.

2006-01-01

Mechanical hygrometry has progressed during the last several decades from crude hygroscopes to state-of-the art strain-gage sensors. The strain-gage devices vary from different metallic beams to strain-gage sensors using cellulose crystallite elements, held in full shear restraint. This old technique is still in use but several companies are now actively pursuing development of MEMS miniaturized humidity sensors. These new sensors use polyimide thin film for water vapor adsorption and desorption. This paper will provide overview about modern humidity sensors.

Field manual for the collection of Navajo Nation streamflow-gage data

USGS Publications Warehouse

Hart, Robert J.; Fisk, Gregory G.

2014-01-01

The Field Manual for the Collection of Navajo Nation Streamflow-Gage Data (Navajo Field Manual) is based on established (standard) U.S. Geological Survey streamflow-gaging methods and provides guidelines specifically designed for the Navajo Department of Water Resources personnel who establish and maintain streamflow gages. The Navajo Field Manual addresses field visits, including essential field equipment and the selection of and routine visits to streamflow-gaging stations, examines surveying methods for determining peak flows (indirect measurements), discusses safety considerations, and defines basic terms.

Cost effectiveness of the stream-gaging program in Nevada

USGS Publications Warehouse

Arteaga, F.E.

1990-01-01

The stream-gaging network in Nevada was evaluated as part of a nationwide effort by the U.S. Geological Survey to define and document the most cost-effective means of furnishing streamflow information. Specifically, the study dealt with 79 streamflow gages and 2 canal-flow gages that were under the direct operation of Nevada personnel as of 1983. Cost-effective allocations of resources, including budget and operational criteria, were studied using statistical procedures known as Kalman-filtering techniques. The possibility of developing streamflow data at ungaged sites was evaluated using flow-routing and statistical regression analyses. Neither of these methods provided sufficiently accurate results to warrant their use in place of stream gaging. The 81 gaging stations were being operated in 1983 with a budget of $465,500. As a result of this study, all existing stations were determined to be necessary components of the program for the foreseeable future. At the 1983 funding level, the average standard error of streamflow records was nearly 28%. This same overall level of accuracy could have been maintained with a budget of approximately $445,000 if the funds were redistributed more equitably among the gages. The maximum budget analyzed, $1,164 ,000 would have resulted in an average standard error of 11%. The study indicates that a major source of error is lost data. If perfectly operating equipment were available, the standard error for the 1983 program and budget could have been reduced to 21%. (Thacker-USGS, WRD)

Effect of gage size on the measurement of local heat flux. [formulas for determining gage averaging errors

NASA Technical Reports Server (NTRS)

Baumeister, K. J.; Papell, S. S.

1973-01-01

General formulas are derived for determining gage averaging errors of strip-type heat flux meters used in the measurement of one-dimensional heat flux distributions. In addition, a correction procedure is presented which allows a better estimate for the true value of the local heat flux. As an example of the technique, the formulas are applied to the cases of heat transfer to air slot jets impinging on flat and concave surfaces. It is shown that for many practical problems, the use of very small heat flux gages is often unnecessary.

High temperature strain measurement with a resistance strain gage

NASA Technical Reports Server (NTRS)

Lei, Jih-Fen; Fichtel, ED; Mcdaniel, Amos

1993-01-01

A PdCr based electrical resistance strain gage was demonstrated in the laboratory to be a viable sensor candidate for static strain measurement at high temperatures. However, difficulties were encountered while transferring the sensor to field applications. This paper is therefore prepared for recognition and resolution of the problems likely to be encountered with PdCr strain gages in field applications. Errors caused by the measurement system, installation technique and lead wire attachment are discussed. The limitations and some considerations related to the temperature compensation technique used for this gage are also addressed.

Cost effectiveness of the stream-gaging program in northeastern California

USGS Publications Warehouse

Hoffard, S.H.; Pearce, V.F.; Tasker, Gary D.; Doyle, W.H.

1984-01-01

Results are documented of a study of the cost effectiveness of the stream-gaging program in northeastern California. Data uses and funding sources were identified for the 127 continuous stream gages currently being operated in the study area. One stream gage was found to have insufficient data use to warrant cooperative Federal funding. Flow-routing and multiple-regression models were used to simulate flows at selected gaging stations. The models may be sufficiently accurate to replace two of the stations. The average standard error of estimate of streamflow records is 12.9 percent. This overall level of accuracy could be reduced to 12.0 percent using computer-recommended service routes and visit frequencies. (USGS)

Cost-effectiveness of the stream-gaging program in the Hawaii District

USGS Publications Warehouse

Matsuoka, I.; Lee, R.; Thomas, W.O.

1985-01-01

This project documents the results of a study of the cost-effectiveness of the stream-gaging program in the Hawaii District. The stream gages in the District were divided into two groups, the State of Hawaii and the Other Pacific Areas. Data uses and funding sources were identified for the 124 continuous stream gages currently being operated in the Hawaii District with a budget of $570,620. All the stream-gages were identified as having sufficient reason to continue their operation and they should be maintained in the program for the foreseeable future. (USGS)

Practical approaches for application of resistance type strain gages on high temperature composites

NASA Technical Reports Server (NTRS)

Moore, Thomas C., Sr.

1991-01-01

Four major areas of interest with respect to utilizing strain gages on carbon/carbon (with SiC surfaces) and titanium matrix composites are addressed. Strain gage and adhesive combinations on carbon/carbon (C/C) at temperatures from minus 190 C to to 540 C, half-bridge gaging for reducing apparent strain on C/C using Poisson's ratio and bending configurations, a review of the 'field installation' techniques developed for gaging a C/C hypersonic generic elevon, and results of initial strain gaging efforts on titanium matrix composites are discussed. Current research in developing techniques for increasing the maximum temperature for strain gages on carbon/carbon are reviewed.

Evaluation of Pd-Cr Wires for Strain Gage Application

NASA Technical Reports Server (NTRS)

Lei, Jih-Fen; Greer, L. C., III; Oberle, L. G.

1995-01-01

A newly developed alloy, palladium-13 weight percent chromium (Pd13Cr), was identified by United Technologies Research Center under a NASA contract to be the best material for high temperature strain gage applications. An electrical resistance strain gage that can provide accurate static strain measurement to a temperature higher than that of a commercially available gage is urgently needed in aerospace and aeronautics research. A strain gage made of a 25.4 micron (1 mil) diameter Pd13Cr wire has been recently demonstrated to be usable for static strain measurements to 800 C. This compares to the 400 C temperature limit of the commercially available strain gages. The performance of the Pd-Cr gage, however, strongly depends on the quality of the Pd13Cr wire. Four batches of Pd-Cr wires purchased from three different manufacturers were therefore evaluated to determine the best source of the wire for strain gage applications. The three suppliers were Precious Metal Institute in China, Sigmund Cohn Co., and G & S Titanium, Inc. in the United States. Two batches of wires obtained from Previous Metal Institute in 1987 and 1992, respectively are referred to herein as China87 and China92 wires. The mechanical, chemical and electrical properties of these wires, both as-received and after high temperature exposures at 800 C for 50 hours were analyzed. The elastic modulus and the failure strength of the wires were evaluated using a tensile test machine equipped with a laser speckle strain measurement system. The chemical and microstructural properties of the wires were inspected using a plasma atomic emission spectrometer and a scanning electron microscope (SEM) equipped with an energy dispersive X-ray spectroscope (EDS). The electrical stability and repeatability of the wires were determined by measuring the electrical resistance of the wires during three thermal cycles to 1000 C and a ten-hour soak at 1000 C. As a result of this study, the wire which has the highest

Influence of Primary Gage Sensitivities on the Convergence of Balance Load Iterations

NASA Technical Reports Server (NTRS)

Ulbrich, Norbert Manfred

2012-01-01

The connection between the convergence of wind tunnel balance load iterations and the existence of the primary gage sensitivities of a balance is discussed. First, basic elements of two load iteration equations that the iterative method uses in combination with results of a calibration data analysis for the prediction of balance loads are reviewed. Then, the connection between the primary gage sensitivities, the load format, the gage output format, and the convergence characteristics of the load iteration equation choices is investigated. A new criterion is also introduced that may be used to objectively determine if the primary gage sensitivity of a balance gage exists. Then, it is shown that both load iteration equations will converge as long as a suitable regression model is used for the analysis of the balance calibration data, the combined influence of non linear terms of the regression model is very small, and the primary gage sensitivities of all balance gages exist. The last requirement is fulfilled, e.g., if force balance calibration data is analyzed in force balance format. Finally, it is demonstrated that only one of the two load iteration equation choices, i.e., the iteration equation used by the primary load iteration method, converges if one or more primary gage sensitivities are missing. This situation may occur, e.g., if force balance calibration data is analyzed in direct read format using the original gage outputs. Data from the calibration of a six component force balance is used to illustrate the connection between the convergence of the load iteration equation choices and the existence of the primary gage sensitivities.

Optical ablation/temperature gage (COTA)

NASA Astrophysics Data System (ADS)

Cassaing, J.; Balageas, D.

ONERA has ground and flight tested for heat-shield recession a novel technique, different from current radiation and acoustic measurement methods. It uses a combined ablation/temperature gage that views the radiation optically from a cavity embedded within the heat shield. Flight measurements, both of temperature and of passage of the ablation front, are compared with data generated by a predictive numerical code. The ablation and heat diffusion into the instrumented ablator can be simulated numerically to evaluate accurately the errors due to the presence of the gage. This technology was established in 1978 and finally adopted after ground tests in arc heater facilities. After four years of flight evaluations, it is possible to evaluate and criticize the sensor reliability.

49 CFR 213.143 - Frog guard rails and guard faces; gage.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 49 Transportation 4 2013-10-01 2013-10-01 false Frog guard rails and guard faces; gage. 213.143... and guard faces; gage. The guard check and guard face gages in frogs shall be within the limits... frog to the guard line 1 of its guard rail or guarding face, measured across the track at right angles...

49 CFR 213.143 - Frog guard rails and guard faces; gage.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 49 Transportation 4 2011-10-01 2011-10-01 false Frog guard rails and guard faces; gage. 213.143... and guard faces; gage. The guard check and guard face gages in frogs shall be within the limits... frog to the guard line 1 of its guard rail or guarding face, measured across the track at right angles...

49 CFR 213.143 - Frog guard rails and guard faces; gage.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 4 2010-10-01 2010-10-01 false Frog guard rails and guard faces; gage. 213.143... and guard faces; gage. The guard check and guard face gages in frogs shall be within the limits... frog to the guard line 1 of its guard rail or guarding face, measured across the track at right angles...

49 CFR 213.143 - Frog guard rails and guard faces; gage.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 4 2014-10-01 2014-10-01 false Frog guard rails and guard faces; gage. 213.143... and guard faces; gage. The guard check and guard face gages in frogs shall be within the limits... frog to the guard line 1 of its guard rail or guarding face, measured across the track at right angles...

49 CFR 213.143 - Frog guard rails and guard faces; gage.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 49 Transportation 4 2012-10-01 2012-10-01 false Frog guard rails and guard faces; gage. 213.143... and guard faces; gage. The guard check and guard face gages in frogs shall be within the limits... frog to the guard line 1 of its guard rail or guarding face, measured across the track at right angles...

Palladium-chromium static strain gage for high temperature propulsion systems

NASA Technical Reports Server (NTRS)

Lei, Jih-Fen

1991-01-01

The present electrical strain gage for high temperature static strain measurements is in its fine-wire and thin-film forms designed to be temperature-compensated on any substrate material. The gage element is of Pd-Cr alloy, while the compensator is of Pt. Because the thermally-induced apparent strain of this compensated wire strain gage is sufficiently small, with good reproducibility between thermal cycles to 800 C, output figures can be corrected within a reasonable margin of error.

Gage for micromachining system

DOEpatents

Miller, Donald M.

1979-02-27

A gage for measuring the contour of the surface of an element of a micromachining tool system and of a work piece machined by the micromachining tool system. The gage comprises a glass plate containing two electrical contacts and supporting a steel ball resting against the contacts. As the element or workpiece is moved against the steel ball, the very slight contact pressure causes an extremely small movement of the steel ball which breaks the electrical circuit between the two contacts. The contour information is supplied to a dedicated computer controlling the micromachining tool so that the computer knows the contour of the element and the work piece to an accuracy of .+-. 25 nm. The micromachining tool system with X- and omega-axes is used to machine spherical, aspherical, and irregular surfaces with a maximum contour error of 100 nanometers (nm) and surface waviness of no more than 0.8 nm RMS.

Comparison of thin-film resistance heat-transfer gages with thin-skin transient calorimeter gages in conventional hypersonic wind tunnels

NASA Technical Reports Server (NTRS)

Miller, C. G., III

1981-01-01

Thin film gages deposited at the stagnation region of small (8.1-mm-diameter) hemispheres and gages mounted flush with the surface of a sharp-leading-edge flat plate were tested in the Langley continuous-flow hypersonic tunnel and in the Langley hypersonic CF4 tunnel. Two substrate materials were tested, quartz and a machinable glass-ceramic. Small hemispheres were also tested utilizing the thin-skin transient calorimeter technique usually employed in conventional tunnels. One transient calorimeter model was a thin shell of stainless steel, and the other was a thin-skin insert of stainless steel mounted into a hemisphere fabricated from a machinable-glass-ceramic. Measured heat-transfer rates from the various hemispheres were compared with one another and with predicted rates. The results demonstrate the feasibility and advantages of using-film resistance heat-transfer gages in conventional hypersonic wind tunnels over a wide range of conditions.

Cost-effectiveness of the U.S. Geological Survey stream-gaging program in Indiana

USGS Publications Warehouse

Stewart, J.A.; Miller, R.L.; Butch, G.K.

1986-01-01

Analysis of the stream gaging program in Indiana was divided into three phases. The first phase involved collecting information concerning the data need and the funding source for each of the 173 surface water stations in Indiana. The second phase used alternate methods to produce streamflow records at selected sites. Statistical models were used to generate stream flow data for three gaging stations. In addition, flow routing models were used at two of the sites. Daily discharges produced from models did not meet the established accuracy criteria and, therefore, these methods should not replace stream gaging procedures at those gaging stations. The third phase of the study determined the uncertainty of the rating and the error at individual gaging stations, and optimized travel routes and frequency of visits to gaging stations. The annual budget, in 1983 dollars, for operating the stream gaging program in Indiana is $823,000. The average standard error of instantaneous discharge for all continuous record gaging stations is 25.3%. A budget of $800,000 could maintain this level of accuracy if stream gaging stations were visited according to phase III results. A minimum budget of $790,000 is required to operate the gaging network. At this budget, the average standard error of instantaneous discharge would be 27.7%. A maximum budget of $1 ,000,000 was simulated in the analysis and the average standard error of instantaneous discharge was reduced to 16.8%. (Author 's abstract)

Cost effectiveness of the US Geological Survey's stream-gaging program in New York

USGS Publications Warehouse

Wolcott, S.W.; Gannon, W.B.; Johnston, W.H.

1986-01-01

The U.S. Geological Survey conducted a 5-year nationwide analysis to define and document the most cost effective means of obtaining streamflow data. This report describes the stream gaging network in New York and documents the cost effectiveness of its operation; it also identifies data uses and funding sources for the 174 continuous-record stream gages currently operated (1983). Those gages as well as 189 crest-stage, stage-only, and groundwater gages are operated with a budget of $1.068 million. One gaging station was identified as having insufficient reason for continuous operation and was converted to a crest-stage gage. Current operation of the 363-station program requires a budget of $1.068 million/yr. The average standard error of estimation of continuous streamflow data is 13.4%. Results indicate that this degree of accuracy could be maintained with a budget of approximately $1.006 million if the gaging resources were redistributed among the gages. The average standard error for 174 stations was calculated for five hypothetical budgets. A minimum budget of $970,000 would be needed to operated the 363-gage program; a budget less than this does not permit proper servicing and maintenance of the gages and recorders. Under the restrictions of a minimum budget, the average standard error would be 16.0%. The maximum budget analyzed was $1.2 million, which would decrease the average standard error to 9.4%. (Author 's abstract)

High temperature strain gage technology for gas turbine engines

NASA Technical Reports Server (NTRS)

Fichtel, Edward J.; Mcdaniel, Amos D.

1994-01-01

This report summarizes the results of a six month study that addressed specific issues to transfer the Pd-13Cr static strain sensor to a gas turbine engine environment. The application issues that were addressed include: (1) evaluation of a miniature, variable potentiometer for use as the ballast resistor, in conjunction with a conventional strain gage signal conditioning unit; (2) evaluation of a metal sheathed, platinum conductor leadwire assembly for use with the three-wire sensor; and (3) subjecting the sensor to dynamic strain cyclic testing to determine fatigue characteristics. Results indicate a useful static strain gage system at all temperature levels up to 1350 F. The fatigue characteristics also appear to be very promising, indicating a potential use in dynamic strain measurement applications. The procedure, set-up, and data for all tests are presented in this report. This report also discusses the specific strain gage installation technique for the Pd-13Cr gage because of its potential impact on the quality of the output data.

Differences between nipher and slter shielded rain gages at two Colorado deposition monitoring sites

USGS Publications Warehouse

Bigelow, David S.; Denning, A. Scott

1990-01-01

In the last decade the United States and Canada have made significant progress in establishing spatial ad temporal estimates of atmospheric deposition throughout North America. Fundamental to the wet-deposition portion of these estimates is the accurate and precise measurement of precipitation amount. Goodison and others (I-3) have reported on a new type of shielded snow gage known as the Canadian MSC Nipher shielded snow gage. Because this shielded snow gage has been shown to be superior to other precipitation gages for the estimation of snowfall amount, its design was adapted to the Universal Belfort precipitation gage (4), the dominant precipitation gage used at deposition monitoring sites in the United States. Favorable results taken from monitoring sites using this modified Nipher shielded snow gage (3-6) have prompted the U.S. Environmental Protection Agency and the Electric Power Research Institute to adopt the Nipher shielded Belfort gage as a standard piece of equipment in the Acid MODES and Operational Evaluation Network (OEN) monitoring programs and to propose that is be included as a standard snow gage in other North American deposition monitoring programs. This communication details preliminary results from two of nine NADP/NTN deposition monitoring sites selected by the Environmental Protection Agency to compare Nipher shielded Belfort precipitation gage volumes to volumes obtained from the standard Belfort gage used in the NADP/NTN monitoring program.

Application of Temperature Sensitivities During Iterative Strain-Gage Balance Calibration Analysis

NASA Technical Reports Server (NTRS)

Ulbrich, N.

2011-01-01

A new method is discussed that may be used to correct wind tunnel strain-gage balance load predictions for the influence of residual temperature effects at the location of the strain-gages. The method was designed for the iterative analysis technique that is used in the aerospace testing community to predict balance loads from strain-gage outputs during a wind tunnel test. The new method implicitly applies temperature corrections to the gage outputs during the load iteration process. Therefore, it can use uncorrected gage outputs directly as input for the load calculations. The new method is applied in several steps. First, balance calibration data is analyzed in the usual manner assuming that the balance temperature was kept constant during the calibration. Then, the temperature difference relative to the calibration temperature is introduced as a new independent variable for each strain--gage output. Therefore, sensors must exist near the strain--gages so that the required temperature differences can be measured during the wind tunnel test. In addition, the format of the regression coefficient matrix needs to be extended so that it can support the new independent variables. In the next step, the extended regression coefficient matrix of the original calibration data is modified by using the manufacturer specified temperature sensitivity of each strain--gage as the regression coefficient of the corresponding temperature difference variable. Finally, the modified regression coefficient matrix is converted to a data reduction matrix that the iterative analysis technique needs for the calculation of balance loads. Original calibration data and modified check load data of NASA's MC60D balance are used to illustrate the new method.

Development of Displacement Gages Exposed to Solid Rocket Motor Internal Environments

NASA Technical Reports Server (NTRS)

Bolton, D. E.; Cook, D. J.

2003-01-01

The Space Shuttle Reusable Solid Rocket Motor (RSRM) has three non-vented segment-to-segment case field joints. These joints use an interference fit J-joint that is bonded at assembly with a Pressure Sensitive Adhesive (PSA) inboard of redundant O-ring seals. Full-scale motor and sub-scale test article experience has shown that the ability to preclude gas leakage past the J-joint is a function of PSA type, joint moisture from pre-assembly humidity exposure, and the magnitude of joint displacement during motor operation. To more accurately determine the axial displacements at the J-joints, two thermally durable displacement gages (one mechanical and one electrical) were designed and developed. The mechanical displacement gage concept was generated first as a non-electrical, self-contained gage to capture the maximum magnitude of the J-joint motion. When it became feasible, the electrical displacement gage concept was generated second as a real-time linear displacement gage. Both of these gages were refined in development testing that included hot internal solid rocket motor environments and simulated vibration environments. As a result of this gage development effort, joint motions have been measured in static fired RSRM J-joints where intentional venting was produced (Flight Support Motor #8, FSM-8) and nominal non-vented behavior occurred (FSM-9 and FSM-10). This data gives new insight into the nominal characteristics of the three case J-joint positions (forward, center and aft) and characteristics of some case J-joints that became vented during motor operation. The data supports previous structural model predictions. These gages will also be useful in evaluating J-joint motion differences in a five-segment Space Shuttle solid rocket motor.

High temperature static strain measurement with an electrical resistance strain gage

NASA Technical Reports Server (NTRS)

Lei, Jih-Fen

1992-01-01

An electrical resistance strain gage that can supply accurate static strain measurement for NASP application is being developed both in thin film and fine wire forms. This gage is designed to compensate for temperature effects on substrate materials with a wide range of thermal expansion coefficients. Some experimental results of the wire gage tested on one of the NASP structure materials, i.e., titanium matrix composites, are presented.

In-situ shear stress indicator using heated strain gages at the flow boundary

NASA Astrophysics Data System (ADS)

Yeh, Chi-An; Yang, Fuling

2011-11-01

This work borrows the concept of hot-wire anemometry and sketch a technique that uses local heat transfer to infer the flow field and the corresponding stress. Conventional strain gages were mounted at the flow solid boundary as the heat source and acrylic boundary was chosen for its low thermal conductivity ensuring heat accumulation when a gage is energized. The gage would now work in slightly overheated state and its self-heating leads to an additional thermal strain. When exposed to a flow field, heat is brought away by local forced convection, resulting in deviations in gage signal from that developed in quiescent liquid. We have developed a facility to achieve synchronous gage measurements at different locations on a solid boundary. Three steady flow motions were considered: circular Couette flow, rectilinear uniform flow, and rectilinear oscillating flow. Preliminary tests show the gage reading does respond to the imposed flow through thermal effects and greater deviation was measured in flows of higher shear strain rates. The correlation between the gage signals and the imposed flow field is further examined by theoretical analysis. We also introduced a second solid boundary to the vicinity of the gage in the two rectilinear flows. The gage readings demonstrate rises in its magnitudes indicating wall amplification effect on the local shear strain, agreeing to the drag augmentation by a second solid boundary reported in many multiphase flow literatures.

Expression Differentiation Is Constrained to Low-Expression Proteins over Ecological Timescales

PubMed Central

Margres, Mark J.; Wray, Kenneth P.; Seavy, Margaret; McGivern, James J.; Herrera, Nathanael D.; Rokyta, Darin R.

2016-01-01

Protein expression level is one of the strongest predictors of protein sequence evolutionary rate, with high-expression protein sequences evolving at slower rates than low-expression protein sequences largely because of constraints on protein folding and function. Expression evolutionary rates also have been shown to be negatively correlated with expression level across human and mouse orthologs over relatively long divergence times (i.e., ∼100 million years). Long-term evolutionary patterns, however, often cannot be extrapolated to microevolutionary processes (and vice versa), and whether this relationship holds for traits evolving under directional selection within a single species over ecological timescales (i.e., <5000 years) is unknown and not necessarily expected. Expression is a metabolically costly process, and the expression level of a particular protein is predicted to be a tradeoff between the benefit of its function and the costs of its expression. Selection should drive the expression level of all proteins close to values that maximize fitness, particularly for high-expression proteins because of the increased energetic cost of production. Therefore, stabilizing selection may reduce the amount of standing expression variation for high-expression proteins, and in combination with physiological constraints that may place an upper bound on the range of beneficial expression variation, these constraints could severely limit the availability of beneficial expression variants. To determine whether rapid-expression evolution was restricted to low-expression proteins owing to these constraints on highly expressed proteins over ecological timescales, we compared venom protein expression levels across mainland and island populations for three species of pit vipers. We detected significant differentiation in protein expression levels in two of the three species and found that rapid-expression differentiation was restricted to low-expression proteins. Our

Estimating Low-Flow Frequency Statistics and Hydrologic Analysis of Selected Streamflow-Gaging Stations, Nooksack River Basin, Northwestern Washington and Canada

USGS Publications Warehouse

Curran, Christopher A.; Olsen, Theresa D.

2009-01-01

Low-flow frequency statistics were computed at 17 continuous-record streamflow-gaging stations and 8 miscellaneous measurement sites in and near the Nooksack River basin in northwestern Washington and Canada, including the 1, 3, 7, 15, 30, and 60 consecutive-day low flows with recurrence intervals of 2 and 10 years. Using these low-flow statistics, 12 regional regression equations were developed for estimating the same low-flow statistics at ungaged sites in the Nooksack River basin using a weighted-least-squares method. Adjusted R2 (coefficient of determination) values for the equations ranged from 0.79 to 0.93 and the root-mean-squared error (RMSE) expressed as a percentage ranged from 77 to 560 percent. Streamflow records from six gaging stations located in mountain-stream or lowland-stream subbasins of the Nooksack River basin were analyzed to determine if any of the gaging stations could be removed from the network without significant loss of information. Using methods of hydrograph comparison, daily-value correlation, variable space, and flow-duration ratios, and other factors relating to individual subbasins, the six gaging stations were prioritized from most to least important as follows: Skookum Creek (12209490), Anderson Creek (12210900), Warm Creek (12207750), Fishtrap Creek (12212050), Racehorse Creek (12206900), and Clearwater Creek (12207850). The optimum streamflow-gaging station network would contain all gaging stations except Clearwater Creek, and the minimum network would include Skookum Creek and Anderson Creek.

Embedded gage impact study

NASA Technical Reports Server (NTRS)

Schramm, S. W.; Daniel, I. M.

1984-01-01

Impact damage in graphite/epoxy laminates was characterized and transient strain history during impact was correlated. The material investigated was AS-4/3501-6 graphite/epoxy. Eight-ply and sixteen-ply quasi-isotropic laminates of 45/0/-45/90 sub s and 45/0/-45/90 sub 2s layups were fabricated with strain gages embedded between plies during the strain gages and leads from the highly conductive graphite fibers. The specimens were circular plates 12.7 cm (5 in.) in diameter and clamped along their circumference. The specimens were impacted with a 185 gm impactor, dropped from heights of 1.20 m and 1.65 m. An accelerometer was attached to the back surface of the specimen opposite the impact point and was used to trigger the recording instrumentation. The transient strain data were recorded with an eight channel waveform digitizer capable of sampling data at 0.5 microsec intervals. The data were stored, processed, and plotted by means of a microcomputer. Transient strain data were correlated with results from ultrasonic inspection of the specimens.

Graphical correlation of gaging-station records

USGS Publications Warehouse

Searcy, James K.

1960-01-01

A gaging-station record is a sample of the rate of flow of a stream at a given site. This sample can be used to estimate the magnitude and distribution of future flows if the record is long enough to be representative of the long-term flow of the stream. The reliability of a short-term record for estimating future flow characteristics can be improved through correlation with a long-term record. Correlation can be either numerical or graphical, but graphical correlation of gaging-station records has several advantages. The graphical correlation method is described in a step-by-step procedure with an illustrative problem of simple correlation, illustrative problems of three examples of multiple correlation--removing seasonal effect--and two examples of correlation of one record with two other records. Except in the problem on removal of seasonal effect, the same group of stations is used in the illustrative problems. The purpose of the problems is to illustrate the method--not to show the improvement that can result from multiple correlation as compared with simple correlation. Hydrologic factors determine whether a usable relation exists between gaging-station records. Statistics is only a tool for evaluating and using an existing relation, and the investigator must be guided by a knowledge of hydrology.

Cost effectiveness of the stream-gaging program in South Carolina

USGS Publications Warehouse

Barker, A.C.; Wright, B.C.; Bennett, C.S.

1985-01-01

The cost effectiveness of the stream-gaging program in South Carolina was documented for the 1983 water yr. Data uses and funding sources were identified for the 76 continuous stream gages currently being operated in South Carolina. The budget of $422,200 for collecting and analyzing streamflow data also includes the cost of operating stage-only and crest-stage stations. The streamflow records for one stream gage can be determined by alternate, less costly methods, and should be discontinued. The remaining 75 stations should be maintained in the program for the foreseeable future. The current policy for the operation of the 75 stations including the crest-stage and stage-only stations would require a budget of $417,200/yr. The average standard error of estimation of streamflow records is 16.9% for the present budget with missing record included. However, the standard error of estimation would decrease to 8.5% if complete streamflow records could be obtained. It was shown that the average standard error of estimation of 16.9% could be obtained at the 75 sites with a budget of approximately $395,000 if the gaging resources were redistributed among the gages. A minimum budget of $383,500 is required to operate the program; a budget less than this does not permit proper service and maintenance of the gages and recorders. At the minimum budget, the average standard error is 18.6%. The maximum budget analyzed was $850,000, which resulted in an average standard error of 7.6 %. (Author 's abstract)

An electrical strain gage for the tensile testing of paper

Treesearch

Douglas M. Jewett

1963-01-01

A direct-reading strain gage has been developed at the U.S. Forest Products Laboratory that provides rapid and accurate measurement of the stress - strain properties of paper. The gage, which employs a differential transformer, is particularly suited to servo-operated x-y recorders.

Single-strain-gage force/stiffness buckling prediction techniques on a hat-stiffened panel

NASA Technical Reports Server (NTRS)

Hudson, Larry D.; Thompson, Randolph C.

1991-01-01

Predicting the buckling characteristics of a test panel is necessary to ensure panel integrity during a test program. A single-strain-gage buckling prediction method was developed on a hat-stiffened, monolithic titanium buckling panel. The method is an adaptation of the original force/stiffness method which requires back-to-back gages. The single-gage method was developed because the test panel did not have back-to-back gages. The method was used to predict buckling loads and temperatures under various heating and loading conditions. The results correlated well with a finite element buckling analysis. The single-gage force/stiffness method was a valid real-time and post-test buckling prediction technique.

Cost effectiveness of stream-gaging program in Michigan

USGS Publications Warehouse

Holtschlag, D.J.

1985-01-01

This report documents the results of a study of the cost effectiveness of the stream-gaging program in Michigan. Data uses and funding sources were identified for the 129 continuous gaging stations being operated in Michigan as of 1984. One gaging station was identified as having insufficient reason to continue its operation. Several stations were identified for reactivation, should funds become available, because of insufficiencies in the data network. Alternative methods of developing streamflow information based on routing and regression analyses were investigated for 10 stations. However, no station records were reproduced with sufficient accuracy to replace conventional gaging practices. A cost-effectiveness analysis of the data-collection procedure for the ice-free season was conducted using a Kalman-filter analysis. To define missing-record characteristics, cross-correlation coefficients and coefficients of variation were computed at stations on the basis of daily mean discharge. Discharge-measurement data were used to describe the gage/discharge rating stability at each station. The results of the cost-effectiveness analysis for a 9-month ice-free season show that the current policy of visiting most stations on a fixed servicing schedule once every 6 weeks results in an average standard error of 12.1 percent for the current $718,100 budget. By adopting a flexible servicing schedule, the average standard error could be reduced to 11.1 percent. Alternatively, the budget could be reduced to $700,200 while maintaining the current level of accuracy. A minimum budget of $680,200 is needed to operate the 129-gaging-station program; a budget less than this would not permit proper service and maintenance of stations. At the minimum budget, the average standard error would be 14.4 percent. A budget of $789,900 (the maximum analyzed) would result in a decrease in the average standard error to 9.07 percent. Owing to continual changes in the composition of the network

Superlattice strain gage

DOEpatents

Noel, B.W.; Smith, D.L.; Sinha, D.N.

1988-06-28

A strain gage comprising a strained-layer superlattice crystal exhibiting piezoelectric properties is described. A substrate upon which such a strained-layer superlattice crystal has been deposited is attached to an element to be monitored for strain. A light source is focused on the superlattice crystal and the light reflected from, passed through, or emitted from the crystal is gathered and compared with previously obtained optical property data to determine the strain in the element. 8 figs.

Expression of proteins in Escherichia coli as fusions with maltose-binding protein to rescue non-expressed targets in a high-throughput protein-expression and purification pipeline

PubMed Central

Hewitt, Stephen N.; Choi, Ryan; Kelley, Angela; Crowther, Gregory J.; Napuli, Alberto J.; Van Voorhis, Wesley C.

2011-01-01

Despite recent advances, the expression of heterologous proteins in Escherichia coli for crystallization remains a nontrivial challenge. The present study investigates the efficacy of maltose-binding protein (MBP) fusion as a general strategy for rescuing the expression of target proteins. From a group of sequence-verified clones with undetectable levels of protein expression in an E. coli T7 expression system, 95 clones representing 16 phylogenetically diverse organisms were selected for recloning into a chimeric expression vector with an N-terminal histidine-tagged MBP. PCR-amplified inserts were annealed into an identical ligation-independent cloning region in an MBP-fusion vector and were analyzed for expression and solubility by high-throughput nickel-affinity binding. This approach yielded detectable expression of 72% of the clones; soluble expression was visible in 62%. However, the solubility of most proteins was marginal to poor upon cleavage of the MBP tag. This study offers large-scale evidence that MBP can improve the soluble expression of previously non-expressing proteins from a variety of eukaryotic and prokaryotic organisms. While the behavior of the cleaved proteins was disappointing, further refinements in MBP tagging may permit the more widespread use of MBP-fusion proteins in crystallographic studies. PMID:21904041

Recommended Strain Gage Application Procedures for Various Langley Research Center Balances and Test Articles

NASA Technical Reports Server (NTRS)

Moore, Thomas C., Sr.

1997-01-01

The NASA Langley Research Center uses more than 10000 strain gages per year in supporting its various research programs. The character of the testing at LaRC is such that the types of strain gage installations, the materials they are applied to, and the test environments encountered, require many varied approaches for installing strain gages. These installations must be accomplished in the most technically discerning and appropriate manner. This technical memorandum is offered as an assisting guide in helping the strain gage user to determine the appropriate approach for a given strain gage application requirement. Specifically, this document offers detailed recommendations for strain gaging the following: LaRC-Designed balances, LARC custom transducers, certain composite materials and alloys, high-temperature test articles, and selected non-typical or unique materials or test conditions.

High-Temperature Extensometry and PdCr Temperature-Compensated Wire Resistance Strain Gages Compared

NASA Technical Reports Server (NTRS)

1997-01-01

A detailed experimental evaluation is underway at the NASA Lewis Research Center to compare and contrast the performance of the PdCr/Pt dual-element temperature-compensated wire resistance strain gage with that of conventional high-temperature extensometry. The advanced PdCr gage, developed by researchers at Lewis, exhibits desirable properties and a relatively small and repeatable apparent strain to 800 C. This gage represents a significant advance in technology because existing commercial resistance strain gages are not reliable for quasi-static strain measurements above approx. 400 C. Various thermal and mechanical loading spectra are being applied by a high-temperature thermomechanical uniaxial testing system to evaluate the two strain-measurement systems. This is being done not only to compare and contrast the two strain sensors, but also to investigate the applicability of the PdCr strain gage to the coupon-level specimen testing environment typically employed when the high-temperature mechanical behavior of structural materials is characterized. Strain measurement capabilities to 800 C are being investigated with a nickel-base superalloy, Inconel 100 (IN 100), substrate material and application to TMC's is being examined with the model system, SCS-6/Ti-15-3. Furthermore, two gage application techniques are being investigated in the comparison study: namely, flame-sprayed and spot welding. The apparent strain responses of both the weldable and flame-sprayed PdCr wire strain gages were found to be cyclically repeatable on both IN 100 and SCS-6/Ti-15-3 [0]_8. In general, each gage exhibited some uniqueness with respect to apparent strain behavior. Gages mounted on the IN 100 specimens tended to show a repeatable apparent strain within the first few cycles, because the thermal response of IN 100 was stable. This was not the case, however, for the TMC specimens, which typically required several thermal cycles to stabilize the thermal strain response. Thus

The ALE/GAGE/AGAGE Network (DB1001)

DOE Data Explorer

Prinn, Ronald G. [MIT, Center for Global Change Science; Weiss, Ray F. [University of California, San Diego; Scripps Institution of Oceanography; Krummel, Paul B. [CSIRO Oceans and Atmosphere, Cape Grim; O'Doherty, Simon [University of Bristol, Barbados and Mace Head Stations; Fraser, Paul [CSIRO Oceans and Atmosphere; Muhle, Jens [UCSD Scripps Institution of Oceanography; Cape Matatula Station; Reimann, Stefan [Swiss Federal Laboratories for Materials Science and Research (EMPA); Jungfraujoch Station; Vollmer, Martin [Swiss Federal Laboratories for Materials Science and Research (EMPA); Jungfraujoch Station; Simmonds, Peter G. [University of Bristol, Atmospheric Chemistry Research Group; Mace Head Station; Malone, Michela [University of Urbino; Monte Cimone Station; Arduini, Jgor [University of Urbino; Monte Cimone Station; Lunder, Chris [Norwegian Institute for Air Research; Ny Alesund Station; Hermansen, Ove [Norwegian Inst. for Air Research (NILU), Kjeller (Norway); Ny Alesund Station; Schmidbauer, Norbert [Norwegian Inst. for Air Research (NILU), Kjeller (Norway); Global Network; Young, Dickon [University of Bristol; Ragged Point Station; Wang, Hsiang J. (Ray) [Geogia Institute of Technology, School of Earth and Atmospheric Sciences; Global Network; Huang, Jin; Rigby, Matthew [University of Bristol; Global Network; Harth, Chris [UCSD, Scripps Institutioon of Oceanography; Global Network; Salameh, Peter [UCSD, Scripps Institution of Oceanography; Global Network; Spain, Gerard [National University of Ireland; Global Network; Steele, Paul [CSIRO Oceans and Atmosphere; Global Network; Arnold, Tim; Kim, Jooil [UCSD, Scripps Institution of Oceanography; Global Network; Derek, Nada; mitrevski, Blagoj; Langenfelds, Ray

2008-01-01

In the ALE/GAGE/AGAGE global network program, continuous high frequency gas chromatographic measurements of four biogenic/anthropogenic gases (methane, CH4; nitrous oxide, N2O; hydrogen, H; and carbon monoxide, CO) and several anthropogenic gases that contribute to stratospheric ozone destruction and/or to the greenhouse effect have been carried out at five globally distributed sites for several years. The program, which began in 1978, is divided into three parts associated with three changes in instrumentation: the Atmospheric Lifetime Experiment (ALE), which used Hewlett Packard HP5840 gas chromatographs; the Global Atmospheric Gases Experiment (GAGE), which used HP5880 gas chromatographs; and the present Advanced GAGE (AGAGE). AGAGE uses two types of instruments: a gas chromatograph with multiple detectors (GC-MD), and a gas chromatograph with mass spectrometric analysis (GC-MS). Beginning in January 2004, an improved cryogenic preconcentration system (Medusa) replaced the absorption-desorption module in the GC-MS systems at Mace Head and Cape Grim; this provided improved capability to measure a broader range of volatile perfluorocarbons with high global warming potentials. More information may be found at the AGAGE home page: http://agage.eas.gatech.edu/instruments-gcms-medusa.htm.

Hidden Connections between Regression Models of Strain-Gage Balance Calibration Data

NASA Technical Reports Server (NTRS)

Ulbrich, Norbert

2013-01-01

Hidden connections between regression models of wind tunnel strain-gage balance calibration data are investigated. These connections become visible whenever balance calibration data is supplied in its design format and both the Iterative and Non-Iterative Method are used to process the data. First, it is shown how the regression coefficients of the fitted balance loads of a force balance can be approximated by using the corresponding regression coefficients of the fitted strain-gage outputs. Then, data from the manual calibration of the Ames MK40 six-component force balance is chosen to illustrate how estimates of the regression coefficients of the fitted balance loads can be obtained from the regression coefficients of the fitted strain-gage outputs. The study illustrates that load predictions obtained by applying the Iterative or the Non-Iterative Method originate from two related regression solutions of the balance calibration data as long as balance loads are given in the design format of the balance, gage outputs behave highly linear, strict statistical quality metrics are used to assess regression models of the data, and regression model term combinations of the fitted loads and gage outputs can be obtained by a simple variable exchange.

A hot-wire surface gage for skin friction and separation detection measurements

NASA Technical Reports Server (NTRS)

Rubesin, M. W.; Okuno, A. F.; Mateer, G. G.; Brosh, A.

1975-01-01

A heated-element, skin-friction gage employing a very low thermal conductivity support is described. It is shown that the effective dimension of the gage in the stream direction in only 0.06 mm, including the effects of heat conduction in the supporting material. Because of its small size, the calibration of the gage is independent of the kind of boundary-layer flow (whether laminar or turbulent) and is insensitive to pressure gradients. Construction tolerances can be maintained so that a single universal calibration can be applied. Multiple gages, sufficiently closely spaced so as to interfere with each other, are shown to provide accurate determinations of the locations of the points of boundary-layer separation and reattachment.

Demonstration test of burner liner strain measurements using resistance strain gages

NASA Technical Reports Server (NTRS)

Grant, H. P.; Anderson, W. L.

1984-01-01

A demonstration test of burner liner strain measurements using resistance strain gages as well as a feasibility test of an optical speckle technique for strain measurement are presented. The strain gage results are reported. Ten Kanthal A-1 wire strain gages were used for low cycle fatigue strain measurements to 950 K and .002 apparent strain on a JT12D burner can in a high pressure (10 atmospheres) burner test. The procedure for use of the strain gages involved extensive precalibration and postcalibration to correct for cooling rate dependence, drift, and temperature effects. Results were repeatable within + or - .0002 to .0006 strain, with best results during fast decels from 950 K. The results agreed with analytical prediction based on an axisymmetric burner model, and results indicated a non-uniform circumferential distribution of axial strain, suggesting temperature streaking.

Elevated-Temperature Tensile-Testing of Foil-Gage Metals

NASA Technical Reports Server (NTRS)

Blackburn, L. B.; Ellingsworth, J. R.

1986-01-01

Automated system for measuring strain in metal foils at temperatures above 500 degrees F (260 degrees C) uses mechanical extensometer and displacement transducer. System includes counterbalance feature, which eliminates weight contribution of extensometer and reduces grip pressure required for attachment to specimen. Counterbalancing feature overcomes two major difficulties in using extensometers with foil-gage specimens: (1) Weight of extensometer and transducer represents significant fraction of total load applied to specimen and may actually damage it; and (2) grip pressure required for attachment of extensometer to specimens may induce bending stresses in foil-gage materials.

Development of buried wire gages for measurement of wall shear stress in Blastane experiments

NASA Technical Reports Server (NTRS)

Murthy, S. V.; Steinle, F. W.

1986-01-01

Buried Wire Gages operated from a Constant Temperature Anemometer System are among the special types of instrumentation to be used in the Boundary Layer Apparatus for Subsonic and Transonic flow Affected by Noise Environment (BLASTANE). These Gages are of a new type and need to be adapted for specific applications. Methods were developed to fabricate Gage inserts and mount those in the BLASTANE Instrumentation Plugs. A large number of Gages were prepared and operated from a Constant Temperature Anemometer System to derive some of the calibration constants for application to fluid-flow wall shear-stress measurements. The final stage of the calibration was defined, but could not be accomplished because of non-availability of a suitable flow simulating apparatus. This report provides a description of the Buried Wire Gage technique, an explanation of the method evolved for making proper Gages and the calibration constants, namely Temperature Coefficient of Resistance and Conduction Loss Factor.

Improved Regression Analysis of Temperature-Dependent Strain-Gage Balance Calibration Data

NASA Technical Reports Server (NTRS)

Ulbrich, N.

2015-01-01

An improved approach is discussed that may be used to directly include first and second order temperature effects in the load prediction algorithm of a wind tunnel strain-gage balance. The improved approach was designed for the Iterative Method that fits strain-gage outputs as a function of calibration loads and uses a load iteration scheme during the wind tunnel test to predict loads from measured gage outputs. The improved approach assumes that the strain-gage balance is at a constant uniform temperature when it is calibrated and used. First, the method introduces a new independent variable for the regression analysis of the balance calibration data. The new variable is designed as the difference between the uniform temperature of the balance and a global reference temperature. This reference temperature should be the primary calibration temperature of the balance so that, if needed, a tare load iteration can be performed. Then, two temperature{dependent terms are included in the regression models of the gage outputs. They are the temperature difference itself and the square of the temperature difference. Simulated temperature{dependent data obtained from Triumph Aerospace's 2013 calibration of NASA's ARC-30K five component semi{span balance is used to illustrate the application of the improved approach.

Strain Gage Signal Interpretation.

DTIC Science & Technology

1986-02-01

blades and vanes in many engines have been collected, played back and examined. The engine types encompass GE’s stable of turbine engines from the small...aeromechanical engineer . 1.3 SUMMARY OF RESULTS Strain gage signals from vibrating rotor blades and vanes were collected, examined, classified, and generalized...turboprops, to turbojets and to the large high bypass turbofan engines . Test conditions include all the phases that are investigated

Cost effectiveness of the U.S. Geological Survey's stream-gaging program in Illinois

USGS Publications Warehouse

Mades, D.M.; Oberg, K.A.

1984-01-01

Data uses and funding sources were identified for 138 continuous-record discharge-gaging stations currently (1983) operated as part of the stream-gaging program in Illinois. Streamflow data from five of those stations are used only for regional hydrology studies. Most streamflow data are used for defining regional hydrology, defining rainfall-runoff relations, flood forecasting, regulating navigation systems, and water-quality sampling. Based on the evaluations of data use and of alternative methods for determining streamflow in place of stream gaging, no stations in the 1983 stream-gaging program should be deactivated. The current budget (in 1983 dollars) for operating the 138-station program is $768,000 per year. The average standard error of instantaneous discharge for the current practice for visiting the gaging stations is 36.5 percent. Missing stage record accounts for one-third of the 36.5 percent average standard error. (USGS)

Evaluation of submarine strain-gage systems for monitoring coastal sediment migration

NASA Technical Reports Server (NTRS)

Shideler, G. L.; Mcgrath, D. G.

1973-01-01

Single and multiple strain-gage systems were respectively evaluated as in situ point and areal sensors for monitoring sand-height variations in coastal environments. Static loading tests indicate that gage response pressure is linear for sand heights up to 24 inches. Response pressures are a function of both sand height and aggregate density, with density being influenced by both sediment texture and degree of compaction. Poorer sediment sorting and greater compaction result in higher response pressures. Field tests in a beach foreshore environment indicate that the gage systems are effective qualitative instruments for monitoring long-period migration trends of beach sediments; whereas, short-period responses are not sufficiently reliable. The durability and compactness of the gage systems must be substantially increased for effective field operations. It is recommended that the systems' qualitative potentials be further developed, whereas their development as quantitative instruments be terminated. Further development should emphasize the construction of remote recording systems designed for semipermanent installation.

Levels at Streamflow Gaging Stations--A CD-ROM Based Training Class

USGS Publications Warehouse

Nolan, K. Michael; Jacobson, Nathan; Erickson, Robert; Landon, Stanley

2003-01-01

Streamgages record the elevation of the water surface above some reference surface, or datum. This datum is assumed to remain unchanged throughout the life of the gage. However, the elevation of gages and their supporting structures often change over time as a result of earthmovement, floods, ice, and debris. The surveying practice of leveling is used to establish datum for new gage structures and to check for vertical movement of those structures over time. Vertical changes in gage structures can affect stage-discharge relations and, thus, could result in incorrect discharge determinations. Datum checks are used to correct stage-discharge relations and allow the USGS to document gage datum throughout the life of a gage. This training presentation describes methods currently used by the U.S. Geological Survey to run levels at gaging stations. The presentation is narrated, but you control the pace of the presentation. If the computer you are using can view 'MPEG' videos you will be able to take advantage of videos found within the presentation. A test, found at the end of the presentation, can be taken to assess how well you understood the training material. The class is registered as class SW1307 with the National Training Center of the U.S. Geologcial Survey. The presentation was developed using Macromedia Director 8.5(1) and is contained in the file 'WRI-4002.exe', which should auto-launch after the CD-ROM is inserted in the PC. The program only runs on a windows-based personal computer (PC). A sound card and speakers are necessary to take advantage of the narration that accompanies the presentation. Text of narrations is provided, if you are unable to listen to the narrations. Instructions for installing and running the presentation are included in the file ' Intro.html'. The file 'Intro.html' is on the CD-ROM containing the presentation and is available from the presentation's help menu.

Gage measures total radiation, including vacuum UV, from ionized high-temperature gases

NASA Technical Reports Server (NTRS)

Wood, A. D.

1969-01-01

Transient-heat transfer gage measures the total radiation intensity from vacuum ultraviolet and ionized high temperature gases. The gage includes a sensitive piezoelectric crystal that is completely isolated from any ionized flow and vacuum ultraviolet irradiation.

Ceramic Strain Gages for Use at Temperatures up to 1500 Celsius

NASA Technical Reports Server (NTRS)

Gregory, Otto; Fralick, Gustave (Technical Monitor)

2003-01-01

Indium-tin-oxide (ITO) thin film strain gages were successfully demonstrated at temperatures beyond 1500 C. High temperature static strain tests revealed that the piezoresistive response and electrical stability of the ceramic sensors depended on the thickness of the ITO films comprising the active strain elements. When 2.5 microns-thick ITO films were employed as the active strain elements, the piezoresistive response became unstable at temperatures above 1225 C. In contrast to this, ceramic sensors prepared with 5 microns-thick ITO were stable beyond 1430 C and sensors prepared with 8 microns-thick ITO survived more than 20 hr of operation at 1481 C. Very thick (10 microns) ITo strain gages were extremely stable and responsive at 1528 C. ESCA depth profiles confirmed that an interfacial reaction between the ITO strain gage and alumina substrate was responsible for the high temperature electrical stability observed. Similar improvements in high temperature stability were achieved by doping the active ITO strain elements with aluminum. Several Sic-Sic CMC constant strain beams were instrumented with ITO strain gages and delivered to NASA for testing. Due to the extreme surface roughness of the CMC substrates, new lithography techniques and surface preparation methods were developed. These techniques relied heavily on a combination of Sic and A12O3 cement layers to provide the necessary surface finish for efficient pattern transfer. Micro-contact printing using soft lithography and PDMS stamps was also used to successfully transfer the thin film strain gage patterns to the resist coated CMC substrates. This latter approach has considerable potential for transferring the thin film strain gage patterns to the extremely rough surfaces associated with the CMC's.

Impact gages for detecting meteoroid and other orbital debris impacts on space vehicles.

NASA Technical Reports Server (NTRS)

Mastandrea, J. R.; Scherb, M. V.

1973-01-01

Impacts on space vehicles have been simulated using the McDonnell Douglas Aerophysics Laboratory (MDAL) Light-Gas Guns to launch particles at hypervelocity speeds into scaled space structures. Using impact gages and a triangulation technique, these impacts have been detected and accurately located. This paper describes in detail the various types of impact gages (piezoelectric PZT-5A, quartz, electret, and off-the-shelf plastics) used. This description includes gage design and experimental results for gages installed on single-walled scaled payload carriers, multiple-walled satellites and space stations, and single-walled full-scale Delta tank structures. A brief description of the triangulation technique, the impact simulation, and the data acquisition system are also included.

Problems Associated with Attaching Strain Gages to Titanium Alloy Ti-61-4V

NASA Technical Reports Server (NTRS)

Jenkins, J. M.; Lemcoe, M. M.

1977-01-01

Weldable strain gages have shown excellent high temperature characteristics for supersonic cruise aircraft application. The spotwelding attachment method, however, has resulted in serious reductions in the fatigue life of titanium alloy (Ti-6Al-4V) fatigue specimens. The reduction is so severe that the use of weldable strain gages on operational aircraft must be prohibited. The cause of the fatigue problem is thought to be a combination of the microstructure changes in the material caused by spotwelding and the presence of the flange of the stain gage. Brazing, plating, and plasma spraying were investigated as substitutes for spotwelding. The attachment of a flangeless gage by plasma spraying provided the most improvement in the fatigue life of the titanium.

An economical storage precipitation gage

Treesearch

Raymond D. Ratliff; Jack N. Reppert

1965-01-01

Describes how to make an economical "can-cone"-type storage precipitation gage, based on one originally designed by the California Department of Water Resources. The cone was modified by including an inner skirt to prevent loss of water between the cone and can. Periodic records of precipitation are obtained by stick measurement. Five complete cones cost $120...

High-Temperature Extensometry and PdCr Temperature-Compensated Wire Resistance Strain Gages Compared

NASA Technical Reports Server (NTRS)

1996-01-01

A detailed experimental evaluation is underway at the NASA Lewis Research Center to compare and contrast the performance of the PdCr/Pt dual-element temperature-compensated wire resistance strain gage with that of conventional high-temperature extensometry. The advanced PdCr gage, developed by researchers at Lewis, exhibits desirable properties and a relatively small and repeatable apparent strain to 800 C. This gage represents a significant advance in technology because existing commercial resistance strain gages are not reliable for quasi-static strain measurements above approximately 400 C. Various thermal and mechanical loading spectra are being applied by a high-temperature thermomechanical uniaxial testing system to evaluate the two strain-measurement systems. This is being done not only to compare and contrast the two strain sensors, but also to investigate the applicability of the PdCr strain gage to the coupon-level specimen testing environment typically employed when the high-temperature mechanical behavior of structural materials is characterized. Strain measurement capabilities to 800 C are being investigated with a nickel-base superalloy, Inconel 100 (IN 100), substrate material and application to TMC's is being examined with the model system, SCS-6/Ti-15-3. Furthermore, two gage application techniques are being investigated in the comparison study: namely, flame-sprayed and spot welding.

Analysis of streamflow-gaging network for monitoring stormwater in small streams in the Puget Sound Basin, Washington

USGS Publications Warehouse

Konrad, Christopher P.; Voss, Frank D.

2012-01-01

The streamflow-gaging network in the Puget Sound basin was analyzed for its capacity to monitor stormwater in small streams. The analysis consisted of an inventory of active and inactive gages and an evaluation of the coverage and resolution of the gaging network with an emphasis on lowland areas. The active gaging network covers much of the Puget Lowland largely by gages located at sites on larger streams and rivers. Assessments of stormwater impacts and management will likely require streamflow information with higher spatial resolution than provided by the current gaging network. Monitoring that emphasizes small streams in combination with approaches for estimating streamflow at ungaged sites provides an alternative to expanding the current gaging network that can improve the spatial resolution of streamflow information in the region. The highest priority gaps in the gaging network are low elevation basins close to the Puget Sound shoreline and sites that share less than 10 percent of the drainage area of an active gage. Although small, lowland sites with long records of streamflow are particularly valuable to maintain in the region, other criteria for prioritizing sites in the gaging network should be based on the specific questions that stormwater managers need to answer.

Evaluation of the streamflow-gaging network of Alaska in providing regional streamflow information

USGS Publications Warehouse

Brabets, Timothy P.

1996-01-01

In 1906, the U.S. Geological Survey (USGS) began operating a network of streamflow-gaging stations in Alaska. The primary purpose of the streamflow- gaging network has been to provide peak flow, average flow, and low-flow characteristics to a variety of users. In 1993, the USGS began a study to evaluate the current network of 78 stations. The objectives of this study were to determine the adequacy of the existing network in predicting selected regional flow characteristics and to determine if providing additional streamflow-gaging stations could improve the network's ability to predict these characteristics. Alaska was divided into six distinct hydrologic regions: Arctic, Northwest, Southcentral, Southeast, Southwest, and Yukon. For each region, historical and current streamflow data were compiled. In Arctic, Northwest, and Southwest Alaska, insufficient data were available to develop regional regression equations. In these areas, proposed locations of streamflow-gaging stations were selected by using clustering techniques to define similar areas within a region and by spatial visual analysis using the precipitation, physiographic, and hydrologic unit maps of Alaska. Sufficient data existed in Southcentral and Southeast Alaska to use generalized least squares (GLS) procedures to develop regional regression equations to estimate the 50-year peak flow, annual average flow, and a low-flow statistic. GLS procedures were also used for Yukon Alaska but the results should be used with caution because the data do not have an adequate spatial distribution. Network analysis procedures were used for the Southcentral, Southeast, and Yukon regions. Network analysis indicates the reduction in the sampling error of the regional regression equation that can be obtained given different scenarios. For Alaska, a 10-year planning period was used. One scenario showed the results of continuing the current network with no additional gaging stations and another scenario showed the results

Effectiveness of the New Hampshire stream-gaging network in providing regional streamflow information

USGS Publications Warehouse

Olson, Scott A.

2003-01-01

The stream-gaging network in New Hampshire was analyzed for its effectiveness in providing regional information on peak-flood flow, mean-flow, and low-flow frequency. The data available for analysis were from stream-gaging stations in New Hampshire and selected stations in adjacent States. The principles of generalized-least-squares regression analysis were applied to develop regional regression equations that relate streamflow-frequency characteristics to watershed characteristics. Regression equations were developed for (1) the instantaneous peak flow with a 100-year recurrence interval, (2) the mean-annual flow, and (3) the 7-day, 10-year low flow. Active and discontinued stream-gaging stations with 10 or more years of flow data were used to develop the regression equations. Each stream-gaging station in the network was evaluated and ranked on the basis of how much the data from that station contributed to the cost-weighted sampling-error component of the regression equation. The potential effect of data from proposed and new stream-gaging stations on the sampling error also was evaluated. The stream-gaging network was evaluated for conditions in water year 2000 and for estimated conditions under various network strategies if an additional 5 years and 20 years of streamflow data were collected. The effectiveness of the stream-gaging network in providing regional streamflow information could be improved for all three flow characteristics with the collection of additional flow data, both temporally and spatially. With additional years of data collection, the greatest reduction in the average sampling error of the regional regression equations was found for the peak- and low-flow characteristics. In general, additional data collection at stream-gaging stations with unregulated flow, relatively short-term record (less than 20 years), and drainage areas smaller than 45 square miles contributed the largest cost-weighted reduction to the average sampling error of the

The index gage method to develop a flow duration curve from short-term streamflow records

NASA Astrophysics Data System (ADS)

Zhang, Zhenxing

2017-10-01

The flow duration curve (FDC) is one of the most commonly used graphical tools in hydrology and provides a comprehensive graphical view of streamflow variability at a particular site. For a gaged site, an FDC can be easily estimated with frequency analysis. When no streamflow records are available, regional FDCs are used to synthesize FDCs. However, studies on how to develop FDCs for sites with short-term records have been very limited. Deriving representative FDC when there are short-term hydrologic records is important. For instance, 43% of the 394 streamflow gages in Illinois have records of 20 years or fewer, and these short-term gages are often distributed in headwaters and contain valuable hydrologic information. In this study, the index gage method is proposed to develop FDCs using short-term hydrologic records via an information transfer technique from a nearby hydrologically similar index gage. There are three steps: (1) select an index gage; (2) determine changes of FDC; and (3) develop representative FDCs. The approach is tested using records from 92 U.S. Geological Survey streamflow gages in Illinois. A jackknife experiment is conducted to assess the performance. Bootstrap resampling is used to simulate various periods of records, i.e., 1, 2, 5, 10, 15, and 20 years of records. The results demonstrated that the index gage method is capable of developing a representative FDC using short-term records. Generally, the approach performance is improved when more hydrologic records are available, but the improvement appears to level off when the short-term gage has 10 years or more records.

Matilda Joslyn Gage: A Nineteenth-Century Women's Rights Historian Looks at Witchcraft.

ERIC Educational Resources Information Center

Corey, Mary E.

2003-01-01

Explores the ideas of the nineteenth century female historian, Matilda Joslyn Gage, who authored the book, "Woman, Church, and State." Focuses on Gage's ideas about women's history, particularly related to the role of the church and women persecuted for witchcraft. (CMK)

Cost effectiveness of the US Geological Survey's stream-gaging programs in New Hampshire and Vermont

USGS Publications Warehouse

Smath, J.A.; Blackey, F.E.

1986-01-01

Data uses and funding sources were identified for the 73 continuous stream gages currently (1984) being operated. Eight stream gages were identified as having insufficient reason to continue their operation. Parts of New Hampshire and Vermont were identified as needing additional hydrologic data. New gages should be established in these regions as funds become available. Alternative methods for providing hydrologic data at the stream gaging stations currently being operated were found to lack the accuracy that is required for their intended use. The current policy for operation of the stream gages requires a net budget of $297,000/yr. The average standard error of estimation of the streamflow records is 17.9%. This overall level of accuracy could be maintained with a budget of $285,000 if resources were redistributed among gages. Cost-effective analysis indicates that with the present budget, the average standard error could be reduced to 16.6%. A minimum budget of $278,000 is required to operate the present stream gaging program. Below this level, the gages and recorders would not receive the proper service and maintenance. At the minimum budget, the average standard error would be 20.4%. The loss of correlative data is a significant component of the error in streamflow records, especially at lower budgetary levels. (Author 's abstract)

In-Flight Capability for Evaluating Skin-Friction Gages and Other Near-Wall Flow Sensors

NASA Technical Reports Server (NTRS)

Bui, Trong T.; Pipitone, Brett J.; Krake, Keith L.; Richwine, Dave (Technical Monitor)

2003-01-01

An 8-in.-square boundary-layer sensor panel has been developed for in-flight evaluation of skin-friction gages and other near-wall flow sensors on the NASA Dryden Flight Research Center F-15B/Flight Test Fixture (FTF). Instrumentation on the sensor panel includes a boundary-layer rake, temperature sensors, static pressure taps, and a Preston tube. Space is also available for skin-friction gages or other near-wall flow sensors. Pretest analysis of previous F-15B/FTF flight data has identified flight conditions suitable for evaluating skin-friction gages. At subsonic Mach numbers, the boundary layer over the sensor panel closely approximates the two-dimensional (2D), law-of-the-wall turbulent boundary layer, and skin-friction estimates from the Preston tube and the rake (using the Clauser plot method) can be used to evaluate skin-friction gages. At supersonic Mach numbers, the boundary layer over the sensor panel becomes complex, and other means of measuring skin friction are needed to evaluate the accuracy of new skin-friction gages. Results from the flight test of a new rubber-damped skin-friction gage confirm that at subsonic Mach numbers, nearly 2D, law-of-the-wall turbulent boundary layers exist over the sensor panel. Sensor panel data also show that this new skin-friction gage prototype does not work in flight.

Analysis of strain gage reliability in F-100 jet engine testing at NASA Lewis Research Center

NASA Technical Reports Server (NTRS)

Holanda, R.

1983-01-01

A reliability analysis was performed on 64 strain gage systems mounted on the 3 rotor stages of the fan of a YF-100 engine. The strain gages were used in a 65 hour fan flutter research program which included about 5 hours of blade flutter. The analysis was part of a reliability improvement program. Eighty-four percent of the strain gages survived the test and performed satisfactorily. A post test analysis determined most failure causes. Five failures were caused by open circuits, three failed gages showed elevated circuit resistance, and one gage circuit was grounded. One failure was undetermined.

Cost-effectiveness of the stream-gaging program in New Jersey

USGS Publications Warehouse

Schopp, R.D.; Ulery, R.L.

1984-01-01

The results of a study of the cost-effectiveness of the stream-gaging program in New Jersey are documented. This study is part of a 5-year nationwide analysis undertaken by the U.S. Geological Survey to define and document the most cost-effective means of furnishing streamflow information. This report identifies the principal uses of the data and relates those uses to funding sources, applies, at selected stations, alternative less costly methods (that is flow routing, regression analysis) for furnishing the data, and defines a strategy for operating the program which minimizes uncertainty in the streamflow data for specific operating budgets. Uncertainty in streamflow data is primarily a function of the percentage of missing record and the frequency of discharge measurements. In this report, 101 continuous stream gages and 73 crest-stage or stage-only gages are analyzed. A minimum budget of $548,000 is required to operate the present stream-gaging program in New Jersey with an average standard error of 27.6 percent. The maximum budget analyzed was $650,000, which resulted in an average standard error of 17.8 percent. The 1983 budget of $569,000 resulted in a standard error of 24.9 percent under present operating policy. (USGS)

Cost-effectiveness of the stream-gaging program in North Carolina

USGS Publications Warehouse

Mason, R.R.; Jackson, N.M.

1985-01-01

This report documents the results of a study of the cost-effectiveness of the stream-gaging program in North Carolina. Data uses and funding sources are identified for the 146 gaging stations currently operated in North Carolina with a budget of $777,600 (1984). As a result of the study, eleven stations are nominated for discontinuance and five for conversion from recording to partial-record status. Large parts of North Carolina 's Coastal Plain are identified as having sparse streamflow data. This sparsity should be remedied as funds become available. Efforts should also be directed toward defining the efforts of drainage improvements on local hydrology and streamflow characteristics. The average standard error of streamflow records in North Carolina is 18.6 percent. This level of accuracy could be improved without increasing cost by increasing the frequency of field visits and streamflow measurements at stations with high standard errors and reducing the frequency at stations with low standard errors. A minimum budget of $762,000 is required to operate the 146-gage program. A budget less than this does not permit proper service and maintenance of the gages and recorders. At the minimum budget, and with the optimum allocation of field visits, the average standard error is 17.6 percent.

Strain Gage Measurements of Aft Nacelle Shock Absorbers.

DTIC Science & Technology

ENGINE NACELLES, SHOCK ABSORBERS ), (* SHOCK ABSORBERS , STRESSES), SURFACE TO SURFACE MISSILES, LAUNCHING, STRAIN GAGES, COMPRESSIVE PROPERTIES, CALIBRATION, STRAIN(MECHANICS), FAILURE, GROUND SUPPORT EQUIPMENT.

Detection of Bi-Directionality in Strain-Gage Balance Calibration Data

NASA Technical Reports Server (NTRS)

Ulbrich, Norbert

2012-01-01

An indicator variable was developed for both visualization and detection of bi-directionality in wind tunnel strain-gage balance calibration data. First, the calculation of the indicator variable is explained in detail. Then, a criterion is discussed that may be used to decide which gage outputs of a balance have bi- directional behavior. The result of this analysis could be used, for example, to justify the selection of certain absolute value or other even function terms in the regression model of gage outputs whenever the Iterative Method is chosen for the balance calibration data analysis. Calibration data of NASA s MK40 Task balance is analyzed to illustrate both the calculation of the indicator variable and the application of the proposed criterion. Finally, bi directionality characteristics of typical multi piece, hybrid, single piece, and semispan balances are determined and discussed.

Rainfall, streamflow, and peak stage data collected at the Murfreesboro, Tennessee, gaging network, March 1989 through July 1992

USGS Publications Warehouse

Outlaw, G.S.; Butner, D.E.; Kemp, R.L.; Oaks, A.T.; Adams, G.S.

1992-01-01

Rainfall, stage, and streamflow data in the Murfreesboro area, Middle Tennessee, were collected from March 1989 through July 1992 from a network of 68 gaging stations. The network consists of 10 tipping-bucket rain gages, 2 continuous-record streamflow gages, 4 partial-record flood hydrograph gages, and 72 crest-stage gages. Data collected by the gages includes 5minute time-step rainfall hyetographs, 15-minute time-step flood hydrographs, and peak-stage elevations. Data are stored in a computer data base and are available for many computer modeling and engineering applications.

An Elastic Tube Gage for Measuring Static and Dynamic Pressures

DTIC Science & Technology

1948-05-01

AUXILIARY HEAT RADIATORS - .-. • • /•’ • • • AUXILIARY ELECTRICAL AND RECORDING EQUIPMENT . . . . .. RESULTS OF FIELD TESTS...time and resources and to permit a greater flexibility in use where measurements are required of a large number or variety of quantities.. Since...gages are first prercürved to fit the tube., and then cemented with Duco. After air-drying for H8 hours, the gages, are heated with infrared lamps to

Calibration and evaluation tests of strain gages for use on structure exposed to cryogenic and reentry temperatures

NASA Astrophysics Data System (ADS)

Mueller, Richard N.; Howard, J. Lawrence; Sikorra, Charles F.; Swegle, Allan R.

Commercial strain gages were evaluated for proposed strain measurement on a Rene 41 honeycomb test panel to be subjected to temperatures from -423 F to +1600 F. Foil strain gages of three different temperature compensations, a weldable strain gage, and a capacitive strain gage, were tested to determine characteristics of apparent strain, strain sensitivity, and temperature operational limits under stabilized temperature and several heating and cooling temperature rates. Test results show that strain measurement over the total temperature range can be made using a combination of gages.

Latin-square three-dimensional gage master

DOEpatents

Jones, L.

1981-05-12

A gage master for coordinate measuring machines has an nxn array of objects distributed in the Z coordinate utilizing the concept of a Latin square experimental design. Using analysis of variance techniques, the invention may be used to identify sources of error in machine geometry and quantify machine accuracy.

Latin square three dimensional gage master

DOEpatents

Jones, Lynn L.

1982-01-01

A gage master for coordinate measuring machines has an nxn array of objects distributed in the Z coordinate utilizing the concept of a Latin square experimental design. Using analysis of variance techniques, the invention may be used to identify sources of error in machine geometry and quantify machine accuracy.

High temperature static strain gage development

NASA Technical Reports Server (NTRS)

Hulse, C. O.; Bailey, R. S.; Grant, H. P.; Anderson, W. L.; Przybyszewski, J. S.

1991-01-01

Final results are presented from a program to develop a thin film static strain gage for use on the blades and vanes of running, test stand gas turbine engines with goals of an 3 x 3 mm gage area and total errors of less than 10 pct. of + or - 2,000 microstrain after 50 hrs at 1250 K. Pd containing 13 Wt. pct. Cr was previously identified as a new strain sensor alloy that appeared to be potentially usable to 1250 K. Subsequently, it was discovered, in contrast with its behavior in bulk, that Pd-13Cr suffered from oxidation attack when prepared as a 4.5 micron thick thin film. Continuing problems with electrical leakage to the substrate and the inability of sputtered alumina overcoats to prevent oxidation led to the discovery that sputtered alumina contains appreciable amounts of entrapped argon. After the argon has been exsolved by heating to elevated temperatures, the alumina films undergo a linear shrinkage of about 2 pct. resulting in formation of cracks. These problems can be largely overcome by sputtering the alumina with the substrate heated to 870 K. With 2 micron thick hot sputtered alumina insulation and overcoat films, total 50 hr drifts of about 100 microstrain (2 tests) and about 500 microstrain (1 test) were observed at 1000 and 1100 K, respectively. Results of tests on complete strain gage systems on constant moment bend bars with Pd temperature compensation grids revealed that oxidation of the Pd grid was a major problem even when the grid was overcoated with a hot or cold sputtered alumina overcoat.

Evaluation of Troxler model 3411 nuclear gage.

DOT National Transportation Integrated Search

1978-01-01

The performance of the Troxler Electronics Laboratory Model 3411 nuclear gage was evaluated through laboratory tests on the Department's density and moisture standards and field tests on various soils, base courses, and bituminous concrete overlays t...

Cost-effectiveness of the Federal stream-gaging program in Virginia

USGS Publications Warehouse

Carpenter, D.H.

1985-01-01

Data uses and funding sources were identified for the 77 continuous stream gages currently being operated in Virginia by the U.S. Geological Survey with a budget of $446,000. Two stream gages were identified as not being used sufficiently to warrant continuing their operation. Operation of these stations should be considered for discontinuation. Data collected at two other stations were identified as having uses primarily related to short-term studies; these stations should also be considered for discontinuation at the end of the data collection phases of the studies. The remaining 73 stations should be kept in the program for the foreseeable future. The current policy for operation of the 77-station program requires a budget of $446,000/yr. The average standard error of estimation of streamflow records is 10.1%. It was shown that this overall level of accuracy at the 77 sites could be maintained with a budget of $430,500 if resources were redistributed among the gages. A minimum budget of $428,500 is required to operate the 77-gage program; a smaller budget would not permit proper service and maintenance of the gages and recorders. At the minimum budget, with optimized operation, the average standard error would be 10.4%. The maximum budget analyzed was $650,000, which resulted in an average standard error of 5.5%. The study indicates that a major component of error is caused by lost or missing data. If perfect equipment were available, the standard error for the current program and budget could be reduced to 7.6%. This also can be interpreted to mean that the streamflow data have a standard error of this magnitude during times when the equipment is operating properly. (Author 's abstract)

Cost effective stream-gaging strategies for the Lower Colorado River basin; the Blythe field office operations

USGS Publications Warehouse

Moss, Marshall E.; Gilroy, Edward J.

1980-01-01

This report describes the theoretical developments and illustrates the applications of techniques that recently have been assembled to analyze the cost-effectiveness of federally funded stream-gaging activities in support of the Colorado River compact and subsequent adjudications. The cost effectiveness of 19 stream gages in terms of minimizing the sum of the variances of the errors of estimation of annual mean discharge is explored by means of a sequential-search optimization scheme. The search is conducted over a set of decision variables that describes the number of times that each gaging route is traveled in a year. A gage route is defined as the most expeditious circuit that is made from a field office to visit one or more stream gages and return to the office. The error variance is defined as a function of the frequency of visits to a gage by using optimal estimation theory. Currently a minimum of 12 visits per year is made to any gage. By changing to a six-visit minimum, the same total error variance can be attained for the 19 stations with a budget of 10% less than the current one. Other strategies are also explored. (USGS)

HOLEGAGE 1.0 - STRAIN GAGE HOLE DRILLING ANALYSIS PROGRAM

NASA Technical Reports Server (NTRS)

Hampton, R. W.

1994-01-01

There is no simple and perfect way to measure residual stresses in metal parts that have been welded or deformed to make complex structures such as pressure vessels and aircraft, yet these locked-in stresses can contribute to structural failure by fatigue and fracture. However, one proven and tested technique for determining the internal stress of a metal part is to drill a test hole while measuring the relieved strains around the hole, such as the hole-drilling strain gage method described in ASTM E 837. The program HOLEGAGE processes strain gage data and provides additional calculations of internal stress variations that are not obtained with standard E 837 analysis methods. The typical application of the technique uses a three gage rosette with a special hole-drilling fixture for drilling a hole through the center of the rosette to produce a hole with very small gage pattern eccentricity error. Another device is used to control the drilling and halt the drill at controlled depth steps. At each step, strains from all three strain gages are recorded. The influence coefficients used by HOLEGAGE to compute stresses from relieved hole strains were developed by published finite element method studies of thick plates for specific hole sizes and depths. The program uses a parabolic fit and an interpolating scheme to project the coefficients to other hole sizes and depths. Additionally, published experimental data are used to extend the coefficients to relatively thin plates. These influence coefficients are used to compute the stresses in the original part from the strain data. HOLEGAGE will compute interior planar stresses using strain data from each drilled hole depth layer. Planar stresses may be computed in three ways including: a least squares fit for a linear variation with depth, an integral method to give incremental stress data for each layer, or by a linear fit to the integral data (with some surface data points omitted) to predict surface stresses before

Comparative Protein Profiling of Intraphagosomal Expressed Proteins of Mycobacterium bovis BCG.

PubMed

Singhal, Neelja; Kumar, Manish; Sharma, Divakar; Bisht, Deepa

2016-01-01

BCG, the only available vaccine against tuberculosis affords a variable protection which wanes with time. In this study we have analyzed and compared the proteins which are expressed differentially during broth-culture and intraphagosomal growth of M.bovis BCG. Eight proteins which showed increased expression during the intraphagosomal growth were identified by MALDI-TOF/MS. These were - a precursor of alanine and proline-rich secreted protein apa, isoforms of malate dehydrogenase, large subunit alpha (Alpha-ETF) of electron transfer flavoprotein, immunogenic protein MPB64 precursor, UPF0036 protein, and two proteins with unknown function. Based on these findings we speculate that higher expression of these proteins has a probable role in intracellular survival, adaptation and/or immunoprotective effect of BCG. Further, these proteins might also be used as gene expression markers for endosome trafficking events of BCG.

Detection and Use of Load and Gage Output Repeats of Wind Tunnel Strain-Gage Balance Data

NASA Technical Reports Server (NTRS)

Ulbrich, N.

2017-01-01

Criteria are discussed that may be used for the detection of load and gage output repeats of wind tunnel strain-gage balance data. First, empirical thresholds are introduced that help determine if the loads or electrical outputs of a pair of balance calibration or check load data points match. A threshold of 0.01 percent of the load capacity is suggested for the identification of matching loads. Similarly, a threshold of 0.1 microV/V is recommended for the identification of matching electrical outputs. Two examples for the use of load and output repeats are discussed to illustrate benefits of the implementation of a repeat point detection algorithm in a balance data analysis software package. The first example uses the suggested load threshold to identify repeat data points that may be used to compute pure errors of the balance loads. This type of analysis may reveal hidden data quality issues that could potentially be avoided by making calibration process improvements. The second example uses the electrical output threshold for the identification of balance fouling. Data from the calibration of a six-component force balance is used to illustrate the calculation of the pure error of the balance loads.

Analysis of the streamflow-gaging station network in Ohio for effectiveness in providing regional streamflow information

USGS Publications Warehouse

Straub, D.E.

1998-01-01

The streamflow-gaging station network in Ohio was evaluated for its effectiveness in providing regional streamflow information. The analysis involved application of the principles of generalized least squares regression between streamflow and climatic and basin characteristics. Regression equations were developed for three flow characteristics: (1) the instantaneous peak flow with a 100-year recurrence interval (P100), (2) the mean annual flow (Qa), and (3) the 7-day, 10-year low flow (7Q10). All active and discontinued gaging stations with 5 or more years of unregulated-streamflow data with respect to each flow characteristic were used to develop the regression equations. The gaging-station network was evaluated for the current (1996) condition of the network and estimated conditions of various network strategies if an additional 5 and 20 years of streamflow data were collected. Any active or discontinued gaging station with (1) less than 5 years of unregulated-streamflow record, (2) previously defined basin and climatic characteristics, and (3) the potential for collection of more unregulated-streamflow record were included in the network strategies involving the additional 5 and 20 years of data. The network analysis involved use of the regression equations, in combination with location, period of record, and cost of operation, to determine the contribution of the data for each gaging station to regional streamflow information. The contribution of each gaging station was based on a cost-weighted reduction of the mean square error (average sampling-error variance) associated with each regional estimating equation. All gaging stations included in the network analysis were then ranked according to their contribution to the regional information for each flow characteristic. The predictive ability of the regression equations developed from the gaging station network could be improved for all three flow characteristics with the collection of additional streamflow data

Identifying dissolved organic carbon sources at a gaged headwater catchment using FDOM sensors

NASA Astrophysics Data System (ADS)

Malzone, J. M.; Shanley, J. B.

2014-12-01

The United States Geological Survey's (USGS) W-9 gage at the headwaters of Sleepers River, Vermont has been monitored for dissolved organic carbon (DOC) concentration for more than 20 years. However, the sources of this DOC during base flow and hydrologic events remain unclear. The major objectives of this research were to identify sources of DOC during storm events and to explain the observed DOC-streamflow counterclockwise hysteresis during hydrologic events. Two main hypotheses to explain hysteresis during hydrologic events were tested: (1) distant headwater wetlands are the major DOC source, which lags behind peak flow due to travel time; and (2) the entire watershed contributes to the DOC at the gage, but the response of DOC lags behind the period when groundwater contributes most to streamflow. Sources of DOC were tracked using fluorescent dissolved organic matter (FDOM) sensors in surface water and groundwater wells. Wells were installed at four depths, 0.3, 0.6, 0.9, and 1.2 m, at four sites: a peaty low-gradient riparian area near the headwaters; a mid-hillslope area on a long hillslope mid-watershed; a near-stream area on a long hillslope mid-watershed; and a low-gradient tributary confluence area just above the gage. During storm events, FDOM and hydraulic head were measured at the nested groundwater wells. Samples for DOC analysis were also taken to determine the relationship between FDOM and DOC. Results suggest that both distant sources and the greater watershed played a role in the transport of DOC to the W-9 gage. Distant peaty sources dominated during large storms and contributed the highest surface water FDOM measurements. The peak FDOM at the gage was therefore best described as a result of transport. However, export from these distant sources terminated rapidly and did not explain continued elevated FDOM at the gage. Groundwater across the watershed exhibited hysteresis analogous to that in the stream itself, with FDOM peaking as head receded

Characteristics of Extrinsic Fabry-Perot Interferometric (EFPI) Fiber-Optic Strain Gages

NASA Technical Reports Server (NTRS)

Hare, David A.; Moore, Thomas C., Sr.

2000-01-01

The focus of this paper is a comparison of the strain-measuring characteristics of one type of commercially available fiber-optic strain sensor with the performance of conventional resistance strain gages. Fabry-Perot type fiber-optic strain sensors were selected for this testing program. Comparative testing is emphasized and includes load testing at room temperature with apparent strain characterization cryogenically and at elevated temperatures. The absolute accuracy of either of these types of strain gages is not addressed.

Evaluation of a nuclear gage for controlling the consolidation of fresh concrete : final report.

DOT National Transportation Integrated Search

1981-01-01

Evaluated was an approach to controlling the consolidation of in-place portland cement concrete on the basis of densities obtained with a Troxler 3411 nuclear gage. The gage was used in the backscatter mode on low-slump concrete bridge deck overlays ...

Mapping connectivity damage in the case of Phineas Gage.

PubMed

Van Horn, John Darrell; Irimia, Andrei; Torgerson, Carinna M; Chambers, Micah C; Kikinis, Ron; Toga, Arthur W

2012-01-01

White matter (WM) mapping of the human brain using neuroimaging techniques has gained considerable interest in the neuroscience community. Using diffusion weighted (DWI) and magnetic resonance imaging (MRI), WM fiber pathways between brain regions may be systematically assessed to make inferences concerning their role in normal brain function, influence on behavior, as well as concerning the consequences of network-level brain damage. In this paper, we investigate the detailed connectomics in a noted example of severe traumatic brain injury (TBI) which has proved important to and controversial in the history of neuroscience. We model the WM damage in the notable case of Phineas P. Gage, in whom a "tamping iron" was accidentally shot through his skull and brain, resulting in profound behavioral changes. The specific effects of this injury on Mr. Gage's WM connectivity have not previously been considered in detail. Using computed tomography (CT) image data of the Gage skull in conjunction with modern anatomical MRI and diffusion imaging data obtained in contemporary right handed male subjects (aged 25-36), we computationally simulate the passage of the iron through the skull on the basis of reported and observed skull fiducial landmarks and assess the extent of cortical gray matter (GM) and WM damage. Specifically, we find that while considerable damage was, indeed, localized to the left frontal cortex, the impact on measures of network connectedness between directly affected and other brain areas was profound, widespread, and a probable contributor to both the reported acute as well as long-term behavioral changes. Yet, while significantly affecting several likely network hubs, damage to Mr. Gage's WM network may not have been more severe than expected from that of a similarly sized "average" brain lesion. These results provide new insight into the remarkable brain injury experienced by this noteworthy patient.

Calibration of strain-gage installations in aircraft structures for the measurement of flight loads

NASA Technical Reports Server (NTRS)

Skopinski, T H; Aiken, William S , Jr; Huston, Wilber B

1954-01-01

A general method has been developed for calibrating strain-gage installations in aircraft structures, which permits the measurement in flight of the shear or lift, the bending moment, and the torque or pitching moment on the principal lifting or control surfaces. Although the stress in structural members may not be a simple function of the three loads of interest, a straightforward procedure is given for numerically combining the outputs of several bridges in such a way that the loads may be obtained. Extensions of the basic procedure by means of electrical combination of the strain-gage bridges are described which permit compromises between strain-gage installation time, availability of recording instruments, and data reduction time. The basic principles of strain-gage calibration procedures are illustrated by reference to the data for two aircraft structures of typical construction, one a straight and the other a swept horizontal stabilizer.

Cost-effectiveness of the streamflow-gaging program in Wyoming

USGS Publications Warehouse

Druse, S.A.; Wahl, K.L.

1988-01-01

This report documents the results of a cost-effectiveness study of the streamflow-gaging program in Wyoming. Regression analysis or hydrologic flow-routing techniques were considered for 24 combinations of stations from a 139-station network operated in 1984 to investigate suitability of techniques for simulating streamflow records. Only one station was determined to have sufficient accuracy in the regression analysis to consider discontinuance of the gage. The evaluation of the gaging-station network, which included the use of associated uncertainty in streamflow records, is limited to the nonwinter operation of the 47 stations operated by the Riverton Field Office of the U.S. Geological Survey. The current (1987) travel routes and measurement frequencies require a budget of $264,000 and result in an average standard error in streamflow records of 13.2%. Changes in routes and station visits using the same budget, could optimally reduce the standard error by 1.6%. Budgets evaluated ranged from $235,000 to $400,000. A $235,000 budget increased the optimal average standard error/station from 11.6 to 15.5%, and a $400,000 budget could reduce it to 6.6%. For all budgets considered, lost record accounts for about 40% of the average standard error. (USGS)

Cost-effectiveness of the stream-gaging program in Maine; a prototype for nationwide implementation

USGS Publications Warehouse

Fontaine, Richard A.; Moss, M.E.; Smath, J.A.; Thomas, W.O.

1984-01-01

This report documents the results of a cost-effectiveness study of the stream-gaging program in Maine. Data uses and funding sources were identified for the 51 continuous stream gages currently being operated in Maine with a budget of $211,000. Three stream gages were identified as producing data no longer sufficiently needed to warrant continuing their operation. Operation of these stations should be discontinued. Data collected at three other stations were identified as having uses specific only to short-term studies; it is recommended that these stations be discontinued at the end of the data-collection phases of the studies. The remaining 45 stations should be maintained in the program for the foreseeable future. The current policy for operation of the 45-station program would require a budget of $180,300 per year. The average standard error of estimation of streamflow records is 17.7 percent. It was shown that this overall level of accuracy at the 45 sites could be maintained with a budget of approximately $170,000 if resources were redistributed among the gages. A minimum budget of $155,000 is required to operate the 45-gage program; a smaller budget would not permit proper service and maintenance of the gages and recorders. At the minimum budget, the average standard error is 25.1 percent. The maximum budget analyzed was $350,000, which resulted in an average standard error of 8.7 percent. Large parts of Maine's interior were identified as having sparse streamflow data. It was determined that this sparsity be remedied as funds become available.

Improving membrane protein expression and function using genomic edits

DOE PAGES

Jensen, Heather M.; Eng, Thomas; Chubukov, Victor; ...

2017-10-12

Expression of membrane proteins often leads to growth inhibition and perturbs central metabolism and this burden varies with the protein being overexpressed. There are also known strain backgrounds that allow greater expression of membrane proteins but that differ in efficacy across proteins. Here, we hypothesized that for any membrane protein, it may be possible to identify a modified strain background where its expression can be accommodated with less burden. To directly test this hypothesis, we used a bar-coded transposon insertion library in tandem with cell sorting to assess genome-wide impact of gene deletions on membrane protein expression. The expression ofmore » five membrane proteins (CyoB, CydB, MdlB, YidC, and LepI) and one soluble protein (GST), each fused to GFP, was examined. We identified Escherichia coli mutants that demonstrated increased membrane protein expression relative to that in wild type. For two of the proteins (CyoB and CydB), we conducted functional assays to confirm that the increase in protein expression also led to phenotypic improvement in function. This study represents a systematic approach to broadly identify genetic loci that can be used to improve membrane protein expression, and our method can be used to improve expression of any protein that poses a cellular burden.« less

Improving membrane protein expression and function using genomic edits

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jensen, Heather M.; Eng, Thomas; Chubukov, Victor

Expression of membrane proteins often leads to growth inhibition and perturbs central metabolism and this burden varies with the protein being overexpressed. There are also known strain backgrounds that allow greater expression of membrane proteins but that differ in efficacy across proteins. Here, we hypothesized that for any membrane protein, it may be possible to identify a modified strain background where its expression can be accommodated with less burden. To directly test this hypothesis, we used a bar-coded transposon insertion library in tandem with cell sorting to assess genome-wide impact of gene deletions on membrane protein expression. The expression ofmore » five membrane proteins (CyoB, CydB, MdlB, YidC, and LepI) and one soluble protein (GST), each fused to GFP, was examined. We identified Escherichia coli mutants that demonstrated increased membrane protein expression relative to that in wild type. For two of the proteins (CyoB and CydB), we conducted functional assays to confirm that the increase in protein expression also led to phenotypic improvement in function. This study represents a systematic approach to broadly identify genetic loci that can be used to improve membrane protein expression, and our method can be used to improve expression of any protein that poses a cellular burden.« less

Refinement of moisture calibration curves for nuclear gage.

DOT National Transportation Integrated Search

1973-01-01

Over the last three years the Virginia Highway Research Council has directed a research effort toward improving the method of determining the moisture content of soils with a nuclear gage. The first task in this research was the determination of the ...

Silicon strain gages bonded on stainless steel using glass frit for strain sensor applications

NASA Astrophysics Data System (ADS)

Zhang, Zongyang; Cheng, Xingguo; Leng, Yi; Cao, Gang; Liu, Sheng

2014-05-01

In this paper, a steel pressure sensor using strain gages bonded on a 17-4 PH stainless steel (SS) diaphragm based on glass frit technology is proposed. The strain gages with uniform resistance are obtained by growing an epi-silicon layer on a single crystal silicon wafer using epitaxial deposition technique. The inorganic glass frits are used as the bonding material between the strain gages and the 17-4 PH SS diaphragm. Our results show that the output performances of sensors at a high temperature of 125 °C are almost equal those at room temperature, which indicates that the glass frit bonding is a good method and may lead to a significant advance in the high temperature applicability of silicon strain gage sensors. Finally, the microstructure of the cured organic adhesive and the fired glass frit are compared. It may be concluded that the defects of the cured organic adhesive deteriorate the hysteresis and repeatability errors of the sensors.

A New Load Residual Threshold Definition for the Evaluation of Wind Tunnel Strain-Gage Balance Data

NASA Technical Reports Server (NTRS)

Ulbrich, N.; Volden, T.

2016-01-01

A new definition of a threshold for the detection of load residual outliers of wind tunnel strain-gage balance data was developed. The new threshold is defined as the product between the inverse of the absolute value of the primary gage sensitivity and an empirical limit of the electrical outputs of a strain{gage. The empirical limit of the outputs is either 2.5 microV/V for balance calibration or check load residuals. A reduced limit of 0.5 microV/V is recommended for the evaluation of differences between repeat load points because, by design, the calculation of these differences removes errors in the residuals that are associated with the regression analysis of the data itself. The definition of the new threshold and different methods for the determination of the primary gage sensitivity are discussed. In addition, calibration data of a six-component force balance and a five-component semi-span balance are used to illustrate the application of the proposed new threshold definition to different types of strain{gage balances. During the discussion of the force balance example it is also explained how the estimated maximum expected output of a balance gage can be used to better understand results of the application of the new threshold definition.

Fluid quantity gaging

NASA Technical Reports Server (NTRS)

Mord, Allan J.; Snyder, Howard A.; Kilpatrick, Kathleen A.; Hermanson, Lynn A.; Hopkins, Richard A.; Vangundy, Donald A.

1988-01-01

A system for measuring the mass of liquid in a tank on orbit with 1 percent accuracy was developed and demonstrated. An extensive tradeoff identified adiabatic compression as the only gaging technique that is independent of gravity or its orientation, and of the size and distribution of bubbles in the tank. This technique is applicable to all Earth-storable and cryogenic liquids of interest for Space Station use, except superfluid helium, and can be applied to tanks of any size, shape, or internal structure. Accuracy of 0.2 percent was demonstrated in the laboratory, and a detailed analytical model was developed and verified by testing. A flight system architecture is presented that allows meeting the needs of a broad range of space fluid systems without custom development for each user.

Low TCR nanocomposite strain gages

NASA Technical Reports Server (NTRS)

Gregory, Otto J. (Inventor); Chen, Ximing (Inventor)

2012-01-01

A high temperature thin film strain gage sensor capable of functioning at temperatures above 1400.degree. C. The sensor contains a substrate, a nanocomposite film comprised of an indium tin oxide alloy, zinc oxide doped with alumina or other oxide semiconductor and a refractory metal selected from the group consisting of Pt, Pd, Rh, Ni, W, Ir, NiCrAlY and NiCoCrAlY deposited onto the substrate to form an active strain element. The strain element being responsive to an applied force.

Wind Tunnel Strain-Gage Balance Calibration Data Analysis Using a Weighted Least Squares Approach

NASA Technical Reports Server (NTRS)

Ulbrich, N.; Volden, T.

2017-01-01

A new approach is presented that uses a weighted least squares fit to analyze wind tunnel strain-gage balance calibration data. The weighted least squares fit is specifically designed to increase the influence of single-component loadings during the regression analysis. The weighted least squares fit also reduces the impact of calibration load schedule asymmetries on the predicted primary sensitivities of the balance gages. A weighting factor between zero and one is assigned to each calibration data point that depends on a simple count of its intentionally loaded load components or gages. The greater the number of a data point's intentionally loaded load components or gages is, the smaller its weighting factor becomes. The proposed approach is applicable to both the Iterative and Non-Iterative Methods that are used for the analysis of strain-gage balance calibration data in the aerospace testing community. The Iterative Method uses a reasonable estimate of the tare corrected load set as input for the determination of the weighting factors. The Non-Iterative Method, on the other hand, uses gage output differences relative to the natural zeros as input for the determination of the weighting factors. Machine calibration data of a six-component force balance is used to illustrate benefits of the proposed weighted least squares fit. In addition, a detailed derivation of the PRESS residuals associated with a weighted least squares fit is given in the appendices of the paper as this information could not be found in the literature. These PRESS residuals may be needed to evaluate the predictive capabilities of the final regression models that result from a weighted least squares fit of the balance calibration data.

Clip gage attachment for frictionless measurement of displacement during high-temperature mechanical testing

DOEpatents

Alexander, David J.

1994-01-01

An attachment for placement between a test specimen and a remote clip gage extensometer providing improved fracture toughness tests of materials at elevated temperature. Using a cylindrical tube and axial rod in new relationship, the device transfers the displacement signal of the fracture toughness test specimen directly to a clip gage extensometer located outside the high temperature furnace. Virtually frictionless operation is assured by having the test specimen center one end of the rod in one end of the tube, while the clip gage extensometer arms center the other end of the rod in the other end of the tube. By providing positive control over both ends of both rod and tube, the attachment may be operated in orientations other than vertical.

Clip gage attachment for frictionless measurement of displacement during high-temperature mechanical testing

DOEpatents

Alexander, D.J.

1994-01-04

An attachment for placement between a test specimen and a remote clip gage extensometer providing improved fracture toughness tests of materials at elevated temperature is described. Using a cylindrical tube and axial rod in new relationship, the device transfers the displacement signal of the fracture toughness test specimen directly to a clip gage extensometer located outside the high temperature furnace. Virtually frictionless operation is assured by having the test specimen center one end of the rod in one end of the tube, while the clip gage extensometer arms center the other end of the rod in the other end of the tube. By providing positive control over both ends of both rod and tube, the attachment may be operated in orientations other than vertical. 1 figure.

Evaluation of the cost effectiveness of the 1983 stream-gaging program in Kansas

USGS Publications Warehouse

Medina, K.D.; Geiger, C.O.

1984-01-01

The results of an evaluation of the cost effectiveness of the 1983 stream-gaging program in Kansas are documented. Data uses and funding sources were identified for the 140 complete record streamflow-gaging stations operated in Kansas during 1983 with a budget of $793,780. As a result of the evaluation of the needs and uses of data from the stream-gaging program, it was found that the 140 gaging stations were needed to meet these data requirements. The average standard error of estimation of streamflow records was 20.8 percent, assuming the 1983 budget and operating schedule of 6-week interval visitations and based on 85 of the 140 stations. It was shown that this overall level of accuracy could be improved to 18.9 percent by altering the 1983 schedule of station visitations. A minimum budget of $760 ,000, with a corresponding average error of estimation of 24.9 percent, is required to operate the 1983 program. None of the stations investigated were suitable for the application of alternative methods for simulating discharge records. Improved instrumentation can have a very positive impact on streamflow uncertainties by decreasing lost record. (USGS)

A New Correction Technique for Strain-Gage Measurements Acquired in Transient-Temperature Environments

NASA Technical Reports Server (NTRS)

Richards, W. Lance

1996-01-01

Significant strain-gage errors may exist in measurements acquired in transient-temperature environments if conventional correction methods are applied. As heating or cooling rates increase, temperature gradients between the strain-gage sensor and substrate surface increase proportionally. These temperature gradients introduce strain-measurement errors that are currently neglected in both conventional strain-correction theory and practice. Therefore, the conventional correction theory has been modified to account for these errors. A new experimental method has been developed to correct strain-gage measurements acquired in environments experiencing significant temperature transients. The new correction technique has been demonstrated through a series of tests in which strain measurements were acquired for temperature-rise rates ranging from 1 to greater than 100 degrees F/sec. Strain-gage data from these tests have been corrected with both the new and conventional methods and then compared with an analysis. Results show that, for temperature-rise rates greater than 10 degrees F/sec, the strain measurements corrected with the conventional technique produced strain errors that deviated from analysis by as much as 45 percent, whereas results corrected with the new technique were in good agreement with analytical results.

Expression, Delivery and Function of Insecticidal Proteins Expressed by Recombinant Baculoviruses

PubMed Central

Kroemer, Jeremy A.; Bonning, Bryony C.; Harrison, Robert L.

2015-01-01

Since the development of methods for inserting and expressing genes in baculoviruses, a line of research has focused on developing recombinant baculoviruses that express insecticidal peptides and proteins. These recombinant viruses have been engineered with the goal of improving their pesticidal potential by shortening the time required for infection to kill or incapacitate insect pests and reducing the quantity of crop damage as a consequence. A wide variety of neurotoxic peptides, proteins that regulate insect physiology, degradative enzymes, and other potentially insecticidal proteins have been evaluated for their capacity to reduce the survival time of baculovirus-infected lepidopteran host larvae. Researchers have investigated the factors involved in the efficient expression and delivery of baculovirus-encoded insecticidal peptides and proteins, with much effort dedicated to identifying ideal promoters for driving transcription and signal peptides that mediate secretion of the expressed target protein. Other factors, particularly translational efficiency of transcripts derived from recombinant insecticidal genes and post-translational folding and processing of insecticidal proteins, remain relatively unexplored. The discovery of RNA interference as a gene-specific regulation mechanism offers a new approach for improvement of baculovirus biopesticidal efficacy through genetic modification. PMID:25609310

Expression, delivery and function of insecticidal proteins expressed by recombinant baculoviruses.

PubMed

Kroemer, Jeremy A; Bonning, Bryony C; Harrison, Robert L

2015-01-21

Since the development of methods for inserting and expressing genes in baculoviruses, a line of research has focused on developing recombinant baculoviruses that express insecticidal peptides and proteins. These recombinant viruses have been engineered with the goal of improving their pesticidal potential by shortening the time required for infection to kill or incapacitate insect pests and reducing the quantity of crop damage as a consequence. A wide variety of neurotoxic peptides, proteins that regulate insect physiology, degradative enzymes, and other potentially insecticidal proteins have been evaluated for their capacity to reduce the survival time of baculovirus-infected lepidopteran host larvae. Researchers have investigated the factors involved in the efficient expression and delivery of baculovirus-encoded insecticidal peptides and proteins, with much effort dedicated to identifying ideal promoters for driving transcription and signal peptides that mediate secretion of the expressed target protein. Other factors, particularly translational efficiency of transcripts derived from recombinant insecticidal genes and post-translational folding and processing of insecticidal proteins, remain relatively unexplored. The discovery of RNA interference as a gene-specific regulation mechanism offers a new approach for improvement of baculovirus biopesticidal efficacy through genetic modification.

49 CFR 179.400-19 - Valves and gages.

Code of Federal Regulations, 2011 CFR

2011-10-01

... Transportation Other Regulations Relating to Transportation (Continued) PIPELINE AND HAZARDOUS MATERIALS SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) SPECIFICATIONS FOR TANK CARS Specification for Cryogenic Liquid Tank Car Tanks and Seamless Steel Tanks (Classes DOT-113 and 107A) § 179.400-19 Valves and gages...

Use of Precious Metal-Modifed Nickel-Base Superalloys for Thin Gage Applications (Preprint)

DTIC Science & Technology

2011-04-01

superalloys are being investigated for use in thin gage applications, such as thermal protection systems or heat exchangers, due to their strength and...atomic % total) in place of the platinum and iridium. 15. SUBJECT TERMS thermal protection systems, nickel, superalloy, thermomechanical processing...use in thin gage applications, such as thermal protection systems or heat exchangers, due to their strength and inherent oxidation resistance at

Exocyst Complex Protein Expression in the Human Placenta

PubMed Central

Gonzalez, I.M.; Ackerman, W.E.; Vandre, D.D.; Robinson, J.M.

2014-01-01

Introduction Protein production and secretion are essential to syncytiotrophoblast function and are associated with cytotrophoblast cell fusion and differentiation. Syncytiotrophoblast hormone secretion is a crucial determinant of maternal-fetal health, and can be misregulated in pathological pregnancies. Although, polarized secretion is a key component of placental function, the mechanisms underlying this process are poorly understood. Objective While the octameric exocyst complex is classically regarded as a master regulator of secretion in various mammalian systems, its expression in the placenta remained unexplored. We hypothesized that the syncytiotrophoblast would express all exocyst complex components and effector proteins requisite for vesicle-mediated secretion more abundantly than cytotrophoblasts in tissue specimens. Methods A two-tiered immunobiological approach was utilized to characterize exocyst and ancillary proteins in normal, term human placentas. Exocyst protein expression and localization was documented in tissue homogenates via immunoblotting and immunofluorescence labeling of placental sections. Results The eight exocyst proteins, EXOC1, 2, 3, 4, 5, 6, 7, and 8, were found in the human placenta. In addition, RAB11, an important exocyst complex modulator, was also expressed. Exocyst and Rab protein expression appeared to be regulated during trophoblast differentiation, as the syncytiotrophoblast expressed these proteins with little, if any, expression in cytotrophoblast cells. Additionally, exocyst proteins were localized at or near the syncytiotrophoblast apical membrane, the major site of placental secretion Discussion/Conclusion Our findings highlight exocyst protein expression as novel indicators of trophoblast differentiation. The exocyst’s regulated localization within the syncytiotrophoblast in conjunction with its well known functions suggests a possible role in placental polarized secretion PMID:24856041

Improving membrane protein expression by optimizing integration efficiency

PubMed Central

2017-01-01

The heterologous overexpression of integral membrane proteins in Escherichia coli often yields insufficient quantities of purifiable protein for applications of interest. The current study leverages a recently demonstrated link between co-translational membrane integration efficiency and protein expression levels to predict protein sequence modifications that improve expression. Membrane integration efficiencies, obtained using a coarse-grained simulation approach, robustly predicted effects on expression of the integral membrane protein TatC for a set of 140 sequence modifications, including loop-swap chimeras and single-residue mutations distributed throughout the protein sequence. Mutations that improve simulated integration efficiency were 4-fold enriched with respect to improved experimentally observed expression levels. Furthermore, the effects of double mutations on both simulated integration efficiency and experimentally observed expression levels were cumulative and largely independent, suggesting that multiple mutations can be introduced to yield higher levels of purifiable protein. This work provides a foundation for a general method for the rational overexpression of integral membrane proteins based on computationally simulated membrane integration efficiencies. PMID:28918393

Cost-effectiveness of the stream-gaging program in Maryland, Delaware, and the District of Columbia

USGS Publications Warehouse

Carpenter, David H.; James, R.W.; Gillen, D.F.

1987-01-01

This report documents the results of a cost-effectiveness study of the stream-gaging program in Maryland, Delaware, and the District of Columbia. Data uses and funding sources were identified for 99 continuously operated stream gages in Maryland , Delaware, and the District of Columbia. The current operation of the program requires a budget of $465,260/year. The average standard error of estimation of streamflow records is 11.8%. It is shown that this overall level of accuracy at the 99 sites could be maintained with a budget of $461,000, if resources were redistributed among the gages. (USGS)

USGS reservoir and lake gage network: Elevation and volumetric contents data, and their uses

USGS Publications Warehouse

Kroska, Anita C.

2014-01-01

In December of 2013, the U.S. Geological Survey (USGS) marked the 125th anniversary of the installation of its first official water level and streamflow gage, on the Rio Grande at Embudo, New Mexico. The gage was installed because it was recognized that water data were important to expanding irrigation needs. The USGS is a federal agency that provides nationally consistent and unbiased surface-water elevation and streamflow data at more than 10,000 gaging locations in the United States, about 330 of which are lakes and reservoirs (referred to hereafter as lakes) (Figure 1). The job of quantifying water resources, whether lakes, streams, or aquifers, is fundamental to proper water management and conservation of resources.

Physiographic and land cover attributes of the Puget Lowland and the active streamflow gaging network, Puget Sound Basin

USGS Publications Warehouse

Konrad, Christopher; Sevier, Maria

2014-01-01

Geospatial information for the active streamflow gaging network in the Puget Sound Basin was compiled to support regional monitoring of stormwater effects to small streams. The compilation includes drainage area boundaries and physiographic and land use attributes that affect hydrologic processes. Three types of boundaries were used to tabulate attributes: Puget Sound Watershed Characterization analysis units (AU); the drainage area of active streamflow gages; and the catchments of Regional Stream Monitoring Program (RSMP) sites. The active streamflow gaging network generally includes sites that represent the ranges of attributes for lowland AUs, although there are few sites with low elevations (less than 60 meters), low precipitation (less than 1 meter year), or high stream density (greater than 5 kilometers per square kilometers). The active streamflow gaging network can serve to provide streamflow information in some AUs and RSMP sites, particularly where the streamflow gage measures streamflow generated from a part of the AU or that drains to the RSMP site, and that part of the AU or RSMP site is a significant fraction of the drainage area of the streamgage. The maximum fraction of each AU or RSMP catchment upstream of a streamflow gage and the maximum fraction of any one gaged basin in an AU or RSMP along with corresponding codes are provided in the attribute tables.

Protein expression in Arabidopsis thaliana after chronic clinorotation

NASA Technical Reports Server (NTRS)

Piastuch, W. C.; Brown, C. S.

1995-01-01

Soluble protein expression in Arabidopsis thaliana L. (Heynh.) leaf and stem tissue was examined after chronic clinorotation. Seeds of Arabidopsis were germinated and plants grown to maturity on horizontal or vertical slow-rotating clinostats (1 rpm) or in stationary vertical control units. Total soluble proteins and in vivo-labeled soluble proteins isolated from these plants were analyzed by two-dimensional SDS PAGE and subsequent fluorography. Visual and computer analysis of the resulting protein patterns showed no significant differences in either total protein expression or in active protein synthesis between horizontal clinorotation and vertical controls in the Arabidopsis leaf and stem tissue. These results show chronic clinorotation does not cause gross changes in protein expression in Arabidopsis.

The Application of the Specific Gage Technique and Aerial Photographs in Kaskaskia River Degradation Studies

NASA Astrophysics Data System (ADS)

Du, X.

2008-12-01

The Kaskaskia River basin contains 136,000 acres of bottomland forest, the largest contiguous tract of bottomland forest remaining in the state of Illinois. Since the 1960's, the Carlyle Lake Dam impoundment and channelization activities have altered the natural hydrologic and ecological equilibrium of the Kaskaskia River. Morphological changes of the river channel have necessitated conservation and restoration efforts to create and maintain the sustainability, diversity, health, and connectivity of the river watershed. This study utilized the specific gage technique and historical aerial photographs to investigate the spatial and temporal changes of the river. Historical daily discharge and daily stage data from the Carlyle (1966 to 2002) and Venedy Station gages (1984 to 2003) were analyzed. Logs of daily discharge data were used to generate annual rating curves. The best fit equations were produced from annual rating regressions. A stage associated with a chosen reference discharge, the minimum available discharge (MAD), was calculated. A stage decreasing/increasing trend was used as a primary indicator of channel bed incision/aggradation. Pseudo specific gage analysis (PSGA) was used to model channel cross sectional geometry changes over time. PSGA applied similar procedures as compared to the specific gage technique. Instead of using the stage variable, PSGA utilized cross sectional width, cross-sectional area, mean velocity and gage height individually. At each gage, the historical change of each cross sectional parameter was plotted against the log of discharge. Ratings of specific stages, specific cross sectional width, specific depth, specific area, and specific velocity associated with the chosen discharge, MAD, were produced. The decreasing/increasing trend of each parameter mentioned above corresponded with changes of channel cross sectional geometries over time. Historical aerial photographs were also used to assess the bankfull channel width changing

Discharge ratings at gaging stations

USGS Publications Warehouse

Kennedy, E.J.

1984-01-01

A discharge rating is the relation of the discharge at a gaging station to stage and sometimes also to other variables. This chapter of 'Techniques of Water-Resources Investigations' describes the procedures commonly used to develop simple ratings where discharge is related only to stage and the most frequently encountered types of complex ratings where additional factors such as rate of change in stage, water-surface slope, or index velocity are used. Fundamental techniques of logarithmic plotting and the applications of simple storage routing to rating development are demonstrated. Computer applications, especially for handheld programmable calculators, and data handling are stressed.

Iterative Strain-Gage Balance Calibration Data Analysis for Extended Independent Variable Sets

NASA Technical Reports Server (NTRS)

Ulbrich, Norbert Manfred

2011-01-01

A new method was developed that makes it possible to use an extended set of independent calibration variables for an iterative analysis of wind tunnel strain gage balance calibration data. The new method permits the application of the iterative analysis method whenever the total number of balance loads and other independent calibration variables is greater than the total number of measured strain gage outputs. Iteration equations used by the iterative analysis method have the limitation that the number of independent and dependent variables must match. The new method circumvents this limitation. It simply adds a missing dependent variable to the original data set by using an additional independent variable also as an additional dependent variable. Then, the desired solution of the regression analysis problem can be obtained that fits each gage output as a function of both the original and additional independent calibration variables. The final regression coefficients can be converted to data reduction matrix coefficients because the missing dependent variables were added to the data set without changing the regression analysis result for each gage output. Therefore, the new method still supports the application of the two load iteration equation choices that the iterative method traditionally uses for the prediction of balance loads during a wind tunnel test. An example is discussed in the paper that illustrates the application of the new method to a realistic simulation of temperature dependent calibration data set of a six component balance.

Protein expression in Arabidopsis thaliana after chronic clinorotation

NASA Technical Reports Server (NTRS)

Piastuch, William C.; Brown, Christopher S.

1994-01-01

Soluble protein expression in Arabidopsis thaliana L. (Heynh.) leaf and stem tissue was examined after chronic clinorotation. Seeds of Arabidopsis were germinated and plants grown to maturity on horizontal or vertical slow-rotating clinostats (1 rpm) or in stationary vertical control units. Total soluble proteins and in vivo-labeled soluble proteins isolated from these plants were analyzed by two-dimensional sodium doedocyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and subsequent fluorography. Visual and computer analysis of the resulting protein patterns showed no significant differences in either total protein expression or in active protein synthesis between horizontal clinorotation and vertical controls in the Arabidopsis leaf and stem tissue. These results show chronic clinorotation does not cause gross changes in protein expression in Arabidopsis.

Thin layer asphaltic concrete density measuring using nuclear gages.

DOT National Transportation Integrated Search

1989-03-01

A Troxler 4640 thin layer nuclear gage was evaluated under field conditions to determine if it would provide improved accuracy of density measurements on asphalt overlays of 1-3/4 and 2 inches in thickness. Statistical analysis shows slightly improve...

Recombinant Expression Screening of P. aeruginosa Bacterial Inner Membrane Proteins

PubMed Central

2010-01-01

Background Transmembrane proteins (TM proteins) make up 25% of all proteins and play key roles in many diseases and normal physiological processes. However, much less is known about their structures and molecular mechanisms than for soluble proteins. Problems in expression, solubilization, purification, and crystallization cause bottlenecks in the characterization of TM proteins. This project addressed the need for improved methods for obtaining sufficient amounts of TM proteins for determining their structures and molecular mechanisms. Results Plasmid clones were obtained that encode eighty-seven transmembrane proteins with varying physical characteristics, for example, the number of predicted transmembrane helices, molecular weight, and grand average hydrophobicity (GRAVY). All the target proteins were from P. aeruginosa, a gram negative bacterial opportunistic pathogen that causes serious lung infections in people with cystic fibrosis. The relative expression levels of the transmembrane proteins were measured under several culture growth conditions. The use of E. coli strains, a T7 promoter, and a 6-histidine C-terminal affinity tag resulted in the expression of 61 out of 87 test proteins (70%). In this study, proteins with a higher grand average hydrophobicity and more transmembrane helices were expressed less well than less hydrophobic proteins with fewer transmembrane helices. Conclusions In this study, factors related to overall hydrophobicity and the number of predicted transmembrane helices correlated with the relative expression levels of the target proteins. Identifying physical characteristics that correlate with protein expression might aid in selecting the "low hanging fruit", or proteins that can be expressed to sufficient levels using an E. coli expression system. The use of other expression strategies or host species might be needed for sufficient levels of expression of transmembrane proteins with other physical characteristics. Surveys like this

Dark proteins: effect of inclusion body formation on quantification of protein expression.

PubMed

Iafolla, Marco A J; Mazumder, Mostafizur; Sardana, Vandit; Velauthapillai, Tharsan; Pannu, Karanbir; McMillen, David R

2008-09-01

Plasmid-borne gene expression systems have found wide application in the emerging fields of systems biology and synthetic biology, where plasmids are used to implement simple network architectures, either to test systems biology hypotheses about issues such as gene expression noise or as a means of exerting artificial control over a cell's dynamics. In both these cases, fluorescent proteins are commonly applied as a means of monitoring the expression of genes in the living cell, and efforts have been made to quantify protein expression levels through fluorescence intensity calibration and by monitoring the partitioning of proteins among the two daughter cells after division; such quantification is important in formulating the predictive models desired in systems and synthetic biology research. A potential pitfall of using plasmid-based gene expression systems is that the high protein levels associated with expression from plasmids can lead to the formation of inclusion bodies, insoluble aggregates of misfolded, nonfunctional proteins that will not generate fluorescence output; proteins caught in these inclusion bodies are thus "dark" to fluorescence-based detection methods. If significant numbers of proteins are incorporated into inclusion bodies rather than becoming biologically active, quantitative results obtained by fluorescent measurements will be skewed; we investigate this phenomenon here. We have created two plasmid constructs with differing average copy numbers, both incorporating an unregulated promoter (P(LtetO-1) in the absence of TetR) expressing the GFP derivative enhanced green fluorescent protein (EGFP), and inserted them into Escherichia coli bacterial cells (a common model organism for work on the dynamics of prokaryotic gene expression). We extracted the inclusion bodies, denatured them, and refolded them to render them active, obtaining a measurement of the average number of EGFP per cell locked into these aggregates; at the same time, we used

Evaluation of GE-167 Silicone Rubber (RTV) For Possible Service As A Moisture-Barrier For Certain Strain Gage Applications

NASA Technical Reports Server (NTRS)

Hare, David A.; Moore, Thomas C., Sr.

2000-01-01

The Langley Research Center uses strain gages in a wide variety of demanding test environments. Strain gage installations, depending on the testing scenario, may see high temperatures, cryogenic temperature, moisture accumulation, mechanical abuse, or any combination of these conditions. At Langley, there is often a need to provide protection for strain gages against moisture and mechanical abuse, especially when large-scale, harsh environment testing is to be encountered. This technical memorandum discusses the evaluation of a room temperature curing silicone rubber sealant manufactured by the General Electric Company for consideration as a moisture-barrier for certain strain gage installations.

Effects of immunosuppressive treatment on protein expression in rat kidney

PubMed Central

Kędzierska, Karolina; Sporniak-Tutak, Katarzyna; Sindrewicz, Krzysztof; Bober, Joanna; Domański, Leszek; Parafiniuk, Mirosław; Urasińska, Elżbieta; Ciechanowicz, Andrzej; Domański, Maciej; Smektała, Tomasz; Masiuk, Marek; Skrzypczak, Wiesław; Ożgo, Małgorzata; Kabat-Koperska, Joanna; Ciechanowski, Kazimierz

2014-01-01

The structural proteins of renal tubular epithelial cells may become a target for the toxic metabolites of immunosuppressants. These metabolites can modify the properties of the proteins, thereby affecting cell function, which is a possible explanation for the mechanism of immunosuppressive agents’ toxicity. In our study, we evaluated the effect of two immunosuppressive strategies on protein expression in the kidneys of Wistar rats. Fragments of the rat kidneys were homogenized after cooling in liquid nitrogen and then dissolved in lysis buffer. The protein concentration in the samples was determined using a protein assay kit, and the proteins were separated by two-dimensional electrophoresis. The obtained gels were then stained with Coomassie Brilliant Blue, and their images were analyzed to evaluate differences in protein expression. Identification of selected proteins was then performed using mass spectrometry. We found that the immunosuppressive drugs used in popular regimens induce a series of changes in protein expression in target organs. The expression of proteins involved in drug, glucose, amino acid, and lipid metabolism was pronounced. However, to a lesser extent, we also observed changes in nuclear, structural, and transport proteins’ synthesis. Very slight differences were observed between the group receiving cyclosporine, mycophenolate mofetil, and glucocorticoids (CMG) and the control group. In contrast, compared to the control group, animals receiving tacrolimus, mycophenolate mofetil, and glucocorticoids (TMG) exhibited higher expression of proteins responsible for renal drug metabolism and lower expression levels of cytoplasmic actin and the major urinary protein. In the TMG group, we observed higher expression of proteins responsible for drug metabolism and a decrease in the expression of respiratory chain enzymes (thioredoxin-2) and markers of distal renal tubular damage (heart fatty acid-binding protein) compared to expression in the CMG

Wire-Strain-Gage Hinge-Moment Indicators for Use in Tests of Airplane Models

NASA Technical Reports Server (NTRS)

Edwards, Howard B.

1944-01-01

The design and construction of various forms of strain-gage spring units and hinge-moment assemblies are discussed with particular reference to wind-tunnel test, although the indicators may be used equally well in flight tests. Strain-gage specifications are given, and the techniques of their application and use are described briefly. Testing, calibration and operation of hinge-moment indicators are discussed and precautions necessary for successful operation are stressed. Difficulties that may be encountered are summarized along with the possible causes.

Comparison of NEXRAD multisensor precipitation estimates to rain gage observations in and near DuPage County, Illinois, 2002–12

USGS Publications Warehouse

Spies, Ryan R.; Over, Thomas M.; Ortel, Terry W.

2018-05-21

In this report, precipitation data from 2002 to 2012 from the hourly gridded Next-Generation Radar (NEXRAD)-based Multisensor Precipitation Estimate (MPE) precipitation product are compared to precipitation data from two rain gage networks—an automated tipping bucket network of 25 rain gages operated by the U.S. Geological Survey (USGS) and 51 rain gages from the volunteer-operated Community Collaborative Rain, Hail, and Snow (CoCoRaHS) network—in and near DuPage County, Illinois, at a daily time step to test for long-term differences in space, time, and distribution. The NEXRAD–MPE data that are used are from the fifty 2.5-mile grid cells overlying the rain gages from the other networks. Because of the challenges of measuring of frozen precipitation, the analysis period is separated between days with or without the chance of freezing conditions. The NEXRAD–MPE and tipping-bucket rain gage precipitation data are adjusted to account for undercatch by multiplying by a previously determined factor of 1.14. Under nonfreezing conditions, the three precipitation datasets are broadly similar in cumulative depth and distribution of daily values when the data are combined spatially across the networks. However, the NEXRAD–MPE data indicate a significant trend relative to both rain gage networks as a function of distance from the NEXRAD radar just south of the study area. During freezing conditions, of the USGS network rain gages only the heated gages were considered, and these gages indicate substantial mean undercatch of 50 and 61 percent compared to the NEXRAD–MPE and the CoCoRaHS gages, respectively. The heated USGS rain gages also indicate substantially lower quantile values during freezing conditions, except during the most extreme (highest) events. Because NEXRAD precipitation products are continually evolving, the report concludes with a discussion of recent changes in those products and their potential for improved precipitation estimation. An appendix

Differential Protein Expressions in Virus-Infected and Uninfected Trichomonas vaginalis.

PubMed

He, Ding; Pengtao, Gong; Ju, Yang; Jianhua, Li; He, Li; Guocai, Zhang; Xichen, Zhang

2017-04-01

Protozoan viruses may influence the function and pathogenicity of the protozoa. Trichomonas vaginalis is a parasitic protozoan that could contain a double stranded RNA (dsRNA) virus, T. vaginalis virus (TVV). However, there are few reports on the properties of the virus. To further determine variations in protein expression of T. vaginalis , we detected 2 strains of T. vaginalis ; the virus-infected (V + ) and uninfected (V - ) isolates to examine differentially expressed proteins upon TVV infection. Using a stable isotope N-terminal labeling strategy (iTRAQ) on soluble fractions to analyze proteomes, we identified 293 proteins, of which 50 were altered in V + compared with V - isolates. The results showed that the expression of 29 proteins was increased, and 21 proteins decreased in V + isolates. These differentially expressed proteins can be classified into 4 categories: ribosomal proteins, metabolic enzymes, heat shock proteins, and putative uncharacterized proteins. Quantitative PCR was used to detect 4 metabolic processes proteins: glycogen phosphorylase, malate dehydrogenase, triosephosphate isomerase, and glucose-6-phosphate isomerase, which were differentially expressed in V + and V - isolates. Our findings suggest that mRNA levels of these genes were consistent with protein expression levels. This study was the first which analyzed protein expression variations upon TVV infection. These observations will provide a basis for future studies concerning the possible roles of these proteins in host-parasite interactions.

Analysis of D’Alembert Unfolding Technique for Hopkinson Bar Gage Records

DTIC Science & Technology

1992-06-01

MIPH No. 90652 The lfnorcqs in ttn:s ýprrt 7-n Deoarirne~nt o ý te Ip~ A~ r chn ;r vF’ pl J<P D lne! I~rozgj~lj~ The ccnitpnts of trhi-, repoit w;’ rf...0 -I0u’ -4 S.2 ,i i I II III i 00 0 04 ( d )3dflSS8dd 4 pulse is applied. This pressure pulse propagates down...GAGE WAVE TRAVEL WHEN STRESS WAVE IS AT WAV TAVE POSITIONS A, B, C, AND D Figure 2. Description of wave propagation and strain gage output in a bar

High temperature strain gages

NASA Technical Reports Server (NTRS)

Gregory, Otto J. (Inventor); You, Tao (Inventor)

2011-01-01

A ceramic strain gage based on reactively sputtered indium-tin-oxide (ITO) thin films is used to monitor the structural integrity of components employed in aerospace propulsion systems operating at temperatures in excess of 1500.degree. C. A scanning electron microscopy (SEM) of the thick ITO sensors reveals a partially sintered microstructure comprising a contiguous network of submicron ITO particles with well defined necks and isolated nanoporosity. Densification of the ITO particles was retarded during high temperature exposure with nitrogen thus stabilizing the nanoporosity. ITO strain sensors were prepared by reactive sputtering in various nitrogen/oxygen/argon partial pressures to incorporate more nitrogen into the films. Under these conditions, sintering and densification of the ITO particles containing these nitrogen rich grain boundaries was retarded and a contiguous network of nano-sized ITO particles was established.

42 CFR 84.82 - Gas pressure gages; minimum requirements.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 42 Public Health 1 2012-10-01 2012-10-01 false Gas pressure gages; minimum requirements. 84.82 Section 84.82 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES OCCUPATIONAL SAFETY AND HEALTH RESEARCH AND RELATED ACTIVITIES APPROVAL OF RESPIRATORY PROTECTIVE DEVICES Self...

42 CFR 84.82 - Gas pressure gages; minimum requirements.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 42 Public Health 1 2014-10-01 2014-10-01 false Gas pressure gages; minimum requirements. 84.82 Section 84.82 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES OCCUPATIONAL SAFETY AND HEALTH RESEARCH AND RELATED ACTIVITIES APPROVAL OF RESPIRATORY PROTECTIVE DEVICES Self...

42 CFR 84.82 - Gas pressure gages; minimum requirements.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 1 2010-10-01 2010-10-01 false Gas pressure gages; minimum requirements. 84.82 Section 84.82 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES OCCUPATIONAL SAFETY AND HEALTH RESEARCH AND RELATED ACTIVITIES APPROVAL OF RESPIRATORY PROTECTIVE DEVICES Self...

42 CFR 84.82 - Gas pressure gages; minimum requirements.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 42 Public Health 1 2013-10-01 2013-10-01 false Gas pressure gages; minimum requirements. 84.82 Section 84.82 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES OCCUPATIONAL SAFETY AND HEALTH RESEARCH AND RELATED ACTIVITIES APPROVAL OF RESPIRATORY PROTECTIVE DEVICES Self...

42 CFR 84.82 - Gas pressure gages; minimum requirements.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 1 2011-10-01 2011-10-01 false Gas pressure gages; minimum requirements. 84.82 Section 84.82 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES OCCUPATIONAL SAFETY AND HEALTH RESEARCH AND RELATED ACTIVITIES APPROVAL OF RESPIRATORY PROTECTIVE DEVICES Self...

Design of a Multifrequency Reed-Type Shock Gage With an Inertia-Operated Trigger

DTIC Science & Technology

1949-05-01

SSBärl=^isä&^^^^^^^Z^^^mSSSn^ •RAPE£LHQLDIUG PLATE.- SEED CDiÜMM_SKAE PÄPEP"C*£I:AMP^-, / SHAFTS SPACEß COMPEESSION OI?WE SPEIW PAPEB HOIOEE LATCH ...which wedges it against a sta- tionary plate located opposite the ratchet. Operation of Gage After the gage is mounted on the structure to be...clamping nut, waxed paper is inserted, and the paper holder is pulled back 2 in. by hand until it is engaged by a paper-holder latch provided at the

Instrumentation, methods of flood-data collection and transmission, and evaluation of streamflow-gaging network in Indiana

USGS Publications Warehouse

Glatfelter, D.R.; Butch, G.K.

1994-01-01

The study results indicate that installation of streamflow-gaging stations at 15 new sites would improve collection of flood data. Instrumenting the 15 new sites plus 26 existing streamflow-gaging stations with telemetry, preferably data-collection platforms with satellite transmitters, would improve transmission of data to users of the information.

Electrical Properties of Materials for Elevated Temperature Resistance Strain Gage Application. Ph.D. Thesis

NASA Technical Reports Server (NTRS)

Lei, Jih-Fen

1987-01-01

The objective was to study the electrical resistances of materials that are potentially useful as resistance strain gages at 1000 C. Transition metal carbides and nitrides, boron carbide and silicon carbide were selected for the experimental phase of this research. Due to their low temperature coefficient of resistance and good stability, TiC, ZrC, B sub 4 C and beta-SiC are suggested as good candidates for high temperature resistance strain gage applications.

Expression of multidrug resistance proteins in retinoblastoma

PubMed Central

Shukla, Swati; Srivastava, Arpna; Kumar, Sunil; Singh, Usha; Goswami, Sandeep; Chawla, Bhavna; Bajaj, Mandeep Singh; Kashyap, Seema; Kaur, Jasbir

2017-01-01

AIM To elucidate the mechanism of multidrug resistance in retinoblastoma, and to acquire more insights into in vivo drug resistance. METHODS Three anticancer drug resistant Y79 human RB cells were generated against vincristine, etoposide or carboplatin, which are used for conventional chemotherapy in RB. Primary cultures from enucleated eyes after chemotherapy (PCNC) were also prepared. Their chemosensitivity to chemotherapeutic agents (vincristine, etoposide and carboplatin) were measured using MTT assay. Western blot analysis was performed to evaluate the expression of p53, Bcl-2 and various multidrug resistant proteins in retinoblastoma cells. RESULTS Following exposure to chemotherapeutic drugs, PCNC showed less sensitivity to drugs. No significant changes observed in the p53 expression, whereas Bcl-2 expression was found to be increased in the drug resistant cells as well as in PCNC. Increased expression of P-glycoprotein (P-gp) was observed in drug resistant Y79 cells; however there was no significant change in the expression of P-gp found between primary cultures of primarily enucleated eyes and PCNC. Multidrug resistance protein 1 (Mrp-1) expression was found to be elevated in the drug resistant Y79 cells as well as in PCNC. No significant change in the expression of lung resistance associated protein (Lrp) was observed in the drug resistant Y79 cells as well as in PCNC. CONCLUSION Our results suggest that multidrug resistant proteins are intrinsically present in retinoblastoma which causes treatment failure in managing retinoblastoma with chemotherapy. PMID:29181307

Nucleic acid programmable protein array a just-in-time multiplexed protein expression and purification platform.

PubMed

Qiu, Ji; LaBaer, Joshua

2011-01-01

Systematic study of proteins requires the availability of thousands of proteins in functional format. However, traditional recombinant protein expression and purification methods have many drawbacks for such study at the proteome level. We have developed an innovative in situ protein expression and capture system, namely NAPPA (nucleic acid programmable protein array), where C-terminal tagged proteins are expressed using an in vitro expression system and efficiently captured/purified by antitag antibodies coprinted at each spot. The NAPPA technology presented in this chapter enable researchers to produce and display fresh proteins just in time in a multiplexed high-throughput fashion and utilize them for various downstream biochemical researches of interest. This platform could revolutionize the field of functional proteomics with it ability to produce thousands of spatially separated proteins in high density with narrow dynamic rand of protein concentrations, reproducibly and functionally. Copyright © 2011 Elsevier Inc. All rights reserved.

50. Stream gaging station in steelpipe well and shelter, looking ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

50. Stream gaging station in steel-pipe well and shelter, looking west. Photo by Robin Lee Tedder, Puget Power, 1989. - Puget Sound Power & Light Company, White River Hydroelectric Project, 600 North River Avenue, Dieringer, Pierce County, WA

26. Mechanical float gages used to monitor level of water ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

26. Mechanical float gages used to monitor level of water in the filtration bed reservoir. - Lake Whitney Water Filtration Plant, Filtration Plant, South side of Armory Street between Edgehill Road & Whitney Avenue, Hamden, New Haven County, CT

High Temperature - Thin Film Strain Gages Based on Alloys of Indium Tin Oxide

NASA Technical Reports Server (NTRS)

Gregory, Otto J.; Cooke, James D.; Bienkiewicz, Joseph M.

1998-01-01

A stable, high temperature strain gage based on reactively sputtered indium tin oxide (ITO) was demonstrated at temperatures up to 1050 C. These strain sensors exhibited relatively large, negative gage factors at room temperature and their piezoresistive response was both linear and reproducible when strained up to 700 micro-in/in. When cycled between compression and tension, these sensors also showed very little hysteresis, indicating excellent mechanical stability. Thin film strain gages based on selected ITO alloys withstood more than 50,000 strain cycles of +/- 500 micro-in/in during 180 hours of testing in air at 1000 C, with minimal drift at temperature. Drift rates as low as 0.0009%/hr at 1000 C were observed for ITO films that were annealed in nitrogen at 700 C prior to strain testing. These results compare favorably with state of the art 10 micro-m thick PdCr films deposited by NASA, where drift rates of 0.047%/hr at 1050 C were observed. Nitrogen annealing not only produced the lowest drift rates to date, but also produce the largest dynamic gage factors (G = 23.5). These wide bandgap, semiconductor strain sensors also exhibited moderately low temperature coefficients of resistance (TCR) at temperatures up to 1100 C, when tested in a nitrogen ambient. A TCR of +230 ppm/C over the temperature range 200 C < T < 500 C and a TCR of -469 ppm/C over the temperature range 600 C < T < 1100 C was observed for the films tested in nitrogen. However, the resistivity behavior changed considerably when the same films were tested in oxygen ambients. A TCR of -1560 ppm/C was obtained over the temperature range of 200 C < T < 1100 C. When similar films were protected with an overcoat or when ITO films were prepared with higher oxygen contents in the plasma, two distinct TCR's were observed. At T < 800 C, a linear TCR of -210 ppm/C was observed and at T > 800 C, a linear TCR of -2170 DDm/C was observed. The combination of a moderately low TCR and a relatively large gage

46 CFR 98.25-45 - Liquid level gaging device.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 46 Shipping 4 2011-10-01 2011-10-01 false Liquid level gaging device. 98.25-45 Section 98.25-45 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS SPECIAL CONSTRUCTION, ARRANGEMENT, AND OTHER PROVISIONS FOR CERTAIN DANGEROUS CARGOES IN BULK Anhydrous Ammonia in Bulk...

46 CFR 98.25-45 - Liquid level gaging device.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 46 Shipping 4 2013-10-01 2013-10-01 false Liquid level gaging device. 98.25-45 Section 98.25-45 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS SPECIAL CONSTRUCTION, ARRANGEMENT, AND OTHER PROVISIONS FOR CERTAIN DANGEROUS CARGOES IN BULK Anhydrous Ammonia in Bulk...

46 CFR 98.25-45 - Liquid level gaging device.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 46 Shipping 4 2014-10-01 2014-10-01 false Liquid level gaging device. 98.25-45 Section 98.25-45 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS SPECIAL CONSTRUCTION, ARRANGEMENT, AND OTHER PROVISIONS FOR CERTAIN DANGEROUS CARGOES IN BULK Anhydrous Ammonia in Bulk...

46 CFR 98.25-45 - Liquid level gaging device.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 46 Shipping 4 2010-10-01 2010-10-01 false Liquid level gaging device. 98.25-45 Section 98.25-45 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS SPECIAL CONSTRUCTION, ARRANGEMENT, AND OTHER PROVISIONS FOR CERTAIN DANGEROUS CARGOES IN BULK Anhydrous Ammonia in Bulk...

46 CFR 98.25-45 - Liquid level gaging device.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 46 Shipping 4 2012-10-01 2012-10-01 false Liquid level gaging device. 98.25-45 Section 98.25-45 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS SPECIAL CONSTRUCTION, ARRANGEMENT, AND OTHER PROVISIONS FOR CERTAIN DANGEROUS CARGOES IN BULK Anhydrous Ammonia in Bulk...

49. View of unlined canal near inline stream gaging station, ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

49. View of unlined canal near in-line stream gaging station, looking west. Photo by Robin Lee Tedder, Puget Power, 1989. - Puget Sound Power & Light Company, White River Hydroelectric Project, 600 North River Avenue, Dieringer, Pierce County, WA

49 CFR 213.110 - Gage restraint measurement systems.

Code of Federal Regulations, 2014 CFR

2014-10-01

... applied force from 0 to 4,000 pounds; and (ii) The gage of the track under 4,000 pounds of applied force... requirements specified in §§ 213.109 and 213.127. (5) If the PTLF becomes non-functional or is missing, the... vertical loading of the track is necessary for contact with the lateral rail restraint components, a PTLF...

49 CFR 213.110 - Gage restraint measurement systems.

Code of Federal Regulations, 2012 CFR

2012-10-01

... applied force from 0 to 4,000 pounds; and (ii) The gage of the track under 4,000 pounds of applied force... requirements specified in §§ 213.109 and 213.127. (5) If the PTLF becomes non-functional or is missing, the... vertical loading of the track is necessary for contact with the lateral rail restraint components, a PTLF...

49 CFR 213.110 - Gage restraint measurement systems.

Code of Federal Regulations, 2013 CFR

2013-10-01

... applied force from 0 to 4,000 pounds; and (ii) The gage of the track under 4,000 pounds of applied force... requirements specified in §§ 213.109 and 213.127. (5) If the PTLF becomes non-functional or is missing, the... vertical loading of the track is necessary for contact with the lateral rail restraint components, a PTLF...

Advances in recombinant protein expression for use in pharmaceutical research.

PubMed

Assenberg, Rene; Wan, Paul T; Geisse, Sabine; Mayr, Lorenz M

2013-06-01

Protein production for structural and biophysical studies, functional assays, biomarkers, mechanistic studies in vitro and in vivo, but also for therapeutic applications in pharma, biotech and academia has evolved into a mature discipline in recent years. Due to the increased emphasis on biopharmaceuticals, the growing demand for proteins used for structural and biophysical studies, the impact of genomics technologies on the analysis of large sets of structurally diverse proteins, and the increasing complexity of disease targets, the interest in innovative approaches for the expression, purification and characterisation of recombinant proteins has steadily increased over the years. In this review, we summarise recent developments in the field of recombinant protein expression for research use in pharma, biotech and academia. We focus mostly on the latest developments for protein expression in the most widely used expression systems: Escherichia coli (E. coli), insect cell expression using the Baculovirus Expression Vector System (BEVS) and, finally, transient and stable expression of recombinant proteins in mammalian cells. Copyright © 2013. Published by Elsevier Ltd.

Using Peptide-Level Proteomics Data for Detecting Differentially Expressed Proteins.

PubMed

Suomi, Tomi; Corthals, Garry L; Nevalainen, Olli S; Elo, Laura L

2015-11-06

The expression of proteins can be quantified in high-throughput means using different types of mass spectrometers. In recent years, there have emerged label-free methods for determining protein abundance. Although the expression is initially measured at the peptide level, a common approach is to combine the peptide-level measurements into protein-level values before differential expression analysis. However, this simple combination is prone to inconsistencies between peptides and may lose valuable information. To this end, we introduce here a method for detecting differentially expressed proteins by combining peptide-level expression-change statistics. Using controlled spike-in experiments, we show that the approach of averaging peptide-level expression changes yields more accurate lists of differentially expressed proteins than does the conventional protein-level approach. This is particularly true when there are only few replicate samples or the differences between the sample groups are small. The proposed technique is implemented in the Bioconductor package PECA, and it can be downloaded from http://www.bioconductor.org.

Importance of understanding landscape biases in USGS gage locations: Implications and solutions for managers

USGS Publications Warehouse

Wagner, Tyler; DeWeber, Jefferson Tyrell; Tsang, Yin-Phan; Krueger, Damon; Whittier, Joanna B.; Infante, Dana M.; Whelan, Gary

2014-01-01

Flow and water temperature are fundamental properties of stream ecosystems upon which many freshwater resource management decisions are based. U.S. Geological Survey (USGS) gages are the most important source of streamflow and water temperature data available nationwide, but the degree to which gages represent landscape attributes of the larger population of streams has not been thoroughly evaluated. We identified substantial biases for seven landscape attributes in one or more regions across the conterminous United States. Streams with small watersheds (<10 km2) and at high elevations were often underrepresented, and biases were greater for water temperature gages and in arid regions. Biases can fundamentally alter management decisions and at a minimum this potential for error must be acknowledged accurately and transparently. We highlight three strategies that seek to reduce bias or limit errors arising from bias and illustrate how one strategy, supplementing USGS data, can greatly reduce bias.

SBDS Protein Expression Patterns in the Bone Marrow

PubMed Central

Wong, Trisha E.; Calicchio, Monica L.; Fleming, Mark D.; Shimamura, Akiko; Harris, Marian H.

2010-01-01

Shwachman Diamond Syndrome (SDS) is an inherited bone marrow failure syndrome caused by biallelic SBDS gene mutations. Here we examined SBDS protein levels in human bone marrow. SBDS protein expression was high in neutrophil progenitors, megakaryocytes, plasma cells and osteoblasts. In contrast, SBDS protein levels were low in all hematopoietic cell lineages from patients harboring the common SBDS mutations. We conclude that SBDS protein levels vary widely between specific marrow lineages. Uniformly low SBDS protein expression levels distinguish the majority of SDS patients from controls or other marrow failure syndromes. PMID:20658628

Vectors for co-expression of an unrestricted number of proteins

PubMed Central

Scheich, Christoph; Kümmel, Daniel; Soumailakakis, Dimitri; Heinemann, Udo; Büssow, Konrad

2007-01-01

A vector system is presented that allows generation of E. coli co-expression clones by a standardized, robust cloning procedure. The number of co-expressed proteins is not limited. Five ‘pQLink’ vectors for expression of His-tag and GST-tag fusion proteins as well as untagged proteins and for cloning by restriction enzymes or Gateway cloning were generated. The vectors allow proteins to be expressed individually; to achieve co-expression, two pQLink plasmids are combined by ligation-independent cloning. pQLink co-expression plasmids can accept an unrestricted number of genes. As an example, the co-expression of a heterotetrameric human transport protein particle (TRAPP) complex from a single plasmid, its isolation and analysis of its stoichiometry are shown. pQLink clones can be used directly for pull-down experiments if the proteins are expressed with different tags. We demonstrate pull-down experiments of human valosin-containing protein (VCP) with fragments of the autocrine motility factor receptor (AMFR). The cloning method avoids PCR or gel isolation of restriction fragments, and a single resistance marker and origin of replication are used, allowing over-expression of rare tRNAs from a second plasmid. It is expected that applications are not restricted to bacteria, but could include co-expression in other hosts such as Bacluovirus/insect cells. PMID:17311810

[Prokaryotic expression, purification and antigenicity identification of recombinant human survivin protein].

PubMed

Yin, Xiaotao; Wang, Wei; Tian, Renli; Xu, Yuanji; Yan, Jinqi; Zhang, Wei; Gao, Jiangping; Yu, Jiyun

2013-08-01

To construct a prokaryotic expression plasmid pET28a-survivin, optimize the recombinant protein expression conditions in E.coli, and purify the survivin recombinant protein and identify its antigenicity. Survivin cDNA segment was amplified by PCR and cloned into prokaryotic expression vector pET28a(+) to construct the recombinant expression vector pET28a-survivin. The expression vector was transformed into BL21 (DE3) and the fusion protein survivin/His was induced by IPTG. The fusion protein was purified through Ni affinity chromatography. The antigenicity of the purified survivin protein was identified by Western blotting and ELISA. The recombinant expression vector was verified successfully by BamHI and HindIII. The fusion protein induced by IPTG was obtained with Mr; about 24 000. The purity of the purified protein reached 90% by SDS-PAGE analysis. And the antigenicity of the survivin protein was validated by Western blotting and ELISA. The prokaryotic expression plasmid pET28a-survivin was successfully constructed and the survivin protein was expressed and purified in E.coli. The antigenicity of the purified survivin protein was demonstrated desirable.

The growing impact of lyophilized cell-free protein expression systems

PubMed Central

Hunt, J. Porter; Yang, Seung Ook; Wilding, Kristen M.; Bundy, Bradley C.

2017-01-01

ABSTRACT Recently reported shelf-stable, on-demand protein synthesis platforms are enabling new possibilities in biotherapeutics, biosensing, biocatalysis, and high throughput protein expression. Lyophilized cell-free protein expression systems not only overcome cold-storage limitations, but also enable stockpiling for on-demand synthesis and completely sterilize the protein synthesis platform. Recently reported high-yield synthesis of cytotoxic protein Onconase from lyophilized E. coli extract preparations demonstrates the utility of lyophilized cell-free protein expression and its potential for creating on-demand biotherapeutics, vaccines, biosensors, biocatalysts, and high throughput protein synthesis. PMID:27791452

Reliability of the Forest Service type rain gage

Treesearch

G. L. Hayes

1942-01-01

The Forest Service type rain-gage was originated by W. B. Osborne of the Portland, Oreg., office of the U. S. Forest Service to meet a need for reliable rainfall measurements at low cost. Many hundreds of them, over 700 on the national forests of northern Idaho and western Montana alone, are now being used during the summer months as aids to forest-fire control...

QNSE Theory of Turbulence in Rotating Fluids and the Nastrom & Gage Spectrum

NASA Astrophysics Data System (ADS)

Galperin, B.

2017-12-01

An analytical theory of turbulence, the quasi-normal scale elimination (QNSE), has been developed for neutrally stratified rotating flows. The theory provides near-first principle framework for the representation of flow anisotropization under the action of rotation. The anisotropization reveals itself in the emergence of different eddy viscosities and eddy diffusivities in different directions and directional dependence of the kinetic and potential energies spectra. In addition, there are also phenomena of componentality, eddy viscosities are different for different velocity components, and the onset of the inverse energy cascade. The anisotropization increases with increasing scale. The characteristic scales for the crossover between the turbulence and inertial wave domains is the Woods scale, LΩ = [ɛ/(2Ω)3)]1/2, ɛ being the rate of the viscous dissipation, which is analogous to the Ozmidov scale in flows with stable stratification. Rapid rotation renders the horizontal eddy viscosity negative, and in order to preserve it positive, a weak rotation limit is invoked. Within that limit, an analytical theory of the transition from the Kolmogorov to a rotation-dominated turbulence regime is developed. The dispersion relation of linear inertial waves is unaffected by turbulence while all one-dimensional energy spectra undergo steepening from the Kolmogorov -5/3 to the -3 slope. The longitudinal and transverse spectra are congruent with the famous atmospheric spectra by Nastrom & Gage. Thus, for the first time, these spectra are obtained within an analytical theory. QNSE explains the latitudinal dependence of the spectra and lends itself for practical applications in simulations of atmospheric and oceanic flows as it produces closed expressions for the eddy viscosities and eddy diffusivities. The Nastrom & Gage spectra also apply to the oceanic flows.

The return of Phineas Gage: clues about the brain from the skull of a famous patient.

PubMed

Damasio, H; Grabowski, T; Frank, R; Galaburda, A M; Damasio, A R

1994-05-20

When the landmark patient Phineas Gage died in 1861, no autopsy was performed, but his skull was later recovered. The brain lesion that caused the profound personality changes for which his case became famous has been presumed to have involved the left frontal region, but questions have been raised about the involvement of other regions and about the exact placement of the lesion within the vast frontal territory. Measurements from Gage's skull and modern neuroimaging techniques were used to reconstitute the accident and determine the probable location of the lesion. The damage involved both left and right prefrontal cortices in a pattern that, as confirmed by Gage's modern counterparts, causes a defect in rational decision making and the processing of emotion.

Development of lightweight graphite/polyimide sandwich panels. Phase 2: Thin gage material manufacture

NASA Technical Reports Server (NTRS)

Merlette, J. B.

1972-01-01

Thin gage materials selected and the rationale for their basic requirements are discussed. The resin used in all prepreg manufacture is Monsanto RS-6234 polyimide. The selected fiber for core manufacture is Hercules HT-S, and the selected fiber for face sheets is Hercules HM-S. The technique for making thin gage prepreg was to wind spread carbon fiber tows into a resin film on a large drum. This technique was found to be superior to others investigated. A total of 22 pounds of 1 to 2 mil/ply prepreg was fabricated for use on the program.

A Conversion of Wheatstone Bridge to Current-Loop Signal Conditioning for Strain Gages

NASA Technical Reports Server (NTRS)

Anderson, Karl F.

1995-01-01

Current loop circuitry replaced Wheatstone bridge circuitry to signal-condition strain gage transducers in more than 350 data channels for two different test programs at NASA Dryden Flight Research Center. The uncorrected test data from current loop circuitry had a lower noise level than data from comparable Wheatstone bridge circuitry, were linear with respect to gage-resistance change, and were uninfluenced by varying lead-wire resistance. The current loop channels were easier for the technicians to set up, verify, and operate than equivalent Wheatstone bridge channels. Design choices and circuit details are presented in this paper in addition to operational experience.

High temperature static strain gage development contract, tasks 1 and 2

NASA Technical Reports Server (NTRS)

Hulse, C. O.; Bailey, R. S.; Grant, H. P.; Przybyszewski, J. S.

1987-01-01

Results are presented for the first two tasks to develop resistive strain gage systems for use up to 1250 K on blades and vanes in gas turbine engines under tests. The objective of these two tasks was to further improve and evaluate two static strain gage alloys identified as candidates in a previous program. Improved compositions were not found for either alloy. Further efforts on the Fe-11.9Al-10.6Cr weigth percent alloy were discontinued because of time dependent drift problems at 1250 K in air. When produced as a 6.5 micrometer thick sputtered film, the Pd-13Cr weight percent alloys is not sufficiently stable for this use in air at 1250 K and a protective overcoat system will need to be developed.

[Expression of c-jun protein after experimental rat brain concussion].

PubMed

Wang, Feng; Li, Yong-hong

2010-02-01

To observe e-jun protein expression after rat brain concussion and explore the forensic pathologic markers following brain concussion. Fifty-five rats were randomly divided into brain concussion group and control group. The expression of c-jun protein was observed by immunohistochemistry. There were weak positive expression of c-jun protein in control group. In brain concussion group, however, some neutrons showed positive expression of c-jun protein at 15 min after brain concussion, and reach to the peak at 3 h after brain concussion. The research results suggest that detection of c-jun protein could be a marker to determine brain concussion and estimate injury time after brain concussion.

Production of therapeutic proteins in algae, analysis of expression of seven human proteins in the chloroplast of Chlamydomonas reinhardtii

PubMed Central

Rasala, Beth A; Muto, Machiko; Lee, Philip A; Jager, Michal; Cardoso, Rosa MF; Behnke, Craig A; Kirk, Peter; Hokanson, Craig A; Crea, Roberto; Mendez, Michael; Mayfield, Stephen P

2010-01-01

Summary Recombinant proteins are widely used today in many industries, including the biopharmaceutical industry, and can be expressed in bacteria, yeasts, mammalian and insect cell cultures, or in transgenic plants and animals. In addition, transgenic algae have also been shown to support recombinant protein expression, both from the nuclear and chloroplast genomes. However, to date, there are only a few reports on recombinant proteins expressed in the algal chloroplast. It is unclear if this is due to few attempts or to limitations of the system that preclude expression of many proteins. Thus, we sought to assess the versatility of transgenic algae as a recombinant protein production platform. To do this, we tested whether the algal chloroplast could support the expression of a diverse set of current or potential human therapeutic proteins. Of the seven proteins chosen, greater than 50% expressed at levels sufficient for commercial production. Three expressed at 2% to 3% of total soluble protein, while a forth protein accumulated to similar levels when translationally fused to a well-expressed serum amyloid protein. All of the algal chloroplast-expressed proteins are soluble and showed biological activity comparable to that of the same proteins expressed using traditional production platforms. Thus, the success rate, expression levels, and bioactivty achieved demonstrate the utility of C. reinhardtii as a robust platform for human therapeutic protein production. PMID:20230484

Cost-effectiveness of the U.S. Geological Survey's stream-gaging programs in Massachusetts and Rhode Island

USGS Publications Warehouse

Gadoury, R.A.; Smath, J.A.; Fontaine, R.A.

1985-01-01

The report documents the results of a study of the cost-effectiveness of the U.S. Geological Survey 's continuous-record stream-gaging programs in Massachusetts and Rhode Island. Data uses and funding sources were identified for 91 gaging stations being operated in Massachusetts are being operated to provide data for two special purpose hydrologic studies, and they are planned to be discontinued at the conclusion of the studies. Cost-effectiveness analyses were performed on 63 continuous-record gaging stations in Massachusetts and 15 stations in Rhode Island, at budgets of $353,000 and $60,500, respectively. Current operations policies result in average standard errors per station of 12.3% in Massachusetts and 9.7% in Rhode Island. Minimum possible budgets to maintain the present numbers of gaging stations in the two States are estimated to be $340,000 and $59,000, with average errors per station of 12.8% and 10.0%, respectively. If the present budget levels were doubled, average standards errors per station would decrease to 8.1% and 4.2%, respectively. Further budget increases would not improve the standard errors significantly. (USGS)

High-Throughput Protein Expression Using a Combination of Ligation-Independent Cloning (LIC) and Infrared Fluorescent Protein (IFP) Detection

PubMed Central

Dortay, Hakan; Akula, Usha Madhuri; Westphal, Christin; Sittig, Marie; Mueller-Roeber, Bernd

2011-01-01

Protein expression in heterologous hosts for functional studies is a cumbersome effort. Here, we report a superior platform for parallel protein expression in vivo and in vitro. The platform combines highly efficient ligation-independent cloning (LIC) with instantaneous detection of expressed proteins through N- or C-terminal fusions to infrared fluorescent protein (IFP). For each open reading frame, only two PCR fragments are generated (with three PCR primers) and inserted by LIC into ten expression vectors suitable for protein expression in microbial hosts, including Escherichia coli, Kluyveromyces lactis, Pichia pastoris, the protozoon Leishmania tarentolae, and an in vitro transcription/translation system. Accumulation of IFP-fusion proteins is detected by infrared imaging of living cells or crude protein extracts directly after SDS-PAGE without additional processing. We successfully employed the LIC-IFP platform for in vivo and in vitro expression of ten plant and fungal proteins, including transcription factors and enzymes. Using the IFP reporter, we additionally established facile methods for the visualisation of protein-protein interactions and the detection of DNA-transcription factor interactions in microtiter and gel-free format. We conclude that IFP represents an excellent reporter for high-throughput protein expression and analysis, which can be easily extended to numerous other expression hosts using the setup reported here. PMID:21541323

Gage restraint measurement system comparison tests : railbound and hi-rail vehicles

DOT National Transportation Integrated Search

2003-12-01

Comparative tests were conducted to evaluate the gage restraint measurement system (GRMS) testing capabilities of a railbound GRMS vehicle, Federal Railroad Administration's (FRA's) T-6, and a hi-rail vehicle, Holland Company's TrackStar GRMS system....

Heterologous expression of proteins from Plasmodium falciparum: results from 1000 genes.

PubMed

Mehlin, Christopher; Boni, Erica; Buckner, Frederick S; Engel, Linnea; Feist, Tiffany; Gelb, Michael H; Haji, Lutfiyah; Kim, David; Liu, Colleen; Mueller, Natascha; Myler, Peter J; Reddy, J T; Sampson, Joshua N; Subramanian, E; Van Voorhis, Wesley C; Worthey, Elizabeth; Zucker, Frank; Hol, Wim G J

2006-08-01

As part of a structural genomics initiative, 1000 open reading frames from Plasmodium falciparum, the causative agent of the most deadly form of malaria, were tested in an E. coli protein expression system. Three hundred and thirty-seven of these targets were observed to express, although typically the protein was insoluble. Sixty-three of the targets provided soluble protein in yields ranging from 0.9 to 406.6 mg from one liter of rich media. Higher molecular weight, greater protein disorder (segmental analysis, SEG), more basic isoelectric point (pI), and a lack of homology to E. coli proteins were all highly and independently correlated with difficulties in expression. Surprisingly, codon usage and the percentage of adenosines and thymidines (%AT) did not appear to play a significant role. Of those proteins which expressed, high pI and a hypothetical annotation were both strongly and independently correlated with insolubility. The overwhelmingly important role of pI in both expression and solubility appears to be a surprising and fundamental issue in the heterologous expression of P. falciparum proteins in E. coli. Twelve targets which did not express in E. coli from the native gene sequence were codon-optimized through whole gene synthesis, resulting in the (insoluble) expression of three of these proteins. Seventeen targets which were expressed insolubly in E. coli were moved into a baculovirus/Sf-21 system, resulting in the soluble expression of one protein at a high level and six others at a low level. A variety of factors conspire to make the heterologous expression of P. falciparum proteins challenging, and these observations lay the groundwork for a rational approach to prioritizing and, ultimately, eliminating these impediments.

[Pichia pastoris as an expression system for recombinant protein production].

PubMed

Ciarkowska, Anna; Jakubowska, Anna

2013-01-01

Pichia pastoris has become increasingly popular as a host for recombinant protein production in recent years. P. pastoris is more cost effective and allows achieving higher expression levels than insect and mammalian cells. It also offers some significant advantages over E. coli expression systems, such as avoiding problems with proper protein folding. Also, P. pastoris as an eukaryotic organism can carry out posttranslational modifications of produced proteins. Additionally, P. pastoris can produce high levels of recombinant proteins in extracellular medium which simplifies protein purification. Having many advantages over other expression systems makes P. pastoris an organism of choice for industrial protein production.

13. VIEW OF SHEFFIELD GAGE, USED TO MEASURE ROUNDNESS OF ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

13. VIEW OF SHEFFIELD GAGE, USED TO MEASURE ROUNDNESS OF WEAPON PARTS, AS PART OF THE FINAL PARTS INSPECTION PROCESS. (6/2/67) - Rocky Flats Plant, Non-Nuclear Production Facility, South of Cottonwood Avenue, west of Seventh Avenue & east of Building 460, Golden, Jefferson County, CO

Suitability of Strain Gage Sensors for Integration into Smart Sport Equipment: A Golf Club Example.

PubMed

Umek, Anton; Zhang, Yuan; Tomažič, Sašo; Kos, Anton

2017-04-21

Wearable devices and smart sport equipment are being increasingly used in amateur and professional sports. Smart sport equipment employs various sensors for detecting its state and actions. The correct choice of the most appropriate sensor(s) is of paramount importance for efficient and successful operation of sport equipment. When integrated into the sport equipment, ideal sensors are unobstructive, and do not change the functionality of the equipment. The article focuses on experiments for identification and selection of sensors that are suitable for the integration into a golf club with the final goal of their use in real time biofeedback applications. We tested two orthogonally affixed strain gage (SG) sensors, a 3-axis accelerometer, and a 3-axis gyroscope. The strain gage sensors are calibrated and validated in the laboratory environment by a highly accurate Qualisys Track Manager (QTM) optical tracking system. Field test results show that different types of golf swing and improper movement in early phases of golf swing can be detected with strain gage sensors attached to the shaft of the golf club. Thus they are suitable for biofeedback applications to help golfers to learn repetitive golf swings. It is suggested that the use of strain gage sensors can improve the golf swing technical error detection accuracy and that strain gage sensors alone are enough for basic golf swing analysis. Our final goal is to be able to acquire and analyze as many parameters of a smart golf club in real time during the entire duration of the swing. This would give us the ability to design mobile and cloud biofeedback applications with terminal or concurrent feedback that will enable us to speed-up motor skill learning in golf.

Suitability of Strain Gage Sensors for Integration into Smart Sport Equipment: A Golf Club Example

PubMed Central

Umek, Anton; Zhang, Yuan; Tomažič, Sašo; Kos, Anton

2017-01-01

Wearable devices and smart sport equipment are being increasingly used in amateur and professional sports. Smart sport equipment employs various sensors for detecting its state and actions. The correct choice of the most appropriate sensor(s) is of paramount importance for efficient and successful operation of sport equipment. When integrated into the sport equipment, ideal sensors are unobstructive, and do not change the functionality of the equipment. The article focuses on experiments for identification and selection of sensors that are suitable for the integration into a golf club with the final goal of their use in real time biofeedback applications. We tested two orthogonally affixed strain gage (SG) sensors, a 3-axis accelerometer, and a 3-axis gyroscope. The strain gage sensors are calibrated and validated in the laboratory environment by a highly accurate Qualisys Track Manager (QTM) optical tracking system. Field test results show that different types of golf swing and improper movement in early phases of golf swing can be detected with strain gage sensors attached to the shaft of the golf club. Thus they are suitable for biofeedback applications to help golfers to learn repetitive golf swings. It is suggested that the use of strain gage sensors can improve the golf swing technical error detection accuracy and that strain gage sensors alone are enough for basic golf swing analysis. Our final goal is to be able to acquire and analyze as many parameters of a smart golf club in real time during the entire duration of the swing. This would give us the ability to design mobile and cloud biofeedback applications with terminal or concurrent feedback that will enable us to speed-up motor skill learning in golf. PMID:28430147

[Expression of Dengue virus type 2 nonstructural protein 3 and isolation of host proteins interacting with it].

PubMed

Weng, Daihui; Lei, Yingfeng; Dong, Yangchao; Han, Peijun; Ye, Chuantao; Yang, Jing; Wang, Yuan; Yin, Wen

2015-12-01

To construct the plasmid expressing the fusion protein of Dengue virus type 2 (DENV2) nonstructural protein 3 (NS3) with affinity tag, and isolate the cellular proteins interacting with NS3 protein using tandem affinity purification (TAP) assay. Primers for amplifying NS3 gene were designed according to the sequence of DENV2 genome and chemically synthesized. The NS3 fragments, after amplified by PCR with DENV2 cDNA as template, were digested and cloned into the mammalian eukaryotic expression vector pCI-SF with the tandem affinity tag (FLAG-StrepII). The recombinant pCI-NS3-SF was transiently transformed by Lipofectamine(TM) 2000 into HEK293T cells, and the expression of the fusion protein was confirmed by Western blotting. Cellular proteins that interacted with NS3 were isolated and purified by TAP assay. The eukaryotic expression vector expressing NS3 protein was successfully constructed. The host proteins interacting with NS3 protein were isolated by TAP system. TAP is an efficient method to isolate the cellular proteins interacting with DENV2 NS3.

Biotechnology Protein Expression and Purification Facility

NASA Technical Reports Server (NTRS)

2003-01-01

The purpose of the Project Scientist Core Facility is to provide purified proteins, both recombinant and natural, to the Biotechnology Science Team Project Scientists and the NRA-Structural Biology Test Investigators. Having a core facility for this purpose obviates the need for each scientist to develop the necessary expertise and equipment for molecular biology, protein expression, and protein purification. Because of this, they are able to focus their energies as well as their funding on the crystallization and structure determination of their target proteins.

Fiber-Optic Strain-Gage Tank Level Measurement System for Cryogenic Propellants

NASA Technical Reports Server (NTRS)

Figueroa, Fernando; Mitchell, Mark; Langford, Lester

2004-01-01

Measurement of tank level, particularly for cryogenic propellants, has proven to be a difficult problem. Current methods based on differential pressure, capacitance sensors, temperature sensors, etc.; do not provide sufficiently accurate or robust measurements, especially at run time. These methods are designed to measure tank-level, but when the fluids are in supercritical state, the liquid-gas interface disappears. Furthermore, there is a need for a non-intrusive measurement system; that is, the sensors should not require tank modifications and/or disturb the fluids. This paper describes a simple, but effective method to determine propellant mass by measuring very small deformations of the structure supporting the tank. Results of a laboratory study to validate the method, and experimental data from a deployed system are presented. A comparison with an existing differential pressure sensor shows that the strain gage system provides a much better quality signal across all regimes during an engine test. Experimental results also show that the use of fiber optic strain gages (FOSG) over classic foil strain gages extends the operation time (before the system becomes uncalibrated), and increases accuracy. Finally, a procedure is defined whereby measurements from the FOSG mounted on the tank supporting structure are compensated using measurements of a FOSG mounted on a reference plate and temperature measurements of the structure. Results describing the performance of a deployed system that measures tank level during propulsion tests are included.

Low temperature coefficient of resistance and high gage factor in beryllium-doped silicon

NASA Technical Reports Server (NTRS)

Robertson, J. B.; Littlejohn, M. A.

1974-01-01

The gage factor and resistivity of p-type silicon doped with beryllium was studied as a function of temperature, crystal orientation, and beryllium doping concentration. It was shown that the temperature coefficient of resistance can be varied and reduced to zero near room temperature by varying the beryllium doping level. Similarly, the magnitude of the piezoresistance gage factor for beryllium-doped silicon is slightly larger than for silicon doped with a shallow acceptor impurity such as boron, whereas the temperature coefficient of piezoresistance is about the same for material containing these two dopants. These results are discussed in terms of a model for the piezoresistance of compensated p-type silicon.

Linking Statistically- and Physically-Based Models for Improved Streamflow Simulation in Gaged and Ungaged Areas

NASA Astrophysics Data System (ADS)

Lafontaine, J.; Hay, L.; Archfield, S. A.; Farmer, W. H.; Kiang, J. E.

2014-12-01

The U.S. Geological Survey (USGS) has developed a National Hydrologic Model (NHM) to support coordinated, comprehensive and consistent hydrologic model development, and facilitate the application of hydrologic simulations within the continental US. The portion of the NHM located within the Gulf Coastal Plains and Ozarks Landscape Conservation Cooperative (GCPO LCC) is being used to test the feasibility of improving streamflow simulations in gaged and ungaged watersheds by linking statistically- and physically-based hydrologic models. The GCPO LCC covers part or all of 12 states and 5 sub-geographies, totaling approximately 726,000 km2, and is centered on the lower Mississippi Alluvial Valley. A total of 346 USGS streamgages in the GCPO LCC region were selected to evaluate the performance of this new calibration methodology for the period 1980 to 2013. Initially, the physically-based models are calibrated to measured streamflow data to provide a baseline for comparison. An enhanced calibration procedure then is used to calibrate the physically-based models in the gaged and ungaged areas of the GCPO LCC using statistically-based estimates of streamflow. For this application, the calibration procedure is adjusted to address the limitations of the statistically generated time series to reproduce measured streamflow in gaged basins, primarily by incorporating error and bias estimates. As part of this effort, estimates of uncertainty in the model simulations are also computed for the gaged and ungaged watersheds.

Performance benchmarking of four cell-free protein expression systems.

PubMed

Gagoski, Dejan; Polinkovsky, Mark E; Mureev, Sergey; Kunert, Anne; Johnston, Wayne; Gambin, Yann; Alexandrov, Kirill

2016-02-01

Over the last half century, a range of cell-free protein expression systems based on pro- and eukaryotic organisms have been developed and have found a range of applications, from structural biology to directed protein evolution. While it is generally accepted that significant differences in performance among systems exist, there is a paucity of systematic experimental studies supporting this notion. Here, we took advantage of the species-independent translation initiation sequence to express and characterize 87 N-terminally GFP-tagged human cytosolic proteins of different sizes in E. coli, wheat germ (WGE), HeLa, and Leishmania-based (LTE) cell-free systems. Using a combination of single-molecule fluorescence spectroscopy, SDS-PAGE, and Western blot analysis, we assessed the expression yields, the fraction of full-length translation product, and aggregation propensity for each of these systems. Our results demonstrate that the E. coli system has the highest expression yields. However, we observe that high expression levels are accompanied by production of truncated species-particularly pronounced in the case of proteins larger than 70 kDa. Furthermore, proteins produced in the E. coli system display high aggregation propensity, with only 10% of tested proteins being produced in predominantly monodispersed form. The WGE system was the most productive among eukaryotic systems tested. Finally, HeLa and LTE show comparable protein yields that are considerably lower than the ones achieved in the E. coli and WGE systems. The protein products produced in the HeLa system display slightly higher integrity, whereas the LTE-produced proteins have the lowest aggregation propensity among the systems analyzed. The high quality of HeLa- and LTE-produced proteins enable their analysis without purification and make them suitable for analysis of multi-domain eukaryotic proteins. © 2015 Wiley Periodicals, Inc.

Utilizing Photogrammetry and Strain Gage Measurement to Characterize Pressurization of Inflatable Modules

NASA Technical Reports Server (NTRS)

Mohammed, Anil

2011-01-01

This paper focuses on integrating a large hatch penetration into inflatable modules of various constructions. This paper also compares load predictions with test measurements. The strain was measured by utilizing photogrammetric methods and strain gages mounted to select clevises that interface with the structural webbings. Bench testing showed good correlation between strain data collected from an extensometer and photogrammetric measurements, even when the material transitioned from the low load to high load strain region of the curve. The full-scale torus design module showed mixed results as well in the lower load and high strain regions. After thorough analysis of photogrammetric measurements, strain gage measurements, and predicted load, the photogrammetric measurements seem to be off by a factor of two.

Modeling Protein Expression and Protein Signaling Pathways

PubMed Central

Telesca, Donatello; Müller, Peter; Kornblau, Steven M.; Suchard, Marc A.; Ji, Yuan

2015-01-01

High-throughput functional proteomic technologies provide a way to quantify the expression of proteins of interest. Statistical inference centers on identifying the activation state of proteins and their patterns of molecular interaction formalized as dependence structure. Inference on dependence structure is particularly important when proteins are selected because they are part of a common molecular pathway. In that case, inference on dependence structure reveals properties of the underlying pathway. We propose a probability model that represents molecular interactions at the level of hidden binary latent variables that can be interpreted as indicators for active versus inactive states of the proteins. The proposed approach exploits available expert knowledge about the target pathway to define an informative prior on the hidden conditional dependence structure. An important feature of this prior is that it provides an instrument to explicitly anchor the model space to a set of interactions of interest, favoring a local search approach to model determination. We apply our model to reverse-phase protein array data from a study on acute myeloid leukemia. Our inference identifies relevant subpathways in relation to the unfolding of the biological process under study. PMID:26246646

[Construction and expression of a eukaryotic expression vector containing human CR2-Fc fusion protein].

PubMed

Li, Xinxin; Wu, Zhihao; Zhang, Chuanfu; Jia, Leili; Song, Hongbin; Xu, Yuanyong

2014-01-01

To construct a eukaryotic expression vector containing human complement receptor 2 (CR2)-Fc and express the CR2-Fc fusion protein in Chinese hamster ovary (CHO) cells. The extracellular domain of human CR2 and IgG1 Fc were respectively amplified, ligated and inserted into the eukaryotic expression vector PCI-neo. After verified by restriction enzyme digestion and sequencing, the recombinant plasmid was transfected into CHO K1 cells. The ones with stable expression of the fusion protein were obtained by means of G418 selection. The expression of the CR2-Fc fusion protein was detected and confirmed by SDS-PAGE and Western blotting. Restriction enzyme digestion and sequencing demonstrated that the recombinant plasmid was valid. SDS-PAGE showed that relative molecular mass (Mr;) of the purified product was consistent with the expected value. Western blotting further proved the single band at the same position. We constructed the eukaryotic expression vector of CR2-Fc/PCI-neo successfully. The obtained fusion protein was active and can be used for the further study of the role in HIV control.

High-Resolution Rainfall From Radar Reflectivity and Terrestrial Rain Gages for use in Estimating Debris-Flow Susceptibility in the Day Fire, California

NASA Astrophysics Data System (ADS)

Hanshaw, M. N.; Schmidt, K. M.; Jorgensen, D. P.; Stock, J. D.

2007-12-01

Constraining the distribution of rainfall is essential to evaluating the post-fire mass-wasting response of steep soil-mantled landscapes. As part of a pilot early-warning project for flash floods and debris flows, NOAA deployed a portable truck-mounted Shared Mobile Atmospheric Research and Teaching Radar (SMART-R) to the 2006 Day fire in the Transverse Ranges of Southern California. In conjunction with a dense array of ground- based instruments, including 8 tipping-bucket rain gages located within an area of 170 km2, this C-band mobile Doppler radar provided 200-m grid cell estimates of precipitation data at fine temporal and spatial scales in burned steeplands at risk from hazardous flash floods and debris flows. To assess the utility of using this data in process models for flood and debris flow initiation, we converted grids of radar reflectivity to hourly time-steps of precipitation using an empirical relationship for convective storms, sampling the radar data at the locations of each rain gage as determined by GPS. The SMART-R was located 14 km from the farthest rain gage, but <10 km away from our intensive research area, where 5 gages are located within <1-2 km of each other. Analyses of the nine storms imaged by radar throughout the 2006/2007 winter produced similar cumulative rainfall totals between the gages and their SMART-R grid location over the entire season which correlate well on the high side, with gages recording the most precipitation agreeing to within 11% of the SMART-R. In contrast, on the low rainfall side, totals between the two recording systems are more variable, with a 62% variance between the minimums. In addition, at the scale of individual storms, a correlation between ground-based rainfall measurements and radar-based rainfall estimates is less evident, with storm totals between the gages and the SMART-R varying between 7 and 88%, a possible result of these being relatively small, fast-moving storms in an unusually dry winter. The

A Universal Threshold for the Assessment of Load and Output Residuals of Strain-Gage Balance Data

NASA Technical Reports Server (NTRS)

Ulbrich, N.; Volden, T.

2017-01-01

A new universal residual threshold for the detection of load and gage output residual outliers of wind tunnel strain{gage balance data was developed. The threshold works with both the Iterative and Non{Iterative Methods that are used in the aerospace testing community to analyze and process balance data. It also supports all known load and gage output formats that are traditionally used to describe balance data. The threshold's definition is based on an empirical electrical constant. First, the constant is used to construct a threshold for the assessment of gage output residuals. Then, the related threshold for the assessment of load residuals is obtained by multiplying the empirical electrical constant with the sum of the absolute values of all first partial derivatives of a given load component. The empirical constant equals 2.5 microV/V for the assessment of balance calibration or check load data residuals. A value of 0.5 microV/V is recommended for the evaluation of repeat point residuals because, by design, the calculation of these residuals removes errors that are associated with the regression analysis of the data itself. Data from a calibration of a six-component force balance is used to illustrate the application of the new threshold definitions to real{world balance calibration data.

AR-v7 protein expression is regulated by protein kinase and phosphatase

PubMed Central

Li, Yinan; Xie, Ning; Gleave, Martin E.; Rennie, Paul S.; Dong, Xuesen

2015-01-01

Failure of androgen-targeted therapy and progression of castration-resistant prostate cancer (CRPC) are often attributed to sustained expression of the androgen receptor (AR) and its major splice variant, AR-v7. Although the new generation of anti-androgens such as enzalutamide effectively inhibits AR activity, accumulating pre-clinical and clinical evidence indicates that AR-v7 remains constitutively active in driving CRPC progression. However, molecular mechanisms which control AR-v7 protein expression remain unclear. We apply multiple prostate cancer cell models to demonstrate that enzalutamide induces differential activation of protein phosphatase-1 (PP-1) and Akt kinase depending on the gene context of cancer cells. The balance between PP-1 and Akt activation governs AR phosphorylation status and activation of the Mdm2 ubiquitin ligase. Mdm2 recognizes phosphorylated serine 213 of AR-v7, and induces AR-v7 ubiquitination and protein degradation. These findings highlight the decisive roles of PP-1 and Akt for AR-v7 protein expression and activities when AR is functionally blocked. PMID:26378044

SHuffle, a novel Escherichia coli protein expression strain capable of correctly folding disulfide bonded proteins in its cytoplasm

PubMed Central

2012-01-01

Background Production of correctly disulfide bonded proteins to high yields remains a challenge. Recombinant protein expression in Escherichia coli is the popular choice, especially within the research community. While there is an ever growing demand for new expression strains, few strains are dedicated to post-translational modifications, such as disulfide bond formation. Thus, new protein expression strains must be engineered and the parameters involved in producing disulfide bonded proteins must be understood. Results We have engineered a new E. coli protein expression strain named SHuffle, dedicated to producing correctly disulfide bonded active proteins to high yields within its cytoplasm. This strain is based on the trxB gor suppressor strain SMG96 where its cytoplasmic reductive pathways have been diminished, allowing for the formation of disulfide bonds in the cytoplasm. We have further engineered a major improvement by integrating into its chromosome a signal sequenceless disulfide bond isomerase, DsbC. We probed the redox state of DsbC in the oxidizing cytoplasm and evaluated its role in assisting the formation of correctly folded multi-disulfide bonded proteins. We optimized protein expression conditions, varying temperature, induction conditions, strain background and the co-expression of various helper proteins. We found that temperature has the biggest impact on improving yields and that the E. coli B strain background of this strain was superior to the K12 version. We also discovered that auto-expression of substrate target proteins using this strain resulted in higher yields of active pure protein. Finally, we found that co-expression of mutant thioredoxins and PDI homologs improved yields of various substrate proteins. Conclusions This work is the first extensive characterization of the trxB gor suppressor strain. The results presented should help researchers design the appropriate protein expression conditions using SHuffle strains. PMID:22569138

Activation of p44/42 in Human Natural Killer Cells Decreases Cell-surface Protein Expression: Relationship to Tributyltin-induced alterations of protein expression

PubMed Central

Dudimah, Fred D.; Abraha, Abraham; Wang, Xiaofei; Whalen, Margaret M.

2010-01-01

Tributyltin (TBT) activates the mitogen activated protein kinase (MAPK), p44/42 in human natural killer (NK) cells. TBT also reduces NK cytotoxic function and decreases the expression of several NK-cell proteins. To understand the role that p44/42 activation plays in TBT-induced loss of NK cell function, we have investigated how selective activation of p44/42 by phorbol 12-myristate 13-acetate (PMA) affects NK cells. Previously we showed that PMA caused losses of lytic function similar to those seen with TBT exposures. Here we examined activation of p44/42 in the regulation of NK-cell protein expression and how this regulation may explain the protein expression changes seen with TBT exposures. NK cells exposed to PMA were examined for levels of cell-surface proteins, granzyme mRNA, and perforin mRNA expression. The expression of CD11a, CD16, CD18, and CD56 were reduced, perforin mRNA levels were unchanged and granzyme mRNA levels were increased. To verify that activation of p44/42 was responsible for the alterations seen in CD11a, CD16, CD18, and CD56 with PMA, NK cells were treated with the p44/42 pathway inhibitor (PD98059) prior to PMA exposures. In the presence of PD98059, PMA caused no decreases in the expression of the cell-surface proteins. Results of these studies indicate that the activation of p44/42 may lead to the loss of NK cell cytotoxic function by decreasing the expression of CD11a, CD16, CD18, and CD56. Further, activation of p44/42 appears to be at least in part responsible for the TBT-induced decreases in expression of CD16, CD18, and CD56. PMID:20883105

Protein disorder is positively correlated with gene expression in E. coli

PubMed Central

Paliy, Oleg; Gargac, Shawn M.; Cheng, Yugong; Uversky, Vladimir N.; Dunker, A. Keith

2009-01-01

We considered on a global scale the relationship between the predicted fraction of protein disorder and RNA and protein expression in E. coli. Fraction of protein disorder correlated positively with both measured RNA expression levels of E. coli genes in three different growth media and with predicted abundance levels of E. coli proteins. Though weak, the correlation was highly significant. Correlation of protein disorder with RNA expression did not depend on the growth rate of E. coli cultures and was not caused by a small subset of genes showing exceptionally high concordance in their disorder and expression levels. Global analysis was complemented by detailed consideration of several groups of proteins. PMID:18465893

Flood of April 2007 and Flood-Frequency Estimates at Streamflow-Gaging Stations in Western Connecticut

USGS Publications Warehouse

Ahearn, Elizabeth A.

2009-01-01

A spring nor'easter affected the East Coast of the United States from April 15 to 18, 2007. In Connecticut, rainfall varied from 3 inches to more than 7 inches. The combined effects of heavy rainfall over a short duration, high winds, and high tides led to widespread flooding, storm damage, power outages, evacuations, and disruptions to traffic and commerce. The storm caused at least 18 fatalities (none in Connecticut). A Presidential Disaster Declaration was issued on May 11, 2007, for two counties in western Connecticut - Fairfield and Litchfield. This report documents hydrologic and meteorologic aspects of the April 2007 flood and includes estimates of the magnitude of the peak discharges and peak stages during the flood at 28 streamflow-gaging stations in western Connecticut. These data were used to perform flood-frequency analyses. Flood-frequency estimates provided in this report are expressed in terms of exceedance probabilities (the probability of a flood reaching or exceeding a particular magnitude in any year). Flood-frequency estimates for the 0.50, 0.20, 0.10, 0.04, 0.02, 0.01, and 0.002 exceedance probabilities (also expressed as 50-, 20-, 10-, 4-, 2-, 1-, and 0.2- percent exceedance probability, respectively) were computed for 24 of the 28 streamflow-gaging stations. Exceedance probabilities can further be expressed in terms of recurrence intervals (2-, 5-, 10-, 25-, 50-, 100-, and 500-year recurrence interval, respectively). Flood-frequency estimates computed in this study were compared to the flood-frequency estimates used to derive the water-surface profiles in previously published Federal Emergency Management Agency (FEMA) Flood Insurance Studies. The estimates in this report update and supersede previously published flood-frequency estimates for streamflowgaging stations in Connecticut by incorporating additional years of annual peak discharges, including the peaks for the April 2007 flood. In the southwest coastal region of Connecticut, the

46 CFR 32.20-20 - Liquid level gaging-T/ALL.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 46 Shipping 1 2011-10-01 2011-10-01 false Liquid level gaging-T/ALL. 32.20-20 Section 32.20-20 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY TANK VESSELS SPECIAL EQUIPMENT, MACHINERY, AND HULL REQUIREMENTS Equipment Installations § 32.20-20 Liquid level gaging—T/ALL. On tankships, the construction or...

46 CFR 32.20-20 - Liquid level gaging-T/ALL.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 46 Shipping 1 2010-10-01 2010-10-01 false Liquid level gaging-T/ALL. 32.20-20 Section 32.20-20 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY TANK VESSELS SPECIAL EQUIPMENT, MACHINERY, AND HULL REQUIREMENTS Equipment Installations § 32.20-20 Liquid level gaging—T/ALL. On tankships, the construction or...

[Tripartite-motif protein 25 and pyruvate kinase M2 protein expression in non-small cell lung cancer].

PubMed

Jing, Huai-Zhi; Qiu, Feng; Chen, Shi-Zhi; Su, Lin; Qu, Can

2015-03-01

To investigate the expression of tripartite-motif protein 25 (TRIM25) and pyruvate kinase M2 (PKM2) protein in non-small cell lung cancer (NSCLC) and explore their role in the occurrence and progression of NSCLC. The expressions of TRIM25 and PKM2 protein were detected in 60 NSCLC specimens and 20 adjacent normal lung tissue (>5 cm from the lesions) with immunofluorescence histochemical method and in 10 fresh specimens of NSCLC with Western blotting. The results were analyzed in relation with the clinicopathological features of the patients. The positivity rates of TRIM25 expression was 45% in the 60 lung carcinoma specimens, significantly higher than that in the 20 normal lung tissues (10%, P=0.005). TRIM25 protein was expressed in 28.6% of lung adenocarcinoma tissues and in 59.4% of squamous carcinoma tissues (P=0.017). TRIM25 protein expression was positively correlated with the TNM stages and lymph node metastasis of NSCLC (P<0.05). The expressions of PKM2 protein in 60 cases of lung carcinoma was 73.3%,while in 20 cases of normal lung tissues the expressions was 30%(P=0.001). The positivity rates of PKM2 expression differed significantly between lung adenocarcinoma and squamous carcinoma (57.1% vs 87.5%, P=0.008). An inverse correlation was noted between TRIM25 and PKM2 expressions (P=0.026). TRIM25 and PKM2 protein may participate in the occurrence and progression of NSCLC, and their expressions are inversely correlated.

Distinct Protein Expression Profiles of Solid-Pseudopapillary Neoplasms of the Pancreas.

PubMed

Park, Minhee; Lim, Jong-Sun; Lee, Hyoung-Joo; Na, Keun; Lee, Min Jung; Kang, Chang Moo; Paik, Young-Ki; Kim, Hoguen

2015-08-07

Solid-pseudopapillary neoplasm (SPN) is an uncommon pancreatic tumor with mutation in CTNNB1 and distinct clinical and pathological features. We compared the proteomic profiles of SPN to mRNA expression. Pooled SPNs and pooled non-neoplastic pancreatic tissues were examined with high-resolution mass spectrometry. We identified 329 (150 up-regulated and 179 down-regulated) differentially expressed proteins in SPN. We identified 191 proteins (58.1% of the 329 dysregulated proteins) with the same expression tendencies in SPN based on mRNA data. Many overexpressed proteins were related to signaling pathways known to be activated in SPNs. We found that several proteins involved in Wnt signaling, including DKK4 and β-catenin, and proteins that bind β-catenin, such as FUS and NONO, were up-regulated in SPNs. Molecules involved in glycolysis, including PKM2, ENO2, and HK1, were overexpressed in accordance to their mRNA levels. In summary, SPN showed (1) distinct protein expression changes that correlated with mRNA expression, (2) overexpression of Wnt signaling proteins and proteins that bind directly to β-catenin, and (3) overexpression of proteins involved in metabolism. These findings may help develop early diagnostic biomarkers and molecular targets.

Robust expression of a bioactive mammalian protein in chlamydomonas chloroplast

DOEpatents

Mayfield, Stephen P.

2010-03-16

Methods and compositions are disclosed to engineer chloroplast comprising heterologous mammalian genes via a direct replacement of chloroplast Photosystem II (PSII) reaction center protein coding regions to achieve expression of recombinant protein above 5% of total protein. When algae is used, algal expressed protein is produced predominantly as a soluble protein where the functional activity of the peptide is intact. As the host algae is edible, production of biologics in this organism for oral delivery or proteins/peptides, especially gut active proteins, without purification is disclosed.

Robust expression of a bioactive mammalian protein in Chlamydomonas chloroplast

DOEpatents

Mayfield, Stephen P

2015-01-13

Methods and compositions are disclosed to engineer chloroplast comprising heterologous mammalian genes via a direct replacement of chloroplast Photosystem II (PSII) reaction center protein coding regions to achieve expression of recombinant protein above 5% of total protein. When algae is used, algal expressed protein is produced predominantly as a soluble protein where the functional activity of the peptide is intact. As the host algae is edible, production of biologics in this organism for oral delivery of proteins/peptides, especially gut active proteins, without purification is disclosed.

Expression of multiple proteins in transgenic plants

DOEpatents

Vierstra, Richard D.; Walker, Joseph M.

2002-01-01

A method is disclosed for the production of multiple proteins in transgenic plants. A DNA construct for introduction into plants includes a provision to express a fusion protein of two proteins of interest joined by a linking domain including plant ubiquitin. When the fusion protein is produced in the cells of a transgenic plant transformed with the DNA construction, native enzymes present in plant cells cleave the fusion protein to release both proteins of interest into the cells of the transgenic plant. Since the proteins are produced from the same fusion protein, the initial quantities of the proteins in the cells of the plant are approximately equal.

Cancer testis antigens and NY-BR-1 expression in primary breast cancer: prognostic and therapeutic implications.

PubMed

Balafoutas, Dimitrios; zur Hausen, Axel; Mayer, Sebastian; Hirschfeld, Marc; Jaeger, Markus; Denschlag, Dominik; Gitsch, Gerald; Jungbluth, Achim; Stickeler, Elmar

2013-06-03

Cancer-testis antigens (CTA) comprise a family of proteins, which are physiologically expressed in adult human tissues solely in testicular germ cells and occasionally placenta. However, CTA expression has been reported in various malignancies. CTAs have been identified by their ability to elicit autologous cellular and or serological immune responses, and are considered potential targets for cancer immunotherapy. The breast differentiation antigen NY-BR-1, expressed specifically in normal and malignant breast tissue, has also immunogenic properties. Here we evaluated the expression patterns of CTAs and NY-BR-1 in breast cancer in correlation to clinico-pathological parameters in order to determine their possible impact as prognostic factors. The reactivity pattern of various mAbs (6C1, MA454, M3H67, 57B, E978, GAGE #26 and NY-BR-1 #5) were assessed by immunohistochemistry in a tissue micro array series of 210 randomly selected primary invasive breast cancers in order to study the diversity of different CTAs (e.g. MAGE-A, NY-ESO-1, GAGE) and NY-BR-1. These expression data were correlated to clinico-pathological parameters and outcome data including disease-free and overall survival. Expression of at least one CTA was detectable in the cytoplasm of tumor cells in 37.2% of the cases. NY-BR-1 expression was found in 46.6% of tumors, respectively. Overall, CTA expression seemed to be linked to adverse prognosis and M3H67 immunoreactivity specifically was significantly correlated to shorter overall and disease-free survival (p=0.000 and 0.024, respectively). Our findings suggest that M3H67 immunoreactivity could serve as potential prognostic marker in primary breast cancer patients. The exclusive expression of CTAs in tumor tissues as well as the frequent expression of NY-BR-1 could define new targets for specific breast cancer therapies.

Cost-effectiveness of the US Geological Survey stream-gaging program in Arkansas

USGS Publications Warehouse

Darling, M.E.; Lamb, T.E.

1984-01-01

This report documents the results of the cost-effectiveness of the stream-gaging program in Arkansas. Data uses and funding sources were identified for the daily-discharge stations. All daily-discharge stations were found to be in one or more data use categories, and none were candidates for alternate methods which would result in discontinuation or conversion to a partial record station. The cost for operation of daily-discharge stations and routing costs to partial record stations, crest gages, pollution control stations as well as seven recording ground-water stations was evaluated in the Kalman-Filtering Cost-Effective Resource allocation (K-CERA) analysis. This operation under current practices requires a budget of $292,150. The average standard error of estimate of streamflow record for the Arkansas District was analyzed at 33 percent.

Design of Gages for Direct Skin Friction Measurements in Complex Turbulent Flows with Shock Impingement Compensation

DTIC Science & Technology

2007-06-07

100 kW/m2 for 0.1 s. Along with the material change, an oil leak problem required a geometric change. Initially, we considered TIG welding or...shear and moment, is addressed through the design, development, and testing of the CF1 and CF2 gages. Chapter 3 presents the evolutionary process ...a shock. Chapter 4 examines the performance of each gage to the nominal load conditions. Through this process , objective 2 is met. The best

Methods and constructs for expression of foreign proteins in photosynthetic organisms

DOEpatents

Laible, Philip D.; Hanson, Deborah K.

2002-01-01

A method for expressing and purifying foreign proteins in photosynthetic organisms comprising the simultaneous expression of both the heterologous protein and a means for compartmentalizing or sequestering of the protein.

The operation and maintenance of a crest-stage gaging station

USGS Publications Warehouse

Friday, John

1965-01-01

Rigid datum controls must be maintained at the gage site throughout the period of record. Physical changes of the site resulting from flood flows or manmade alterations must be evaluated. If a drainage structure such as a culvert is part of the site features, free-flow conditions must be maintained or obstructions carefully documented.

Similar protein expression profiles of ovarian and endometrial high-grade serous carcinomas.

PubMed

Hiramatsu, Kosuke; Yoshino, Kiyoshi; Serada, Satoshi; Yoshihara, Kosuke; Hori, Yumiko; Fujimoto, Minoru; Matsuzaki, Shinya; Egawa-Takata, Tomomi; Kobayashi, Eiji; Ueda, Yutaka; Morii, Eiichi; Enomoto, Takayuki; Naka, Tetsuji; Kimura, Tadashi

2016-03-01

Ovarian and endometrial high-grade serous carcinomas (HGSCs) have similar clinical and pathological characteristics; however, exhaustive protein expression profiling of these cancers has yet to be reported. We performed protein expression profiling on 14 cases of HGSCs (7 ovarian and 7 endometrial) and 18 endometrioid carcinomas (9 ovarian and 9 endometrial) using iTRAQ-based exhaustive and quantitative protein analysis. We identified 828 tumour-expressed proteins and evaluated the statistical similarity of protein expression profiles between ovarian and endometrial HGSCs using unsupervised hierarchical cluster analysis (P<0.01). Using 45 statistically highly expressed proteins in HGSCs, protein ontology analysis detected two enriched terms and proteins composing each term: IMP2 and MCM2. Immunohistochemical analyses confirmed the higher expression of IMP2 and MCM2 in ovarian and endometrial HGSCs as well as in tubal and peritoneal HGSCs than in endometrioid carcinomas (P<0.01). The knockdown of either IMP2 or MCM2 by siRNA interference significantly decreased the proliferation rate of ovarian HGSC cell line (P<0.01). We demonstrated the statistical similarity of the protein expression profiles of ovarian and endometrial HGSC beyond the organs. We suggest that increased IMP2 and MCM2 expression may underlie some of the rapid HGSC growth observed clinically.

The protein expression landscape of the Arabidopsis root

PubMed Central

Petricka, Jalean J.; Schauer, Monica A.; Megraw, Molly; Breakfield, Natalie W.; Thompson, J. Will; Georgiev, Stoyan; Soderblom, Erik J.; Ohler, Uwe; Moseley, Martin Arthur; Grossniklaus, Ueli; Benfey, Philip N.

2012-01-01

Because proteins are the major functional components of cells, knowledge of their cellular localization is crucial to gaining an understanding of the biology of multicellular organisms. We have generated a protein expression map of the Arabidopsis root providing the identity and cell type-specific localization of nearly 2,000 proteins. Grouping proteins into functional categories revealed unique cellular functions and identified cell type-specific biomarkers. Cellular colocalization provided support for numerous protein–protein interactions. With a binary comparison, we found that RNA and protein expression profiles are weakly correlated. We then performed peak integration at cell type-specific resolution and found an improved correlation with transcriptome data using continuous values. We performed GeLC-MS/MS (in-gel tryptic digestion followed by liquid chromatography-tandem mass spectrometry) proteomic experiments on mutants with ectopic and no root hairs, providing complementary proteomic data. Finally, among our root hair-specific proteins we identified two unique regulators of root hair development. PMID:22447775

Trichohyalin-like 1 protein, a member of fused S100 proteins, is expressed in normal and pathologic human skin

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yamakoshi, Takako; Makino, Teruhiko, E-mail: tmakino@med.u-toyama.ac.jp; Ur Rehman, Mati

2013-03-01

Highlights: ► Trichohyalin-like 1 protein is a member of the fused-type S100 protein gene family. ► Specific antibodies against the C-terminus of the TCHHL1 protein were generated. ► TCHHL1 proteins were expressed in the basal layer of the normal epidermis. ► TCHHL1 proteins were strongly expressed in tumor nests of BCC and SCC. ► The expression of TCHHL1 proteins increased in epidermis of psoriasis vulgaris. - Abstract: Trichohyalin-like 1 (TCHHL1) protein is a novel member of the fused-type S100 protein gene family. The deduced amino acid sequence of TCHHL1 contains an EF-hand domain in the N-terminus, one trans-membrane domain andmore » a nuclear localization signal. We generated specific antibodies against the C-terminus of the TCHHL1 protein and examined the expression of TCHHL1 proteins in normal and pathological human skin. An immunohistochemical study showed that TCHHL1 proteins were expressed in the basal layer of the normal epidermis. In addition, signals of TCHHL1 proteins were observed around the nuclei of cultured growing keratinocytes. Accordingly, TCHHL1 mRNA has been detected in normal skin and cultured growing keratinocytes. Furthermore, TCHHL1 proteins were strongly expressed in the peripheral areas of tumor nests in basal cell carcinomas and squamous cell carcinomas. A dramatic increase in the number of Ki67 positive cells was observed in TCHHL1-expressing areas. The expression of TCHHL1 proteins also increased in non-cancerous hyperproliferative epidermal tissues such as those of psoriasis vulgaris and lichen planus. These findings highlight the possibility that TCHHL1 proteins are expressed in growing keratinocytes of the epidermis and might be associated with the proliferation of keratinocytes.« less

Transient Expression of Chimeric Fluorescent Reporter Proteins in Pollen Tubes to Study Protein Polar Secretion and Dynamics.

PubMed

Zhong, Guitao; Liu, Ronghe; Zhuang, Menglong; Wang, Hao

2017-01-01

Transient expression of chimeric fluorescent reporter proteins by biolistic bombardment is a quick and useful procedure for studying subcellular protein localization and dynamics in plants. It is especially beneficial in specific plant cells which are not suitable for protoplast-based and Agrobacterium-mediated protein transient expression. Polar protein secretion and vesicular trafficking play essential functions for cell polarization and tip growth. The growing pollen tube is regarded as an ideal model plant cell system to study the machinery and regulation of polar protein trafficking and targeting. A large amount of newly synthesized proteins are packed and polarly transported to the apical region to support the rapid and highly polarized tip growth. Here, we described a detailed step-by-step protocol for the transient expression of chimeric fluorescent reporter proteins in growing Arabidopsis and tobacco pollen tubes to study polar transportation logistics and mechanisms. In addition, we have optimized the Arabidopsis and tobacco in vitro pollen germination medium and the conditions to maximize the efficiency of protein expression. As a proof of concept, we have used this protocol to express actin microfilament and late endosomal fluorescent markers in Arabidopsis and tobacco pollen tubes.

Impact of Chronic Alcohol Ingestion on Cardiac Muscle Protein Expression

PubMed Central

Fogle, Rachel L.; Lynch, Christopher J.; Palopoli, Mary; Deiter, Gina; Stanley, Bruce A.; Vary, Thomas C.

2014-01-01

Background Chronic alcohol abuse contributes not only to an increased risk of health-related complications, but also to a premature mortality in adults. Myocardial dysfunction, including the development of a syndrome referred to as alcoholic cardiomyopathy, appears to be a major contributing factor. One mechanism to account for the pathogenesis of alcoholic cardiomyopathy involves alterations in protein expression secondary to an inhibition of protein synthesis. However, the full extent to which myocardial proteins are affected by chronic alcohol consumption remains unresolved. Methods The purpose of this study was to examine the effect of chronic alcohol consumption on the expression of cardiac proteins. Male rats were maintained for 16 weeks on a 40% ethanol-containing diet in which alcohol was provided both in drinking water and agar blocks. Control animals were pair-fed to consume the same caloric intake. Heart homogenates from control- and ethanol-fed rats were labeled with the cleavable isotope coded affinity tags (ICAT™). Following the reaction with the ICAT™ reagent, we applied one-dimensional gel electrophoresis with in-gel trypsin digestion of proteins and subsequent MALDI-TOF-TOF mass spectrometric techniques for identification of peptides. Differences in the expression of cardiac proteins from control- and ethanol-fed rats were determined by mass spectrometry approaches. Results Initial proteomic analysis identified and quantified hundreds of cardiac proteins. Major decreases in the expression of specific myocardial proteins were observed. Proteins were grouped depending on their contribution to multiple activities of cardiac function and metabolism, including mitochondrial-, glycolytic-, myofibrillar-, membrane-associated, and plasma proteins. Another group contained identified proteins that could not be properly categorized under the aforementioned classification system. Conclusions Based on the changes in proteins, we speculate modulation of

High-level expression of soluble recombinant proteins in Escherichia coli using an HE-maltotriose-binding protein fusion tag.

PubMed

Han, Yingqian; Guo, Wanying; Su, Bingqian; Guo, Yujie; Wang, Jiang; Chu, Beibei; Yang, Guoyu

2018-02-01

Recombinant proteins are commonly expressed in prokaryotic expression systems for large-scale production. The use of genetically engineered affinity and solubility enhancing fusion proteins has increased greatly in recent years, and there now exists a considerable repertoire of these that can be used to enhance the expression, stability, solubility, folding, and purification of their fusion partner. Here, a modified histidine tag (HE) used as an affinity tag was employed together with a truncated maltotriose-binding protein (MBP; consisting of residues 59-433) from Pyrococcus furiosus as a solubility enhancing tag accompanying a tobacco etch virus protease-recognition site for protein expression and purification in Escherichia coli. Various proteins tagged at the N-terminus with HE-MBP(Pyr) were expressed in E. coli BL21(DE3) cells to determine expression and solubility relative to those tagged with His6-MBP or His6-MBP(Pyr). Furthermore, four HE-MBP(Pyr)-fused proteins were purified by immobilized metal affinity chromatography to assess the affinity of HE with immobilized Ni 2+ . Our results showed that HE-MBP(Pyr) represents an attractive fusion protein allowing high levels of soluble expression and purification of recombinant protein in E. coli. Copyright © 2017 Elsevier Inc. All rights reserved.

Assessing the utility of gene co-expression stability in combination with correlation in the analysis of protein-protein interaction networks

PubMed Central

2011-01-01

Background Gene co-expression, in the form of a correlation coefficient, has been valuable in the analysis, classification and prediction of protein-protein interactions. However, it is susceptible to bias from a few samples having a large effect on the correlation coefficient. Gene co-expression stability is a means of quantifying this bias, with high stability indicating robust, unbiased co-expression correlation coefficients. We assess the utility of gene co-expression stability as an additional measure to support the co-expression correlation in the analysis of protein-protein interaction networks. Results We studied the patterns of co-expression correlation and stability in interacting proteins with respect to their interaction promiscuity, levels of intrinsic disorder, and essentiality or disease-relatedness. Co-expression stability, along with co-expression correlation, acts as a better classifier of hub proteins in interaction networks, than co-expression correlation alone, enabling the identification of a class of hubs that are functionally distinct from the widely accepted transient (date) and obligate (party) hubs. Proteins with high levels of intrinsic disorder have low co-expression correlation and high stability with their interaction partners suggesting their involvement in transient interactions, except for a small group that have high co-expression correlation and are typically subunits of stable complexes. Similar behavior was seen for disease-related and essential genes. Interacting proteins that are both disordered have higher co-expression stability than ordered protein pairs. Using co-expression correlation and stability, we found that transient interactions are more likely to occur between an ordered and a disordered protein while obligate interactions primarily occur between proteins that are either both ordered, or disordered. Conclusions We observe that co-expression stability shows distinct patterns in structurally and functionally

Frame-Insensitive Expression Cloning of Fluorescent Protein from Scolionema suvaense.

PubMed

Horiuchi, Yuki; Laskaratou, Danai; Sliwa, Michel; Ruckebusch, Cyril; Hatori, Kuniyuki; Mizuno, Hideaki; Hotta, Jun-Ichi

2018-01-26

Expression cloning from cDNA is an important technique for acquiring genes encoding novel fluorescent proteins. However, the probability of in-frame cDNA insertion following the first start codon of the vector is normally only 1/3, which is a cause of low cloning efficiency. To overcome this issue, we developed a new expression plasmid vector, pRSET-TriEX, in which transcriptional slippage was induced by introducing a DNA sequence of (dT) 14 next to the first start codon of pRSET. The effectiveness of frame-insensitive cloning was validated by inserting the gene encoding eGFP with all three possible frames to the vector. After transformation with one of these plasmids, E. coli cells expressed eGFP with no significant difference in the expression level. The pRSET-TriEX vector was then used for expression cloning of a novel fluorescent protein from Scolionema suvaense . We screened 3658 E. coli colonies transformed with pRSET-TriEX containing Scolionema suvaense cDNA, and found one colony expressing a novel green fluorescent protein, ScSuFP. The highest score in protein sequence similarity was 42% with the chain c of multi-domain green fluorescent protein like protein "ember" from Anthoathecata sp. Variations in the N- and/or C-terminal sequence of ScSuFP compared to other fluorescent proteins indicate that the expression cloning, rather than the sequence similarity-based methods, was crucial for acquiring the gene encoding ScSuFP. The absorption maximum was at 498 nm, with an extinction efficiency of 1.17 × 10⁵ M -1 ·cm -1 . The emission maximum was at 511 nm and the fluorescence quantum yield was determined to be 0.6. Pseudo-native gel electrophoresis showed that the protein forms obligatory homodimers.

A Dual-Range Strain Gage Weighing Transducer Employing Automatic Switching

Treesearch

Rodger A. Arola

1968-01-01

Describes a dual-range strain gage transducer which has proven to be an excellent weight-sensing device for weighing trees and tree-length logs; discusses basic principals of the design and operation; and shows that a single transducer having two sensitivity ranges with automatic internal switching can sense weight with good repeatability and that one calibration curve...

The Shuttle Orbital Maneuvering System P-V-T Propellant Quantity Gaging Accuracy and Leak Detection Allowance for Four Instrumentation Conditions

NASA Technical Reports Server (NTRS)

Duhon, D. D.

1975-01-01

The shuttle orbital maneuvering system (OMS) pressure-volume-temperature (P-V-T) propellant gaging module computes the quantity of usable OMS propellant remaining based on the real gas P-V-T relationship for the propellant tank pressurant, helium. The OMS P-V-T propellant quantity gaging error was determined for four sets of instrumentation configurations and accuracies with the propellant tank operating in the normal constant pressure mode and in the blowdown mode. The instrumentation inaccuracy allowance for propellant leak detection was also computed for these same four sets of instrumentation. These gaging errors and leak detection allowances are presented in tables designed to permit a direct comparison of the effectiveness of the four instrumentation sets. The results show the magnitudes of the improvements in propellant quantity gaging accuracies and propellant leak detection allowances which can be achieved by employing more accurate pressure and temperature instrumentation.

Enteral delivery of proteins enhances the expression of proteins involved in the cytoskeleton and protein biosynthesis in human duodenal mucosa.

PubMed

Goichon, Alexis; Bertrand, Julien; Chan, Philippe; Lecleire, Stéphane; Coquard, Aude; Cailleux, Anne-Françoise; Vaudry, David; Déchelotte, Pierre; Coëffier, Moïse

2015-08-01

Amino acids are well known to be key effectors of gut protein turnover. We recently reported that enteral delivery of proteins markedly stimulated global duodenal protein synthesis in carbohydrate-fed healthy humans, but specifically affected proteins remain unknown. We aimed to assess the influence of an enteral protein supply on the duodenal mucosal proteome in carbohydrate-fed humans. Six healthy volunteers received for 5 h, on 2 occasions and in random order, either an enteral infusion of maltodextrins alone (0.25 g · kg⁻¹ · h⁻¹) mimicking the fed state or maltodextrins with a protein powder (0.14 g proteins · kg⁻¹ · h⁻¹). Endoscopic duodenal biopsy specimens were then collected and frozen until analysis. A 2-dimensional polyacrylamide gel electrophoresis-based comparative proteomics analysis was then performed, and differentially expressed proteins (at least ±1.5-fold change; Student's t test, P < 0.05) were identified by mass spectrometry. Protein expression changes were confirmed by Western blot analysis. Thirty-two protein spots were differentially expressed after protein delivery compared with maltodextrins alone: 28 and 4 spots were up- or downregulated, respectively. Among the 22 identified proteins, 11 upregulated proteins were involved either in the cytoskeleton (ezrin, moesin, plastin 1, lamin B1, vimentin, and β-actin) or in protein biosynthesis (glutamyl-prolyl-transfer RNA synthetase, glutaminyl-transfer RNA synthetase, elongation factor 2, elongation factor 1δ, and eukaryotic translation and initiation factor 3 subunit f). Enteral delivery of proteins altered the duodenal mucosal proteome and mainly stimulated the expression of proteins involved in cytoskeleton and protein biosynthesis. These results suggest that protein supply may affect intestinal morphology by stimulating actin cytoskeleton remodeling. © 2015 American Society for Nutrition.

Phytomonas: A non-pathogenic trypanosomatid model for functional expression of proteins.

PubMed

Miranda, Mariana R; Sayé, Melisa; Reigada, Chantal; Carrillo, Carolina; Pereira, Claudio A

2015-10-01

Phytomonas are protozoan parasites from the Trypanosomatidae family which infect a wide variety of plants. Herein, Phytomonas Jma was tested as a model for functional expression of heterologous proteins. Green fluorescent protein expression was evaluated in Phytomonas and compared with Trypanosoma cruzi, the etiological agent of Chagas' disease. Phytomonas was able to express GFP at levels similar to T. cruzi although the transgenic selection time was higher. It was possible to establish an efficient transfection and selection protocol for protein expression. These results demonstrate that Phytomonas can be a good model for functional expression of proteins from other trypanosomatids, presenting the advantage of being completely safe for humans. Copyright © 2015 Elsevier Inc. All rights reserved.

Cryogenic strain gage techniques used in force balance design for the National Transonic Facility

NASA Technical Reports Server (NTRS)

Ferris, A. T.

1986-01-01

A force balance is a strain gage transducer used in wind tunnels to measure the forces and moments on aerodynamic models. Techniques have been established for temperature-compensation of force balances to allow their use over the operating temperature range of a cryogenic wind tunnel (-190C to 60C) without thermal control. This was accomplished by using a patented strain gage matching process to minimize inherent thermal differences, and a thermal compensation procedure to reduce the remaining thermally-induced outputs to acceptable levels. A method of compensating for mechanical movement of the axial force measuring beam caused by thermally-induced stresses under transient temperatures was also included.

The effects of incomplete annealing on the temperature dependence of sheet resistance and gage factor in aluminum and phosphorus implanted silicon on sapphire

NASA Technical Reports Server (NTRS)

Pisciotta, B. P.; Gross, C.

1976-01-01

Partial annealing of damage to the crystal lattice during ion implantation reduces the temperature coefficient of resistivity of ion-implanted silicon, while facilitating controlled doping. Reliance on this method for temperature compensation of the resistivity and strain-gage factor is discussed. Implantation conditions and annealing conditions are detailed. The gage factor and its temperature variation are not drastically affected by crystal damage for some crystal orientations. A model is proposed to account for the effects of electron damage on the temperature dependence of resistivity and on silicon piezoresistance. The results are applicable to the design of silicon-on-sapphire strain gages with high gage factors.

Feasibility of development of a nuclear density gage for determining the density of plastic concrete at a particular stratum : final report.

DOT National Transportation Integrated Search

1981-05-01

Development of a nuclear density gage for determining the degree of consolidation of plastic concrete in selected horizontal layers was determined to be feasible. A commercially available, with some modifications, two-probe density gage (detector in ...

Cost effectiveness of the US Geological Survey stream-gaging program in Alabama

USGS Publications Warehouse

Jeffcoat, H.H.

1987-01-01

A study of the cost effectiveness of the stream gaging program in Alabama identified data uses and funding sources for 72 surface water stations (including dam stations, slope stations, and continuous-velocity stations) operated by the U.S. Geological Survey in Alabama with a budget of $393,600. Of these , 58 gaging stations were used in all phases of the analysis at a funding level of $328,380. For the current policy of operation of the 58-station program, the average standard error of estimation of instantaneous discharge is 29.3%. This overall level of accuracy can be maintained with a budget of $319,800 by optimizing routes and implementing some policy changes. The maximum budget considered in the analysis was $361,200, which gave an average standard error of estimation of 20.6%. The minimum budget considered was $299,360, with an average standard error of estimation of 36.5%. The study indicates that a major source of error in the stream gaging records is lost or missing data that are the result of streamside equipment failure. If perfect equipment were available, the standard error in estimating instantaneous discharge under the current program and budget could be reduced to 18.6%. This can also be interpreted to mean that the streamflow data records have a standard error of this magnitude during times when the equipment is operating properly. (Author 's abstract)

High-throughput Cloning and Expression of Integral Membrane Proteins in Escherichia coli

PubMed Central

Bruni, Renato

2014-01-01

Recently, several structural genomics centers have been established and a remarkable number of three-dimensional structures of soluble proteins have been solved. For membrane proteins, the number of structures solved has been significantly trailing those for their soluble counterparts, not least because over-expression and purification of membrane proteins is a much more arduous process. By using high throughput technologies, a large number of membrane protein targets can be screened simultaneously and a greater number of expression and purification conditions can be employed, leading to a higher probability of successfully determining the structure of membrane proteins. This unit describes the cloning, expression and screening of membrane proteins using high throughput methodologies developed in our laboratory. Basic Protocol 1 deals with the cloning of inserts into expression vectors by ligation-independent cloning. Basic Protocol 2 describes the expression and purification of the target proteins on a miniscale. Lastly, for the targets that express at the miniscale, basic protocols 3 and 4 outline the methods employed for the expression and purification of targets at the midi-scale, as well as a procedure for detergent screening and identification of detergent(s) in which the target protein is stable. PMID:24510647

Expression of hypoxia-inducible 2 (HIG2) protein in uterine cancer.

PubMed

Nishimura, S; Tsuda, H; Nomura, H; Kataoka, F; Chiyoda, T; Tanaka, H; Tanaka, K; Susumu, N; Aoki, D

2011-01-01

For both cervical cancer (UCC) and endometrial cancer (EMC) there are no effective prognostic markers. In this study, we evaluated HIG2 protein expression in 332 uterine cancers (186 UCCs and 146 EMCs) and examined the relationship between HIG2 protein expression and clinical factors, including prognosis. Totally, HIG2 expression was detected in 58% of UCC and 66% of EMC. However, there was no significant relationship between HIG2 expression and age, clinical stage and histology in either UCC or EMC. In addition, HIG2 protein expression was not related to prognosis of UCC or EMC. The positivity rate of HIG2 protein was 56% and 61% in early-stage UCC and EMC, respectively and 67% in non-squamous cell carcinoma of UCC. The positivity rate of HIG2 protein was high even in early-stage UCC and EMC

Condition assessment of reinforced concrete beams using dynamic data measured with distributed long-gage macro-strain sensors

NASA Astrophysics Data System (ADS)

Hong, W.; Wu, Z. S.; Yang, C. Q.; Wan, C. F.; Wu, G.; Zhang, Y. F.

2012-06-01

A new condition assessment strategy of reinforced concrete (RC) beams is proposed in this paper. This strategy is based on frequency analysis of the dynamic data measured with distributed long-gage macro-stain sensors. After extracting modal macro-strain, the reference-based damage index is theoretically deducted in which the variations of modal flexural rigidity and modal neutral axis height are considered. The reference-free damage index is also presented for comparison. Both finite element simulation and experiment investigations were carried out to verify the proposed method. The manufacturing procedure of long-gage fiber Bragg grating (FBG) sensor chosen in the experiment is firstly presented, followed by an experimental study on the essential sensing properties of the long-gage macro-strain sensors and the results verify the excellent sensing properties, in particular the measurement accuracy and dynamic measuring capacity. Modal analysis results of a concrete beam show that the damage appearing in the beam can be well identified by the damage index while the vibration testing results of a RC beam show that the proposed method can not only capture small crack initiation but its propagation. It can be concluded that distributed long-gage dynamic macro-strain sensing technique has great potential for the condition assessment of RC structures subjected to dynamic loading.

The unique C- and N-terminal sequences of Metallothionein isoform 3 mediate growth inhibition and Vectorial active transport in MCF-7 cells.

PubMed

Voels, Brent; Wang, Liping; Sens, Donald A; Garrett, Scott H; Zhang, Ke; Somji, Seema

2017-05-25

The 3rd isoform of the metallothionein (MT3) gene family has been shown to be overexpressed in most ductal breast cancers. A previous study has shown that the stable transfection of MCF-7 cells with the MT3 gene inhibits cell growth. The goal of the present study was to determine the role of the unique C-terminal and N-terminal sequences of MT3 on phenotypic properties and gene expression profiles of MCF-7 cells. MCF-7 cells were transfected with various metallothionein gene constructs which contain the insertion or the removal of the unique MT3 C- and N-terminal domains. Global gene expression analysis was performed on the MCF-7 cells containing the various constructs and the expression of the unique C- and N- terminal domains of MT3 was correlated to phenotypic properties of the cells. The results of the present study demonstrate that the C-terminal sequence of MT3, in the absence of the N-terminal sequence, induces dome formation in MCF-7 cells, which in cell cultures is the phenotypic manifestation of a cell's ability to perform vectorial active transport. Global gene expression analysis demonstrated that the increased expression of the GAGE gene family correlated with dome formation. Expression of the C-terminal domain induced GAGE gene expression, whereas the N-terminal domain inhibited GAGE gene expression and that the effect of the N-terminal domain inhibition was dominant over the C-terminal domain of MT3. Transfection with the metallothionein 1E gene increased the expression of GAGE genes. In addition, both the C- and the N-terminal sequences of the MT3 gene had growth inhibitory properties, which correlated to an increased expression of the interferon alpha-inducible protein 6. Our study shows that the C-terminal domain of MT3 confers dome formation in MCF-7 cells and the presence of this domain induces expression of the GAGE family of genes. The differential effects of MT3 and metallothionein 1E on the expression of GAGE genes suggests unique roles of

Transient expression of CCL21as recombinant protein in tomato.

PubMed

Beihaghi, Maria; Marashi, Hasan; Bagheri, Abdolreza; Sankian, Mojtaba

2018-03-01

The main goal of this study was to investigate the possibility of expressing recombinant protein of C-C chemokine ligand 21 (CCL21) in Solanum lycopersicum via agroinfiltration. CCL21 is a chemokine can be used for anti-metastatic of cancer cell lines. To examine the expression of CCL21 protein in S. lycopersicum , the construct of ccl21 was synthesized. This construct was cloned into pBI121 and the resulting CCL21 plasmid was agro-infiltrated into S. lycopersicum leaves. Within three days after infiltration, Expression of the foreign gene was confirmed by quantitative Real-time PCR. A recombinant CCL21 protein was immunogenically detected by western blot, dot blot and ELISA assay. And results showed that the foreign gene was expressed in the transformed leaves in high level. Also scratch assay was used to investigate the role of this protein in anti-metastatic function. The results demonstrated anti-metastatic of cancer cells in the presence of this protein.

Alfred P. Gage and the Introductory Physics Laboratory

NASA Astrophysics Data System (ADS)

Greenslade, Thomas B.

2016-03-01

This article is about a late 19th-century teacher of secondary school physics. I was originally interested in the apparatus that he sold. This led me to the physics books that he wrote, and these took me to his unusual ideas about ways to use laboratory time to introduce students to the phenomena of physics. More than 100 years later educational ideas have now come full circle, and it is time to bring Gage and his texts and ideas to 21st-century physics teachers.

Transient Expression and Cellular Localization of Recombinant Proteins in Cultured Insect Cells.

PubMed

Fabrick, Jeffrey A; Hull, J Joe

2017-04-20

Heterologous protein expression systems are used for the production of recombinant proteins, the interpretation of cellular trafficking/localization, and the determination of the biochemical function of proteins at the sub-organismal level. Although baculovirus expression systems are increasingly used for protein production in numerous biotechnological, pharmaceutical, and industrial applications, nonlytic systems that do not involve viral infection have clear benefits but are often overlooked and underutilized. Here, we describe a method for generating nonlytic expression vectors and transient recombinant protein expression. This protocol allows for the efficient cellular localization of recombinant proteins and can be used to rapidly discern protein trafficking within the cell. We show the expression of four recombinant proteins in a commercially available insect cell line, including two aquaporin proteins from the insect Bemisia tabaci, as well as subcellular marker proteins specific for the cell plasma membrane and for intracellular lysosomes. All recombinant proteins were produced as chimeras with fluorescent protein markers at their carboxyl termini, which allows for the direct detection of the recombinant proteins. The double transfection of cells with plasmids harboring constructs for the genes of interest and a known subcellular marker allows for live cell imaging and improved validation of cellular protein localization.

Use of a protein engineering strategy to overcome limitations in the production of "Difficult to Express" recombinant proteins.

PubMed

Hussain, Hirra; Fisher, David I; Abbott, W Mark; Roth, Robert G; Dickson, Alan J

2017-10-01

Certain recombinant proteins are deemed "difficult to express" in mammalian expression systems requiring significant cell and/or process engineering to abrogate expression bottlenecks. With increasing demand for the production of recombinant proteins in mammalian cells, low protein yields can have significant consequences for industrial processes. To investigate the molecular mechanisms that restrict expression of recombinant proteins, naturally secreted model proteins were analyzed from the tissue inhibitors of metalloproteinase (TIMP) protein family. In particular, TIMP-2 and TIMP-3 were subjected to detailed study. TIMP proteins share significant sequence homology (∼50% identity and ∼70% similarity in amino acid sequence). However, they show marked differences in secretion in mammalian expression systems despite this extensive sequence homology. Using these two proteins as models, this study characterized the molecular mechanisms responsible for poor recombinant protein production. Our results reveal that both TIMP-2 and TIMP-3 are detectable at mRNA and protein level within the cell but only TIMP-2 is secreted effectively into the extracellular medium. Analysis of protein localization and the nature of intracellular protein suggest TIMP-3 is severely limited in its post-translational processing. To overcome this challenge, modification of the TIMP-3 sequence to include a furin protease-cleavable pro-sequence resulted in secretion of the modified TIMP-3 protein, however, incomplete processing was observed. Based on the TIMP-3 data, the protein engineering approach was optimized and successfully applied in combination with cell engineering, the overexpression of furin, to another member of the TIMP protein family (the poorly expressed TIMP-4). Use of the described protein engineering strategy resulted in successful secretion of poorly (TIMP-4) and non-secreted (TIMP-3) targets, and presents a novel strategy to enhance the production of "difficult" recombinant

Refinement of moisture calibration curves for nuclear gage : interim report no. 1.

DOT National Transportation Integrated Search

1972-01-01

This study was initiated to determine the correct moisture calibration curves for different nuclear gages. It was found that the Troxler Model 227 had a linear response between count ratio and moisture content. Also, the two calibration curves for th...

Thymidylate synthase (TS) protein expression as a prognostic factor in advanced colorectal cancer: a comparison with TS mRNA expression.

PubMed

Nakagawa, Tateo; Shimada, Mitsuo; Kurita, Nobuhiro; Iwata, Takashi; Nishioka, Masanori; Yoshikawa, Kozo; Higashijima, Jun; Utsunomiya, Tohru

2012-06-01

The role of intratumoral thymidylate synthase (TS) mRNA or protein expression is still controversial and little has been reported regarding relation of them in colorectal cancer. Forty-six patients with advanced colorectal cancer who underwent surgical resection were included. TS mRNA expression was determined by the Danenberg tumor profile method based on laser-captured micro-dissection of the tumor cells. TS protein expression was evaluated using immunohistochemical staining. TS mRNA expression tended to relate TS protein expression. Statistical significance was not found in overall survival between the TS mRNA high group and low group regardless of performing adjuvant chemotherapy. The overall survival in the TS protein negative group was significantly higher than that in positive group in all and the patients without adjuvant chemotherapy. Multivariate analysis showed TS protein expression was as an independent prognostic factor. TS protein expression tends to be related TS mRNA expression and is an independent prognostic factor in advanced colorectal cancer.

Determination of base-flow characteristics at selected streamflow-gaging stations on the Mad River, Ohio

USGS Publications Warehouse

Koltun, G.F.

1995-01-01

This report describes the results of a study to estimate characteristics of base flow and sustained ground-water discharge at five streamflow-gaging stations on the Mad River in Ohio. The five streamflow-gaging stations are located at Zanesfield, near Urbana, at St. Paris Pike (at Eagle City), near Springfield, and near Dayton. The median of the annual-mean base flows, determined by means of hydrograph separation, ranged from 0.64 (ft3/s)/mi2 (cubic feet per second per square mile) at Zanesfield to 0.74 (ft3/s)/mi2 at St. Paris Pike. The median percentage of annual total streamflow attributed to base flow ranged from 61.8 percent at Zanesfield to 76.1 percent near Urbana. Estimates of an upper limit (or threshold) at which base flows can be considered to be composed predominately of sustained ground-water discharge were made by constructing and analyzing base- flow-duration curves. The sustained ground-water discharges (base flows less than or equal to the estimated sustained ground-water-discharge thresholds) are assumed to originate from ground-water- flow systems that are minimally affected by seasonal climatic changes. The median sustained ground- water discharge ranged from 0.11 (ft3/s)/mi2 at Zanesfield to 0.26 (ft3/s)/mi2 at St. Paris Pike (at Eagle City) and near Springfield. The median sustained ground-water discharge, expressed as a percentage of the median annual-mean base flow, ranged from 17.2 percent at Zanesfield to 38.6 percent near Springfield.

Cloning and expression of Tenebrio molitor antifreeze protein in Escherichia coli.

PubMed

Yue, Chang-Wu; Zhang, Yi-Zheng

2009-03-01

A novel antifreeze protein cDNA was cloned by RT-PCR from the larva of the yellow mealworm Tenebrio molitor. The coding fragment of 339 bp encodes a protein of 112 amino acid residues and was fused to the expression vectors pET32a and pTWIN1. The resulted expression plasmids were transformed into Escherischia coli strains BL21 (DE3), ER2566, and Origami B (DE3), respectively. Several strategies were used for expression of the highly disulfide-bonded beta-helix-contained protein with the activity of antifreeze in different expression systems. A protocol for production of refolded and active T. molitor antifreeze protein in bacteria was obtained.

49 CFR 192.509 - Test requirements for pipelines to operate below 100 p.s.i. (689 kPa) gage.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 100 p.s.i. (689 kPa) gage. 192.509 Section 192.509 Transportation Other Regulations Relating to... STANDARDS Test Requirements § 192.509 Test requirements for pipelines to operate below 100 p.s.i. (689 kPa... below 100 p.s.i. (689 kPa) gage must be leak tested in accordance with the following: (a) The test...

49 CFR 192.509 - Test requirements for pipelines to operate below 100 p.s.i. (689 kPa) gage.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 100 p.s.i. (689 kPa) gage. 192.509 Section 192.509 Transportation Other Regulations Relating to... STANDARDS Test Requirements § 192.509 Test requirements for pipelines to operate below 100 p.s.i. (689 kPa... below 100 p.s.i. (689 kPa) gage must be leak tested in accordance with the following: (a) The test...

49 CFR 192.509 - Test requirements for pipelines to operate below 100 p.s.i. (689 kPa) gage.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 100 p.s.i. (689 kPa) gage. 192.509 Section 192.509 Transportation Other Regulations Relating to... STANDARDS Test Requirements § 192.509 Test requirements for pipelines to operate below 100 p.s.i. (689 kPa... below 100 p.s.i. (689 kPa) gage must be leak tested in accordance with the following: (a) The test...

49 CFR 192.509 - Test requirements for pipelines to operate below 100 p.s.i. (689 kPa) gage.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 100 p.s.i. (689 kPa) gage. 192.509 Section 192.509 Transportation Other Regulations Relating to... STANDARDS Test Requirements § 192.509 Test requirements for pipelines to operate below 100 p.s.i. (689 kPa... below 100 p.s.i. (689 kPa) gage must be leak tested in accordance with the following: (a) The test...

Suppression of polyglutamine protein toxicity by co-expression of a heat-shock protein 40 and a heat-shock protein 110

PubMed Central

Kuo, Y; Ren, S; Lao, U; Edgar, B A; Wang, T

2013-01-01

A network of heat-shock proteins mediates cellular protein homeostasis, and has a fundamental role in preventing aggregation-associated neurodegenerative diseases. In a Drosophila model of polyglutamine (polyQ) disease, the HSP40 family protein, DNAJ-1, is a superior suppressor of toxicity caused by the aggregation of polyQ containing proteins. Here, we demonstrate that one specific HSP110 protein, 70 kDa heat-shock cognate protein cb (HSC70cb), interacts physically and genetically with DNAJ-1 in vivo, and that HSC70cb is necessary for DNAJ-1 to suppress polyglutamine-induced cell death in Drosophila. Expression of HSC70cb together with DNAJ-1 significantly enhanced the suppressive effects of DNAJ-1 on polyQ-induced neurodegeneration, whereas expression of HSC70cb alone did not suppress neurodegeneration in Drosophila models of either general polyQ disease or Huntington's disease. Furthermore, expression of a human HSP40, DNAJB1, together with a human HSP110, APG-1, protected cells from polyQ-induced neural degeneration in flies, whereas expression of either component alone had little effect. Our data provide a functional link between HSP40 and HSP110 in suppressing the cytotoxicity of aggregation-prone proteins, and suggest that HSP40 and HSP110 function together in protein homeostasis control. PMID:24091676

N-terminal SKIK peptide tag markedly improves expression of difficult-to-express proteins in Escherichia coli and Saccharomyces cerevisiae.

PubMed

Ojima-Kato, Teruyo; Nagai, Satomi; Nakano, Hideo

2017-05-01

Despite advances in microbial protein expression systems, low production of proteins remains a great concern for some genes. Here we report that the insertion of a short peptide tag, consisting of Ser-Lys-Ile-Lys (SKIK), adjacent to the start codon of genes encoding difficult-to-express proteins can increase protein expression in Escherichia coli and Saccharomyces cerevisiae. Protein expression levels of a mouse monoclonal antibody (mAb), rabbit mAbs obtained from clonal B cells, and an artificially designed peptide were significantly increased simply by the addition of the SKIK tag in E. coli systems. In particular, a ∼30-fold increase in protein production was observed for the mouse mAb, and the artificially designed peptide band became detectable in sodium dodecyl sulfate-poly acrylamide gel electrophoresis after coomassie brilliant blue staining or western blotting on adding the SKIK tag. The tag also increased the expression of tagged proteins in S. cerevisiae and an E. coli cell-free protein synthesis system. Although the mechanism of high protein expression on addition of the tag is unclear, our findings offer great benefits to biotechnology research and industry. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

The N Terminus of Andes Virus L Protein Suppresses mRNA and Protein Expression in Mammalian Cells

PubMed Central

Heinemann, Patrick; Schmidt-Chanasit, Jonas

2013-01-01

Little is known about the structure and function of the 250-kDa L protein of hantaviruses, although it plays a central role in virus genome transcription and replication. When attempting to study Andes virus (ANDV) L protein in mammalian cells, we encountered difficulties. Even in a strong overexpression system, ANDV L protein could not be detected by immunoblotting. Deletion analysis revealed that the 534 N-terminal amino acid residues determine the low-expression phenotype. Inhibition of translation due to RNA secondary structures around the start codon, rapid proteasomal degradation, and reduced half-life time were excluded. However, ANDV L protein expression could be rescued upon mutation of the catalytic PD-E-K motif and further conserved residues of the putative endonuclease at the N terminus of the protein. In addition, wild-type ANDV L rather than expressible L mutants suppressed the level of L mRNA, as well as reporter mRNAs. Wild-type L protein also reduced the synthesis of cellular proteins in the high-molecular-weight range. Using expressible ANDV L mutants as a tool for localization studies, we show that L protein colocalizes with ANDV N and NSs but not Gc protein. A fraction of L protein also colocalized with the cellular processing (P) body component DCP1a. Overall, these data suggest that ANDV L protein possesses a highly active endonuclease at the N terminus suppressing the level of its own as well as heterologous mRNAs upon recombinant expression in mammalian cells. PMID:23576516

Evolved Escherichia coli strains for amplified, functional expression of membrane proteins.

PubMed

Gul, Nadia; Linares, Daniel M; Ho, Franz Y; Poolman, Bert

2014-01-09

The major barrier to the physical characterization and structure determination of membrane proteins is low protein yield and/or low functionality in recombinant expression. The enteric bacterium Escherichia coli is the most widely employed organism for producing recombinant proteins. Beside several advantages of this expression host, one major drawback is that the protein of interest does not always adopt its native conformation and may end up in large insoluble aggregates. We describe a robust strategy to increase the likelihood of overexpressing membrane proteins in a functional state. The method involves fusion in tandem of green fluorescent protein and the erythromycin resistance protein (23S ribosomal RNA adenine N-6 methyltransferase, ErmC) to the C-terminus of a target membrane protein. The fluorescence of green fluorescent protein is used to report the folding state of the target protein, whereas ErmC is used to select for increased expression. By gradually increasing the erythromycin concentration of the medium and testing different membrane protein targets, we obtained a number of evolved strains of which four (NG2, NG3, NG5 and NG6) were characterized and their genome was fully sequenced. Strikingly, each of the strains carried a mutation in the hns gene, whose product is involved in genome organization and transcriptional silencing. The degree of expression of (membrane) proteins correlates with the severity of the hns mutation, but cells in which hns was deleted showed an intermediate expression performance. We propose that (partial) removal of the transcriptional silencing mechanism changes the levels of proteins essential for the functional overexpression of membrane proteins. © 2013.

Downregulation of ATM Gene and Protein Expression in Canine Mammary Tumors.

PubMed

Raposo-Ferreira, T M M; Bueno, R C; Terra, E M; Avante, M L; Tinucci-Costa, M; Carvalho, M; Cassali, G D; Linde, S D; Rogatto, S R; Laufer-Amorim, R

2016-11-01

The ataxia telangiectasia mutated (ATM) gene encodes a protein associated with DNA damage repair and maintenance of genomic integrity. In women, ATM transcript and protein downregulation have been reported in sporadic breast carcinomas, and the absence of ATM protein expression has been associated with poor prognosis. The aim of this study was to evaluate ATM gene and protein expression in canine mammary tumors and their association with clinical outcome. ATM gene and protein expression was evaluated by reverse transcription-quantitative polymerase chain reaction and immunohistochemistry, respectively, in normal mammary gland samples (n = 10), benign mammary tumors (n = 11), nonmetastatic mammary carcinomas (n = 19), and metastatic mammary carcinomas (n = 11). Lower ATM transcript levels were detected in benign mammary tumors and carcinomas compared with normal mammary glands (P = .011). Similarly, lower ATM protein expression was observed in benign tumors (P = .0003), nonmetastatic mammary carcinomas (P < .0001), and the primary sites of metastatic carcinomas (P < .0001) compared with normal mammary glands. No significant differences in ATM gene or protein levels were detected among benign tumors and nonmetastatic and metastatic mammary carcinomas (P > .05). The levels of ATM gene or protein expression were not significantly associated with clinical and pathological features or with survival. Similar to human breast cancer, the data in this study suggest that ATM gene and protein downregulation is involved in canine mammary gland tumorigenesis. © The Author(s) 2016.

Two-station comparison of peak flows to improve flood-frequency estimates for seven streamflow-gaging stations in the Salmon and Clearwater River Basins, Central Idaho

USGS Publications Warehouse

Berenbrock, Charles

2003-01-01

Improved flood-frequency estimates for short-term (10 or fewer years of record) streamflow-gaging stations were needed to support instream flow studies by the U.S. Forest Service, which are focused on quantifying water rights necessary to maintain or restore productive fish habitat. Because peak-flow data for short-term gaging stations can be biased by having been collected during an unusually wet, dry, or otherwise unrepresentative period of record, the data may not represent the full range of potential floods at a site. To test whether peak-flow estimates for short-term gaging stations could be improved, the two-station comparison method was used to adjust the logarithmic mean and logarithmic standard deviation of peak flows for seven short-term gaging stations in the Salmon and Clearwater River Basins, central Idaho. Correlation coefficients determined from regression of peak flows for paired short-term and long-term (more than 10 years of record) gaging stations over a concurrent period of record indicated that the mean and standard deviation of peak flows for all short-term gaging stations would be improved. Flood-frequency estimates for seven short-term gaging stations were determined using the adjusted mean and standard deviation. The original (unadjusted) flood-frequency estimates for three of the seven short-term gaging stations differed from the adjusted estimates by less than 10 percent, probably because the data were collected during periods representing the full range of peak flows. Unadjusted flood-frequency estimates for four short-term gaging stations differed from the adjusted estimates by more than 10 percent; unadjusted estimates for Little Slate Creek and Salmon River near Obsidian differed from adjusted estimates by nearly 30 percent. These large differences probably are attributable to unrepresentative periods of peak-flow data collection.

Rhythmic expression of DEC2 protein in vitro and in vivo.

PubMed

Sato, Fuyuki; Muragaki, Yasuteru; Kawamoto, Takeshi; Fujimoto, Katsumi; Kato, Yukio; Zhang, Yanping

2016-06-01

Basic helix-loop-helix (bHLH) transcription factor DEC2 (bHLHE41/Sharp1) is one of the clock genes that show a circadian rhythm in various tissues. DEC2 regulates differentiation, sleep length, tumor cell invasion and apoptosis. Although studies have been conducted on the rhythmic expression of DEC2 mRNA in various tissues, the precise molecular mechanism of DEC2 expression is poorly understood. In the present study, we examined whether DEC2 protein had a rhythmic expression. Western blot analysis for DEC2 protein revealed a rhythmic expression in mouse liver, lung and muscle and in MCF-7 and U2OS cells. In addition, AMP-activated protein kinase (AMPK) activity (phosphorylation of AMPK) in mouse embryonic fibroblasts (MEFs) exhibited a rhythmic expression under the condition of medium change or glucose-depleted medium. However, the rhythmic expression of DEC2 in MEF gradually decreased in time under these conditions. The medium change affected the levels of DEC2 protein and phosphorylation of AMPK. In addition, the levels of DEC2 protein showed a rhythmic expression in vivo and in MCF-7 and U2OS cells. The results showed that the phosphorylation of AMPK immunoreactivity was strongly detected in the liver and lung of DEC2 knockout mice compared with that of wild-type mice. These results may provide new insights into rhythmic expression and the regulation between DEC2 protein and AMPK activity.

Novel isoprenylated proteins identified by an expression library screen.

PubMed

Biermann, B J; Morehead, T A; Tate, S E; Price, J R; Randall, S K; Crowell, D N

1994-10-14

Isoprenylated proteins are involved in eukaryotic cell growth and signal transduction. The protein determinant for prenylation is a short carboxyl-terminal motif containing a cysteine, to which the isoprenoid is covalently attached via thioether linkage. To date, isoprenylated proteins have almost all been identified by demonstrating the attachment of an isoprenoid to previously known proteins. Thus, many isoprenylated proteins probably remain undiscovered. To identify novel isoprenylated proteins for subsequent biochemical study, colony blots of a Glycine max cDNA expression library were [3H]farnesyl-labeled in vitro. Proteins identified by this screen contained several different carboxyl termini that conform to consensus farnesylation motifs. These proteins included known farnesylated proteins (DnaJ homologs) and several novel proteins, two of which contained six or more tandem repeats of a hexapeptide having the consensus sequence (E/G)(G/P)EK(P/K)K. Thus, plants contain a diverse array of genes encoding farnesylated proteins, and our results indicate that fundamental differences in the identities of farnesylated proteins may exist between plants and other eukaryotes. Expression library screening by direct labeling can be adapted to identify isoprenylated proteins from other organisms, as well as proteins with other post-translational modifications.

Genome engineering for improved recombinant protein expression in Escherichia coli.

PubMed

Mahalik, Shubhashree; Sharma, Ashish K; Mukherjee, Krishna J

2014-12-19

A metabolic engineering perspective which views recombinant protein expression as a multistep pathway allows us to move beyond vector design and identify the downstream rate limiting steps in expression. In E.coli these are typically at the translational level and the supply of precursors in the form of energy, amino acids and nucleotides. Further recombinant protein production triggers a global cellular stress response which feedback inhibits both growth and product formation. Countering this requires a system level analysis followed by a rational host cell engineering to sustain expression for longer time periods. Another strategy to increase protein yields could be to divert the metabolic flux away from biomass formation and towards recombinant protein production. This would require a growth stoppage mechanism which does not affect the metabolic activity of the cell or the transcriptional or translational efficiencies. Finally cells have to be designed for efficient export to prevent buildup of proteins inside the cytoplasm and also simplify downstream processing. The rational and the high throughput strategies that can be used for the construction of such improved host cell platforms for recombinant protein expression is the focus of this review.

Inhibition of interferon-inducible MxA protein expression by hepatitis B virus capsid protein.

PubMed

Rosmorduc, O; Sirma, H; Soussan, P; Gordien, E; Lebon, P; Horisberger, M; Bréchot, C; Kremsdorf, D

1999-05-01

Chronic hepatitis B treatment has been significantly improved by interferon (IFN) treatment. However, some studies have suggested that hepatitis B virus (HBV) might have a direct effect on the resistance to IFN. Defective particles, generated by spliced HBV RNA and associated with chronic hepatitis B, have been previously characterized; expression of these particles leads to cytoplasmic accumulation of the capsid protein. The aim of this study was to investigate the role of these defective genomes in IFN resistance. The global antiviral activity of IFN was studied by virus yield reduction assays, the expression of three IFN-induced antiviral proteins was analysed by Western blotting and confocal microscopy, and the regulation of MxA gene expression was studied by Northern blotting and the luciferase assay, in Huh7 cells transfected with a complete or the defective HBV genome. Results showed that the expression of the defective genome reduces the antiviral activity of IFN and that this modulation involves a selective inhibition of MxA protein induction by the HBV capsid protein. Our results also show the trans-suppressive effect of the HBV capsid on the MxA promoter, which might participate in this phenomenon. In conclusion, this study shows a direct interplay between the IFN-sensitive pathway and the capsid protein and might implicate this defective HBV genome in virus persistence.

Utilizing Photogrammetry and Strain Gage Measurement to Characterize Pressurization of an Inflatable Module

NASA Technical Reports Server (NTRS)

Valle, Gerard D.; Selig, Molly; Litteken, Doug; Oliveras, Ovidio

2012-01-01

This paper documents the integration of a large hatch penetration into an inflatable module. This paper also documents the comparison of analytical load predictions with measured results utilizing strain measurement. Strain was measured by utilizing photogrammetric measurement and through measurement obtained from strain gages mounted to selected clevises that interface with the structural webbings. Bench testing showed good correlation between strain measurement obtained from an extensometer and photogrammetric measurement especially after the fabric has transitioned through the low load/high strain region of the curve. Test results for the full-scale torus showed mixed results in the lower load and thus lower strain regions. Overall strain, and thus load, measured by strain gages and photogrammetry tracked fairly well with analytical predictions. Methods and areas of improvements are discussed.

Transforming Lepidopteran Insect Cells for Improved Protein Processing and Expression

USDA-ARS?s Scientific Manuscript database

The lepidopteran insect cells used with the baculovirus expression vector system (BEVS) are capable of synthesizing and accurately processing foreign proteins. However, proteins expressed in baculovirus-infected cells often fail to be completely processed, or are not processed in a manner that meet...

Techniques for estimating monthly mean streamflow at gaged sites and monthly streamflow duration characteristics at ungaged sites in central Nevada

USGS Publications Warehouse

Hess, G.W.; Bohman, L.R.

1996-01-01

Techniques for estimating monthly mean streamflow at gaged sites and monthly streamflow duration characteristics at ungaged sites in central Nevada were developed using streamflow records at six gaged sites and basin physical and climatic characteristics. Streamflow data at gaged sites were related by regression techniques to concurrent flows at nearby gaging stations so that monthly mean streamflows for periods of missing or no record can be estimated for gaged sites in central Nevada. The standard error of estimate for relations at these sites ranged from 12 to 196 percent. Also, monthly streamflow data for selected percent exceedence levels were used in regression analyses with basin and climatic variables to determine relations for ungaged basins for annual and monthly percent exceedence levels. Analyses indicate that the drainage area and percent of drainage area at altitudes greater than 10,000 feet are the most significant variables. For the annual percent exceedence, the standard error of estimate of the relations for ungaged sites ranged from 51 to 96 percent and standard error of prediction for ungaged sites ranged from 96 to 249 percent. For the monthly percent exceedence values, the standard error of estimate of the relations ranged from 31 to 168 percent, and the standard error of prediction ranged from 115 to 3,124 percent. Reliability and limitations of the estimating methods are described.

Optimised 'on demand' protein arraying from DNA by cell free expression with the 'DNA to Protein Array' (DAPA) technology.

PubMed

Schmidt, Ronny; Cook, Elizabeth A; Kastelic, Damjana; Taussig, Michael J; Stoevesandt, Oda

2013-08-02

We have previously described a protein arraying process based on cell free expression from DNA template arrays (DNA Array to Protein Array, DAPA). Here, we have investigated the influence of different array support coatings (Ni-NTA, Epoxy, 3D-Epoxy and Polyethylene glycol methacrylate (PEGMA)). Their optimal combination yields an increased amount of detected protein and an optimised spot morphology on the resulting protein array compared to the previously published protocol. The specificity of protein capture was improved using a tag-specific capture antibody on a protein repellent surface coating. The conditions for protein expression were optimised to yield the maximum amount of protein or the best detection results using specific monoclonal antibodies or a scaffold binder against the expressed targets. The optimised DAPA system was able to increase by threefold the expression of a representative model protein while conserving recognition by a specific antibody. The amount of expressed protein in DAPA was comparable to those of classically spotted protein arrays. Reaction conditions can be tailored to suit the application of interest. DAPA represents a cost effective, easy and convenient way of producing protein arrays on demand. The reported work is expected to facilitate the application of DAPA for personalized medicine and screening purposes. Copyright © 2013 Elsevier B.V. All rights reserved.

[Eukaryotic Expression and Immunogenic Research of Recombination Ebola Virus Membrane Protein Gp-Fc].

PubMed

Zhang, Xiaoguang; Yang, Ren; Wang, Jiao; Wang, Xuan; Hou, Mieling; An, Lina; Zhu, Ying; Cao, Yuxi; Zeng, Yi

2016-01-01

We used 293 cells to express the recombinant membrane protein of the Ebola virus. Then, the immunogenicity of the recombinant protein was studied by immunized BALB/c mice. According to the codon use frequency of humans, the gene encoding the extracellular domain of the Ebola virus membrane protein was optimized, synthesized, and inserted into the eukaryotic expression plasmid pXG-Fc to construct the human IgG Fc and Ebola GP fusion protein expression plasmid pXG-modGP-Fc. To achieve expression, the fusion protein expression vector was transfected into high-density 293 cells using transient transfection technology. The recombinant protein was purified by protein A affinity chromatography. BALB/c mice were immunized with the purified fusion protein, and serum antibody titers evaluated by an indirect enzyme-linked immunosorbent assay (ELISA). Purification and analyses of the protein revealed that the eukaryotic expression vector could express the recombinant protein GP-Fc effectively, and that the recombinant protein in the supernatant of the cell culture was present as a dimer. After immunization with the purified recombinant protein, a high titer of antigen-specific IgG could be detected in the serum of immunized mice by indirect ELISA, showing that the recombinant protein had good immunogenicity. These data suggest that we obtained a recombinant protein with good immunogenicity. Our study is the basis for development of a vaccine against the Ebola virus and for screening of monoclonal antibodies.

A Generic Protocol for Intracellular Expression of Recombinant Proteins in Bacillus subtilis.

PubMed

Phan, Trang; Huynh, Phuong; Truong, Tuom; Nguyen, Hoang

2017-01-01

Bacillus subtilis (B. subtilis) is a potential and attractive host for the production of recombinant proteins. Different expression systems for B. subtilis have been developed recently, and various target proteins have been recombinantly synthesized and purified using this host. In this chapter, we introduce a generic protocol to express a recombinant protein in B. subtilis. It includes protocols for (1) using our typical expression vector (plasmid pHT254) to introduce a target gene, (2) transformation of the target vector into B. subtilis, and (3) evaluation of the actual expression of a recombinant protein.

Expression of membrane-associated proteins within single emulsion cell facsimiles.

PubMed

Chanasakulniyom, Mayuree; Martino, Chiara; Paterson, David; Horsfall, Louise; Rosser, Susan; Cooper, Jonathan M

2012-07-07

MreB is a structural membrane-associated protein which is one of the key components of the bacterial cytoskeleton. Although it plays an important role in shape maintenance of rod-like bacteria, the understanding of its mechanism of action is still not fully understood. This study shows how segmented flow and microdroplet technology can be used as a new tool for biological in vitro investigation of this protein. In this paper, we demonstrate cell-free expression in a single emulsion system to express red fluorescence protein (RFP) and MreB linked RFP (MreB-RFP). We follow the aggregation and localisation of the fusion protein MreB-RFP in this artificial cell-like environment. The expression of MreB-RFP in single emulsion droplets leads to the formation of micrometer-scale protein patches distributed at the water/oil interface.

Expression and Localization of Plant Protein Disulfide Isomerase.

PubMed Central

Shorrosh, B. S.; Subramaniam, J.; Schubert, K. R.; Dixon, R. A.

1993-01-01

A cDNA clone encoding a putative protein disulfide isomerase (PDI, EC 5.3.4.1) from alfalfa (Medicago sativa L.) was expressed in Escherichia coli cells, and an antiserum was raised against the expressed PDI-active protein. The antiserum recognized a protein of approximately 60 kD in extracts from alfalfa, soybean, and tobacco roots and stems. Levels of this protein remained relatively constant on exposure of alfalfa cell suspension cultures to the protein glycosylation inhibitor tunicamycin, whereas a slightly lower molecular mass form, also detected by the antiserum, was induced by this treatment. A lower molecular mass form of PDI was also observed in roots of alfalfa seedlings during the first 5 weeks after germination. PDI levels increased in developing soybean seeds up to 17 d after fertilization and then declined. Tissue print immunoblots revealed highest levels of PDI protein in the cambial tissues of soybean stems and petioles and in epidermal, subepidermal, cortical, and pith tissues of stems of alfalfa and tobacco. Immunogold electron microscopy confirmed the localization of PDI to the endoplasmic reticulum in soybean root nodules. PMID:12231974

Common and specific signatures of gene expression and protein-protein interactions in autoimmune diseases.

PubMed

Tuller, T; Atar, S; Ruppin, E; Gurevich, M; Achiron, A

2013-03-01

The aim of this study is to understand intracellular regulatory mechanisms in peripheral blood mononuclear cells (PBMCs), which are either common to many autoimmune diseases or specific to some of them. We incorporated large-scale data such as protein-protein interactions, gene expression and demographical information of hundreds of patients and healthy subjects, related to six autoimmune diseases with available large-scale gene expression measurements: multiple sclerosis (MS), systemic lupus erythematosus (SLE), juvenile rheumatoid arthritis (JRA), Crohn's disease (CD), ulcerative colitis (UC) and type 1 diabetes (T1D). These data were analyzed concurrently by statistical and systems biology approaches tailored for this purpose. We found that chemokines such as CXCL1-3, 5, 6 and the interleukin (IL) IL8 tend to be differentially expressed in PBMCs of patients with the analyzed autoimmune diseases. In addition, the anti-apoptotic gene BCL3, interferon-γ (IFNG), and the vitamin D receptor (VDR) gene physically interact with significantly many genes that tend to be differentially expressed in PBMCs of patients with the analyzed autoimmune diseases. In general, similar cellular processes tend to be differentially expressed in PBMC in the analyzed autoimmune diseases. Specifically, the cellular processes related to cell proliferation (for example, epidermal growth factor, platelet-derived growth factor, nuclear factor-κB, Wnt/β-catenin signaling, stress-activated protein kinase c-Jun NH2-terminal kinase), inflammatory response (for example, interleukins IL2 and IL6, the cytokine granulocyte-macrophage colony-stimulating factor and the B-cell receptor), general signaling cascades (for example, mitogen-activated protein kinase, extracellular signal-regulated kinase, p38 and TRK) and apoptosis are activated in most of the analyzed autoimmune diseases. However, our results suggest that in each of the analyzed diseases, apoptosis and chemotaxis are activated via

Parasitization by Scleroderma guani influences protein expression in Tenebrio molitor pupae.

PubMed

Zhu, Jia-Ying; Wu, Guo-Xing; Ze, Sang-Zi; Stanley, David W; Yang, Bin

2014-07-01

Ectoparasitoid wasps deposit their eggs onto the surface and inject venom into their hosts. Venoms are chemically complex and they exert substantial impact on hosts, including permanent or temporary paralysis and developmental arrest. These visible venom effects are due to changes in expression of genes encoding physiologically relevant proteins. While the influence of parasitization on gene expression in several lepidopterans has been reported, the molecular details of parasitoid/beetle relationships remain mostly unknown. This shortcoming led us to pose the hypothesis that envenomation by the ectoparasitic ant-like bethylid wasp Scleroderma guani leads to changes in protein expression in the yellow mealworm beetle Tenebrio molitor. We tested our hypothesis by comparing the proteomes of non-parasitized and parasitized host pupae using iTRAQ-based proteomics. We identified 41 proteins that were differentially expressed (32↑- and 9↓-regulated) in parasitized pupae. We assigned these proteins to functional categories, including immunity, stress and detoxification, energy metabolism, development, cytoskeleton, signaling and others. We recorded parallel changes in mRNA levels and protein abundance in 14 selected proteins following parasitization. Our findings support our hypothesis by documenting changes in protein expression in parasitized hosts. Copyright © 2014 Elsevier Ltd. All rights reserved.

Insect cells-baculovirus system for the production of difficult to express proteins.

PubMed

Osz-Papai, Judit; Radu, Laura; Abdulrahman, Wassim; Kolb-Cheynel, Isabelle; Troffer-Charlier, Nathalie; Birck, Catherine; Poterszman, Arnaud

2015-01-01

The production of sufficient quantities of homogenous protein not only is an essential prelude for structural investigations but also represents a rate-limiting step for many human functional studies. Although technologies for expression of recombinant proteins and complexes have been improved tremendously, in many cases, protein production remains a challenge and can be associated with considerable investment. This chapter describes simple and efficient protocols for expression screening and optimization of protein production in insect cells using the baculovirus expression system. We describe the procedure, starting from the cloning of a gene of interest into an expression transfer baculovirus vector, followed by generation of the recombinant virus by homologous recombination, evaluation of protein expression, and scale-up. Handling of insect cell cultures and preparation of bacmid for co-transfection are also detailed.

Strain Gage Loads Calibration Testing with Airbag Support for the Gulfstream III SubsoniC Research Aircraft Testbed (SCRAT)

NASA Technical Reports Server (NTRS)

Lokos, William; Miller, Eric; Hudson, Larry; Holguin, Andrew; Neufeld, David; Haraguchi, Ronnie

2015-01-01

This paper describes the design and conduct of the strain gage load calibration ground test of the SubsoniC Research Aircraft Testbed, Gulfstream III aircraft, and the subsequent data analysis and its results. The goal of this effort was to create and validate multi-gage load equations for shear force, bending moment, and torque for two wing measurement stations. For some of the testing the aircraft was supported by three air bags in order to isolate the wing structure from extraneous load inputs through the main landing gear. Thirty-two strain gage bridges were installed on the left wing. Hydraulic loads were applied to the wing lower surface through a total of 16 load zones. Some dead weight load cases were applied to the upper wing surface using shot bags. Maximum applied loads reached 54,000 pounds.

Evaluation of a Quartz Bourdon Pressure Gage of Wind Tunnel Mach Number Control System Application

NASA Technical Reports Server (NTRS)

Chapin, W. G.

1986-01-01

A theoretical and experimental study was undertaken to determine the feasibility of using the National Transonic Facility's high accuracy Mach number measurement system as part of a closed loop Mach number control system. The theoretical and experimental procedures described are applicable to the engineering design of pressure control systems. The results show that the dynamic response characteristics of the NTF Mach number gage (a Ruska DDR-6000 quartz absolute pressure gage) coupled to a typical length of pressure tubing were only marginally acceptable within a limited range of the facility's total pressure envelope and could not be used in the Mach number control system.

Novel leukocyte protein, Trojan, differentially expressed during thymocyte development.

PubMed

Petrov, Petar; Motobu, Maki; Salmi, Jussi; Uchida, Tatsuya; Vainio, Olli

2010-04-01

"Trojan" is a novel cell surface protein, discovered from chicken embryonic thymocytes on the purpose to identify molecules involved in T cell differentiation. The molecule is predicted as a type I transmembrane protein having a Sushi and two fibronectin type III domains and a pair of intracellular phosphorylation sites. Its transcript expression is specific for lymphoid tissues and the presence of the protein on the surface of recirculating lymphocytes and macrophages was confirmed by immunofluorescence analysis. In thymus, about half of the double negative (CD4(-) CD8(-)) and CD8 single positive and the majority of CD4 single positive cells express Trojan with a relatively high intensity. However, only a minority of the double positive (CD4(+) CD8(+)) cells are positive for Trojan. This expression pattern, similar to that of some proteins with anti-apoptotic and function, like IL-7Ralpha, makes Trojan an attractive candidate of having an anti-apoptotic role. Copyright 2010 Elsevier Ltd. All rights reserved.

Rift Valley Fever Virus Structural and Nonstructural Proteins: Recombinant Protein Expression and Immunoreactivity Against Antisera from Sheep

PubMed Central

Faburay, Bonto; Wilson, William; McVey, D. Scott; Drolet, Barbara S.; Weingartl, Hana; Madden, Daniel; Young, Alan; Ma, Wenjun

2013-01-01

Abstract The Rift Valley fever virus (RVFV) encodes the structural proteins nucleoprotein (N), aminoterminal glycoprotein (Gn), carboxyterminal glycoprotein (Gc), and L protein, 78-kD, and the nonstructural proteins NSm and NSs. Using the baculovirus system, we expressed the full-length coding sequence of N, NSs, NSm, Gc, and the ectodomain of the coding sequence of the Gn glycoprotein derived from the virulent strain of RVFV ZH548. Western blot analysis using anti-His antibodies and monoclonal antibodies against Gn and N confirmed expression of the recombinant proteins, and in vitro biochemical analysis showed that the two glycoproteins, Gn and Gc, were expressed in glycosylated form. Immunoreactivity profiles of the recombinant proteins in western blot and in indirect enzyme-linked immunosorbent assay against a panel of antisera obtained from vaccinated or wild type (RVFV)-challenged sheep confirmed the results obtained with anti-His antibodies and demonstrated the suitability of the baculo-expressed antigens for diagnostic assays. In addition, these recombinant proteins could be valuable for the development of diagnostic methods that differentiate infected from vaccinated animals (DIVA). PMID:23962238

Screening Fusion Tags for Improved Recombinant Protein Expression in E. coli with the Expresso® Solubility and Expression Screening System.

PubMed

Steinmetz, Eric J; Auldridge, Michele E

2017-11-01

The simplicity, speed, and low cost of bacterial culture make E. coli the system of choice for most initial trials of recombinant protein expression. However, many heterologous proteins are either poorly expressed in bacteria, or are produced as incorrectly folded, insoluble aggregates that lack the activity of the native protein. In many cases, fusion to a partner protein can allow for improved expression and/or solubility of a difficult target protein. Although several different fusion partners have gained favor, none are universally effective, and identifying the one that best improves soluble expression of a given target protein is an empirical process. This unit presents a strategy for parallel screening of fusion partners for enhanced expression or solubility. The Expresso® Solubility and Expression Screening System includes a panel of seven distinct fusion partners and utilizes an extremely simple cloning strategy to enable rapid screening and identification of the most effective fusion partner. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

Evaluation of metal-foil strain gages for cryogenic application in magnetic fields

DOE Office of Scientific and Technical Information (OSTI.GOV)

Freynik, H.S. Jr.; Roach, D.R.; Deis, D.W.

1977-07-08

The requirement for the design and construction of large superconducting magnet systems for fusion research has raised a number of new questions regarding the properties of composite superconducting conductors. One of these, the effect of mechanical stress on the current-carrying capacity of Nb/sub 3/Sn, is of major importance in determining the feasibility of constructing large magnets with this material. A typical experiment for determining such data involves the measurement of critical current versus magnetic field while the conductor is being mechanically strained to various degrees. Techniques are well developed for the current and field measurements, but much less so formore » the accurate measurement of strain at liquid-helium temperature in a high magnetic field. A study was made of commercial, metal-foil strain gages for use under these conditions. The information developed can also be applied to the use of strain gages as diagnostic tools in superconducting magnets.« less

The accuracy of mean-growth estimates made with dial-gage dendrometers

Treesearch

Barton M. Blum; Dale S. Solomon

1966-01-01

In recent years a great deal of interest has been focused on tree growth as measured with relatively precise instruments over short periods of time. These growth measurements are usually made with vernier-band dendrometers similar to those described by Hall (1944) and Liming (1957), or dial-gage dendrometers similar to those described by Reineke (1932) and Daubenmire (...

Expression of APG-2 protein, a member of the heat shock protein 110 family, in developing rat brain.

PubMed

Okui, M; Ito, F; Ogita, K; Kuramoto, N; Kudoh, J; Shimizu, N; Ide, T

2000-01-01

APG-2 protein is a member of the heat shock protein 110 family, and it is thought to play an important role in the maintenance of neuronal functions under physiological and stress conditions. However, neither the tissue-distribution of APG-2 protein nor developmental change of its expression has been studied at the protein level. Therefore, we generated an antiserum against APG-2 protein and studied expression of this protein in rat brain and other tissues by use of the Western blot method. The results showed a high expression of APG-2 protein in various regions of the central nervous system (cerebral cortex, hippocampus, striatum, midbrain, hypothalamus, cerebellum, medulla pons, and spinal cord) throughout the entire postnatal stage. Similarly, a high level of APG-2 protein was detected in the whole brain of rat embryos and in adult rat tissues such as liver, lung, spleen, and kidney. In contrast, its expression in heart was high at postnatal days 1 and 3, but thereafter drastically decreased to a low level. Furthermore, APG-2 protein was detected in neuronal primary cultures prepared from rat cerebral cortex, and its level did not change notably during neuronal differentiation. These results show that APG-2 protein is constitutively expressed in various tissues and also in neuronal cells throughout the entire embryonic and postnatal period. suggesting that it might play an important role in these tissues under non-stress conditions.

Marker Protein Expression Combined With Expression Heterogeneity is a Powerful Indicator of Malignancy in Acral Lentiginous Melanomas.

PubMed

Cintra Lopes Carapeto, Fernando; Neves Comodo, Andréia; Germano, Andressa; Pereira Guimarães, Daiane; Barcelos, Denise; Fernandes, Mariana; Landman, Gilles

2017-02-01

Samples of acral lentiginous melanomas (ALMs) were obtained from the Department of Pathology at Escola Paulista de Medicina-Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil. Demographic, clinical, and follow-up data were obtained from the charts of Hospital São Paulo. From 2 tissue microarrays containing 60 nevi and quadruplicate samples of ≥1.0-mm of 49 ALM, sections were stained to evaluate SCF, KIT, BRAF, CYCLIND1, MYC, and PTEN immunohistochemical protein expression. Nevi and ALM from 2006 to 2010 were reviewed and collected. All specimens were in the vertical growth phase, and histopathological parameters indicated that tumors were at an advanced stage at diagnosis. Average tumor thickness was 6.95 mm, 63% were ulcerated, average mitotic index was 5 mitotic cells per mm, and 43% were at Clark's level V. Compared with nevi, the χ test showed that ALM significantly correlated with SCF protein expression (P = 0.001) and expression heterogeneity (P < 0.000). Similar findings were observed for KIT (P = 0.005, P = 0.003, respectively), MYC (P < 0.000, P < 0.000), and PTEN (P = 0.005, P < 0.000). Malignancy did not correlate with BRAF and CYCLIN D1 expression (P = 0.053 and P = 0.259, respectively), but it did significantly correlate with their heterogeneous expression (P < 0.000, P = 0.024, respectively). Combined protein expression had an odds ratio of greater malignancy when BRAF and MYC were positive and/or heterogeneously expressed (OR of 78 and 95, respectively). We show that marker protein expression, when combined with heterogeneous expression as shown by immunohistochemistry, is a powerful indicator of malignancy in ALMs, especially, when protein pairs are combined.

Exploiting translational coupling for the selection of cells producing toxic recombinant proteins from expression vectors.

PubMed

Tagliavia, Marcello; Cuttitta, Angela

2016-01-01

High rates of plasmid instability are associated with the use of some expression vectors in Escherichia coli, resulting in the loss of recombinant protein expression. This is due to sequence alterations in vector promoter elements caused by the background expression of the cloned gene, which leads to the selection of fast-growing, plasmid-containing cells that do not express the target protein. This phenomenon, which is worsened when expressing toxic proteins, results in preparations containing very little or no recombinant protein, or even in clone loss; however, no methods to prevent loss of recombinant protein expression are currently available. We have exploited the phenomenon of translational coupling, a mechanism of prokaryotic gene expression regulation, in order to select cells containing plasmids still able to express recombinant proteins. Here we designed an expression vector in which the cloned gene and selection marker are co-expressed. Our approach allowed for the selection of the recombinant protein-expressing cells and proved effective even for clones encoding toxic proteins.

Prognostic relevance of Centromere protein H expression in esophageal carcinoma.

PubMed

Guo, Xian-Zhi; Zhang, Ge; Wang, Jun-Ye; Liu, Wan-Li; Wang, Fang; Dong, Ju-Qin; Xu, Li-Hua; Cao, Jing-Yan; Song, Li-Bing; Zeng, Mu-Sheng

2008-08-13

Many kinetochore proteins have been shown to be associated with human cancers. The aim of the present study was to clarify the expression of Centromere protein H (CENP-H), one of the fundamental components of the human active kinetochore, in esophageal carcinoma and its correlation with clinicopathological features. We examined the expression of CENP-H in immortalized esophageal epithelial cells as well as in esophageal carcinoma cells, and in 12 cases of esophageal carcinoma tissues and the paired normal esophageal tissues by RT-PCR and Western blot analysis. In addition, we analyzed CENP-H protein expression in 177 clinicopathologically characterized esophageal carcinoma cases by immunohistochemistry. Statistical analyses were applied to test for prognostic and diagnostic associations. The level of CENP-H mRNA and protein were higher in the immortalized cells, cancer cell lines and most cancer tissues than in normal control tissues. Immunohistochemistry showed that CENP-H was expressed in 127 of 171 ESCC cases (74.3%) and in 3 of 6 esophageal adenocarcinoma cases (50%). Statistical analysis of ESCC cases showed that there was a significant difference of CENP-H expression in patients categorized according to gender (P = 0.013), stage (P = 0.023) and T classification (P = 0.019). Patients with lower CENP-H expression had longer overall survival time than those with higher CENP-H expression. Multivariate analysis suggested that CENP-H expression was an independent prognostic marker for esophageal carcinoma patients. A prognostic value of CENP-H was also found in the subgroup of T3 approximately T4 and N0 tumor classification. Our results suggest that CENP-H protein is a valuable marker of esophageal carcinoma progression. CENP-H might be used as a valuable prognostic marker for esophageal carcinoma patients.

Calcium affecting protein expression in longan under simulated acid rain stress.

PubMed

Pan, Tengfei; Li, Yongyu; Ma, Cuilan; Qiu, Dongliang

2015-08-01

Longan (Dimocarpus longana Lour. cv. Wulongling) of uniform one-aged seedlings grown in pots were selected to study specific proteins expressed in leaves under simulated acid rain (SiAR) stress and exogenous Ca(2+) regulation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results showed that there was a protein band specifically expressed under SiAR of pH 2.5 stress for 15 days with its molecular weight of about 23 kD. A 17 kD protein band specifically expressed after SiAR stress 5 days. Compared with pH 2.5, the pH 3.5 of SiAR made a less influence to protein expression. Two-dimensional electrophoresis (2-DE) results showed that six new specific proteins including C4 (20.2 kD pI 6.0), F (24 kD pI 6.35), B3 (22.3 kD pI 6.35), B4 (23.5 kD pI 6.5), C5 (21.8 kD pI 5.6), and C6 (20.2 kD pI 5.6) specifically expressed. C4 always expressed during SiAR stress. F expressed under the stress of pH 2.5 for 15 days and expressed in all pH SiAR stress for 20 days. The expression of proteins including B3, C5, and C6 was related to pH value and stress intensity of SiAR. The expression of B4 resulted from synergistic effects of SiAR and Ca. The expression of G1 (Mr 19.3 kD, pI 4.5), G2 (Mr 17.8 kD, pI 4.65), G3 (Mr 16.6 kD, pI 4.6), and G4 (Mr 14.7 kD, pI 4.4) enhanced under the treatment of 5 mM ethylene glycol tetraacetic acid (EGTA) and 2 mM chlorpromazine (CPZ). These proteins showed antagonistic effects and might be relative to the Ca-calmodulin (Ca-CaM) system of longan in response to SiAR stress.

Heterogeneity of proteins expressed by Brazilian Sporothrix schenckii isolates.

PubMed

Fernandes, Geisa Ferreira; Do Amaral, Cristiane Candida; Sasaki, Alexandre; Godoy, Patrício Martinez; De Camargo, Zoilo Pires

2009-12-01

The profiles of proteins present in the exoantigens of Brazilian Sporothrix schenckii isolates were studied and compared by electrophoresis (SDS-PAGE). Thirteen isolates from five different regions of Brazil (1,000 to 2,000 km apart) and ten from a more limited region (200 to 400 km apart within the state of São Paulo) were cultured in Sabouraud, M199 and minimum (MM) media. Qualitative and quantitative differences in the expression of proteins, which varied according to the medium and the isolate, were observed. Fractions with the same MW but varying in intensity were detected, as well as fractions present in 1 isolate but absent in others. Dendrograms were constructed and isolates grouped based on the fractions obtained, irrespective of the intensity. The results showed that Brazilian S. schenckii isolates express different protein profiles, a feature also present in isolates from a more restricted region. The exoantigens were found to have a maximum of 15 protein fractions, ranging in MW from 19-220 KDaltons depending on the medium used for the cultures. These data show the great heterogeneity of Brazilian S. schenckii protein expression.

Strain gage attachment by spot welding reduces the fatigue strength of Ti-6Al-4V, Rene 41, and Inconel X

NASA Technical Reports Server (NTRS)

Imig, L. A.

1972-01-01

Fatigue tests were conducted with constant-amplitude axial stresses in the ratio of minimum to maximum stress of 0.05 (R=0.05). Specimens with and without strain gages were tested at 21 C, and superalloy specimens with and without strain gages were tested at 21 C and 815 C.

Recombinant protein expression in Escherichia coli: advances and challenges

PubMed Central

Rosano, Germán L.; Ceccarelli, Eduardo A.

2014-01-01

Escherichia coli is one of the organisms of choice for the production of recombinant proteins. Its use as a cell factory is well-established and it has become the most popular expression platform. For this reason, there are many molecular tools and protocols at hand for the high-level production of heterologous proteins, such as a vast catalog of expression plasmids, a great number of engineered strains and many cultivation strategies. We review the different approaches for the synthesis of recombinant proteins in E. coli and discuss recent progress in this ever-growing field. PMID:24860555

Transient expression and cellular localization of recombinant proteins in cultured insect cells

USDA-ARS?s Scientific Manuscript database

Heterologous protein expression systems are used for production of recombinant proteins, interpretation of cellular trafficking/localization, and for the determination of biochemical function of proteins at the sub-organismal level. Although baculovirus expression systems are increasingly used for ...

Expression of Lipid Metabolism-Related Proteins in Metastatic Breast Cancer.

PubMed

Jung, Yoon Yang; Kim, Hye Min; Koo, Ja Seung

2015-01-01

The tumor biology of metastatic breast cancers differ according to the metastatic sites, and the features of cancer metabolism may also be different. The aim of this study is to investigate the expression of lipid metabolism-related proteins in metastatic breast cancer according to metastatic site and discuss the clinical significance thereof. Immunohistochemical staining for lipid metabolism-related proteins [fatty acid synthase (FASN), hormone-sensitive lipase (HSL), carnitine palmitoyltransferase IA (CPT-1A), acyl-CoA oxidase 1 (ACOX1), fatty acid binding protein 4 (FABP4,) and perilipin 1 (PLIN1)] was performed using a tissue microarray of 149 cases of metastatic breast cancer (bone metastasis = 39, brain metastasis = 37, liver metastasis = 21, and lung metastasis = 52). The expression levels of ACOX1 (p = 0.009) and FASN (p = 0.007) varied significantly according to metastatic site, with the highest expression in brain metastasis and the lowest expression in liver metastasis. ACOX1 positivity (p = 0.005) and FASN positivity (p = 0.003) correlated with HER-2 positivity. The expression of FASN was significantly higher in HER-2 type breast cancer, and lower in luminal A and TNBC type breast cancer (p<0.001). Among lipid metabolism-related proteins, PLIN1 positivity was found to be an independent poor prognostic factor on multivariate analysis (Hazard ratio: 4.979, 95% CI: 1.054-22.59, p = 0.043). Different expression levels of lipid metabolism-related proteins were observed according to metastatic site. The expression of ACOX1 and FASN was highest in brain metastasis. These results suggest that the metastatic site should be considered when using lipid metabolism inhibitors for targeted therapy.

A Double-Primary Dead-Weight Tester for Pressures (35-175) kPa in Gage Mode.

PubMed

Jain, Kamlesh; Cen, Yueqin; Bowers, Walter J; Schmidt, James W

2003-01-01

Primary pressure standards in the atmospheric pressure range are often established using mercury manometers. Less frequently, controlled-clearance dead-weight testers in which one component (normally the piston) has been dimensionally measured have also been used. Recent advances in technology on two fronts i) the fabrication of large-diameter pistons and cylinders with good geometry; and ii) the ability to measure the dimensions of these components, have allowed some dead-weight testers at NIST to approach total relative uncertainties (k = 2) in dimensionally-derived effective areas near 5 × 10(-6). This paper describes a single piston/cylinder assembly (NIST-PG201WC/WC) that serves as both a primary gage in which both piston and cylinder are measured dimensionally and a controlled-clearance primary gage (employing the Heydemann-Welch method). Thus it allows some previous assumptions about the modeling of dead-weight testers to be checked. For the gage described in this paper the piston/cylinder clearance obtained from the two analyses have relative differences of 4 × 10(-6) to 7 × 10(-6) over the pressure range 35 kPa to 175 kPa. Some implications of these results will be discussed. From the dimensional characterizations and auxiliary measurements we have determined that the effective area for this gauge at 20 °C is: [Formula: see text]where P is the system pressure and P J is a control pressure. The estimated relative uncertainty in effective area is 8.2 × 10(-6) +1.4 × 10(-11) P/Pa (k = 2). The temperature coefficient for the area was measured and found to be (9.06 ± 0.04) × 10(-6)/K. Thus using the gage at a reference temperature of 23 °C yields an effective area: [Formula: see text]with almost no increase in the uncertainty over that at 20 °C.

Cooperative working of bacterial chromosome replication proteins generated by a reconstituted protein expression system

PubMed Central

Fujiwara, Kei; Katayama, Tsutomu; Nomura, Shin-ichiro M.

2013-01-01

Replication of all living cells relies on the multirounds flow of the central dogma. Especially, expression of DNA replication proteins is a key step to circulate the processes of the central dogma. Here we achieved the entire sequential transcription–translation–replication process by autonomous expression of chromosomal DNA replication machineries from a reconstituted transcription–translation system (PURE system). We found that low temperature is essential to express a complex protein, DNA polymerase III, in a single tube using the PURE system. Addition of the 13 genes, encoding initiator, DNA helicase, helicase loader, RNA primase and DNA polymerase III to the PURE system gave rise to a DNA replication system by a coupling manner. An artificial genetic circuit demonstrated that the DNA produced as a result of the replication is able to provide genetic information for proteins, indicating the in vitro central dogma can sequentially undergo two rounds. PMID:23737447

High-level SLP-2 expression and HER-2/neu protein expression are associated with decreased breast cancer patient survival.

PubMed

Cao, Wenfeng; Zhang, Bin; Liu, Yanxue; Li, Hongtao; Zhang, Shiwu; Fu, Li; Niu, Yun; Ning, Liansheng; Cao, Xuchen; Liu, Zhihua; Sun, Baocun

2007-09-01

There is sufficient evidence that human stomatin-like protein 2 (SLP-2) is a novel cancer-related gene. Its protein is overexpressed in many human cancers. SLP-2 can contribute to the promotion of cell growth, cell adhesion, and tumorigenesis in esophageal squamous cell carcinoma and lymph node metastasis in laryngeal squamous cell carcinoma. Immunohistochemical detection of SLP-2, estrogen and progesterone receptors, and HER-2/neu were performed on 263 cases of primary invasive breast cancer with a tissue microarray. Of 263 cases, 138 (52.5%) showed high expression of SLP-2 protein, and 125 (47.5%) showed low or absent expression. In addition, there were significant positive associations between tumor stage and size (P = .020), lymph node metastasis (P < .001), clinical stage (P < .001), distant metastasis (P = .002), and HER-2/neu protein expression (P = .037) and high-level SLP-2 expression. High-level SLP-2 expression was associated with decreased overall survival (P = .011) and was more often found in patients with tumors larger than 20 mm, lymph node metastasis, advanced clinical stage, distant metastasis, and HER-2/neu protein-positive expression. More important, lymph node metastasis, HER-2/neu-positive expression, and high-level SLP-2 expression were associated with significantly decreased survival.

Expression and purification of recombinant proteins in Escherichia coli tagged with the metal-binding protein CusF.

PubMed

Cantu-Bustos, J Enrique; Vargas-Cortez, Teresa; Morones-Ramirez, Jose Ruben; Balderas-Renteria, Isaias; Galbraith, David W; McEvoy, Megan M; Zarate, Xristo

2016-05-01

Production of recombinant proteins in Escherichia coli has been improved considerably through the use of fusion proteins, because they increase protein solubility and facilitate purification via affinity chromatography. In this article, we propose the use of CusF as a new fusion partner for expression and purification of recombinant proteins in E. coli. Using a cell-free protein expression system, based on the E. coli S30 extract, Green Fluorescent Protein (GFP) was expressed with a series of different N-terminal tags, immobilized on self-assembled protein microarrays, and its fluorescence quantified. GFP tagged with CusF showed the highest fluorescence intensity, and this was greater than the intensities from corresponding GFP constructs that contained MBP or GST tags. Analysis of protein production in vivo showed that CusF produces large amounts of soluble protein with low levels of inclusion bodies. Furthermore, fusion proteins can be exported to the cellular periplasm, if CusF contains the signal sequence. Taking advantage of its ability to bind copper ions, recombinant proteins can be purified with readily available IMAC resins charged with this metal ion, producing pure proteins after purification and tag removal. We therefore recommend the use of CusF as a viable alternative to MBP or GST as a fusion protein/affinity tag for the production of soluble recombinant proteins in E. coli. Copyright © 2016 Elsevier Inc. All rights reserved.

Strain Gage Loads Calibration Testing with Airbag Support for the Gulfstream III SubsoniC Research Aircraft Testbed (SCRAT)

NASA Technical Reports Server (NTRS)

Lokos, William A.; Miller, Eric J.; Hudson, Larry D.; Holguin, Andrew C.; Neufeld, David C.; Haraguchi, Ronnie

2015-01-01

This paper describes the design and conduct of the strain-gage load calibration ground test of the SubsoniC Research Aircraft Testbed, Gulfstream III aircraft, and the subsequent data analysis and results. The goal of this effort was to create and validate multi-gage load equations for shear force, bending moment, and torque for two wing measurement stations. For some of the testing the aircraft was supported by three airbags in order to isolate the wing structure from extraneous load inputs through the main landing gear. Thirty-two strain gage bridges were installed on the left wing. Hydraulic loads were applied to the wing lower surface through a total of 16 load zones. Some dead-weight load cases were applied to the upper wing surface using shot bags. Maximum applied loads reached 54,000 lb. Twenty-six load cases were applied with the aircraft resting on its landing gear, and 16 load cases were performed with the aircraft supported by the nose gear and three airbags around the center of gravity. Maximum wing tip deflection reached 17 inches. An assortment of 2, 3, 4, and 5 strain-gage load equations were derived and evaluated against independent check cases. The better load equations had root mean square errors less than 1 percent. Test techniques and lessons learned are discussed.

Adjusted peak-flow frequency estimates for selected streamflow-gaging stations in or near Montana based on data through water year 2011: Chapter D in Montana StreamStats

USGS Publications Warehouse

Sando, Steven K.; Sando, Roy; McCarthy, Peter M.; Dutton, DeAnn M.

2016-04-05

The climatic conditions of the specific time period during which peak-flow data were collected at a given streamflow-gaging station (hereinafter referred to as gaging station) can substantially affect how well the peak-flow frequency (hereinafter referred to as frequency) results represent long-term hydrologic conditions. Differences in the timing of the periods of record can result in substantial inconsistencies in frequency estimates for hydrologically similar gaging stations. Potential for inconsistency increases with decreasing peak-flow record length. The representativeness of the frequency estimates for a short-term gaging station can be adjusted by various methods including weighting the at-site results in association with frequency estimates from regional regression equations (RREs) by using the Weighted Independent Estimates (WIE) program. Also, for gaging stations that cannot be adjusted by using the WIE program because of regulation or drainage areas too large for application of RREs, frequency estimates might be improved by using record extension procedures, including a mixed-station analysis using the maintenance of variance type I (MOVE.1) procedure. The U.S. Geological Survey, in cooperation with the Montana Department of Transportation and the Montana Department of Natural Resources and Conservation, completed a study to provide adjusted frequency estimates for selected gaging stations through water year 2011.The purpose of Chapter D of this Scientific Investigations Report is to present adjusted frequency estimates for 504 selected streamflow-gaging stations in or near Montana based on data through water year 2011. Estimates of peak-flow magnitudes for the 66.7-, 50-, 42.9-, 20-, 10-, 4-, 2-, 1-, 0.5-, and 0.2-percent annual exceedance probabilities are reported. These annual exceedance probabilities correspond to the 1.5-, 2-, 2.33-, 5-, 10-, 25-, 50-, 100-, 200-, and 500-year recurrence intervals, respectively.The at-site frequency estimates were

[Expression of goat IL-18 mature protein in insect/baculovirus and determination of bioactivity of the recombinant protein].

PubMed

Wang, Ting-Ting; Wang, Xi-Hui; Fan, Zhong-Ling; Chen, Jin-Long; Cao, Bing-Lei; Kong, Na; Hu, Jing-Dong; Zhao, Hong-Kun

2011-02-01

To express goat IL-18 in insect/baculovirus and detect the bioactivity of the recombinant protein. The mature goat interleukin-18(gIL-18) gene was cloned into the baculovirus transfer vector pFastBac Dual, and then the resulting eukaryotic expression plasmid pFastBac Dual-gIL18 was transformed into DH10Bac, followed by the identification of Bacmid-gIL18 recombinat plosmid by three antibiotics and blue-white patch. Finally, the recombinant bacmid was transfected into sf9 insect cells by Cellfectin and the transfected cells were harvested at different times. Then the expressed protein was identified by SDS-PAGE, Western blot and bioactivity assay. The recombinant protein recognized and bound to its specific antibody. Bioactivity assay showed that the recombinant protein stimulated the proliferation of lymphocytes and induced IFN-γproduction in spleen lymphocytes. The mature gIL-18 protein has been expressed successfully in insect/baculovirus expression system, and have good immunogenicity and bioactivity. The study paves a way for application of gIL-18 as an immunomodulator or immune adjuvant.

Green fluorescent protein as a reporter of gene expression and protein localization.

PubMed

Kain, S R; Adams, M; Kondepudi, A; Yang, T T; Ward, W W; Kitts, P

1995-10-01

The green fluorescent protein (GFP) from the jellyfish Aequorea victoria is rapidly becoming an important reporter molecule for monitoring gene expression and protein localization in vivo, in situ and in real time. GFP emits bright green light (lambda max = 509 nm) when excited with UV or blue light (lambda max = 395 nm, minor peak at 470 nm). The fluorescence excitation and emission spectra of GFP are similar to those of fluorescein, and the conditions used to visualize this fluorophore are also suitable for GFP. Unlike other bioluminescent reporters, the chromophore in GFP is intrinsic to the primary structure of the protein, and GFP fluorescence does not require a substrate or cofactor. GFP fluorescence is stable, species-independent and can be monitored non-invasively in living cells and, in the case of transparent organisms, whole animals. Here we demonstrate GFP fluorescence in bacterial and mammalian cells and introduce our Living Colors line of GFP reporter vectors, GFP protein and anti-GFP antiserum. The reporter vectors for GFP include a promoterless GFP vector for monitoring the expression of cloned promoters/enhancers in mammalian cells and a series of six vectors for creating fusion protein to either the N or C terminus of GFP.

Heterologous Expression of Membrane Proteins: Choosing the Appropriate Host

PubMed Central

Pochon, Nathalie; Dementin, Sébastien; Hivin, Patrick; Boutigny, Sylvain; Rioux, Jean-Baptiste; Salvi, Daniel; Seigneurin-Berny, Daphné; Richaud, Pierre; Joyard, Jacques; Pignol, David; Sabaty, Monique; Desnos, Thierry; Pebay-Peyroula, Eva; Darrouzet, Elisabeth; Vernet, Thierry; Rolland, Norbert

2011-01-01

Background Membrane proteins are the targets of 50% of drugs, although they only represent 1% of total cellular proteins. The first major bottleneck on the route to their functional and structural characterisation is their overexpression; and simply choosing the right system can involve many months of trial and error. This work is intended as a guide to where to start when faced with heterologous expression of a membrane protein. Methodology/Principal Findings The expression of 20 membrane proteins, both peripheral and integral, in three prokaryotic (E. coli, L. lactis, R. sphaeroides) and three eukaryotic (A. thaliana, N. benthamiana, Sf9 insect cells) hosts was tested. The proteins tested were of various origins (bacteria, plants and mammals), functions (transporters, receptors, enzymes) and topologies (between 0 and 13 transmembrane segments). The Gateway system was used to clone all 20 genes into appropriate vectors for the hosts to be tested. Culture conditions were optimised for each host, and specific strategies were tested, such as the use of Mistic fusions in E. coli. 17 of the 20 proteins were produced at adequate yields for functional and, in some cases, structural studies. We have formulated general recommendations to assist with choosing an appropriate system based on our observations of protein behaviour in the different hosts. Conclusions/Significance Most of the methods presented here can be quite easily implemented in other laboratories. The results highlight certain factors that should be considered when selecting an expression host. The decision aide provided should help both newcomers and old-hands to select the best system for their favourite membrane protein. PMID:22216205

Fe-S Proteins that Regulate Gene Expression

PubMed Central

Mettert, Erin L.; Kiley, Patricia J.

2014-01-01

Iron-sulfur (Fe-S) cluster containing proteins that regulate gene expression are present in most organisms. The innate chemistry of their Fe-S cofactors makes these regulatory proteins ideal for sensing environmental signals, such as gases (e.g. O2 and NO), levels of Fe and Fe-S clusters, reactive oxygen species, and redox cycling compounds, to subsequently mediate an adaptive response. Here we review the recent findings that have provided invaluable insight into the mechanism and function of these highly significant Fe-S regulatory proteins. PMID:25450978

In vitro protein expression: an emerging alternative to cell-based approaches.

PubMed

He, Mingyue

2011-04-30

Protein expression remains a bottleneck in the production of proteins. Owing to several advantages, cell-free translation is emerging as an alternative to cell-based methods for the generation of proteins. Recent advances have led to many novel applications of cell-free systems in biotechnology, proteomics and fundamental biological research. This special issue of New Biotechnology describes recent advances in cell-free protein expression systems and their applications. Copyright © 2010 Elsevier B.V. All rights reserved.

Subcellular localization of transiently expressed fluorescent fusion proteins.

PubMed

Collings, David A

2013-01-01

The recent and massive expansion in plant genomics data has generated a large number of gene sequences for which two seemingly simple questions need to be answered: where do the proteins encoded by these genes localize in cells, and what do they do? One widespread approach to answering the localization question has been to use particle bombardment to transiently express unknown proteins tagged with green fluorescent protein (GFP) or its numerous derivatives. Confocal fluorescence microscopy is then used to monitor the localization of the fluorescent protein as it hitches a ride through the cell. The subcellular localization of the fusion protein, if not immediately apparent, can then be determined by comparison to localizations generated by fluorescent protein fusions to known signalling sequences and proteins, or by direct comparison with fluorescent dyes. This review aims to be a tour guide for researchers wanting to travel this hitch-hiker's path, and for reviewers and readers who wish to understand their travel reports. It will describe some of the technology available for visualizing protein localizations, and some of the experimental approaches for optimizing and confirming localizations generated by particle bombardment in onion epidermal cells, the most commonly used experimental system. As the non-conservation of signal sequences in heterologous expression systems such as onion, and consequent mis-targeting of fusion proteins, is always a potential problem, the epidermal cells of the Argenteum mutant of pea are proposed as a model system.

High-yield, in vitro protein expression using a continuous-exchange, coupled transcription/ translation system.

PubMed

Martin, G A; Kawaguchi, R; Lam, Y; DeGiovanni, A; Fukushima, M; Mutter, W

2001-10-01

The Rapid Translation System (RTS 500) (Roche Molecular Biochemicals) is a high-yield protein expression system that utilizes an enhanced E. coli lysate for an in vitro transcription/translation reaction. In contrast to conventional transcription/translation, this system allows protein expression to continue for more than 24 h. We demonstrated the utility of the RTS 500 by expressing different soluble and active proteins that generally pose problems in cell-based expression systems. We first expressed GFP-lunasin, a fusion protein that, because of its toxicity, has been impossible to produce in whole cells. The second protein we expressed, human interleukin-2 (IL-2), is generally difficult to produce, either as the native molecule or as a GSTfusion protein, in a soluble form in bacteria. Finally, we demonstrated the capacity of the RTS 500 to co-express proteins, by the simultaneous production of GFP and CAT in a single reaction. This new technology appears to be particularly usefulfor the convenient production of preparative amounts (100-900 microg) of proteins that are toxic or insoluble in cell-based systems.

Machine learning in computational biology to accelerate high-throughput protein expression.

PubMed

Sastry, Anand; Monk, Jonathan; Tegel, Hanna; Uhlen, Mathias; Palsson, Bernhard O; Rockberg, Johan; Brunk, Elizabeth

2017-08-15

The Human Protein Atlas (HPA) enables the simultaneous characterization of thousands of proteins across various tissues to pinpoint their spatial location in the human body. This has been achieved through transcriptomics and high-throughput immunohistochemistry-based approaches, where over 40 000 unique human protein fragments have been expressed in E. coli. These datasets enable quantitative tracking of entire cellular proteomes and present new avenues for understanding molecular-level properties influencing expression and solubility. Combining computational biology and machine learning identifies protein properties that hinder the HPA high-throughput antibody production pipeline. We predict protein expression and solubility with accuracies of 70% and 80%, respectively, based on a subset of key properties (aromaticity, hydropathy and isoelectric point). We guide the selection of protein fragments based on these characteristics to optimize high-throughput experimentation. We present the machine learning workflow as a series of IPython notebooks hosted on GitHub (https://github.com/SBRG/Protein_ML). The workflow can be used as a template for analysis of further expression and solubility datasets. ebrunk@ucsd.edu or johanr@biotech.kth.se. Supplementary data are available at Bioinformatics online. © The Author (2017). Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Trends in Streamflow Characteristics at Long-Term Gaging Stations, Hawaii

USGS Publications Warehouse

Oki, Delwyn S.

2004-01-01

The surface-water resources of Hawaii have significant cultural, aesthetic, ecologic, and economic importance. Proper management of the surface-water resources of the State requires an understanding of the long- and short-term variability in streamflow characteristics that may occur. The U.S. Geological Survey maintains a network of stream-gaging stations in Hawaii, including a number of stations with long-term streamflow records that can be used to evaluate long-term trends and short-term variability in flow characteristics. The overall objective of this study is to obtain a better understanding of long-term trends and variations in streamflow on the islands of Hawaii, Maui, Molokai, Oahu, and Kauai, where long-term stream-gaging stations exist. This study includes (1) an analysis of long-term trends in flows (both total flow and estimated base flow) at 16 stream-gaging stations, (2) a description of patterns in trends within the State, and (3) discussion of possible regional factors (including rainfall) that are related to the observed trends and variations. Results of this study indicate the following: 1. From 1913 to 2002 base flows generally decreased in streams for which data are available, and this trend is consistent with the long-term downward trend in annual rainfall over much of the State during that period. 2. Monthly mean base flows generally were above the long-term average from 1913 to the early 1940s and below average after the early 1940s to 2002, and this pattern is consistent with the detected downward trends in base flows from 1913 to 2002. 3. Long-term downward trends in base flows of streams may indicate a reduction in ground-water discharge to streams caused by a long-term decrease in ground-water storage and recharge. 4. From 1973 to 2002, trends in streamflow were spatially variable (up in some streams and down in others) and, with a few exceptions, generally were not statistically significant. 5. Short-term variability in streamflow is

Interfacial Polymerization for Colorimetric Labeling of Protein Expression in Cells

PubMed Central

Lilly, Jacob L.; Sheldon, Phillip R.; Hoversten, Liv J.; Romero, Gabriela; Balasubramaniam, Vivek; Berron, Brad J.

2014-01-01

Determining the location of rare proteins in cells typically requires the use of on-sample amplification. Antibody based recognition and enzymatic amplification is used to produce large amounts of visible label at the site of protein expression, but these techniques suffer from the presence of nonspecific reactivity in the biological sample and from poor spatial control over the label. Polymerization based amplification is a recently developed alternative means of creating an on-sample amplification for fluorescence applications, while not suffering from endogenous labels or loss of signal localization. This manuscript builds upon polymerization based amplification by developing a stable, archivable, and colorimetric mode of amplification termed Polymer Dye Labeling. The basic concept involves an interfacial polymer grown at the site of protein expression and subsequent staining of this polymer with an appropriate dye. The dyes Evans Blue and eosin were initially investigated for colorimetric response in a microarray setting, where both specifically stained polymer films on glass. The process was translated to the staining of protein expression in human dermal fibroblast cells, and Polymer Dye Labeling was specific to regions consistent with desired protein expression. The labeling is stable for over 200 days in ambient conditions and is also compatible with modern mounting medium. PMID:25536421

Interfacial polymerization for colorimetric labeling of protein expression in cells.

PubMed

Lilly, Jacob L; Sheldon, Phillip R; Hoversten, Liv J; Romero, Gabriela; Balasubramaniam, Vivek; Berron, Brad J

2014-01-01

Determining the location of rare proteins in cells typically requires the use of on-sample amplification. Antibody based recognition and enzymatic amplification is used to produce large amounts of visible label at the site of protein expression, but these techniques suffer from the presence of nonspecific reactivity in the biological sample and from poor spatial control over the label. Polymerization based amplification is a recently developed alternative means of creating an on-sample amplification for fluorescence applications, while not suffering from endogenous labels or loss of signal localization. This manuscript builds upon polymerization based amplification by developing a stable, archivable, and colorimetric mode of amplification termed Polymer Dye Labeling. The basic concept involves an interfacial polymer grown at the site of protein expression and subsequent staining of this polymer with an appropriate dye. The dyes Evans Blue and eosin were initially investigated for colorimetric response in a microarray setting, where both specifically stained polymer films on glass. The process was translated to the staining of protein expression in human dermal fibroblast cells, and Polymer Dye Labeling was specific to regions consistent with desired protein expression. The labeling is stable for over 200 days in ambient conditions and is also compatible with modern mounting medium.

The Expression and Significance of Neuronal Iconic Proteins in Podocytes

PubMed Central

Sun, Yu; Zhang, Hongxia; Hu, Ruimin; Sun, Jianyong; Mao, Xing; Zhao, Zhonghua; Chen, Qi; Zhang, Zhigang

2014-01-01

Growing evidence suggests that there are many common cell biological features shared by neurons and podocytes; however, the mechanism of podocyte foot process formation remains unclear. Comparing the mechanisms of process formation between two cell types should provide useful guidance from the progress of neuron research. Studies have shown that some mature proteins of podocytes, such as podocin, nephrin, and synaptopodin, were also expressed in neurons. In this study, using cell biological experiments and immunohistochemical techniques, we showed that some neuronal iconic molecules, such as Neuron-specific enolase, nestin and Neuron-specific nuclear protein, were also expressed in podocytes. We further inhibited the expression of Neuron-specific enolase, nestin, synaptopodin and Ubiquitin carboxy terminal hydrolase-1 by Small interfering RNA in cultured mouse podocytes and observed the significant morphological changes in treated podocytes. When podocytes were treated with Adriamycin, the protein expression of Neuron-specific enolase, nestin, synaptopodin and Ubiquitin carboxy terminal hydrolase-1 decreased over time. Meanwhile, the morphological changes in the podocytes were consistent with results of the Small interfering RNA treatment of these proteins. The data demonstrated that neuronal iconic proteins play important roles in maintaining and regulating the formation and function of podocyte processes. PMID:24699703

p53 AND MDM2 PROTEIN EXPRESSION IN ACTINIC CHEILITIS

PubMed Central

de Freitas, Maria da Conceição Andrade; Ramalho, Luciana Maria Pedreira; Xavier, Flávia Caló Aquino; Moreira, André Luis Gomes; Reis, Sílvia Regina Almeida

2008-01-01

Actinic cheilitis is a potentially malignant lip lesion caused by excessive and prolonged exposure to ultraviolet radiation, which can lead to histomorphological alterations indicative of abnormal cell differentiation. In this pathology, varying degrees of epithelial dysplasia may be found. There are few published studies regarding the p53 and MDM2 proteins in actinic cheilitis. Fifty-eight cases diagnosed with actinic cheilitis were histologically evaluated using Banóczy and Csiba (1976) parameters, and were subjected to immunohistochemical analysis using the streptavidin-biotin method in order to assess p53 and MDM2 protein expression. All studied cases expressed p53 proteins in basal and suprabasal layers. In the basal layer, the nuclei testing positive for p53 were stained intensely, while in the suprabasal layer, cells with slightly stained nuclei were predominant. All cases also tested positive for the MDM2 protein, but with varying degrees of nuclear expression and a predominance of slightly stained cells. A statistically significant correlation between the percentage of p53 and MDM2-positive cells was established, regardless of the degree of epithelial dysplasia. The expression of p53 and MDM2 proteins in actinic cheilitis can be an important indicator in lip carcinogenesis, regardless of the degree of epithelial dysplasia. PMID:19082401

p53 and MDM2 protein expression in actinic cheilitis.

PubMed

de Freitas, Maria da Conceição Andrade; Ramalho, Luciana Maria Pedreira; Xavier, Flávia Caló Aquino; Moreira, André Luis Gomes; Reis, Sílvia Regina Almeida

2008-01-01

Actinic cheilitis is a potentially malignant lip lesion caused by excessive and prolonged exposure to ultraviolet radiation, which can lead to histomorphological alterations indicative of abnormal cell differentiation. In this pathology, varying degrees of epithelial dysplasia may be found. There are few published studies regarding the p53 and MDM2 proteins in actinic cheilitis. Fifty-eight cases diagnosed with actinic cheilitis were histologically evaluated using Banóczy and Csiba (1976) parameters, and were subjected to immunohistochemical analysis using the streptavidin-biotin method in order to assess p53 and MDM2 protein expression. All studied cases expressed p53 proteins in basal and suprabasal layers. In the basal layer, the nuclei testing positive for p53 were stained intensely, while in the suprabasal layer, cells with slightly stained nuclei were predominant. All cases also tested positive for the MDM2 protein, but with varying degrees of nuclear expression and a predominance of slightly stained cells. A statistically significant correlation between the percentage of p53 and MDM2-positive cells was established, regardless of the degree of epithelial dysplasia. The expression of p53 and MDM2 proteins in actinic cheilitis can be an important indicator in lip carcinogenesis, regardless of the degree of epithelial dysplasia.

Development of termination and utilization concepts for flat conductor cables. Volume 2: Utilization of small gage wire round conductor cables

NASA Technical Reports Server (NTRS)

1972-01-01

The practical use of small-gage round wire for electrical wiring in manned air and space vehicle environments is discussed. The investigation consisted on a study of wire construction and candidate wire harness concepts, fabrication of small-gage wire harnesses, and verification of promising configurations by laboratory evaluation. The wire constructions selected for harness fabrication are described. Results of the laboratory evaluation are included.

Differentially Expressed Proteins Associated with Fusarium Head Blight Resistance in Wheat

PubMed Central

Zhang, Xianghui; Fu, Jianming; Hiromasa, Yasuaki; Pan, Hongyu; Bai, Guihua

2013-01-01

Background Fusarium head blight (FHB), mainly caused by Fusarium graminearum, substantially reduces wheat grain yield and quality worldwide. Proteins play important roles in defense against the fungal infection. This study characterized differentially expressed proteins between near-isogenic lines (NILs) contrasting in alleles of Fhb1, a major FHB resistance gene in wheat, to identify proteins underlining FHB resistance of Fhb1. Methods The two-dimensional protein profiles were compared between the Fusarium-inoculated spikes of the two NILs collected 72 h after inoculation. The protein profiles of mock- and Fusarium-inoculated Fhb1+NIL were also compared to identify pathogen-responsive proteins. Results Eight proteins were either induced or upregulated in inoculated Fhb1+NIL when compared with mock-inoculated Fhb1+NIL; nine proteins were either induced or upregulated in the Fusarium-inoculated Fhb1+NIL when compared with Fusarium-inoculated Fhb1−NIL. Proteins that were differentially expressed in the Fhb1+NIL, not in the Fhb1−NIL, after Fusarium inoculation included wheat proteins for defending fungal penetration, photosynthesis, energy metabolism, and detoxification. Conclusions Coordinated expression of the identified proteins resulted in FHB resistance in Fhb1+NIL. The results provide insight into the pathway of Fhb1-mediated FHB resistance. PMID:24376514

[Prokaryotic expression, purification and biological activity analysis of recombinant β-Lactamase protein].

PubMed

Zhou, Xiao-liang; Shi, Pei-ji; Wang, Hao

2011-01-01

To prepare RGD4CβL fusion protein using prokaryotic expression system and evaluate the biological activity of the RGD4CβL. RGD4CβL gene was cloned into pColdII to contruct β-Lactamase prokaryotic expression vector. After transformation, the recombinant vector was induced to express recombinant protein RGD4CβL by IPTG in E.coli BL(DE3). The recombinant protein was purified by Ni-NTA resin under denaturing condition and then dialyzed to renature. The tumor cell targeting ability of the recombinant protein was analyzed by flow cytometric analysis. After cleavage and purification, β-Lactamase moiety showed the expected size of 42 000 on Tricine-SDS-PAGE, and was further confirmed by Western blotting. Based on flow cytometric analysis, the purified protein specially targeted breast cancer cell line MCF-7. This research successfully estiblished a method for prokaryotic expression and purification of β-lactamase. These results suggest the potential use of the protein as an agent for ADEPT.

Developmental expression of Drosophila Wiskott-Aldrich Syndrome family proteins

PubMed Central

Rodriguez-Mesa, Evelyn; Abreu-Blanco, Maria Teresa; Rosales-Nieves, Alicia E.; Parkhurst, Susan M.

2012-01-01

Background Wiskott-Aldrich Syndrome (WASP) family proteins participate in many cellular processes involving rearrangements of the actin cytoskeleton. To the date, four WASP subfamily members have been described in Drosophila: Wash, WASp, SCAR, and Whamy. Wash, WASp, and SCAR are essential during early Drosophila development where they function in orchestrating cytoplasmic events including membrane-cytoskeleton interactions. A mutant for Whamy has not yet been reported. Results We generated monoclonal antibodies that are specific to Drosophila Wash, WASp, SCAR, and Whamy, and use these to describe their spatial and temporal localization patterns. Consistent with the importance of WASP family proteins in flies, we find that Wash, WASp, SCAR, and Whamy are dynamically expressed throughout oogenesis and embryogenesis. For example, we find that Wash accumulates at the oocyte cortex. WASp is highly expressed in the PNS, while SCAR is the most abundantly expressed in the CNS. Whamy exhibits an asymmetric subcellular localization that overlaps with mitochondria and is highly expressed in muscle. Conclusion All four WASP family members show specific expression patterns, some of which reflect their previously known roles and others revealing new potential functions. The monoclonal antibodies developed offer valuable new tools to investigate how WASP family proteins regulate actin cytoskeleton dynamics. PMID:22275148

Establishment of stable cell line for inducing KAP1 protein expression.

PubMed

Liu, Xiaoyan; Khan, Md Asaduzzaman; Cheng, Jingliang; Wei, Chunli; Zhang, Lianmei; Fu, Junjiang

2015-06-01

Generation of the stable cell lines is a highly efficient tool in functional studies of certain genes or proteins, where the particular genes or proteins are inducibly expressed. The KRAB-associated protein-1 (KAP1) is an important transcription regulatory protein, which is investigated in several molecular biological studies. In this study, we have aimed to generate a stable cell line for inducing KAP1 expression. The recombinant plasmid pcDNA5/FRT/TO-KAP1 was constructed at first, which was then transfected into Flp-In™T-REx™-HEK293 cells to establish an inducible pcDNA5/FRT/TO-KAP1-HEK293 cell line. The Western blot analysis showed that the protein level of KAP1 is over-expressed in the established stable cell line by doxycycline induction, both dose and time dependently. Thus we have successfully established stable pcDNA5/FRT/TO-KAP1-HEK293 cell line, which can express KAP1 inducibly. This inducible cell line might be very useful for KAP1 functional studies.

Unintended changes in protein expression revealed by proteomic analysis of seeds from transgenic pea expressing a bean alpha-amylase inhibitor gene.

PubMed

Chen, Hancai; Bodulovic, Greg; Hall, Prudence J; Moore, Andy; Higgins, Thomas J V; Djordjevic, Michael A; Rolfe, Barry G

2009-09-01

Seeds of genetically modified (GM) peas (Pisum sativum L.) expressing the gene for alpha-amylase inhibitor-1 (alphaAI1) from the common bean (Phaseolus vulgaris L. cv. Tendergreen) exhibit resistance to the pea weevil (Bruchus pisorum). A proteomic analysis was carried out to compare seeds from GM pea lines expressing the bean alphaAI1 protein and the corresponding alphaAI1-free segregating lines and non-GM parental line to identify unintended alterations to the proteome of GM peas due to the introduction of the gene for alphaAI1. Proteomic analysis showed that in addition to the presence of alphaAI1, 33 other proteins were differentially accumulated in the alphaAI1-expressing GM lines compared with their non-GM parental line and these were grouped into five expression classes. Among these 33 proteins, only three were found to be associated with the expression of alphaAI1 in the GM pea lines. The accumulation of the remaining 30 proteins appears to be associated with Agrobacterium-mediated transformation events. Sixteen proteins were identified after MALDI-TOF-TOF analysis. About 56% of the identified proteins with altered accumulation in the GM pea were storage proteins including legumin, vicilin or convicilin, phaseolin, cupin and valosin-containing protein. Two proteins were uniquely expressed in the alphaAI1-expressing GM lines and one new protein was present in both the alphaAI1-expressing GM lines and their alphaAI1-free segregating lines, suggesting that both transgenesis and transformation events led to demonstrable changes in the proteomes of the GM lines tested.

Variation in Protein Intake Induces Variation in Spider Silk Expression

PubMed Central

Blamires, Sean J.; Wu, Chun-Lin; Tso, I-Min

2012-01-01

Background It is energetically expensive to synthesize certain amino acids. The proteins (spidroins) of spider major ampullate (MA) silk, MaSp1 and MaSp2, differ in amino acid composition. Glutamine and proline are prevalent in MaSp2 and are expensive to synthesize. Since most orb web spiders express high proline silk they might preferentially attain the amino acids needed for silk from food and shift toward expressing more MaSp1 in their MA silk when starved. Methodology/Principal Findings We fed three spiders; Argiope aetherea, Cyrtophora moluccensis and Leucauge blanda, high protein, low protein or no protein solutions. A. aetherea and L. blanda MA silks are high in proline, while C. moluccesnsis MA silks are low in proline. After 10 days of feeding we determined the amino acid compositions and mechanical properties of each species' MA silk and compared them between species and treatments with pre-treatment samples, accounting for ancestry. We found that the proline and glutamine of A. aetherea and L. blanda silks were affected by protein intake; significantly decreasing under the low and no protein intake treatments. Glutmaine composition in C. moluccensis silk was likewise affected by protein intake. However, the composition of proline in their MA silk was not significantly affected by protein intake. Conclusions Our results suggest that protein limitation induces a shift toward different silk proteins with lower glutamine and/or proline content. Contradictions to the MaSp model lie in the findings that C. moluccensis MA silks did not experience a significant reduction in proline and A. aetherea did not experience a significant reduction in serine on low/no protein. The mechanical properties of the silks could not be explained by a MaSp1 expressional shift. Factors other than MaSp expression, such as the expression of spidroin-like orthologues, may impact on silk amino acid composition and spinning and glandular processes may impact mechanics. PMID:22363691

[Phineas Gage, 'Tan' and the importance of case reports].

PubMed

Rosselli, D

The original descriptions of the frontal lobe injury of Phineas Gage (1848) and the slowly growing tumor of 'Tan', Broca's famous patient (1861), are examples of how a simple case report can teach important lessons, some of them still discussed a century and a half later. In this article, the original sources of both of these seminal cases, in Boston and Paris, have been reviewed and are briefly summarized. The lessons learned from them in the effort to localize brain functions are explained and set in the context of modern evidence-based medicine.

RRE-dependent HIV-1 Env RNA effects on Gag protein expression, assembly and release

DOE Office of Scientific and Technical Information (OSTI.GOV)

López, Claudia S., E-mail: lopezcl@ohsu.edu; Sloan, Rachel; Cylinder, Isabel

The HIV-1 Gag proteins are translated from the full-length HIV-1 viral RNA (vRNA), whereas the envelope (Env) protein is translated from incompletely spliced Env mRNAs. Nuclear export of vRNAs and Env mRNAs is mediated by the Rev accessory protein which binds to the rev-responsive element (RRE) present on these RNAs. Evidence has shown there is a direct or indirect interaction between the Gag protein, and the cytoplasmic tail (CT) of the Env protein. Our current work shows that env gene expression impacts HIV-1 Gag expression and function in two ways. At the protein level, full-length Env expression altered Gag proteinmore » expression, while Env CT-deletion proteins did not. At the RNA level, RRE-containing Env mRNA expression reduced Gag expression, processing, and virus particle release from cells. Our results support models in which Gag is influenced by the Env CT, and Env mRNAs compete with vRNAs for nuclear export. - Highlights: • At the protein level, full-length HIV-1 Env alters Gag protein expression. • HIV-1 Env RNA expression reduces Gag levels and virus release. • Env RNA effects on Gag are dependent on the RRE. • RRE-containing Env RNAs compete with vRNAs for nuclear export.« less

Hippocampal expression of a virus-derived protein impairs memory in mice.

PubMed

Bétourné, Alexandre; Szelechowski, Marion; Thouard, Anne; Abrial, Erika; Jean, Arnaud; Zaidi, Falek; Foret, Charlotte; Bonnaud, Emilie M; Charlier, Caroline M; Suberbielle, Elsa; Malnou, Cécile E; Granon, Sylvie; Rampon, Claire; Gonzalez-Dunia, Daniel

2018-02-13

The analysis of the biology of neurotropic viruses, notably of their interference with cellular signaling, provides a useful tool to get further insight into the role of specific pathways in the control of behavioral functions. Here, we exploited the natural property of a viral protein identified as a major effector of behavioral disorders during infection. We used the phosphoprotein (P) of Borna disease virus, which acts as a decoy substrate for protein kinase C (PKC) when expressed in neurons and disrupts synaptic plasticity. By a lentiviral-based strategy, we directed the singled-out expression of P in the dentate gyrus of the hippocampus and we examined its impact on mouse behavior. Mice expressing the P protein displayed increased anxiety and impaired long-term memory in contextual and spatial memory tasks. Interestingly, these effects were dependent on P protein phosphorylation by PKC, as expression of a mutant form of P devoid of its PKC phosphorylation sites had no effect on these behaviors. We also revealed features of behavioral impairment induced by P protein expression but that were independent of its phosphorylation by PKC. Altogether, our findings provide insight into the behavioral correlates of viral infection, as well as into the impact of virus-mediated alterations of the PKC pathway on behavioral functions.

BAX protein expression and clinical outcome in epithelial ovarian cancer.

PubMed

Tai, Y T; Lee, S; Niloff, E; Weisman, C; Strobel, T; Cannistra, S A

1998-08-01

Expression of the pro-apoptotic protein BAX sensitizes ovarian cancer cell lines to paclitaxel in vitro by enhancing the pathway of programmed cell death. The present study was performed to determine the relationship between BAX expression and clinical outcome in 45 patients with newly diagnosed ovarian cancer. BAX protein expression was analyzed by immunohistochemistry, and its relationship with clinical outcome was determined. Assessment of BAX mRNA transcript levels and mutational analysis of the BAX coding region were also performed. BAX protein was expressed at high levels (defined as > or = 50% of tumor cells positive) in tumor tissue from 60% of newly diagnosed patients. All patients whose tumors expressed high levels of BAX achieved a complete response (CR) to first-line chemotherapy that contained paclitaxel plus a platinum analogue, compared with 57% of patients in the low-BAX group (P = .036). After a median follow-up of 1.9 years, the median disease-free survival (DFS) of patients in the high-BAX group has not been reached, compared with a median DFS of 1.1 years for low-BAX expressors (P = .0061). BAX retained independent prognostic significance in multivariate analysis when corrected for stage and histology. BAX mRNA transcripts were easily detected in samples with low BAX protein expression, and no BAX mutations were identified. The correlation between high BAX levels and improved clinical outcome suggests that an intact apoptotic pathway is an important determinant of chemoresponsiveness in ovarian cancer patients who receive paclitaxel.

Optimization of laser welding thin-gage galvanized steel via response surface methodology

NASA Astrophysics Data System (ADS)

Zhao, Yangyang; Zhang, Yansong; Hu, Wei; Lai, Xinmin

2012-09-01

The increasing demand of light weight and durability makes thin-gage galvanized steels (<0.6 mm) attractive for future automotive applications. Laser welding, well known for its deep penetration, high speed and small heat affected zone, provides a potential solution for welding thin-gage galvanized steels in automotive industry. In this study, the effect of the laser welding parameters (i.e. laser power, welding speed, gap and focal position) on the weld bead geometry (i.e. weld depth, weld width and surface concave) of 0.4 mm-thick galvanized SAE1004 steel in a lap joint configuration has been investigated by experiments. The process windows of the concerned process parameters were therefore determined. Then, response surface methodology (RSM) was used to develop models to predict the relationship between the processing parameters and the laser weld bead profile and identify the correct and optimal combination of the laser welding input variables to obtain superior weld joint. Under the optimal welding parameters, defect-free weld were produced, and the average aspect ratio increased about 30%, from 0.62 to 0.83.

Exploratory Calibration of Adjustable-Protrusion Surface-Obstacle (APSO) Skin Friction Vector Gage

NASA Technical Reports Server (NTRS)

Hakkinen, Raimo J.; Neubauer, Jeremy S.; Hamory, Philip J.; Bui, Trong T.; Noffz, Gregory K.; Young, Ron (Technical Monitor)

2003-01-01

The design of an adjustable-protrusion surface-obstacle (APSO) skin friction vector gage is presented. Results from exploratory calibrations conducted in laminar and turbulent boundary layers at the Washington University Low-Speed Wind Tunnel and for turbulent boundary layers at speeds up to Mach 2 on the ceiling of the NASA Glenn Research Center 8- X 6-ft Supersonic Wind Tunnel are also discussed. The adjustable-height gage was designed to yield both the magnitude and direction of the surface shear stress vector and to measure the local static pressure distribution. Results from the NASA test show good correlation for subsonic and low supersonic conditions covering several orders of magnitude in terms of the adopted similarity variables. Recommendations for future work in this area consist of identifying the physical parameters responsible for the disagreement between the university and NASA data sets, developing a compressibility correction specific to the APSO geometry, and examining the effect that static pressure distribution and skewed boundary layers have on the results from the APSO.

Protein Expression Profile using Two-Dimensional Gel Analysis in Squamous Cervical Cancer Patients

PubMed Central

Bae, Su-Mi; Min, Hyun-Jin; Ding, Guo Hua; Kwak, Sun-Young; Cho, Young-Lae; Nam, Kye-Hyun; Park, Choong Hak; Kim, Yong-Wan; Kim, Chong-Kook; Han, Byoung-Don; Lee, Young-Joo; Kim, Do Kang

2006-01-01

Purpose Screening in cervical cancer is now progressing to discover candidate genes and proteins that may serve as biological markers and that play a role in tumor progression. We examined the protein expression patterns of the squamous cell carcinoma (SCC) tissues from Korean women with using two- dimensional polyacrylamide gel electrophoresis (2-DE) and matrix assisted laser desorption/ionization-time of flight (MALDI- TOF) mass spectrometer. Materials and Methods Normal cervix and SCC tissues were solubilized and 2-DE was performed using pH 3~10 linear IPG strips of 17 cm length. The protein expression was evaluated using PDQuest 2-D software™. The differentially expressed protein spots were identified with a MALDI-TOF mass spectrometer, and the peptide mass spectra identifications were performed using the Mascot program and by searching the Swiss-prot or NCBInr databases. Results A total of 35 proteins were detected in SCC. 17 proteins were up-regulated and 18 proteins weredown-regulated. Among the proteins that were identified, 12 proteins (pigment epithelium derived factor, annexin A2 and A5, keratin 19 and 20, heat shock protein 27, smooth muscle protein 22 alpha, α-enolase, squamous cell carcinoma antigen 1 and 2, glutathione S-transferase and apolipoprotein a1) were protein previously known to be involved in tumor, and 21 proteins were newly identified in this study. Conclusion 2-DE offers the total protein expression profiles of SCC tissues; further characterization of these differentially expressed proteins will give a chance to identify the badly needed tumor-specific diagnostic markers for SCC. PMID:19771267

Flight loads measurements obtained from calibrated strain-gage bridges mounted externally on the skin of a low-aspect-ratio wing

NASA Technical Reports Server (NTRS)

Eckstrom, C. V.

1976-01-01

Flight-test measurements of wingloads (shear, bending moment, and torque) were obtained by means of strain-gage bridges mounted on the exterior surface of a low-aspect-ratio, thin, swept wing which had a structural skin, full-depth honeycomb core, sandwich construction. Details concerning the strain-gage bridges, the calibration procedures used, and the flight-test results are presented along with some pressure measurements and theoretical calculations for comparison purposes.

Combined protein construct and synthetic gene engineering for heterologous protein expression and crystallization using Gene Composer

DOE Office of Scientific and Technical Information (OSTI.GOV)

Raymond, Amy; Lovell, Scott; Lorimer, Don

2009-12-01

With the goal of improving yield and success rates of heterologous protein production for structural studies we have developed the database and algorithm software package Gene Composer. This freely available electronic tool facilitates the information-rich design of protein constructs and their engineered synthetic gene sequences, as detailed in the accompanying manuscript. In this report, we compare heterologous protein expression levels from native sequences to that of codon engineered synthetic gene constructs designed by Gene Composer. A test set of proteins including a human kinase (P38{alpha}), viral polymerase (HCV NS5B), and bacterial structural protein (FtsZ) were expressed in both E. colimore » and a cell-free wheat germ translation system. We also compare the protein expression levels in E. coli for a set of 11 different proteins with greatly varied G:C content and codon bias. The results consistently demonstrate that protein yields from codon engineered Gene Composer designs are as good as or better than those achieved from the synonymous native genes. Moreover, structure guided N- and C-terminal deletion constructs designed with the aid of Gene Composer can lead to greater success in gene to structure work as exemplified by the X-ray crystallographic structure determination of FtsZ from Bacillus subtilis. These results validate the Gene Composer algorithms, and suggest that using a combination of synthetic gene and protein construct engineering tools can improve the economics of gene to structure research.« less

In planta expression of HIV-1 p24 protein using an RNA plant virus-based expression vector.

PubMed

Zhang, G; Leung, C; Murdin, L; Rovinski, B; White, K A

2000-02-01

Plant viruses show significant potential as expression vectors for the production of foreign proteins (e.g., antigens) in plants. The HIV-1 p24 nucleocapsid protein is an important early marker of HIV infection and has been used as an antigen in the development of HIV vaccines. Toward developing a plant-based expression system for the production of p24, we have investigated the use of a (positive)-strand RNA plant virus, tomato bushy stunt virus (TBSV), as an expression vector. The HIV p24 open reading frame (ORF) was introduced into a cloned cDNA copy of the TBSV genome as an in-frame fusion with a 5'-terminal portion of the TBSV coat protein ORF. In vitro-generated RNA transcripts corresponding to the engineered virus vector were infectious when inoculated into plant protoplasts; Northern and Western blot analyses verified the accumulation of a predicted p24-encoding viral subgenomic mRNA and the production of p24 fusion product. Whole-plant infections with the viral vector led to the accumulation of p24 fusion protein in inoculated leaves, which cross-reacted with p24-specific antibodies, thus confirming the maintenance of key antigenic determinants. This study is the first to demonstrate that TBSV can be engineered to express a complete foreign protein of clinical importance. Strategies for optimizing protein yield from this viral vector are discussed.

The Center for Optimized Structural Studies (COSS) platform for automation in cloning, expression, and purification of single proteins and protein-protein complexes.

PubMed

Mlynek, Georg; Lehner, Anita; Neuhold, Jana; Leeb, Sarah; Kostan, Julius; Charnagalov, Alexej; Stolt-Bergner, Peggy; Djinović-Carugo, Kristina; Pinotsis, Nikos

2014-06-01

Expression in Escherichia coli represents the simplest and most cost effective means for the production of recombinant proteins. This is a routine task in structural biology and biochemistry where milligrams of the target protein are required in high purity and monodispersity. To achieve these criteria, the user often needs to screen several constructs in different expression and purification conditions in parallel. We describe a pipeline, implemented in the Center for Optimized Structural Studies, that enables the systematic screening of expression and purification conditions for recombinant proteins and relies on a series of logical decisions. We first use bioinformatics tools to design a series of protein fragments, which we clone in parallel, and subsequently screen in small scale for optimal expression and purification conditions. Based on a scoring system that assesses soluble expression, we then select the top ranking targets for large-scale purification. In the establishment of our pipeline, emphasis was put on streamlining the processes such that it can be easily but not necessarily automatized. In a typical run of about 2 weeks, we are able to prepare and perform small-scale expression screens for 20-100 different constructs followed by large-scale purification of at least 4-6 proteins. The major advantage of our approach is its flexibility, which allows for easy adoption, either partially or entirely, by any average hypothesis driven laboratory in a manual or robot-assisted manner.

Comparison of Heat Flux Gages for High Enthalpy Flows - NASA Ames and IRS

NASA Technical Reports Server (NTRS)

Loehle, Stefan; Nawaz, Anuscheh; Herdrich, Georg; Fasoulas, Stefanos; Martinez, Edward; Raiche, George

2016-01-01

This article is a companion to a paper on heat flux measurements as initiated under a Space Act Agreement in 2011. The current focus of this collaboration between the Institute of Space Systems (IRS) of the University of Stuttgart and NASA Ames Research Center is the comparison and refinement of diagnostic measurements. A first experimental campaign to test different heat flux gages in the NASA Interaction Heating Facility (IHF) and the Plasmawindkanaele (PWK) at IRS was established. This paper focuses on the results of the measurements conducted at IRS. The tested gages included a at face and hemispherical probe head, a 4" hemispherical slug calorimeter, a null-point calorimeter from Ames and a null-point calorimeter developed for this purpose at IRS. The Ames null-point calorimeter was unfortunately defective upon arrival. The measured heat fluxes agree fairly well with each other. The reason for discrepancies can be attributed to signal-to-noise levels and the probe geometry.

Preferential expression and immunogenicity of HIV-1 Tat fusion protein expressed in tomato plant.

PubMed

Cueno, Marni E; Hibi, Yurina; Karamatsu, Katsuo; Yasutomi, Yasuhiro; Imai, Kenichi; Laurena, Antonio C; Okamoto, Takashi

2010-10-01

HIV-1 Tat plays a major role in viral replication and is essential for AIDS development making it an ideal vaccine target providing that both humoral and cellular immune responses are induced. Plant-based antigen production, due to its cheaper cost, appears ideal for vaccine production. In this study, we created a plant-optimized tat and mutant (Cys30Ala/Lys41Ala) tat (mtat) gene and ligated each into a pBI121 expression vector with a stop codon and a gusA gene positioned immediately downstream. The vector construct was bombarded into tomato leaf calli and allowed to develop. We thus generated recombinant tomato plants preferentially expressing a Tat-GUS fusion protein over a Tat-only protein. In addition, plants bombarded with either tat or mtat genes showed no phenotypic difference and produced 2-4 microg Tat-GUS fusion protein per milligram soluble plant protein. Furthermore, tomato extracts intradermally inoculated into mice were found to induce a humoral and, most importantly, cellular immunity.

Using Green and Red Fluorescent Proteins to Teach Protein Expression, Purification, and Crystallization

ERIC Educational Resources Information Center

Wu, Yifeng; Zhou, Yangbin; Song, Jiaping; Hu, Xiaojian; Ding, Yu; Zhang, Zhihong

2008-01-01

We have designed a laboratory curriculum using the green and red fluorescent proteins (GFP and RFP) to visualize the cloning, expression, chromatography purification, crystallization, and protease-cleavage experiments of protein science. The EGFP and DsRed monomer (mDsRed)-coding sequences were amplified by PCR and cloned into pMAL (MBP-EGFP) or…

Protein half-life determines expression of proteostatic networks in podocyte differentiation.

PubMed

Schroeter, Christina B; Koehler, Sybille; Kann, Martin; Schermer, Bernhard; Benzing, Thomas; Brinkkoetter, Paul T; Rinschen, Markus M

2018-04-25

Podocytes are highly specialized, epithelial, postmitotic cells, which maintain the renal filtration barrier. When adapting to considerable metabolic and mechanical stress, podocytes need to accurately maintain their proteome. Immortalized podocyte cell lines are a widely used model for studying podocyte biology in health and disease in vitro. In this study, we performed a comprehensive proteomic analysis of the cultured human podocyte proteome in both proliferative and differentiated conditions at a depth of >7000 proteins. Similar to mouse podocytes, human podocyte differentiation involved a shift in proteostasis: undifferentiated podocytes have high expression of proteasomal proteins, whereas differentiated podocytes have high expression of lysosomal proteins. Additional analyses with pulsed stable-isotope labeling by amino acids in cell culture and protein degradation assays determined protein dynamics and half-lives. These studies unraveled a globally increased stability of proteins in differentiated podocytes. Mitochondrial, cytoskeletal, and membrane proteins were stabilized, particularly in differentiated podocytes. Importantly, protein half-lives strongly contributed to protein abundance in each state. These data suggest that regulation of protein turnover of particular cellular functions determines podocyte differentiation, a paradigm involving mitophagy and, potentially, of importance in conditions of increased podocyte stress and damage.-Schroeter, C. B., Koehler, S., Kann, M., Schermer, B., Benzing, T., Brinkkoetter, P. T., Rinschen, M. M. Protein half-life determines expression of proteostatic networks in podocyte differentiation.

An inducible expression system for high-level expression of recombinant proteins in slow growing mycobacteria.

PubMed

Leotta, Lisa; Spratt, Joanne M; Kong, Carlyn U; Triccas, James A

2015-09-01

A novel protein expression vector utilising the inducible hspX promoter of Mycobacterium tuberculosis was constructed and evaluated in this study. High-level induction of three mycobacterial antigens, comprising up to 9% of bacterial sonicate, was demonstrated in recombinant Mycobacterium bovis BCG when grown under low-oxygen tension, which serves to enhance hspX promoter activity. Recombinant proteins were efficiently purified from bacterial lysates in a soluble form by virtue of a C-terminal 6-histidine tag. Purification of the immunodominant M. tuberculosis Ag85B antigen using this system resulted in a recombinant protein that stimulated significant IFN-γ release from Ag85B-reactive T cells generated after vaccination of mice with an Ag85B-expressing vaccine. Further, the M. tuberculosis L-alanine dehydrogenase (Ald) protein purified from recombinant BCG displayed strong enzymatic activity in recombinant form. This study demonstrated that high levels of native-like recombinant mycobacterial proteins can be produced in mycobacterial hosts, and this may aid the analysis of mycobacterial protein function and the development of new treatments. Copyright © 2015 Elsevier B.V. All rights reserved.

Expression of c-Fes protein isoforms correlates with differentiation in myeloid leukemias.

PubMed

Carlson, Anne; Berkowitz, Jeanne McAdara; Browning, Damaris; Slamon, Dennis J; Gasson, Judith C; Yates, Karen E

2005-05-01

The cellular fes gene encodes a 93-kilodalton protein-tyrosine kinase (p93) that is expressed in both normal and neoplastic myeloid cells. Increased c-Fes expression is associated with differentiation in normal myeloid cells and cell lines. Our hypothesis was that primary leukemia cells would show a similar pattern of increased expression in more differentiated cells. Therefore, we compared c-Fes expression in cells with an undifferentiated, blast phenotype (acute myelogenous leukemia--AML) to cells with a differentiated phenotype (chronic myelogenous leukemia--CML). Instead of differences in p93 expression levels, we found complex patterns of c-Fes immunoreactive proteins that corresponded with differentiation in normal and leukemic myeloid cells. The "blast" pattern consisted of c-Fes immunoreactive proteins p93, p74, and p70; the "differentiated" pattern showed two additional c-Fes immunoreactive proteins, p67 and p62. Using mRNA from mouse and human cell lines, we found deletion of one or more exons in the c-fes mRNA. Those deletions predicted truncation of conserved domains (CDC15/FCH and SH2) involved in protein-protein interactions. No deletions were found, however, within the kinase domain. We infer that alternative splicing generates a family of c-Fes proteins. This may be a mechanism to direct the c-Fes kinase domain to different subcellular locations and/or substrates at specific stages of myeloid cell differentiation.

Expression of uncoupling protein 3 is upregulated in skeletal muscle during sepsis.

PubMed

Sun, Xiaoyan; Wray, Curtis; Tian, Xintian; Hasselgren, Per-Olof; Lu, James

2003-09-01

Uncoupling protein 3 (UCP3) is a member of the mitochondrial transporter superfamily that is expressed primarily in skeletal muscle. UCP3 is upregulated in various conditions characterized by skeletal muscle atrophy, including hyperthyroidism, fasting, denervation, diabetes, cancer, lipopolysaccharide (LPS), and treatment with glucocorticoids (GCs). The influence of sepsis, another condition characterized by muscle cachexia, on UCP3 expression and activity is not known. We examined UCP3 gene and protein expression in skeletal muscles from rats after cecal ligation and puncture and from sham-operated control rats. Sepsis resulted in a two- to threefold increase in both mRNA and protein levels of UCP3 in skeletal muscle. Treatment of rats with the glucocorticoid receptor antagonist RU-38486 prevented the sepsis-induced increase in gene and protein expression of UCP3. The UCP3 mRNA and protein levels were increased 2.4- to 3.6-fold when incubated muscles from normal rats were treated with dexamethasone (DEX) and/or free fatty acids (FFA) ex vivo. In addition, UCP3 mRNA and protein levels were significantly increased in normal rat muscles in vivo with treatment of either DEX or FFA. The results suggest that sepsis upregulates the gene and protein expression of UCP3 in skeletal muscle, which may at least in part be mediated by GCs and FFA.

Functional expression of the taste-modifying protein, miraculin, in transgenic lettuce.

PubMed

Sun, Hyeon-Jin; Cui, Min-Long; Ma, Biao; Ezura, Hiroshi

2006-01-23

Taste-modifying proteins are a natural alternative to artificial sweeteners and flavor enhancers and have been used in some cultures for centuries. The taste-modifying protein, miraculin, has the unusual property of being able to modify a sour taste into a sweet taste. Here, we report the use of a plant expression system for the production of miraculin. A synthetic gene encoding miraculin was placed under the control of constitutive promoters and transferred to lettuce. Expression of this gene in transgenic lettuce resulted in the accumulation of significant amounts of miraculin protein in the leaves. The miraculin expressed in transgenic lettuce possessed sweetness-inducing activity. These results demonstrate that the production of miraculin in edible plants can be a good alternative strategy to enhance the availability of this protein.

Genome-Wide Identification and Expression of Xenopus F-Box Family of Proteins.

PubMed

Saritas-Yildirim, Banu; Pliner, Hannah A; Ochoa, Angelica; Silva, Elena M

2015-01-01

Protein degradation via the multistep ubiquitin/26S proteasome pathway is a rapid way to alter the protein profile and drive cell processes and developmental changes. Many key regulators of embryonic development are targeted for degradation by E3 ubiquitin ligases. The most studied family of E3 ubiquitin ligases is the SCF ubiquitin ligases, which use F-box adaptor proteins to recognize and recruit target proteins. Here, we used a bioinformatics screen and phylogenetic analysis to identify and annotate the family of F-box proteins in the Xenopus tropicalis genome. To shed light on the function of the F-box proteins, we analyzed expression of F-box genes during early stages of Xenopus development. Many F-box genes are broadly expressed with expression domains localized to diverse tissues including brain, spinal cord, eye, neural crest derivatives, somites, kidneys, and heart. All together, our genome-wide identification and expression profiling of the Xenopus F-box family of proteins provide a foundation for future research aimed to identify the precise role of F-box dependent E3 ubiquitin ligases and their targets in the regulatory circuits of development.

Basin characteristics, history of stream gaging, and statistical summary of selected streamflow records for the Rapid Creek basin, western South Dakota

USGS Publications Warehouse

Driscoll, Daniel G.; Zogorski, John S.

1990-01-01

The report presents a summary of basin characteristics affecting streamflow, a history of the U.S. Geological Survey 's stream-gaging program, and a compilation of discharge records and statistical summaries for selected sites within the Rapid Creek basin. It is the first in a series which will investigate surface-water/groundwater relations along Rapid Creek. The summary of basin characteristics includes descriptions of the geology and hydrogeology, physiography and climate, land use and vegetation, reservoirs, and water use within the basin. A recounting of the U.S. Geological Survey 's stream-gaging program and a tabulation of historic stream-gaging stations within the basin are furnished. A compilation of monthly and annual mean discharge values for nine currently operated, long-term, continuous-record, streamflow-gaging stations on Rapid Creek is presented. The statistical summary for each site includes summary statistics on monthly and annual mean values, correlation matrix for monthly values, serial correlation for 1 year lag for monthly values, percentile rankings for monthly and annual mean values, low and high value tables, duration curves, and peak-discharge tables. Records of monthend contents for two reservoirs within the basin also are presented. (USGS)

Expressing the human proteome for affinity proteomics: optimising expression of soluble protein domains and in vivo biotinylation.

PubMed

Keates, Tracy; Cooper, Christopher D O; Savitsky, Pavel; Allerston, Charles K; Phillips, Claire; Hammarström, Martin; Daga, Neha; Berridge, Georgina; Mahajan, Pravin; Burgess-Brown, Nicola A; Müller, Susanne; Gräslund, Susanne; Gileadi, Opher

2012-06-15

The generation of affinity reagents to large numbers of human proteins depends on the ability to express the target proteins as high-quality antigens. The Structural Genomics Consortium (SGC) focuses on the production and structure determination of human proteins. In a 7-year period, the SGC has deposited crystal structures of >800 human protein domains, and has additionally expressed and purified a similar number of protein domains that have not yet been crystallised. The targets include a diversity of protein domains, with an attempt to provide high coverage of protein families. The family approach provides an excellent basis for characterising the selectivity of affinity reagents. We present a summary of the approaches used to generate purified human proteins or protein domains, a test case demonstrating the ability to rapidly generate new proteins, and an optimisation study on the modification of >70 proteins by biotinylation in vivo. These results provide a unique synergy between large-scale structural projects and the recent efforts to produce a wide coverage of affinity reagents to the human proteome. Copyright © 2011 Elsevier B.V. All rights reserved.

Expressing the human proteome for affinity proteomics: optimising expression of soluble protein domains and in vivo biotinylation

PubMed Central

Keates, Tracy; Cooper, Christopher D.O.; Savitsky, Pavel; Allerston, Charles K.; Phillips, Claire; Hammarström, Martin; Daga, Neha; Berridge, Georgina; Mahajan, Pravin; Burgess-Brown, Nicola A.; Müller, Susanne; Gräslund, Susanne; Gileadi, Opher

2012-01-01

The generation of affinity reagents to large numbers of human proteins depends on the ability to express the target proteins as high-quality antigens. The Structural Genomics Consortium (SGC) focuses on the production and structure determination of human proteins. In a 7-year period, the SGC has deposited crystal structures of >800 human protein domains, and has additionally expressed and purified a similar number of protein domains that have not yet been crystallised. The targets include a diversity of protein domains, with an attempt to provide high coverage of protein families. The family approach provides an excellent basis for characterising the selectivity of affinity reagents. We present a summary of the approaches used to generate purified human proteins or protein domains, a test case demonstrating the ability to rapidly generate new proteins, and an optimisation study on the modification of >70 proteins by biotinylation in vivo. These results provide a unique synergy between large-scale structural projects and the recent efforts to produce a wide coverage of affinity reagents to the human proteome. PMID:22027370

Exercise alters myostatin protein expression in sedentary and exercised streptozotocin-diabetic rats.

PubMed

Bassi, Daniela; Bueno, Patricia de Godoy; Nonaka, Keico Okino; Selistre-Araujo, Heloisa Sobreiro; Leal, Angela Merice de Oliveira

2015-04-01

The aim of this study was to analyze the effect of exercise on the pattern of muscle myostatin (MSTN) protein expression in two important metabolic disorders, i.e., obesity and diabetes mellitus. MSTN, is a negative regulator of skeletal muscle mass. We evaluated the effect of exercise on MSTN protein expression in diabetes mellitus and high fat diet-induced obesity. MSTN protein expression in gastrocnemius muscle was analyzed by Western Blot. P < 0.05 was assumed. Exercise induced a significant decrease in glycemia in both diabetic and obese animals. The expression of precursor and processed protein forms of MSTN and the weight of gastrocnemius muscle did not vary in sedentary or exercised obese animals. Diabetes reduced gastrocnemius muscle weight in sedentary animals. However, gastrocnemius muscle weight increased in diabetic exercised animals. Both the precursor and processed forms of muscle MSTN protein were significantly higher in sedentary diabetic rats than in control rats. The precursor form was significantly lower in diabetic exercised animals than in diabetic sedentary animals. However, the processed form did not change. These results demonstrate that exercise can modulate the muscle expression of MSTN protein in diabetic rats and suggest that MSTN may be involved in energy homeostasis.

Plasmids for variable expression of proteins targeted to the mitochondrial matrix or intermembrane space.

PubMed

Newman, Laura E; Schiavon, Cara; Kahn, Richard A

2016-01-01

We describe the construction and uses of a series of plasmids for directing expression to varied levels of exogenous proteins targeted to the mitochondrial matrix or intermembrane space. We found that the level of protein expression achieved, the kinetics of expression and mitochondrial import, and half-life after import can each vary with the protein examined. These factors should be considered when directing localization of an exogenous protein to mitochondria for rescue, proteomics, or other approaches. We describe the construction of a collection of plasmids for varied expression of proteins targeted to the mitochondrial matrix or intermembrane space, using previously defined targeting sequences and strength CMV promoters. The limited size of these compartments makes them particularly vulnerable to artifacts from over-expression. We found that different proteins display different kinetics of expression and import that should be considered when analyzing results from this approach. Finally, this collection of plasmids has been deposited in the Addgene plasmid repository to facilitate the ready access and use of these tools.

miR-24 and miR-205 expression is dependent on HPV onco-protein expression in keratinocytes

DOE Office of Scientific and Technical Information (OSTI.GOV)

McKenna, Declan J., E-mail: dj.mckenna@ulster.ac.uk; Centre for Cancer Research and Cell Biology, School of Medicine, Dentistry and Biomedical Science, Queen's University Belfast, Belfast BT9 7BL; Patel, Daksha, E-mail: d.patel@qub.ac.uk

2014-01-05

A screen of microRNA (miRNA) expression following differentiation in human foreskin keratinocytes (HFKs) identified changes in several miRNAs, including miR-24 and miR-205. We investigated how expression of Human Papilloma Virus Type-16 (HPV16) onco-proteins E6 and E7 affected expression of miR-24 and miR-205 during proliferation and differentiation of HFKs. We show that the induction of both miR-24 and miR-205 observed during differentiation of HFKs is lost in HFKs expressing E6 and E7. We demonstrate that the effect on miR-205 is due to E7 activity, as miR-205 expression is dependent on pRb expression. Finally, we provide evidence that miR-24 effects in themore » cell may be due to targeting of cyclin dependent kinase inhibitor p27. In summary, these results indicate that expression of both miR-24 and miR-205 are impacted by E6 and/or E7 expression, which may be one mechanism by which HPV onco-proteins can disrupt the balance between proliferation and differentiation in keratinocytes. - Highlights: • miR-24 and miR-205 are induced during keratinocyte differentiation. • This induction is lost in keratinocytes expressing HPV onco-proteins E6 and E7. • miR-205 is dependent upon pRb expression. • miR-24 targets p27 in cycling keratinocytes.« less

Fabrication and implantation of miniature dual-element strain gages for measuring in vivo gastrointestinal contractions in rodents.

PubMed

Holmes, Gregory M; Swartz, Emily M; McLean, Margaret S

2014-09-18

Gastrointestinal dysfunction remains a major cause of morbidity and mortality. Indeed, gastrointestinal (GI) motility in health and disease remains an area of productive research with over 1,400 published animal studies in just the last 5 years. Numerous techniques have been developed for quantifying smooth muscle activity of the stomach, small intestine, and colon. In vitro and ex vivo techniques offer powerful tools for mechanistic studies of GI function, but outside the context of the integrated systems inherent to an intact organism. Typically, measuring in vivo smooth muscle contractions of the stomach has involved an anesthetized preparation coupled with the introduction of a surgically placed pressure sensor, a static pressure load such as a mildly inflated balloon or by distending the stomach with fluid under barostatically-controlled feedback. Yet many of these approaches present unique disadvantages regarding both the interpretation of results as well as applicability for in vivo use in conscious experimental animal models. The use of dual element strain gages that have been affixed to the serosal surface of the GI tract has offered numerous experimental advantages, which may continue to outweigh the disadvantages. Since these gages are not commercially available, this video presentation provides a detailed, step-by-step guide to the fabrication of the current design of these gages. The strain gage described in this protocol is a design for recording gastric motility in rats. This design has been modified for recording smooth muscle activity along the entire GI tract and requires only subtle variation in the overall fabrication. Representative data from the entire GI tract are included as well as discussion of analysis methods, data interpretation and presentation.

The recombinant expression and activity detection of MAF-1 fusion protein.

PubMed

Fu, Ping; Wu, Jianwei; Gao, Song; Guo, Guo; Zhang, Yong; Liu, Jian

2015-10-01

This study establishes the recombinant expression system of MAF-1 (Musca domestica antifungal peptide-1) and demonstrates the antifungal activity of the expression product and shows the relationship between biological activity and structure. The gene segments on mature peptide part of MAF-1 were cloned, based on the primers designed according to the cDNA sequence of MAF-1. We constructed the recombinant prokaryotic expression plasmid using prokaryotic expression vector (pET-28a(+)) and converted it to the competent cell of BL21(DE3) to gain recombinant MAF-1 fusion protein with His tag sequence through purifying affinity chromatographic column of Ni-NTA. To conduct the Western Blotting test, recombinant MAF-1 fusion protein was used to produce the polyclonal antibody of rat. The antifungal activity of the expression product was detected using Candida albicans (ATCC10231) as the indicator. The MAF-1 recombinant fusion protein was purified to exhibit obvious antifungal activity, which lays the foundation for the further study of MAF-1 biological activity, the relationship between structure and function, as well as control of gene expression.

Quantitative Proteomic Analysis of Differentially Expressed Protein Profiles Involved in Pancreatic Ductal Adenocarcinoma

PubMed Central

Kuo, Kung-Kai; Kuo, Chao-Jen; Chiu, Chiang-Yen; Liang, Shih-Shin; Huang, Chun-Hao; Chi, Shu-Wen; Tsai, Kun-Bow; Chen, Chiao-Yun; Hsi, Edward; Cheng, Kuang-Hung; Chiou, Shyh-Horng

2016-01-01

Objectives The aim of this study was to identify differentially expressed proteins among various stages of pancreatic ductal adenocarcinoma (PDAC) by shotgun proteomics using nano-liquid chromatography coupled tandem mass spectrometry and stable isotope dimethyl labeling. Methods Differentially expressed proteins were identified and compared based on the mass spectral differences of their isotope-labeled peptide fragments generated from protease digestion. Results Our quantitative proteomic analysis of the differentially expressed proteins with stable isotope (deuterium/hydrogen ratio, ≥2) identified a total of 353 proteins, with at least 5 protein biomarker proteins that were significantly differentially expressed between cancer and normal mice by at least a 2-fold alteration. These 5 protein biomarker candidates include α-enolase, α-catenin, 14-3-3 β, VDAC1, and calmodulin with high confidence levels. The expression levels were also found to be in agreement with those examined by Western blot and histochemical staining. Conclusions The systematic decrease or increase of these identified marker proteins may potentially reflect the morphological aberrations and diseased stages of pancreas carcinoma throughout progressive developments leading to PDAC. The results would form a firm foundation for future work concerning validation and clinical translation of some identified biomarkers into targeted diagnosis and therapy for various stages of PDAC. PMID:26262590

Loads calibrations of strain gage bridges on the DAST project Aeroelastic Research Wing (ARW-2)

NASA Technical Reports Server (NTRS)

Eckstrom, C. V.

1986-01-01

Results from and details of the procedure used to calibrate strain gage bridges for measurements of wing structural loads, shear (V), bending moment (M), and torque (T), at three semispan stations on both the left and right semispans of the ARW-2 wing are presented. The ARW-2 wing has a reference area of 35 square feet, a span of 19 feet, an aspect ratio of 10.3, a midchord line sweepback angle of 25 degrees, and a taper ratio of 0.4. The ARW-2 wing was fabricated using aluminum spars and ribs covered with a fiberglass/honeycomb sandwich skin material. All strain gage bridges are mounted along with an estimate of their accuracy by means of a comparison of computed loads versus actual loads for three simulated flight conditions.

Protein-protein interaction network of gene expression in the hydrocortisone-treated keloid.

PubMed

Chen, Rui; Zhang, Zhiliang; Xue, Zhujia; Wang, Lin; Fu, Mingang; Lu, Yi; Bai, Ling; Zhang, Ping; Fan, Zhihong

2015-01-01

In order to explore the molecular mechanism of hydrocortisone in keloid tissue, the gene expression profiles of keloid samples treated with hydrocortisone were subjected to bioinformatics analysis. Firstly, the gene expression profiles (GSE7890) of five samples of keloid treated with hydrocortisone and five untreated keloid samples were downloaded from the Gene Expression Omnibus (GEO) database. Secondly, data were preprocessed using packages in R language and differentially expressed genes (DEGs) were screened using a significance analysis of microarrays (SAM) protocol. Thirdly, the DEGs were subjected to gene ontology (GO) function and KEGG pathway enrichment analysis. Finally, the interactions of DEGs in samples of keloid treated with hydrocortisone were explored in a human protein-protein interaction (PPI) network, and sub-modules of the DEGs interaction network were analyzed using Cytoscape software. Based on the analysis, 572 DEGs in the hydrocortisone-treated samples were screened; most of these were involved in the signal transduction and cell cycle. Furthermore, three critical genes in the module, including COL1A1, NID1, and PRELP, were screened in the PPI network analysis. These findings enhance understanding of the pathogenesis of the keloid and provide references for keloid therapy. © 2015 The International Society of Dermatology.

Mutational Analysis of the Rift Valley Fever Virus Glycoprotein Precursor Proteins for Gn Protein Expression

PubMed Central

Phoenix, Inaia; Lokugamage, Nandadeva; Nishiyama, Shoko; Ikegami, Tetsuro

2016-01-01

The Rift Valley fever virus (RVFV) M-segment encodes the 78 kD, NSm, Gn, and Gc proteins. The 1st AUG generates the 78 kD-Gc precursor, the 2nd AUG generates the NSm-Gn-Gc precursor, and the 3rd AUG makes the NSm’-Gn-Gc precursor. To understand biological changes due to abolishment of the precursors, we quantitatively measured Gn secretion using a reporter assay, in which a Gaussia luciferase (gLuc) protein is fused to the RVFV M-segment pre-Gn region. Using the reporter assay, the relative expression of Gn/gLuc fusion proteins was analyzed among various AUG mutants. The reporter assay showed efficient secretion of Gn/gLuc protein from the precursor made from the 2nd AUG, while the removal of the untranslated region upstream of the 2nd AUG (AUG2-M) increased the secretion of the Gn/gLuc protein. Subsequently, recombinant MP-12 strains encoding mutations in the pre-Gn region were rescued, and virological phenotypes were characterized. Recombinant MP-12 encoding the AUG2-M mutation replicated slightly less efficiently than the control, indicating that viral replication is further influenced by the biological processes occurring after Gn expression, rather than the Gn abundance. This study showed that, not only the abolishment of AUG, but also the truncation of viral UTR, affects the expression of Gn protein by the RVFV M-segment. PMID:27231931

Mutational Analysis of the Rift Valley Fever Virus Glycoprotein Precursor Proteins for Gn Protein Expression.

PubMed

Phoenix, Inaia; Lokugamage, Nandadeva; Nishiyama, Shoko; Ikegami, Tetsuro

2016-05-24

The Rift Valley fever virus (RVFV) M-segment encodes the 78 kD, NSm, Gn, and Gc proteins. The 1st AUG generates the 78 kD-Gc precursor, the 2nd AUG generates the NSm-Gn-Gc precursor, and the 3rd AUG makes the NSm'-Gn-Gc precursor. To understand biological changes due to abolishment of the precursors, we quantitatively measured Gn secretion using a reporter assay, in which a Gaussia luciferase (gLuc) protein is fused to the RVFV M-segment pre-Gn region. Using the reporter assay, the relative expression of Gn/gLuc fusion proteins was analyzed among various AUG mutants. The reporter assay showed efficient secretion of Gn/gLuc protein from the precursor made from the 2nd AUG, while the removal of the untranslated region upstream of the 2nd AUG (AUG2-M) increased the secretion of the Gn/gLuc protein. Subsequently, recombinant MP-12 strains encoding mutations in the pre-Gn region were rescued, and virological phenotypes were characterized. Recombinant MP-12 encoding the AUG2-M mutation replicated slightly less efficiently than the control, indicating that viral replication is further influenced by the biological processes occurring after Gn expression, rather than the Gn abundance. This study showed that, not only the abolishment of AUG, but also the truncation of viral UTR, affects the expression of Gn protein by the RVFV M-segment.

Differentially expressed proteins among normal cervix, cervical intraepithelial neoplasia and cervical squamous cell carcinoma.

PubMed

Zhao, Q; He, Y; Wang, X-L; Zhang, Y-X; Wu, Y-M

2015-08-01

To explore the differentially expressed proteins in normal cervix, cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma (CSCC) tissues by differential proteomics technique. Cervical tissues (including normal cervix, CIN and CSCC) were collected in Department of Gynecologic Oncology of Beijing Obstetrics and Gynecology Hospital. Two-dimensional fluorescence difference in gel electrophoresis (2-D DIGE) and DeCyder software were used to detect the differentially expressed proteins. Matrix-assisted laser desorption/ionization-time-of-flight tandem mass spectrometry (MALDI-TOF/TOF MS) was used to identify the differentially expressed proteins. Western blot (WB) and immunohistochemistry (IHC) were performed to validate the expressions of selected proteins among normal cervix, CIN and CSCC. 2-D DIGE images with high resolution and good repeatability were obtained. Forty-six differentially expressed proteins (27 up-regulated and 19 down-regulated) were differentially expressed among the normal cervix, CIN and CSCC. 26 proteins were successfully identified by MALDI-TOF/TOF MS. S100A9 (S100 calcium-binding protein A9) was the most significantly up-regulated protein. Eukaryotic elongation factor 1-alpha-1 (eEF1A1) was the most significantly down-regulated protein. Pyruvate kinase isozymes M2 (PKM2) was both up-regulated and down-regulated. The results of WB showed that with the increase in the severity of cervical lesions, the expression of S100A9 protein was significantly increased among the three groups (P = 0.010). The expression of eEF1A1 was reduced but without significant difference (P = 0.861). The expression of PKM2 was significantly reduced (P = 0.000). IHC showed that protein S100A9 was mainly expressed in the cytoplasm, and its positive expression rate was 20.0 % in normal cervix, 70.0 % in CIN and 100.0 % in CSCC, with a significant difference among them (P = 0.006). eEF1A1 was mainly expressed in the cell plasma, and its

Human trabecular meshwork cells express BMP antagonist mRNAs and proteins.

PubMed

Tovar-Vidales, Tara; Fitzgerald, Ashley M; Clark, Abbot F

2016-06-01

Glaucoma patients have elevated aqueous humor and trabecular meshwork (TM) levels of transforming growth factor-beta2 (TGF-β2). TGF-β2 has been associated with increased extracellular matrix (ECM) deposition (i.e. fibronectin), which is attributed to the increased resistance of aqueous humor outflow through the TM. We have previously demonstrated that bone morphogenetic protein (BMP) 4 selectively counteracts the profibrotic effect of TGF-β2 with respect to ECM synthesis in the TM, and this action is reversed by the BMP antagonist gremlin. Thus, the BMP and TGF-β signaling pathways antagonize each other's antifibrotic and profibrotic roles. The purpose of this study was to determine whether cultured human TM cells: (a) express other BMP antagonists including noggin, chordin, BMPER, BAMBI, Smurf1 and 2, and (b) whether expression of these proteins is regulated by exogenous TGF-β2 treatment. Primary human trabecular meshwork (TM) cells were grown to confluency and treated with TGF-β2 (5 ng/ml) for 24 or 48 h in serum-free medium. Untreated cell served as controls. qPCR and Western immunoblots (WB) determined that human TM cells expressed mRNAs and proteins for the BMP antagonist proteins: noggin, chordin, BMPER, BAMBI, and Smurf1/2. Exogenous TGF-β2 decreased chordin, BMPER, BAMBI, and Smurf1 mRNA and protein expression. In contrast, TGF-β2 increased secreted noggin and Smurf2 mRNA and protein levels. BMP antagonist members are expressed in the human TM. These molecules may be involved in the normal function of the TM as well as TM pathogenesis. Altered expression of BMP antagonist members may lead to functional changes in the human TM. Copyright © 2016 Elsevier Ltd. All rights reserved.

SGLT2 Protein Expression Is Increased in Human Diabetic Nephropathy

PubMed Central

Wang, Xiaoxin X.; Levi, Jonathan; Luo, Yuhuan; Myakala, Komuraiah; Herman-Edelstein, Michal; Qiu, Liru; Wang, Dong; Peng, Yingqiong; Grenz, Almut; Lucia, Scott; Dobrinskikh, Evgenia; D'Agati, Vivette D.; Koepsell, Hermann; Kopp, Jeffrey B.; Rosenberg, Avi Z.; Levi, Moshe

2017-01-01

There is very limited human renal sodium gradient-dependent glucose transporter protein (SGLT2) mRNA and protein expression data reported in the literature. The first aim of this study was to determine SGLT2 mRNA and protein levels in human and animal models of diabetic nephropathy. We have found that the expression of SGLT2 mRNA and protein is increased in renal biopsies from human subjects with diabetic nephropathy. This is in contrast to db-db mice that had no changes in renal SGLT2 protein expression. Furthermore, the effect of SGLT2 inhibition on renal lipid content and inflammation is not known. The second aim of this study was to determine the potential mechanisms of beneficial effects of SGLT2 inhibition in the progression of diabetic renal disease. We treated db/db mice with a selective SGLT2 inhibitor JNJ 39933673. We found that SGLT2 inhibition caused marked decreases in systolic blood pressure, kidney weight/body weight ratio, urinary albumin, and urinary thiobarbituric acid-reacting substances. SGLT2 inhibition prevented renal lipid accumulation via inhibition of carbohydrate-responsive element-binding protein-β, pyruvate kinase L, SCD-1, and DGAT1, key transcriptional factors and enzymes that mediate fatty acid and triglyceride synthesis. SGLT2 inhibition also prevented inflammation via inhibition of CD68 macrophage accumulation and expression of p65, TLR4, MCP-1, and osteopontin. These effects were associated with reduced mesangial expansion, accumulation of the extracellular matrix proteins fibronectin and type IV collagen, and loss of podocyte markers WT1 and synaptopodin, as determined by immunofluorescence microscopy. In summary, our study showed that SGLT2 inhibition modulates renal lipid metabolism and inflammation and prevents the development of nephropathy in db/db mice. PMID:28196866

Rapid synthesis of DNA-cysteine conjugates for expressed protein ligation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lovrinovic, Marina; Niemeyer, Christof M.

2005-09-30

We report a rapid method for the covalent modification of commercially available amino-modified DNA oligonucleotides with a cysteine moiety. The resulting DNA-cysteine conjugates are versatile reagents for the efficient preparation of covalent DNA-protein conjugates by means of expressed protein ligation (EPL). The EPL method allows for the site-specific coupling of cysteine-modified DNA oligomers with recombinant intein-fusion proteins, the latter of which contain a C-terminal thioester enabling the mild and highly specific reaction with N-terminal cysteine compounds. We prepared a cysteine-modifier reagent in a single-step reaction which allows for the rapid and near quantitative synthesis of cysteine-DNA conjugates. The latter weremore » ligated with the green fluorescent protein mutant EYFP, recombinantly expressed as an intein-fusion protein, allowing for the mild and selective formation of EYFP-DNA conjugates in high yields of about 60%. We anticipate many applications of our approach, ranging from protein microarrays to the arising field of nanobiotechnology.« less

Cloning, Expression, and Purification of Brucella suis Outer Membrane Proteins

DTIC Science & Technology

2005-01-01

13-09-20061 4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER Cloning, expression and purification of Brucella suis outer membrane proteins 5b. GRANT NUMBER...attractive for this purpose. In this study, we cloned, expressed and purified seven predicted OMPs of Brucella suis . The recombinant proteins were...fused with 6-his and V5 epitope tags at their C termini to facilitate detection and purification. The B. suis surface genes were PCR synthesized based

The E4 protein; structure, function and patterns of expression

DOE Office of Scientific and Technical Information (OSTI.GOV)

Doorbar, John, E-mail: jdoorba@nimr.mrc.ac.uk

2013-10-15

The papillomavirus E4 open reading frame (ORF) is contained within the E2 ORF, with the primary E4 gene-product (E1{sup ∧}E4) being translated from a spliced mRNA that includes the E1 initiation codon and adjacent sequences. E4 is located centrally within the E2 gene, in a region that encodes the E2 protein′s flexible hinge domain. Although a number of minor E4 transcripts have been reported, it is the product of the abundant E1{sup ∧}E4 mRNA that has been most extensively analysed. During the papillomavirus life cycle, the E1{sup ∧}E4 gene products generally become detectable at the onset of vegetative viral genomemore » amplification as the late stages of infection begin. E4 contributes to genome amplification success and virus synthesis, with its high level of expression suggesting additional roles in virus release and/or transmission. In general, E4 is easily visualised in biopsy material by immunostaining, and can be detected in lesions caused by diverse papillomavirus types, including those of dogs, rabbits and cattle as well as humans. The E4 protein can serve as a biomarker of active virus infection, and in the case of high-risk human types also disease severity. In some cutaneous lesions, E4 can be expressed at higher levels than the virion coat proteins, and can account for as much as 30% of total lesional protein content. The E4 proteins of the Beta, Gamma and Mu HPV types assemble into distinctive cytoplasmic, and sometimes nuclear, inclusion granules. In general, the E4 proteins are expressed before L2 and L1, with their structure and function being modified, first by kinases as the infected cell progresses through the S and G2 cell cycle phases, but also by proteases as the cell exits the cell cycle and undergoes true terminal differentiation. The kinases that regulate E4 also affect other viral proteins simultaneously, and include protein kinase A, Cyclin-dependent kinase, members of the MAP Kinase family and protein kinase C. For HPV16 E1

Broad host range vectors for expression of proteins with (Twin-) Strep-tag, His-tag and engineered, export optimized yellow fluorescent protein

PubMed Central

2013-01-01

Background In current protein research, a limitation still is the production of active recombinant proteins or native protein associations to assess their function. Especially the localization and analysis of protein-complexes or the identification of modifications and small molecule interaction partners by co-purification experiments requires a controllable expression of affinity- and/or fluorescence tagged variants of a protein of interest in its native cellular background. Advantages of periplasmic and/or homologous expressions can frequently not be realized due to a lack of suitable tools. Instead, experiments are often limited to the heterologous production in one of the few well established expression strains. Results Here, we introduce a series of new RK2 based broad host range expression plasmids for inducible production of affinity- and fluorescence tagged proteins in the cytoplasm and periplasm of a wide range of Gram negative hosts which are designed to match the recently suggested modular Standard European Vector Architecture and database. The vectors are equipped with a yellow fluorescent protein variant which is engineered to fold and brightly fluoresce in the bacterial periplasm following Sec-mediated export, as shown from fractionation and imaging studies. Expression of Strep-tag®II and Twin-Strep-tag® fusion proteins in Pseudomonas putida KT2440 is demonstrated for various ORFs. Conclusion The broad host range constructs we have produced enable good and controlled expression of affinity tagged protein variants for single-step purification and qualify for complex co-purification experiments. Periplasmic export variants enable production of affinity tagged proteins and generation of fusion proteins with a novel engineered Aequorea-based yellow fluorescent reporter protein variant with activity in the periplasm of the tested Gram-negative model bacteria Pseudomonas putida KT2440 and Escherichia coli K12 for production, localization or co

Evaluation of a Crack-Growth Gage for Monitoring Possible Structural Fatigue-Crack Growth

DTIC Science & Technology

1978-02-01

the gages and structural components and tested the specimens. Appreciation is also extended to Dennis E. Macha for his effort in making the laser...SDM Conference AIAA/ASME, San Diego, California, March 21-23, 1977. 20 AFML-TR-77-233 REFERENCES (CONTINUED) 12. D. E. Macha , W. N. Sharpe, Jr., and A

Heterogeneity mapping of protein expression in tumors using quantitative immunofluorescence.

PubMed

Faratian, Dana; Christiansen, Jason; Gustavson, Mark; Jones, Christine; Scott, Christopher; Um, InHwa; Harrison, David J

2011-10-25

Morphologic heterogeneity within an individual tumor is well-recognized by histopathologists in surgical practice. While this often takes the form of areas of distinct differentiation into recognized histological subtypes, or different pathological grade, often there are more subtle differences in phenotype which defy accurate classification (Figure 1). Ultimately, since morphology is dictated by the underlying molecular phenotype, areas with visible differences are likely to be accompanied by differences in the expression of proteins which orchestrate cellular function and behavior, and therefore, appearance. The significance of visible and invisible (molecular) heterogeneity for prognosis is unknown, but recent evidence suggests that, at least at the genetic level, heterogeneity exists in the primary tumor(1,2), and some of these sub-clones give rise to metastatic (and therefore lethal) disease. Moreover, some proteins are measured as biomarkers because they are the targets of therapy (for instance ER and HER2 for tamoxifen and trastuzumab (Herceptin), respectively). If these proteins show variable expression within a tumor then therapeutic responses may also be variable. The widely used histopathologic scoring schemes for immunohistochemistry either ignore, or numerically homogenize the quantification of protein expression. Similarly, in destructive techniques, where the tumor samples are homogenized (such as gene expression profiling), quantitative information can be elucidated, but spatial information is lost. Genetic heterogeneity mapping approaches in pancreatic cancer have relied either on generation of a single cell suspension(3), or on macrodissection(4). A recent study has used quantum dots in order to map morphologic and molecular heterogeneity in prostate cancer tissue(5), providing proof of principle that morphology and molecular mapping is feasible, but falling short of quantifying the heterogeneity. Since immunohistochemistry is, at best, only semi

Expression and Localization of CLC Chloride Transport Proteins in the Avian Retina

PubMed Central

McMains, Emily; Krishnan, Vijai; Prasad, Sujitha; Gleason, Evanna

2011-01-01

Members of the ubiquitously expressed CLC protein family of chloride channels and transporters play important roles in regulating cellular chloride and pH. The CLCs that function as Cl−/H+ antiporters, ClCs 3–7, are essential in particular for the acidification of endosomal compartments and protein degradation. These proteins are broadly expressed in the nervous system, and mutations that disrupt their expression are responsible for several human genetic diseases. Furthermore, knock-out of ClC3 and ClC7 in the mouse result in the degeneration of the hippocampus and the retina. Despite this evidence of their importance in retinal function, the expression patterns of different CLC transporters in different retinal cell types are as yet undescribed. Previous work in our lab has shown that in chicken amacrine cells, internal Cl− can be dynamic. To determine whether CLCs have the potential to participate, we used PCR and immunohistochemical techniques to examine CLC transporter expression in the chicken retina. We observed a high level of variation in the retinal expression levels and patterns among the different CLC proteins examined. These findings, which represent the first systematic investigation of CLC transporter expression in the retina, support diverse functions for the different CLCs in this tissue. PMID:21408174

Functional evaluation of candidate ice structuring proteins using cell-free expression systems.

PubMed

Brödel, A K; Raymond, J A; Duman, J G; Bier, F F; Kubick, S

2013-02-10

Ice structuring proteins (ISPs) protect organisms from damage or death by freezing. They depress the non-equilibrium freezing point of water and prevent recrystallization, probably by binding to the surface of ice crystals. Many ISPs have been described and it is likely that many more exist in nature that have not yet been identified. ISPs come in many forms and thus cannot be reliably identified by their structure or consensus ice-binding motifs. Recombinant protein expression is the gold standard for proving the activity of a candidate ISP. Among existing expression systems, cell-free protein expression is the simplest and gives the fastest access to the protein of interest, but selection of the appropriate cell-free expression system is crucial for functionality. Here we describe cell-free expression methods for three ISPs that differ widely in structure and glycosylation status from three organisms: a fish (Macrozoarces americanus), an insect (Dendroides canadensis) and an alga (Chlamydomonas sp. CCMP681). We use both prokaryotic and eukaryotic expression systems for the production of ISPs. An ice recrystallization inhibition assay is used to test functionality. The techniques described here should improve the success of cell-free expression of ISPs in future applications. Copyright © 2012 Elsevier B.V. All rights reserved.

Expression of small heat shock proteins from pea seedlings under gravity altered conditions

NASA Astrophysics Data System (ADS)

Talalaev, Alexandr S.

2005-08-01

A goal of our study was to evaluate the stress gene expression in Pisum sativum seedlings exposed to altered gravity and temperature elevation. We investigate message for the two inducible forms of the cytosolic small heat shock proteins (sHsp), sHsp 17.7 and sHsp 18.1. Both proteins are able to enhance the refolding of chemically denatured proteins in an ATP- independent manner, in other words they can function as molecular chaperones. We studied sHsps expression in pea seedlings cells by Western blotting. Temperature elevation, as the positive control, significantly increased PsHsp 17.7 and PsHsp 18.1 expression. Expression of the housekeeping protein, actin was constant and comparable to unstressed controls for all treatments. We concluded that gravitational perturbations incurred by clinorotation did not change sHsp genes expression.

Discharge measurements at gaging stations

USGS Publications Warehouse

Turnipseed, D. Phil; Sauer, Vernon B.

2010-01-01

The techniques and standards for making discharge measurements at streamflow gaging stations are described in this publication. The vertical axis rotating-element current meter, principally the Price current meter, has been traditionally used for most measurements of discharge; however, advancements in acoustic technology have led to important developments in the use of acoustic Doppler current profilers, acoustic Doppler velocimeters, and other emerging technologies for the measurement of discharge. These new instruments, based on acoustic Doppler theory, have the advantage of no moving parts, and in the case of the acoustic Doppler current profiler, quickly and easily provide three-dimensional stream-velocity profile data through much of the vertical water column. For much of the discussion of acoustic Doppler current profiler moving-boat methodology, the reader is referred to U.S. Geological Survey Techniques and Methods 3-A22 (Mueller and Wagner, 2009). Personal digital assistants (PDAs), electronic field notebooks, and other personal computers provide fast and efficient data-collection methods that are more error-free than traditional hand methods. The use of portable weirs and flumes, floats, volumetric tanks, indirect methods, and tracers in measuring discharge are briefly described.

The Atmospheric Lifetime Experiment and the Global Atmospheric Gas Experiment (ALE/GAGE)