The Design and Analysis of Salmonid Tagging Studies in the Columbia Basin : Volume II: Experiment Salmonid Survival with Combined PIT-CWT Tagging.

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

Newman, Ken

1997-06-01

Experiment designs to estimate the effect of transportation on survival and return rates of Columbia River system salmonids are discussed along with statistical modeling techniques. Besides transportation, river flow and dam spill are necessary components in the design and analysis otherwise questions as to the effects of reservoir drawdowns and increased dam spill may never be satisfactorily answered. Four criteria for comparing different experiment designs are: (1) feasibility, (2) clarity of results, (3) scope of inference, and (4) time to learn. In this report, alternative designs for conducting experimental manipulations of smolt tagging studies to study effects of river operationsmore » such as flow levels, spill fractions, and transporting outmigrating salmonids around dams in the Columbia River system are presented. The principles of study design discussed in this report have broad implications for the many studies proposed to investigate both smolt and adult survival relationships. The concepts are illustrated for the case of the design and analysis of smolt transportation experiments. The merits of proposed transportation studies should be measured relative to these principles of proper statistical design and analysis.« less

Comparison of a simulated velocity profile of a turbulent boundary layer with measurements obtained by Femtosecond Laser Electronic Excitation Tagging (FLEET)

NASA Astrophysics Data System (ADS)

New-Tolley, Matthew; Zhang, Yibin; Shneider, Mikhail; Miles, Richard

2017-11-01

Accurate velocimetry measurements of turbulent flows are essential for improving our understanding of turbulent phenomena and validating numerical approaches. Femtosecond Laser Electronic Excitation Tagging (FLEET) is an unseeded molecular tagging method for velocimetry measurements in flows which contain nitrogen. A femtosecond laser pulse is used to ionize and dissociate nitrogen molecules within its focal zone. The decaying plasma fluoresces in the visible and infrared spectrum over a period of microseconds which allows the displacement of the tagged region to be photographed to determine velocity. This study compares the experimental and numerical advection of the tagged region in a turbulent boundary layer generated by a supersonic flow over a flat plate. The tagged region in the simulation is approximated as an infinitely thin cylinder while the flow field is generated using the steady state boundary layer equations with an algebraic turbulence model. This approximation is justified by previous computational analyses, using an unsteady three-dimensional Navier-Stokes solver, which indicate that the radial perturbations of the tagged region are negligible compared to its translation. This research was conducted with government support from the Air Force Office of Scientific Research under Dr. Ivett Leyva and the Army Research Office under Dr. Matthew Munson.

Tag-to-Tag Interference Suppression Technique Based on Time Division for RFID.

PubMed

Khadka, Grishma; Hwang, Suk-Seung

2017-01-01

Radio-frequency identification (RFID) is a tracking technology that enables immediate automatic object identification and rapid data sharing for a wide variety of modern applications using radio waves for data transmission from a tag to a reader. RFID is already well established in technical areas, and many companies have developed corresponding standards and measurement techniques. In the construction industry, effective monitoring of materials and equipment is an important task, and RFID helps to improve monitoring and controlling capabilities, in addition to enabling automation for construction projects. However, on construction sites, there are many tagged objects and multiple RFID tags that may interfere with each other's communications. This reduces the reliability and efficiency of the RFID system. In this paper, we propose an anti-collision algorithm for communication between multiple tags and a reader. In order to suppress interference signals from multiple neighboring tags, the proposed algorithm employs the time-division (TD) technique, where tags in the interrogation zone are assigned a specific time slot so that at every instance in time, a reader communicates with tags using the specific time slot. We present representative computer simulation examples to illustrate the performance of the proposed anti-collision technique for multiple RFID tags.

Mathematical Development and Computational Analysis of Harmonic Phase-Magnetic Resonance Imaging (HARP-MRI) Based on Bloch Nuclear Magnetic Resonance (NMR) Diffusion Model for Myocardial Motion.

PubMed

Dada, Michael O; Jayeoba, Babatunde; Awojoyogbe, Bamidele O; Uno, Uno E; Awe, Oluseyi E

2017-09-13

Harmonic Phase-Magnetic Resonance Imaging (HARP-MRI) is a tagged image analysis method that can measure myocardial motion and strain in near real-time and is considered a potential candidate to make magnetic resonance tagging clinically viable. However, analytical expressions of radially tagged transverse magnetization in polar coordinates (which is required to appropriately describe the shape of the heart) have not been explored because the physics required to directly connect myocardial deformation of tagged Nuclear Magnetic Resonance (NMR) transverse magnetization in polar geometry and the appropriate harmonic phase parameters are not yet available. The analytical solution of Bloch NMR diffusion equation in spherical geometry with appropriate spherical wave tagging function is important for proper analysis and monitoring of heart systolic and diastolic deformation with relevant boundary conditions. In this study, we applied Harmonic Phase MRI method to compute the difference between tagged and untagged NMR transverse magnetization based on the Bloch NMR diffusion equation and obtained radial wave tagging function for analysis of myocardial motion. The analytical solution of the Bloch NMR equations and the computational simulation of myocardial motion as developed in this study are intended to significantly improve healthcare for accurate diagnosis, prognosis and treatment of cardiovascular related deceases at the lowest cost because MRI scan is still one of the most expensive anywhere. The analysis is fundamental and significant because all Magnetic Resonance Imaging techniques are based on the Bloch NMR flow equations.

Application of the hydroxyl tagging velocimetry to direct-connect supersonic combustor experiment

NASA Astrophysics Data System (ADS)

Ye, Jingfeng; Li, Guohua; Shao, Jun; Hu, Zhiyun; Zhao, Xinyan; Song, WenYan

2017-05-01

For the purpose of measuring the flow velocity in a scramjet test model, an special designed measurement system was established, including the strong vibration suppression, optical transport consideration, the movable device etc. The interference of the strong vibration to the velocity measurements was avoided by two ICCD cameras capturing the reference tag lines image and moved tag lines image together during an experiment. According to the tag lines image feature, data processing including correlation algorithm, data fitting by a Gauss function were used respectively to extract the positions of the reference tag lines and the moved tag lines. The velocity measurements were carried out at the isolation section and the cavity section. The results showed that the well SNR could be achieved in the H2/air combustion heating flow, but in the kerosene fuel combustion flow, the measurements images might be interfered by the strong OH background from the chemical reaction, and the signal intensity could be reduced due to the tag laser attenuation through the absorption by kerosene vapor. But when the combustor model was run at a low chemical equivalent, the interference could be suppressed to an accepted level.

Evaluation of stream flow effects on smolt survival in the Yakima River Basin, Washington, 2012-2014

USGS Publications Warehouse

Courter, Ian; Garrison, Tommy; Kock, Tobias J.; Perry, Russell W.

2015-01-01

The influence of stream flow on survival of emigrating juvenile (smolts) Pacific salmon Oncorhynchus spp. and steelhead trout O. mykiss is of key management interest. However, few studies have quantified flow effects on smolt migration survival, and available information does not indicate a consistent flow-survival relationship within the typical range of flows under management control. It is hypothesized that smolt migration and dam passage survival are positively correlated with stream flow because higher flows increase migration rates, potentially reducing exposure to predation, and reduce delays in reservoirs. However, available empirical data are somewhat equivocal concerning the influence of flow on smolt survival and the underlying mechanisms driving this relationship. Stream flow effects on survival of emigrating anadromous salmonids in the Yakima Basin have concerned water users and fisheries managers for over 20 years, and previous studies do not provide sufficient information at the resolution necessary to inform water operations, which typically occur on a small spatiotemporal scale. Using a series of controlled flow releases from 2012-2014, combined with radio telemetry, we quantified the relationship between flow and smolt survival from Roza Dam 208 km downstream to the Yakima River mouth, as well as for specific routes of passage at Roza Dam. A novel multistate mark-recapture model accounted for weekly variation in flow conditions experienced by radio-tagged fish. Groups of fish were captured and radio-tagged at Roza Dam and released at two locations, upstream at the Big Pines Campground (river kilometer [rkm] 211) and downstream in the Roza Dam tailrace (rkm 208). A total of 904 hatchery-origin yearling Chinook salmon O. tshawytscha were captured in the Roza Dam fish bypass, radio-tagged and released upstream of Roza Dam. Two hundred thirty seven fish were released in the tailrace of Roza Dam. Fish released in the tailrace of Roza Dam were tagged concurrently with fish released upstream of the dam using identical tagging methods. Tagging and release events were conducted to target a range of flow conditions indicative of flows observed during the typical migration period (March-May) for juvenile spring Chinook salmon in the Yakima River. Three, five and four separate upstream releases were conducted in 2012, 2013, and 2014 respectively, and at least 43 fish were released alive on each occasion. The release sample sizes in 2014 were much larger (~130) compared to previous years for the purpose of increasing precision of survival estimates across the range of flows tested. Migration movements of radio-tagged spring Chinook salmon smolts were monitored with an array of telemetry receiver stations (fixed sites) that extended 208 rkm downstream from the forebay of Roza Dam to the mouth of the Yakima River. Fixed monitoring sites included the forebay of Roza Dam (rkm 208), the tailrace of Roza Dam (rkm 207.9), the mouth of Wenas Creek (rkm 199.2), the mouth of the Naches River (two sites, rkm 189.4), Sunnyside Dam (two sites, rkm 169.1), Prosser Dam (rkm 77.2), and the mouth of the Yakima River (two sites, rkm2 3). This array segregated the study area into four discrete reaches in which survival of tagged fish was estimated. Aerial and underwater antennas were also used to monitor tagged fish at Roza Dam. Aerial antennas were located in the forebay, on the East gate, on the West gate, and in the tailrace of Roza Dam. Underwater antennas were located in the fish bypass, upstream of the East gate, and upstream of the West gate to collect route-specific passage data for tagged fish. Additional years of data collection and analysis could alter or improve our understanding of the influence of flow and other environmental factors on smolt survival in the Yakima River. Nevertheless, during 2012-2014, yearling hatchery Chinook salmon smolt emigration survival was significantly associated with stream flow in the

Development of RAP Tag, a Novel Tagging System for Protein Detection and Purification.

PubMed

Fujii, Yuki; Kaneko, Mika K; Ogasawara, Satoshi; Yamada, Shinji; Yanaka, Miyuki; Nakamura, Takuro; Saidoh, Noriko; Yoshida, Kanae; Honma, Ryusuke; Kato, Yukinari

2017-04-01

Affinity tag systems, possessing high affinity and specificity, are useful for protein detection and purification. The most suitable tag for a particular purpose should be selected from many available affinity tag systems. In this study, we developed a novel affinity tag called the "RAP tag" system, which comprises a mouse antirat podoplanin monoclonal antibody (clone PMab-2) and the RAP tag (DMVNPGLEDRIE). This system is useful not only for protein detection in Western blotting, flow cytometry, and sandwich enzyme-linked immunosorbent assay, but also for protein purification.

Enhanced acetyl-CoA production is associated with increased triglyceride accumulation in the green alga Chlorella desiccata

PubMed Central

Avidan, Omri; Brandis, Alexander; Rogachev, Ilana; Pick, Uri

2015-01-01

Triglycerides (TAGs) from microalgae can be utilized as food supplements and for biodiesel production, but little is known about the regulation of their biosynthesis. This work aimed to test the relationship between acetyl-CoA (Ac-CoA) levels and TAG biosynthesis in green algae under nitrogen deprivation. A novel, highly sensitive liquid chromatography mass spectrometry (LC-MS/MS) technique enabled us to determine the levels of Ac-CoA, malonyl-CoA, and unacetylated (free) CoA in green microalgae. A comparative study of three algal species that differ in TAG accumulation levels shows that during N starvation, Ac-CoA levels rapidly rise, preceding TAG accumulation in all tested species. The levels of Ac-CoA in the high TAG accumulator Chlorella desiccata exceed the levels in the moderate TAG accumulators Dunaliella tertiolecta and Chlamydomonas reinhardtii. Similarly, malonyl-CoA and free CoA levels also increase, but to lower extents. Calculated cellular concentrations of Ac-CoA are far lower than reported K mAc-CoA values of plastidic Ac-CoA carboxylase (ptACCase) in plants. Transcript level analysis of plastidic pyruvate dehydrogenase (ptPDH), the major chloroplastic Ac-CoA producer, revealed rapid induction in parallel with Ac-CoA accumulation in C. desiccata, but not in D. tertiolecta or C. reinhardtii. It is proposed that the capacity to accumulate high TAG levels in green algae critically depends on their ability to divert carbon flow towards Ac-CoA. This requires elevation of the chloroplastic CoA pool level and enhancement of Ac-CoA biosynthesis. These conclusions may have important implications for future genetic manipulation to enhance TAG biosynthesis in green algae. PMID:25922486

Technique for the application of a streamer-type fish tag

USGS Publications Warehouse

Joeris, Leonard S.

1953-01-01

Principal features of the technique are: attachment of the plastic tag by means of a nylon-thread loop prepared in advance of field work; use of a curved surgical needle with cutting edge and a split eye for application of the tag. The procedures for splitting the needle's eye and for applying the tag are described and illustrated by a series of photographs.

Numerical and experimental hydrodynamic analysis of suction cup bio-logging tag designs for marine mammals

NASA Astrophysics Data System (ADS)

Murray, Mark; Shorter, Alex; Howle, Laurens; Johnson, Mark; Moore, Michael

2012-11-01

The improvement and miniaturization of sensing technologies has made bio-logging tags, utilized for the study of marine mammal behavior, more practical. These sophisticated sensing packages require a housing which protects the electronics from the environment and provides a means of attachment to the animal. The hydrodynamic forces on these housings can inadvertently remove the tag or adversely affect the behavior or energetics of the animal. A modification to the original design of a suction cup bio-logging tag housing was desired to minimize the adverse forces. In this work, hydrodynamic loading of two suction cup tag designs, original and modified designs, were analyzed using computational fluid dynamics (CFD) models and validated experimentally. Overall, the simulation and experimental results demonstrated that a tag housing that minimized geometric disruptions to the flow reduced drag forces, and that a tag housing with a small frontal cross-sectional area close to the attachment surface reduced lift forces. Preliminary results from experimental work with a common dolphin cadaver indicates that the suction cups used to attach the tags to the animal provide sufficient attachment force to resist failure at predicted drag and lift forces in 10 m/s flow.

Molecular candidates of MTV in air

NASA Astrophysics Data System (ADS)

Dam, Nico; Mirzaei, Mehrnoosh; van de Water, Willem

2011-11-01

In molecular tagging velocimetry (MTV), the molecules of a gas are used as flow tracers. These tracers can be produced at will by illumination with a laser which promotes molecules to a long- lived excited state, fuses N2 and N2 to NO, or makes molecules phosphoresce. A while later these tagged molecules can be visualized by laser-induced fluorescence, or by just watching them while they phosphoresce. Candidates for MTV in turbulence research must be arranged in structures narrower than the Kolmogorov scale, which remain narrow as time progresses, and must live longer than the Kolmogorov time. These requirements invalidate many candidates, candidates once deemed successful. They do so in various surprising manners that involve a combination of fluid flow and molecular dynamics. Rather than velocimetry in turbulence, MTV techniques offer a unique view on basic dispersion processes at the smallest scales of turbulence. In this way we have measured the spreading of clouds whose size is a few times the Kolmogorov length and the Batchelor dispersion of objects whose size is inside the inertial range.

Flow cytometry-assisted rapid isolation of recombinant Plasmodium berghei parasites exemplified by functional analysis of aquaglyceroporin

PubMed Central

Kenthirapalan, Sanketha; Waters, Andrew P.; Matuschewski, Kai; Kooij, Taco W.A.

2012-01-01

The most critical bottleneck in the generation of recombinant Plasmodium berghei parasites is the mandatory in vivo cloning step following successful genetic manipulation. This study describes a new technique for rapid selection of recombinant P. berghei parasites. The method is based on flow cytometry to isolate isogenic parasite lines and represents a major advance for the field, in that it will speed the generation of recombinant parasites as well as cut down on animal use significantly. High expression of GFP during blood infection, a prerequisite for robust separation of transgenic lines by flow cytometry, was achieved. Isogenic recombinant parasite populations were isolated even in the presence of a 100-fold excess of wild-type (WT) parasites. Aquaglyceroporin (AQP) loss-of-function mutants and parasites expressing a tagged AQP were generated to validate this approach. aqp− parasites grow normally within the WT phenotypic range during blood infection of NMRI mice. Similarly, colonization of the insect vector and establishment of an infection after mosquito transmission were unaffected, indicating that AQP is dispensable for life cycle progression in vivo under physiological conditions, refuting its use as a suitable drug target. Tagged AQP localized to perinuclear structures and not the parasite plasma membrane. We suggest that flow-cytometric isolation of isogenic parasites overcomes the major roadblock towards a genome-scale repository of mutant and transgenic malaria parasite lines. PMID:23137753

Myocardial tagging by Cardiovascular Magnetic Resonance: evolution of techniques--pulse sequences, analysis algorithms, and applications

PubMed Central

2011-01-01

Cardiovascular magnetic resonance (CMR) tagging has been established as an essential technique for measuring regional myocardial function. It allows quantification of local intramyocardial motion measures, e.g. strain and strain rate. The invention of CMR tagging came in the late eighties, where the technique allowed for the first time for visualizing transmural myocardial movement without having to implant physical markers. This new idea opened the door for a series of developments and improvements that continue up to the present time. Different tagging techniques are currently available that are more extensive, improved, and sophisticated than they were twenty years ago. Each of these techniques has different versions for improved resolution, signal-to-noise ratio (SNR), scan time, anatomical coverage, three-dimensional capability, and image quality. The tagging techniques covered in this article can be broadly divided into two main categories: 1) Basic techniques, which include magnetization saturation, spatial modulation of magnetization (SPAMM), delay alternating with nutations for tailored excitation (DANTE), and complementary SPAMM (CSPAMM); and 2) Advanced techniques, which include harmonic phase (HARP), displacement encoding with stimulated echoes (DENSE), and strain encoding (SENC). Although most of these techniques were developed by separate groups and evolved from different backgrounds, they are in fact closely related to each other, and they can be interpreted from more than one perspective. Some of these techniques even followed parallel paths of developments, as illustrated in the article. As each technique has its own advantages, some efforts have been made to combine different techniques together for improved image quality or composite information acquisition. In this review, different developments in pulse sequences and related image processing techniques are described along with the necessities that led to their invention, which makes this article easy to read and the covered techniques easy to follow. Major studies that applied CMR tagging for studying myocardial mechanics are also summarized. Finally, the current article includes a plethora of ideas and techniques with over 300 references that motivate the reader to think about the future of CMR tagging. PMID:21798021

High Throughput Biological Analysis Using Multi-bit Magnetic Digital Planar Tags

NASA Astrophysics Data System (ADS)

Hong, B.; Jeong, J.-R.; Llandro, J.; Hayward, T. J.; Ionescu, A.; Trypiniotis, T.; Mitrelias, T.; Kopper, K. P.; Steinmuller, S. J.; Bland, J. A. C.

2008-06-01

We report a new magnetic labelling technology for high-throughput biomolecular identification and DNA sequencing. Planar multi-bit magnetic tags have been designed and fabricated, which comprise a magnetic barcode formed by an ensemble of micron-sized thin film Ni80Fe20 bars encapsulated in SU8. We show that by using a globally applied magnetic field and magneto-optical Kerr microscopy the magnetic elements in the multi-bit magnetic tags can be addressed individually and encoded/decoded remotely. The critical steps needed to show the feasibility of this technology are demonstrated, including fabrication, flow transport, remote writing and reading, and successful functionalization of the tags as verified by fluorescence detection. This approach is ideal for encoding information on tags in microfluidic flow or suspension, for such applications as labelling of chemical precursors during drug synthesis and combinatorial library-based high-throughput multiplexed bioassays.

Hydroxyl Tagging Velocimetry in a Mach 2 Flow With a Wall Cavity (Postprint)

DTIC Science & Technology

2005-01-01

tagging velocimetry (HTV) measurements of velocity were made in a Mach 2 flow with a wall cavity. In the HTV method, ArF excimer laser (193 nm) beams...is tracked by planar laser -induced fluorescence. The grid motion over a fixed time delay yields about 50 velocity vectors of the two-dimensional flow...Mach 2 flow with a wall cavity. In the HTV method, ArF excimer laser (193 nm) beams pass through a humid gas and dissociate H2O into H + OH to form

High-Bandwidth Tactical-Network Data Analysis in a High-Performance-Computing (HPC) Environment: Time Tagging the Data

DTIC Science & Technology

2015-09-01

this report made use of posttest processing techniques to provide packet-level time tagging with an accuracy close to 3 µs relative to Coordinated...h set of test records. The process described herein made use of posttest processing techniques to provide packet-level time tagging with an accuracy

Studies of the Tumor Microenvironment in Pathogenesis of Neuroblastoma

DTIC Science & Technology

2012-07-01

DISTRIBUTION / AVAILABILITY STATEMENT 13. SUPPLEMENTARY NOTES 14. ABSTRACT The NBL -Tag neuroblastoma mice were crossed with B-cell... NBL -Tag mice were established. The NBL -Tag/B-cell deficient mice lacked B-cells as expected using flow cytometry analyses and immunohistochemistry...However, the lack of B-cells did not alter the growth patterns of NBL -Tag tumor formation as imaged by MRI. Studies using anti-B cell therapy were

Assessment Of Coronary Artery Aneurysms Using Transluminal Attenuation Gradient And Computational Modeling In Kawasaki Disease Patients

NASA Astrophysics Data System (ADS)

Grande Gutierrez, Noelia; Kahn, Andrew; Shirinsky, Olga; Gagarina, Nina; Lyskina, Galina; Fukazawa, Ryuji; Owaga, Shunichi; Burns, Jane; Marsden, Alison

2015-11-01

Kawasaki Disease (KD) can result in coronary artery aneurysms (CAA) in up to 25% of patients, putting them at risk of thrombus formation, myocardial infarction and sudden death. Clinical guidelines recommend CAA diameter >8 mm as the arbitrary criterion for initiating systemic anticoagulation. KD patient specific modeling and flow simulations suggest that hemodynamic data can predict regions at increased risk of thrombosis. Transluminal Attenuation Gradient (TAG) is determined from the change in radiological attenuation per vessel length and has been proposed as a non-invasive method for characterizing coronary stenosis from CT Angiography. We hypothesized that CAA abnormal flow could be quantified using TAG. We computed hemodynamics for patient specific coronary models using a stabilized finite element method, coupled numerically to a lumped parameter network to model the heart and vascular boundary conditions. TAG was quantified in the major coronary arteries. We compared TAG for aneurysmal and normal arteries and we analyzed TAG correlation with hemodynamic and geometrical parameters. Our results suggest that TAG may provide hemodynamic data not available from anatomy alone. TAG represents a possible extension to standard CTA that could help to better evaluate the risk of thrombus formation in KD.

Measurement of Lipid Accumulation in Chlorella vulgaris via Flow Cytometry and Liquid-State ¹H NMR Spectroscopy for Development of an NMR-Traceable Flow Cytometry Protocol

PubMed Central

Bono Jr., Michael S.; Garcia, Ravi D.; Sri-Jayantha, Dylan V.; Ahner, Beth A.; Kirby, Brian J.

2015-01-01

In this study, we cultured Chlorella vulgaris cells with a range of lipid contents, induced via nitrogen starvation, and characterized them via flow cytometry, with BODIPY 505/515 as a fluorescent lipid label, and liquid-state 1H NMR spectroscopy. In doing so, we demonstrate the utility of calibrating flow cytometric measurements of algal lipid content using triacylglyceride (TAG, also known as triacylglycerol or triglyceride) content per cell as measured via quantitative 1H NMR. Ensemble-averaged fluorescence of BODIPY-labeled cells was highly correlated with average TAG content per cell measured by bulk NMR, with a linear regression yielding a linear fit with r 2 = 0.9974. This correlation compares favorably to previous calibrations of flow cytometry protocols to lipid content measured via extraction, and calibration by NMR avoids the time and complexity that is generally required for lipid quantitation via extraction. Flow cytometry calibrated to a direct measurement of TAG content can be used to investigate the distribution of lipid contents for cells within a culture. Our flow cytometry measurements showed that Chlorella vulgaris cells subjected to nitrogen limitation exhibited higher mean lipid content but a wider distribution of lipid content that overlapped the relatively narrow distribution of lipid content for replete cells, suggesting that nitrogen limitation induces lipid accumulation in only a subset of cells. Calibration of flow cytometry protocols using direct in situ measurement of TAG content via NMR will facilitate rapid development of more precise flow cytometry protocols, enabling investigation of algal lipid accumulation for development of more productive algal biofuel feedstocks and cultivation protocols. PMID:26267664

Measurement of lipid accumulation in Chlorella vulgaris via flow cytometry and liquid-state ¹H NMR spectroscopy for development of an NMR-traceable flow cytometry protocol.

PubMed

Bono, Michael S; Garcia, Ravi D; Sri-Jayantha, Dylan V; Ahner, Beth A; Kirby, Brian J

2015-01-01

In this study, we cultured Chlorella vulgaris cells with a range of lipid contents, induced via nitrogen starvation, and characterized them via flow cytometry, with BODIPY 505/515 as a fluorescent lipid label, and liquid-state 1H NMR spectroscopy. In doing so, we demonstrate the utility of calibrating flow cytometric measurements of algal lipid content using triacylglyceride (TAG, also known as triacylglycerol or triglyceride) content per cell as measured via quantitative 1H NMR. Ensemble-averaged fluorescence of BODIPY-labeled cells was highly correlated with average TAG content per cell measured by bulk NMR, with a linear regression yielding a linear fit with r2 = 0.9974. This correlation compares favorably to previous calibrations of flow cytometry protocols to lipid content measured via extraction, and calibration by NMR avoids the time and complexity that is generally required for lipid quantitation via extraction. Flow cytometry calibrated to a direct measurement of TAG content can be used to investigate the distribution of lipid contents for cells within a culture. Our flow cytometry measurements showed that Chlorella vulgaris cells subjected to nitrogen limitation exhibited higher mean lipid content but a wider distribution of lipid content that overlapped the relatively narrow distribution of lipid content for replete cells, suggesting that nitrogen limitation induces lipid accumulation in only a subset of cells. Calibration of flow cytometry protocols using direct in situ measurement of TAG content via NMR will facilitate rapid development of more precise flow cytometry protocols, enabling investigation of algal lipid accumulation for development of more productive algal biofuel feedstocks and cultivation protocols.

Communication methods, systems, apparatus, and devices involving RF tag registration

DOEpatents

Burghard, Brion J [W. Richland, WA; Skorpik, James R [Kennewick, WA

2008-04-22

One technique of the present invention includes a number of Radio Frequency (RF) tags that each have a different identifier. Information is broadcast to the tags from an RF tag interrogator. This information corresponds to a maximum quantity of tag response time slots that are available. This maximum quantity may be less than the total number of tags. The tags each select one of the time slots as a function of the information and a random number provided by each respective tag. The different identifiers are transmitted to the interrogator from at least a subset of the RF tags.

In situ tagging technique for fishes provides insight into growth and movement of invasive lionfish

PubMed Central

Akins, John L; Morris, James A; Green, Stephanie J

2014-01-01

Information on fish movement and growth is primarily obtained through the marking and tracking of individuals with external tags, which are usually affixed to anesthetized individuals at the surface. However, the quantity and quality of data obtained by this method is often limited by small sample sizes owing to the time associated with the tagging process, high rates of tagging-related mortality, and displacement of tagged individuals from the initial capture location. To address these issues, we describe a technique for applying external streamer and dart tags in situ, which uses SCUBA divers to capture and tag individual fish on the sea floor without the use of anesthetic. We demonstrate this method for Indo-Pacific lionfish (Pterois volitans/P. miles), species which are particularly vulnerable to barotrauma when transported to and handled at the surface. To test our method, we tagged 161 individuals inhabiting 26 coral reef locations in the Bahamas over a period of 3 years. Our method resulted in no instances of barotrauma, reduced handling and recovery time, and minimal post-tagging release displacement compared with conventional ex situ tag application. Opportunistic resighting and recapture of tagged individuals reveals that lionfish exhibit highly variable site fidelity, movement patterns, and growth rates on invaded coral reef habitats. In total, 24% of lionfish were resighted between 29 and 188 days after tagging. Of these, 90% were located at the site of capture, while the remaining individuals were resighted between 200 m and 1.1 km from initial site of capture over 29 days later. In situ growth rates ranged between 0.1 and 0.6 mm/day. While individuals tagged with streamer tags posted slower growth rates with increasing size, as expected, there was no relationship between growth rate and fish size for individuals marked with dart tags, potentially because of large effects of tag presence on the activities of small bodied lionfish (i.e., <150 mm), where the tag was up to 7.6% of the lionfish's mass. Our study offers a novel in situ tagging technique that can be used to provide critical information on fish site fidelity, movement patterns, and growth in cases where ex situ tagging is not feasible. PMID:25614791

In situ tagging technique for fishes provides insight into growth and movement of invasive lionfish.

PubMed

Akins, John L; Morris, James A; Green, Stephanie J

2014-10-01

Information on fish movement and growth is primarily obtained through the marking and tracking of individuals with external tags, which are usually affixed to anesthetized individuals at the surface. However, the quantity and quality of data obtained by this method is often limited by small sample sizes owing to the time associated with the tagging process, high rates of tagging-related mortality, and displacement of tagged individuals from the initial capture location. To address these issues, we describe a technique for applying external streamer and dart tags in situ, which uses SCUBA divers to capture and tag individual fish on the sea floor without the use of anesthetic. We demonstrate this method for Indo-Pacific lionfish (Pterois volitans/P. miles), species which are particularly vulnerable to barotrauma when transported to and handled at the surface. To test our method, we tagged 161 individuals inhabiting 26 coral reef locations in the Bahamas over a period of 3 years. Our method resulted in no instances of barotrauma, reduced handling and recovery time, and minimal post-tagging release displacement compared with conventional ex situ tag application. Opportunistic resighting and recapture of tagged individuals reveals that lionfish exhibit highly variable site fidelity, movement patterns, and growth rates on invaded coral reef habitats. In total, 24% of lionfish were resighted between 29 and 188 days after tagging. Of these, 90% were located at the site of capture, while the remaining individuals were resighted between 200 m and 1.1 km from initial site of capture over 29 days later. In situ growth rates ranged between 0.1 and 0.6 mm/day. While individuals tagged with streamer tags posted slower growth rates with increasing size, as expected, there was no relationship between growth rate and fish size for individuals marked with dart tags, potentially because of large effects of tag presence on the activities of small bodied lionfish (i.e., <150 mm), where the tag was up to 7.6% of the lionfish's mass. Our study offers a novel in situ tagging technique that can be used to provide critical information on fish site fidelity, movement patterns, and growth in cases where ex situ tagging is not feasible.

Behavior and movement of formerly landlocked juvenile coho salmon after release into the free-flowing Cowlitz River, Washington

USGS Publications Warehouse

Kock, Tobias J.; Henning, Julie A.; Liedtke, Theresa L.; Royer, Ida M.; Ekstrom, Brian K.; Rondorf, Dennis W.

2011-01-01

Formerly landlocked Coho Salmon (Oncorhynchus kisutch) juveniles (age 2) were monitored following release into the free-flowing Cowlitz River to determine if they remained in the river or resumed seaward migration. Juvenile Coho Salmon were tagged with a radio transmitter (30 fish) or Floy tag (1050 fish) and their behavior was monitored in the lower Cowlitz River. We found that 97% of the radio-tagged fish remained in the Cowlitz River beyond the juvenile outmigration period, and the number of fish dispersing downstream decreased with increasing distance from the release site. None of the tagged fish returned as spawning adults in the 2 y following release. We suspect that fish in our study failed to migrate because they exceeded a threshold in size, age, or physiological status. Tagged fish in our study primarily remained in the Cowlitz River, thus it is possible that these fish presented challenges to juvenile salmon migrating through the system either directly by predation or indirectly by competition for food or habitat. Given these findings, returning formerly landlocked Coho Salmon juveniles to the free-flowing river apparently provided no benefit to the anadromous population. These findings have management implications in locations where landlocked salmon have the potential to interact with anadromous species of concern.

Single cell analysis using surface enhanced Raman scattering (SERS) tags

PubMed Central

Nolan, John P.; Duggan, Erika; Liu, Er; Condello, Danilo; Dave, Isha; Stoner, Samuel A.

2013-01-01

Fluorescence is a mainstay of bioanalytical methods, offering sensitive and quantitative reporting, often in multiplexed or multiparameter assays. Perhaps the best example of the latter is flow cytometry, where instruments equipped with multiple lasers and detectors allow measurement of 15 or more different fluorophores simultaneously, but increases beyond this number are limited by the relatively broad emission spectra. Surface enhanced Raman scattering (SERS) from metal nanoparticles can produce signal intensities that rival fluorescence, but with narrower spectral features that allow a greater degree of multiplexing. We are developing nanoparticle SERS tags as well as Raman flow cytometers for multiparameter single cell analysis of suspension or adherent cells. SERS tags are based on plasmonically active nanoparticles (gold nanorods) whose plasmon resonance can be tuned to give optimal SERS signals at a desired excitation wavelength. Raman resonant compounds are adsorbed on the nanoparticles to confer a unique spectral fingerprint on each SERS tag, which are then encapsulated in a polymer coating for conjugation to antibodies or other targeting molecules. Raman flow cytometry employs a high resolution spectral flow cytometer capable of measuring the complete SERS spectra, as well as conventional flow cytometry measurements, from thousands of individual cells per minute. Automated spectral unmixing algorithms extract the contributions of each SERS tag from each cell to generate high content, multiparameter single cell population data. SERS-based cytometry is a powerful complement to conventional fluorescence-based cytometry. The narrow spectral features of the SERS signal enables more distinct probes to be measured in a smaller region of the optical spectrum with a single laser and detector, allowing for higher levels of multiplexing and multiparameter analysis. PMID:22498143

Direct Measurement of Lung Motion Using Hyperpolarized Helium-3 MR Tagging

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

Cai Jing; Miller, G. Wilson; Altes, Talissa A.

2007-07-01

Purpose: To measure lung motion between end-inhalation and end-exhalation using a hyperpolarized helium-3 (HP {sup 3}He) magnetic resonance (MR) tagging technique. Methods and Materials: Three healthy volunteers underwent MR tagging studies after inhalation of 1 L HP {sup 3}He gas diluted with nitrogen. Multiple-slice two-dimensional and volumetric three-dimensional MR tagged images of the lungs were obtained at end-inhalation and end-exhalation, and displacement vector maps were computed. Results: The grids of tag lines in the HP {sup 3}He MR images were well defined at end-inhalation and remained evident at end-exhalation. Displacement vector maps clearly demonstrated the regional lung motion and deformationmore » that occurred during exhalation. Discontinuity and differences in motion pattern between two adjacent lung lobes were readily resolved. Conclusions: Hyperpolarized helium-3 MR tagging technique can be used for direct in vivo measurement of respiratory lung motion on a regional basis. This technique may lend new insights into the regional pulmonary biomechanics and thus provide valuable information for the deformable registration of lung.« less

Development and use of in-stream PIT-tag detection systems to assess movement behavior of fish in tributaries of the Columbia River Basin, USA

USGS Publications Warehouse

Connolly, P.J.; Jezorek, I.G.; Prentice, E.F.

2005-01-01

We have developed detector systems for fish implanted with Passive Integrated Transponder (PIT) tags to assess their movement behavior and habitat use within fast flowing streams. Fish tested have primarily been wild anadromous and resident forms of rainbow trout Oncorhynchus mykiss and cutthroat trout O. clarki. Longitudinal arrangements of two- and six-antennas allow determination of direction of movement and efficiency of detection. Our first detector system became operational in August 2001, with subsequent improvements over time. In tests with a two-antenna system, detection efficiency of tagged, downstreammoving fish was high (96%) during low flows, but less (69%) during high flows. With an increase in the number of antennas to six, arranged in a 2x3 array, the detection efficiency of downstream-moving fish was increased to 95-100% at all flows. Detection efficiency of upstream-moving fish was high (95-100%) in both the two-and six-antenna system during all flows. Antennas were anchored to the substrate and largely spanned the bank-full width. Modifications to the methods used to anchor antennas have increased the likelihood of the system remaining intact and running at full detection capability during challenging flow and debris conditions, largely achieving our goal to have continuous monitoring of fish movement throughout an annual cycle. In August 2004, we placed a similar detector system in another watershed. Success has much relied on the quality of transceivers and electrical power. Detection of tagged fish passing our static PIT-tag detectors has produced valuable information on how selected fish species use the network of streams in a watershed. Integrating information from our detectors in tributary streams with that from detectors downstream at dams in the Columbia River has promise to be a powerful tool for monitoring movement patterns of anadromous fish species and to understanding full lifecycle fish behavior and habitat use.

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

Swansiger, W.A.; Shepodd, T.J.; Phillips, M.L.F.

The ability to identify the manufacturers and distributors of chemicals seized in raids of illicit drug labs would be of great value in controlling the diversion of these chemicals. We developed a tagging scheme based on the addition of sub-ppM concentrations of various combinations of rare-earth elements to the target chemicals and evaluated a number of techniques for detecting the tags. We developed soluble tags for tagging liquids and selected Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) as the preferred detection technique. We developed insoluble tags for tagging solids and developed methods to analyze them and mix them into solid precursors. Wemore » have successfully demonstrated the tagging of several solvents and two of the precursor chemicals used in one of the most popular clandestine methamphetamine syntheses (ephedrine reacting with hydriodic acid/red phosphorus). The tagging scheme is capable of yielding tens of thousands of signatures (using holmium as an internal standard and up to 9 rare-earths at up to 3 concentrations yields 3{sup 9} {minus} 1 = 19,682 signatures) and is applicable to most of the chemicals on the precursor and essential chemicals list. In the concentrations employed, the tags are safe enough to be added to pharmaceuticals and cheap enough to tag tanker loads of chemicals.« less

Quartz Crystal Microbalance with Dissipation Monitoring

DTIC Science & Technology

2014-11-06

Hydroxyapatite , 10 nm, Sensors • Biotin Functionalized on Gold Sensors • His-tag Capturing Sensor QCM-D techniques provide answers about...UV/Ozone cleaner • Hydroxyapatite , 10 nm, Sensors • Biotin Functionalized on Gold Sensors • His-tag Capturing Sensor QCM-D techniques provide

Informatics in Radiology: Dual-Energy Electronic Cleansing for Fecal-Tagging CT Colonography

PubMed Central

Kim, Se Hyung; Lee, June-Goo; Yoshida, Hiroyuki

2013-01-01

Electronic cleansing (EC) is an emerging technique for the removal of tagged fecal materials at fecal-tagging computed tomographic (CT) colonography. However, existing EC methods may generate various types of artifacts that severely impair the quality of the cleansed CT colonographic images. Dual-energy fecal-tagging CT colonography is regarded as a next-generation imaging modality. EC that makes use of dual-energy fecal-tagging CT colonographic images promises to be effective in reducing cleansing artifacts by means of applying the material decomposition capability of dual-energy CT. The dual-energy index (DEI), which is calculated from the relative change in the attenuation values of a material at two different photon energies, is a reliable and effective indicator for differentiating tagged fecal materials from various types of tissues on fecal-tagging CT colonographic images. A DEI-based dual-energy EC scheme uses the DEI to help differentiate the colonic lumen—including the luminal air, tagged fecal materials, and air-tagging mixture—from the colonic soft-tissue structures, and then segments the entire colonic lumen for cleansing of the tagged fecal materials. As a result, dual-energy EC can help identify partial-volume effects in the air-tagging mixture and inhomogeneous tagging in residual fecal materials, the major causes of EC artifacts. This technique has the potential to significantly improve the quality of EC and promises to provide images of a cleansed colon that are free of the artifacts commonly observed with conventional single-energy EC methods. © RSNA, 2013 PMID:23479680

An optimized design to reduce eddy current sensitivity in velocity-selective arterial spin labeling using symmetric BIR-8 pulses.

PubMed

Guo, Jia; Meakin, James A; Jezzard, Peter; Wong, Eric C

2015-03-01

Velocity-selective arterial spin labeling (VSASL) tags arterial blood on a velocity-selective (VS) basis and eliminates the tagging/imaging gap and associated transit delay sensitivity observed in other ASL tagging methods. However, the flow-weighting gradient pulses in VS tag preparation can generate eddy currents (ECs), which may erroneously tag the static tissue and create artificial perfusion signal, compromising the accuracy of perfusion quantification. A novel VS preparation design is presented using an eight-segment B1 insensitive rotation with symmetric radio frequency and gradient layouts (sym-BIR-8), combined with delays after gradient pulses to optimally reduce ECs of a wide range of time constants while maintaining B0 and B1 insensitivity. Bloch simulation, phantom, and in vivo experiments were carried out to determine robustness of the new and existing pulse designs to ECs, B0 , and B1 inhomogeneity. VSASL with reduced EC sensitivity across a wide range of EC time constants was achieved with the proposed sym-BIR-8 design, and the accuracy of cerebral blood flow measurement was improved. The sym-BIR-8 design performed the most robustly among the existing VS tagging designs, and should benefit studies using VS preparation with improved accuracy and reliability. © 2014 Wiley Periodicals, Inc.

Contribution of lateral terrestrial water flows to the regional hydrological cycle: A joint soil-atmospheric moisture tagging procedure with WRF-Hydro

NASA Astrophysics Data System (ADS)

Arnault, Joel; Wei, Jianhui; Zhang, Zhenyu; Wagner, Sven; Kunstmann, Harald

2017-04-01

Water resources management requires an accurate knowledge of the behavior of the regional hydrological cycle components, including precipitation, evapotranspiration, river discharge and soil water storage. Atmospheric models such as the Weather Research and Forecasting (WRF) model provide a tool to evaluate these components. The main drawback of these atmospheric models, however, is that the terrestrial segment of the hydrological cycle is reduced to vertical infiltration, and that lateral terrestrial water flows are neglected. Recent model developments have focused on coupled atmospheric-hydrological modeling systems, such as WRF-hydro, in order to take into account subsurface, overland and river flow. The aim of this study is to investigate the contribution of lateral terrestrial water flows to the regional hydrological cycle, with the help of a joint soil-atmospheric moisture tagging procedure. This procedure is the extended version of an existing atmospheric moisture tagging method developed in WRF and WRF-Hydro (Arnault et al. 2017). It is used to quantify the partitioning of precipitation into water stored in the soil, runoff, evapotranspiration, and potentially subsequent precipitation through regional recycling. An application to a high precipitation event on 23 June 2009 in the upper Danube river basin, Germany and Austria, is presented. Precipitating water during this day is tagged for the period 2009-2011. Its contribution to runoff and evapotranspiration decreases with time, but is still not negligible in the summer 2011. At the end of the study period, less than 5 % of the precipitating water on 23 June 2009 remains in the soil. The additionally resolved lateral terrestrial water flows in WRF-Hydro modify the partitioning between surface and underground runoff, in association with a slight increase of evapotranspiration and recycled precipitation. Reference: Arnault, J., R. Knoche, J. Wei, and H. Kunstmann (2016), Evaporation tagging and atmospheric water budget analysis with WRF: A regional precipitation recycling study for West Africa, Water Resour. Res., 52, 1544-1567, doi:10.1002/2015WR017704.

Genetically encoded fluorescent tags

PubMed Central

Thorn, Kurt

2017-01-01

Genetically encoded fluorescent tags are protein sequences that can be fused to a protein of interest to render it fluorescent. These tags have revolutionized cell biology by allowing nearly any protein to be imaged by light microscopy at submicrometer spatial resolution and subsecond time resolution in a live cell or organism. They can also be used to measure protein abundance in thousands to millions of cells using flow cytometry. Here I provide an introduction to the different genetic tags available, including both intrinsically fluorescent proteins and proteins that derive their fluorescence from binding of either endogenous or exogenous fluorophores. I discuss their optical and biological properties and guidelines for choosing appropriate tags for an experiment. Tools for tagging nucleic acid sequences and reporter molecules that detect the presence of different biomolecules are also briefly discussed. PMID:28360214

Monitoring the Migrations of Wild Snake River Spring/Summer Chinook Salmon Juveniles, 2007-2008

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

Achord, Stephen; Sandford, Benjamin P.; Hockersmith, Eric E.

2009-07-09

This report provides results from an ongoing project to monitor the migration behavior and survival of wild juvenile spring/summer Chinook salmon in the Snake River Basin. Data reported is from detections of PIT tagged fish during late summer 2007 through mid-2008. Fish were tagged in summer 2007 by the National Marine Fisheries Service (NMFS) in Idaho and by the Oregon Department of Fish and Wildlife (ODFW) in Oregon. Our analyses include migration behavior and estimated survival of fish at instream PIT-tag monitors and arrival timing and estimated survival to Lower Granite Dam. Principal results from tagging and interrogation during 2007-2008more » are: (1) In July and August 2007, we PIT tagged and released 7,390 wild Chinook salmon parr in 12 Idaho streams or sample areas. (2) Overall observed mortality from collection, handling, tagging, and after a 24-hour holding period was 1.4%. (3) Of the 2,524 Chinook salmon parr PIT tagged and released in Valley Creek in summer 2007, 218 (8.6%) were detected at two instream PIT-tag monitoring systems in lower Valley Creek from late summer 2007 to the following spring 2008. Of these, 71.6% were detected in late summer/fall, 11.9% in winter, and 16.5% in spring. Estimated parr-to-smolt survival to Lower Granite Dam was 15.5% for the late summer/fall group, 48.0% for the winter group, and 58.5% for the spring group. Based on detections at downstream dams, the overall efficiency of VC1 (upper) or VC2 (lower) Valley Creek monitors for detecting these fish was 21.1%. Using this VC1 or VC2 efficiency, an estimated 40.8% of all summer-tagged parr survived to move out of Valley Creek, and their estimated survival from that point to Lower Granite Dam was 26.5%. Overall estimated parr-to-smolt survival for all summer-tagged parr from this stream at the dam was 12.1%. Development and improvement of instream PIT-tag monitoring systems continued throughout 2007 and 2008. (4) Testing of PIT-tag antennas in lower Big Creek during 2007-2008 showed these antennas (and anchoring method) are not adequate to withstand high spring flows in this drainage. Future plans involve removing these antennas before high spring flows. (5) At Little Goose Dam in 2008, length and/or weight were taken on 505 recaptured fish from 12 Idaho stream populations. Fish had grown an average of 40.1 mm in length and 10.6 g in weight over an average of 288 d. Their mean condition factor declined from 1.25 at release (parr) to 1.05 at recapture (smolt). (6) Mean release lengths for detected fish were significantly larger than for fish not detected the following spring and summer (P < 0.0001). (7) Fish that migrated through Lower Granite Dam in April and May were significantly larger at release than fish that migrated after May (P < 0.0001) (only 12 fish migrated after May). (8) In 2008, peak detections at Lower Granite Dam of parr tagged during summer 2007 (from the 12 stream populations in Idaho and 4 streams in Oregon) occurred during moderate flows of 87.5 kcfs on 7 May and high flows of 197.3 kcfs on 20 May. The 10th, 50th, and 90th percentile passage occurred on 30 April, 11 May, and 23 May, respectively. (9) In 2007-2008, estimated parr-to-smolt survival to Lower Granite Dam for Idaho and Oregon streams (combined) averaged 19.4% (range 6.2-38.4% depending on stream of origin). In Idaho streams the estimated parr-to-smolt survival averaged 21.0%. This survival was the second highest since 1993 for Idaho streams. Relative parr densities were lower in 2007 (2.4 parr/100 m2) than in all previous years since 2000. In 2008, we observed low-to-moderate flows prior to mid-May and relatively cold weather conditions throughout the spring migration season. These conditions moved half of the fish through Lower Granite Dam prior to mid-May; then high flows moved 50 to 90% of the fish through the dam in only 12 days. Clearly, complex interrelationships of several factors drive the annual migrational timing of the stocks.« less

Strain analysis in CRT candidates using the novel segment length in cine (SLICE) post-processing technique on standard CMR cine images.

PubMed

Zweerink, Alwin; Allaart, Cornelis P; Kuijer, Joost P A; Wu, LiNa; Beek, Aernout M; van de Ven, Peter M; Meine, Mathias; Croisille, Pierre; Clarysse, Patrick; van Rossum, Albert C; Nijveldt, Robin

2017-12-01

Although myocardial strain analysis is a potential tool to improve patient selection for cardiac resynchronization therapy (CRT), there is currently no validated clinical approach to derive segmental strains. We evaluated the novel segment length in cine (SLICE) technique to derive segmental strains from standard cardiovascular MR (CMR) cine images in CRT candidates. Twenty-seven patients with left bundle branch block underwent CMR examination including cine imaging and myocardial tagging (CMR-TAG). SLICE was performed by measuring segment length between anatomical landmarks throughout all phases on short-axis cines. This measure of frame-to-frame segment length change was compared to CMR-TAG circumferential strain measurements. Subsequently, conventional markers of CRT response were calculated. Segmental strains showed good to excellent agreement between SLICE and CMR-TAG (septum strain, intraclass correlation coefficient (ICC) 0.76; lateral wall strain, ICC 0.66). Conventional markers of CRT response also showed close agreement between both methods (ICC 0.61-0.78). Reproducibility of SLICE was excellent for intra-observer testing (all ICC ≥0.76) and good for interobserver testing (all ICC ≥0.61). The novel SLICE post-processing technique on standard CMR cine images offers both accurate and robust segmental strain measures compared to the 'gold standard' CMR-TAG technique, and has the advantage of being widely available. • Myocardial strain analysis could potentially improve patient selection for CRT. • Currently a well validated clinical approach to derive segmental strains is lacking. • The novel SLICE technique derives segmental strains from standard CMR cine images. • SLICE-derived strain markers of CRT response showed close agreement with CMR-TAG. • Future studies will focus on the prognostic value of SLICE in CRT candidates.

Particle Image Velocimetry Applications Using Fluorescent Dye-Doped Particles

NASA Technical Reports Server (NTRS)

Petrosky, Brian J.; Maisto, Pietro; Lowe, K. Todd; Andre, Matthieu A.; Bardet, Philippe M.; Tiemsin, Patsy I.; Wohl, Christopher J.; Danehy, Paul M.

2015-01-01

Polystyrene latex sphere particles are widely used to seed flows for velocimetry techniques such as Particle Image Velocimetry (PIV) and Laser Doppler Velocimetry (LDV). These particles may be doped with fluorescent dyes such that signals spectrally shifted from the incident laser wavelength may be detected via Laser Induced Fluorescence (LIF). An attractive application of the LIF signal is achieving velocimetry in the presence of strong interference from laser scatter, opening up new research possibilities very near solid surfaces or at liquid/gas interfaces. Additionally, LIF signals can be used to tag different fluid streams to study mixing. While fluorescence-based PIV has been performed by many researchers for particles dispersed in water flows, the current work is among the first in applying the technique to micron-scale particles dispersed in a gas. A key requirement for such an application is addressing potential health hazards from fluorescent dyes; successful doping of Kiton Red 620 (KR620) has enabled the use of this relatively safe dye for fluorescence PIV for the first time. In this paper, basic applications proving the concept of PIV using the LIF signal from KR620-doped particles are exhibited for a free jet and a twophase flow apparatus. Results indicate that while the fluorescence PIV techniques are roughly 2 orders of magnitude weaker than Mie scattering, they provide a viable method for obtaining data in flow regions previously inaccessible via standard PIV. These techniques have the potential to also complement Mie scattering signals, for example in multi-stream and/or multi-phase experiments.

Sensitive Carbohydrate Detection using Surface Enhanced Raman Tagging

PubMed Central

Vangala, Karthikeshwar; Yanney, Michael; Hsiao, Cheng-Te; Wu, Wells W.; Shen, Rong-Fong; Zou, Sige; Sygula, Andrzej; Zhang, Dongmao

2010-01-01

Glycomic analysis is an increasingly important field in biological and biomedical research as glycosylation is one of the most important protein post-translational modifications. We have developed a new technique to detect carbohydrates using surface enhanced Raman spectroscopy (SERS) by designing and applying a Rhodamine B derivative as the SERS tag. Using a reductive amination reaction, the Rhodamine-based tag (RT) was successfully conjugated to three model carbohydrates (glucose, lactose and glucuronic acid). SERS detection limits obtained with 632 nm HeNe laser were ~1 nM in concentration for all the RT-carbohydrate conjugates and ~10 fmol in total sample consumption. The dynamic range of the SERS method is about 4 orders of magnitude, spanning from 1 nM to 5 µM. Ratiometric SERS quantification using isotope-substituted SERS internal references also allows comparative quantifications of carbohydrates labeled with RT and deuterium/hydrogen substituted RT tags, respectively. In addition to enhancing the SERS detection of the tagged carbohydrates, the Rhodamine tagging facilitates fluorescence and mass spectrometric detection of carbohydrates. Current fluorescence sensitivity of RT-carbohydrates is ~ 3 nM in concentration while the mass spectrometry (MS) sensitivity is about 1 fmol that was achieved with linear ion trap electrospray ionization (ESI)-MS instrument. Potential applications that take advantage of the high SERS, fluorescence and MS sensitivity of this SERS tagging strategy are discussed for practical glycomic analysis where carbohydrates may be quantified with a fluorescence and SERS technique, and then identified with ESI-MS techniques. PMID:21082777

Array processing for RFID tag localization exploiting multi-frequency signals

NASA Astrophysics Data System (ADS)

Zhang, Yimin; Li, Xin; Amin, Moeness G.

2009-05-01

RFID is an increasingly valuable business and technology tool for electronically identifying, locating, and tracking products, assets, and personnel. As a result, precise positioning and tracking of RFID tags and readers have received considerable attention from both academic and industrial communities. Finding the position of RFID tags is considered an important task in various real-time locating systems (RTLS). As such, numerous RFID localization products have been developed for various applications. The majority of RFID positioning systems is based on the fusion of pieces of relevant information, such as the range and the direction-of-arrival (DOA). For example, trilateration can determine the tag position by using the range information of the tag estimated from three or more spatially separated reader antennas. Triangulation is another method to locate RFID tags that use the direction-of-arrival (DOA) information estimated at multiple spatially separated locations. The RFID tag positions can also be determined through hybrid techniques that combine the range and DOA information. The focus of this paper to study the design and performance of the localization of passive RFID tags using array processing techniques in a multipath environment, and exploiting multi-frequency CW signals. The latter are used to decorrelate the coherent multipath signals for effective DOA estimation and for the purpose of accurate range estimation. Accordingly, the spatial and frequency dimensionalities are fully utilized for robust and accurate positioning of RFID tags.

A new charge-tagged proline-based organocatalyst for mechanistic studies using electrospray mass spectrometry

PubMed Central

Willms, J Alexander; Beel, Rita; Schmidt, Martin L; Mundt, Christian

2014-01-01

Summary A new 4-hydroxy-L-proline derivative with a charged 1-ethylpyridinium-4-phenoxy substituent has been synthesized with the aim of facilitating mechanistic studies of proline-catalyzed reactions by ESI mass spectrometry. The charged residue ensures a strongly enhanced ESI response compared to neutral unmodified proline. The connection by a rigid linker fixes the position of the charge tag far away from the catalytic center in order to avoid unwanted interactions. The use of a charged catalyst leads to significantly enhanced ESI signal abundances for every catalyst-derived species which are the ones of highest interest present in a reacting solution. The new charged proline catalyst has been tested in the direct asymmetric inverse aldol reaction between aldehydes and diethyl ketomalonate. Two intermediates in accordance with the List–Houk mechanism for enamine catalysis have been detected and characterized by gas-phase fragmentation. In addition, their temporal evolution has been followed using a microreactor continuous-flow technique. PMID:25246962

Effects of a spring flushing flow on the distribution of radio-tagged juvenile rainbow trout in a Wyoming tailwater

USGS Publications Warehouse

Simpkins, D.G.; Hubert, W.A.; Wesche, T.A.

2000-01-01

The controlled release of dammed water, designed to produce a flushing flow that would remove fine sediments from spawning habitat in a flow-regulated river, did not displace juvenile rainbow trout Oncorhynchus mykiss (20-25 cm total length) downstream. Of eight naturally spawned (wild) and nine hatchery fish that were radio-tagged, only one wild fish and two hatchery fish were found in different locations after an eight-fold increase in flow. These three fish moved upstream and the greatest movement was 96 m. Cobble and boulder substrates in main-channel pools were thought to provide slow-water-velocity refuges during the flushing flow. Our findings and the findings of other researchers suggest that flushing flows can be conducted to enhance spawning habitat for rainbow trout without causing extensive downstream movements or habitat displacement of small fish in regulated rivers.

Security techniques for prevention of rank manipulation in social tagging services including robotic domains.

PubMed

Choi, Okkyung; Jung, Hanyoung; Moon, Seungbin

2014-01-01

With smartphone distribution becoming common and robotic applications on the rise, social tagging services for various applications including robotic domains have advanced significantly. Though social tagging plays an important role when users are finding the exact information through web search, reliability and semantic relation between web contents and tags are not considered. Spams are making ill use of this aspect and put irrelevant tags deliberately on contents and induce users to advertise contents when they click items of search results. Therefore, this study proposes a detection method for tag-ranking manipulation to solve the problem of the existing methods which cannot guarantee the reliability of tagging. Similarity is measured for ranking the grade of registered tag on the contents, and weighted values of each tag are measured by means of synonym relevance, frequency, and semantic distances between tags. Lastly, experimental evaluation results are provided and its efficiency and accuracy are verified through them.

Little Goose Dam Full Flow PIT-Tag Detection System Project Summary.

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

Warf, Don; Livingston, Scott

2009-04-16

In 2006, the design phase of this project was kicked off and was for the most part modeled after the Full Flow PIT installation installed at Lower Monumental Dam during winter and spring of 2006 and 2007. As the Goose Full Flow design progressed and the project started to move towards construction, issues within contracting occurred and the project was put on delay for 1 year. Starting in mid December of 2008, Harcon Inc. was awarded the contract and construction of the new Goose Full Flow PIT-tag detection system began. The purpose of this document is to summarize the installationmore » of the Little Goose Full Flow project from start to finish and to highlight the notable successes and challenges that the installation presented along with the final results and current status.« less

MO-G-18C-03: Evaluation of Deformable Image Registration for Lung Motion Estimation Using Hyperpolarized Gas Tagging MRI

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

Huang, Q; Zhang, Y; Liu, Y

2014-06-15

Purpose: Hyperpolarized gas (HP) tagging MRI is a novel imaging technique for direct measurement of lung motion during breathing. This study aims to quantitatively evaluate the accuracy of deformable image registration (DIR) in lung motion estimation using HP tagging MRI as references. Methods: Three healthy subjects were imaged using the HP MR tagging, as well as a high-resolution 3D proton MR sequence (TrueFISP) at the end-of-inhalation (EOI) and the end-of-exhalation (EOE). Ground truth of lung motion and corresponding displacement vector field (tDVF) was derived from HP tagging MRI by manually tracking the displacement of tagging grids between EOI and EOE.more » Seven different DIR methods were applied to the high-resolution TrueFISP MR images (EOI and EOE) to generate the DIR-based DVFs (dDVF). The DIR methods include Velocity (VEL), MIM, Mirada, multi-grid B-spline from Elastix (MGB) and 3 other algorithms from DIRART toolbox (Double Force Demons (DFD), Improved Lucas-Kanade (ILK), and Iterative Optical Flow (IOF)). All registrations were performed by independent experts. Target registration error (TRE) was calculated as tDVF – dDVF. Analysis was performed for the entire lungs, and separately for the upper and lower lungs. Results: Significant differences between tDVF and dDVF were observed. Besides the DFD and IOF algorithms, all other dDVFs showed similarity in deformation magnitude distribution but away from the ground truth. The average TRE for entire lung ranged 2.5−23.7mm (mean=8.8mm), depending on the DIR method and subject's breathing amplitude. Larger TRE (13.3–23.7mm) was found in subject with larger breathing amplitude of 45.6mm. TRE was greater in lower lung (2.5−33.9 mm, mean=12.4mm) than that in upper lung (2.5−11.9 mm, mean=5.8mm). Conclusion: Significant differences were observed in lung motion estimation between the HP gas tagging MRI method and the DIR methods, especially when lung motion is large. Large variation among different DIR methods was also observed.« less

Authentication via wavefront-shaped optical responses

NASA Astrophysics Data System (ADS)

Eilers, Hergen; Anderson, Benjamin R.; Gunawidjaja, Ray

2018-02-01

Authentication/tamper-indication is required in a wide range of applications, including nuclear materials management and product counterfeit detection. State-of-the-art techniques include reflective particle tags, laser speckle authentication, and birefringent seals. Each of these passive techniques has its own advantages and disadvantages, including the need for complex image comparisons, limited flexibility, sensitivity to environmental conditions, limited functionality, etc. We have developed a new active approach to address some of these short-comings. The use of an active characterization technique adds more flexibility and additional layers of security over current techniques. Our approach uses randomly-distributed nanoparticles embedded in a polymer matrix (tag/seal) which is attached to the item to be secured. A spatial light modulator is used to adjust the wavefront of a laser which interacts with the tag/seal, and a detector is used to monitor this interaction. The interaction can occur in various ways, including transmittance, reflectance, fluorescence, random lasing, etc. For example, at the time of origination, the wavefront-shaped reflectance from a tag/seal can be adjusted to result in a specific pattern (symbol, words, etc.) Any tampering with the tag/seal would results in a disturbance of the random orientation of the nanoparticles and thus distort the reflectance pattern. A holographic waveplate could be inserted into the laser beam for verification. The absence/distortion of the original pattern would then indicate that tampering has occurred. We have tested the tag/seal's and authentication method's tamper-indicating ability using various attack methods, including mechanical, thermal, and chemical attacks, and have verified our material/method's robust tamper-indicating ability.

An orange fluorescent protein tagging system for real-time pollen tracking.

PubMed

Rice, J Hollis; Millwood, Reginald J; Mundell, Richard E; Chambers, Orlando D; Abercrombie, Laura L; Davies, H Maelor; Stewart, C Neal

2013-09-27

Monitoring gene flow could be important for future transgenic crops, such as those producing plant-made-pharmaceuticals (PMPs) in open field production. A Nicotiana hybrid (Nicotiana. tabacum × Nicotiana glauca) shows limited male fertility and could be used as a bioconfined PMP platform. Effective assessment of gene flow from these plants is augmented with methods that utilize fluorescent proteins for transgenic pollen identification. We report the generation of a pollen tagging system utilizing an orange fluorescent protein to monitor pollen flow and as a visual assessment of transgene zygosity of the parent plant. This system was created to generate a tagged Nicotiana hybrid that could be used for the incidence of gene flow. Nicotiana tabacum 'TN 90' and Nicotiana glauca were successfully transformed via Agrobacterium tumefaciens to express the orange fluorescent protein gene, tdTomato-ER, in pollen and a green fluorescent protein gene, mgfp5-er, was expressed in vegetative structures of the plant. Hybrids were created that utilized the fluorescent proteins as a research tool for monitoring pollen movement and gene flow. Manual greenhouse crosses were used to assess hybrid sexual compatibility with N. tabacum, resulting in seed formation from hybrid pollination in 2% of crosses, which yielded non-viable seed. Pollen transfer to the hybrid formed seed in 19% of crosses and 10 out of 12 viable progeny showed GFP expression. The orange fluorescent protein is visible when expressed in the pollen of N. glauca, N. tabacum, and the Nicotiana hybrid, although hybrid pollen did not appear as bright as the parent lines. The hybrid plants, which show limited ability to outcross, could provide bioconfinement with the benefit of detectable pollen using this system. Fluorescent protein-tagging could be a valuable tool for breeding and in vivo ecological monitoring.

A scalable correlator for multichannel diffuse correlation spectroscopy.

PubMed

Stapels, Christopher J; Kolodziejski, Noah J; McAdams, Daniel; Podolsky, Matthew J; Fernandez, Daniel E; Farkas, Dana; Christian, James F

2016-02-01

Diffuse correlation spectroscopy (DCS) is a technique which enables powerful and robust non-invasive optical studies of tissue micro-circulation and vascular blood flow. The technique amounts to autocorrelation analysis of coherent photons after their migration through moving scatterers and subsequent collection by single-mode optical fibers. A primary cost driver of DCS instruments are the commercial hardware-based correlators, limiting the proliferation of multi-channel instruments for validation of perfusion analysis as a clinical diagnostic metric. We present the development of a low-cost scalable correlator enabled by microchip-based time-tagging, and a software-based multi-tau data analysis method. We will discuss the capabilities of the instrument as well as the implementation and validation of 2- and 8-channel systems built for live animal and pre-clinical settings.

Noninvasive physiologic assessment of coronary stenoses using cardiac CT.

PubMed

Xu, Lei; Sun, Zhonghua; Fan, Zhanming

2015-01-01

Coronary CT angiography (CCTA) has become an important noninvasive imaging modality in the diagnosis of coronary artery disease (CAD). CCTA enables accurate evaluation of coronary artery stenosis. However, CCTA provides limited information on the physiological significance of stenotic lesions. A noninvasive "one-stop-shop" diagnostic test that can provide both anatomical significance and functional significance of stenotic lesions would be beneficial in the diagnosis and management of CAD. Recently, with the introduction of novel techniques, such as myocardial CT perfusion, CT-derived fractional flow reserve (FFRCT), and transluminal attenuation gradient (TAG), CCTA has emerged as a noninvasive method for the assessment of both anatomy of coronary lesions and its physiological consequences during a single study. This review provides an overview of the current status of new CT techniques for the physiologic assessments of CAD.

Strep-Tagged Protein Purification.

PubMed

Maertens, Barbara; Spriestersbach, Anne; Kubicek, Jan; Schäfer, Frank

2015-01-01

The Strep-tag system can be used to purify recombinant proteins from any expression system. Here, protocols for lysis and affinity purification of Strep-tagged proteins from E. coli, baculovirus-infected insect cells, and transfected mammalian cells are given. Depending on the amount of Strep-tagged protein in the lysate, a protocol for batch binding and subsequent washing and eluting by gravity flow can be used. Agarose-based matrices with the coupled Strep-Tactin ligand are the resins of choice, with a binding capacity of up to 9 mg ml(-1). For purification of lower amounts of Strep-tagged proteins, the use of Strep-Tactin magnetic beads is suitable. In addition, Strep-tagged protein purification can also be automated using prepacked columns for FPLC or other liquid-handling chromatography instrumentation, but automated purification is not discussed in this protocol. The protocols described here can be regarded as an update of the Strep-Tag Protein Handbook (Qiagen, 2009). © 2015 Elsevier Inc. All rights reserved.

Quantitative NO{sub 2} molecular tagging velocimetry at 500 kHz frame rate

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

Jiang, Naibo; Nishihara, Munetake; Lempert, Walter R.

2010-11-29

NO{sub 2} molecular tagging velocimetry (MTV) is demonstrated at repetition rates as high as 500 kHz in a laboratory scale Mach 5 wind tunnel. A pulse burst laser and a home built optical parametric oscillator system were used to simultaneously generate the required 355 and 226 nm wavelengths for NO{sub 2} photodissociation (tagging) and NO planar laser induced fluorescence imaging (interrogation), respectively. NO{sub 2} MTV images were obtained both in front and behind the characteristic bow shock from a 5 mm diameter cylinder. From Gaussian curve fitting, an average free stream flow velocity of 719 m/s was obtained. Absolute statisticalmore » precision in velocity of {approx}11.5 m/s was determined, corresponding to relative precision of 1.6%-5%, depending upon the region of the flow probed.« less

Behavior and movement of adult summer steelhead following collection and release, lower Cowlitz River, Washington, 2012--2013

USGS Publications Warehouse

Kock, Tobias J.; Liedtke, Theresa L.; Ekstrom, Brian K.; Rondorf, Dennis W.; Gleizes, Chris; Dammers, Wolf; Gibson, Scott; Murphy, Jamie

2013-01-01

Executive SummaryHistorically, adult summer steelhead Oncorhynchus mykiss returning to hatcheries on the lower Cowlitz River were sometimes transported and released in the river (recycled) to provide additional angling opportunity for the popular sport fishery in the basin. However, this practice has not been used in recent years because of concerns associated with interactions between hatchery fish and wild fish. Fishery managers were interested in resuming recycling but lacked information regarding effects of this practice on wild steelhead so we conducted a study during 2012–2013 to: (1) enumerate recycled steelhead that returned to the hatchery or were removed by anglers; and (2) determine if steelhead that were not removed from the river remained in the system where they could interact with wild fish.During June–August 2012, a total of 549 summer steelhead were captured at the Cowlitz Salmon Hatchery, tagged, and released downstream near the Interstate 5 Bridge. All recycled steelhead were tagged with a white Floy® tag and opercle-punched; 109 (20 percent) of these fish also were radio-tagged. All adult steelhead that return to the hatchery were handled by hatchery staff so recycled steelhead that returned to the hatchery were enumerated daily. A creel survey and voluntary angler reports were used to determine the number of recycled steelhead that were caught by anglers. We established three fixed telemetry monitoring sites on the mainstem Cowlitz River and eight additional sites were deployed on tributaries to the lower Cowlitz River where wild winter steelhead are known to spawn. We also conducted mobile tracking from a boat during October 2012, November 2012, and January 2013 to locate radio-tagged fish.A total of 10,722 summer steelhead were captured at the Cowlitz Salmon Hatchery in 2012, which was the largest return since 2008. River flows during much of the study period were similar to 2008–2011 average flows, however, high-flow periods in July and November 2012 were nearly twice as high as the 2008–2011 average flows. We determined that 50 percent (273 fish) of the recycled steelhead returned to the hatchery and 18 percent (102 fish) of the recycled steelhead were caught by anglers. Most (243 fish; 89 percent) of the recycled steelhead that returned to the hatchery were recollected during July–August. The average elapsed time from release to recapture at the hatchery was 9 days (d) and 72 percent (182 fish) of the fish returned to the hatchery within 14 d of release. These trends were similar for recycled steelhead that were caught by anglers. Most fish were caught during July–August and the median time from release to capture was 10 d. We determined that 65 percent (70 fish) of the angler-caught fish returned to the hatchery within 14 d of release. River flows appeared to affect both hatchery returns and angler catch. The daily number of recycled steelhead that were recollected at the hatchery were low during periods when river flows were decreasing and high during periods when river flows were increasing. Conversely, daily angler catch of recycled steelhead generally was low when flows were increasing and high when flows were decreasing.We determined that 32 percent of the recycled steelhead (174 fish) were not removed from the lower Cowlitz River, based on observations from hatchery returns and angler reports, but results from the radio-tagged fish were insightful for understanding what may have happened to these fish. By comparison, we determined that 24 percent of the radio-tagged fish were not known to have been removed from the river. We determined that 12 percent of these fish were actively moving in the lower Cowlitz River during October 2012–January 2013. None of the radio-tagged fish were detected in tributaries during the study period except for a single fish that spent approximately 7 d in the Toutle River during early September. During October 2012–January 2013, 10 percent of the radio-tags from recycled steelhead were detected near popular fishing areas, and 2 percent of the radio-tagged steelhead were never detected during the study period. We suspect that a large proportion of these fish may have been harvested and not reported, or died.Detection patterns of radio-tagged steelhead showed that most fish (82 percent) moved upstream from the release site and were detected at the Trout Hatchery and the Barrier Dam sites. The median time from release to detection at these sites was 3.7 d and many of these fish made multiple trips between the two sites. Nearly one-third (29 percent) of the recycled steelhead that were detected at the Trout Hatchery and the Barrier Dam made at least two trips between the sites and some fish made as many as six trips. Radio-tagged fish that remained in the lower Cowlitz River during the spawning period (December 2012–January 2013) were observed in the river reach between the mouth of Ostrander Creek (river mile 10) and the Trout Hatchery (river mile 44).During this study, we collected data on opercle punch regrowth rates to understand the temporal effectiveness of this marking technique. We took opercle measurements for a total of 190 fish during the study. Fresh opercle punches were measured for 63 fish at the time of marking, and the remaining 127 fish were measured when fish returned to the hatchery. We determined that opercle punches remained open for about 30 d. The holes appeared to regrow slowly in the first 20 d after marking, but regrowth accelerated during the 20–30 d post-marking period. After 30 d, all opercle punches that we observed had completely closed due to tissue regrowth.Our study showed that a large proportion (68 percent) of the recycled steelhead were removed from the lower Cowlitz River. These fish primarily entered the hatchery or were caught by anglers within 14 d of release, which suggests that they present minimal risk to wild fish in the system. However, the remaining fish (32 percent) could not be accounted for, which may complicate fisheries management decisions associated with recycling summer steelhead. Findings from the radiotelemetry study suggest that unreported harvest or mortality could explain a large proportion of those fish that were not reported as having been removed from the river. Furthermore, intensive monitoring of the key spawning tributaries failed to detect a single fish during the spawning period. These findings were supported by observations from weir traps operated by the Washington Department of Fish and Wildlife. Our findings indicate that additional research may be warranted to further examine the effects of recycling hatchery summer steelhead in the lower Cowlitz River.

Radar transponder apparatus and signal processing technique

DOEpatents

Axline, Jr., Robert M.; Sloan, George R.; Spalding, Richard E.

1996-01-01

An active, phase-coded, time-grating transponder and a synthetic-aperture radar (SAR) and signal processor means, in combination, allow the recognition and location of the transponder (tag) in the SAR image and allow communication of information messages from the transponder to the SAR. The SAR is an illuminating radar having special processing modifications in an image-formation processor to receive an echo from a remote transponder, after the transponder receives and retransmits the SAR illuminations, and to enhance the transponder's echo relative to surrounding ground clutter by recognizing special transponder modulations from phase-shifted from the transponder retransmissions. The remote radio-frequency tag also transmits information to the SAR through a single antenna that also serves to receive the SAR illuminations. Unique tag-modulation and SAR signal processing techniques, in combination, allow the detection and precise geographical location of the tag through the reduction of interfering signals from ground clutter, and allow communication of environmental and status information from said tag to be communicated to said SAR.

Radar transponder apparatus and signal processing technique

DOEpatents

Axline, R.M. Jr.; Sloan, G.R.; Spalding, R.E.

1996-01-23

An active, phase-coded, time-grating transponder and a synthetic-aperture radar (SAR) and signal processor means, in combination, allow the recognition and location of the transponder (tag) in the SAR image and allow communication of information messages from the transponder to the SAR. The SAR is an illuminating radar having special processing modifications in an image-formation processor to receive an echo from a remote transponder, after the transponder receives and retransmits the SAR illuminations, and to enhance the transponder`s echo relative to surrounding ground clutter by recognizing special transponder modulations from phase-shifted from the transponder retransmissions. The remote radio-frequency tag also transmits information to the SAR through a single antenna that also serves to receive the SAR illuminations. Unique tag-modulation and SAR signal processing techniques, in combination, allow the detection and precise geographical location of the tag through the reduction of interfering signals from ground clutter, and allow communication of environmental and status information from said tag to be communicated to said SAR. 4 figs.

Metal Stable Isotope Tagging: Renaissance of Radioimmunoassay for Multiplex and Absolute Quantification of Biomolecules.

PubMed

Liu, Rui; Zhang, Shixi; Wei, Chao; Xing, Zhi; Zhang, Sichun; Zhang, Xinrong

2016-05-17

The unambiguous quantification of biomolecules is of great significance in fundamental biological research as well as practical clinical diagnosis. Due to the lack of a detectable moiety, the direct and highly sensitive quantification of biomolecules is often a "mission impossible". Consequently, tagging strategies to introduce detectable moieties for labeling target biomolecules were invented, which had a long and significant impact on studies of biomolecules in the past decades. For instance, immunoassays have been developed with radioisotope tagging by Yalow and Berson in the late 1950s. The later languishment of this technology can be almost exclusively ascribed to the use of radioactive isotopes, which led to the development of nonradioactive tagging strategy-based assays such as enzyme-linked immunosorbent assay, fluorescent immunoassay, and chemiluminescent and electrochemiluminescent immunoassay. Despite great success, these strategies suffered from drawbacks such as limited spectral window capacity for multiplex detection and inability to provide absolute quantification of biomolecules. After recalling the sequences of tagging strategies, an apparent question is why not use stable isotopes from the start? A reasonable explanation is the lack of reliable means for accurate and precise quantification of stable isotopes at that time. The situation has changed greatly at present, since several atomic mass spectrometric measures for metal stable isotopes have been developed. Among the newly developed techniques, inductively coupled plasma mass spectrometry is an ideal technique to determine metal stable isotope-tagged biomolecules, for its high sensitivity, wide dynamic linear range, and more importantly multiplex and absolute quantification ability. Since the first published report by our group, metal stable isotope tagging has become a revolutionary technique and gained great success in biomolecule quantification. An exciting research highlight in this area is the development and application of the mass cytometer, which fully exploited the multiplexing potential of metal stable isotope tagging. It realized the simultaneous detection of dozens of parameters in single cells, accurate immunophenotyping in cell populations, through modeling of intracellular signaling network and undoubted discrimination of function and connection of cell subsets. Metal stable isotope tagging has great potential applications in hematopoiesis, immunology, stem cells, cancer, and drug screening related research and opened a post-fluorescence era of cytometry. Herein, we review the development of biomolecule quantification using metal stable isotope tagging. Particularly, the power of multiplex and absolute quantification is demonstrated. We address the advantages, applicable situations, and limitations of metal stable isotope tagging strategies and propose suggestions for future developments. The transfer of enzymatic or fluorescent tagging to metal stable isotope tagging may occur in many aspects of biological and clinical practices in the near future, just as the revolution from radioactive isotope tagging to fluorescent tagging happened in the past.

Social Image Tag Ranking by Two-View Learning

NASA Astrophysics Data System (ADS)

Zhuang, Jinfeng; Hoi, Steven C. H.

Tags play a central role in text-based social image retrieval and browsing. However, the tags annotated by web users could be noisy, irrelevant, and often incomplete for describing the image contents, which may severely deteriorate the performance of text-based image retrieval models. In order to solve this problem, researchers have proposed techniques to rank the annotated tags of a social image according to their relevance to the visual content of the image. In this paper, we aim to overcome the challenge of social image tag ranking for a corpus of social images with rich user-generated tags by proposing a novel two-view learning approach. It can effectively exploit both textual and visual contents of social images to discover the complicated relationship between tags and images. Unlike the conventional learning approaches that usually assumes some parametric models, our method is completely data-driven and makes no assumption about the underlying models, making the proposed solution practically more effective. We formulate our method as an optimization task and present an efficient algorithm to solve it. To evaluate the efficacy of our method, we conducted an extensive set of experiments by applying our technique to both text-based social image retrieval and automatic image annotation tasks. Our empirical results showed that the proposed method can be more effective than the conventional approaches.

Security Techniques for Prevention of Rank Manipulation in Social Tagging Services including Robotic Domains

PubMed Central

2014-01-01

With smartphone distribution becoming common and robotic applications on the rise, social tagging services for various applications including robotic domains have advanced significantly. Though social tagging plays an important role when users are finding the exact information through web search, reliability and semantic relation between web contents and tags are not considered. Spams are making ill use of this aspect and put irrelevant tags deliberately on contents and induce users to advertise contents when they click items of search results. Therefore, this study proposes a detection method for tag-ranking manipulation to solve the problem of the existing methods which cannot guarantee the reliability of tagging. Similarity is measured for ranking the grade of registered tag on the contents, and weighted values of each tag are measured by means of synonym relevance, frequency, and semantic distances between tags. Lastly, experimental evaluation results are provided and its efficiency and accuracy are verified through them. PMID:25114975

Antibody enhancement of free-flow electrophoresis

NASA Technical Reports Server (NTRS)

Cohly, H. H. P.; Morrison, Dennis R.; Atassi, M. Zouhair

1988-01-01

Specific T cell clones and antibodies (ABs) were developed to study the efficiency of purifying closely associated T cells using Continuous Flow Electrophoresis System. Enhanced separation is accomplished by tagging cells first with ABs directed against the antigenic determinants on the cell surface and then with ABs against the Fc portion of the first AB. This second AB protrudes sufficiently beyond the cell membrane and glycocalyx to become the major overall cell surface potential determinant and thus causes a reduction of electrophoretic mobility. This project was divided into three phases. Phase one included development of specific T cell clones and separation of these specific clones. Phase two extends these principles to the separation of T cells from spleen cells and immunized lymph node cells. Phase three applies this double antibody technique to the separation of T cytotoxic cells from bone marrow.

Range-Depth Tracking of Sounds from a Single-Point Deployment by Exploiting the Deep-Water Sound Speed Minimum

DTIC Science & Technology

2015-09-30

dispersion of received signals with measured range. Two broad classes of calls are to be examined: deep-diving odontocetes such as sperm and potentially...comparison with satellite-tag positions (for sperm whales) or by comparison with ranges obtained via large-aperture vertical array techniques (for...depredating sperm whales, three of which had been tagged by satellite tags just before the deployment. Location fixes from the satellite tags are used to

Monitoring the Migrations of Wild Snake River Spring/Summer Chinook Salmon Juveniles, 2007-2008 Report of Research.

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

Achord, Stephen; Sandford, Benjamin P.; Hockersmith, Eric E.

2009-05-26

This report provides results from an ongoing project to monitor the migration behavior and survival of wild juvenile spring/summer Chinook salmon in the Snake River Basin. Data reported is from detections of PIT tagged fish during late summer 2007 through mid-2008. Fish were tagged in summer 2007 by the National Marine Fisheries Service (NMFS) in Idaho and by the Oregon Department of Fish and Wildlife (ODFW) in Oregon. Our analyses include migration behavior and estimated survival of fish at instream PIT-tag monitors and arrival timing and estimated survival to Lower Granite Dam. Principal results from tagging and interrogation during 2007-2008more » are listed below: (1) In July and August 2007, we PIT tagged and released 7,390 wild Chinook salmon parr in 12 Idaho streams or sample areas. (2) Overall observed mortality from collection, handling, tagging, and after a 24-hour holding period was 1.4%. (3) Of the 2,524 Chinook salmon parr PIT tagged and released in Valley Creek in summer 2007, 218 (8.6%) were detected at two instream PIT-tag monitoring systems in lower Valley Creek from late summer 2007 to the following spring 2008. Of these, 71.6% were detected in late summer/fall, 11.9% in winter, and 16.5% in spring. Estimated parr-to-smolt survival to Lower Granite Dam was 15.5% for the late summer/fall group, 48.0% for the winter group, and 58.5% for the spring group. Based on detections at downstream dams, the overall efficiency of VC1 (upper) or VC2 (lower) Valley Creek monitors for detecting these fish was 21.1%. Using this VC1 or VC2 efficiency, an estimated 40.8% of all summer-tagged parr survived to move out of Valley Creek, and their estimated survival from that point to Lower Granite Dam was 26.5%. Overall estimated parr-to-smolt survival for all summer-tagged parr from this stream at the dam was 12.1%. Development and improvement of instream PIT-tag monitoring systems continued throughout 2007 and 2008. (4) Testing of PIT-tag antennas in lower Big Creek during 2007-2008 showed these antennas (and anchoring method) are not adequate to withstand high spring flows in this drainage. Future plans involve removing these antennas before high spring flows. (5) At Little Goose Dam in 2008, length and/or weight were taken on 505 recaptured fish from 12 Idaho stream populations. Fish had grown an average of 40.1 mm in length and 10.6 g in weight over an average of 288 d. Their mean condition factor declined from 1.25 at release (parr) to 1.05 at recapture (smolt). (6) Mean release lengths for detected fish were significantly larger than for fish not detected the following spring and summer (P < 0.0001). (7) Fish that migrated through Lower Granite Dam in April and May were significantly larger at release than fish that migrated after May (P < 0.0001) (only 12 fish migrated after May). (8) In 2008, peak detections at Lower Granite Dam of parr tagged during summer 2007 (from the 12 stream populations in Idaho and 4 streams in Oregon) occurred during moderate flows of 87.5 kcfs on 7 May and high flows of 197.3 kcfs on 20 May. The 10th, 50th, and 90th percentile passage occurred on 30 April, 11 May, and 23 May, respectively. (9) In 2007-2008, estimated parr-to-smolt survival to Lower Granite Dam for Idaho and Oregon streams (combined) averaged 19.4% (range 6.2-38.4% depending on stream of origin). In Idaho streams the estimated parr-to-smolt survival averaged 21.0%. This survival was the second highest since 1993 for Idaho streams. Relative parr densities were lower in 2007 (2.4 parr/100 m{sup 2}) than in all previous years since 2000. In 2008, we observed low-to-moderate flows prior to mid-May and relatively cold weather conditions throughout the spring migration season. These conditions moved half of the fish through Lower Granite Dam prior to mid-May; then high flows moved 50 to 90% of the fish through the dam in only 12 days. Clearly, complex interrelationships of several factors drive the annual migrational timing of the stocks.« less

Cardiac MRI in mice at 9.4 Tesla with a transmit-receive surface coil and a cardiac-tailored intensity-correction algorithm.

PubMed

Sosnovik, David E; Dai, Guangping; Nahrendorf, Matthias; Rosen, Bruce R; Seethamraju, Ravi

2007-08-01

To evaluate the use of a transmit-receive surface (TRS) coil and a cardiac-tailored intensity-correction algorithm for cardiac MRI in mice at 9.4 Tesla (9.4T). Fast low-angle shot (FLASH) cines, with and without delays alternating with nutations for tailored excitation (DANTE) tagging, were acquired in 13 mice. An intensity-correction algorithm was developed to compensate for the sensitivity profile of the surface coil, and was tailored to account for the unique distribution of noise and flow artifacts in cardiac MR images. Image quality was extremely high and allowed fine structures such as trabeculations, valve cusps, and coronary arteries to be clearly visualized. The tag lines created with the surface coil were also sharp and clearly visible. Application of the intensity-correction algorithm improved signal intensity, tissue contrast, and image quality even further. Importantly, the cardiac-tailored properties of the correction algorithm prevented noise and flow artifacts from being significantly amplified. The feasibility and value of cardiac MRI in mice with a TRS coil has been demonstrated. In addition, a cardiac-tailored intensity-correction algorithm has been developed and shown to improve image quality even further. The use of these techniques could produce significant potential benefits over a broad range of scanners, coil configurations, and field strengths. (c) 2007 Wiley-Liss, Inc.

Individually Identifiable Surface Acoustic Wave Sensors, Tags and Systems

NASA Technical Reports Server (NTRS)

Hines, Jacqueline H. (Inventor); Solie, Leland P. (Inventor); Tucker, Dana Y. G. (Inventor); Hines, Andrew T. (Inventor)

2017-01-01

A surface-launched acoustic wave sensor tag system for remotely sensing and/or providing identification information using sets of surface acoustic wave (SAW) sensor tag devices is characterized by acoustic wave device embodiments that include coding and other diversity techniques to produce groups of sensors that interact minimally, reducing or alleviating code collision problems typical of prior art coded SAW sensors and tags, and specific device embodiments of said coded SAW sensor tags and systems. These sensor/tag devices operate in a system which consists of one or more uniquely identifiable sensor/tag devices and a wireless interrogator. The sensor device incorporates an antenna for receiving incident RF energy and re-radiating the tag identification information and the sensor measured parameter(s). Since there is no power source in or connected to the sensor, it is a passive sensor. The device is wirelessly interrogated by the interrogator.

Huntington II Simulation Program - TAG. Student Workbook, Teacher's Guide, and Resource Handbook.

ERIC Educational Resources Information Center

Friedland, James

Presented are instructions for the use of "TAG," a model for estimating animal population in a given area. The computer program asks the student to estimate the number of bass in a simulated farm pond using the technique of tagging and recovery. The objective of the simulation is to teach principles for estimating animal populations when they…

Single molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes

PubMed Central

Barlag, Britta; Beutel, Oliver; Janning, Dennis; Czarniak, Frederik; Richter, Christian P.; Kommnick, Carina; Göser, Vera; Kurre, Rainer; Fabiani, Florian; Erhardt, Marc; Piehler, Jacob; Hensel, Michael

2016-01-01

The investigation of the subcellular localization, dynamics and interaction of proteins and protein complexes in prokaryotes is complicated by the small size of the cells. Super-resolution microscopy (SRM) comprise various new techniques that allow light microscopy with a resolution that can be up to ten-fold higher than conventional light microscopy. Application of SRM techniques to living prokaryotes demands the introduction of suitable fluorescent probes, usually by fusion of proteins of interest to fluorescent proteins with properties compatible to SRM. Here we describe an approach that is based on the genetically encoded self-labelling enzymes HaloTag and SNAP-tag. Proteins of interest are fused to HaloTag or SNAP-tag and cell permeable substrates can be labelled with various SRM-compatible fluorochromes. Fusions of the enzyme tags to subunits of a type I secretion system (T1SS), a T3SS, the flagellar rotor and a transcription factor were generated and analysed in living Salmonella enterica. The new approach is versatile in tagging proteins of interest in bacterial cells and allows to determine the number, relative subcellular localization and dynamics of protein complexes in living cells. PMID:27534893

Diffusion in shear flow

NASA Astrophysics Data System (ADS)

Dufty, J. W.

1984-09-01

Diffusion of a tagged particle in a fluid with uniform shear flow is described. The continuity equation for the probability density describing the position of the tagged particle is considered. The diffusion tensor is identified by expanding the irreversible part of the probability current to first order in the gradient of the probability density, but with no restriction on the shear rate. The tensor is expressed as the time integral of a nonequilibrium autocorrelation function for the velocity of the tagged particle in its local fluid rest frame, generalizing the Green-Kubo expression to the nonequilibrium state. The tensor is evaluated from results obtained previously for the velocity autocorrelation function that are exact for Maxwell molecules in the Boltzmann limit. The effects of viscous heating are included and the dependence on frequency and shear rate is displayed explicitly. The mode-coupling contributions to the frequency and shear-rate dependent diffusion tensor are calculated.

Dual-Valve and Counter-Flow Surface Plasmon Resonance.

PubMed

Wang, Xiaoying; Zhou, Feimeng

2018-04-17

Two six-port injector valves and one selector valve commonly used in flow injection analysis are combined with a surface plasmon resonance (SPR) instrument wherein solutions introduced from the two inlets counter-flow inside the flow cell. The system is versatile as the same or different solutions can be rapidly and repeatedly introduced to the two fluidic channels in series or in parallel. Unlike most commercial SPR instruments employing a single injector valve, solutions separately injected from the two injector valves can be readily exchanged (<1 s) between the two channels. This new method, referred to as the alternate injection mode, not only saves analysis time but also facilitates efficient and facile surface reactions for ligand immobilization and prevents immobilized species from desorbing. These advantages are demonstrated with the measurements of binding of acetazolamide (222.2 Da) to histidine-tagged human carbonic anhydrase II (his-tagged HCA). Amine-containing residues of his-tagged HCA molecules tethered at Ni-nitrilotriacetic acid (NTA) sensors were rapidly cross-linked to the underlying carboxymethylated dextran. The higher ligand densities and more stable surfaces are essential for SPR detection of small molecule binding. In a different application, microglobulin solutions of increasing concentrations were introduced for continuous binding to the preimmobilized antibody. The kinetic and affinity measurements can be conducted without performing repeated dissociation and surface regeneration reactions.

Extending birthday paradox theory to estimate the number of tags in RFID systems.

PubMed

Shakiba, Masoud; Singh, Mandeep Jit; Sundararajan, Elankovan; Zavvari, Azam; Islam, Mohammad Tariqul

2014-01-01

The main objective of Radio Frequency Identification systems is to provide fast identification for tagged objects. However, there is always a chance of collision, when tags transmit their data to the reader simultaneously. Collision is a time-consuming event that reduces the performance of RFID systems. Consequently, several anti-collision algorithms have been proposed in the literature. Dynamic Framed Slotted ALOHA (DFSA) is one of the most popular of these algorithms. DFSA dynamically modifies the frame size based on the number of tags. Since the real number of tags is unknown, it needs to be estimated. Therefore, an accurate tag estimation method has an important role in increasing the efficiency and overall performance of the tag identification process. In this paper, we propose a novel estimation technique for DFSA anti-collision algorithms that applies birthday paradox theory to estimate the number of tags accurately. The analytical discussion and simulation results prove that the proposed method increases the accuracy of tag estimation and, consequently, outperforms previous schemes.

Extending Birthday Paradox Theory to Estimate the Number of Tags in RFID Systems

PubMed Central

Shakiba, Masoud; Singh, Mandeep Jit; Sundararajan, Elankovan; Zavvari, Azam; Islam, Mohammad Tariqul

2014-01-01

The main objective of Radio Frequency Identification systems is to provide fast identification for tagged objects. However, there is always a chance of collision, when tags transmit their data to the reader simultaneously. Collision is a time-consuming event that reduces the performance of RFID systems. Consequently, several anti-collision algorithms have been proposed in the literature. Dynamic Framed Slotted ALOHA (DFSA) is one of the most popular of these algorithms. DFSA dynamically modifies the frame size based on the number of tags. Since the real number of tags is unknown, it needs to be estimated. Therefore, an accurate tag estimation method has an important role in increasing the efficiency and overall performance of the tag identification process. In this paper, we propose a novel estimation technique for DFSA anti-collision algorithms that applies birthday paradox theory to estimate the number of tags accurately. The analytical discussion and simulation results prove that the proposed method increases the accuracy of tag estimation and, consequently, outperforms previous schemes. PMID:24752285

Bulk and molecular-level characterization of laboratory-aged biomass burning organic aerosol from oak leaf and heartwood fuels

NASA Astrophysics Data System (ADS)

Fortenberry, Claire F.; Walker, Michael J.; Zhang, Yaping; Mitroo, Dhruv; Brune, William H.; Williams, Brent J.

2018-02-01

The chemical complexity of biomass burning organic aerosol (BBOA) greatly increases with photochemical aging in the atmosphere, necessitating controlled laboratory studies to inform field observations. In these experiments, BBOA from American white oak (Quercus alba) leaf and heartwood samples was generated in a custom-built emissions and combustion chamber and photochemically aged in a potential aerosol mass (PAM) flow reactor. A thermal desorption aerosol gas chromatograph (TAG) was used in parallel with a high-resolution time-of-flight aerosol mass spectrometer (AMS) to analyze BBOA chemical composition at different levels of photochemical aging. Individual compounds were identified and integrated to obtain relative decay rates for key molecules. A recently developed chromatogram binning positive matrix factorization (PMF) technique was used to obtain mass spectral profiles for factors in TAG BBOA chromatograms, improving analysis efficiency and providing a more complete determination of unresolved complex mixture (UCM) components. Additionally, the recently characterized TAG decomposition window was used to track molecular fragments created by the decomposition of thermally labile BBOA during sample desorption. We demonstrate that although most primary (freshly emitted) BBOA compounds deplete with photochemical aging, certain components eluting within the TAG thermal decomposition window are instead enhanced. Specifically, the increasing trend in the decomposition m/z 44 signal (CO2+) indicates formation of secondary organic aerosol (SOA) in the PAM reactor. Sources of m/z 60 (C2H4O2+), typically attributed to freshly emitted BBOA in AMS field measurements, were also investigated. From the TAG chemical speciation and decomposition window data, we observed a decrease in m/z 60 with photochemical aging due to the decay of anhydrosugars (including levoglucosan) and other compounds, as well as an increase in m/z 60 due to the formation of thermally labile organic acids within the PAM reactor, which decompose during TAG sample desorption. When aging both types of BBOA (leaf and heartwood), the AMS data exhibit a combination of these two contributing effects, causing limited change to the overall m/z 60 signal. Our observations demonstrate the importance of chemically speciated data in fully understanding bulk aerosol measurements provided by the AMS in both laboratory and field studies.

Enhancing Cardiac Triacylglycerol Metabolism Improves Recovery From Ischemic Stress

PubMed Central

Liu, Li; Goldberg, Ira J.

2015-01-01

Elevated cardiac triacylglycerol (TAG) content is traditionally equated with cardiolipotoxicity and suggested to be a culprit in cardiac dysfunction. However, previous work demonstrated that myosin heavy-chain–mediated cardiac-specific overexpression of diacylglycerol transferase 1 (MHC-DGAT1), the primary enzyme for TAG synthesis, preserved cardiac function in two lipotoxic mouse models despite maintaining high TAG content. Therefore, we examined whether increased cardiomyocyte TAG levels due to DGAT1 overexpression led to changes in cardiac TAG turnover rates under normoxia and ischemia-reperfusion conditions. MHC-DGAT1 mice had elevated TAG content and synthesis rates, which did not alter cardiac function, substrate oxidation, or myocardial energetics. MHC-DGAT1 hearts had ischemia-induced lipolysis; however, when a physiologic mixture of long-chain fatty acids was provided, enhanced TAG turnover rates were associated with improved functional recovery from low-flow ischemia. Conversely, exogenous supply of palmitate during reperfusion suppressed elevated TAG turnover rates and impaired recovery from ischemia in MHC-DGAT1 hearts. Collectively, this study shows that elevated TAG content, accompanied by enhanced turnover, does not adversely affect cardiac function and, in fact, provides cardioprotection from ischemic stress. In addition, the results highlight the importance of exogenous supply of fatty acids when assessing cardiac lipid metabolism and its relationship with cardiac function. PMID:25858561

Advancing the surgical implantation of electronic tags in fish: a gap analysis and research agenda based on a review of trends in intracoelomic tagging effects studies

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

Cooke, Steven J.; Woodley, Christa M.; Eppard, M. B.

2011-03-08

Early approaches to surgical implantation of electronic tags in fish were often through trial and error, however, in recent years there has been an interest in using scientific research to identify techniques and procedures that improve the outcome of surgical procedures and determine the effects of tagging on individuals. Here we summarize the trends in 108 peer-reviewed electronic tagging effect studies focused on intracoleomic implantation to determine opportunities for future research. To date, almost all of the studies have been conducted in freshwater, typically in laboratory environments, and have focused on biotelemetry devices. The majority of studies have focused onmore » salmonids, cyprinids, ictalurids and centrarchids, with a regional bias towards North America, Europe and Australia. Most studies have focused on determining whether there is a negative effect of tagging relative to control fish, with proportionally fewer that have contrasted different aspects of the surgical procedure (e.g., methods of sterilization, incision location, wound closure material) that could advance the discipline. Many of these studies included routine endpoints such as mortality, growth, healing and tag retention, with fewer addressing sublethal measures such as swimming ability, predator avoidance, physiological costs, or fitness. Continued research is needed to further elevate the practice of electronic tag implantation in fish in order to ensure that the data generated are relevant to untagged conspecifics (i.e., no long-term behavioural or physiological consequences) and the surgical procedure does not impair the health and welfare status of the tagged fish. To that end, we advocate for i) rigorous controlled manipulations based on statistical designs that have adequate power, account for inter-individual variation, and include controls and shams, ii) studies that transcend the laboratory and the field with more studies in marine waters, iii) incorporation of knowledge and techniques emerging from the medical and veterinary disciplines, iv) addressing all components of the surgical event, v) comparative studies that evaluate the same surgical techniques on multiple species and in different environments, vi) consideration of how biotic factors (e.g., sex, age, size) influence tagging outcomes, and vii) studies that cover a range of endpoints over ecologically-relevant time periods.« less

Tracing the contribution of debris flow-dominated channels to gravel-bed torrential river channel: implementing pit-tags in the upper Guil River (French Alps)

NASA Astrophysics Data System (ADS)

Arnaud-Fassetta, Gilles; Lissak, Candide; Fort, Monique; Bétard, François; Carlier, Benoit; Cossart, Etienne; Madelin, Malika; Viel, Vincent; Charnay, Bérengère; Bletterie, Xavier

2014-05-01

In the upper, wider reaches of Alpine valleys, shaping of active channels is usually subject to rapid change. It mostly depends upon hydro-climatic variability, runoff concentration and sediment supply, and may result in alternating sequences of fluvial and debris-flow pulses, as recorded in alluvial fans and terraces. Our study, carried in the frame of SAMCO (ANR) project, focuses on the upper Guil River Valley (Queyras, Southern French Alps) cut into the slaty shale "schistes lustrés". Steep, lower order drains carry a contrasted solid discharge, including predominantly sandy-loam particles mixed with gravels and boulders (sandstone schists, ophiolites). Abundant sediment supply by frost shattering, snow avalanche and landslides is then reworked during snowmelt or summer storm runoff events, and may result in catastrophic, very destructive floods along the main channel, as shown by historical records. Following the RI-30 year 2000 flood, our investigations included sediment budgets, i.e. balance of erosion and deposition, and the mapping of the source, transport and storage of various sediments (talus, colluvium, torrential fans, terraces). To better assess sediment fluxes and sediment delivery into the main channel network, we implemented tracers (pit-tags) in selected sub-catchments, significantly contributing to the sediment yield of the valley bottoms during the floods and/or avalanches: Maloqueste, Combe Morel, Bouchouse and Peyronnelle catchments. The first three are direct tributaries of the Guil River whereas the Peyronnelle is a left bank tributary of the Peynin River, which joins the Guil River via an alluvial cone with high human and material stakes. The Maloqueste and the Combe Morel are two tributaries facing each other in the Guil valley, representing a double lateral constraint for the road during flood events of the Guil River. After pit-tag initialisation in laboratory, we set them up along the four tributaries: Maloqueste (20 pit-tags), Combe Morel (20 pit-tags), Bouchouse (40 pit-tags), Peyronnelle (100 pit-tags). All pit-tags were installed high upstream enough in each catchment to avoid their loss and/or burying, and permit their monitoring over several years. The grain size (D50 = 30-220 cm) of the selected blocks reflects the average size of the bed load. Smaller blocks were discarded to avoid losing their track from the first event. The blocks were positioned according to two logics: (i) Longitudinally, we implemented every 20-30 m a series of nested blocks into the channel; (ii) Transversally, we selected blocks placed in low and high position to quantify the transport dynamics during events of low (torrential) and high (debris flow) magnitude. The sites were mapped at high resolution, and blocks were spotted thanks to fluorescent painting, GPS surveys and detailed photographs. We emplaced the pit-tags in September 2013, before expected avalanches and debris flows during winter and spring 2014. Pit-tags detection, Lidar and photogrammetric surveys are planned for early summer 2014, during which we expect quantifying movement, distance and sorting of blocks along the thalwegs during "ordinary" snowmelt runoff. We intend to continue this monitoring long enough to put observed current and future dynamics in relation to short-term climatic changes.

Velocity measurement in rocket exhaust and general aerodynamic flows by photolysis of H2O and laser induced fluorescence of OH

NASA Technical Reports Server (NTRS)

Boedeker, Laurence R.

1992-01-01

A 'tagging' approach in which the photolysis of H2O by an excimer laser creates a zone of enhanced OH concentration, while a second, pulsed-UV laser detects tagged-zone convection via time-delayed excitation of OH fluorescence, depends on the photodissociation process and the kinetics of OH decay (relative to velocity). For application to the fuel-rich, high supersonic Mach number exhaust flow of the SSME, the detection of OH is being accomplished with either a pulsed narrowband UV dye laser or a tunable XeCl excimer laser for excitation of an OH 0-0 band transition, while the two-photon photolysis of H2O is conducted by focusing an injection-locked KrF excimer laser into the flow.

Dual functionalized graphene oxide serves as a carrier for delivering oligohistidine- and biotin-tagged biomolecules into cells.

PubMed

Jana, Batakrishna; Mondal, Goutam; Biswas, Atanu; Chakraborty, Indrani; Saha, Abhijit; Kurkute, Prashant; Ghosh, Surajit

2013-11-01

A versatile method of dual chemical functionalization of graphene oxide (GO) with Tris-[nitrilotris(acetic acid)] (Tris-NTA) and biotin for cellular delivery of oligohistidine- and biotin-tagged biomolecules is reported. Orthogonally functionalized GO surfaces with Tris-NTA and biotin to obtain a dual-functionalized GO (DFGO) are prepared and characterized by various spectroscopic and microscopic techniques. Fluorescence microscopic images reveal that DFGO surfaces are capable of binding oligohistidine-tagged biomolecules/proteins and avidin/biotin-tagged biomolecules/proteins orthogonally. The DFGO nanoparticles are non-cytotoxic in nature and can deliver oligohistidine- and biotin-tagged biomolecules simultaneously into the cell. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Method of remote powering and detecting multiple UWB passive tags in an RFID system

DOEpatents

Dowla, Farid U [Castro Valley, CA; Nekoogar, Faranak [San Ramon, CA; Benzel, David M [Livermore, CA; Dallum, Gregory E [Livermore, CA; Spiridon, Alex [Palo Alto, CA

2012-05-29

A new Radio Frequency Identification (RFID), tracking, powering apparatus/system and method using coded Ultra-wideband (UWB) signaling is introduced. The proposed hardware and techniques disclosed herein utilize a plurality of passive UWB transponders in a field of an RFID-radar system. The radar system itself enables multiple passive tags to be remotely powered (activated) at about the same time frame via predetermined frequency UWB pulsed formats. Once such tags are in an activated state, an UWB radar transmits specific "interrogating codes" to put predetermined tags in an awakened status. Such predetermined tags can then communicate by a unique "response code" so as to be detected by an UWB system using radar methods.

Parasites as biological tags to assess host population structure: Guidelines, recent genetic advances and comments on a holistic approach☆

PubMed Central

Catalano, Sarah R.; Whittington, Ian D.; Donnellan, Stephen C.; Gillanders, Bronwyn M.

2013-01-01

We review the use of parasites as biological tags of marine fishes and cephalopods in host population structure studies. The majority of the work published has focused on marine fish and either single parasite species or more recently, whole parasite assemblages, as biological tags. There is representation of host organisms and parasites from a diverse range of taxonomic groups, although focus has primarily been on host species of commercial importance. In contrast, few studies have used parasites as tags to assess cephalopod population structure, even though records of parasites infecting cephalopods are well-documented. Squid species are the only cephalopod hosts for which parasites as biological tags have been applied, with anisakid nematode larvae and metacestodes being the parasite taxa most frequently used. Following a brief insight into the importance of accurate parasite identification, the population studies that have used parasites as biological tags for marine fishes and cephalopods are reviewed, including comments on the dicyemid mesozoans. The advancement of molecular genetic techniques is discussed in regards to the new ways parasite genetic data can be incorporated into population structure studies, alongside host population genetic analyses, followed by an update on the guidelines for selecting a parasite species as a reliable tag candidate. As multiple techniques and methods can be used to assess the population structure of marine organisms (e.g. artificial tags, phenotypic characters, biometrics, life history, genetics, otolith microchemistry and parasitological data), we conclude by commenting on a holistic approach to allow for a deeper insight into population structuring. PMID:25197624

Antibiotic use during the intracoelomic implantation of electronic tags into fish

USGS Publications Warehouse

Mulcahy, D.M.

2011-01-01

The use of antibiotics, in particular, the use of a single dose of antibiotics during electronic tag implantation is of unproven value, and carries with it the potential for the development of antibiotic resistance in bacteria and the alteration of the immune response of the fish. Antibiotic use during electronic tag implantation must conform to relevant drug laws and regulations in the country where work is being done, including the requirements for withdrawal times before human consumption is a possibility. Currently, the choice of antibiotics (most often tetracycline or oxytetracycline) and the use of a single dose of the drug are decisions made without knowledge of the basic need for antibiotic usage and of the bacteria involved in infections that occur following electronic tag implantation. Correct perioperative use of an antibiotic is to apply the drug to the animal before surgery begins, to assure serum and tissue levels of the drug are adequate before the incision is made. However, the most common perioperative application of antibiotics during implantation of an electronic tag is to delay the administration of the drug, injecting it into the coelom after the electronic tag is inserted, just prior to closure of the incision. There is little empirical evidence that the present application of antibiotics in fish being implanted with electronic tags is of value. Improvements should first be made to surgical techniques, especially the use of aseptic techniques and sterilized instruments and electronic tags, before resorting to antibiotics. ?? 2010 Springer Science+Business Media B.V.(outside the USA).

Seal Whiskers Vibrate Over Broad Frequencies During Hydrodynamic Tracking.

PubMed

Murphy, Christin T; Reichmuth, Colleen; Eberhardt, William C; Calhoun, Benton H; Mann, David A

2017-08-21

Although it is known that seals can use their whiskers (vibrissae) to extract relevant information from complex underwater flow fields, the underlying functioning of the system and the signals received by the sensors are poorly understood. Here we show that the vibrations of seal whiskers may provide information about hydrodynamic events and enable the sophisticated wake-tracking abilities of these animals. We developed a miniature accelerometer tag to study seal whisker movement in situ. We tested the ability of the tag to measure vibration in excised whiskers in a flume in response to laminar flow and disturbed flow. We then trained a seal to wear the tag and follow an underwater hydrodynamic trail to measure the whisker signals available to the seal. The results showed that whiskers vibrated at frequencies of 100-300 Hz, with a dynamic response. These measurements are the first to capture the incoming signals received by the vibrissae of a live seal and show that there are prominent signals at frequencies where the seal tactogram shows good sensitivity. Tapping into the mechanoreceptive interface between the animal and the environment may help to decipher the functional basis of this extraordinary hydrodynamic detection ability.

Tracking animals in freshwater with electronic tags: past, present and future

USGS Publications Warehouse

Cooke, Steven J.; Midwood, Jonathan D.; Thiem, Jason D.; Klimley, Peter; Lucas, Martyn C.; Thorstad, Eva B.; Eiler, John; Holbrook, Chris; Ebner, Brendan C.

2013-01-01

Considerable technical developments over the past half century have enabled widespread application of electronic tags to the study of animals in the wild, including in freshwater environments. We review the constraints associated with freshwater telemetry and biologging and the technical developments relevant to their use. Technical constraints for tracking animals are often influenced by the characteristics of the animals being studied and the environment they inhabit. Collectively, they influence which and how technologies can be used and their relative effectiveness. Although radio telemetry has historically been the most commonly used technology in freshwater, passive integrated transponder (PIT) technology, acoustic telemetry and biologgers are becoming more popular. Most telemetry studies have focused on fish, although an increasing number have focused on other taxa, such as turtles, crustaceans and molluscs. Key technical developments for freshwater systems include: miniaturization of tags for tracking small-size life stages and species, fixed stations and coded tags for tracking large samples of animals over long distances and large temporal scales, inexpensive PIT systems that enable mass tagging to yield population- and community-level relevant sample sizes, incorporation of sensors into electronic tags, validation of tag attachment procedures with a focus on maintaining animal welfare, incorporation of different techniques (for example, genetics, stable isotopes) and peripheral technologies (for example, geographic information systems, hydroacoustics), development of novel analytical techniques, and extensive international collaboration. Innovations are still needed in tag miniaturization, data analysis and visualization, and in tracking animals over larger spatial scales (for example, pelagic areas of lakes) and in challenging environments (for example, large dynamic floodplain systems, under ice). There seems to be a particular need for adapting various global positioning system and satellite tagging approaches to freshwater. Electronic tagging provides a mechanism to collect detailed information from imperilled animals and species that have no direct economic value. Current and future advances will continue to improve our knowledge of the natural history of aquatic animals and ecological processes in freshwater ecosystems while facilitating evidence-based resource management and conservation.

Quantitative chemical tagging, stellar ages and the chemo-dynamical evolution of the Galactic disc

NASA Astrophysics Data System (ADS)

Mitschang, A. W.; De Silva, G.; Zucker, D. B.; Anguiano, B.; Bensby, T.; Feltzing, S.

2014-03-01

The early science results from the new generation of high-resolution stellar spectroscopic surveys, such as Galactic Archaeology with HERMES (GALAH) and the Gaia European Southern Observatory survey (Gaia-ESO), will represent major milestones in the quest to chemically tag the Galaxy. Yet this technique to reconstruct dispersed coeval stellar groups has remained largely untested until recently. We build on previous work that developed an empirical chemical tagging probability function, which describes the likelihood that two field stars are conatal, that is, they were formed in the same cluster environment. In this work, we perform the first ever blind chemical tagging experiment, i.e. tagging stars with no known or otherwise discernible associations, on a sample of 714 disc field stars with a number of high-quality high-resolution homogeneous metal abundance measurements. We present evidence that chemical tagging of field stars does identify coeval groups of stars, yet these groups may not represent distinct formation sites, e.g. as in dissolved open clusters, as previously thought. Our results point to several important conclusions, among them that group finding will be limited strictly to chemical abundance space, e.g. stellar ages, kinematics, colours, temperature and surface gravity do not enhance the detectability of groups. We also demonstrate that in addition to its role in probing the chemical enrichment and kinematic history of the Galactic disc, chemical tagging represents a powerful new stellar age determination technique.

Differentiation of tumor from viable myocardium using cardiac tagging with MR imaging.

PubMed

Bouton, S; Yang, A; McCrindle, B W; Kidd, L; McVeigh, E R; Zerhouni, E A

1991-01-01

We report the application of myocardial tagging by MR to define tissue planes and differentiate contractile from noncontractile tissue in a neonate with congenital cardiac rhabdomyoma. Using custom-written pulse programming software, six 2 mm thick radiofrequency (RF) slice-selective presaturation pulses (tags) were used to label the chest wall and myocardium in a star pattern in diastole, approximately 60 ms before the R-wave gating trigger. This method successfully delineated the myocardium from noncontractile tumor, providing information that influenced clinical management. This RF tagging technique allowed us to confirm the exact intramyocardial location of a congenital cardiac tumor.

Efficient radial tagging CMR exam: A coherent k-space reading and image reconstruction approach.

PubMed

Golshani, Shokoufeh; Nasiraei-Moghaddam, Abbas

2017-04-01

Cardiac MR tagging techniques, which facilitate the strain evaluation, have not yet been widely adopted in clinics due to inefficiencies in acquisition and postprocessing. This problem may be alleviated by exploiting the coherency in the three steps of tagging: preparation, acquisition, and reconstruction. Herein, we propose a fully polar-based tagging approach that may lead to real-time strain mapping. Radial readout trajectories were used to acquire radial tagging images and a Hankel-based algorithm, referred to as Polar Fourier Transform (PFT), has been adapted for reconstruction of the acquired raw data. In both phantom and human subjects, the overall performance of the method was investigated against radial undersampling and compared with the conventional reconstruction methods. Radially tagged images were reconstructed by the proposed PFT method from as few as 24 spokes with normalized root-mean-square-error of less than 3%. The reconstructed images showed a central focusing behavior, where the undersampling effects were pushed to the peripheral areas out of the central region of interest. Comparing the results with the re-gridding reconstruction technique, superior image quality and high robustness of the method were further established. In addition, a relative increase of 68 ± 2.5% in tagline sharpness was achieved for the PFT images and also higher tagging contrast (72 ± 5.6%), resulted from the well-tolerated undersampling artifacts, was observed in all reconstructions. The proposed approach led to the acceleration of the acquisition process, which was evaluated for up to eight-fold retrospectively from the fully sampled data. This is promising toward real-time imaging, and in contrast to iterative techniques, the method is consistent with online reconstruction. Magn Reson Med 77:1459-1472, 2017. © 2016 International Society for Magnetic Resonance in Medicine. © 2016 International Society for Magnetic Resonance in Medicine.

Correlative light and electron microscopic detection of GFP-labeled proteins using modular APEX.

PubMed

Ariotti, Nicholas; Hall, Thomas E; Parton, Robert G

2017-01-01

The use of green fluorescent protein (GFP) and related proteins has revolutionized light microscopy. Here we describe a rapid and simple method to localize GFP-tagged proteins in cells and in tissues by electron microscopy (EM) using a modular approach involving a small GFP-binding peptide (GBP) fused to the ascorbate peroxidase-derived APEX2 tag. We provide a method for visualizing GFP-tagged proteins by light and EM in cultured cells and in the zebrafish using modular APEX-GBP. Furthermore, we describe in detail the benefits of this technique over many of the currently available correlative light and electron microscopy approaches and demonstrate APEX-GBP is readily applicable to modern three-dimensional techniques. Copyright © 2017 Elsevier Inc. All rights reserved.

InkTag: Secure Applications on an Untrusted Operating System

PubMed Central

Hofmann, Owen S.; Kim, Sangman; Dunn, Alan M.; Lee, Michael Z.; Witchel, Emmett

2014-01-01

InkTag is a virtualization-based architecture that gives strong safety guarantees to high-assurance processes even in the presence of a malicious operating system. InkTag advances the state of the art in untrusted operating systems in both the design of its hypervisor and in the ability to run useful applications without trusting the operating system. We introduce paraverification, a technique that simplifies the InkTag hypervisor by forcing the untrusted operating system to participate in its own verification. Attribute-based access control allows trusted applications to create decentralized access control policies. InkTag is also the first system of its kind to ensure consistency between secure data and metadata, ensuring recoverability in the face of system crashes. PMID:24429939

InkTag: Secure Applications on an Untrusted Operating System.

PubMed

Hofmann, Owen S; Kim, Sangman; Dunn, Alan M; Lee, Michael Z; Witchel, Emmett

2013-01-01

InkTag is a virtualization-based architecture that gives strong safety guarantees to high-assurance processes even in the presence of a malicious operating system. InkTag advances the state of the art in untrusted operating systems in both the design of its hypervisor and in the ability to run useful applications without trusting the operating system. We introduce paraverification , a technique that simplifies the InkTag hypervisor by forcing the untrusted operating system to participate in its own verification. Attribute-based access control allows trusted applications to create decentralized access control policies. InkTag is also the first system of its kind to ensure consistency between secure data and metadata, ensuring recoverability in the face of system crashes.

Depth discrimination in acousto-optic cerebral blood flow measurement simulation

NASA Astrophysics Data System (ADS)

Tsalach, A.; Schiffer, Z.; Ratner, E.; Breskin, I.; Zeitak, R.; Shechter, R.; Balberg, M.

2016-03-01

Monitoring cerebral blood flow (CBF) is crucial, as inadequate perfusion, even for relatively short periods of time, may lead to brain damage or even death. Thus, significant research efforts are directed at developing reliable monitoring tools that will enable continuous, bed side, simple and cost-effective monitoring of CBF. All existing non invasive bed side monitoring methods, which are mostly NIRS based, such as Laser Doppler or DCS, tend to underestimate CBF in adults, due to the indefinite effect of extra-cerebral tissues on the obtained signal. If those are to find place in day to day clinical practice, the contribution of extra-cerebral tissues must be eliminated and data from the depth (brain) should be extracted and discriminated. Recently, a novel technique, based on ultrasound modulation of light was developed for non-invasive, continuous CBF monitoring (termed ultrasound-tagged light (UTL or UT-NIRS)), and shown to correlate with readings of 133Xe SPECT and laser Doppler. We have assembled a comprehensive computerized simulation, modeling this acousto-optic technique in a highly scattering media. Using the combination of light and ultrasound, we show how depth information may be extracted, thus distinguishing between flow patterns taking place at different depths. Our algorithm, based on the analysis of light modulated by ultrasound, is presented and examined in a computerized simulation. Distinct depth discrimination ability is presented, suggesting that using such method one can effectively nullify the extra-cerebral tissues influence on the obtained signals, and specifically extract cerebral flow data.

3D micro-crack propagation simulation at enamel/adhesive interface using FE submodeling and element death techniques.

PubMed

Liu, Heng-Liang; Lin, Chun-Li; Sun, Ming-Tsung; Chang, Yen-Hsiang

2010-06-01

This study investigates micro-crack propagation at the enamel/adhesive interface using finite element (FE) submodeling and element death techniques. A three-dimensional (3D) FE macro-model of the enamel/adhesive/ceramic subjected to shear bond testing was generated and analyzed. A 3D micro-model with interfacial bonding structure was constructed at the upper enamel/adhesive interface where the stress concentration was found from the macro-model results. The morphology of this interfacial bonding structure (i.e., resin tag) was assigned based on resin tag geometry and enamel rod arrangement from a scanning electron microscopy micrograph. The boundary conditions for the micro-model were determined from the macro-model results. A custom iterative code combined with the element death technique was used to calculate the micro-crack propagation. Parallel experiments were performed to validate this FE simulation. The stress concentration within the adhesive occurred mainly at the upper corner near the enamel/adhesive interface and the resin tag base. A simulated fracture path was found at the resin tag base along the enamel/adhesive interface. A morphological observation of the fracture patterns obtained from in vitro testing corresponded with the simulation results. This study shows that the FE submodeling and element death techniques could be used to simulate the 3D micro-stress pattern and the crack propagation noted at the enamel/adhesive interface.

Development of N_2O-MTV for low-speed flow and in-situ deployment to an integral effect test facility

NASA Astrophysics Data System (ADS)

André, Matthieu A.; Burns, Ross A.; Danehy, Paul M.; Cadell, Seth R.; Woods, Brian G.; Bardet, Philippe M.

2018-01-01

A molecular tagging velocity (MTV) technique is developed to non-intrusively measure velocity in an integral effect test (IET) facility simulating a high-temperature helium-cooled nuclear reactor in accident scenarios. In these scenarios, the velocities are expected to be low, on the order of 1 m/s or less, which forces special requirements on the MTV tracer selection. Nitrous oxide (N_2O) is identified as a suitable seed gas to generate NO tracers capable of probing the flow over a large range of pressure, temperature, and flow velocity. The performance of N_2O-MTV is assessed in the laboratory at temperature and pressure ranging from 295 to 781 K and 1 to 3 atm. MTV signal improves with a temperature increase, but decreases with a pressure increase. Velocity precision down to 0.004 m/s is achieved with a probe time of 40 ms at ambient pressure and temperature. Measurement precision is limited by tracer diffusion, and absorption of the tag laser beam by the seed gas. Processing by cross-correlation of single-shot images with high signal-to-noise ratio reference images improves the precision by about 10% compared to traditional single-shot image correlations. The instrument is then deployed to the IET facility. Challenges associated with heat, vibrations, safety, beam delivery, and imaging are addressed in order to successfully operate this sensitive instrument in-situ. Data are presented for an isothermal depressurized conduction cooldown. Velocity profiles from MTV reveal a complex flow transient driven by buoyancy, diffusion, and instability taking place over short (<1 s) and long (>30 min) time scales at sub-meter per second speed. The precision of the in-situ results is estimated at 0.027, 0.0095, and 0.006 m/s for a probe time of 5, 15, and 35 ms, respectively.

Use of passive scalar tagging for the study of coherent structures in the plane mixing layer

NASA Technical Reports Server (NTRS)

Ramaprian, B. R.; Sandham, N. D.; Mungal, M. G.; Reynolds, W. C.

1988-01-01

Data obtained from the numerical simulation of a 2-D mixing layer were used to study the feasibility of using the instantaneous concentration of a passive scalar for detecting the typical coherent structures in the flow. The study showed that this technique works quite satisfactorily and yields results similar to those that can be obtained by using the instantaneous vorticity for structure detection. Using the coherent events educed by the scalar conditioning technique, the contribution of the coherent events to the total turbulent momentum and scalar transport was estimated. It is found that the contribution from the typical coherent events is of the same order as that of the time-mean value. However, the individual contributions become very large during the pairing of these structures. The increase is particularly spectacular in the case of the Reynolds shear stress.

Alignment of leading-edge and peak-picking time of arrival methods to obtain accurate source locations

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

Roussel-Dupre, R.; Symbalisty, E.; Fox, C.

2009-08-01

The location of a radiating source can be determined by time-tagging the arrival of the radiated signal at a network of spatially distributed sensors. The accuracy of this approach depends strongly on the particular time-tagging algorithm employed at each of the sensors. If different techniques are used across the network, then the time tags must be referenced to a common fiducial for maximum location accuracy. In this report we derive the time corrections needed to temporally align leading-edge, time-tagging techniques with peak-picking algorithms. We focus on broadband radio frequency (RF) sources, an ionospheric propagation channel, and narrowband receivers, but themore » final results can be generalized to apply to any source, propagation environment, and sensor. Our analytic results are checked against numerical simulations for a number of representative cases and agree with the specific leading-edge algorithm studied independently by Kim and Eng (1995) and Pongratz (2005 and 2007).« less

Signal and array processing techniques for RFID readers

NASA Astrophysics Data System (ADS)

Wang, Jing; Amin, Moeness; Zhang, Yimin

2006-05-01

Radio Frequency Identification (RFID) has recently attracted much attention in both the technical and business communities. It has found wide applications in, for example, toll collection, supply-chain management, access control, localization tracking, real-time monitoring, and object identification. Situations may arise where the movement directions of the tagged RFID items through a portal is of interest and must be determined. Doppler estimation may prove complicated or impractical to perform by RFID readers. Several alternative approaches, including the use of an array of sensors with arbitrary geometry, can be applied. In this paper, we consider direction-of-arrival (DOA) estimation techniques for application to near-field narrowband RFID problems. Particularly, we examine the use of a pair of RFID antennas to track moving RFID tagged items through a portal. With two antennas, the near-field DOA estimation problem can be simplified to a far-field problem, yielding a simple way for identifying the direction of the tag movement, where only one parameter, the angle, needs to be considered. In this case, tracking of the moving direction of the tag simply amounts to computing the spatial cross-correlation between the data samples received at the two antennas. It is pointed out that the radiation patterns of the reader and tag antennas, particularly their phase characteristics, have a significant effect on the performance of DOA estimation. Indoor experiments are conducted in the Radar Imaging and RFID Labs at Villanova University for validating the proposed technique for target movement direction estimations.

Movements of yellow perch marked in southern Green Bay, Lake Michigan, in 1950

USGS Publications Warehouse

Mraz, Donald

1952-01-01

These investigations have demonstrated that for studying migration of yellow perch, tagging is superior to fin-clipping as a method of marking. The technique of the tagging, however, needs to be improved, and better means must be found to trace tagged fish of small size. Furthermore certain small regions in the bay not at present open to commercial fishing must be explored in order to obtain more comprehensive information as to migration.

Radio-tracer techniques for the study of flow in saturated porous materials

USGS Publications Warehouse

Skibitzke, H.E.; Chapman, H.T.; Robinson, G.M.; McCullough, Richard A.

1961-01-01

An experiment was conducted by the U.S. Geological Survey to determine the feasibility of using a radioactive substance as a tracer in the study of microscopic flow in a saturated porous solid. A radioactive tracer was chosen in preference to dye or other chemical in order to eliminate effects of the tracer itself on the flow system such as those relating to density, viscosity and surface tension. The porous solid was artificial "sandstone" composed of uniform fine grains of sand bonded together with an epoxy adhesive. The sides of the block thus made were sealed with an epoxy coating compound to insure water-tightness. Because of the chemical inertness of the block it was possible to use radioactive phosphorus (P32). Ion-exchange equilibrium was created between the block and nonradioactive phosphoric acid. Then a tracer tagged with P32 was injected into the block in the desired geometric configuration, in this case, a line source. After equilibrium in isotopic exchange was reached between the block and the line source, the block was rinsed, drained and sawn into slices. It was found that a quantitative analysis of the flow system may be made by assaying the dissected block. ?? 1961.

Turbulent channel flow under moderate polymer drag reduction

NASA Astrophysics Data System (ADS)

Elsnab, John; Monty, Jason; White, Christopher; Koochesfahani, Manoochehr; Klewicki, Joseph

2017-11-01

Streamwise velocity profiles and their wall-normal derivatives are used to investigate the properties of turbulent channel flow under the moderate polymer drag reduction (DR) conditions of 6-27%. Velocity data were obtained over a friction Reynolds number (Re) from 650-1800 using the single velocity component version of molecular tagging velocimetry (MTV). This adaptation of the MTV technique captures instantaneous profiles at high spatial resolution (>800 data points per profile), thus generating well-resolved derivative information. The mean velocity profiles indicate that the extent of the logarithmic region diminishes with increasing polymer concentration, while the logarithmic profile slope increases for drag reductions greater than about 20%. The measurements allow reconstruction of the mean momentum balance for channel flow that provides additional insights regarding the physics described by previous numerical simulation analyses that examined the mean dynamical structure of polymer laden channel flow at low Re. The present findings indicate that the polymer modifies the onset of the inertial domain, and that the extent of this domain shrinks with increasing DR. Once on the inertial domain, self-similar behaviors occur, but modified (sometimes subtly) by the modified distribution of characteristic y-scaling behavior of the Reynolds stress motions.

Thermo-mechanical actuator-based miniature tagging module for localization in capsule endoscopy

NASA Astrophysics Data System (ADS)

Chandrappan, Jayakrishnan; Ruiqi, Lim; Su, Nandar; Yen Yi, Germaine Hoe; Vaidyanathan, Kripesh

2011-04-01

Capsule endoscopy is a frontline medical diagnostic tool for the gastro intestinal tract disorders. During diagnosis, efficient localization techniques are essential to specify a pathological area that may require further diagnosis or treatment. This paper presents the development of a miniature tagging module that relies on a novel concept to label the region of interest and has the potential to integrate with a capsule endoscope. The tagging module is a compact thermo-mechanical actuator loaded with a biocompatible micro tag. A low power microheater attached to the module serves as the thermal igniter for the mechanical actuator. At optimum temperature, the actuator releases the micro tag instantly and penetrates the mucosa layer of a GI tract, region of interest. Ex vivo animal trials are conducted to verify the feasibility of the tagging module concept. X-ray imaging is used to detect the location of the micro tag embedded in the GI tract wall. The method is successful, and radiopaque micro tags can provide valuable pre-operative position information on the infected area to facilitate further clinical procedures.

Tagging studies of mule deer fawns on the Hanford Site, 1969 to 1977

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

Eberhardt, L.E.; Hedlund, J.D.; Rickard, W.H.

1979-10-01

From 1969 to 1977, 346 mule deer (Odocoileus hemionus) fawns were tagged and released on islands and shoreline habitat associated with the Columbia River on the Hanford Site in south-central Washington. The purpose was to determine the movement of mule deer along the Columbia River shoreline from the Hanford Site through tag recovery. Twenty-one tagged deer have been killed primarily by hunters near the Hanford Site or on areas of the Hanford Site open to public access. Movements of up to 113 km from Hanford have been documented. Although the Columbia River at Hanford is one of the largest andmore » most swift-flowing rivers in North America it is not an impassable barrier to mule deer. River islands are important and perhaps critical fawining habitat for the local deer herd. The selection of these islands by pregnant female deer is apparently influenced by predation, human access, and recreational use of islands. The number of fawns captured decreased during the latter years of the study (1974 to 1977). This is probably a reflection of an actual decrease in deer productivity, particularly along the upper stretch of the Columbia flowing through the Hanford Site. The reasons for this apparent decrease are unkown.« less

Evaluation of strobe lights to reduce turbine entrainment of juvenile steelhead (Oncorhynchus mykiss) at Cowlitz Falls Dam, Washington

USGS Publications Warehouse

Kock, Tobias J.; Evans, Scott D.; Liedtke, Theresa L.; Rondorf, Dennis W.; Kohn, Mike

2009-01-01

We conducted a radiotelemetry evaluation to determine if strobe lights could be used to decrease turbine entrainment of juvenile steelhead (Oncorhynchus mykiss) at Cowlitz Falls Dam, Washington. We found that radio-tagged juvenile steelhead approached and entered two spillbays (one lighted, one unlighted) in equal proportions. However, the presence of strobe lights was associated with decreased spillbay residence time of juvenile steelhead and increased passage through induction slots (secondary turbine intakes located upstream of the ogee on the spillway). Mean residence time of tagged fish inside the lighted spillbay was 14 min compared to 62 min inside the unlighted spillbay. Radio-tagged steelhead passed through induction slots at a higher proportion in the lighted spillbay (55%) than in the unlighted spillbay (26%). Recent studies have suggested that strobe lights can induce torpor in juvenile salmonids. We believe that strobe light exposure affected fish in our study at a location where they were susceptible to high flows thereby reducing mean residence time and increasing the proportion of tagged fish entering induction slots in the lighted spillbay. Our results suggest that factors such as deployment location, exposure, and flow are important variables that should be considered when evaluating strobe lights as a potential fish-deterring management tool.

Highly multiparametric analysis by mass cytometry.

PubMed

Ornatsky, Olga; Bandura, Dmitry; Baranov, Vladimir; Nitz, Mark; Winnik, Mitchell A; Tanner, Scott

2010-09-30

This review paper describes a new technology, mass cytometry, that addresses applications typically run by flow cytometer analyzers, but extends the capability to highly multiparametric analysis. The detection technology is based on atomic mass spectrometry. It offers quantitation, specificity and dynamic range of mass spectrometry in a format that is familiar to flow cytometry practitioners. The mass cytometer does not require compensation, allowing the application of statistical techniques; this has been impossible given the constraints of fluorescence noise with traditional cytometry instruments. Instead of "colors" the mass cytometer "reads" the stable isotope tags attached to antibodies using metal-chelating labeling reagents. Because there are many available stable isotopes, and the mass spectrometer provides exquisite resolution between detection channels, many parameters can be measured as easily as one. For example, in a single tube the technique allows for the ready detection and characterization of the major cell subsets in blood or bone marrow. Here we describe mass cytometric immunophenotyping of human leukemia cell lines and leukemia patient samples, differential cell analysis of normal peripheral and umbilical cord blood; intracellular protein identification and metal-encoded bead arrays. Copyright © 2010 Elsevier B.V. All rights reserved.

ezTag: tagging biomedical concepts via interactive learning.

PubMed

Kwon, Dongseop; Kim, Sun; Wei, Chih-Hsuan; Leaman, Robert; Lu, Zhiyong

2018-05-18

Recently, advanced text-mining techniques have been shown to speed up manual data curation by providing human annotators with automated pre-annotations generated by rules or machine learning models. Due to the limited training data available, however, current annotation systems primarily focus only on common concept types such as genes or diseases. To support annotating a wide variety of biological concepts with or without pre-existing training data, we developed ezTag, a web-based annotation tool that allows curators to perform annotation and provide training data with humans in the loop. ezTag supports both abstracts in PubMed and full-text articles in PubMed Central. It also provides lexicon-based concept tagging as well as the state-of-the-art pre-trained taggers such as TaggerOne, GNormPlus and tmVar. ezTag is freely available at http://eztag.bioqrator.org.

Immobilization of proteins onto microbeads using a DNA binding tag for enzymatic assays.

PubMed

Kojima, Takaaki; Mizoguchi, Takuro; Ota, Eri; Hata, Jumpei; Homma, Keisuke; Zhu, Bo; Hitomi, Kiyotaka; Nakano, Hideo

2016-02-01

A novel DNA-binding protein tag, scCro-tag, which is a single-chain derivative of the bacteriophage lambda Cro repressor, has been developed to immobilize proteins of interest (POI) on a solid support through binding OR consensus DNA (ORC) that is tightly bound by the scCro protein. The scCro-tag successfully bound a transglutaminase 2 (TGase 2) substrate and manganese peroxidase (MnP) to microbeads via scaffolding DNA. The resulting protein-coated microbeads can be utilized for functional analysis of the enzymatic activity using flow cytometry. The quantity of bead-bound proteins can be enhanced by increasing the number of ORCs. In addition, proteins with the scCro-tag that were synthesized using a cell-free protein synthesis system were also immobilized onto the beads, thus indicating that this bead-based system would be applicable to high-throughput analysis of various enzymatic activities. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

LlamaTags: A Versatile Tool to Image Transcription Factor Dynamics in Live Embryos.

PubMed

Bothma, Jacques P; Norstad, Matthew R; Alamos, Simon; Garcia, Hernan G

2018-06-14

Embryonic cell fates are defined by transcription factors that are rapidly deployed, yet attempts to visualize these factors in vivo often fail because of slow fluorescent protein maturation. Here, we pioneer a protein tag, LlamaTag, which circumvents this maturation limit by binding mature fluorescent proteins, making it possible to visualize transcription factor concentration dynamics in live embryos. Implementing this approach in the fruit fly Drosophila melanogaster, we discovered stochastic bursts in the concentration of transcription factors that are correlated with bursts in transcription. We further used LlamaTags to show that the concentration of protein in a given nucleus heavily depends on transcription of that gene in neighboring nuclei; we speculate that this inter-nuclear signaling is an important mechanism for coordinating gene expression to delineate straight and sharp boundaries of gene expression. Thus, LlamaTags now make it possible to visualize the flow of information along the central dogma in live embryos. Copyright © 2018 Elsevier Inc. All rights reserved.

Fabrication of Magnetic Barcoded Microcarriers for Biomolecular Labeling: SU-8 Encapsulated Magnetic Tags

NASA Astrophysics Data System (ADS)

Palfreyman, Justin J.; Beldon, Patrick; Hong, Bingyan; Vyas, Kunal N.; Cooper, Joshaniel F. K.; Mitrelias, Thanos; Barnes, Crispin H. W.

2010-12-01

Rows of rectangular magnetic elements with different aspect ratio are encapsulated in polymer microcarriers to form a novel magnetic label, or tag, for multiplexed biological and chemical assays. We demonstrate that each tag can be encoded using an external magnetic field applied to the whole tag, which will allow for in-flow writing, thanks to shape-anisotropy controlled coercivity of the individual bits. This paper focuses on the fabrication of our 2nd generation tags, which facilitate optical trapping, do not require a sacrificial release layer, and the alignment procedure has been simplified to a single step. A new procedure is described for recovering a functional surface from fully cross-linked SU-8 via a cerium (IV) ammonium nitrate based chemical etch, and a novel method for releasing patterned photoresist from a bare Si wafer is discussed. In addition, a series of homobifunctional amine spacer compounds are compared as a method of increasing the binding efficiency of surface probe molecules.

Electrochemically Initiated Tagging of Thiols Using an Electrospray Ionization-Based Liquid Microjunction Surface Sampling Probe Two-Electrode Cell

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

Van Berkel, Gary J; Kertesz, Vilmos

2009-01-01

This paper reports on the conversion of a liquid microjunction surface sampling probe (LMJ-SSP) into a two electrode electrochemical cell using a conductive sample surface and the probe as the two electrodes with an appropriate battery powered circuit. With this LMJ-SSP, two-electrode cell arrangement, tagging of analyte thiol functionalities (in this case peptide cysteine residues) with hydroquinone tags was initiated electrochemically using a hydroquinone doped solution when the analyte either was initially in solution or was sampled from a surface. Efficient tagging (~90%), at flow rates of 5-10 L/min, could be achieved for up to at least two cysteines onmore » a peptide. The high tagging efficiency observed was explained with a simple kinetic model. In general, the incorporation of a two-electrode electrochemical cell, or other multiple electrode arrangement, into the LMJ-SSP is expected to add to the versatility of this approach for surface sampling and ionization coupled with mass spectrometric detection.« less

Performance of Encounternet Tags: Field Tests of Miniaturized Proximity Loggers for Use on Small Birds

PubMed Central

Levin, Iris I.; Zonana, David M.; Burt, John M.; Safran, Rebecca J.

2015-01-01

Proximity logging is a new tool for understanding social behavior as it allows for accurate quantification of social networks. We report results from field calibration and deployment tests of miniaturized proximity tags (Encounternet), digital transceivers that log encounters between tagged individuals. We examined radio signal behavior in relation to tag attachment (tag, tag on bird, tag on saline-filled balloon) to understand how radio signal strength is affected by the tag mounting technique used for calibration tests. We investigated inter-tag and inter-receiver station variability, and in each calibration test we accounted for the effects of antennae orientation. Additionally, we used data from a live deployment on breeding barn swallows (Hirundo rustica erythrogaster) to analyze the quality of the logs, including reciprocal agreement in dyadic logs. We evaluated the impact (in terms of mass changes) of tag attachment on the birds. We were able to statistically distinguish between RSSI values associated with different close-proximity (<5m) tag-tag distances regardless of antennae orientation. Inter-tag variability was low, but we did find significant inter-receiver station variability. Reciprocal agreement of dyadic logs was high and social networks were constructed from proximity tag logs based on two different RSSI thresholds. There was no evidence of significant mass loss in the time birds were wearing tags. We conclude that proximity loggers are accurate and effective for quantifying social behavior. However, because RSSI and distance cannot be perfectly resolved, data from proximity loggers are most appropriate for comparing networks based on specific RSSI thresholds. The Encounternet system is flexible and customizable, and tags are now light enough for use on small animals (<50g). PMID:26348329

Behavioral and reproductive effects of bird-borne data logger attachment on Brown Pelicans (Pelecanus occidentalis) on three temporal scales

USGS Publications Warehouse

Lamb, Juliet S.; Satgé, Yvan G.; Fiorello, Christine V.; Jodice, Patrick G. R.

2017-01-01

Although the use of bird-borne data loggers has become widespread in avian field research, the effects of capture and transmitter attachment on behavior and demographic rates are not often measured. Tag- and capture-induced effects on individual behavior, survival and reproduction may limit extrapolation of transmitter data to wider populations. However, measuring individual responses to capture and tagging is a necessary step in developing research techniques that minimize negative effects. We measured the short-term behavioral effects of handling and GPS transmitter attachment on Brown Pelicans under both captive and field conditions, and followed tagged individuals through a full breeding season to assess whether capture and transmitter attachment increased rates of nest abandonment or breeding failure. We observed slight increases in preening among tagged individuals 0–2 h after capture relative to controls that had not been captured or tagged, with a corresponding reduction in time spent resting. One to three days post-capture, nesting behavior of tagged pelicans resembled that of neighbors that had not been captured or tagged. Eighty-eight percent of tagged breeders remained at the same nest location for more than 48 h after capture, attending nests and chicks for an average of 49 days, and 51% were assumed to successfully fledge young. Breeding success was driven primarily by variation in location; however, sex and handling time also influenced the probability of successful breeding in tagged pelicans, suggesting that individual characteristics and the capture process itself can confound the effects of capture and transmitter attachment. We conclude that pelicans fitted with GPS transmitters exhibit comparable behaviors to untagged individuals within a day of capture and that GPS tracking is a viable technique for studying behavior and demography in this species. We also identify measures to minimize post-capture nest abandonment rates in tracking studies, including minimizing handling time and covering nests during processing.

Preparative SDS PAGE as an Alternative to His-Tag Purification of Recombinant Amelogenin

PubMed Central

Gabe, Claire M.; Brookes, Steven J.; Kirkham, Jennifer

2017-01-01

Recombinant protein technology provides an invaluable source of proteins for use in structure-function studies, as immunogens, and in the development of therapeutics. Recombinant proteins are typically engineered with “tags” that allow the protein to be purified from crude host cell extracts using affinity based chromatography techniques. Amelogenin is the principal component of the developing enamel matrix and a frequent focus for biomineralization researchers. Several groups have reported the successful production of recombinant amelogenins but the production of recombinant amelogenin free of any tags, and at single band purity on silver stained SDS PAGE is technically challenging. This is important, as rigorous structure-function research frequently demands a high degree of protein purity and fidelity of protein sequence. Our aim was to generate His-tagged recombinant amelogenin at single band purity on silver stained SDS PAGE for use in functionality studies after His-tag cleavage. An acetic acid extraction technique (previously reported to produce recombinant amelogenin at 95% purity directly from E. coli) followed by repeated rounds of nickel column affinity chromatography, failed to generate recombinant amelogenin at single band purity. This was because following an initial round of nickel column affinity chromatography, subsequent cleavage of the His-tag was not 100% efficient. A second round of nickel column affinity chromatography, used in attempts to separate the cleaved His-tag free recombinant from uncleaved His-tagged contaminants, was still unsatisfactory as cleaved recombinant amelogenin exhibited significant affinity for the nickel column. To solve this problem, we used preparative SDS PAGE to successfully purify cleaved recombinant amelogenins to single band purity on silver stained SDS PAGE. The resolving power of preparative SDS PAGE was such that His-tag based purification of recombinant amelogenin becomes redundant. We suggest that acetic acid extraction of recombinant amelogenin and subsequent purification using preparative SDS PAGE provides a simple route to highly purified His-tag free amelogenin for use in structure-function experiments and beyond. PMID:28670287

Development and evaluation of a boat-mounted RFID antenna for monitoring freshwater mussels

USGS Publications Warehouse

Fischer, Jesse R.; Neebling, Travis E.; Quist, Michael C.

2012-01-01

Development of radio frequency identification (RFID) technology and passive integrated transponder (PIT) tags has substantially increased the ability of researchers and managers to monitor populations of aquatic organisms. However, use of transportable RFID antenna systems (i.e., backpack-mounted) is currently limited to wadeable aquatic environments (<1.4 m water depth). We describe the design, construction, and evaluation of a boat-mounted RFID antenna to detect individually PIT-tagged benthic aquatic organisms (mussels). We evaluated the effects of tag orientation on detection distances in water with a 32-mm half-duplex PIT tag. Detection distances up to 50 cm from the antenna coils were obtained, but detection distance was dependent on tag orientation. We also evaluated detection distance of PIT tags beneath the sediment to simulate detection of burrowing mussels with 23- and 32-mm tags. In sand substrate, the maximum detection distance varied from 3.5 cm and 4.5 cm (vertical tag orientation) to 24.7 cm and 39.4 cm (45° tag orientation) for the 23- and 32-mm PIT tags, respectively. Our results suggest a 1.4-m total detection width for tagged mussels on the substrate surface by the boat-mounted antenna system regardless of tag orientation. However, burrowed mussels may require multiple passes to increase detection that would be influenced by depth, tag orientation, and tag size. Construction of the boat-mounted antenna was relatively low in cost (<500 USD) and had several advantages (less labor and time intensive, increased safety) over traditional mussel sampling techniques (diving, snorkeling) in nonwadeable habitats.

Using passive integrated transponder (PIT) systems for terrestrial detection of blue-spotted salamanders (Ambystoma laterale) in situ

USGS Publications Warehouse

Ryan, Kevin J.; Zydlewski, Joseph D.; Calhoun, Aram J.K.

2014-01-01

Pure-diploid Blue-spotted Salamanders (Ambystoma laterale) are the smallest members of the family Ambystomatidae which makes tracking with radio-transmitters difficult because of small battery capacity. Passive integrated transponder (PIT) tags provide another tracking approach for small fossorial animals such as salamanders. We evaluated the use of portable PIT tag readers (PIT packs) to detect PIT tag-implanted pure-diploid Blue-spotted Salamanders in situ. We also examined the detection probability of salamanders with PIT tags held in enclosures in wetland and terrestrial habitats, as well as the underground detection range of PIT packs by scanning for buried tags not implanted into salamanders. Of the 532 PIT tagged salamanders, we detected 6.84% at least once during scanning surveys. We scanned systematically within a 13.37 ha area surrounding a salamander breeding pool on 34 occasions (~119 hours of survey time) and detected PIT tags 74 times. We detected 55% of PITs in tagged salamanders and 45%were expelled tags. We were able to reliably detect buried PIT tags from 1–22cm below the ground surface. Because nearly half the locations represented expelled tags, our data suggest this technique is inappropriate for future studies of pure-diploid Blue-spotted Salamanders, although it may be suitable for polyploid Blue-spotted Salamanders and other ambystomatid species, which are larger in size and may exhibit higher tag retention rates. It may also be prudent to conduct long-term tag retention studies in captivity before tagging and releasing salamanders for in situ study, and to double-mark individuals.

Fluorescent labeling of SNAP-tagged proteins in cells.

PubMed

Lukinavičius, Gražvydas; Reymond, Luc; Johnsson, Kai

2015-01-01

One of the most prominent self-labeling tags is SNAP-tag. It is an in vitro evolution product of the human DNA repair protein O (6)-alkylguanine-DNA alkyltransferase (hAGT) that reacts specifically with benzylguanine (BG) and benzylchloropyrimidine (CP) derivatives, leading to covalent labeling of SNAP-tag with a synthetic probe (Gronemeyer et al., Protein Eng Des Sel 19:309-316, 2006; Curr Opin Biotechnol 16:453-458, 2005; Keppler et al., Nat Biotechnol 21:86-89, 2003; Proc Natl Acad Sci U S A 101:9955-9959, 2004). SNAP-tag is well suited for the analysis and quantification of fused target protein using fluorescence microscopy techniques. It provides a simple, robust, and versatile approach to the imaging of fusion proteins under a wide range of experimental conditions.

Enhanced UHF RFID tags for drug tracing.

PubMed

Catarinucci, Luca; Colella, Riccardo; De Blasi, Mario; Patrono, Luigi; Tarricone, Luciano

2012-12-01

Radio Frequency Identification (RFID) technology is playing a crucial role for item-level tracing systems in healthcare scenarios. The pharmaceutical supply chain is a fascinating application context, where RFID can guarantee transparency in the drug flow, supporting both suppliers and consumers against the growing counterfeiting problem. In such a context, the choice of the most adequate RFID tag, in terms of shape, frequency, size and reading range, is crucial. The potential presence of items containing materials hostile to the electromagnetic propagation exasperates the problem. In addition, the peculiarities of the different RFID-based checkpoints make even more stringent the requirements for the tag. In this work, the performance of several commercial UHF RFID tags in each step of the pharmaceutical supply chain has been evaluated, confirming the expected criticality. On such basis, a guideline for the electromagnetic design of new high-performance tags capable to overcome such criticalities has been defined. Finally, driven by such guidelines, a new enhanced tag has been designed, realized and tested. Due to patent pending issues, the antenna shape is not shown. Nevertheless, the optimal obtained results do not lose their validity. Indeed, on the one hand they demonstrate that high performance item level tracing systems can actually be implemented also in critical operating conditions. On the other hand, they encourage the tag designer to follow the identified guidelines so to realize enhanced UHF tags.

Development of Improved Satellite-Linked Transmitters, Physiological Recorders, and Attachment Techniques for Monitoring Beaked Whales

DTIC Science & Technology

2010-09-01

our previous petals by 20% in order to minimize tissue damage upon entry while still maintaining enough retention power. We switched to using 0.020...whales in Southern California, and 5 LIMPET Mk10-A tags to sperm whales in Southeast Alaska. As of 30 September 2010, 1 of the Cuvier’s tags is still...sending dive depth data (94 days after attachment) and 3 of the sperm whale tags are still transmitting dive data (46 days after attachment

Evaluation of Ginger (Zingiber officinale Roscoe) Bioactive Compounds in Increasing the Ratio of T-cell Surface Molecules of CD3+CD4+:CD3+CD8+ In-Vitro.

PubMed

Tejasari, Dr

2007-09-01

The potential ability of ginger bioactive compounds in increasing the ratio of T-cell surface molecules of CD3+CD4+:CD3+CD8+ was investigated using dual tagging FITC and PE of monoclonal antibody anti-human with its fluorescence measured by flow cytometer. Oleoresin was extracted using sinkhole distillation technique. Its components namely, gingerol in fraction-1, shogaol in fraction 2 and zingeron in fraction-3 were separated by column vacuum chromatography method. The doses of oleoresin, gingerol, shogaol, and zingeron tested were 50, 100,150, 200, and 250 μg/ml. Lymphocytes (2x106 cell/ml) from human peripheral blood were isolated using ficoll density gradient technique, and cultured in the presence of the compounds in RPMI-1640 medium and phytohemaglutinin (PHA) mitogen for 96 h under normal conditions. Percentages of T-cell surface molecules (CD4+ and CD8+) were determined using dual-tagging FITC and PE fluorescents labeled on monoclonal antibody anti human. The fluorescence-labeled bands on the T-cell surface molecules were counted using flow cytometer. The experiment revealed that oleoresin and its three fractions increased the percentage of CD3+CD4+. The compound in fraction 3 of oleoresin at 200 μg/ml increased by the highest percentage of CD3+CD4+ of 9%, but slightly decreased the percentage of CD3+CD8+. These ginger bioactive compounds increased the ratio of CD3+CD4:CD3+CD8+ T-cells with the highest increment of 30% from effects of 200 μg/ml fraction 3 of oleoresin. This in vitro finding revealed that ginger bioactive compounds potentially increased cellular and humoral immune response. Further clinical studies are needed to confirm the benefits of these ginger bioactive compounds as a potential functional food for testing on HIV infected patients.

Flow-Tagging Velocimetry for Hypersonic Flows Using Fluorescence of Nitric Oxide

NASA Technical Reports Server (NTRS)

Danehy, Paul M.; OByrne, Sean; Houwing, A. Frank P.; Fox, Jodie S.; Smith, Daniel R.

2003-01-01

We demonstrate a new variation of molecular-tagging velocimetry for hypersonic flows based on laser-induced fluorescence. A thin line of nitric-oxide molecules is excited with a laser beam and then, after a time delay, a fluorescence image of the displaced line is acquired. One component of velocity is determined from the time of flight. This method is applied to measure the velocity profile in a Mach 8.5 laminar, hypersonic boundary layer in the Australian National University s T2 free-piston shock tunnel. The single-shot velocity measurement uncertainty in the freestream was found to be 3.5%, based on 90% confidence. The method is also demonstrated in the separated flow region forward of a blunt fin attached to a flat plate in a Mach 7.4 flow produced by the Australian National University s T3 free-piston shock tunnel. The measurement uncertainty in the blunt fin experiment is approximately 30%, owing mainly to low fluorescence intensities, which could be improved significantly in future experiments. This velocimetry method is applicable to very high-speed flows that have low collisional quenching of the fluorescing species. It is particularly convenient in facilities where planar laser-induced fluorescence is already being performed.

Snake River fall Chinook salmon life history investigations, annual report 2008

USGS Publications Warehouse

Tiffan, Kenneth F.; Connor, William P.; Bellgraph, Brian J.; Buchanan, Rebecca A.

2010-01-01

In 2009, we used radio and acoustic telemetry to evaluate the migratory behavior, survival, mortality, and delay of subyearling fall Chinook salmon in the Clearwater River and Lower Granite Reservoir. We released a total of 1,000 tagged hatchery subyearlings at Cherry Lane on the Clearwater River in mid August and we monitored them as they passed downstream through various river and reservoir reaches. Survival through the free-flowing river was high (>0.85) for both radio- and acoustic-tagged fish, but dropped substantially as fish delayed in the Transition Zone and Confluence areas. Estimates of the joint probability of migration and survival through the Transition Zone and Confluence reaches combined were similar for both radio- and acoustic-tagged fish, and ranged from about 0.30 to 0.35. Estimates of the joint probability of delaying and surviving in the combined Transition Zone and Confluence peaked at the beginning of the study, ranging from 0.323 ( SE =NA; radio-telemetry data) to 0.466 ( SE =0.024; acoustic-telemetry data), and then steadily declined throughout the remainder of the study. By the end of October, no live tagged juvenile salmon were detected in either the Transition Zone or the Confluence. As estimates of the probability of delay decreased throughout the study, estimates of the probability of mortality increased, as evidenced by the survival estimate of 0.650 ( SE =0.025) at the end of October (acoustic-telemetry data). Few fish were detected at Lower Granite Dam during our study and even fewer fish passed the dam before PIT-tag monitoring ended at the end of October. Five acoustic-tagged fish passed Lower Granite Dam in October and 12 passed the dam in November based on detections in the dam tailrace; however, too few detections were available to calculate the joint probabilities of migrating and surviving or delaying and surviving. Estimates of the joint probability of migrating and surviving through the reservoir was less than 0.2 based on acoustic-tagged fish. Migration rates of tagged fish were highest in the free-flowing river (median range = 36 to 43 km/d) but were generally less than 6 km/d in the reservoir reaches. In particular, median migration rates of radio-tagged fish through the Transition Zone and Confluence were 3.4 and 5.2 km/d, respectively. Median migration rate for acoustic-tagged fish though the Transition Zone and Confluence combined was 1 km/d.

Improved fatty acid analysis of conjugated linoleic acid rich egg yolk triacylglycerols and phospholipid species.

PubMed

Shinn, Sara; Liyanage, Rohana; Lay, Jack; Proctor, Andrew

2014-07-16

Reports from chicken conjugated linoleic acid (CLA) feeding trials are limited to yolk total fatty acid composition, which consistently described increased saturated fatty acids and decreased monounsaturated fatty acids. However, information on CLA triacylglycerol (TAG) and phospholipid (PL) species is limited. This study determined the fatty acid composition of total lipids in CLA-rich egg yolk produced with CLA-rich soy oil, relative to control yolks using gas chromatography with flame ionization detection (GC-FID), determined TAG and PL fatty acid compositions by thin-layer chromatography-GC-FID (TLC-GC-FID), identified intact PL and TAG species by TLC-matrix-assisted laser desorption/ionization mass spectrometry (TLC-MALDI-MS), and determined the composition of TAG and PL species in CLA and control yolks by direct flow infusion electrospray ionization MS (DFI ESI-MS). In total, 2 lyso-phosphatidyl choline (LPC) species, 1 sphingomyelin species, 17 phosphatidyl choline species, 19 TAG species, and 9 phosphatidyl ethanolamine species were identified. Fifty percent of CLA was found in TAG, occurring predominantly in C52:5 and C52:4 TAG species. CLA-rich yolks contained significantly more LPC than did control eggs. Comprehensive lipid profiling may provide insight on relationships between lipid composition and the functional properties of CLA-rich eggs.

Survival and growth of juvenile Pacific lampreys tagged with passive integrated transponders (PIT) in freshwater and seawater

USGS Publications Warehouse

Mesa, Matthew G.; Copeland, Elizabeth S.; Christiansen, Helena E.; Gregg, Jacob L.; Roon, Sean R.; Hershberger, Paul K.

2012-01-01

Tagging methods are needed for both adult and juvenile life stages of Pacific lampreys Lampetra tridentata to better understand their biology and factors contributing to their decline. We developed a safe and efficient technique for tagging juvenile Pacific lampreys with passive integrated transponder (PIT) tags. We tested the short-term survival of PIT-tagged juvenile lampreys in freshwater at four temperatures (9, 12, 15, and 18°C) and their long-term growth and survival in seawater. For both experiments there was little to no tag loss, and juvenile lampreys in freshwater showed high survival at all temperatures at 7 d (95–100%) and 14 d (88–100%) posttagging. Prolonged holding (40 d) resulted in significantly lower survival (28–79%) at warmer temperatures (12–18°C). For juvenile lampreys tagged in freshwater and then transitioned to seawater, survival was 97% for tagged fish until day 94, and at the end of 6 months, survival was about 58% for both tagged and control fish. About half of the tagged and control fish that survived in seawater grew, but there was no difference in growth between the two groups. In freshwater, but not in seawater, most fish that died had an aquatic fungal infection. In both experiments, survival increased with increasing fish length at tagging. Our results indicate that tags similar in size to a 9-mm PIT tag are a feasible option for tagging metamorphosed juvenile lampreys migrating downstream and that when fungal infections are mitigated—as in seawater—long-term (at least 6 months) survival of tagged juvenile lampreys is high.

Sediment tracing from small torrential channels to gravel-bed rivers using pit tags method. A case study from the upper Guil catchment.

NASA Astrophysics Data System (ADS)

Graff, Kévin; Viel, Vincent; Carlier, Benoit; Lissak, Candide; Arnaud-Fassetta, Gilles; Fort, Monique; Madelin, Malika

2016-04-01

In mountainous areas, especially in large catchments with torrential tributaries, the production and sediment transport significantly increase flood impacts in the valley bottoms. The quantification and characterisation of sedimentary transfers are therefore major challenges to provide better flood risk management. As a part of SAMCO (ANR 12 SENV-0004 SAMCO) project, for mountain hazard assessment in a context of global changes, we tried to improve the knowledge of these hydromorphological systems at both spatial and temporal scales, by identifying sediment supply and sediment dynamics from torrential tributaries to the main channel. A sediment budget was used as a tool for quantifying erosion, transport and deposition processes. This research is focused on the upper Guil catchment (Queyras, Southern French Alps - 317 km2) entrenched in "schistes lustrés" and ophiolitic bedrock. This catchment is prone to catastrophic summer floods [June 1957 (>R.I. 100 yr), June 2000 (R.I. 30 yr)] characterised by huge sediment transport from tributaries to downvalley, very much facilitated by strong hillslope-channel connectivity (about 12,000 m3 volume of sediment aggraded in the Peyronnelle fan during the June 2000 RI-30 year flood event). We intend to highlight sediment dynamics on small torrential channels and its connection with gravel-bed streams. Four study sites characterised by avalanche and debris flow-dominated channels located in the upper Guil catchment were investigated. In order to better assess sediment movement, we used the pit-tags technique. In total, 560 pit-tags (pt) have been implemented in four catchments: Peyronnelle (320pt), Combe Morel (40pt), Bouchouse (120pt), and Maloqueste (80pt). Distances and trajectories of gravels sediments have been monitored since two years during summer periods. We specifically describe results obtained along the Peyronnelle channel affected by a large debris-flow during august 2015. Data are used to discuss lag time, processes and thresholds needed to observe significant sediments fluxes. Results highlight the pulsating character of sediment fluxes associated with high magnitude and low frequency events and indicate the strongest functionality of debris flow-dominated channels. We intend to continue this monitoring long enough to observe sediment connection with gravel-bed streams.

Precision of FLEET Velocimetry Using High-speed CMOS Camera Systems

NASA Technical Reports Server (NTRS)

Peters, Christopher J.; Danehy, Paul M.; Bathel, Brett F.; Jiang, Naibo; Calvert, Nathan D.; Miles, Richard B.

2015-01-01

Femtosecond laser electronic excitation tagging (FLEET) is an optical measurement technique that permits quantitative velocimetry of unseeded air or nitrogen using a single laser and a single camera. In this paper, we seek to determine the fundamental precision of the FLEET technique using high-speed complementary metal-oxide semiconductor (CMOS) cameras. Also, we compare the performance of several different high-speed CMOS camera systems for acquiring FLEET velocimetry data in air and nitrogen free-jet flows. The precision was defined as the standard deviation of a set of several hundred single-shot velocity measurements. Methods of enhancing the precision of the measurement were explored such as digital binning (similar in concept to on-sensor binning, but done in post-processing), row-wise digital binning of the signal in adjacent pixels and increasing the time delay between successive exposures. These techniques generally improved precision; however, binning provided the greatest improvement to the un-intensified camera systems which had low signal-to-noise ratio. When binning row-wise by 8 pixels (about the thickness of the tagged region) and using an inter-frame delay of 65 micro sec, precisions of 0.5 m/s in air and 0.2 m/s in nitrogen were achieved. The camera comparison included a pco.dimax HD, a LaVision Imager scientific CMOS (sCMOS) and a Photron FASTCAM SA-X2, along with a two-stage LaVision High Speed IRO intensifier. Excluding the LaVision Imager sCMOS, the cameras were tested with and without intensification and with both short and long inter-frame delays. Use of intensification and longer inter-frame delay generally improved precision. Overall, the Photron FASTCAM SA-X2 exhibited the best performance in terms of greatest precision and highest signal-to-noise ratio primarily because it had the largest pixels.

Enabling Technologies for Microfluidic Flow Control and Detection

NASA Astrophysics Data System (ADS)

Leslie, Daniel Christopher

Advances in microfluidic technologies have expanded conventional chemical and biological techniques to the point where we can envision rapid, inexpensive and portable analysis. Among the numerous challenges in the development of portable, chip-based technologies are simple flow control and detection strategies, which will be essential to widespread acceptance and implementation at both the point-of-care and in locales with limited facilities/resources. The research presented in this dissertation is focused on the development of precise flow control techniques and new, simplified detection technologies aimed at addressing these challenges. An introduction to the concepts important to microfluidics and a brief history to the field are presented in Chapter 1. Chapter 2 will present the development of a technique for the precise control of small volumes of liquids, where well-studied electrical circuit concepts are employed to create frequency-dependent microfluidic circuits. In this system, elastomeric thin films act as fluidic capacitors and diodes, which, when combined with resistors (channels), make fluidic circuits that are described by analytical models. Metering of two separate chemical inputs with a single oscillatory pneumatic control line is demonstrated by combining simple fluidic circuits (i.e., band-pass filters) to significantly reduce the external hardware required for microfluidic flow control. In order to quantify multiple flow profiles in microfluidic circuits, a novel multiplexed flow measurement method using visible dyes is introduced in Chapter 3 and rapidly determines individual flow in connected channels, post-fabrication device quality and solution viscosity. Another thrust of this dissertation research has been to develop miniaturized bioanalytical systems. Chapter 4 describes the adaption of a nucleic-acid-tagged antibody protein detection reaction to a microfluidic platform for detection of down to 5 E. coli O157:H7 cells. Furthermore, a completely non-contact temperature control platform is developed in Chapter 5 for forensic human identification reactions, based on interferometric temperature sensing and infrared-mediated heating, which simplifies the microfluidic device and its operation. Finally, possible future directions are outlined in Chapter 6 including further simplification of microfluidic instrumentation.

Lower Granite Dam Smolt Monitoring Program, 2003-2004 Annual Report.

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

Mensik, Fred; Rapp, Shawn; Ross, Doug

2004-08-01

The 2003 fish collection season at Lower Granite Dam Juvenile Fish Facility (LGR) was characterized by water temperatures, total flows and spill that were below the five year average, low levels of debris, and increased smolt collection numbers compared to 2002 with the exception of unclipped sockeye/kokanee. There were 6,183,825 juvenile salmonids collected. Of these, 6,054,167 were transported to release sites below Bonneville Dam, 5,957,885 by barge and 96,282 by truck. An additional 102,340 fish were bypassed back to the river, primarily due to research projects with another 62,122 bypassed through the PIT-tag bypass system. According to the PTAGIS database,more » 152,268 PIT-tagged fish were detected at Lower Granite Dam. Of these, Smolt Monitoring Staff recorded 345 PIT-tagged raceway and sample mortalities. Of the 6,183,825 total fish collected, 113,290 were PIT-tagged or radio tagged and 380 were sacrificed by researchers. The collection included 836,885 fish that had hatchery marks other than clipped fins (elastomer, freeze brands or Coded Wire Tags). An estimated 54,857 incidental fish were collected with an additional 8,730 adult salmonids removed from the separator.« less

Evaluation of Fish Movements, Migration Patterns, and Population Abundance with Streamwidth PIT Tag Interrogation Systems, Final Report 2002.

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

Zydlewski, Gayle; Winter, Christiane; McClanahan, Dee

2003-02-01

Two remote Streamwidth PIT tag Interrogation systems (SPIs) were operated continuously for over one year to test the feasibility of these systems for generating movement, migration, survival and smolt production estimates for salmonids. A total of 1,588 juvenile (< 100 mm FL) naturally produced salmonids (7 coho salmon, 482 cutthroat trout, and 1,099 steelhead) were PIT tagged above the upstream-most SPI (9 sites approximately 1 linear km each) in Fall 2001. Age at tagging for wild caught cutthroat and steelhead was 1 year. SPIs were operating before any PIT tagged fish were released in the creek. Over 390,000 detections weremore » recorded from October 2001 to 31 July 2002. Efficiencies were site dependent, but overall detection efficiency for the creek was 97% with 95% confidence intervals of 91-100%. PIT tag detection efficiency ranged from 55-100% depending on the SPI and varied throughout the year with average efficiencies of 73% and 89%. SPI efficiency of PIT tag detection was not completely dependent on electronics noise levels or environmental conditions. Fish from all tagging locations were detected at the SPIs. Steelhead and cutthroat trout were primarily detected moving in the Spring (April-June) coincident with the anticipated smolt migration. Steelhead were also detected moving past SPIs at lower numbers in the Fall and Winter. Travel time between SPIs (downstream movement) was highly dependent on time of year. Travel time in the Spring was significantly faster (34.4 {+-} 7.0 hours) for all species than during any other time of year (763.1 {+-} 267.0 hours). Steelhead and cutthroat migrating in the Spring were the same age as those that did not migrate in the Spring. Peak of steelhead migration recorded at the two SPIs was 5/11 and 5/12 and the peak in the screw trap was recorded on 5/17. Steelhead smolt production estimates using SPIs (3,802 with 95% confidence intervals of 3,440 - 4,245) was similar to those using more standard screw trap methods (approximately 5,400). All species used the faster moving/deeper section of the creek at both SPIs. A backpack PIT tag detector was also developed and used as another remote 'recapture' for additional accuracy in estimating population survival and recapture probability. This unit was used at an approximate efficiency of 24% to survey the creek after the Spring migration. Twenty-five individual fish were re-located. All PIT tag data were used to calculate survival and recapture probabilities using the Cormack-Jolly-Seber population model. Survival for steelhead was high and recapture probability depended greatly on season. Probability of recapture was highest in Spring (29.5%) and relatively low in all other seasons (< 7% in Fall, Winter, and Summer). Wild steelhead PIT tagged in the field and returned to the laboratory had a tag retention rate of 97.6%. A laboratory study was designed to determine the effects of 3-sized PIT tags (12 mm, 20 mm, and 23 mm) on survival and growth of individuals. Survival from surgical implantation of 23 mm PIT tags was > 98% for fish (coho salmon and steelhead). Retention of 23 mm PIT tags was 100% for coho salmon and 89% for steelhead. For both coho and steelhead, growth rates during the first month were affected by tagging, but by the end of 2 months growth effects equalized for all tag sizes. Life history characteristics quantified with SPI techniques are comparable to standard techniques. For example, peaks of Spring migration for steelhead and cutthroat were amazingly similar to those reported from the screw trap. These techniques will enable application of less laborious methods which are more accurate at estimating life history parameters.« less

Evaluation of Fish Movements, Migration Patterns and Populations Abundance with Streamwidth PIT Tag Interrogation Systems, Final Report 2002.

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

Zydlewski, Gayle B.; Casey, Sean

2003-02-01

Two remote Streamwidth PIT tag Interrogation systems (SPIs) were operated continuously for over one year to test the feasibility of these systems for generating movement, migration, survival and smolt production estimates for salmonids. A total of 1,588 juvenile (< 100 mm FL) naturally produced salmonids (7 coho salmon, 482 cutthroat trout, and 1,099 steelhead) were PIT tagged above the upstream-most SPI (9 sites approximately 1 linear km each) in Fall 2001. Age at tagging for wild caught cutthroat and steelhead was 1 year. SPIs were operating before any PIT tagged fish were released in the creek. Over 390,000 detections weremore » recorded from October 2001 to 31 July 2002. Efficiencies were site dependent, but overall detection efficiency for the creek was 97% with 95% confidence intervals of 91-100%. PIT tag detection efficiency ranged from 55-100% depending on the SPI and varied throughout the year with average efficiencies of 73% and 89%. SPI efficiency of PIT tag detection was not completely dependent on electronics noise levels or environmental conditions. Fish from all tagging locations were detected at the SPIs. Steelhead and cutthroat trout were primarily detected moving in the Spring (April-June) coincident with the anticipated smolt migration. Steelhead were also detected moving past SPIs at lower numbers in the Fall and Winter. Travel time between SPIs (downstream movement) was highly dependent on time of year. Travel time in the Spring was significantly faster (34.4 {+-} 7.0 hours) for all species than during any other time of year (763.1 {+-} 267.0 hours). Steelhead and cutthroat migrating in the Spring were the same age as those that did not migrate in the Spring. Peak of steelhead migration recorded at the two SPIs was 5/11 and 5/12 and the peak in the screw trap was recorded on 5/17. Steelhead smolt production estimates using SPIs (3,802 with 95% confidence intervals of 3,440-4,245) was similar to those using more standard screw trap methods (approximately 5,400). All species used the faster moving/deeper section of the creek at both SPIs. A backpack PIT tag detector was also developed and used as another remote ''recapture'' for additional accuracy in estimating population survival and recapture probability. This unit was used at an approximate efficiency of 24% to survey the creek after the Spring migration. Twenty-five individual fish were re-located. All PIT tag data were used to calculate survival and recapture probabilities using the Cormack-Jolly-Seber population model. Survival for steelhead was high and recapture probability depended greatly on season. Probability of recapture was highest in Spring (29.5%) and relatively low in all other seasons (< 7% in Fall, Winter, and Summer). Wild steelhead PIT tagged in the field and returned to the laboratory had a tag retention rate of 97.6%. A laboratory study was designed to determine the effects of 3-sized PIT tags (12 mm, 20 mm, and 23 mm) on survival and growth of individuals. Survival from surgical implantation of 23 mm PIT tags was > 98% for fish (coho salmon and steelhead). Retention of 23 mm PIT tags was 100% for coho salmon and 89% for steelhead. For both coho and steelhead, growth rates during the first month were affected by tagging, but by the end of 2 months growth effects equalized for all tag sizes. Life history characteristics quantified with SPI techniques are comparable to standard techniques. For example, peaks of Spring migration for steelhead and cutthroat were amazingly similar to those reported from the screw trap. These techniques will enable application of less laborious methods which are more accurate at estimating life history parameters.« less

Passage of native riverine fishes through geometrically different sections of a vertical slot fishway on the Moselle River, Germany

USGS Publications Warehouse

Pitsch, Matthias; Mockenhaupt, Bernd; Castro-Santos, Theodore R.

2014-01-01

In order to study effects of different geometric types of pools or change of the flow direction on the passability of fish, sets of PIT antennas were installed inside a modern vertical slot fishway at the mouth of the River Moselle. Fish of 13 abundant species were caught and tagged with PIT tags in 2013 and released in the tailwater of Koblenz. 16% of the tagged fish were detected entering the fishway a short time after release. These individuals provided data on entry and passage rates for different sections of the fishway. Preliminary results show differences in passage time and passage rates between different sections of the fishway.

Multiple sort flow cytometer

DOEpatents

Van den Engh, Ger; Esposito, Richard J.

1996-01-01

A flow cytometer utilizes multiple lasers for excitation and respective fluorescence of identified dyes bonded to specific cells or events to identify and verify multiple events to be sorted from a sheath flow and droplet stream. Once identified, verified and timed in the sheath flow, each event is independently tagged upon separation from the flow by an electrical charge of +60, +120, or +180 volts and passed through oppositely charged deflection plates with ground planes to yield a focused six way deflection of at least six events in a narrow plane.

A new approach to tag design in dolphin telemetry: Computer simulations to minimise deleterious effects

NASA Astrophysics Data System (ADS)

Pavlov, V. V.; Wilson, R. P.; Lucke, K.

2007-02-01

Remote-sensors and transmitters are powerful devices for studying cetaceans at sea. However, despite substantial progress in microelectronics and miniaturisation of systems, dolphin tags are imperfectly designed; additional drag from tags increases swim costs, compromises swimming capacity and manoeuvrability, and leads to extra loads on the animal's tissue. We propose a new approach to tag design, elaborating basic principles and incorporating design stages to minimise device effects by using computer-aided design. Initially, the operational conditions of the device are defined by quantifying the shape, hydrodynamics and range of the natural deformation of the dolphin body at the tag attachment site (such as close to the dorsal fin). Then, parametric models of both of the dorsal fin and a tag are created using the derived data. The link between parameters of the fin and a tag model allows redesign of tag models according to expected changes of fin geometry (difference in fin shape related with species, sex, and age peculiarities, simulation of the bend of the fin during manoeuvres). A final virtual modelling stage uses iterative improvement of a tag model in a computer fluid dynamics (CFD) environment to enhance tag performance. This new method is considered as a suitable tool of tag design before creation of the physical model of a tag and testing with conventional wind/water tunnel technique. Ultimately, tag materials are selected to conform to the conditions identified by the modelling process and thus help create a physical model of a tag, which should minimise its impact on the animal carrier and thus increase the reliability and quality of the data obtained.

Enrichment and Ranking of the YouTube Tag Space and Integration with the Linked Data Cloud

NASA Astrophysics Data System (ADS)

Choudhury, Smitashree; Breslin, John G.; Passant, Alexandre

The increase of personal digital cameras with video functionality and video-enabled camera phones has increased the amount of user-generated videos on the Web. People are spending more and more time viewing online videos as a major source of entertainment and "infotainment". Social websites allow users to assign shared free-form tags to user-generated multimedia resources, thus generating annotations for objects with a minimum amount of effort. Tagging allows communities to organise their multimedia items into browseable sets, but these tags may be poorly chosen and related tags may be omitted. Current techniques to retrieve, integrate and present this media to users are deficient and could do with improvement. In this paper, we describe a framework for semantic enrichment, ranking and integration of web video tags using Semantic Web technologies. Semantic enrichment of folksonomies can bridge the gap between the uncontrolled and flat structures typically found in user-generated content and structures provided by the Semantic Web. The enhancement of tag spaces with semantics has been accomplished through two major tasks: (1) a tag space expansion and ranking step; and (2) through concept matching and integration with the Linked Data cloud. We have explored social, temporal and spatial contexts to enrich and extend the existing tag space. The resulting semantic tag space is modelled via a local graph based on co-occurrence distances for ranking. A ranked tag list is mapped and integrated with the Linked Data cloud through the DBpedia resource repository. Multi-dimensional context filtering for tag expansion means that tag ranking is much easier and it provides less ambiguous tag to concept matching.

Rapid Analysis of Microalgal Triacylglycerols with Direct-Infusion Mass Spectrometry

DOE PAGES

Christensen, Earl; Sudasinghe, Nilusha; Dandamudi, Kodanda Phani Raj; ...

2015-09-01

Cultivation of microalgae has the potential to provide lipid-derived feedstocks for conversion to liquid transportation fuels. Lipid extracts from microalgae are significantly more complex than those of traditional seed oils, and their composition changes significantly throughout the microalgal growth period. With three acyl side chains per molecule, triglycerides (TAGs) are an important fuel precursor, and the distribution of acyl chain composition for TAGs has a significant impact on fuel properties and processing. Therefore, determination of the distribution of microalgal TAG production is needed to assess the value of algal extracts designed for fuel production and to optimize strain, cultivation, andmore » harvesting practices. Methods utilized for TAG speciation commonly involve complicated and time-consuming chromatographic techniques. Here we present a method for TAG speciation and quantification based on direct-infusion mass spectrometry, which provides rapid characterization of TAG profiles without chromatographic separation. Specifically, we utilize Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) to provide a reference library of TAGs for the microalgae Nannochloropsis sp. that provides the basis for high-throughput TAG quantitation by time-of-flight mass spectrometry (TOF MS). In conclusion, we demonstrate the application of this novel approach for lipid characterization with respect to TAG compound distribution, which informs both immediate and future strain and process optimization strategies.« less

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

Christensen, Earl; Sudasinghe, Nilusha; Dandamudi, Kodanda Phani Raj

Cultivation of microalgae has the potential to provide lipid-derived feedstocks for conversion to liquid transportation fuels. Lipid extracts from microalgae are significantly more complex than those of traditional seed oils, and their composition changes significantly throughout the microalgal growth period. With three acyl side chains per molecule, triglycerides (TAGs) are an important fuel precursor, and the distribution of acyl chain composition for TAGs has a significant impact on fuel properties and processing. Therefore, determination of the distribution of microalgal TAG production is needed to assess the value of algal extracts designed for fuel production and to optimize strain, cultivation, andmore » harvesting practices. Methods utilized for TAG speciation commonly involve complicated and time-consuming chromatographic techniques. Here we present a method for TAG speciation and quantification based on direct-infusion mass spectrometry, which provides rapid characterization of TAG profiles without chromatographic separation. Specifically, we utilize Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) to provide a reference library of TAGs for the microalgae Nannochloropsis sp. that provides the basis for high-throughput TAG quantitation by time-of-flight mass spectrometry (TOF MS). In conclusion, we demonstrate the application of this novel approach for lipid characterization with respect to TAG compound distribution, which informs both immediate and future strain and process optimization strategies.« less

Harvesting Intelligence in Multimedia Social Tagging Systems

NASA Astrophysics Data System (ADS)

Giannakidou, Eirini; Kaklidou, Foteini; Chatzilari, Elisavet; Kompatsiaris, Ioannis; Vakali, Athena

As more people adopt tagging practices, social tagging systems tend to form rich knowledge repositories that enable the extraction of patterns reflecting the way content semantics is perceived by the web users. This is of particular importance, especially in the case of multimedia content, since the availability of such content in the web is very high and its efficient retrieval using textual annotations or content-based automatically extracted metadata still remains a challenge. It is argued that complementing multimedia analysis techniques with knowledge drawn from web social annotations may facilitate multimedia content management. This chapter focuses on analyzing tagging patterns and combining them with content feature extraction methods, generating, thus, intelligence from multimedia social tagging systems. Emphasis is placed on using all available "tracks" of knowledge, that is tag co-occurrence together with semantic relations among tags and low-level features of the content. Towards this direction, a survey on the theoretical background and the adopted practices for analysis of multimedia social content are presented. A case study from Flickr illustrates the efficiency of the proposed approach.

Imidazolium tagged acridines: Synthesis, characterization and applications in DNA binding and anti-microbial activities

NASA Astrophysics Data System (ADS)

Raju, Gembali; Vishwanath, S.; Prasad, Archana; Patel, Basant K.; Prabusankar, Ganesan

2016-03-01

New water soluble 4,5-bis imidazolium tagged acridines have been synthesized and structurally characterized by multinuclear NMR and single crystal X-ray diffraction techniques. The DNA binding and anti-microbial activities of these acridine derivatives were investigated by fluorescence and far-UV circular dichroism studies.

A machine learning approach for automated wide-range frequency tagging analysis in embedded neuromonitoring systems.

PubMed

Montagna, Fabio; Buiatti, Marco; Benatti, Simone; Rossi, Davide; Farella, Elisabetta; Benini, Luca

2017-10-01

EEG is a standard non-invasive technique used in neural disease diagnostics and neurosciences. Frequency-tagging is an increasingly popular experimental paradigm that efficiently tests brain function by measuring EEG responses to periodic stimulation. Recently, frequency-tagging paradigms have proven successful with low stimulation frequencies (0.5-6Hz), but the EEG signal is intrinsically noisy in this frequency range, requiring heavy signal processing and significant human intervention for response estimation. This limits the possibility to process the EEG on resource-constrained systems and to design smart EEG based devices for automated diagnostic. We propose an algorithm for artifact removal and automated detection of frequency tagging responses in a wide range of stimulation frequencies, which we test on a visual stimulation protocol. The algorithm is rooted on machine learning based pattern recognition techniques and it is tailored for a new generation parallel ultra low power processing platform (PULP), reaching performance of more that 90% accuracy in the frequency detection even for very low stimulation frequencies (<1Hz) with a power budget of 56mW. Copyright © 2017 Elsevier Inc. All rights reserved.

Modeling the tagged-neutron UXO identification technique using the Geant4 toolkit

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

Zhou Y.; Mitra S.; Zhu X.

2011-10-16

It is proposed to use 14 MeV neutrons tagged by the associated particle neutron time-of-flight technique (APnTOF) to identify the fillers of unexploded ordnances (UXO) by characterizing their carbon, nitrogen and oxygen contents. To facilitate the design and construction of a prototype system, a preliminary simulation model was developed, using the Geant4 toolkit. This work established the toolkit environment for (a) generating tagged neutrons, (b) their transport and interactions within a sample to induce emission and detection of characteristic gamma-rays, and (c) 2D and 3D-image reconstruction of the interrogated object using the neutron and gamma-ray time-of-flight information. Using the modeling,more » this article demonstrates the novelty of the tagged-neutron approach for extracting useful signals with high signal-to-background discrimination of an object-of-interest from that of its environment. Simulations indicated that an UXO filled with the RDX explosive, hexogen (C{sub 3}H{sub 6}O{sub 6}N{sub 6}), can be identified to a depth of 20 cm when buried in soil.« less

Sorting cells of the microalga Chlorococcum littorale with increased triacylglycerol productivity.

PubMed

Cabanelas, Iago Teles Dominguez; van der Zwart, Mathijs; Kleinegris, Dorinde M M; Wijffels, René H; Barbosa, Maria J

2016-01-01

Despite extensive research in the last decades, microalgae are still only economically feasible for high valued markets. Strain improvement is a strategy to increase productivities, hence reducing costs. In this work, we focus on microalgae selection: taking advantage of the natural biological variability of species to select variations based on desired characteristics. We focused on triacylglycerol (TAG), which have applications ranging from biodiesel to high-value omega-3 fatty-acids. Hence, we demonstrated a strategy to sort microalgae cells with increased TAG productivity. 1. We successfully identified sub-populations of cells with increased TAG productivity using Fluorescence assisted cell sorting (FACS). 2. We sequentially sorted cells after repeated cycles of N-starvation, resulting in five sorted populations (S1-S5). 3. The comparison between sorted and original populations showed that S5 had the highest TAG productivity [0.34 against 0.18 g l(-1) day(-1) (original), continuous light]. 4. Original and S5 were compared in lab-scale reactors under simulated summer conditions confirming the increased TAG productivity of S5 (0.4 against 0.2 g l(-1) day(-1)). Biomass composition analyses showed that S5 produced more biomass under N-starvation because of an increase only in TAG content and, flow cytometry showed that our selection removed cells with lower efficiency in producing TAGs. All combined, our results present a successful strategy to improve the TAG productivity of Chlorococcum littorale, without resourcing to genetic manipulation or random mutagenesis. Additionally, the improved TAG productivity of S5 was confirmed under simulated summer conditions, highlighting the industrial potential of S5 for microalgal TAG production.

Evaluation of radioisotope tracer and activation analysis techniques for contamination monitoring in space environment simulation chambers

NASA Technical Reports Server (NTRS)

Smathers, J. B.; Kuykendall, W. E., Jr.; Wright, R. E., Jr.; Marshall, J. R.

1973-01-01

Radioisotope measurement techniques and neutron activation analysis are evaluated for use in identifying and locating contamination sources in space environment simulation chambers. The alpha range method allows the determination of total contaminant concentration in vapor state and condensate state. A Cf-252 neutron activation analysis system for detecting oils and greases tagged with stable elements is described. While neutron activation analysis of tagged contaminants offers specificity, an on-site system is extremely costly to implement and provides only marginal detection sensitivity under even the most favorable conditions.

Fluorescent protein tagging of endogenous protein in brain neurons using CRISPR/Cas9-mediated knock-in and in utero electroporation techniques

PubMed Central

Uemura, Takeshi; Mori, Takuma; Kurihara, Taiga; Kawase, Shiori; Koike, Rie; Satoga, Michiru; Cao, Xueshan; Li, Xue; Yanagawa, Toru; Sakurai, Takayuki; Shindo, Takayuki; Tabuchi, Katsuhiko

2016-01-01

Genome editing is a powerful technique for studying gene functions. CRISPR/Cas9-mediated gene knock-in has recently been applied to various cells and organisms. Here, we successfully knocked in an EGFP coding sequence at the site immediately after the first ATG codon of the β-actin gene in neurons in the brain by the combined use of the CRISPR/Cas9 system and in utero electroporation technique, resulting in the expression of the EGFP-tagged β-actin protein in cortical layer 2/3 pyramidal neurons. We detected EGFP fluorescence signals in the soma and neurites of EGFP knock-in neurons. These signals were particularly abundant in the head of dendritic spines, corresponding to the localization of the endogenous β-actin protein. EGFP knock-in neurons showed no detectable changes in spine density and basic electrophysiological properties. In contrast, exogenously overexpressed EGFP-β-actin showed increased spine density and EPSC frequency, and changed resting membrane potential. Thus, our technique provides a potential tool to elucidate the localization of various endogenous proteins in neurons by epitope tagging without altering neuronal and synaptic functions. This technique can be also useful for introducing a specific mutation into genes to study the function of proteins and genomic elements in brain neurons. PMID:27782168

Fluorescent protein tagging of endogenous protein in brain neurons using CRISPR/Cas9-mediated knock-in and in utero electroporation techniques.

PubMed

Uemura, Takeshi; Mori, Takuma; Kurihara, Taiga; Kawase, Shiori; Koike, Rie; Satoga, Michiru; Cao, Xueshan; Li, Xue; Yanagawa, Toru; Sakurai, Takayuki; Shindo, Takayuki; Tabuchi, Katsuhiko

2016-10-26

Genome editing is a powerful technique for studying gene functions. CRISPR/Cas9-mediated gene knock-in has recently been applied to various cells and organisms. Here, we successfully knocked in an EGFP coding sequence at the site immediately after the first ATG codon of the β-actin gene in neurons in the brain by the combined use of the CRISPR/Cas9 system and in utero electroporation technique, resulting in the expression of the EGFP-tagged β-actin protein in cortical layer 2/3 pyramidal neurons. We detected EGFP fluorescence signals in the soma and neurites of EGFP knock-in neurons. These signals were particularly abundant in the head of dendritic spines, corresponding to the localization of the endogenous β-actin protein. EGFP knock-in neurons showed no detectable changes in spine density and basic electrophysiological properties. In contrast, exogenously overexpressed EGFP-β-actin showed increased spine density and EPSC frequency, and changed resting membrane potential. Thus, our technique provides a potential tool to elucidate the localization of various endogenous proteins in neurons by epitope tagging without altering neuronal and synaptic functions. This technique can be also useful for introducing a specific mutation into genes to study the function of proteins and genomic elements in brain neurons.

Complementary studies of lipid membrane dynamics using iSCAT and super-resolved fluorescence correlation spectroscopy

NASA Astrophysics Data System (ADS)

Reina, Francesco; Galiani, Silvia; Shrestha, Dilip; Sezgin, Erdinc; de Wit, Gabrielle; Cole, Daniel; Lagerholm, B. Christoffer; Kukura, Philipp; Eggeling, Christian

2018-06-01

Observation techniques with high spatial and temporal resolution, such as single-particle tracking based on interferometric scattering (iSCAT) microscopy, and fluorescence correlation spectroscopy applied on a super-resolution STED microscope (STED-FCS), have revealed new insights of the molecular organization of membranes. While delivering complementary information, there are still distinct differences between these techniques, most prominently the use of fluorescent dye tagged probes for STED-FCS and a need for larger scattering gold nanoparticle tags for iSCAT. In this work, we have used lipid analogues tagged with a hybrid fluorescent tag–gold nanoparticle construct, to directly compare the results from STED-FCS and iSCAT measurements of phospholipid diffusion on a homogeneous supported lipid bilayer (SLB). These comparative measurements showed that while the mode of diffusion remained free, at least at the spatial (>40 nm) and temporal (50  ⩽  t  ⩽  100 ms) scales probed, the diffussion coefficient was reduced by 20- to 60-fold when tagging with 20 and 40 nm large gold particles as compared to when using dye tagged lipid analogues. These FCS measurements of hybrid fluorescent tag–gold nanoparticle labeled lipids also revealed that commercially supplied streptavidin-coated gold nanoparticles contain large quantities of free streptavidin. Finally, the values of apparent diffusion coefficients obtained by STED-FCS and iSCAT differed by a factor of 2–3 across the techniques, while relative differences in mobility between different species of lipid analogues considered were identical in both approaches. In conclusion, our experiments reveal that large and potentially cross-linking scattering tags introduce a significant slow-down in diffusion on SLBs but no additional bias, and our labeling approach creates a new way of exploiting complementary information from STED-FCS and iSCAT measurements.

Altered Agonist Sensitivity of a Mutant V2 Receptor Suggests a Novel Therapeutic Strategy for Nephrogenic Diabetes Insipidus

PubMed Central

Erdélyi, László Sándor; Balla, András; Patócs, Attila; Tóth, Miklós; Várnai, Péter

2014-01-01

Loss-of-function mutations of the type 2 vasopressin receptor (V2R) in kidney can lead to nephrogenic diabetes insipidus (NDI). We studied a previously described, but uncharacterized, mutation of the V2R (N321K missense mutation) of a patient with NDI. The properties of the mutant receptor were evaluated. We constructed a highly sensitive Epac-based bioluminescence resonance energy transfer biosensor to perform real-time cAMP measurements after agonist stimulation of transiently transfected HEK293 cells with V2Rs. β-Arrestin binding of the activated receptors was examined with luciferase-tagged β-arrestin and mVenus-tagged V2Rs using the bioluminescence resonance energy transfer technique. Cell surface expression levels of hemagglutinin-tagged receptors were determined with flow cytometry using anti-hemagglutinin-Alexa 488 antibodies. Cellular localization examinations were implemented with fluorescent tagged receptors visualized with confocal laser scanning microscopy. The effect of various vasopressin analogs on the type 1 vasopressin receptor (V1R) was tested on mouse arteries by wire myography. The N321K mutant V2R showed normal cell surface expression, but the potency of arginine vasopressin for cAMP generation was low, whereas the clinically used desmopressin was not efficient. The β-arrestin binding and internalization properties of the mutant receptor were also different than those for the wild type. The function of the mutant receptor can be rescued with administration of the V2R agonist Val4-desmopressin, which had no detectable side effects on V1R in the effective cAMP generating concentrations. Based on these findings we propose a therapeutic strategy for patients with NDI carrying the N321K mutation, as our in vivo experiments suggest that Val4-desmopressin could rescue the function of the N321K-V2R without significant side effects on the V1R. PMID:24628417

Altered agonist sensitivity of a mutant v2 receptor suggests a novel therapeutic strategy for nephrogenic diabetes insipidus.

PubMed

Erdélyi, László Sándor; Balla, András; Patócs, Attila; Tóth, Miklós; Várnai, Péter; Hunyady, László

2014-05-01

Loss-of-function mutations of the type 2 vasopressin receptor (V2R) in kidney can lead to nephrogenic diabetes insipidus (NDI). We studied a previously described, but uncharacterized, mutation of the V2R (N321K missense mutation) of a patient with NDI. The properties of the mutant receptor were evaluated. We constructed a highly sensitive Epac-based bioluminescence resonance energy transfer biosensor to perform real-time cAMP measurements after agonist stimulation of transiently transfected HEK293 cells with V2Rs. β-Arrestin binding of the activated receptors was examined with luciferase-tagged β-arrestin and mVenus-tagged V2Rs using the bioluminescence resonance energy transfer technique. Cell surface expression levels of hemagglutinin-tagged receptors were determined with flow cytometry using anti-hemagglutinin-Alexa 488 antibodies. Cellular localization examinations were implemented with fluorescent tagged receptors visualized with confocal laser scanning microscopy. The effect of various vasopressin analogs on the type 1 vasopressin receptor (V1R) was tested on mouse arteries by wire myography. The N321K mutant V2R showed normal cell surface expression, but the potency of arginine vasopressin for cAMP generation was low, whereas the clinically used desmopressin was not efficient. The β-arrestin binding and internalization properties of the mutant receptor were also different than those for the wild type. The function of the mutant receptor can be rescued with administration of the V2R agonist Val(4)-desmopressin, which had no detectable side effects on V1R in the effective cAMP generating concentrations. Based on these findings we propose a therapeutic strategy for patients with NDI carrying the N321K mutation, as our in vivo experiments suggest that Val(4)-desmopressin could rescue the function of the N321K-V2R without significant side effects on the V1R.

Affinity Purification of Proteins in Tag-Free Form: Split Intein-Mediated Ultrarapid Purification (SIRP).

PubMed

Guan, Dongli; Chen, Zhilei

2017-01-01

Proteins purified using affinity-based chromatography often exploit a recombinant affinity tag. Existing methods for the removal of the extraneous tag, needed for many applications, suffer from poor efficiency and/or high cost. Here we describe a simple, efficient, and potentially low-cost approach-split intein-mediated ultrarapid purification (SIRP)-for both the purification of the desired tagged protein from Escherichia coli lysate and removal of the tag in less than 1 h. The N- and C-fragment of a self-cleaving variant of a naturally split DnaE intein from Nostoc punctiforme are genetically fused to the N-terminus of an affinity tag and a protein of interest (POI), respectively. The N-intein/affinity tag is used to functionalize an affinity resin. The high affinity between the N- and C-fragment of DnaE intein enables the POI to be purified from the lysate via affinity to the resin, and the intein-mediated C-terminal cleavage reaction causes tagless POI to be released into the flow-through. The intein cleavage reaction is strongly inhibited by divalent ions (e.g., Zn 2+ ) under non-reducing conditions and is significantly enhanced by reducing conditions. The POI is cleaved efficiently regardless of the identity of the N-terminal amino acid except in the cases of threonine and proline, and the N-intein-functionalized affinity resin can be regenerated for multiple cycles of use.

The Automatic Integration of Folksonomies with Taxonomies Using Non-axiomatic Logic

NASA Astrophysics Data System (ADS)

Geldart, Joe; Cummins, Stephen

Cooperative tagging systems such as folksonomies are powerful tools when used to annotate information resources. The inherent power of folksonomies is in their ability to allow casual users to easily contribute ad hoc, yet meaningful, resource metadata without any specialist training. Older folksonomies have begun to degrade due to the lack of internal structure and from the use of many low quality tags. This chapter describes a remedy for some of the problems associated with folksonomies. We introduce a method of automatic integration and inference of the relationships between tags and resources in a folksonomy using non-axiomatic logic. We test this method on the CiteULike corpus of tags by comparing precision and recall between it and standard keyword search. Our results show that non-axiomatic reasoning is a promising technique for integrating tagging systems with more structured knowledge representations.

The Field Use Of The EURITRACK Tagged Neutron Inspection System

NASA Astrophysics Data System (ADS)

Viesti, G.; Fabris, D.; Lunardon, M.; Moretto, S.; Nebbia, G.; Pesente, S.

2011-06-01

The EURITRACK portal makes use of the tagged neutron technique to inspect suspect regions of cargo containers identified in preliminary x-ray inspections. The portal has been in operation at the Rijeka seaport during the last three years. The performances of sub-systems, with particular interest on the sealed tube neutron generator, is presented.

Preliminary evaluation of the behavior and movements of adult spring Chinook salmon in the Chehalis River, southwestern Washington, 2014

USGS Publications Warehouse

Liedtke, Theresa L.; Hurst, William R.; Tomka, Ryan G.; Kock, Tobias J.; Zimmerman, Mara S.

2017-01-30

Recent interest in flood control and restoration strategies in the Chehalis River Basin has increased the need to understand the current status and ecology of spring Chinook salmon (Oncorhynchus tshawytscha). Spring Chinook salmon have the longest exposure of all adult Chinook salmon life histories to the low-flow and high water temperature conditions that typically occur during summer. About 100 adult spring Chinook salmon were found dead in the Chehalis River in July and August 2009. Adult Chinook salmon are known to hold in cool-water refugia during warm summer months, but the extent to which spring Chinook salmon might use thermal refugia in the Chehalis River is unknown. A preliminary evaluation of the movements and temperature exposures of adult spring Chinook salmon following their return to the Chehalis River was conducted using radiotelemetry and transmitters equipped with temperature sensors. A total of 12 spring Chinook salmon were captured, radio-tagged, and released in the main-stem Chehalis River between May and late June 2014. Tagged fish were monitored from freshwater entry through the spawning period using a combination of fixedsite monitoring locations and mobile tracking.Water temperature and flow conditions in the main-stem Chehalis River during 2014 were atypical compared to historical averages. Mean monthly water temperatures between March and August 2014 were higher than any decade since 1960 and mean monthly discharge was 90–206 percent of the discharge in previous years. Overall, 92 percent of the tagged fish were detected, with a mean of 102 d in the detection history of tagged fish. Seven tagged fish (58 percent) moved upstream, either shortly after release (5–8 d, 57 percent), or within about a month (34–35 d, 29 percent). One fish (14 percent) remained near the release location for 98 d before moving upstream. The final fates for the seven fish that moved upstream following release included six fish that were assigned a fate of spawner and one fish with an unknown fate. Tagged fish showed limited movements during the peak water temperatures in July and August, and were not frequently detected at sites where water temperatures exceeded 21 °C. The mouths of the Skookumchuck and Newaukum Rivers were commonly used by tagged fish for extended periods during peak water temperatures and study fish with a fate of spawner were last detected in these tributaries.This pilot study represents a substantial contribution to the understanding of spring Chinook salmon in the Chehalis River Basin, and provides information for the design and execution of future evaluations. The water temperatures and flow conditions during the 2014 study period were not typical of the historical conditions in the basin and the numbers of tagged fish monitored was relatively low, so results should be interpreted with those cautions in mind.

Sequential ordering among multicolor fluorophores for protein labeling facility via aggregation-elimination based β-lactam probes.

PubMed

Sadhu, Kalyan K; Mizukami, Shin; Watanabe, Shuji; Kikuchi, Kazuya

2011-05-01

Development of protein labeling techniques with small molecules is enthralling because this method brings promises for triumph over the limitations of fluorescent proteins in live cell imaging. This technology deals with the functionalization of proteins with small molecules and is anticipated to facilitate the expansion of various protein assay methods. A new straightforward aggregation and elimination-based technique for a protein labeling system has been developed with a versatile emissive range of fluorophores. These fluorophores have been applied to show their efficiency for protein labeling by exploiting the same basic principle. A genetically modified version of class A type β-lactamase has been used as the tag protein (BL-tag). The strength of the aggregation interaction between a fluorophore and a quencher plays a governing role in the elimination step of the quencher from the probes, which ultimately controls the swiftness of the protein labeling strategy. Modulation in the elimination process can be accomplished by the variation in the nature of the fluorophore. This diversity facilitates the study of the competitive binding order among the synthesized probes toward the BL-tag labeling method. An aggregation and elimination-based BL-tag technique has been explored to develop an order of color labeling from the equimolar mixture of the labeling probe in solutions. The qualitative and quantitative determination of ordering within the probes toward labeling studies has been executed through SDS-PAGE and time-dependent fluorescence intensity enhancement measurements, respectively. The desirable multiple-wavelength fluorescence labeling probes for the BL-tag technology have been developed and demonstrate broad applicability of this labeling technology to live cell imaging with coumarin and fluorescein derivatives by using confocal microscopy.

Comparison of NxTAG Respiratory Pathogen Panel and Anyplex II RV16 Tests for Multiplex Detection of Respiratory Pathogens in Hospitalized Children

PubMed Central

Brotons, Pedro; Henares, Desiree; Latorre, Irene; Cepillo, Antonio; Launes, Cristian

2016-01-01

Multiplex molecular techniques can detect a diversity of respiratory viruses and bacteria that cause childhood acute respiratory infection rapidly and conveniently. However, currently available techniques show high variation in performance. We sought to compare the diagnostic accuracy of the novel multiplex NxTAG respiratory pathogen panel (RPP) RUO test versus a routine multiplex Anyplex II RV16 assay in respiratory specimens collected from children <18 years of age hospitalized with nonspecific symptoms of acute lower respiratory infection. Parallel testing was performed on nasopharyngeal aspirates prospectively collected at referral Children's Hospital Sant Joan de Déu (Barcelona, Spain) between June and November 2015. Agreement values between the two tests and kappa coefficients were assessed. Bidirectional sequencing was performed for the resolution of discordant results. A total of 319 samples were analyzed by both techniques. A total of 268 (84.0%) of them yielded concordant results. Positive percent agreement values ranged from 83.3 to 100%, while the negative percent agreement was more than 99% for all targets except for enterovirus/rhinovirus (EV/RV; 94.4%). Kappa coefficients ranged from 0.83 to 1.00. Discrepancy analysis confirmed 66.0% of NxTAG RPP RUO results. A total of 260 viruses were detected, with EV/RV (n = 105, 40.4%) being the most prevalent target. Viral coinfections were found in 44 (14.2%) samples. In addition, NxTAG RPP RUO detected single bacterial and mixed viral-bacterial infections in seven samples. NxTAG RPP RUO showed high positive and negative agreement with Anyplex II RV16 for main viruses that cause acute respiratory infections in children, coupled with an additional capability to detect some respiratory bacteria. PMID:27629904

Characterizing pulmonary blood flow distribution measured using arterial spin labeling.

PubMed

Henderson, A Cortney; Prisk, G Kim; Levin, David L; Hopkins, Susan R; Buxton, Richard B

2009-12-01

The arterial spin labeling (ASL) method provides images in which, ideally, the signal intensity of each image voxel is proportional to the local perfusion. For studies of pulmonary perfusion, the relative dispersion (RD, standard deviation/mean) of the ASL signal across a lung section is used as a reliable measure of flow heterogeneity. However, the RD of the ASL signals within the lung may systematically differ from the true RD of perfusion because the ASL image also includes signals from larger vessels, which can reflect the blood volume rather than blood flow if the vessels are filled with tagged blood during the imaging time. Theoretical studies suggest that the pulmonary vasculature exhibits a lognormal distribution for blood flow and thus an appropriate measure of heterogeneity is the geometric standard deviation (GSD). To test whether the ASL signal exhibits a lognormal distribution for pulmonary blood flow, determine whether larger vessels play an important role in the distribution, and extract physiologically relevant measures of heterogeneity from the ASL signal, we quantified the ASL signal before and after an intervention (head-down tilt) in six subjects. The distribution of ASL signal was better characterized by a lognormal distribution than a normal distribution, reducing the mean squared error by 72% (p < 0.005). Head-down tilt significantly reduced the lognormal scale parameter (p = 0.01) but not the shape parameter or GSD. The RD increased post-tilt and remained significantly elevated (by 17%, p < 0.05). Test case results and mathematical simulations suggest that RD is more sensitive than the GSD to ASL signal from tagged blood in larger vessels, a probable explanation of the change in RD without a statistically significant change in GSD. This suggests that the GSD is a useful measure of pulmonary blood flow heterogeneity with the advantage of being less affected by the ASL signal from tagged blood in larger vessels.

A navigation system for the visually impaired an intelligent white cane.

PubMed

Fukasawa, A Jin; Magatani, Kazusihge

2012-01-01

In this paper, we describe about a developed navigation system that supports the independent walking of the visually impaired in the indoor space. Our developed instrument consists of a navigation system and a map information system. These systems are installed on a white cane. Our navigation system can follow a colored navigation line that is set on the floor. In this system, a color sensor installed on the tip of a white cane, this sensor senses a color of navigation line and the system informs the visually impaired that he/she is walking along the navigation line by vibration. This color recognition system is controlled by a one-chip microprocessor. RFID tags and a receiver for these tags are used in the map information system. RFID tags are set on the colored navigation line. An antenna for RFID tags and a tag receiver are also installed on a white cane. The receiver receives the area information as a tag-number and notifies map information to the user by mp3 formatted pre-recorded voice. And now, we developed the direction identification technique. Using this technique, we can detect a user's walking direction. A triaxiality acceleration sensor is used in this system. Three normal subjects who were blindfolded with an eye mask were tested with our developed navigation system. All of them were able to walk along the navigation line perfectly. We think that the performance of the system is good. Therefore, our system will be extremely valuable in supporting the activities of the visually impaired.

MR colonography with fecal tagging: do individual patient characteristics influence image quality?

PubMed

Kinner, Sonja; Kuehle, Christiane A; Langhorst, Jost; Ladd, Susanne C; Nuefer, Michael; Barkhausen, Joerg; Lauenstein, Thomas C

2007-05-01

To evaluate if different patient characteristics influence performance of fecal tagging (a new MR colonography (MRC) technique to label stool to avoid bowel cleansing) and, consecutively, MR image quality. A total of 333 patients (mean age = 61 years) underwent MRC with fecal tagging. Four segments of the large bowel (ascending, transverse, descending, and sigmoid colon) were assessed as for the presence of nontagged stool particles, which can impede an assessment of the colonic wall. Ratings were correlated with patients' characteristics including patient age (<55 vs. > or =55 years), body mass index (BMI) (<25 vs. > or =25), gender, and acceptance levels for fecal tagging. Statistical analysis was performed using a Mann-Whitney U-test. A total of 1332 colonic segments were evaluated. Among them, 327 segments (25%) did not contain any visible stool particles. Considerably reduced image quality was found in 61 segments (5%). Best image quality was found in the sigmoid colon (mean value = 1.9), while image quality of the ascending colon turned out to be worst (mean value = 2.6). Fecal tagging effectiveness showed a reverse correlation with patient age. However, all other characteristics did not have a statistically significant influence on fecal tagging outcome. MRC in conjunction with barium-based fecal tagging led to diagnostic image quality in 95% of all colonic segments. Since tagging results were significantly decreased in patients > or =55 years, tagging protocols should to be modified in this group, i.e., by increasing the time interval of tagging administration. (c) 2007 Wiley-Liss, Inc.

Evaluation of harmonic direction-finding systems for detecting locomotor activity

USGS Publications Warehouse

Boyarski, V.L.; Rodda, G.H.; Savidge, J.A.

2007-01-01

We conducted a physical simulation experiment to test the efficacy of harmonic direction finding for remotely detecting locomotor activity in animals. The ability to remotely detect movement helps to avoid disturbing natural movement behavior. Remote detection implies that the observer can sense only a change in signal bearing. In our simulated movements, small changes in bearing (<5.7??) were routinely undetectable. Detectability improved progressively with the size of the simulated animal movement. The average (??SD) of reflector tag movements correctly detected for 5 observers was 93.9 ?? 12.8% when the tag was moved ???11.5??; most observers correctly detected tag movements ???20.1??. Given our data, one can assess whether the technique will be effective for detecting movements at an observation distance appropriate for the study organism. We recommend that both habitat and behavior of the organism be taken into consideration when contemplating use of this technique for detecting locomotion.

Monte Carlo analysis of tagged neutron beams for cargo container inspection.

PubMed

Pesente, S; Lunardon, M; Nebbia, G; Viesti, G; Sudac, D; Valkovic, V

2007-12-01

Fast neutrons produced via D+T reactions and tagged by the associated particle technique have been recently proposed to inspect cargo containers. The general characteristics of this technique are studied with Monte Carlo simulations by determining the properties of the tagged neutron beams as a function of the relevant design parameters (energy and size of the deuteron beam, geometry of the charged particle detector). Results from simulations, validated by experiments, show that the broadening of the correlation between the alpha-particle and the neutron, induced by kinematical as well as geometrical (beam and detector size) effects, is important and limits the dimension of the minimum voxel to be inspected. Moreover, the effect of the container filling is explored. The material filling produces a sizeable loss of correlation between alpha-particles and neutrons due to scattering and absorption. Conditions in inspecting cargo containers are discussed.

Bright monomeric photoactivatable red fluorescent protein for two-color super-resolution sptPALM of live cells.

PubMed

Subach, Fedor V; Patterson, George H; Renz, Malte; Lippincott-Schwartz, Jennifer; Verkhusha, Vladislav V

2010-05-12

Rapidly emerging techniques of super-resolution single-molecule microscopy of living cells rely on the continued development of genetically encoded photoactivatable fluorescent proteins. On the basis of monomeric TagRFP, we have developed a photoactivatable TagRFP protein that is initially dark but becomes red fluorescent after violet light irradiation. Compared to other monomeric dark-to-red photoactivatable proteins including PAmCherry, PATagRFP has substantially higher molecular brightness, better pH stability, substantially less sensitivity to blue light, and better photostability in both ensemble and single-molecule modes. Spectroscopic analysis suggests that PATagRFP photoactivation is a two-step photochemical process involving sequential one-photon absorbance by two distinct chromophore forms. True monomeric behavior, absence of green fluorescence, and single-molecule performance in live cells make PATagRFP an excellent protein tag for two-color imaging techniques, including conventional diffraction-limited photoactivation microscopy, super-resolution photoactivated localization microscopy (PALM), and single particle tracking PALM (sptPALM) of living cells. Two-color sptPALM imaging was demonstrated using several PATagRFP tagged transmembrane proteins together with PAGFP-tagged clathrin light chain. Analysis of the resulting sptPALM images revealed that single-molecule transmembrane proteins, which are internalized into a cell via endocytosis, colocalize in space and time with plasma membrane domains enriched in clathrin light-chain molecules.

Frequency-Modulated Continuous Flow Analysis Electrospray Ionization Mass Spectrometry (FM-CFA-ESI-MS) for Sample Multiplexing.

PubMed

Filla, Robert T; Schrell, Adrian M; Coulton, John B; Edwards, James L; Roper, Michael G

2018-02-20

A method for multiplexed sample analysis by mass spectrometry without the need for chemical tagging is presented. In this new method, each sample is pulsed at unique frequencies, mixed, and delivered to the mass spectrometer while maintaining a constant total flow rate. Reconstructed ion currents are then a time-dependent signal consisting of the sum of the ion currents from the various samples. Spectral deconvolution of each reconstructed ion current reveals the identity of each sample, encoded by its unique frequency, and its concentration encoded by the peak height in the frequency domain. This technique is different from other approaches that have been described, which have used modulation techniques to increase the signal-to-noise ratio of a single sample. As proof of concept of this new method, two samples containing up to 9 analytes were multiplexed. The linear dynamic range of the calibration curve was increased with extended acquisition times of the experiment and longer oscillation periods of the samples. Because of the combination of the samples, salt had little effect on the ability of this method to achieve relative quantitation. Continued development of this method is expected to allow for increased numbers of samples that can be multiplexed.

Multiple sort flow cytometer

DOEpatents

Engh, G. van den; Esposito, R.J.

1996-01-09

A flow cytometer utilizes multiple lasers for excitation and respective fluorescence of identified dyes bonded to specific cells or events to identify and verify multiple events to be sorted from a sheath flow and droplet stream. Once identified, verified and timed in the sheath flow, each event is independently tagged upon separation from the flow by an electrical charge of +60, +120, or +180 volts and passed through oppositely charged deflection plates with ground planes to yield a focused six way deflection of at least six events in a narrow plane. 8 figs.

A nu-space for image correlation spectroscopy: characterization and application to measure protein transport in live cells

NASA Astrophysics Data System (ADS)

Potvin-Trottier, Laurent; Chen, Lingfeng; Horwitz, Alan Rick; Wiseman, Paul W.

2013-08-01

We introduce a new generalized theoretical framework for image correlation spectroscopy (ICS). Using this framework, we extend the ICS method in time-frequency (ν, nu) space to map molecular flow of fluorescently tagged proteins in individual living cells. Even in the presence of a dominant immobile population of fluorescent molecules, nu-space ICS (nICS) provides an unbiased velocity measurement, as well as the diffusion coefficient of the flow, without requiring filtering. We also develop and characterize a tunable frequency-filter for spatio-temporal ICS (STICS) that allows quantification of the density, the diffusion coefficient and the velocity of biased diffusion. We show that the techniques are accurate over a wide range of parameter space in computer simulation. We then characterize the retrograde flow of adhesion proteins (α6- and αLβ2-GFP integrins and mCherry-paxillin) in CHO.B2 cells plated on laminin and intercellular adhesion molecule 1 (ICAM-1) ligands respectively. STICS with a tunable frequency filter, in conjunction with nICS, measures two new transport parameters, the density and transport bias coefficient (a measure of the diffusive character of a flow/biased diffusion), showing that molecular flow in this cell system has a significant diffusive component. Our results suggest that the integrin-ligand interaction, along with the internal myosin-motor generated force, varies for different integrin-ligand pairs, consistent with previous results.

Hypersonic Boundary Layer Measurements with Variable Blowing Rates Using Molecular Tagging Velocimetry

NASA Technical Reports Server (NTRS)

Bathel, Brett F.; Danehy, Paul M.; Johansen, Craig T.; Jones, Stephen B.; Goyne, Christopher P.

2012-01-01

Measurements of mean and instantaneous streamwise velocity profiles in a hypersonic boundary layer with variable rates of mass injection (blowing) of nitrogen dioxide (NO2) were obtained over a 10-degree half-angle wedge model. The NO2 was seeded into the flow from a slot located 29.4 mm downstream of the sharp leading edge. The top surface of the wedge was oriented at a 20 degree angle in the Mach 10 flow, yielding an edge Mach number of approximately 4.2. The streamwise velocity profiles and streamwise fluctuating velocity component profiles were obtained using a three-laser NO2->NO photolysis molecular tagging velocimetry method. Observed trends in the mean streamwise velocity profiles and profiles of the fluctuating component of streamwise velocity as functions of the blowing rate are described. An effort is made to distinguish between the effect of blowing rate and wall temperature on the measured profiles. An analysis of the mean velocity profiles for a constant blowing rate is presented to determine the uncertainty in the measurement for different probe laser delay settings. Measurements of streamwise velocity were made to within approximately 120 gm of the model surface. The streamwise spatial resolution in this experiment ranged from 0.6 mm to 2.6 mm. An improvement in the spatial precision of the measurement technique has been made, with spatial uncertainties reduced by about a factor of 2 compared to previous measurements. For the quiescent flow calibration measurements presented, uncertainties as low as 2 m/s are obtained at 95% confidence for long delay times (25 gs). For the velocity measurements obtained with the wind tunnel operating, average single-shot uncertainties of less than 44 m/s are obtained at 95% confidence with a probe laser delay setting of 1 gs. The measurements were performed in the 31-inch Mach 10 Air Tunnel at the NASA Langley Research Center.

Performance studies of D-meson tagged jets in pp collisions at \\sqrt{s}=7\\,{TeV} with ALICE

NASA Astrophysics Data System (ADS)

Aiola, Salvatore; ALICE Collaboration

2017-04-01

We present the current status of the measurement of jets that contain a D meson (D-tagged jets) with the ALICE detector. D0-meson candidates, identified via their hadronic decay into a Kπ pair, were combined with the other charged tracks reconstructed with the central tracking system, using the anti-kT jet-finding algorithm. The yield of D-tagged jets was extracted through an invariant mass analysis of the D-meson candidates. A Monte Carlo simulation was used to determine the detector performance and validate the signal extraction techniques.

Report A: Fish distribution and population dynamics in Rock Creek, Klickitat County, Washington

USGS Publications Warehouse

Allen, Brady; Munz, Carrie S.; Harvey, Elaine

2013-01-01

The U.S. Geological Survey collaborated with the Yakama Nation starting in fall of 2009 to study the fish populations in Rock Creek, a Washington State tributary of the Columbia River 21 kilometers upstream of John Day Dam. Prior to this study, very little was known about the ESA-listed (threatened) Mid-Columbia River steelhead (Oncorhynchus mykiss) population in this arid watershed with intermittent stream flow. The objectives of the study were to quantify fish habitat, document fish distribution, abundance, and movement, and identify areas of high salmonid productivity. To accomplish these objectives, we electrofished in the spring and fall, documenting the distribution and relative abundance of all fish species to evaluate the influence of biotic factors on salmonid productivity and survival. We surveyed the distribution of perennial pools and established a network of automated temperature recording devices from river kilometer (rkm) 2 to 23 in Rock Creek and rkm 0 to 8 in Squaw Creek, a major tributary entering Rock Creek at rkm 13, to better understand the abiotic factors influencing the salmonid populations. Salmonid abundance estimates were conducted using a mark-recapture method in a systematic subsample of the perennial pools. The proportion and timing of salmonids migrating from these pools were assessed by building, installing, and operating two passive integrated transponder (PIT) tag interrogation systems at rkm 5 and at the confluence with Squaw Creek (rkm 13). From fall 2009 to fall 2012, we PIT-tagged 3,088 O. mykiss and 151 coho salmon (O. kisutch) during electrofishing efforts. In the lowest flow periods of 2010 to 2012, we found that an average of 36% of the surveyed streambed length was dry, and 17% remained as perennial pools. The maximum temperature recorded in those pools was 24.4°C, but most pools had a maximum temperature that was less than 21°C. O. mykiss were present in most pools, and non-native fish species, such as smallmouth bass (Micropterus dolomieu), were typically found downstream of rkm 5. Coho salmon were present in nearly every pool that was sampled in 2011, but were rare in 2009, 2010, and 2012. About 27% of the PIT-tagged O. mykiss and 38% of the PIT-tagged coho were detected outmigrating to the Columbia River. Of those fish, 92% (n=695) were detected leaving Rock Creek as smolts in April and May. As of November 2013, 9 O. mykiss and 4 coho that we tagged in Rock Creek as juveniles have returned as adults to Bonneville Dam. Also, an additional 34 PIT-tagged adult steelhead, and 6 PIT-tagged coho that were tagged by other groups have been detected in Rock Creek, of which, 22 were of known origin (tagged as juveniles). Of these, 85% were tagged or released in the Snake River. The PIT-tag interrogation systems will be operated for several more years to allow time for the fish tagged as juveniles to return as adults and complete their life cycles. The Yakama Nation will use the information collected from this study to prioritize and gauge the effectiveness of ongoing and future restoration actions.

Tracking intracellular uptake and localisation of alkyne tagged fatty acids using Raman spectroscopy

NASA Astrophysics Data System (ADS)

Jamieson, Lauren E.; Greaves, Jennifer; McLellan, Jayde A.; Munro, Kevin R.; Tomkinson, Nicholas C. O.; Chamberlain, Luke H.; Faulds, Karen; Graham, Duncan

2018-05-01

Intracellular uptake, distribution and metabolism of lipids are tightly regulated characteristics in healthy cells. An analytical technique capable of understanding these characteristics with a high level of species specificity in a minimally invasive manner is highly desirable in order to understand better how these become disrupted during disease. In this study, the uptake and distribution of three different alkyne tagged fatty acids in single cells were monitored and compared, highlighting the ability of Raman spectroscopy combined with alkyne tags for better understanding of the fine details with regard to uptake, distribution and metabolism of very chemically specific lipid species. This indicates the promise of using Raman spectroscopy directly with alkyne tagged lipids for cellular studies as opposed to subsequently clicking of a fluorophore onto the alkyne for fluorescence imaging.

Study of Valproic Acid-Enhanced Hepatocyte Steatosis

PubMed Central

Chang, Renin; Chou, Mei-Chia; Hung, Li-Ying; Wang, Mu-En; Hsu, Meng-Chieh; Chiu, Chih-Hsien

2016-01-01

Valproic acid (VPA) is one of the most widely used antiepilepsy drugs. However, several side effects, including weight gain and fatty liver, have been reported in patients following VPA treatment. In this study, we explored the molecular mechanisms of VPA-induced hepatic steatosis using FL83B cell line-based in vitro model. Using fluorescent lipid staining technique, we found that VPA enhanced oleic acid- (OLA-) induced lipid accumulation in a dose-dependent manner in hepatocytes; this may be due to upregulated lipid uptake, triacylglycerol (TAG) synthesis, and lipid droplet formation. Real-time PCR results showed that, following VPA treatment, the expression levels of genes encoding cluster of differentiation 36 (Cd36), low-density lipoprotein receptor-related protein 1 (Lrp1), diacylglycerol acyltransferase 2 (Dgat2), and perilipin 2 (Plin2) were increased, that of carnitine palmitoyltransferase I a (Cpt1a) was not affected, and those of acetyl-Co A carboxylase α (Acca) and fatty acid synthase (Fasn) were decreased. Furthermore, using immunofluorescence staining and flow cytometry analyses, we found that VPA also induced peroxisome proliferator-activated receptor γ (PPARγ) nuclear translocation and increased levels of cell-surface CD36. Based on these results, we propose that VPA may enhance OLA-induced hepatocyte steatosis through the upregulation of PPARγ- and CD36-dependent lipid uptake, TAG synthesis, and lipid droplet formation. PMID:27034954

Fabrication of hierarchical hybrid structures using bio-enabled layer-by-layer self-assembly.

PubMed

Hnilova, Marketa; Karaca, Banu Taktak; Park, James; Jia, Carol; Wilson, Brandon R; Sarikaya, Mehmet; Tamerler, Candan

2012-05-01

Development of versatile and flexible assembly systems for fabrication of functional hybrid nanomaterials with well-defined hierarchical and spatial organization is of a significant importance in practical nanobiotechnology applications. Here we demonstrate a bio-enabled self-assembly technique for fabrication of multi-layered protein and nanometallic assemblies utilizing a modular gold-binding (AuBP1) fusion tag. To accomplish the bottom-up assembly we first genetically fused the AuBP1 peptide sequence to the C'-terminus of maltose-binding protein (MBP) using two different linkers to produce MBP-AuBP1 hetero-functional constructs. Using various spectroscopic techniques, surface plasmon resonance (SPR) and localized surface plasmon resonance (LSPR), we verified the exceptional binding and self-assembly characteristics of AuBP1 peptide. The AuBP1 peptide tag can direct the organization of recombinant MBP protein on various gold surfaces through an efficient control of the organic-inorganic interface at the molecular level. Furthermore using a combination of soft-lithography, self-assembly techniques and advanced AuBP1 peptide tag technology, we produced spatially and hierarchically controlled protein multi-layered assemblies on gold nanoparticle arrays with high molecular packing density and pattering efficiency in simple, reproducible steps. This model system offers layer-by-layer assembly capability based on specific AuBP1 peptide tag and constitutes novel biological routes for biofabrication of various protein arrays, plasmon-active nanometallic assemblies and devices with controlled organization, packing density and architecture. Copyright © 2011 Wiley Periodicals, Inc.

The use of a 'bleach-etch-seal' deproteinization technique on MIH affected enamel.

PubMed

Gandhi, Shan; Crawford, Peter; Shellis, Peter

2012-11-01

To ascertain whether deproteinization pretreatment of molar-incisor hypomineralization (MIH) enamel affects resin sealant infiltration. Thirty one extracted MIH teeth were divided into three sections and randomly allocated into the Control (etch and FS), Treatment 1 (5% NaOCl, etched and fissure sealed), and Treatment 2 (5% NaOCl and fissure sealed with no etch) groups. Two hundred seventy nine sealant tag/enamel grade observations were recorded by scanning electron microscopy. Control and Treatment 1 were similar in their outcomes, and Treatment 2 was markedly different. There was no statistical evidence to suggest that there was any difference between Treatment 1 and the Control Treatment (95% CI, 0.52, 1.51; P = 0.6). There was a marked difference between Treatment 2 and the Control Treatment (95% CI, 0.07, 0.25; P < 0.001). All treatments also demonstrated a high-predicted probability of obtaining 'poor' sealant tags (Control = 47%, Treatment 1 = 49%, and Treatment 2 = 40%). The findings suggest that there was no significant difference in the tag quality between the conventional technique (Control) and the 'bleach-etch-seal' technique (Treatment 1). There was no benefit in pre-treating with NaOCl alone (without etch) before sealing. This research also showed that there was a high-predicted probability of obtaining 'poor' sealant tags in MIH enamel, regardless of which of the three treatments was used. © 2012 The Authors. International Journal of Paediatric Dentistry © 2012 BSPD, IAPD and Blackwell Publishing Ltd.

Additional Value of Transluminal Attenuation Gradient in CT Angiography to Predict Hemodynamic Significance of Coronary Artery Stenosis

PubMed Central

Stuijfzand, Wynand J.; Danad, Ibrahim; Raijmakers, Pieter G.; Marcu, C. Bogdan; Heymans, Martijn W.; van Kuijk, Cornelis C.; van Rossum, Albert C.; Nieman, Koen; Min, James K.; Leipsic, Jonathon; van Royen, Niels; Knaapen, Paul

2015-01-01

OBJECTIVES The current study evaluates the incremental value of transluminal attenuation gradient (TAG), TAG with corrected contrast opacification (CCO), and TAG with exclusion of calcified coronary segments (ExC) over coronary computed tomography angiogram (CTA) alone using fractional flow reserve (FFR) as the gold standard. BACKGROUND TAG is defined as the contrast opacification gradient along the length of a coronary artery on a coronary CTA. Preliminary data suggest that TAG provides additional functional information. Interpretation of TAG is hampered by multiple heartbeat acquisition algorithms and coronary calcifications. Two correction models have been proposed based on either dephasing of contrast delivery by relating coronary density to corresponding descending aortic opacification (TAG-CCO) or excluding calcified coronary segments (TAG-ExC). METHODS Eighty-five patients with intermediate probability of coronary artery disease were prospectively included. All patients underwent step-and-shoot 256-slice coronary CTA. TAG, TAG-CCO, and TAG-ExC analyses were performed followed by invasive coronary angiography in conjunction with FFR measurements of all major coronary branches. RESULTS Thirty-four patients (40%) were diagnosed with hemodynamically-significant coronary artery disease (i.e., FFR ≤0.80). On a per-vessel basis (n = 253), 59 lesions (23%) were graded as hemodynamically significant, and the diagnostic accuracy of coronary CTA (diameter stenosis ≥50%) was 95%, 75%, 98%, and 54% for sensitivity, specificity, negative predictive value, and positive predictive value, respectively. TAG and TAG-ExC did not discriminate between vessels with or without hemodynamically significant lesions (−13.5 ± 17.1 HU [Hounsfield units] × 10 mm−1 vs. −11.6 ± 13.3 HU × 10 mm−1, p = 0.36; and 13.1 ± 15.9 HU × 10 mm−1 vs. −11.4 ± 11.7 HU × 10 mm−1, p = 0.77, respectively). TAG-CCO was lower in vessels with a hemodynamically-significant lesion (−0.050 ± 0.051 10 mm−1 vs. −0.036 ± 0.034 10 mm−1, p = 0.03) and TAG-ExC resulted in a slight improvement of the net reclassification index (0.021, p < 0.05). CONCLUSIONS TAG did not provide incremental diagnostic value over 256-slice coronary CTA alone in assessing the hemodynamic consequences of a coronary stenosis. Correction for temporal nonuniformity of contrast delivery or exclusion of calcified coronary segments slightly enhanced the results. PMID:24631509

Comparison of 180-degree and 90-degree needle rotation to reduce wound size in PIT-injected juvenile Chinook salmon

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

Bryson, Amanda J.; Woodley, Christa M.; Karls, Rhonda K.

2013-04-30

Animal telemetry, which requires the implantation of passive transponders or active transmitters, is used to monitor and assess fish stock and conservation to gain an understanding of fish movement and behavior. As new telemetry technologies become available, studies of their effects on species of interest are imperative as is development of implantation techniques. In this study, we investigated the effects of bevel rotation (0-, 90-, 180-degree axis rotation) on wound extent, tag loss, and wound healing rates in juvenile Chinook salmon injected with an 8-gauge needle, which is required for implantation of the novel injectable Juvenile Salmon Acoustic Telemetry Systemsmore » (JSATS) acoustic transmitter or large passive integrated transponder (PIT) tags. Although the injection sites were not closed after injection (e.g., with sutures or glue), there were no mortalities, dropped tags, or indications of fungus, ulceration, and/or redness around the wound. On Day 0 and post-implantation Day 7, the 90-degree bevel rotation produced smaller wound extent than the 180-degree bevel rotation. No axis rotation (0-degrees) resulted in the PIT tag frequently misleading or falling out upon injection. The results of this study indicated the 90-degree bevel rotation was the more efficient technique, produced less wound extent. Given the wound extent compared to size of fish, we recommend researchers should consider a 90-degree rotation over the 180-degree rotation in telemetry studies. Highlights •Three degrees of needle rotation were examined for effects in Chinook salmon. •Mortality, tag loss, wound extent, healing, and infection indicators were measured. •There were no mortalities, tag loss, or indications of infection. •The 90-degree needle rotation through Day 7 produced the smallest wound extent.« less

Elastin-like polypeptides: A strategic fusion partner for biologics.

PubMed

Yeboah, Agnes; Cohen, Rick I; Rabolli, Charles; Yarmush, Martin L; Berthiaume, Francois

2016-08-01

Elastin-like peptides (ELPs) are derivatives of tropoelastin with a unique property that allows them to stay soluble below a certain critical temperature but reversibly form aggregates above that temperature. Since they are derived from tropoelastin, ELPs are biocompatible, non-toxic, and non-immunogenic. The unique properties of ELPs have made them a desirable class of fusion tags used in several biomedical applications including targeted drug delivery and enhancing the half-life of protein drugs. The most significant property of an ELP is that when fused to other proteins, the phase transition property of the ELP is maintained, and the target protein can be purified using the thermally driven property of the ELP. The ELP tag purification process is simple and inexpensive, and involves cycling the protein above and below the transition temperature of the ELP fusion followed by centrifugation to obtain the desired protein, without any need for chromatography. Consequently, using ELPs as a purification tag should be potentially interesting to biopharmaceutical companies who spend a significant percentage of their manufacturing costs on expensive protein purification techniques such as chromatography and filtration. However, ELP tags have not yet been used for commercial protein purification due to some challenges of translating this technique, which has been demonstrated mostly in academic laboratories, to a biotechnology manufacturing environment. The article first reviews the state-of-the-art in protein "ELPylation," and discusses some applications which have benefitted from using ELP as a fusion tag. Then, the article discusses the main advantages of using ELP as a purification tag, and evaluates the remaining hurdles for its implementation in industrial protein production. Biotechnol. Bioeng. 2016;113: 1617-1627. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Fun With Food Webs

ERIC Educational Resources Information Center

Smith, Karl D.

1977-01-01

Explains an upper elementary game of tag that illustrates energy flow in food webs using candy bars as food sources. A follow-up field trip to a river and five language arts projects are also suggested. (CS)

Design and testing of RFID sensor tag fabricated using inkjet-printing and electrodeposition

NASA Astrophysics Data System (ADS)

Chien Dang, Mau; Son Nguyen, Dat; Dung Dang, Thi My; Tedjini, Smail; Fribourg-Blanc, Eric

2014-06-01

The passive RFID tag with an added sensing function is of interest to many applications. In particular, applications where RFID tagging is already considered to be the next step, such as food items, are a specific target. This paper demonstrates a flexible RFID tag sensor fabricated using a low cost technique with an added zero-cost sensing function. It is more specifically applied to the sensing of degradable food, in particular beef meat in our demonstrated example. To reach this, the antenna is designed in such a way to be sensitive to the variation of the dielectric permittivity of the meat over time. The design of the sensing tag as well as its fabrication process are described. The fabrication involves inkjet printing of a silver nanoparticle based ink on a commercial low cost PET film to create a seed layer. It is followed by a copper electrodeposition step on top of the silver pattern to complete the tag to obtain the desired thickness and conductivity of the tag antenna. The results of the electrical tests showed that with the inkjet printing-electrodeposition combination it is possible to produce flexible electrically conductive patterns for practical RFID applications. The tag was then tested in close-to-real-world conditions and it is demonstrated that it can provide a sensing function to detect the consumption limit of the packaged beef.

Identification of triacylglycerol using automated annotation of high resolution multistage mass spectral trees.

PubMed

Wang, Xiupin; Peng, Qingzhi; Li, Peiwu; Zhang, Qi; Ding, Xiaoxia; Zhang, Wen; Zhang, Liangxiao

2016-10-12

High complexity of identification for non-target triacylglycerols (TAGs) is a major challenge in lipidomics analysis. To identify non-target TAGs, a powerful tool named accurate MS(n) spectrometry generating so-called ion trees is used. In this paper, we presented a technique for efficient structural elucidation of TAGs on MS(n) spectral trees produced by LTQ Orbitrap MS(n), which was implemented as an open source software package, or TIT. The TIT software was used to support automatic annotation of non-target TAGs on MS(n) ion trees from a self-built fragment ion database. This database includes 19108 simulate TAG molecules from a random combination of fatty acids and corresponding 500582 self-built multistage fragment ions (MS ≤ 3). Our software can identify TAGs using a "stage-by-stage elimination" strategy. By utilizing the MS(1) accurate mass and referenced RKMD, the TIT software can discriminate unique elemental composition candidates. The regiospecific isomers of fatty acyl chains will be distinguished using MS(2) and MS(3) fragment spectra. We applied the algorithm to the selection of 45 TAG standards and demonstrated that the molecular ions could be 100% correctly assigned. Therefore, the TIT software could be applied to TAG identification in complex biological samples such as mouse plasma extracts. Copyright © 2016 Elsevier B.V. All rights reserved.

Optical ID Tags for Secure Verification of Multispectral Visible and NIR Signatures

NASA Astrophysics Data System (ADS)

Pérez-Cabré, Elisabet; Millán, María S.; Javidi, Bahram

2008-04-01

We propose to combine information from visible (VIS) and near infrared (NIR) spectral bands to increase robustness on security systems and deter from unauthorized use of optical tags that permit the identification of a given person or object. The signature that identifies the element under surveillance will be only obtained by the appropriate combination of the visible content and the NIR data. The fully-phase encryption technique is applied to avoid an easy recognition of the resultant signature at the naked eye and an easy reproduction using conventional devices for imaging or scanning. The obtained complex-amplitude encrypted distribution is encoded on an identity (ID) tag. Spatial multiplexing of the encrypted signature allows us to build a distortion-invariant ID tag, so that remote authentication can be achieved even if the tag is captured under rotation or at different distances. We explore the possibility of using partial information of the encrypted distribution. Simulation results are provided and discussed.

Using RFID and PIT tags to Quantify Bedload Transport in the Oregon Coast Range

NASA Astrophysics Data System (ADS)

Sanfilippo, J.; Lancaster, S. T.

2017-12-01

Introducing the methods, issues, and data collection techniques and interpretation over one water year using RFID (radio frequency identification) and PIT (passive integrated transponder) tags in Oak Creek, near Corvallis Oregon. We constructed an RFID four-antenna array that runs off of a single radiofrequency reader via a multiplexer board. Using 4 grain sizes we tagged 990 individual rocks, roughly 250 in of each four size ranges (8-16mm, 16-32mm, 32-64mm, 64-128mm). Using 12 mm and 23 mm PIT tags during 1 water year the antennas logged 477 tracer passage events. To calculate bedload transport for each size range, at each antenna, interarrival times yield count rates when combined with grain size fractions of the bed and tracer concentrations, yield bedload transport for each size class. We calculated transport rates for five events of varying magnitude and found that PIT tag RFID method under predicts transport between 1 and 3 orders of magnitude.

Design of Distortion-Invariant Optical ID Tags for Remote Identification and Verification of Objects

NASA Astrophysics Data System (ADS)

Pérez-Cabré, Elisabet; Millán, María Sagrario; Javidi, Bahram

Optical identification (ID) tags [1] have a promising future in a number of applications such as the surveillance of vehicles in transportation, control of restricted areas for homeland security, item tracking on conveyor belts or other industrial environment, etc. More specifically, passive optical ID tag [1] was introduced as an optical code containing a signature (that is, a characteristic image or other relevant information of the object), which permits its real-time remote detection and identification. Since their introduction in the literature [1], some contributions have been proposed to increase their usefulness and robustness. To increase security and avoid counterfeiting, the signature was introduced in the optical code as an encrypted function [2-5] following the double-phase encryption technique [6]. Moreover, the design of the optical ID tag was done in such a way that tolerance to variations in scale and rotation was achieved [2-5]. To do that, the encrypted information was multiplexed and distributed in the optical code following an appropriate topology. Further studies were carried out to analyze the influence of different sources of noise. In some proposals [5, 7], the designed ID tag consists of two optical codes where the complex-valued encrypted signature was separately introduced in two real-valued functions according to its magnitude and phase distributions. This solution was introduced to overcome some difficulties in the readout of complex values in outdoors environments. Recently, the fully phase encryption technique [8] has been proposed to increase noise robustness of the authentication system.

Site-directed immobilization of a genetically engineered anti-methotrexate antibody via an enzymatically introduced biotin label significantly increases the binding capacity of immunoaffinity columns.

PubMed

Davenport, Kaitlynn R; Smith, Christopher A; Hofstetter, Heike; Horn, James R; Hofstetter, Oliver

2016-05-15

In this study, the effect of random vs. site-directed immobilization techniques on the performance of antibody-based HPLC columns was investigated using a single-domain camelid antibody (VHH) directed against methotrexate (MTX) as a model system. First, the high flow-through support material POROS-OH was activated with disuccinimidyl carbonate (DSC), and the VHH was bound in a random manner via amines located on the protein's surface. The resulting column was characterized by Frontal Affinity Chromatography (FAC). Then, two site-directed techniques were explored to increase column efficiency by immobilizing the antibody via its C-terminus, i.e., away from the antigen-binding site. In one approach, a tetra-lysine tail was added, and the antibody was immobilized onto DSC-activated POROS. In the second site-directed approach, the VHH was modified with the AviTag peptide, and a biotin-residue was enzymatically incorporated at the C-terminus using the biotin ligase BirA. The biotinylated antibody was subsequently immobilized onto NeutrAvidin-derivatized POROS. A comparison of the FAC analyses, which for all three columns showed excellent linearity (R(2)>0.999), revealed that both site-directed approaches yield better results than the random immobilization; the by far highest efficiency, however, was determined for the immunoaffinity column based on AviTag-biotinylated antibody. As proof of concept, all three columns were evaluated for quantification of MTX dissolved in phosphate buffered saline (PBS). Validation using UV-detection showed excellent linearity in the range of 0.04-12μM (R(2)>0.993). The lower limit of detection (LOD) and lower limit of quantification (LLOQ) were found to be independent of the immobilization strategy and were 40nM and 132nM, respectively. The intra- and inter-day precision was below 11.6%, and accuracy was between 90.7% and 112%. To the best of our knowledge, this is the first report of the AviTag-system in chromatography, and the first application of immunoaffinity chromatography for the analysis of MTX. Copyright © 2016 Elsevier B.V. All rights reserved.

Blockade of Tumor Cell TGF-Betas: A Strategy to Reverse Antiestrogen Resistance in Human Breast Cancer

DTIC Science & Technology

2002-01-01

the TM- FKHRL1 construct exhibited exclusive nuclear localization Cell Cycle Analysis by Flow Cytometry of the HA-tagged mutant under any experimental...distribution as measured by flow cytometry (Figure 8A). ALS AND METHODS. Consistent with its antiapoptotic effect, these results, addi- tion of TGFI3... flow cytometry . Under these conditions more than 95% of selected cells expressed GFP at the time of experiments. Immunoblot Analysis. Cells were

Diagnostics of Unseeded Air and Nitrogen Flows by Molecular Tagging

DTIC Science & Technology

2015-07-21

the polarization components across the FLEET line and the computed molecular density and dissociation fraction. Rayleigh Scattering Polarimetry V...Pol H Pol BPF HP CCM FLEET Emission Imaging Planar Rayleigh Scattering Rayleigh Scattering Polarimetry Density of molecules (normalized...the flow transport properties. This has been studied using Rayleigh scattering and Rayleigh scattering polarimetry . Figure 8 shows the evolution of

A model for estimating passive integrated transponder (PIT) tag antenna efficiencies for interval-specific emigration rates

USGS Publications Warehouse

Horton, G.E.; Dubreuil, T.L.; Letcher, B.H.

2007-01-01

Our goal was to understand movement and its interaction with survival for populations of stream salmonids at long-term study sites in the northeastern United States by employing passive integrated transponder (PIT) tags and associated technology. Although our PIT tag antenna arrays spanned the stream channel (at most flows) and were continuously operated, we are aware that aspects of fish behavior, environmental characteristics, and electronic limitations influenced our ability to detect 100% of the emigration from our stream site. Therefore, we required antenna efficiency estimates to adjust observed emigration rates. We obtained such estimates by testing a full-scale physical model of our PIT tag antenna array in a laboratory setting. From the physical model, we developed a statistical model that we used to predict efficiency in the field. The factors most important for predicting efficiency were external radio frequency signal and tag type. For most sampling intervals, there was concordance between the predicted and observed efficiencies, which allowed us to estimate the true emigration rate for our field populations of tagged salmonids. One caveat is that the model's utility may depend on its ability to characterize external radio frequency signals accurately. Another important consideration is the trade-off between the volume of data necessary to model efficiency accurately and the difficulty of storing and manipulating large amounts of data.

Downstream Migration of Masu Salmon Smolt at a Diversion Facility of Dam

NASA Astrophysics Data System (ADS)

Hayashida, K.; Nii, H.; Kasuga, K.; Watanabe, K.

2014-12-01

A diversion facility was installed on the upstream of Pirika Dam in Northern Japan that produced a downstream flow into the fishway, thus allowing the fish to migrate to the sea. On the other hand, if the flow rate in the river was more than 7.00 m 3/s (design flow rate of diversion facility), masu salmon smolt were concerned about accessing the dam reservoir, because the smolt can't migrate to the sea through the diversion facility unfortunately. Therefore, the downstream migration of smolt was investigated around the diversion facility. The PIT tag system and radio transmitters as the biotelemetry were used to determine 1) whether masu salmon smolt were able to migrate downstream through the diversion facility and fishway at Pirika Dam, 2) when the smolt started to migrate downstream, 3) whether the downstream migration of smolt were affected by the flow increase in the river. It was clarified that 88% of the smolt were able to enter the diversion facility, and then 81% of the smolt were able to access the fishway. It was also clarified that smolt downstream migration had two peaks in a day (5:00 and 18:00). During the study period, although the flow rate was in the 2.21 m3/s to 30.44 m3/s range (average 6.70 m3/s), it was revealed that the diversion facility has a satisfactory function for the downstream migration of smolt as presented above. The survey clarified the downstream migration behavior of masu salmon by using two types of biotelemetry equipment. PIT tag and radio transmitter were found to be very effective in tracking the behavior of small fish such as smolt. PIT tags, in particular, require very little operating cost, because once they are inserted in the fish, they do not need human labor for tracking. It is desirable to actively introduce the biotelemetry as tracking equipment when surveying the fish migration in the river.

Progress in tagged neutron beams for cargo inspections

NASA Astrophysics Data System (ADS)

Pesente, S.; Nebbia, G.; Viesti, G.; Daniele, F.; Fabris, D.; Lunardon, M.; Moretto, S.; Nad, K.; Sudac, D.; Valkovic, V.

2007-08-01

The use of neutron beams produced via the D + T reaction and tagged by the associated particle technique has been recently applied to cargo container inspections. In the EURITRACK project, a portable sealed-tube neutron generator has been designed and built to deliver 14 MeV neutron beams tagged by a matrix of 64 YAP:Ce alpha-particle detectors read by a multi-anode HAMAMATSU H8500 Photomultiplier Tube. The performances of this alpha-particle detector have been determined as a function of the count rate at the Rudjer Boskovic Institute, Zagreb (Croatia). Moreover, tests of the final detector operated inside the sealed-tube neutron generator are fully satisfactory.

Direct match data flow machine apparatus and process for data driven computing

DOEpatents

Davidson, G.S.; Grafe, V.G.

1997-08-12

A data flow computer and method of computing are disclosed which utilizes a data driven processor node architecture. The apparatus in a preferred embodiment includes a plurality of First-In-First-Out (FIFO) registers, a plurality of related data flow memories, and a processor. The processor makes the necessary calculations and includes a control unit to generate signals to enable the appropriate FIFO register receiving the result. In a particular embodiment, there are three FIFO registers per node: an input FIFO register to receive input information form an outside source and provide it to the data flow memories; an output FIFO register to provide output information from the processor to an outside recipient; and an internal FIFO register to provide information from the processor back to the data flow memories. The data flow memories are comprised of four commonly addressed memories. A parameter memory holds the A and B parameters used in the calculations; an opcode memory holds the instruction; a target memory holds the output address; and a tag memory contains status bits for each parameter. One status bit indicates whether the corresponding parameter is in the parameter memory and one status but to indicate whether the stored information in the corresponding data parameter is to be reused. The tag memory outputs a ``fire`` signal (signal R VALID) when all of the necessary information has been stored in the data flow memories, and thus when the instruction is ready to be fired to the processor. 11 figs.

Data flow machine for data driven computing

DOEpatents

Davidson, G.S.; Grafe, V.G.

1988-07-22

A data flow computer and method of computing is disclosed which utilizes a data driven processor node architecture. The apparatus in a preferred embodiment includes a plurality of First-In-First-Out (FIFO) registers, a plurality of related data flow memories, and a processor. The processor makes the necessary calculations and includes a control unit to generate signals to enable the appropriate FIFO register receiving the result. In a particular embodiment, there are three FIFO registers per node: an input FIFO register to receive input information from an outside source and provide it to the data flow memories; an output FIFO register to provide output information from the processor to an outside recipient; and an internal FIFO register to provide information from the processor back to the data flow memories. The data flow memories are comprised of four commonly addressed memories. A parameter memory holds the A and B parameters used in the calculations; an opcode memory holds the instruction; a target memory holds the output address; and a tag memory contains status bits for each parameter. One status bit indicates whether the corresponding parameter is in the parameter memory and one status bit to indicate whether the stored information in the corresponding data parameter is to be reused. The tag memory outputs a ''fire'' signal (signal R VALID) when all of the necessary information has been stored in the data flow memories, and thus when the instruction is ready to be fired to the processor. 11 figs.

Data flow machine for data driven computing

DOEpatents

Davidson, George S.; Grafe, Victor G.

1995-01-01

A data flow computer which of computing is disclosed which utilizes a data driven processor node architecture. The apparatus in a preferred embodiment includes a plurality of First-In-First-Out (FIFO) registers, a plurality of related data flow memories, and a processor. The processor makes the necessary calculations and includes a control unit to generate signals to enable the appropriate FIFO register receiving the result. In a particular embodiment, there are three FIFO registers per node: an input FIFO register to receive input information form an outside source and provide it to the data flow memories; an output FIFO register to provide output information from the processor to an outside recipient; and an internal FIFO register to provide information from the processor back to the data flow memories. The data flow memories are comprised of four commonly addressed memories. A parameter memory holds the A and B parameters used in the calculations; an opcode memory holds the instruction; a target memory holds the output address; and a tag memory contains status bits for each parameter. One status bit indicates whether the corresponding parameter is in the parameter memory and one status but to indicate whether the stored information in the corresponding data parameter is to be reused. The tag memory outputs a "fire" signal (signal R VALID) when all of the necessary information has been stored in the data flow memories, and thus when the instruction is ready to be fired to the processor.

Direct match data flow machine apparatus and process for data driven computing

DOEpatents

Davidson, George S.; Grafe, Victor Gerald

1997-01-01

A data flow computer and method of computing is disclosed which utilizes a data driven processor node architecture. The apparatus in a preferred embodiment includes a plurality of First-In-First-Out (FIFO) registers, a plurality of related data flow memories, and a processor. The processor makes the necessary calculations and includes a control unit to generate signals to enable the appropriate FIFO register receiving the result. In a particular embodiment, there are three FIFO registers per node: an input FIFO register to receive input information form an outside source and provide it to the data flow memories; an output FIFO register to provide output information from the processor to an outside recipient; and an internal FIFO register to provide information from the processor back to the data flow memories. The data flow memories are comprised of four commonly addressed memories. A parameter memory holds the A and B parameters used in the calculations; an opcode memory holds the instruction; a target memory holds the output address; and a tag memory contains status bits for each parameter. One status bit indicates whether the corresponding parameter is in the parameter memory and one status but to indicate whether the stored information in the corresponding data parameter is to be reused. The tag memory outputs a "fire" signal (signal R VALID) when all of the necessary information has been stored in the data flow memories, and thus when the instruction is ready to be fired to the processor.

Direct match data flow memory for data driven computing

DOEpatents

Davidson, George S.; Grafe, Victor Gerald

1997-01-01

A data flow computer and method of computing is disclosed which utilizes a data driven processor node architecture. The apparatus in a preferred embodiment includes a plurality of First-In-First-Out (FIFO) registers, a plurality of related data flow memories, and a processor. The processor makes the necessary calculations and includes a control unit to generate signals to enable the appropriate FIFO register receiving the result. In a particular embodiment, there are three FIFO registers per node: an input FIFO register to receive input information form an outside source and provide it to the data flow memories; an output FIFO register to provide output information from the processor to an outside recipient; and an internal FIFO register to provide information from the processor back to the data flow memories. The data flow memories are comprised of four commonly addressed memories. A parameter memory holds the A and B parameters used in the calculations; an opcode memory holds the instruction; a target memory holds the output address; and a tag memory contains status bits for each parameter. One status bit indicates whether the corresponding parameter is in the parameter memory and one status bit to indicate whether the stored information in the corresponding data parameter is to be reused. The tag memory outputs a "fire" signal (signal R VALID) when all of the necessary information has been stored in the data flow memories, and thus when the instruction is ready to be fired to the processor.

Direct match data flow memory for data driven computing

DOEpatents

Davidson, G.S.; Grafe, V.G.

1997-10-07

A data flow computer and method of computing is disclosed which utilizes a data driven processor node architecture. The apparatus in a preferred embodiment includes a plurality of First-In-First-Out (FIFO) registers, a plurality of related data flow memories, and a processor. The processor makes the necessary calculations and includes a control unit to generate signals to enable the appropriate FIFO register receiving the result. In a particular embodiment, there are three FIFO registers per node: an input FIFO register to receive input information form an outside source and provide it to the data flow memories; an output FIFO register to provide output information from the processor to an outside recipient; and an internal FIFO register to provide information from the processor back to the data flow memories. The data flow memories are comprised of four commonly addressed memories. A parameter memory holds the A and B parameters used in the calculations; an opcode memory holds the instruction; a target memory holds the output address; and a tag memory contains status bits for each parameter. One status bit indicates whether the corresponding parameter is in the parameter memory and one status bit to indicate whether the stored information in the corresponding data parameter is to be reused. The tag memory outputs a ``fire`` signal (signal R VALID) when all of the necessary information has been stored in the data flow memories, and thus when the instruction is ready to be fired to the processor. 11 figs.

Innovative techniques for analyzing the three-dimensional behavioral results from acoustically tagged fish

NASA Astrophysics Data System (ADS)

Steig, Tracey W.; Timko, Mark A.

2005-04-01

Acoustic tags were used to monitor the swimming patterns of downstream migrating salmon smolts approaching various dams on the Columbia River, USA. Downstream migrating yearling chinook (Oncorhynchus tshawytscha), steelhead (Oncorhynchus mykiss), sockeye (Oncorhynchus nerka), and sub-yearling chinook smolts were surgically implanted with acoustic tags. Fish were tracked in three-dimensions as they approached and passed into the turbine intakes, spillways, and surface bypass channel entrances at the dams during the 2004 spring and summer outmigrations. A number of advances in the analysis techniques and software have been made over the past few years. Some of these improvements include the development of various fish density algorithms, stream trace modeling analysis, and advances of three-dimensional animation programs. Three-dimensional tracks of fish approaching the turbine intakes, spillways, and surface bypass channel entrances will be presented. Concentrations of fish passage will be presented as three-dimensional fish densities superimposed over dam structures. Stream trace modeling animation will be presented showing predicted fish passage routes.

Radio-transmitters do not affect seasonal productivity of female Golden-winged Warblers

USGS Publications Warehouse

Streby, Henry M.; Peterson, Sean M.; Gesmundo, Callie; Johnson, Michael K.; Fish, Alexander C.; Lehman, Justin A.; Andersen, David E.

2013-01-01

Investigating the potential effects of handling and marking techniques on study animals is important for correct interpretation of research results and to effect progress in data-collection methods. Few investigators have compared the reproductive output of radio-tagged and non-radio-tagged songbirds, and no one to date has examined the possible effect of radio-tagging adult songbirds on the survival of their fledglings. In 2011 and 2012, we compared several parameters of reproductive output of two groups of female Golden-winged Warblers (Vermivora chrysoptera) breeding in Minnesota, including 45 females with radio-transmitters and 73 females we did not capture, handle, or mark. We found no difference between groups in clutch sizes, hatching success, brood sizes, length of incubation and nestling stages, fledging success, number of fledglings, or survival of fledglings to independence. Thus, radio-tags had no measurable impact on the productivity of female Golden-winged Warblers. Our results build upon previous studies where investigators have reported no effects of radio-tagging on the breeding parameters of songbirds by also demonstrating no effect of radio-tagging through the post-fledging period and, therefore, the entire breeding season.

The Interaction between Logjams, Channel Evolution, and Sports Fisheries on a Dam Regulated Low Gradient River.

NASA Astrophysics Data System (ADS)

Schenk, E.; Hupp, C. R.; Moulin, B.

2014-12-01

The purpose of our study was to determine the interaction between in-stream large wood (LW), bank erosion, and sports fisheries in the 210 river kilometer (km) Coastal Plain segment of the dam-regulated Roanoke River, North Carolina. Methods included collecting background geomorphic data including a 200 km channel geometry survey and measurements from 701 bank erosion pins at 36 cross-sections over 132 km. LW concentrations were evaluated over a 177 km reach using georeferenced aerial video taken during regulated low flow (56 m3/s). LW transport was measured using 290 radio tagged LW pieces (mean diameter = 35.0 cm, length = 9.3 m) installed between 2008 and 2010. Largemouth bass (Micropterus salmoides) were surveyed in 2010 at 29 sites using a boat mounted electroshock unit. The abundance of LW in logjams was 59 pieces/km and these were concentrated (21.5 logjams/km) in an actively eroding reach with relatively high sinuosity, high local LW production rates, and narrow channel widths. Most jams (70%) are available nearly year round as aquatic habitat, positioned either on the lower bank or submerged at low-water flows. The actively eroding reach is adjusting to upstream dam regulation by channel widening. The channel upstream of this reach has widened and stabilized while the channel downstream of the eroding reach is still relatively narrow but with lower bank erosion rates. Repeat surveys of radio tagged LW determined that transport was common throughout the study area despite dam regulation and a low channel gradient (0.0016). The mean distance travelled by a radio tagged piece of LW was 11.9 km with a maximum of 101 km (84 tags moved, 96 stationary, 110 not found). Radio tagged LW that moved during the study was found at low flow either in logjams (44%), as individual LW (43%), or submerged mid-channel (14%). Largemouth bass biomass density (g/hr effort) was highest in the actively eroding reach where logjams were most common. Our results support the hypothesis that channel evolution processes control bank stability and complexity that in turn control logjam frequency. Areas with higher concentrations of logjams have larger and more largemouth bass, a valued sports fish.

Infrared tag and track technique

DOEpatents

Partin, Judy K.; Stone, Mark L.; Slater, John; Davidson, James R.

2007-12-04

A method of covertly tagging an object for later tracking includes providing a material capable of at least one of being applied to the object and being included in the object, which material includes deuterium; and performing at least one of applying the material to the object and including the material in the object in a manner in which in the appearance of the object is not changed, to the naked eye.

GSyellow, a Multifaceted Tag for Functional Protein Analysis in Monocot and Dicot Plants.

PubMed

Besbrugge, Nienke; Van Leene, Jelle; Eeckhout, Dominique; Cannoot, Bernard; Kulkarni, Shubhada R; De Winne, Nancy; Persiau, Geert; Van De Slijke, Eveline; Bontinck, Michiel; Aesaert, Stijn; Impens, Francis; Gevaert, Kris; Van Damme, Daniel; Van Lijsebettens, Mieke; Inzé, Dirk; Vandepoele, Klaas; Nelissen, Hilde; De Jaeger, Geert

2018-06-01

The ability to tag proteins has boosted the emergence of generic molecular methods for protein functional analysis. Fluorescent protein tags are used to visualize protein localization, and affinity tags enable the mapping of molecular interactions by, for example, tandem affinity purification or chromatin immunoprecipitation. To apply these widely used molecular techniques on a single transgenic plant line, we developed a multifunctional tandem affinity purification tag, named GS yellow , which combines the streptavidin-binding peptide tag with citrine yellow fluorescent protein. We demonstrated the versatility of the GS yellow tag in the dicot Arabidopsis ( Arabidopsis thaliana ) using a set of benchmark proteins. For proof of concept in monocots, we assessed the localization and dynamic interaction profile of the leaf growth regulator ANGUSTIFOLIA3 (AN3), fused to the GS yellow tag, along the growth zone of the maize ( Zea mays ) leaf. To further explore the function of ZmAN3, we mapped its DNA-binding landscape in the growth zone of the maize leaf through chromatin immunoprecipitation sequencing. Comparison with AN3 target genes mapped in the developing maize tassel or in Arabidopsis cell cultures revealed strong conservation of AN3 target genes between different maize tissues and across monocots and dicots, respectively. In conclusion, the GS yellow tag offers a powerful molecular tool for distinct types of protein functional analyses in dicots and monocots. As this approach involves transforming a single construct, it is likely to accelerate both basic and translational plant research. © 2018 American Society of Plant Biologists. All rights reserved.

Multiparameter Flowfield Measurements in High-Pressure, Cryogenic Environments Using Femtosecond Lasers

NASA Technical Reports Server (NTRS)

Burns, Ross A.; Danehy, Paul M.; Peters, Christopher J.

2016-01-01

Femtosecond laser electronic excitation tagging (FLEET) and Rayleigh scattering (RS) from a femtosecond laser are demonstrated in the NASA Langley 0.3-m Transonic Cryogenic Tunnel (TCT). The measured signals from these techniques are examined for their thermodynamic dependencies in pure nitrogen. The FLEET signal intensity and signal lifetimes are found to scale primarily with the gas density, as does the RS signal. Several models are developed, which capture these physical behaviors. Notably, the FLEET and Rayleigh scattering intensities scale linearly with the flow density, while the FLEET signal decay rates are a more complex function of the thermodynamic state of the gas. The measurement of various flow properties are demonstrated using these techniques. While density was directly measured from the signal intensities and FLEET signal lifetime, temperature and pressure were measured using the simultaneous FLEET velocity measurements while assuming the flow had a constant total enthalpy. Measurements of density, temperature, and pressure from the FLEET signal are made with accuracies as high as 5.3 percent, 0.62 percent, and 6.2 percent, respectively, while precisions were approximately 10 percent, 0.26 percent, and 11 percent for these same quantities. Similar measurements of density from Rayleigh scattering showed an overall accuracy of 3.5 percent and a precision of 10.2 percent over a limited temperature range (T greater than 195 K). These measurements suggest a high degree of utility at using the femtosecond-laser based diagnostics for making multiparameter measurements in high-pressure, cryogenic environments such as large-scale TCT facilities.

Dichroic subjettiness ratios to distinguish colour flows in boosted boson tagging

NASA Astrophysics Data System (ADS)

Salam, Gavin P.; Schunk, Lais; Soyez, Gregory

2017-03-01

N-subjettiness ratios are in wide use for tagging heavy boosted objects, in particular the ratio of 2-subjettiness to 1-subjettiness for tagging boosted electroweak bosons. In this article we introduce a new, dichroic ratio, which uses different regions of a jet to determine the two subjettiness measures, emphasising the hard substructure for the 1-subjettiness and the full colour radiation pattern for the 2-subjettiness. Relative to existing N -subjettiness ratios, the dichroic extension, combined with SoftDrop (pre-)grooming, makes it possible to increase the ultimate signal significance by about 25% (for 2 TeV jets), or to reduce non-perturbative effects by a factor of 2-3 at 50% signal efficiency while maintaining comparable background rejection. We motivate the dichroic approach through the study of Lund diagrams, supplemented with resummed analytical calculations.

Aeration strategy for biofilm cultivation of the microalga Scenedesmus dimorphus.

PubMed

Ji, Chunli; Wang, Junfeng; Liu, Tianzhong

2015-10-01

Biofilm cultivation of microalgae may be useful for biofuel production. However, many aspects for this cultivation method have not been well assessed. Accordingly, aeration strategy for biofilm cultivation of Scenedesmus dimorphus has been explored. Biomass, lipid and triacylglycerol (TAG) productivity in increased S. dimorphus as the CO2 concentration increased within 0.038-0.5% and kept constant with further increases. The biomass, lipid and TAG productivity increased with the speed increasing and an obvious threshold point was observed at 6.6 ml(-2) min(-1). The lipid and TAG content was unaffected by the aeration rate. Both the CO2 concentration as well as aeration speed affected the growth of S. dimorphus in biofilm cultivation. The optimized aeration strategy for biofilm cultivation was continuous air flow enriched with 1% CO2 (v/v) at 6.6 ml(-2) min(-1).

A comparison of conventional capture versus PIT reader techniques for estimating survival and capture probabilities of big brown bats (Eptesicus fuscus)

USGS Publications Warehouse

Ellison, L.E.; O'Shea, T.J.; Neubaum, D.J.; Neubaum, M.A.; Pearce, R.D.; Bowen, R.A.

2007-01-01

We compared conventional capture (primarily mist nets and harp traps) and passive integrated transponder (PIT) tagging techniques for estimating capture and survival probabilities of big brown bats (Eptesicus fuscus) roosting in buildings in Fort Collins, Colorado. A total of 987 female adult and juvenile bats were captured and marked by subdermal injection of PIT tags during the summers of 2001-2005 at five maternity colonies in buildings. Openings to roosts were equipped with PIT hoop-style readers, and exit and entry of bats were passively monitored on a daily basis throughout the summers of 2002-2005. PIT readers 'recaptured' adult and juvenile females more often than conventional capture events at each roost. Estimates of annual capture probabilities for all five colonies were on average twice as high when estimated from PIT reader data (P?? = 0.93-1.00) than when derived from conventional techniques (P?? = 0.26-0.66), and as a consequence annual survival estimates were more precisely estimated when using PIT reader encounters. Short-term, daily capture estimates were also higher using PIT readers than conventional captures. We discuss the advantages and limitations of using PIT tags and passive encounters with hoop readers vs. conventional capture techniques for estimating these vital parameters in big brown bats. ?? Museum and Institute of Zoology PAS.

Comparison of magnetic carboxymethyl chitosan nanoparticles and cation exchange resin for the efficient purification of lysine-tagged small ubiquitin-like modifier protease.

PubMed

Li, Junhua; Zhang, Yang; Shen, Fei; Yang, Yanjun

2012-10-15

A fusion tag that can be purified by the cheap ion-exchanger based on the ionic binding force may provide a cost-effective scheme over other affinity fusion tags. Small ubiquitin-like modifier (SUMO) protease derived from Saccharomyces cerevisiae was fused with a poly lysine tag containing 10 lysine residues at its C-terminus and then expressed in Escherichia coli. The ionic binding force provided by the ploy lysine tag allowed the selective recovery of the small ubiquitin-like modifier protease from recombinant E. coli cell extracts. A preliminary comparative study of the adsorption and elution of poly lysine tagged SUMO protease on Amberlite Cobalamion and magnetite carboxymethyl chitosan nanoparticles was performed. Amberlite Cobalamion and magnetite nanoparticles had the similar elution profile due to the common functional groups - carboxyl groups. The maximum dynamic adsorption capacity of Amberlite Cobalamion and magnetite nanoparticles reached 36.8 and 211.4 mg/g, respectively. The lysine-tagged protease can be simply purified by magnetite nanoparticles from cell extracts with higher purity than that by Amberlite Cobalamion. The superparamagnetic nanoparticles possess the advantages of highly specific, fast and excellent binding of a larger amount of lysine tagged SUMO modifier protease, and it is also easier to separate from the crude biological process liquors compared with the conventional separation techniques of polycationic amino acids fusion proteins. Copyright © 2012 Elsevier B.V. All rights reserved.

77 FR 57554 - Endangered and Threatened Species; Reopening of Public Comment Period on Proposed Endangered...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-09-18

... and journal articles are: (1) Preliminary Results from Photo-identification and Satellite- tagging of... Mechanisms of Gene Flow within Island- associated False Killer Whales around the Hawaiian Archipelago, by...

DNA-tagged Microparticles for Tracing Water Flows and Travel Times in Natural Systems: The First results from Controlled Laboratory Experiments

NASA Astrophysics Data System (ADS)

Bogaard, T.; Bandyopadhyay, S.; Foppen, J. W.

2017-12-01

Societal demand for water safety is continuously increasing, being it resilient against flood/droughts, clean water for ecosystems, recreation or safe drinking water. Robust methods to measure temporal and spatial patterns of water and contaminant pathways are still lacking. Our research project aims to develop and apply (1) innovative, robust, and environmental-friendly silica-protected iron oxide micro-particles tagged with artificial DNA to trace contaminant movement and travel times of water in natural systems and (2) an innovative coupled model approach to capture dynamics in hydrological pathways and their effects on water quality. The exceptional property of DNA-tagging is the infinite number of unique tracers that can be produced and their detectability at extreme low concentrations. The advantage of the iron-core of the particle is the magnetic harvesting of the particles from water-samples. Such tracers are thought to give the water sector a unique tool for in-situ mapping of transport of contaminants and pathogenic microorganisms in water systems. However, the characteristics of the particle like magnetic property of the iron-core and surface potential of the silica layer, are of key importance for the behaviour of the particle in surface water and in soils. Furthermore, the application of such micro-particles requires strict protocols for the experiment, sampling and laboratory handling which are currently not available. We used two different types of silica-protected DNA-tagged micro-particles. We performed batch, column and flow experiments to assess the behaviour of the particles. We will present the first results of the controlled laboratory experiments for hydrological tracing. We will discuss the results and link it to the differences in particles design. Furthermore, we will draw conclusions and discuss knowledge gaps for future application of silica-protected DNA-tagged micro-particles in hydrological research.

Monitoring and Evaluation: Statistical Support for Life-cycle Studies, Annual Report 2003.

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

Skalski, John

2003-11-01

The ongoing mission of this project is the development of statistical tools for analyzing fisheries tagging data in the most precise and appropriate manner possible. This mission also includes providing statistical guidance on the best ways to design large-scale tagging studies. This mission continues because the technologies for conducting fish tagging studies continuously evolve. In just the last decade, fisheries biologists have seen the evolution from freeze-brands and coded wire tags (CWT) to passive integrated transponder (PIT) tags, balloon-tags, radiotelemetry, and now, acoustic-tags. With each advance, the technology holds the promise of more detailed and precise information. However, the technologymore » for analyzing and interpreting the data also becomes more complex as the tagging techniques become more sophisticated. The goal of the project is to develop the analytical tools in parallel with the technical advances in tagging studies, so that maximum information can be extracted on a timely basis. Associated with this mission is the transfer of these analytical capabilities to the field investigators to assure consistency and the highest levels of design and analysis throughout the fisheries community. Consequently, this project provides detailed technical assistance on the design and analysis of tagging studies to groups requesting assistance throughout the fisheries community. Ideally, each project and each investigator would invest in the statistical support needed for the successful completion of their study. However, this is an ideal that is rarely if every attained. Furthermore, there is only a small pool of highly trained scientists in this specialized area of tag analysis here in the Northwest. Project 198910700 provides the financial support to sustain this local expertise on the statistical theory of tag analysis at the University of Washington and make it available to the fisheries community. Piecemeal and fragmented support from various agencies and organizations would be incapable of maintaining a center of expertise. The mission of the project is to help assure tagging studies are designed and analyzed from the onset to extract the best available information using state-of-the-art statistical methods. The overarching goals of the project is to assure statistically sound survival studies so that fish managers can focus on the management implications of their findings and not be distracted by concerns whether the studies are statistically reliable or not. Specific goals and objectives of the study include the following: (1) Provide consistent application of statistical methodologies for survival estimation across all salmon life cycle stages to assure comparable performance measures and assessment of results through time, to maximize learning and adaptive management opportunities, and to improve and maintain the ability to responsibly evaluate the success of implemented Columbia River FWP salmonid mitigation programs and identify future mitigation options. (2) Improve analytical capabilities to conduct research on survival processes of wild and hatchery chinook and steelhead during smolt outmigration, to improve monitoring and evaluation capabilities and assist in-season river management to optimize operational and fish passage strategies to maximize survival. (3) Extend statistical support to estimate ocean survival and in-river survival of returning adults. Provide statistical guidance in implementing a river-wide adult PIT-tag detection capability. (4) Develop statistical methods for survival estimation for all potential users and make this information available through peer-reviewed publications, statistical software, and technology transfers to organizations such as NOAA Fisheries, the Fish Passage Center, US Fish and Wildlife Service, US Geological Survey (USGS), US Army Corps of Engineers (USACE), Public Utility Districts (PUDs), the Independent Scientific Advisory Board (ISAB), and other members of the Northwest fisheries community. (5) Provide and maintain statistical software for tag analysis and user support. (6) Provide improvements in statistical theory and software as requested by user groups. These improvements include extending software capabilities to address new research issues, adapting tagging techniques to new study designs, and extending the analysis capabilities to new technologies such as radio-tags and acoustic-tags.« less

Simplifications of the RELIEF flow tagging system for laboratory use. [Raman Excitation plus Laser Induced Electronic Fluorescence

NASA Technical Reports Server (NTRS)

Lempert, Walter R.; Zhang, Boying; Miles, Richard B.; Diskin, Glenn

1991-01-01

The use of an O2:He stimulated Raman cell to generate the Stokes beam for the Raman vibrational pumping step of the RELIEF (Raman Excitation plus Laser-Induced Electronic Fluorescence) flow tagging method is reported. Use of the Raman cell rather than a dye laser provides pump and Stokes beams which are automatically frequency matched and temporally and spatially overlapped. The Nd:YAG pump laser is operated multilongitudinal mode, which eliminates the need for injection seeding, resulting in decreased operation complexity and improved stability with respect to acoustic noise. Results are presented for 1st Stokes conversion efficiency and stimulated Brillouin backscattering loss and are compared to the case of pure O2. Scanning CARS measurements of the Q-branch lineshape for both pure O2 and the O2:He mixture are also presented.

Precision of FLEET Velocimetry Using High-Speed CMOS Camera Systems

NASA Technical Reports Server (NTRS)

Peters, Christopher J.; Danehy, Paul M.; Bathel, Brett F.; Jiang, Naibo; Calvert, Nathan D.; Miles, Richard B.

2015-01-01

Femtosecond laser electronic excitation tagging (FLEET) is an optical measurement technique that permits quantitative velocimetry of unseeded air or nitrogen using a single laser and a single camera. In this paper, we seek to determine the fundamental precision of the FLEET technique using high-speed complementary metal-oxide semiconductor (CMOS) cameras. Also, we compare the performance of several different high-speed CMOS camera systems for acquiring FLEET velocimetry data in air and nitrogen free-jet flows. The precision was defined as the standard deviation of a set of several hundred single-shot velocity measurements. Methods of enhancing the precision of the measurement were explored such as digital binning (similar in concept to on-sensor binning, but done in post-processing), row-wise digital binning of the signal in adjacent pixels and increasing the time delay between successive exposures. These techniques generally improved precision; however, binning provided the greatest improvement to the un-intensified camera systems which had low signal-to-noise ratio. When binning row-wise by 8 pixels (about the thickness of the tagged region) and using an inter-frame delay of 65 microseconds, precisions of 0.5 meters per second in air and 0.2 meters per second in nitrogen were achieved. The camera comparison included a pco.dimax HD, a LaVision Imager scientific CMOS (sCMOS) and a Photron FASTCAM SA-X2, along with a two-stage LaVision HighSpeed IRO intensifier. Excluding the LaVision Imager sCMOS, the cameras were tested with and without intensification and with both short and long inter-frame delays. Use of intensification and longer inter-frame delay generally improved precision. Overall, the Photron FASTCAM SA-X2 exhibited the best performance in terms of greatest precision and highest signal-to-noise ratio primarily because it had the largest pixels.

Distribution and Joint Fish-Tag Survival of Juvenile Chinook Salmon Migrating through the Sacramento-San Joaquin River Delta, California, 2008

USGS Publications Warehouse

Holbrook, Christopher M.; Perry, Russell W.; Adams, Noah S.

2009-01-01

Acoustic telemetry was used to obtain the movement histories of 915 juvenile fall-run Chinook salmon (Oncorhynchus tshawytscha) through the lower San Joaquin River and Sacramento-San Joaquin Delta, California, in 2008. Data were analyzed within a release-recapture framework to estimate survival, route distribution, and detection probabilities among three migration pathways through the Delta. The pathways included the primary route through the San Joaquin River and two less direct routes (Old River and Turner Cut). Strong inferences about survival were limited by premature tag failure, but estimates of fish distribution among migration routes should be unaffected by tag failure. Based on tag failure tests (N = 66 tags), we estimated that only 55-78 percent of the tags used in this study were still functioning when the last fish was detected exiting the study area 15 days after release. Due to premature tag failure, our 'survival' estimates represent the joint probability that both the tag and fish survived, not just survival of fish. Low estimates of fish-tag survival could have been caused by fish mortality or fish travel times that exceeded the life of the tag, but we were unable to differentiate between the two. Fish-tag survival through the Delta (from Durham Ferry to Chipps Island by all routes) ranged from 0.05 +or- 0.01 (SE) to 0.06 +or- 0.01 between the two weekly release groups. Among the three migration routes, fish that remained in the San Joaquin River exhibited the highest joint fish-tag survival (0.09 +or- 0.02) in both weeks, but only 22-33 percent of tagged fish used this route, depending on the week of release. Only 4-10 percent (depending on week) of tagged fish traveled through Turner Cut, but no tagged fish that used this route were detected exiting the Delta. Most fish (63-68 percent, depending on week of release) migrated through Old River, but fish-tag survival through this route (0.05 +or- 0.01) was only about one-half that of fish that remained in the San Joaquin River. Once tagged fish entered Old River, only fish collected at two large water conveyance projects and transported through the Delta by truck were detected exiting the Delta, suggesting that this route was the only successful migration pathway for fish that entered Old River. The rate of entrainment of tagged juvenile salmon into Old River was similar to the fraction of San Joaquin River discharge flowing into Old River, which averaged 63 percent but varied tidally and ranged from 33 to 100 percent daily. Although improvements in transmitter battery life are clearly needed, this information will help guide the development of future research and monitoring efforts in this system.

OPTIMIZATION OF THE WASH-OFF METHOD FOR MEASURING AEROSOL CONCENTRATIONS

EPA Science Inventory

Using the fluorescence-washing technique, oleic acid particles tagged with uranine were extracted and analyzed fluorometrically. The possible sources of errors in the technique were evaluated in this study. First, the sensitivity of uranine fluorescence in different solutions ...

Microfluidic immunomagnetic cell separation from whole blood.

PubMed

Bhuvanendran Nair Gourikutty, Sajay; Chang, Chia-Pin; Puiu, Poenar Daniel

2016-02-01

Immunomagnetic-based separation has become a viable technique for the separation of cells and biomolecules. Here we report on the design and analysis of a simple and efficient microfluidic device for high throughput and high efficiency capture of cells tagged with magnetic particles. This is made possible by using a microfluidic chip integrated with customized arrays of permanent magnets capable of creating large magnetic field gradients, which determine the effective capturing of the tagged cells. This method is based on manipulating the cells which are under the influence of a combination of magnetic and fluid dynamic forces in a fluid under laminar flow through a microfluidic chip. A finite element analysis (FEA) model is developed to analyze the cell separation process and predict its behavior, which is validated subsequently by the experimental results. The magnetic field gradients created by various arrangements of magnetic arrays have been simulated using FEA and the influence of these field gradients on cell separation has been studied with the design of our microfluidic chip. The proof-of-concept for the proposed technique is demonstrated by capturing white blood cells (WBCs) from whole human blood. CD45-conjugated magnetic particles were added into whole blood samples to label WBCs and the mixture was flown through our microfluidic device to separate the labeled cells. After the separation process, the remaining WBCs in the elute were counted to determine the capture efficiency, and it was found that more than 99.9% WBCs have been successfully separated from whole blood. The proposed design can be used for positive selection as well as for negative enrichment of rare cells. Copyright © 2015 Elsevier B.V. All rights reserved.

Low-cost passive UHF RFID tags on paper substrates

NASA Astrophysics Data System (ADS)

Sajal, Sayeed Zebaul Haque

To reduce the significant cost in the widespread deployment of UHF radio frequency identification (RFID) systems, an UHF RFID tag design is presented on paper substrates. The design is based on meander-line miniaturization techniques and open complementary split ring resonator (OCSRR) elements that reduce required conducting materials by 30%. Another passive UHF RFID tag is designed to sense the moisture based on the antenna's polarization. An inexpensive paper substrate and copper layer are used for flexibility and low-cost. The key characteristic of this design is the sensitivity of the antenna's polarization on the passive RFID tag to the moisture content in the paper substrate. In simulations, the antenna is circularly-polarized when the substrate is dry and is linearly-polarized when the substrate is wet. It was shown that the expected read-ranges and desired performance could be achieved reducing the over-all cost of the both designs.

A novel helper phage for HaloTag-mediated co-display of enzyme and substrate on phage.

PubMed

Delespaul, Wouter; Peeters, Yves; Herdewijn, Piet; Robben, Johan

2015-05-01

Phage display is an established technique for the molecular evolution of peptides and proteins. For the selection of enzymes based on catalytic activity however, simultaneous coupling of an enzyme and its substrate to the phage surface is required. To facilitate this process of co-display, we developed a new helper phage displaying HaloTag, a modified haloalkane dehalogenase that binds specifically and covalently to functionalized haloalkane ligands. The display of functional HaloTag was demonstrated by capture on streptavidin-coated magnetic beads, after coupling a biotinylated haloalkane ligand, or after on-phage extension of a DNA oligonucleotide primer with a biotinylated nucleotide by phi29 DNA polymerase. We also achieved co-display of HaloTag and phi29 DNA polymerase, thereby opening perspectives for the molecular evolution of this enzyme (and others) towards new substrate specificities. Copyright © 2015 Elsevier Inc. All rights reserved.

Performance Assessment of Bi-Directional Knotless Tissue-Closure Devices in Juvenile Chinook Salmon Surgically Implanted with Acoustic Transmitters, 2009 - Final Report

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

Woodley, Christa M.; Wagner, Katie A.; Bryson, Amanda J.

2012-11-09

The purpose of this report is to assess the performance of bi-directional knotless tissue-closure devices for use in tagging juvenile salmon. This study is part of an ongoing effort at Pacific Northwest National Laboratory (PNNL) to reduce unwanted effects of tags and tagging procedures on the survival and behavior of juvenile salmonids, by assessing and refining suturing techniques, suture materials, and tag burdens. The objective of this study was to compare the performance of the knotless (barbed) suture, using three different suture patterns (treatments: 6-point, Wide “N”, Wide “N” Knot), to the current method of suturing (MonocrylTM monofilament, discontinuous suturesmore » with a 2×2×2×2 knot) used in monitoring and research programs with a novel antiseptic barrier on the wound (“Second Skin”).« less

Lipid droplets form from distinct regions of the cell in the fission yeast Schizosaccharomyces pombe

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

Meyers, Alex; del Rio, Zuania P.; Beaver, Rachael A.

Eukaryotic cells store cholesterol/sterol esters (SEs) and triacylglycerols (TAGs) in lipid droplets, which form from the contiguous endoplasmic reticulum (ER) network. However, it is not known if droplets preferentially form from certain regions of the ER over others. Here, we used fission yeast Schizosaccharomyces pombe cells where the nuclear and cortical/peripheral ER domains are distinguishable by light microscopy to show that SE-enriched lipid droplets form away from the nucleus at the cell tips, whereas TAG-enriched lipid droplets form around the nucleus. Sterols localize to the regions of the cells where droplets enriched in SEs are observed. TAG droplet formation aroundmore » the nucleus appears to be a strong function of diacylglycerol (DAG) homeostasis with Cpt1p, which coverts DAG into phosphatidylcholine and phosphatidylethanolamine localized exclusively to the nuclear ER. Also, Dgk1p, which converts DAG into phosphatidic acid localized strongly to the nuclear ER over the cortical/peripheral ER. We also show that TAG more readily translocates from the ER to lipid droplets than do SEs. Lastly, the results augment the standard lipid droplet formation model, which has SEs and TAGs flowing into the same nascent lipid droplet regardless of its biogenesis point in the cell.« less

Lipid droplets form from distinct regions of the cell in the fission yeast Schizosaccharomyces pombe

DOE PAGES

Meyers, Alex; del Rio, Zuania P.; Beaver, Rachael A.; ...

2016-04-29

Eukaryotic cells store cholesterol/sterol esters (SEs) and triacylglycerols (TAGs) in lipid droplets, which form from the contiguous endoplasmic reticulum (ER) network. However, it is not known if droplets preferentially form from certain regions of the ER over others. Here, we used fission yeast Schizosaccharomyces pombe cells where the nuclear and cortical/peripheral ER domains are distinguishable by light microscopy to show that SE-enriched lipid droplets form away from the nucleus at the cell tips, whereas TAG-enriched lipid droplets form around the nucleus. Sterols localize to the regions of the cells where droplets enriched in SEs are observed. TAG droplet formation aroundmore » the nucleus appears to be a strong function of diacylglycerol (DAG) homeostasis with Cpt1p, which coverts DAG into phosphatidylcholine and phosphatidylethanolamine localized exclusively to the nuclear ER. Also, Dgk1p, which converts DAG into phosphatidic acid localized strongly to the nuclear ER over the cortical/peripheral ER. We also show that TAG more readily translocates from the ER to lipid droplets than do SEs. Lastly, the results augment the standard lipid droplet formation model, which has SEs and TAGs flowing into the same nascent lipid droplet regardless of its biogenesis point in the cell.« less

Effects of geolocation archival tags on reproduction and adult body mass of sooty shearwaters (Puffinus griseus)

USGS Publications Warehouse

Adams, J.; Scott, D.; McKechnie, S.; Blackwell, G.; Shaffer, S.A.; Moller, H.

2009-01-01

We attached 11 g (1.4% body-mass equivalent) global location sensing (GLS) archival tag packages to tarsi of 25 breeding sooty shearwaters (Puffinus griseus, titi) on Whenua Hou (Codfish Island), New Zealand during the chick-rearing period in 2005. Compared with chicks reared by non-handled adults that did not carry tags, deployment of tags on one or both adult parents ultimately resulted in 35% reduction in chick body mass and significantly reduced chick skeletal size preceding fledging (19 April). However, body mass between chick groups was not significantly different after controlling for skeletal size. Effects on chicks were more pronounced in six pairs where both parents carried tags. Chick mass was negatively related to the duration that adults carried tags. In this study, none of the chicks reared by pairs where both parents were tagged, 54% of chicks reared by pairs where one parent was tagged, and 83% of chicks reared by non-handled and non-tagged parents achieved a previously determined pre-fledging mass threshold (564 g; Sagar & Horning 1998). Body mass of adults carrying tags and returning from transequatorial migration the following year were 4% lighter on average than non-tagged birds, but this difference was not statistically significant. Reduced mass among chicks reared by adults carrying tags during the chick-provisioning period indicated that adults altered "normal" provisioning behaviours to maintain their own body condition at the expense of their chicks. Population-level information derived from telemetry studies can reveal important habitat-linked behaviours, unique aspects of seabird foraging behaviours, and migration ecology. Information for some species (e.g., overlap with fisheries) can aid conservation and marine ecosystem management. We advise caution, however, when interpreting certain data related to adult provisioning behaviours (e.g., time spent foraging, provisioning rates, etc.). If effects on individuals are of concern, we suggest shorter-term deployments, smaller and lighter tags, and alternative attachment techniques, especially when investigating threatened or endangered species. ?? The Royal Society of New Zealand 2009.

Notes on SAW Tag Interrogation Techniques

NASA Technical Reports Server (NTRS)

Barton, Richard J.

2010-01-01

We consider the problem of interrogating a single SAW RFID tag with a known ID and known range in the presence of multiple interfering tags under the following assumptions: (1) The RF propagation environment is well approximated as a simple delay channel with geometric power-decay constant alpha >/= 2. (2) The interfering tag IDs are unknown but well approximated as independent, identically distributed random samples from a probability distribution of tag ID waveforms with known second-order properties, and the tag of interest is drawn independently from the same distribution. (3) The ranges of the interfering tags are unknown but well approximated as independent, identically distributed realizations of a random variable rho with a known probability distribution f(sub rho) , and the tag ranges are independent of the tag ID waveforms. In particular, we model the tag waveforms as random impulse responses from a wide-sense-stationary, uncorrelated-scattering (WSSUS) fading channel with known bandwidth and scattering function. A brief discussion of the properties of such channels and the notation used to describe them in this document is given in the Appendix. Under these assumptions, we derive the expression for the output signal-to-noise ratio (SNR) for an arbitrary combination of transmitted interrogation signal and linear receiver filter. Based on this expression, we derive the optimal interrogator configuration (i.e., transmitted signal/receiver filter combination) in the two extreme noise/interference regimes, i.e., noise-limited and interference-limited, under the additional assumption that the coherence bandwidth of the tags is much smaller than the total tag bandwidth. Finally, we evaluate the performance of both optimal interrogators over a broad range of operating scenarios using both numerical simulation based on the assumed model and Monte Carlo simulation based on a small sample of measured tag waveforms. The performance evaluation results not only provide guidelines for proper interrogator design, but also provide some insight on the validity of the assumed signal model. It should be noted that the assumption that the impulse response of the tag of interest is known precisely implies that the temperature and range of the tag are also known precisely, which is generally not the case in practice. However, analyzing interrogator performance under this simplifying assumption is much more straightforward and still provides a great deal of insight into the nature of the problem.

High-throughput purification of recombinant proteins using self-cleaving intein tags.

PubMed

Coolbaugh, M J; Shakalli Tang, M J; Wood, D W

2017-01-01

High throughput methods for recombinant protein production using E. coli typically involve the use of affinity tags for simple purification of the protein of interest. One drawback of these techniques is the occasional need for tag removal before study, which can be hard to predict. In this work, we demonstrate two high throughput purification methods for untagged protein targets based on simple and cost-effective self-cleaving intein tags. Two model proteins, E. coli beta-galactosidase (βGal) and superfolder green fluorescent protein (sfGFP), were purified using self-cleaving versions of the conventional chitin-binding domain (CBD) affinity tag and the nonchromatographic elastin-like-polypeptide (ELP) precipitation tag in a 96-well filter plate format. Initial tests with shake flask cultures confirmed that the intein purification scheme could be scaled down, with >90% pure product generated in a single step using both methods. The scheme was then validated in a high throughput expression platform using 24-well plate cultures followed by purification in 96-well plates. For both tags and with both target proteins, the purified product was consistently obtained in a single-step, with low well-to-well and plate-to-plate variability. This simple method thus allows the reproducible production of highly pure untagged recombinant proteins in a convenient microtiter plate format. Copyright © 2016 Elsevier Inc. All rights reserved.

Novel Use of PIT Tags in Sea Cucumbers: Promising Results with the Commercial Species Cucumaria frondosa

PubMed Central

Gianasi, Bruno L.; Verkaik, Katie; Hamel, Jean-François; Mercier, Annie

2015-01-01

The lack of a reliable and innocuous mark-recapture method has limited studies that would provide essential information for the management of commercial sea cucumbers. Tagging sea cucumbers is notoriously difficult because of their plastic nature and autolysis capacities. The markers that have so far been tested, mainly on or through the body wall, were either lost rapidly or had major drawbacks (e.g. suitable only for batch identification, requiring complex analysis, causing infections, necrosis, behavioural changes and mortality). The present study explored the efficacy of passive integrated transponder (PIT) tags for individually marking sea cucumbers by assessing retention rates and long-term side effects of tags inserted in previously unstudied tissues/organs. Individuals of the species Cucumaria frondosa were tagged in the body wall, aquapharyngeal bulb and at the base of the oral tentacles. They were monitored closely for evidence of stress, infection, change in feeding and spawning behaviour and tag retention rate. Implanting the tag in an oral tentacle to reach the hydrovascular system of the aquapharyngeal bulb achieved the best retention rates in full-size individuals: from a maximum of 92% after 30 days to 68% at the end of the experimental period (300 days). Efficacy was lower in smaller individuals (84% after 30 d and 42% after 300 d). Following a slight increase in cloacal movements for 15 h post tagging, no side effect was noted in sea cucumbers tagged in the aquapharyngeal bulb via the tentacles. Feeding and spawning behaviours were not affected and no signs of infections or abnormal cell development in the vicinity of the tags were observed. This study indicates that marking sea cucumbers with 8.2 mm long PIT tags implanted via the oral tentacle is an effective technique, yielding relatively high retention rates over long periods without any detectable physiological or behavioural effects. PMID:26011165

Evaluation of the fish passage effectiveness of the Bonneville I prototype surface collector using three-dimensional ultrasonic fish tracking - Final Report

USGS Publications Warehouse

Faber, D.M; Weiland, M.A.; Moursund, R.A.; Carlson, T.J.; Adams, N.; Rondorf, D.

2001-01-01

This report describes tests conducted at Bonneville Dam on the Columbia River in the spring of 2000. The studies used three-dimensional (3D) acoustic telemetry and computational fluid dynamics (CFD) hydraulic modeling techniques to evaluate the response of outmigrating juvenile steelhead (Oncorhynchus mykiss) and yearling chinook (O. tshawytscha) to the Prototype Surface Collector (PSC) installed at Powerhouse I of Bonneville Dam in 1998 to test the concept of using a deep-slot surface bypass collector to divert downstream migrating salmon from turbines. The study was conducted by Pacific Northwest National Laboratory (PNNL), the Waterways Experiment Station of the U.S. Army Corp of Engineers (COE), Asci Corporation, and the U.S. Geological Survey (USGS), and was sponsored by COE’s Portland District. The goal of the study was to observe the three-dimensional behavior of tagged fish (fish bearing ultrasonic micro-transmitters) within 100 meters (m) of the surface flow bypass structure to test hypotheses about the response of migrants to flow stimuli generated by the presence of the surface flow bypass prototype and its operation. Research was done in parallel with radio telemetry studies conducted by USGS and hydroacoustic studies conducted by WES & Asci to evaluate the prototype surface collector.

Spring-summer movements of bonytail in a Colorado River reservoir, Lake Mohave, Arizona and Nevada

USGS Publications Warehouse

Marsh, Paul C.; Mueller, Gordon

1999-01-01

Bonytail can move substantial distances in a short time (10s of km in a few days). Fish in both years apparently favored the same areas, where they may remain for weeks. Unmarked bonytail were observed or captured by setting nets in places favored by tagged fish, a significant result since future use of the technique may enhance our ability to monitor reintroductions, locate and document spawning, examine habitat use, and acquire desperately needed brood stock for this critically imperiled species. External tagging techniques developed for juvenile razorback sucker may provide a method of minimizing telemetry induced stress while allowing us to focus sampling on congregation sites.

Detecting special nuclear material using muon-induced neutron emission

NASA Astrophysics Data System (ADS)

Guardincerri, Elena; Bacon, Jeffrey; Borozdin, Konstantin; Matthew Durham, J.; Fabritius, Joseph, II; Hecht, Adam; Milner, Edward C.; Miyadera, Haruo; Morris, Christopher L.; Perry, John; Poulson, Daniel

2015-07-01

The penetrating ability of cosmic ray muons makes them an attractive probe for imaging dense materials. Here, we describe experimental results from a new technique that uses neutrons generated by cosmic-ray muons to identify the presence of special nuclear material (SNM). Neutrons emitted from SNM are used to tag muon-induced fission events in actinides and laminography is used to form images of the stopping material. This technique allows the imaging of SNM-bearing objects tagged using muon tracking detectors located above or to the side of the objects, and may have potential applications in warhead verification scenarios. During the experiment described here we did not attempt to distinguish the type or grade of the SNM.

Parallel gene analysis with allele-specific padlock probes and tag microarrays

PubMed Central

Banér, Johan; Isaksson, Anders; Waldenström, Erik; Jarvius, Jonas; Landegren, Ulf; Nilsson, Mats

2003-01-01

Parallel, highly specific analysis methods are required to take advantage of the extensive information about DNA sequence variation and of expressed sequences. We present a scalable laboratory technique suitable to analyze numerous target sequences in multiplexed assays. Sets of padlock probes were applied to analyze single nucleotide variation directly in total genomic DNA or cDNA for parallel genotyping or gene expression analysis. All reacted probes were then co-amplified and identified by hybridization to a standard tag oligonucleotide array. The technique was illustrated by analyzing normal and pathogenic variation within the Wilson disease-related ATP7B gene, both at the level of DNA and RNA, using allele-specific padlock probes. PMID:12930977

American white pelican predation on Cui-ui in Pyramid Lake, Nevada

USGS Publications Warehouse

Scoppettone, Gayton G.; Rissler, Peter H.; Fabes, Mark C.; Withers, Donna

2014-01-01

Anthropogenic changes to the Pyramid Lake–Truckee River ecosystem in Nevada are suspected to have altered the predator–prey balance between American white pelican Pelecanus erythrorhynchos and Cui-ui Chasmistes cujus. We estimated the loss of the adult Cui-ui population to pelican predation over a 13-year period by netting and tagging Cui-uis as they aggregated at the mouth of the Truckee River prior to their spawning migration into the Truckee River. Cui-ui access to the Truckee River typically required traversing a shallow delta (a foraging advantage for these American white pelicans). Dams and greater frequency of low stream flows also contributed to American white pelican foraging success. We used tag recoveries from Pyramid Lake's nesting colony of American white pelicans along with an experiment to estimate the chance of tag recovery within the colony to calculate the number of tagged fish taken by American white pelicans. We also used numbered tags to test whether there was a size preference for Cui-uis taken. Our results showed that the primary source of adult Cui-ui mortality was from American white pelican predation in the Truckee River. Within a 13-year period American white pelicans had taken 90% of the tags deployed during the first 7 years of the interval. There was no preference for the size of Cui-uis taken. A better understanding of the effects of heavy cropping by American white pelicans on Cui-ui population dynamics is still needed.

Femtosecond Laser Tagging Characterization of a Sweeping Jet Actuator Operating in the Compressible Regime

NASA Technical Reports Server (NTRS)

Peters, Christopher J.; Miles, Richard B.; Burns, Ross A.; Bathel, Brett F.; Jones, Gregory S.; Danehy, Paul M.

2016-01-01

A sweeping jet (SWJ) actuator operating over a range of nozzle pressure ratios (NPRs) was characterized with femtosecond laser electronic excitation tagging (FLEET), single hot-wire anemometry (HWA) and high-speed/phase-averaged schlieren. FLEET velocimetry was successfully demonstrated in a highly unsteady, oscillatory flow containing subsonic through supersonic velocities. Qualitative comparisons between FLEET and HWA (which measured mass flux since the flow was compressible) showed relatively good agreement in the external flow profiles. The spreading rate was found to vary from 0.5 to 1.2 depending on the pressure ratio. The precision of FLEET velocity measurements in the external flow field was poorer (is approximately equal to 25 m/s) than reported in a previous study due to the use of relatively low laser fluences, impacting the velocity fluctuation measurements. FLEET enabled velocity measurements inside the device and showed that choking likely occurred for NPR = 2.0, and no internal shockwaves were present. Qualitative oxygen concentration measurements using FLEET were explored in an effort to gauge the jet's mixing with the ambient. The jet was shown to mix well within roughly four throat diameters and mix fully within roughly eight throat diameters. Schlieren provided visualization of the internal and external flow fields and showed that the qualitative structure of the internal flow does not vary with pressure ratio and the sweeping mechanism observed for incompressible NPRs also probably holds for compressible NPRs.

Population Dynamics of Adult Lost River (Deltistes luxatus) and Shortnose (Chasmistes brevirostris) Suckers in Clear Lake Reservoir, California, 2006-08

USGS Publications Warehouse

Barry, Patrick M.; Janney, Eric C.; Hewitt, David A.; Hayes, Brian S.; Scott, Alta C.

2009-01-01

We report results from ongoing research into the population dynamics of endangered Lost River and shortnose suckers in Clear Lake Reservoir, California. Results are included for sampling that occurred from fall 2006 to spring 2008. We summarize catches and passive integrated transponder tagging efforts from trammel net sampling in fall 2006 and fall 2007, and report on detections of tagged suckers on remote antennas in the primary spawning tributary, Willow Creek, in spring 2007 and spring 2008. Results from trammel net sampling were similar to previous years, although catches of suckers in fall 2006 were lower than in 2007 and past years. Lost River and shortnose suckers combined made up about 80 percent of the sucker catch in each year, and more than 2,000 new fish were tagged across the 2 years. Only a small number of the suckers captured in fall sampling were recaptures of previously tagged fish, reinforcing the importance of remote detections of fish for capture-recapture analysis. Detections of tagged suckers in Willow Creek were low in spring 2007, presumably because of low flows. Nonetheless, the proportions of tagged fish that were detected were reasonably high and capture-recapture analyses should be possible after another year of data collection. Run timing for Lost River and shortnose suckers was well described by first detections of individuals by antennas in Willow Creek, although we may not have installed the antennas early enough in 2008 to monitor the earliest portion of the Lost River sucker migration. The duration and magnitude of the spawning runs for both species were influenced by flows and water temperature. Flows in Willow Creek were much higher in 2008 than in 2007, and far more detections were recorded in 2008 and the migrations were more protracted. In both years and for both species, migrations began in early March at water temperatures between 5 and 6 deg C and peaks were related to periods of increasing water temperature. The sex ratio of Lost River suckers detected in Willow Creek was skewed toward males, despite consistently more females having been tagged in fall sampling. This pattern indicates that some tagged female Lost River suckers may be spawning elsewhere in the system, and we intend to investigate this possibility to verify or alter the representativeness of our spring monitoring. Length frequency analysis of fall trammel net catches showed that the populations of both species in Clear Lake Reservoir have undergone major demographic transitions during the last 15 years. In the mid-1990s, the populations were dominated by larger fish and showed little evidence of recent recruitment. These larger fish apparently disappeared in the late 1990s and early 2000s, and the populations are now dominated by fish that recruited into the adult populations in the late 1990s. The length frequencies from the last 4 years provide evidence of consistent recruitment into the Lost River sucker population, but provide no such evidence for the shortnose sucker population. Overall, annual growth rates for both species in Clear Lake were 2-4 times greater than growth rates for conspecifics in Upper Klamath Lake. However, little or no growth occurred for either species in Clear Lake between 2006 and 2007. Based on available evidence, we are unable to fully explain differences in growth rates between systems or among years within Clear Lake.

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

Rowe, Nathan C; Younkin, James R; Pickett, Chris A

Radio-Frequency (RF) devices have revolutionized many aspects of modern industrial processes. RF technology can enable wireless communication for tag identification, sensor communication, and asset tracking. Radio-frequency identification (RFID) is a technology that utilizes wireless communication to interrogate and identify an electronic tag attached to an item in order to identify the item. The technology can come in many forms: passive or active tags, low to ultra-wideband frequencies, small paper-thin tags to brick-sized units, and simple tags or highly integrated sensor packages. RF technology, and specifically RFID, has been applied widely in commercial markets for inventory, supply chain management, and assetmore » tracking. Several recent studies have demonstrated the safeguards benefits of utilizing RFID versus conventional inventory tagging methods for tracking nuclear material. These studies have indicated that the RF requirements for safeguards functions are more stringent than the RF requirements for other inventory tracking and accounting applications. Additionally, other requirements must be addressed, including environmental and operating conditions, authentication, and tag location and attachment. Facility restrictions on radio spectrum, method of tag attachment, and sensitivity of the data collected impact the tag selection and system design. More important, the intended use of the system must be considered. The requirements for using RF to simply replace or supplement container identifiers such as bar codes that facilitate the inventory function will differ greatly from the requirements for deploying RF for unattended monitoring applications. Several studies have investigated these considerations to advance commercial RF devices for safeguards use, and a number of system concepts have been developed. This paper will provide an overview of past studies and current technologies, and will investigate the requirements, existing gaps, and several potential next steps for advancing RF techniques for safeguards use.« less

A review of radio frequency identification technology for the anatomic pathology or biorepository laboratory: Much promise, some progress, and more work needed.

PubMed

Lou, Jerry J; Andrechak, Gary; Riben, Michael; Yong, William H

2011-01-01

Patient safety initiatives throughout the anatomic laboratory and in biorepository laboratories have mandated increasing emphasis on the need for accurately identifying and tracking biospecimen assets throughout their production lifecycle and for archiving/retrieval purposes. However, increasing production volume along with complex workflow characteristics, reliance on manual production processes, and required asset movement to disparate destinations throughout asset lifecycles continue to challenge laboratory efforts. Radio Frequency Identification (RFID) technology, use of radio waves to communicate data between electronic tags attached to objects and a reader, shows significant potential to facilitate and overcome these hurdles. Advantages over traditional barcode labeling include readability without direct line-of-sight alignment to the reader, ability to read multiple tags simultaneously, higher data storage capacity, faster data transmission rate, and capacity to perform multiple read-writes of data to the tag. Most importantly, use of radio waves decreases the need to manually scan each asset, and at each step, identification or tracking event is needed. Temperature monitoring by on-board sensors and three-dimensional position tracking are additional potential benefits of using RFID technology. To date, barriers to implementation of RFID systems in the anatomic laboratory include increased associated costs of tags and readers, system software, data security concerns, lack of specific data standards for stored information, and potential for technological obsolescence during decades of specimen storage. Novel RFID production techniques and increased production capacity are projected to lower costs of some tags to a few cents each. Potentially, information security concerns can be addressed by techniques such as shielding, data encryption, and tag pseudonyms. Commitment by stakeholder groups to develop RFID tag data standards for anatomic pathology and biorepository laboratories could avoid or mitigate the "islands of data" dilemma presented by barcode usage where there are innumerable standards and a consequent paucity of hardware or software "plug and play" interoperability. Work remains to be done to establish the durability and appropriate shielding of individual tag types for use in harsh laboratory environmental conditions, and for long-term archival storage. Finally, given the requirements for long-term storage of biospecimen assets, consideration should be given to ways of mitigating data isolation due to eventual technological obsolescence of a particular RFID technology or software.

A review of radio frequency identification technology for the anatomic pathology or biorepository laboratory: Much promise, some progress, and more work needed

PubMed Central

Lou, Jerry J.; Andrechak, Gary; Riben, Michael; Yong, William H.

2011-01-01

Patient safety initiatives throughout the anatomic laboratory and in biorepository laboratories have mandated increasing emphasis on the need for accurately identifying and tracking biospecimen assets throughout their production lifecycle and for archiving/retrieval purposes. However, increasing production volume along with complex workflow characteristics, reliance on manual production processes, and required asset movement to disparate destinations throughout asset lifecycles continue to challenge laboratory efforts. Radio Frequency Identification (RFID) technology, use of radio waves to communicate data between electronic tags attached to objects and a reader, shows significant potential to facilitate and overcome these hurdles. Advantages over traditional barcode labeling include readability without direct line-of-sight alignment to the reader, ability to read multiple tags simultaneously, higher data storage capacity, faster data transmission rate, and capacity to perform multiple read-writes of data to the tag. Most importantly, use of radio waves decreases the need to manually scan each asset, and at each step, identification or tracking event is needed. Temperature monitoring by on-board sensors and three-dimensional position tracking are additional potential benefits of using RFID technology. To date, barriers to implementation of RFID systems in the anatomic laboratory include increased associated costs of tags and readers, system software, data security concerns, lack of specific data standards for stored information, and potential for technological obsolescence during decades of specimen storage. Novel RFID production techniques and increased production capacity are projected to lower costs of some tags to a few cents each. Potentially, information security concerns can be addressed by techniques such as shielding, data encryption, and tag pseudonyms. Commitment by stakeholder groups to develop RFID tag data standards for anatomic pathology and biorepository laboratories could avoid or mitigate the “islands of data” dilemma presented by barcode usage where there are innumerable standards and a consequent paucity of hardware or software “plug and play” interoperability. Work remains to be done to establish the durability and appropriate shielding of individual tag types for use in harsh laboratory environmental conditions, and for long-term archival storage. Finally, given the requirements for long-term storage of biospecimen assets, consideration should be given to ways of mitigating data isolation due to eventual technological obsolescence of a particular RFID technology or software. PMID:21886890

Sediment motion and velocity in a glacier-fed stream

NASA Astrophysics Data System (ADS)

Mao, L.; Dell'Agnese, A.; Comiti, F.

2017-08-01

Current understanding of coarse sediment transport (e.g. threshold for motion, travel length and virtual velocity) in mountain rivers is still quite limited, and even less is known about glacial streams. However, the hydrological characteristics of these systems (strong daily discharge fluctuations, high water turbidity) pose challenges to the use of tracers to monitor bed sediment dynamics, as tagged clasts are usually located after bedload events when flow stage has receded, e.g. by means of portable antennas in the case of Passive Integrated Transponders (PIT). The use of stationary antennas, still scarcely in use worldwide, to detect PIT-tagged particles has potential advantages in glacier-fed streams. If water discharge is monitored continuously, a stationary antenna provides real time data on the actual discharge at the moment of tracer particles passage. This study focuses on incipient motion and virtual velocity of bed particles measured by a stationary antennas system in a steep mountain channel (Saldur River, drainage area 18.6 km2, Italian Alps) where significant daily discharge fluctuations and bedload transport occur as a result of a nivo-glacial regime. Four stationary antennas were installed 50-m apart in the study reach. A total of 629 PIT-tagged clasts were inserted in the studied reach between 2011 and 2014, ranging in size from 35 mm to 580 mm, with an overall recovery rate of around 44%. Critical discharge for sediment entrainment was obtained by detecting the movement of tracers placed immediately upstream of antennas. Virtual velocity was derived by knowing distances between the antennas and travel time of tracers. Results on initiation of motion show that the relationship between the size of transported tracers and the discharge measured at the time clasts were passing the stationary antenna is very weak. The influence of antecedent flows on incipient motion was thus investigated by dividing the highest discharge recorded between each PIT deployment and the subsequent entrainment by the actual critical discharge at the time of movement (ratio Qmax/Qc). Results show that approximately 50% of tracers moved at Qmax/Qc ≤ 1.2, and that 73% of tracers moved at Qmax/Qc < 1.5. Therefore, about 30% of tracers had to previously experience a discharge substantially greater than the one that actually mobilized them. Also, coarser particles moved at higher Qmax/Qc ratios, suggesting that higher antecedent flows may be needed to destabilize bed clustering. Results on the virtual velocity of the PIT-tagged clasts employed in the field show that the virtual velocity turned out to be highly variable (ranging from 101 to 10- 5 m min- 1) and weakly related to either particle size or flow discharge. However, virtual velocity was well correlated with the highest flow discharge experienced by each tracer normalized by a percentile of the flow duration curve. This evidence further stresses the importance of flow history on sediment entrainment and transport. Finally, the pros and cons of the deployed monitoring technology are discussed.

Targeting Breast Cancer Vasculature

DTIC Science & Technology

2006-03-01

E., and Hanahan, D. Stage-specific vascular markers revealed by phage display in a mouse model of pancreatic islet tumorigenesis. Cancer Cell 4:393...expressing full-length myc-tagged metadherin, myc-vimen- tin, or myc-pCMV were analyzed by flow cytometry . Anti-myc antibodies were applied to the cells...In 4T1 tumor cell extracts, anti-metadherin(378-440) detectedthen stained with anti-myc antibodies. Using flow cytometry , proteins with apparent

Assessing potential errors of MRI-based measurements of pulmonary blood flow using a detailed network flow model

PubMed Central

Buxton, R. B.; Prisk, G. K.

2012-01-01

MRI images of pulmonary blood flow using arterial spin labeling (ASL) measure the delivery of magnetically tagged blood to an image plane during one systolic ejection period. However, the method potentially suffers from two problems, each of which may depend on the imaging plane location: 1) the inversion plane is thicker than the imaging plane, resulting in a gap that blood must cross to be detected in the image; and 2) ASL includes signal contributions from tagged blood in conduit vessels (arterial and venous). By using an in silico model of the pulmonary circulation we found the gap reduced the ASL signal to 64–74% of that in the absence of a gap in the sagittal plane and 53–84% in the coronal. The contribution of the conduit vessels varied markedly as a function of image plane ranging from ∼90% of the overall signal in image planes that encompass the central hilar vessels to <20% in peripheral image planes. A threshold cutoff removing voxels with intensities >35% of maximum reduced the conduit vessel contribution to the total ASL signal to ∼20% on average; however, planes with large contributions from conduit vessels underestimate acinar flow due to a high proportion of in-plane flow, making ASL measurements of perfusion impractical. In other image planes, perfusion dominated the resulting ASL images with good agreement between ASL and acinar flow. Similarly, heterogeneity of the ASL signal as measured by relative dispersion is a reliable measure of heterogeneity of the acinar flow distribution in the same image planes. PMID:22539167

Assessing potential errors of MRI-based measurements of pulmonary blood flow using a detailed network flow model.

PubMed

Burrowes, K S; Buxton, R B; Prisk, G K

2012-07-01

MRI images of pulmonary blood flow using arterial spin labeling (ASL) measure the delivery of magnetically tagged blood to an image plane during one systolic ejection period. However, the method potentially suffers from two problems, each of which may depend on the imaging plane location: 1) the inversion plane is thicker than the imaging plane, resulting in a gap that blood must cross to be detected in the image; and 2) ASL includes signal contributions from tagged blood in conduit vessels (arterial and venous). By using an in silico model of the pulmonary circulation we found the gap reduced the ASL signal to 64-74% of that in the absence of a gap in the sagittal plane and 53-84% in the coronal. The contribution of the conduit vessels varied markedly as a function of image plane ranging from ∼90% of the overall signal in image planes that encompass the central hilar vessels to <20% in peripheral image planes. A threshold cutoff removing voxels with intensities >35% of maximum reduced the conduit vessel contribution to the total ASL signal to ∼20% on average; however, planes with large contributions from conduit vessels underestimate acinar flow due to a high proportion of in-plane flow, making ASL measurements of perfusion impractical. In other image planes, perfusion dominated the resulting ASL images with good agreement between ASL and acinar flow. Similarly, heterogeneity of the ASL signal as measured by relative dispersion is a reliable measure of heterogeneity of the acinar flow distribution in the same image planes.

Population-scale movement of coastal cutthroat trout in a naturally isolated stream network

USGS Publications Warehouse

Gresswell, R.E.; Hendricks, S.R.

2007-01-01

To identify population-scale patterns of movement, coastal cutthroat trout Oncorhynchus clarkii clarkii tagged and marked (35 radio-tagged, 749 passive integrated transponder [PIT]-tagged, and 3,025 fin-clipped) were monitored from June 1999 to August 2000. The study watershed, located in western Oregon, was above a natural barrier to upstream movement. Emigration out of the watershed was estimated with a rotating fish trap. Approximately 70% of recaptured coastal cutthroat trout with PIT tags and 86% of those with radio tags moved predominantly at the channel-unit scale (2-95 m); fewer tagged fish moved at the reach scale (66-734 m) and segment scale (229-3,479 m). In general, movement was greatest in April as spawning peaked and lowest in October, when discharge was at its lowest. Only 63 (<1% of tagged and marked fish) coastal cutthroat trout were captured in the fish trap. Trap efficiency was about 33%, and the expanded estimate of emigrants between February and June was 173 fish. These results suggest that unit-scale movement is common throughout the year and that reach- and segment-scale movements are important during the winter and spring. Although movement in headwater streams is most common at the channel-unit scale, restoration of individual channel units of stream may not benefit the population at the watershed scale unless these activities are undertaken in the context of the greater whole. Individual coastal cutthroat trout move great distances, even within the small watersheds in the Oregon Coast Range, and although these movements may be infrequent, they may contribute substantially to recolonization after stochastic extirpation events (e.g., landslides and debris flows). Management strategies that focus on maintaining and restoring connectivity in a watershed represent an important step toward protecting the evolutionary capacity of stream salmonids. ??

Contrasting Life Strategies of Viruses that Infect Photo- and Heterotrophic Bacteria, as Revealed by Viral Tagging

PubMed Central

Deng, Li; Gregory, Ann; Yilmaz, Suzan; Poulos, Bonnie T.; Hugenholtz, Philip; Sullivan, Matthew B.

2012-01-01

ABSTRACT Ocean viruses are ubiquitous and abundant and play important roles in global biogeochemical cycles by means of their mortality, horizontal gene transfer, and manipulation of host metabolism. However, the obstacles involved in linking viruses to their hosts in a high-throughput manner bottlenecks our ability to understand virus-host interactions in complex communities. We have developed a method called viral tagging (VT), which combines mixtures of host cells and fluorescent viruses with flow cytometry. We investigated multiple viruses which infect each of two model marine bacteria that represent the slow-growing, photoautotrophic genus Synechococcus (Cyanobacteria) and the fast-growing, heterotrophic genus Pseudoalteromonas (Gammaproteobacteria). Overall, viral tagging results for viral infection were consistent with plaque and liquid infection assays for cyanobacterial myo-, podo- and siphoviruses and some (myo- and podoviruses) but not all (four siphoviruses) heterotrophic bacterial viruses. Virus-tagged Pseudoalteromonas organisms were proportional to the added viruses under varied infection conditions (virus-bacterium ratios), while no more than 50% of the Synechococcus organisms were virus tagged even at viral abundances that exceeded (5 to 10×) that of their hosts. Further, we found that host growth phase minimally impacts the fraction of virus-tagged Synechococcus organisms while greatly affecting phage adsorption to Pseudoalteromonas. Together these findings suggest that at least two contrasting viral life strategies exist in the oceans and that they likely reflect adaptation to their host microbes. Looking forward to the point at which the virus-tagging signature is well understood (e.g., for Synechococcus), application to natural communities should begin to provide population genomic data at the proper scale for predictively modeling two of the most abundant biological entities on Earth. PMID:23111870

Non-Contrast-Enhanced Renal Angiography Using Multiple Inversion Recovery and Alternating TR Balanced Steady State Free Precession

PubMed Central

Dong, Hattie Z.; Worters, Pauline W.; Wu, Holden H.; Ingle, R. Reeve; Vasanawala, Shreyas S.; Nishimura, Dwight G.

2014-01-01

Non-contrast enhanced renal angiography techniques based on balanced steady state free precession (SSFP) avoid external contrast agents, take advantage of high inherent blood signal from the T2/T1 contrast mechanism, and have short SSFP acquisition times. However, background suppression is limited; inflow times are inflexible; labeling region is difficult to define when tagging arterial flow; and scan times are long. To overcome these limitations, we propose the use of multiple inversion recovery (MIR) preparatory pulses combined with alternating TR balanced SSFP (ATR-SSFP) to produce renal angiograms. MIR uses selective spatial saturation followed by four global inversion recovery pulses to concurrently null a wide range of background T1 species while allowing for adjustable inflow times; ATR-SSFP maintains vessel contrast and provides added fat suppression. The high level of suppression enables imaging in 3D as well as projective 2D formats, the latter of which has a scan time down to one heartbeat. In vivo studies at 1.5 T demonstrate the superior vessel contrast of this technique. PMID:23172805

Live single-cell laser tag.

PubMed

Binan, Loïc; Mazzaferri, Javier; Choquet, Karine; Lorenzo, Louis-Etienne; Wang, Yu Chang; Affar, El Bachir; De Koninck, Yves; Ragoussis, Jiannis; Kleinman, Claudia L; Costantino, Santiago

2016-05-20

The ability to conduct image-based, non-invasive cell tagging, independent of genetic engineering, is key to cell biology applications. Here we introduce cell labelling via photobleaching (CLaP), a method that enables instant, specific tagging of individual cells based on a wide array of criteria such as shape, behaviour or positional information. CLaP uses laser illumination to crosslink biotin onto the plasma membrane, coupled with streptavidin conjugates to label individual cells for genomic, cell-tracking, flow cytometry or ultra-microscopy applications. We show that the incorporated mark is stable, non-toxic, retained for several days, and transferred by cell division but not to adjacent cells in culture. To demonstrate the potential of CLaP for genomic applications, we combine CLaP with microfluidics-based single-cell capture followed by transcriptome-wide next-generation sequencing. Finally, we show that CLaP can also be exploited for inducing transient cell adhesion to substrates for microengineering cultures with spatially patterned cell types.

Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding.

PubMed

Shahi, Payam; Kim, Samuel C; Haliburton, John R; Gartner, Zev J; Abate, Adam R

2017-03-14

Proteins are the primary effectors of cellular function, including cellular metabolism, structural dynamics, and information processing. However, quantitative characterization of proteins at the single-cell level is challenging due to the tiny amount of protein available. Here, we present Abseq, a method to detect and quantitate proteins in single cells at ultrahigh throughput. Like flow and mass cytometry, Abseq uses specific antibodies to detect epitopes of interest; however, unlike these methods, antibodies are labeled with sequence tags that can be read out with microfluidic barcoding and DNA sequencing. We demonstrate this novel approach by characterizing surface proteins of different cell types at the single-cell level and distinguishing between the cells by their protein expression profiles. DNA-tagged antibodies provide multiple advantages for profiling proteins in single cells, including the ability to amplify low-abundance tags to make them detectable with sequencing, to use molecular indices for quantitative results, and essentially limitless multiplexing.

Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding

NASA Astrophysics Data System (ADS)

Shahi, Payam; Kim, Samuel C.; Haliburton, John R.; Gartner, Zev J.; Abate, Adam R.

2017-03-01

Proteins are the primary effectors of cellular function, including cellular metabolism, structural dynamics, and information processing. However, quantitative characterization of proteins at the single-cell level is challenging due to the tiny amount of protein available. Here, we present Abseq, a method to detect and quantitate proteins in single cells at ultrahigh throughput. Like flow and mass cytometry, Abseq uses specific antibodies to detect epitopes of interest; however, unlike these methods, antibodies are labeled with sequence tags that can be read out with microfluidic barcoding and DNA sequencing. We demonstrate this novel approach by characterizing surface proteins of different cell types at the single-cell level and distinguishing between the cells by their protein expression profiles. DNA-tagged antibodies provide multiple advantages for profiling proteins in single cells, including the ability to amplify low-abundance tags to make them detectable with sequencing, to use molecular indices for quantitative results, and essentially limitless multiplexing.

Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding

PubMed Central

Shahi, Payam; Kim, Samuel C.; Haliburton, John R.; Gartner, Zev J.; Abate, Adam R.

2017-01-01

Proteins are the primary effectors of cellular function, including cellular metabolism, structural dynamics, and information processing. However, quantitative characterization of proteins at the single-cell level is challenging due to the tiny amount of protein available. Here, we present Abseq, a method to detect and quantitate proteins in single cells at ultrahigh throughput. Like flow and mass cytometry, Abseq uses specific antibodies to detect epitopes of interest; however, unlike these methods, antibodies are labeled with sequence tags that can be read out with microfluidic barcoding and DNA sequencing. We demonstrate this novel approach by characterizing surface proteins of different cell types at the single-cell level and distinguishing between the cells by their protein expression profiles. DNA-tagged antibodies provide multiple advantages for profiling proteins in single cells, including the ability to amplify low-abundance tags to make them detectable with sequencing, to use molecular indices for quantitative results, and essentially limitless multiplexing. PMID:28290550

Three-dimensional seismic structure of a Mid-Atlantic Ridge segment characterized by active detachment faulting (TAG, 25°55’N-26°20’N)

NASA Astrophysics Data System (ADS)

Zhao, M.; Canales, J.

2009-12-01

The Trans-Atlantic Geotraverse (TAG) segment of the Mid-Atlantic Ridge (MAR) (25°55'N-26°20'N) is characterized by massive active and relict high-temperature hydrothermal deposits. Previous geological and geophysical studies indicate that the active TAG hydrothermal mound sits on the hanging wall of an active detachment fault. The STAG microseismicity study revealed that seismicity associated to detachment faulting extends deep into the crust/uppermost mantle (>6 km), forming an arcuate band (in plan view) extending along ~25 km of the rift valley floor (deMartin et al., Geology, 35, 711-714, 2007). Two-dimensional analysis of the STAG seismic refraction data acquired with ocean bottom seismometers (OBSs) showed that the eastern rift valley wall is associated with high P-wave velocities (>7 km/s) at shallow levels (>1 km depth), indicating uplift of lower crustal and/or upper mantle rocks along the detachment fault (Canales et al., Geochem., Geophys., Geosyst., 8, Q08004, doi:08010.01029/02007GC001629, 2008). Here we present a three-dimensional (3D) seismic tomography analysis of the complete STAG seismic refraction OBS dataset to illuminate the 3D crustal architecture of the TAG segment. Our new results provide, for the first time, a detailed picture of the complex, dome-shaped geometry and structure of a nascent oceanic core complex being exhumed by a detachment fault. Our results show a relatively low-velocity anomaly embedded within the high-velocity body forming the footwall of the detachment fault. The low velocity sits 2-3 km immediately beneath the active TAG hydrothermal mound. Although velocities within the low-velocity zone are too high (6 km/s) to represent partial melt, we speculate that this low velocity zone is intimately linked to hydrothermal processes taking place at TAG. We consider three possible scenarios for its origin: (1) a highly fissured zone produced by extensional stresses during footwall exhumation that may help localize fluid flow; (2) a hot -perhaps partially molten- gabbro pluton intruding the detachment fault footwall, which could provide some of the heat driving hydrothermal circulation at TAG; or (3) serpenitized peridotite, with hydration of the footwall being enhanced by hydrothermal fluid flow. This research was granted by the US-NSF (OCE-0137329) and the Chinese National Natural Science Foundation (40776025). M. Zhao was supported by China Scholarship Council (CSC) for 6 months of cooperative research at WHOI.

Difficulties in Generating Specific Antibodies for Immunohistochemical Detection of Nitrosylated Tubulins

PubMed Central

Kamnev, Anton; Muhar, Matthias; Preinreich, Martina; Ammer, Hermann; Propst, Friedrich

2013-01-01

Protein S-nitrosylation, the covalent attachment of a nitroso moiety to thiol groups of specific cysteine residues, is one of the major pathways of nitric oxide signaling. Hundreds of proteins are subject to this transient post-translational modification and for some the functional consequences have been identified. Biochemical assays for the analysis of protein S-nitrosylation have been established and can be used to study if and under what conditions a given protein is S-nitrosylated. In contrast, the equally desirable subcellular localization of specific S-nitrosylated protein isoforms has not been achieved to date. In the current study we attempted to specifically localize S-nitrosylated α- and β-tubulin isoforms in primary neurons after fixation. The approach was based on in situ replacement of the labile cysteine nitroso modification with a stable tag and the subsequent use of antibodies which recognize the tag in the context of the tubulin polypeptide sequence flanking the cysteine residue of interest. We established a procedure for tagging S-nitrosylated proteins in cultured primary neurons and obtained polyclonal anti-tag antibodies capable of specifically detecting tagged proteins on immunoblots and in fixed cells. However, the antibodies were not specific for tubulin isoforms. We suggest that different tagging strategies or alternative methods such as fluorescence resonance energy transfer techniques might be more successful. PMID:23840827

Evaluation of Intercontinental Transport of Ozone Using Full-tagged, Tagged-N and Sensitivity Methods

NASA Astrophysics Data System (ADS)

Guo, Y.; Liu, J.; Mauzerall, D. L.; Emmons, L. K.; Horowitz, L. W.; Fan, S.; Li, X.; Tao, S.

2014-12-01

Long-range transport of ozone is of great concern, yet the source-receptor relationships derived previously depend strongly on the source attribution techniques used. Here we describe a new tagged ozone mechanism (full-tagged), the design of which seeks to take into account the combined effects of emissions of ozone precursors, CO, NOx and VOCs, from a particular source, while keeping the current state of chemical equilibrium unchanged. We label emissions from the target source (A) and background (B). When two species from A and B sources react with each other, half of the resulting products are labeled A, and half B. Thus the impact of a given source on downwind regions is recorded through tagged chemistry. We then incorporate this mechanism into the Model for Ozone and Related chemical Tracers (MOZART-4) to examine the impact of anthropogenic emissions within North America, Europe, East Asia and South Asia on ground-level ozone downwind of source regions during 1999-2000. We compare our results with two previously used methods -- the sensitivity and tagged-N approaches. The ozone attributed to a given source by the full-tagged method is more widely distributed spatially, but has weaker seasonal variability than that estimated by the other methods. On a seasonal basis, for most source/receptor pairs, the full-tagged method estimates the largest amount of tagged ozone, followed by the sensitivity and tagged-N methods. In terms of trans-Pacific influence of ozone pollution, the full-tagged method estimates the strongest impact of East Asian (EA) emissions on the western U.S. (WUS) in MAM and JJA (~3 ppbv), which is substantially different in magnitude and seasonality from tagged-N and sensitivity studies. This difference results from the full-tagged method accounting for the maintenance of peroxy radicals (e.g., CH3O2, CH3CO3, and HO2), in addition to NOy, as effective reservoirs of EA source impact across the Pacific, allowing for a significant contribution to ozone formation over WUS (particularly in summer). Thus, the full-tagged method, with its clear discrimination of source and background contributions on a per-reaction basis, provides unique insights into the critical role of VOCs (and additional reactive nitrogen species) in determining the nonlinear inter-continental influence of ozone pollution.

A Herbivore Tag-and-Trace System Reveals Contact- and Density-Dependent Repellence of a Root Toxin.

PubMed

Bont, Zoe; Arce, Carla; Huber, Meret; Huang, Wei; Mestrot, Adrien; Sturrock, Craig J; Erb, Matthias

2017-03-01

Foraging behavior of root feeding organisms strongly affects plant-environment-interactions and ecosystem processes. However, the impact of plant chemistry on root herbivore movement in the soil is poorly understood. Here, we apply a simple technique to trace the movement of soil-dwelling insects in their habitats without disturbing or restricting their interactions with host plants. We tagged the root feeding larvae of Melolontha melolontha with a copper ring and repeatedly located their position in relation to their preferred host plant, Taraxacum officinale, using a commercial metal detector. This method was validated and used to study the influence of the sesquiterpene lactone taraxinic acid β-D-glucopyranosyl ester (TA-G) on the foraging of M. melolontha. TA-G is stored in the latex of T. officinale and protects the roots from herbivory. Using behavioral arenas with TA-G deficient and control plants, we tested the impact of physical root access and plant distance on the effect of TA-G on M. melolontha. The larvae preferred TA-G deficient plants to control plants, but only when physical root contact was possible and the plants were separated by 5 cm. Melolontha melolontha showed no preference for TA-G deficient plants when the plants were grown 15 cm apart, which may indicate a trade-off between the cost of movement and the benefit of consuming less toxic food. We demonstrate that M. melolontha integrates host plant quality and distance into its foraging patterns and suggest that plant chemistry affects root herbivore behavior in a plant-density dependent manner.

Fluorescence Imaging Reveals Surface Contamination

NASA Technical Reports Server (NTRS)

Schirato, Richard; Polichar, Raulf

1992-01-01

In technique to detect surface contamination, object inspected illuminated by ultraviolet light to make contaminants fluoresce; low-light-level video camera views fluorescence. Image-processing techniques quantify distribution of contaminants. If fluorescence of material expected to contaminate surface is not intense, tagged with low concentration of dye.

Fluorescence lifetime endoscopy using TCSPC for the measurement of FRET in live cells

PubMed Central

Fruhwirth, Gilbert O.; Ameer-Beg, Simon; Cook, Richard; Watson, Timothy; Ng, Tony; Festy, Frederic

2010-01-01

Development of remote imaging for diagnostic purposes has progressed dramatically since endoscopy began in the 1960’s. The recent advent of a clinically licensed intensity-based fluorescence micro-endoscopic instrument has offered the prospect of real-time cellular resolution imaging. However, interrogating protein-protein interactions deep inside living tissue requires precise fluorescence lifetime measurements to derive the Förster resonance energy transfer between two tagged fluorescent markers. We developed a new instrument combining remote fiber endoscopic cellular-resolution imaging with TCSPC-FLIM technology to interrogate and discriminate mixed fluorochrome labeled beads and expressible GFP/TagRFP tags within live cells. Endoscopic-FLIM (e-FLIM) data was validated by comparison with data acquired via conventional FLIM and e-FLIM was found to be accurate for both bright bead and dim live cell samples. The fiber based micro-endoscope allowed remote imaging of 4 µm and 10 µm beads within a thick Matrigel matrix with confident fluorophore discrimination using lifetime information. More importantly, this new technique enabled us to reliably measure protein-protein interactions in live cells embedded in a 3D matrix, as demonstrated by the dimerization of the fluorescent protein-tagged membrane receptor CXCR4. This cell-based application successfully demonstrated the suitability and great potential of this new technique for in vivo pre-clinical biomedical and possibly human clinical applications. PMID:20588974

Profiling cellular protein complexes by proximity ligation with dual tag microarray readout.

PubMed

Hammond, Maria; Nong, Rachel Yuan; Ericsson, Olle; Pardali, Katerina; Landegren, Ulf

2012-01-01

Patterns of protein interactions provide important insights in basic biology, and their analysis plays an increasing role in drug development and diagnostics of disease. We have established a scalable technique to compare two biological samples for the levels of all pairwise interactions among a set of targeted protein molecules. The technique is a combination of the proximity ligation assay with readout via dual tag microarrays. In the proximity ligation assay protein identities are encoded as DNA sequences by attaching DNA oligonucleotides to antibodies directed against the proteins of interest. Upon binding by pairs of antibodies to proteins present in the same molecular complexes, ligation reactions give rise to reporter DNA molecules that contain the combined sequence information from the two DNA strands. The ligation reactions also serve to incorporate a sample barcode in the reporter molecules to allow for direct comparison between pairs of samples. The samples are evaluated using a dual tag microarray where information is decoded, revealing which pairs of tags that have become joined. As a proof-of-concept we demonstrate that this approach can be used to detect a set of five proteins and their pairwise interactions both in cellular lysates and in fixed tissue culture cells. This paper provides a general strategy to analyze the extent of any pairwise interactions in large sets of molecules by decoding reporter DNA strands that identify the interacting molecules.

Interactive vs. automatic ultrasound image segmentation methods for staging hepatic lipidosis.

PubMed

Weijers, Gert; Starke, Alexander; Haudum, Alois; Thijssen, Johan M; Rehage, Jürgen; De Korte, Chris L

2010-07-01

The aim of this study was to test the hypothesis that automatic segmentation of vessels in ultrasound (US) images can produce similar or better results in grading fatty livers than interactive segmentation. A study was performed in postpartum dairy cows (N=151), as an animal model of human fatty liver disease, to test this hypothesis. Five transcutaneous and five intraoperative US liver images were acquired in each animal and a liverbiopsy was taken. In liver tissue samples, triacylglycerol (TAG) was measured by biochemical analysis and hepatic diseases other than hepatic lipidosis were excluded by histopathologic examination. Ultrasonic tissue characterization (UTC) parameters--Mean echo level, standard deviation (SD) of echo level, signal-to-noise ratio (SNR), residual attenuation coefficient (ResAtt) and axial and lateral speckle size--were derived using a computer-aided US (CAUS) protocol and software package. First, the liver tissue was interactively segmented by two observers. With increasing fat content, fewer hepatic vessels were visible in the ultrasound images and, therefore, a smaller proportion of the liver needed to be excluded from these images. Automatic-segmentation algorithms were implemented and it was investigated whether better results could be achieved than with the subjective and time-consuming interactive-segmentation procedure. The automatic-segmentation algorithms were based on both fixed and adaptive thresholding techniques in combination with a 'speckle'-shaped moving-window exclusion technique. All data were analyzed with and without postprocessing as contained in CAUS and with different automated-segmentation techniques. This enabled us to study the effect of the applied postprocessing steps on single and multiple linear regressions ofthe various UTC parameters with TAG. Improved correlations for all US parameters were found by using automatic-segmentation techniques. Stepwise multiple linear-regression formulas where derived and used to predict TAG level in the liver. Receiver-operating-characteristics (ROC) analysis was applied to assess the performance and area under the curve (AUC) of predicting TAG and to compare the sensitivity and specificity of the methods. Best speckle-size estimates and overall performance (R2 = 0.71, AUC = 0.94) were achieved by using an SNR-based adaptive automatic-segmentation method (used TAG threshold: 50 mg/g liver wet weight). Automatic segmentation is thus feasible and profitable.

Fluid pipeline leak detection and location with miniature RF tags

DOEpatents

McIntyre, Timothy J.

2017-05-16

Sensors locate troublesome leaks in pipes or conduits that carry a flowing medium. These sensors, through tailored physical and geometric properties, preferentially seek conduit leaks or breaches due to flow streaming. The sensors can be queried via transceivers outside the conduit or located and interrogated inside by submersible unmanned vehicle to identify and characterize the nature of a leak. The sensors can be functionalized with other capabilities for additional leak and pipeline characterization if needed. Sensors can be recovered from a conduit flow stream and reused for future leak detection activities.

Hydroxyl Tagging Velocimetry in Cavity-Piloted Mach 2 Combustor (Postprint)

DTIC Science & Technology

2006-01-01

combustor with a wall cavity flameholder. In the HTV method, ArF excimer laser (193 nm) beams pass through a humid gas flow and dissociate H2O into H...grid of OH tracked by planar laser -induced fluorescence to yield about 120 velocity vectors of the two-dimensional flow over a fixed time delay...with a wall cavity flameholder. In the HTV method, ArF excimer laser (193 nm) beams pass through a humid gas flow and dissociate H2O into H + OH to

Enzyme-linked electrochemical DNA ligation assay using magnetic beads.

PubMed

Stejskalová, Eva; Horáková, Petra; Vacek, Jan; Bowater, Richard P; Fojta, Miroslav

2014-07-01

DNA ligases are essential enzymes in all cells and have been proposed as targets for novel antibiotics. Efficient DNA ligase activity assays are thus required for applications in biomedical research. Here we present an enzyme-linked electrochemical assay based on two terminally tagged probes forming a nicked junction upon hybridization with a template DNA. Nicked DNA bearing a 5' biotin tag is immobilized on the surface of streptavidin-coated magnetic beads, and ligated product is detected via a 3' digoxigenin tag recognized by monoclonal antibody-alkaline phosphatase conjugate. Enzymatic conversion of napht-1-yl phosphate to napht-1-ol enables sensitive detection of the voltammetric signal on a pyrolytic graphite electrode. The technique was tested under optimal conditions and various situations limiting or precluding the ligation reaction (such as DNA substrates lacking 5'-phosphate or containing a base mismatch at the nick junction, or application of incompatible cofactor), and utilized for the analysis of the nick-joining activity of a range of recombinant Escherichia coli DNA ligase constructs. The novel technique provides a fast, versatile, specific, and sensitive electrochemical assay of DNA ligase activity.

Single-molecule imaging of cytoplasmic dynein in vivo.

PubMed

Ananthanarayanan, Vaishnavi; Tolić, Iva M

2015-01-01

While early fluorescence microscopy experiments employing fluorescent probes afforded snapshots of the cell, the power of live-cell microscopy is required to understand complex dynamics in biological processes. The first successful cloning of green fluorescent protein in the 1990s paved the way for development of approaches that we now utilize for visualization in a living cell. In this chapter, we discuss a technique to observe fluorescently tagged single molecules in fission yeast. With a few simple modifications to the established total internal reflection fluorescence microscopy, cytoplasmic dynein molecules in the cytoplasm and on the microtubules can be visualized and their intracellular dynamics can be studied. We illustrate a technique to study motor behavior, which is not apparent in conventional ensemble studies of motors. In general, this technique can be employed to study single-molecule dynamics of fluorescently tagged proteins in the cell interior. Copyright © 2015 Elsevier Inc. All rights reserved.

Integrated model-based retargeting and optical proximity correction

NASA Astrophysics Data System (ADS)

Agarwal, Kanak B.; Banerjee, Shayak

2011-04-01

Conventional resolution enhancement techniques (RET) are becoming increasingly inadequate at addressing the challenges of subwavelength lithography. In particular, features show high sensitivity to process variation in low-k1 lithography. Process variation aware RETs such as process-window OPC are becoming increasingly important to guarantee high lithographic yield, but such techniques suffer from high runtime impact. An alternative to PWOPC is to perform retargeting, which is a rule-assisted modification of target layout shapes to improve their process window. However, rule-based retargeting is not a scalable technique since rules cannot cover the entire search space of two-dimensional shape configurations, especially with technology scaling. In this paper, we propose to integrate the processes of retargeting and optical proximity correction (OPC). We utilize the normalized image log slope (NILS) metric, which is available at no extra computational cost during OPC. We use NILS to guide dynamic target modification between iterations of OPC. We utilize the NILS tagging capabilities of Calibre TCL scripting to identify fragments with low NILS. We then perform NILS binning to assign different magnitude of retargeting to different NILS bins. NILS is determined both for width, to identify regions of pinching, and space, to locate regions of potential bridging. We develop an integrated flow for 1x metal lines (M1) which exhibits lesser lithographic hotspots compared to a flow with just OPC and no retargeting. We also observe cases where hotspots that existed in the rule-based retargeting flow are fixed using our methodology. We finally also demonstrate that such a retargeting methodology does not significantly alter design properties by electrically simulating a latch layout before and after retargeting. We observe less than 1% impact on latch Clk-Q and D-Q delays post-retargeting, which makes this methodology an attractive one for use in improving shape process windows without perturbing designed values.

Distinguishing aspartic and isoaspartic acids in peptides by several mass spectrometric fragmentation methods

PubMed Central

DeGraan-Weber, Nick; Zhang, Jun; Reilly, James P.

2016-01-01

Six ion fragmentation techniques that can distinguish aspartic acid from its isomer, isoaspartic acid, were compared. MALDI post source decay (PSD), MALDI 157 nm photodissociation, TMPP charge tagging in PSD and photodissociation, ESI collision-induced dissociation (CID), electron transfer dissociation (ETD), and free-radical initiated peptide sequencing (FRIPS) with CID were applied to peptides containing either aspartic or isoaspartic acid. Diagnostic ions, such as the y-46 and b+H2O, are present in PSD, photodissociation, and charge tagging. c•+57 and z-57 ions are observed in ETD and FRIPS experiments. For some molecules, aspartic and isoaspartic acid yield ion fragments with significantly different intensities. ETD and charge tagging appear to be most effective at distinguishing these residues. PMID:27613306

Investigations of belukha whales in coastal waters of western and northern Alaska, 1982-1983: marking and tracking of whales in Bristol Bay

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

Frost, K.J.; Lowry, L.F.; Nelson, R.R.

1983-12-01

A 2-year study was conducted in Bristol Bay, Alaska, to develop and test techniques for marking belukha whales with visual and radio tags. Information was also gathered on belukha distribution and abundance, foods and feeding, and rates and causes of mortality. Two types of radio packages were developed: an OAR backpack designed to be bolted through the dorsal ridge, and a Telonics barnacle tag with an umbrella-stake attachment. Testing of tags and attachments revealed that the more-powerful OAR radio could be received at longer distances and lower antenna heights, and the the umbrella-stake attachment penetrated too deeply for reliable usemore » on belukhas.« less

Weighted SAW reflector gratings for orthogonal frequency coded SAW tags and sensors

NASA Technical Reports Server (NTRS)

Puccio, Derek (Inventor); Malocha, Donald (Inventor)

2011-01-01

Weighted surface acoustic wave reflector gratings for coding identification tags and sensors to enable unique sensor operation and identification for a multi-sensor environment. In an embodiment, the weighted reflectors are variable while in another embodiment the reflector gratings are apodized. The weighting technique allows the designer to decrease reflectively and allows for more chips to be implemented in a device and, consequently, more coding diversity. As a result, more tags and sensors can be implemented using a given bandwidth when compared with uniform reflectors. Use of weighted reflector gratings with OFC makes various phase shifting schemes possible, such as in-phase and quadrature implementations of coded waveforms resulting in reduced device size and increased coding.

Viral infection of the marine alga Emiliania huxleyi triggers lipidome remodeling and induces the production of highly saturated triacylglycerol.

PubMed

Malitsky, Sergey; Ziv, Carmit; Rosenwasser, Shilo; Zheng, Shuning; Schatz, Daniella; Porat, Ziv; Ben-Dor, Shifra; Aharoni, Asaph; Vardi, Assaf

2016-04-01

Viruses that infect marine photosynthetic microorganisms are major ecological and evolutionary drivers of microbial food webs, estimated to turn over more than a quarter of the total photosynthetically fixed carbon. Viral infection of the bloom-forming microalga Emiliania huxleyi induces the rapid remodeling of host primary metabolism, targeted towards fatty acid metabolism. We applied a liquid chromatography-mass spectrometry (LC-MS)-based lipidomics approach combined with imaging flow cytometry and gene expression profiling to explore the impact of viral-induced metabolic reprogramming on lipid composition. Lytic viral infection led to remodeling of the cellular lipidome, by predominantly inducing the biosynthesis of highly saturated triacylglycerols (TAGs), coupled with a significant accumulation of neutral lipids within lipid droplets. Furthermore, TAGs were found to be a major component (77%) of the lipidome of isolated virions. Interestingly, viral-induced TAGs were significantly more saturated than TAGs produced under nitrogen starvation. This study highlights TAGs as major products of the viral-induced metabolic reprogramming during the host-virus interaction and indicates a selective mode of membrane recruitment during viral assembly, possibly by budding of the virus from specialized subcellular compartments. These findings provide novel insights into the role of viruses infecting microalgae in regulating metabolism and energy transfer in the marine environment and suggest their possible biotechnological application in biofuel production. © 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.

Ultrasonically Encoded Photoacoustic Flowgraphy in Biological Tissue

NASA Astrophysics Data System (ADS)

Wang, Lidai; Xia, Jun; Yao, Junjie; Maslov, Konstantin I.; Wang, Lihong V.

2013-11-01

Blood flow speed is an important functional parameter. Doppler ultrasound flowmetry lacks sufficient sensitivity to slow blood flow (several to tens of millimeters per second) in deep tissue. To address this challenge, we developed ultrasonically encoded photoacoustic flowgraphy combining ultrasonic thermal tagging with photoacoustic imaging. Focused ultrasound generates a confined heat source in acoustically absorptive fluid. Thermal waves propagate with the flow and are directly visualized in pseudo color using photoacoustic computed tomography. The Doppler shift is employed to calculate the flow speed. This method requires only acoustic and optical absorption, and thus is applicable to continuous fluid. A blood flow speed as low as 0.24mm·s-1 was successfully measured. Deep blood flow imaging was experimentally demonstrated under 5-mm-thick chicken breast tissue.

Differently localized lysophosphatidic acid acyltransferases crucial for triacylglycerol biosynthesis in the oleaginous alga Nannochloropsis.

PubMed

Nobusawa, Takashi; Hori, Koichi; Mori, Hiroshi; Kurokawa, Ken; Ohta, Hiroyuki

2017-05-01

The production of renewable bioenergy will be necessary to meet rising global fossil fuel demands. Members of the marine microalgae genus Nannochloropsis produce large quantities of oils (triacylglycerols; TAGs), and this genus is regarded as one of the most promising for biodiesel production. Recent genome sequencing and transcriptomic studies on Nannochloropsis have provided a foundation for understanding its oleaginous trait, but the mechanism underlying oil accumulation remains to be clarified. Here we report Nannochloropsis knock-out strains of four extraplastidic lysophosphatidic acid acyltransferases (LPAT1-LPAT4) that catalyze a major de novo biosynthetic step of TAGs and membrane lipids. We found that the four LPATs are differently involved in lipid metabolic flow in Nannochloropsis. Double knock-outs among the LPATs revealed the pivotal LPATs for TAG biosynthesis, and localization analysis indicated that the stramenopile-specific LPATs (LPAT3 and LPAT4) associated with TAG synthesis reside at the perimeter of lipid droplets. No homologous region has been found with other lipid droplet-associated proteins, however. Lipid droplets are an organelle found in nearly all organisms, and recently they were shown to play important roles in cellular metabolism and signaling. Our results provide direct evidence for the importance of the perimeter of lipid droplet in TAG synthesis in addition to its known role in maintaining TAG stability, and these findings suggest that the oleaginous trait of Nannochloropsis is enabled by the acquisition of LPATs at the perimeter of lipid droplets. © 2017 The Authors. The Plant Journal published by John Wiley & Sons Ltd and Society for Experimental Biology.

Monitoring of adult Lost River and shortnose suckers in Clear Lake Reservoir, California, 2008–2010

USGS Publications Warehouse

Hewitt, David A.; Hayes, Brian S.

2013-01-01

Problems with inferring status and population dynamics from size composition data can be overcome by a robust capture-recapture program that follows the histories of PIT-tagged individuals. Inferences from such a program are currently hindered by poor detection rates during spawning seasons with low flows in Willow Creek, which indicate that a key assumption of capture-recapture models is violated. We suggest that the most straightforward solution to this issue would be to collect detection data during the spawning season using remote PIT tag antennas in the strait between the west and east lobes of the lake.

A method to analyze molecular tagging velocimetry data using the Hough transform.

PubMed

Sanchez-Gonzalez, R; McManamen, B; Bowersox, R D W; North, S W

2015-10-01

The development of a method to analyze molecular tagging velocimetry data based on the Hough transform is presented. This method, based on line fitting, parameterizes the grid lines "written" into a flowfield. Initial proof-of-principle illustration of this method was performed to obtain two-component velocity measurements in the wake of a cylinder in a Mach 4.6 flow, using a data set derived from computational fluid dynamics simulations. The Hough transform is attractive for molecular tagging velocimetry applications since it is capable of discriminating spurious features that can have a biasing effect in the fitting process. Assessment of the precision and accuracy of the method were also performed to show the dependence on analysis window size and signal-to-noise levels. The accuracy of this Hough transform-based method to quantify intersection displacements was determined to be comparable to cross-correlation methods. The employed line parameterization avoids the assumption of linearity in the vicinity of each intersection, which is important in the limit of drastic grid deformations resulting from large velocity gradients common in high-speed flow applications. This Hough transform method has the potential to enable the direct and spatially accurate measurement of local vorticity, which is important in applications involving turbulent flowfields. Finally, two-component velocity determinations using the Hough transform from experimentally obtained images are presented, demonstrating the feasibility of the proposed analysis method.

Measuring the performance of two stationary interrogation systems for detecting downstream and upstream movement of PIT-tagged salmonids

USGS Publications Warehouse

Connolly, P.J.; Jezorek, I.G.; Martens, K.D.; Prentice, E.F.

2008-01-01

We tested the performance of two stationary interrogation systems designed for detecting the movement of fish with passive integrated transponder (PIT) tags. These systems allowed us to determine the direction of fish movement with high detection efficiency and high precision in a dynamic stream environment. We describe an indirect method for deriving an estimate for detection efficiency and the associated variance that does not rely on a known number of fish passing the system. By using six antennas arranged in a longitudinal series of three arrays, we attained detection efficiencies for downstream- and upstream-moving fish exceeding 96% during high-flow periods and approached 100% during low-flow periods for the two interrogation systems we tested. Because these systems did not rely on structural components, such as bridges or culverts, they were readily adaptable to remote, natural stream sites. Because of built-in redundancy, these systems were able to perform even with a loss of one or more antennas owing to dislodgement or electrical failure. However, the reduction in redundancy resulted in decreased efficiency and precision and the potential loss of ability to determine the direction of fish movement. What we learned about these systems should be applicable to a wide variety of other antenna configurations and to other types of PIT tags and transceivers.

Partial entrainment of gravel bars during floods

USGS Publications Warehouse

Konrad, Christopher P.; Booth, Derek B.; Burges, Stephen J.; Montgomery, David R.

2002-01-01

Spatial patterns of bed material entrainment by floods were documented at seven gravel bars using arrays of metal washers (bed tags) placed in the streambed. The observed patterns were used to test a general stochastic model that bed material entrainment is a spatially independent, random process where the probability of entrainment is uniform over a gravel bar and a function of the peak dimensionless shear stress τ0* of the flood. The fraction of tags missing from a gravel bar during a flood, or partial entrainment, had an approximately normal distribution with respect to τ0* with a mean value (50% of the tags entrained) of 0.085 and standard deviation of 0.022 (root‐mean‐square error of 0.09). Variation in partial entrainment for a given τ0* demonstrated the effects of flow conditioning on bed strength, with lower values of partial entrainment after intermediate magnitude floods (0.065 < τ0*< 0.08) than after higher magnitude floods. Although the probability of bed material entrainment was approximately uniform over a gravel bar during individual floods and independent from flood to flood, regions of preferential stability and instability emerged at some bars over the course of a wet season. Deviations from spatially uniform and independent bed material entrainment were most pronounced for reaches with varied flow and in consecutive floods with small to intermediate magnitudes.

Estimating survival of radio-tagged birds

USGS Publications Warehouse

Bunck, C.M.; Pollock, K.H.; Lebreton, J.-D.; North, P.M.

1993-01-01

Parametric and nonparametric methods for estimating survival of radio-tagged birds are described. The general assumptions of these methods are reviewed. An estimate based on the assumption of constant survival throughout the period is emphasized in the overview of parametric methods. Two nonparametric methods, the Kaplan-Meier estimate of the survival funcrion and the log rank test, are explained in detail The link between these nonparametric methods and traditional capture-recapture models is discussed aloag with considerations in designing studies that use telemetry techniques to estimate survival.

Iodine Tagging Velocimetry and Mechanism in the Hypersonic Near Wake of a MultiPurpose Crew Vehicle

NASA Technical Reports Server (NTRS)

Balla, R. Jeffrey

2013-01-01

This study demonstrates a new molecular tagging velocimetry (MTV) method for velocity measurements of high speed flow. It demonstrates offbody Iodine Tagging Velocimetry (ITV) in the hypersonic near wake of a MultiPurpose Crew Vehicle (MPCV) model. Experiments are performed in the NASA-Langley 31-inch Mach 10 air wind tunnel. A 0.5% I2 / N2 mixture is seeded on the leeward backshell of the model using a pressure tap. I2 laser-induced fluorescence is excited along a 5.5 mm line using an ArF excimer laser near 193 nm. Results indicate I2 absorbs at least 2 photons to produce iodine ions and electrons. These recombine as the tagged region is displaced downstream to produce I (2P3/2) whose emission is monitored at 206 nm. Results at P0 = 2.41 MPa (350 psi), T0 = 990K, and 10 micro-sec transit times produce velocities from 630-820 m/sec across the I2 seeded jet at a distance of 38.2 mm (25.5 jet diameters) downstream from the jet orifice. Maximum wake jet velocities near the shear layer are 59% of freestream velocity.

A tag-based approach for high-throughput analysis of CCWGG methylation.

PubMed

Denisova, Oksana V; Chernov, Andrei V; Koledachkina, Tatyana Y; Matvienko, Nicholas I

2007-10-15

Non-CpG methylation occurring in the context of CNG sequences is found in plants at a large number of genomic loci. However, there is still little information available about non-CpG methylation in mammals. Efficient methods that would allow detection of scarcely localized methylated sites in small quantities of DNA are required to elucidate the biological role of non-CpG methylation in both plants and animals. In this study, we tested a new whole genome approach to identify sites of CCWGG methylation (W is A or T), a particular case of CNG methylation, in genomic DNA. This technique is based on digestion of DNAs with methylation-sensitive restriction endonucleases EcoRII-C and AjnI. Short DNAs flanking methylated CCWGG sites (tags) are selectively purified and assembled in tandem arrays of up to nine tags. This allows high-throughput sequencing of tags, identification of flanking regions, and their exact positions in the genome. In this study, we tested specificity and efficiency of the approach.

SPlinted Ligation Adapter Tagging (SPLAT), a novel library preparation method for whole genome bisulphite sequencing

PubMed Central

Manlig, Erika; Wahlberg, Per

2017-01-01

Abstract Sodium bisulphite treatment of DNA combined with next generation sequencing (NGS) is a powerful combination for the interrogation of genome-wide DNA methylation profiles. Library preparation for whole genome bisulphite sequencing (WGBS) is challenging due to side effects of the bisulphite treatment, which leads to extensive DNA damage. Recently, a new generation of methods for bisulphite sequencing library preparation have been devised. They are based on initial bisulphite treatment of the DNA, followed by adaptor tagging of single stranded DNA fragments, and enable WGBS using low quantities of input DNA. In this study, we present a novel approach for quick and cost effective WGBS library preparation that is based on splinted adaptor tagging (SPLAT) of bisulphite-converted single-stranded DNA. Moreover, we validate SPLAT against three commercially available WGBS library preparation techniques, two of which are based on bisulphite treatment prior to adaptor tagging and one is a conventional WGBS method. PMID:27899585

Short range spread-spectrum radiolocation system and method

DOEpatents

Smith, Stephen F.

2003-04-29

A short range radiolocation system and associated methods that allow the location of an item, such as equipment, containers, pallets, vehicles, or personnel, within a defined area. A small, battery powered, self-contained tag is provided to an item to be located. The tag includes a spread-spectrum transmitter that transmits a spread-spectrum code and identification information. A plurality of receivers positioned about the area receive signals from a transmitting tag. The position of the tag, and hence the item, is located by triangulation. The system employs three different ranging techniques for providing coarse, intermediate, and fine spatial position resolution. Coarse positioning information is provided by use of direct-sequence code phase transmitted as a spread-spectrum signal. Intermediate positioning information is provided by the use of a difference signal transmitted with the direct-sequence spread-spectrum code. Fine positioning information is provided by use of carrier phase measurements. An algorithm is employed to combine the three data sets to provide accurate location measurements.

Multi-processor including data flow accelerator module

DOEpatents

Davidson, George S.; Pierce, Paul E.

1990-01-01

An accelerator module for a data flow computer includes an intelligent memory. The module is added to a multiprocessor arrangement and uses a shared tagged memory architecture in the data flow computer. The intelligent memory module assigns locations for holding data values in correspondence with arcs leading to a node in a data dependency graph. Each primitive computation is associated with a corresponding memory cell, including a number of slots for operands needed to execute a primitive computation, a primitive identifying pointer, and linking slots for distributing the result of the cell computation to other cells requiring that result as an operand. Circuitry is provided for utilizing tag bits to determine automatically when all operands required by a processor are available and for scheduling the primitive for execution in a queue. Each memory cell of the module may be associated with any of the primitives, and the particular primitive to be executed by the processor associated with the cell is identified by providing an index, such as the cell number for the primitive, to the primitive lookup table of starting addresses. The module thus serves to perform functions previously performed by a number of sections of data flow architectures and coexists with conventional shared memory therein. A multiprocessing system including the module operates in a hybrid mode, wherein the same processing modules are used to perform some processing in a sequential mode, under immediate control of an operating system, while performing other processing in a data flow mode.

An integrated PCR colony hybridization approach to screen cDNA libraries for full-length coding sequences.

PubMed

Pollier, Jacob; González-Guzmán, Miguel; Ardiles-Diaz, Wilson; Geelen, Danny; Goossens, Alain

2011-01-01

cDNA-Amplified Fragment Length Polymorphism (cDNA-AFLP) is a commonly used technique for genome-wide expression analysis that does not require prior sequence knowledge. Typically, quantitative expression data and sequence information are obtained for a large number of differentially expressed gene tags. However, most of the gene tags do not correspond to full-length (FL) coding sequences, which is a prerequisite for subsequent functional analysis. A medium-throughput screening strategy, based on integration of polymerase chain reaction (PCR) and colony hybridization, was developed that allows in parallel screening of a cDNA library for FL clones corresponding to incomplete cDNAs. The method was applied to screen for the FL open reading frames of a selection of 163 cDNA-AFLP tags from three different medicinal plants, leading to the identification of 109 (67%) FL clones. Furthermore, the protocol allows for the use of multiple probes in a single hybridization event, thus significantly increasing the throughput when screening for rare transcripts. The presented strategy offers an efficient method for the conversion of incomplete expressed sequence tags (ESTs), such as cDNA-AFLP tags, to FL-coding sequences.

Augmenting matrix factorization technique with the combination of tags and genres

NASA Astrophysics Data System (ADS)

Ma, Tinghuai; Suo, Xiafei; Zhou, Jinjuan; Tang, Meili; Guan, Donghai; Tian, Yuan; Al-Dhelaan, Abdullah; Al-Rodhaan, Mznah

2016-11-01

Recommender systems play an important role in our daily life and are becoming popular tools for users to find what they are really interested in. Matrix factorization methods, which are popular recommendation methods, have gained high attention these years. With the rapid growth of the Internet, lots of information has been created, like social network information, tags and so on. Along with these, a few matrix factorization approaches have been proposed which incorporate the personalized information of users or items. However, except for ratings, most of the matrix factorization models have utilized only one kind of information to understand users' interests. Considering the sparsity of information, in this paper, we try to investigate the combination of different information, like tags and genres, to reveal users' interests accurately. With regard to the generalization of genres, a constraint is added when genres are utilized to find users' similar ;soulmates;. In addition, item regularizer is also considered based on latent semantic indexing (LSI) method with the item tags. Our experiments are conducted on two real datasets: Movielens dataset and Douban dataset. The experimental results demonstrate that the combination of tags and genres is really helpful to reveal users' interests.

A SNAP-Tagged Derivative of HIV-1—A Versatile Tool to Study Virus-Cell Interactions

PubMed Central

Eckhardt, Manon; Anders, Maria; Muranyi, Walter; Heilemann, Mike; Krijnse-Locker, Jacomine; Müller, Barbara

2011-01-01

Fluorescently labeled human immunodeficiency virus (HIV) derivatives, combined with the use of advanced fluorescence microscopy techniques, allow the direct visualization of dynamic events and individual steps in the viral life cycle. HIV proteins tagged with fluorescent proteins (FPs) have been successfully used for live-cell imaging analyses of HIV-cell interactions. However, FPs display limitations with respect to their physicochemical properties, and their maturation kinetics. Furthermore, several independent FP-tagged constructs have to be cloned and characterized in order to obtain spectral variations suitable for multi-color imaging setups. In contrast, the so-called SNAP-tag represents a genetically encoded non-fluorescent tag which mediates specific covalent coupling to fluorescent substrate molecules in a self-labeling reaction. Fusion of the SNAP-tag to the protein of interest allows specific labeling of the fusion protein with a variety of synthetic dyes, thereby offering enhanced flexibility for fluorescence imaging approaches. Here we describe the construction and characterization of the HIV derivative HIVSNAP, which carries the SNAP-tag as an additional domain within the viral structural polyprotein Gag. Introduction of the tag close to the C-terminus of the matrix domain of Gag did not interfere with particle assembly, release or proteolytic virus maturation. The modified virions were infectious and could be propagated in tissue culture, albeit with reduced replication capacity. Insertion of the SNAP domain within Gag allowed specific staining of the viral polyprotein in the context of virus producing cells using a SNAP reactive dye as well as the visualization of individual virions and viral budding sites by stochastic optical reconstruction microscopy. Thus, HIVSNAP represents a versatile tool which expands the possibilities for the analysis of HIV-cell interactions using live cell imaging and sub-diffraction fluorescence microscopy. PMID:21799764

Use of portable antennas to estimate abundance of PIT-tagged fish in small streams: Factors affecting detection probability

USGS Publications Warehouse

O'Donnell, Matthew J.; Horton, Gregg E.; Letcher, Benjamin H.

2010-01-01

Portable passive integrated transponder (PIT) tag antenna systems can be valuable in providing reliable estimates of the abundance of tagged Atlantic salmon Salmo salar in small streams under a wide range of conditions. We developed and employed PIT tag antenna wand techniques in two controlled experiments and an additional case study to examine the factors that influenced our ability to estimate population size. We used Pollock's robust-design capture–mark–recapture model to obtain estimates of the probability of first detection (p), the probability of redetection (c), and abundance (N) in the two controlled experiments. First, we conducted an experiment in which tags were hidden in fixed locations. Although p and c varied among the three observers and among the three passes that each observer conducted, the estimates of N were identical to the true values and did not vary among observers. In the second experiment using free-swimming tagged fish, p and c varied among passes and time of day. Additionally, estimates of N varied between day and night and among age-classes but were within 10% of the true population size. In the case study, we used the Cormack–Jolly–Seber model to examine the variation in p, and we compared counts of tagged fish found with the antenna wand with counts collected via electrofishing. In that study, we found that although p varied for age-classes, sample dates, and time of day, antenna and electrofishing estimates of N were similar, indicating that population size can be reliably estimated via PIT tag antenna wands. However, factors such as the observer, time of day, age of fish, and stream discharge can influence the initial and subsequent detection probabilities.

Dual Parallel Mass Spectrometry (LC1/MS2 and LC2/MS2) for Lipid and Vitamin D Analysis

USDA-ARS?s Scientific Manuscript database

Atmospheric pressure chemical ionization (APCI) mass spectrometry (MS) and electrospray ionization (ESI) MS are complementary techniques that provide different types of information for lipids such as triacylglycerols (TAGs), phospholipids, and fat-soluble vitamins. Since no one technique is by itsel...

Three Component Velocity and Acceleration Measurement Using FLEET

NASA Technical Reports Server (NTRS)

Danehy, Paul M.; Bathel, Brett F.; Calvert, Nathan; Dogariu, Arthur; Miles, Richard P.

2014-01-01

The femtosecond laser electronic excitation and tagging (FLEET) method has been used to measure three components of velocity and acceleration for the first time. A jet of pure N2 issuing into atmospheric pressure air was probed by the FLEET system. The femtosecond laser was focused down to a point to create a small measurement volume in the flow. The long-lived lifetime of this fluorescence was used to measure the location of the tagged particles at different times. Simultaneous images of the flow were taken from two orthogonal views using a mirror assembly and a single intensified CCD camera, allowing two components of velocity to be measured in each view. These different velocity components were combined to determine three orthogonal velocity components. The differences between subsequent velocity components could be used to measure the acceleration. Velocity accuracy and precision were roughly estimated to be +/-4 m/s and +/-10 m/s respectively. These errors were small compared to the approx. 100 m/s velocity of the subsonic jet studied.

Specific solubilization of impurities in culture media: Arg solution improves purification of pH-responsive tag CspB50 with Teriparatide.

PubMed

Oki, Shogo; Nonaka, Takahiro; Shiraki, Kentaro

2018-06-01

Protein purification using non-chromatographic methods is a simple technique that avoids costly resin. Recently, a cell surface protein B (CspB) tag has been developed for a pH-responsive tag for protein purification by solid-liquid separation. Proteins fused with the CspB tag show reversible insolubilization at acidic pH that can be used in solid-liquid separation for protein purification. However, brown-color impurities from co-precipitation hamper further analysis of the target proteins. In this study, we investigated the effect of additives on the co-precipitation of CspB-tagged Teriparatide (CspB50TEV-Teriparatide) expressed in Corynebacterium glutamicum and associated impurities. Arginine (Arg) at 1.0 M was found to be the most effective additive for removing impurities, particularly carotenoids and nucleic acids. Furthermore, all impurities detected in the fluorescence and absorbance spectra were successfully removed by the repetition of precipitation-redissolution in the Arg solution. The precipitation yield of the CspB50TEV-Teriparatide did not change with the addition of Arg and the repetition of the precipitation-redissolution process. Collectively, our findings indicate that the specific desorption of π-electron rich compounds by Arg may be useful in conjunction with the pH-responsive CspB tag for solid-liquid protein purification. Copyright © 2018 Elsevier Inc. All rights reserved.

Utility of biological sensor tags in animal conservation.

PubMed

Wilson, A D M; Wikelski, M; Wilson, R P; Cooke, S J

2015-08-01

Electronic tags (both biotelemetry and biologging platforms) have informed conservation and resource management policy and practice by providing vital information on the spatial ecology of animals and their environments. However, the extent of the contribution of biological sensors (within electronic tags) that measure an animal's state (e.g., heart rate, body temperature, and details of locomotion and energetics) is less clear. A literature review revealed that, despite a growing number of commercially available state sensor tags and enormous application potential for such devices in animal biology, there are relatively few examples of their application to conservation. Existing applications fell under 4 main themes: quantifying disturbance (e.g., ecotourism, vehicular and aircraft traffic), examining the effects of environmental change (e.g., climate change), understanding the consequences of habitat use and selection, and estimating energy expenditure. We also identified several other ways in which sensor tags could benefit conservation, such as determining the potential efficacy of management interventions. With increasing sensor diversity of commercially available platforms, less invasive attachment techniques, smaller device sizes, and more researchers embracing such technology, we suggest that biological sensor tags be considered a part of the necessary toolbox for conservation. This approach can measure (in real time) the state of free-ranging animals and thus provide managers with objective, timely, relevant, and accurate data to inform policy and decision making. © 2015 Society for Conservation Biology.

Survival and migration behavior of juvenile salmonids at McNary Dam, 2004, Final report of research

USGS Publications Warehouse

Perry, Russell W.; Braatz, Amy C.; Fielding, Scott D.; Lucchesi, Joel N.; Plumb, John M.; Adams, Noah S.; Rondorf, Dennis W.

2005-01-01

During 2004, the USGS Columbia River Research Laboratory conducted a study at McNary Dam using radio telemetry to estimate passage and survival parameters of juvenile salmonids. Our primary objective was to estimate these parameters under ambient environmental and operational conditions, and thus project-wide treatments were not implemented. The primary dam operation consisted of “biop” spill, where spill occurred at night between 1800 and 0600 hours, and no spill occurred between 0600 and 1800 hours for the majority of our study period. During the spring study period, we radio-tagged and released 1,896 yearling Chinook salmon and 1,888 juvenile steelhead. During the summer study period, we radio-tagged and released 1,919 subyearling Chinook salmon. All fish were tagged using gastric techniques to implant transmitters weighing 1.58 g for yearling Chinook salmon, 1.93 g for juvenile steelhead, and 0.96 g for subyearling Chinook salmon. Minimum fish sizes were based on a 6.5% tag:fish weight ratio, and the size of tagged fish represented about 91%, 100%, and 17% of the population, respectively for yearling Chinook salmon, juvenile steelhead, and subyearling Chinook salmon. About 60% of radio-tagged fish were released 10 km upstream of McNary Dam at Hat Rock State Park, Oregon, with the remainder released as control groups 400 m downstream of the dam.

Development of novel wireless sensor for food quality detection

NASA Astrophysics Data System (ADS)

Son Nguyen, Dat; Ngan Le, Nguyen; Phat Lam, Tan; Fribourg-Blanc, Eric; Chien Dang, Mau; Tedjini, Smail

2015-12-01

In this paper we present a wireless sensor for the monitoring of food quality. We integrate sensing capability into ultrahigh frequency (UHF) radio-frequency identification (RFID) tags through the relationship between the physical read-range and permittivity of the object we label with the RFID tags. Using the known variations of food permittivity as a function of time, we can detect the contamination time at which a food product becomes unacceptable for consumption based on the measurement of read-range with the as-designed sensing tags. This low-cost UHF RFID passive sensor was designed and experimentally tested on beef, pork, and cheese with the same storage conditions as in supermarkets. The agreement between the experimental and simulation results show the potential of this technique for practical application in food-quality tracking.

Proposed measurement of tagged deep inelastic scattering in Hall A of Jefferson lab

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

Montgomery, Rachel; Annand, John; Dutta, Dipangkar

2017-03-01

A tagged deep inelastic scattering (TDIS) experiment is planned for Hall A of Jefferson Lab, which will probe the mesonic content of the nucleon directly. Low momentum recoiling (and spectator) protons will be measured in coincidence with electrons scattered in a deep inelastic regime from hydrogen (and deuterium) targets, covering kinematics of 8 < W2 < 18 GeV2, 1 < Q2 < 3 (GeV/c)2 and 0:05 < x < 0:2. The tagging technique will help identify scattering from partons in the meson cloud and provide access to the pion structure function via the Sullivan process. The experiment will yield themore » first TDIS results in the valence regime, for both proton and neutron targets. We present here an overview of the experiment.« less

General Framework for Animal Food Safety Traceability Using GS1 and RFID

NASA Astrophysics Data System (ADS)

Cao, Weizhu; Zheng, Limin; Zhu, Hong; Wu, Ping

GS1 is global traceability standard, which is composed by the encoding system (EAN/UCC, EPC), the data carriers identified automatically (bar codes, RFID), electronic data interchange standards (EDI, XML). RFID is a non-contact, multi-objective automatic identification technique. Tracing of source food, standardization of RFID tags, sharing of dynamic data are problems to solve urgently for recent traceability systems. The paper designed general framework for animal food safety traceability using GS1 and RFID. This framework uses RFID tags encoding with EPCglobal tag data standards. Each information server has access tier, business tier and resource tier. These servers are heterogeneous and distributed, providing user access interfaces by SOAP or HTTP protocols. For sharing dynamic data, discovery service and object name service are used to locate dynamic distributed information servers.

Leveraging algal omics to reveal potential targets for augmenting TAG accumulation

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

Arora, Neha; Pienkos, Philip T.; Pruthi, Vikas

Ongoing global efforts to commercialize microalgal biofuels have expedited the use of multi-omics techniques to gain insights into lipid biosynthetic pathways. Functional genomics analyses have recently been employed to complement existing sequence-level omics studies, shedding light on the dynamics of lipid synthesis and its interplay with other cellular metabolic pathways, thus revealing possible targets for metabolic engineering. Here, we review the current status of algal omics studies to reveal potential targets to augment TAG accumulation in various microalgae. Here, this review specifically aims to examine and catalog systems level data related to stress-induced TAG accumulation in oleaginous microalgae and informmore » future metabolic engineering strategies to develop strains with enhanced bioproductivity, which could pave a path for sustainable green energy.« less

Leveraging Algal Omics to Reveal Potential Targets for Augmenting TAG Accumulation

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

Guarnieri, Michael T; Pienkos, Philip T; Arora, Neha

2018-04-18

Ongoing global efforts to commercialize microalgal biofuels have expedited the use of multi-omics techniques to gain insights into lipid biosynthetic pathways. Functional genomics analyses have recently been employed to complement existing sequence-level omics studies, shedding light on the dynamics of lipid synthesis and its interplay with other cellular metabolic pathways, thus revealing possible targets for metabolic engineering. Here, we review the current status of algal omics studies to reveal potential targets to augment TAG accumulation in various microalgae. This review specifically aims to examine and catalog systems level data related to stress-induced TAG accumulation in oleaginous microalgae and inform futuremore » metabolic engineering strategies to develop strains with enhanced bioproductivity, which could pave a path for sustainable green energy.« less

Distinguishing Aspartic and Isoaspartic Acids in Peptides by Several Mass Spectrometric Fragmentation Methods

NASA Astrophysics Data System (ADS)

DeGraan-Weber, Nick; Zhang, Jun; Reilly, James P.

2016-12-01

Six ion fragmentation techniques that can distinguish aspartic acid from its isomer, isoaspartic acid, were compared. MALDI post-source decay (PSD), MALDI 157 nm photodissociation, tris(2,4,6-trimethoxyphenyl)phosphonium bromide (TMPP) charge tagging in PSD and photodissociation, ESI collision-induced dissociation (CID), electron transfer dissociation (ETD), and free-radical initiated peptide sequencing (FRIPS) with CID were applied to peptides containing either aspartic or isoaspartic acid. Diagnostic ions, such as the y-46 and b+H2O, are present in PSD, photodissociation, and charge tagging. c•+57 and z-57 ions are observed in ETD and FRIPS experiments. For some molecules, aspartic and isoaspartic acid yield ion fragments with significantly different intensities. ETD and charge tagging appear to be most effective at distinguishing these residues.

Structural identification of triacylglycerol isomers using electron impact excitation of ions from organics (EIEIO).

PubMed

Baba, Takashi; Campbell, J Larry; Le Blanc, J C Yves; Baker, Paul R S

2016-11-01

Electron-induced dissociation or electron impact excitation of ions from organics (EIEIO) was applied to triacylglycerols (TAGs) for in-depth molecular structure analysis using MS. In EIEIO, energetic electrons (∼10 eV) fragmented TAG ions to allow for regioisomeric assignment of identified acyl groups at the sn-2 or sn-1/3 positions of the glycerol backbone. In addition, carbon-carbon double bond locations within the acyl chains could also be assigned by EIEIO. Beyond the analysis of lipid standards, this technique was applied to edible oils and natural lipid extracts to demonstrate the power of this method to provide in-depth structural elucidation of TAG molecular species. Copyright © 2016 by the American Society for Biochemistry and Molecular Biology, Inc.

Leveraging algal omics to reveal potential targets for augmenting TAG accumulation

DOE PAGES

Arora, Neha; Pienkos, Philip T.; Pruthi, Vikas; ...

2018-04-18

Ongoing global efforts to commercialize microalgal biofuels have expedited the use of multi-omics techniques to gain insights into lipid biosynthetic pathways. Functional genomics analyses have recently been employed to complement existing sequence-level omics studies, shedding light on the dynamics of lipid synthesis and its interplay with other cellular metabolic pathways, thus revealing possible targets for metabolic engineering. Here, we review the current status of algal omics studies to reveal potential targets to augment TAG accumulation in various microalgae. Here, this review specifically aims to examine and catalog systems level data related to stress-induced TAG accumulation in oleaginous microalgae and informmore » future metabolic engineering strategies to develop strains with enhanced bioproductivity, which could pave a path for sustainable green energy.« less

Leveraging algal omics to reveal potential targets for augmenting TAG accumulation.

PubMed

Arora, Neha; Pienkos, Philip T; Pruthi, Vikas; Poluri, Krishna Mohan; Guarnieri, Michael T

2018-04-18

Ongoing global efforts to commercialize microalgal biofuels have expedited the use of multi-omics techniques to gain insights into lipid biosynthetic pathways. Functional genomics analyses have recently been employed to complement existing sequence-level omics studies, shedding light on the dynamics of lipid synthesis and its interplay with other cellular metabolic pathways, thus revealing possible targets for metabolic engineering. Here, we review the current status of algal omics studies to reveal potential targets to augment TAG accumulation in various microalgae. This review specifically aims to examine and catalog systems level data related to stress-induced TAG accumulation in oleaginous microalgae and inform future metabolic engineering strategies to develop strains with enhanced bioproductivity, which could pave a path for sustainable green energy. Copyright © 2018. Published by Elsevier Inc.

Modelling passive diastolic mechanics with quantitative MRI of cardiac structure and function.

PubMed

Wang, Vicky Y; Lam, H I; Ennis, Daniel B; Cowan, Brett R; Young, Alistair A; Nash, Martyn P

2009-10-01

The majority of patients with clinically diagnosed heart failure have normal systolic pump function and are commonly categorized as suffering from diastolic heart failure. The left ventricle (LV) remodels its structure and function to adapt to pathophysiological changes in geometry and loading conditions, which in turn can alter the passive ventricular mechanics. In order to better understand passive ventricular mechanics, a LV finite element (FE) model was customized to geometric data segmented from in vivo tagged magnetic resonance images (MRI) data and myofibre orientation derived from ex vivo diffusion tensor MRI (DTMRI) of a canine heart using nonlinear finite element fitting techniques. MRI tissue tagging enables quantitative evaluation of cardiac mechanical function with high spatial and temporal resolution, whilst the direction of maximum water diffusion in each voxel of a DTMRI directly corresponds to the local myocardial fibre orientation. Due to differences in myocardial geometry between in vivo and ex vivo imaging, myofibre orientations were mapped into the geometric FE model using host mesh fitting (a free form deformation technique). Pressure recordings, temporally synchronized to the tagging data, were used as the loading constraints to simulate the LV deformation during diastole. Simulation of diastolic LV mechanics allowed us to estimate the stiffness of the passive LV myocardium based on kinematic data obtained from tagged MRI. Integrated physiological modelling of this kind will allow more insight into mechanics of the LV on an individualized basis, thereby improving our understanding of the underlying structural basis of mechanical dysfunction under pathological conditions.

Seasonal and circadian biases in bird tracking with solar GPS-tags.

PubMed

Silva, Rafa; Afán, Isabel; Gil, Juan A; Bustamante, Javier

2017-01-01

Global Positioning System (GPS) tags are nowadays widely used in wildlife tracking. This geolocation technique can suffer from fix loss biases due to poor satellite GPS geometry, that result in tracking data gaps leading to wrong research conclusions. In addition, new solar-powered GPS tags deployed on birds can suffer from a new "battery drain bias" currently ignored in movement ecology analyses. We use a GPS tracking dataset of bearded vultures (Gypaetus barbatus), tracked for several years with solar GPS tags, to evaluate the causes and triggers of fix and data retrieval loss biases. We compare two models of solar GPS tags using different data retrieval systems (Argos vs GSM-GPRS), and programmed with different duty cycles. Neither of the models was able to accomplish the duty cycle programed initially. Fix and data retrieval loss rates were always greater than expected, and showed non-random gaps in GPS locations. Number of fixes per month of tracking was a bad criterion to identify tags with smaller biases. Fix-loss rates were four times higher due to battery drain than due to poor GPS satellite geometry. Both tag models were biased due to the uneven solar energy available for the recharge of the tag throughout the annual cycle, resulting in greater fix-loss rates in winter compared to summer. In addition, we suggest that the bias found along the diurnal cycle is linked to a complex three-factor interaction of bird flight behavior, topography and fix interval. More fixes were lost when vultures were perching compared to flying, in rugged versus flat topography. But long fix-intervals caused greater loss of fixes in dynamic (flying) versus static situations (perching). To conclude, we emphasize the importance of evaluating fix-loss bias in current tracking projects, and deploying GPS tags that allow remote duty cycle updates so that the most appropriate fix and data retrieval intervals can be selected.

Seasonal and circadian biases in bird tracking with solar GPS-tags

PubMed Central

Afán, Isabel; Gil, Juan A.; Bustamante, Javier

2017-01-01

Global Positioning System (GPS) tags are nowadays widely used in wildlife tracking. This geolocation technique can suffer from fix loss biases due to poor satellite GPS geometry, that result in tracking data gaps leading to wrong research conclusions. In addition, new solar-powered GPS tags deployed on birds can suffer from a new “battery drain bias” currently ignored in movement ecology analyses. We use a GPS tracking dataset of bearded vultures (Gypaetus barbatus), tracked for several years with solar GPS tags, to evaluate the causes and triggers of fix and data retrieval loss biases. We compare two models of solar GPS tags using different data retrieval systems (Argos vs GSM-GPRS), and programmed with different duty cycles. Neither of the models was able to accomplish the duty cycle programed initially. Fix and data retrieval loss rates were always greater than expected, and showed non-random gaps in GPS locations. Number of fixes per month of tracking was a bad criterion to identify tags with smaller biases. Fix-loss rates were four times higher due to battery drain than due to poor GPS satellite geometry. Both tag models were biased due to the uneven solar energy available for the recharge of the tag throughout the annual cycle, resulting in greater fix-loss rates in winter compared to summer. In addition, we suggest that the bias found along the diurnal cycle is linked to a complex three-factor interaction of bird flight behavior, topography and fix interval. More fixes were lost when vultures were perching compared to flying, in rugged versus flat topography. But long fix-intervals caused greater loss of fixes in dynamic (flying) versus static situations (perching). To conclude, we emphasize the importance of evaluating fix-loss bias in current tracking projects, and deploying GPS tags that allow remote duty cycle updates so that the most appropriate fix and data retrieval intervals can be selected. PMID:29020062

Radio-tracking manatees from land and space: tag design, implementation, and lessons learned from long-term study

USGS Publications Warehouse

Deutsch, C.J.; Bonde, R.K.; Reid, J.P.

1998-01-01

West Indian manatees (Trichechus manatus) were tracked along the Atlantic coast of Florida and Georgia (N = 83 manatees, n = 439 tag deployments, 1986-1996) and in eastern Puerto Rico (N = 8, n = 43, 1992-1996) using conventional and satellite-based radio-telemetry systems. A floating radio-tag, attached by a flexible tether to a padded belt around the base of the tail, enabled us to track manatees in saltwater environments. The tag incorporated VHF (very high frequency) and ultrasonic transmitters for field tracking and tag recovery, and an Argos satellite-monitored transmitter for remote tracking. We located each animal in the field about twice per week, received more than 60 000 good-quality Argos locations, and recovered tags in over 90% of deployments. The tag was designed to detach from the belt when entangled to prevent injury or drowning, and this often led to premature termination of tracking bouts. We had considerable success, however, in retagging belted manatees without recapture (97% of 392 retagging events). Most individuals were radio-tagged more than once (median = 3.0, maximum = 43) for a median total duration of 7.5 months (maximum = 6.8 yr). Data obtained through Argos have been valuable in addressing questions relating to long-distance movements, site fidelity, and identification of high-use areas. Fine-scale analyses of manatee habitat use and movements may require restricting the data set to the highest location quality or developing new analytical techniques to incorporate locational error. Field tracking provided useful ancillary data on life-history parameters, but sample sizes were small and survival estimates imprecise. Modification of the existing tag design to include Global Positioning System (GPS) functionality, with its finer spatial and temporal resolution, will offer new opportunities to address critical research and management problems facing this endangered species.

RFID tags as a direct tracer for water and sediment dynamics

NASA Astrophysics Data System (ADS)

Sommerer, Erik; Plate, Simon; Güntner, Andreas

2014-05-01

RFID (Radio Frequency IDentification) is a wireless automatic identification system to track objects with widespread application in industrial operations, but also selected applications in ecological research (animal tracking) and for hydro-sedimentological studies (sediment transport with RFID tags embedded in bedload material). In this study, for the first time, we test and apply RFID tags as a direct tracer to track water pathways, erosion patterns and sediment transport on the surface at the hillslope and headwater scale. The RFID system used here consists of tags with a size of 12 x 2 mm and a combination of mobile and stationary antennas. The transport pathways and velocities of the RFID tags can be individually assessed due to their unique identification numbers. The study area is a badland of easily erodible marls and carbonates located in the Villacarli catchment (42 km²) in the Central Spanish Pyrenees. The badlands have been identified as one of the main sediment sources for siltation of the downstream Barasona Reservoir. More than 700 tags were placed in different terrain units using three experimental setups, including lab experiments: (i) intensive feasibility tests ranging from laboratory flume experiments to tracer studies under natural channel and slope conditions to compare the transport of RFID tags relative to colored particles of the natural sediment; (ii) several transects across the badland to investigate sediment transfer characteristics on different morphological units (i.e. channel, rills, slopes); (iii) a raster of 99 RFID tags covering a slope flank with vegetated and unvegetated parts to reveal the influence of vegetation to erosion and transport processes. The detection of transported tags was carried out with a mobile antenna system to map the spatial distribution of tags after selected rainfall events and with two stationary antennas in channel cross-sections for time-continuous observation of tag passage. From the observations, we derived transport distances and velocities for a one year period. The transport behavior of the RFID tags was similar to the micrite limestone material of the badland. These results indicate the important role of terrain position and vegetation patches for erosion patterns, sediment transfer and the associated time scales along the catchment. We assess the potentials and limitations of the RFID technique as a direct tracer for assessing water and sediment connectivity along hillslopes to first order channels.

Using frequency tagging to quantify attentional deployment in a visual divided attention task.

PubMed

Toffanin, Paolo; de Jong, Ritske; Johnson, Addie; Martens, Sander

2009-06-01

Frequency tagging is an EEG method based on the quantification of the steady state visual evoked potential (SSVEP) elicited from stimuli which flicker with a distinctive frequency. Because the amplitude of the SSVEP is modulated by attention such that attended stimuli elicit higher SSVEP amplitudes than do ignored stimuli, the method has been used to investigate the neural mechanisms of spatial attention. However, up to now it has not been shown whether the amplitude of the SSVEP is sensitive to gradations of attention and there has been debate about whether attention effects on the SSVEP are dependent on the tagging frequency used. We thus compared attention effects on SSVEP across three attention conditions-focused, divided, and ignored-with six different tagging frequencies. Participants performed a visual detection task (respond to the digit 5 embedded in a stream of characters). Two stimulus streams, one to the left and one to the right of fixation, were displayed simultaneously, each with a background grey square whose hue was sine-modulated with one of the six tagging frequencies. At the beginning of each trial a cue indicated whether targets on the left, right, or both sides should be responded to. Accuracy was higher in the focused- than in the divided-attention condition. SSVEP amplitudes were greatest in the focused-attention condition, intermediate in the divided-attention condition, and smallest in the ignored-attention condition. The effect of attention on SSVEP amplitude did not depend on the tagging frequency used. Frequency tagging appears to be a flexible technique for studying attention.

Design considerations and tradeoffs for passive RFID tags

NASA Astrophysics Data System (ADS)

Hussien, Faisal A.; Turker, Didem Z.; Srinivasan, Rangakrishnan; Mobarak, Mohamed S.; Cortes, Fernando P.; Sanchez-Sinencio, Edgar

2005-06-01

Radio Frequency Identification (RFID) systems are widely used in a variety of tracking, security and tagging applications. Their operation in non line-of-sight environments makes them superior over similar devices such as barcode and infrared tags. RFID systems span a wide range of applications: medical history storage, dental prosthesis tracking, oil drilling pipe and concrete stress monitoring, toll ways services, animal tracking applications, etc. Passive RFID tags generate their power from the incoming signal; therefore, they do not require a power source. Accordingly, minimizing the power consumption and the implementation area are usually the main design considerations. This paper presents a complete analysis on designing a passive RFID tag. A system design methodology is introduced including the main issues and tradeoffs between different design parameters. The uplink modulation techniques used (ASK, PSK, FSK, and PWM) are illustrated showing how to choose the appropriate signaling scheme for a specific data rate, a certain distance of operation and a limited power consumption budget. An antenna system (transmitter and receiver) is proposed providing the maximum distance of operation with the transmitted power stated by FCC regulations. The backscatter modulation scheme used in the downlink is shown whether to be ASK-BM or PSK-BM and the differences between them are discussed. The key building blocks such as the charge pump, voltage reference, and the regulator used to generate the DC supply voltage from the incoming RF signal are discussed along with their design tradeoffs. A complete architecture for a passive RFID tag is provided as an example to illustrate the proposed RFID tag design methodology.

Behavior and movements of adult spring Chinook salmon (Oncorhynchus tshawytscha) in the Chehalis River Basin, southwestern Washington, 2015

USGS Publications Warehouse

Liedtke, Theresa L.; Zimmerman, Mara S.; Tomka, Ryan G.; Holt, Curt; Jennings, Lyle

2016-09-14

Recent interest in flood control and restoration strategies in the Chehalis River Basin has increased the need to understand the current status and ecology of spring Chinook salmon. Based on the extended period between freshwater entry and spawn timing, spring Chinook salmon have the longest exposure of all adult Chinook salmon life histories to the low-flow and high water temperature conditions that typically occur during summer. About 100 adult spring Chinook salmon were found dead in the Chehalis River in July and August 2009. Adult Chinook salmon are known to hold in cool-water refugia during warm summer months, but the extent to which spring Chinook salmon might use thermal refugia in the Chehalis River is unknown. The movements and temperature exposures of adult spring Chinook salmon following their return to the Chehalis River were investigated using radiotelemetry and transmitters equipped with temperature sensors, combined with water temperature monitoring throughout the basin. A total of 23 spring Chinook salmon were radio-tagged between April and early July 2015; 11 were captured and released in the main-stem Chehalis River, and 12 were captured and released in the South Fork Newaukum River. Tagged fish were monitored with a combination of fixed-site monitoring locations and regular mobile tracking, from freshwater entry through the spawning period.Water temperature and flow conditions in the main-stem Chehalis River during 2015 were atypical compared to historical averages. Mean monthly water temperatures between March and July 2015 were higher than any decade since 1960 and mean daily flows were 30–70 percent of the flows in previous years. Overall, 96 percent of the tagged fish were detected, with a mean of 62 d in the detection history of tagged fish. Of the 11 fish released in the main-stem Chehalis River, six fish (55 percent) moved upstream, either shortly after release (2–7 d, 50 percent), or following a short delay (12–18 d, 50 percent). One fish released in the main-stem Chehalis River remained near the release location for 64 d before moving upstream.The final fates for the seven fish that moved upstream in the main-stem Chehalis River included two fish with unknown fates, two fish with a fate of pre-spawn mortality, and three fish that were assigned a fate of spawner. Four (36 percent) of the radio-tagged Chinook salmon released in the main-stem Chehalis River showed limited movement from their release sites, and were assigned fates of unknown (one fish), pre-spawn mortality (one fish), and spit/mortality (2 fish). The 12 spring Chinook salmon released in the South Fork Newaukum River remained in the South Fork Newaukum River throughout the study period. Five (42 percent) of these fish were actively moving through the spawning period and were assigned a fate of spawner. Seven (58 percent) of these fish were detected for a period following release, but their detection histories ended prior to the spawning period. The fates assigned to these seven fish included two fish with spit/mortality fates and five fish with fates of pre-spawn mortality. Tagged fish in both the Chehalis River and the South Fork Newaukum River showed limited movements during the peak water temperatures in July and August, and were not frequently detected at sites where water temperatures were greater than 21 °C. Pre-spawn mortality due to predation or harvest may be an important factor in the Chehalis River Basin as it was the assigned fate for 27 percent of the fish released in the main-stem Chehalis River and 42 percent of the fish released in the South Fork Newaukum River.This study represents a substantial contribution to the understanding of spring Chinook salmon in the Chehalis River Basin. The water temperatures and flow conditions during the 2015 study period were not typical of the historical conditions in the basin and the numbers of tagged fish monitored was relatively low, so results should be interpreted with those cautions in mind.

Introducing keytagging, a novel technique for the protection of medical image-based tests.

PubMed

Rubio, Óscar J; Alesanco, Álvaro; García, José

2015-08-01

This paper introduces keytagging, a novel technique to protect medical image-based tests by implementing image authentication, integrity control and location of tampered areas, private captioning with role-based access control, traceability and copyright protection. It relies on the association of tags (binary data strings) to stable, semistable or volatile features of the image, whose access keys (called keytags) depend on both the image and the tag content. Unlike watermarking, this technique can associate information to the most stable features of the image without distortion. Thus, this method preserves the clinical content of the image without the need for assessment, prevents eavesdropping and collusion attacks, and obtains a substantial capacity-robustness tradeoff with simple operations. The evaluation of this technique, involving images of different sizes from various acquisition modalities and image modifications that are typical in the medical context, demonstrates that all the aforementioned security measures can be implemented simultaneously and that the algorithm presents good scalability. In addition to this, keytags can be protected with standard Cryptographic Message Syntax and the keytagging process can be easily combined with JPEG2000 compression since both share the same wavelet transform. This reduces the delays for associating keytags and retrieving the corresponding tags to implement the aforementioned measures to only ≃30 and ≃90ms respectively. As a result, keytags can be seamlessly integrated within DICOM, reducing delays and bandwidth when the image test is updated and shared in secure architectures where different users cooperate, e.g. physicians who interpret the test, clinicians caring for the patient and researchers. Copyright © 2015 Elsevier Inc. All rights reserved.

Typing Local Control and State Using Flow Analysis

NASA Astrophysics Data System (ADS)

Guha, Arjun; Saftoiu, Claudiu; Krishnamurthi, Shriram

Programs written in scripting languages employ idioms that confound conventional type systems. In this paper, we highlight one important set of related idioms: the use of local control and state to reason informally about types. To address these idioms, we formalize run-time tags and their relationship to types, and use these to present a novel strategy to integrate typing with flow analysis in a modular way. We demonstrate that in our separation of typing and flow analysis, each component remains conventional, their composition is simple, but the result can handle these idioms better than either one alone.

Precise terrestrial time: A means for improved ballistic missile guidance analysis

NASA Technical Reports Server (NTRS)

Ehrsam, E. E.; Cresswell, S. A.; Mckelvey, G. R.; Matthews, F. L.

1978-01-01

An approach developed to improve the ground instrumentation time tagging accuracy and adapted to support the Minuteman ICBM program is desired. The Timing Insertion Unit (TIU) technique produces a telemetry data time tagging resolution of one tenth of a microsecond, with a relative intersite accuracy after corrections and velocity data (range, azimuth, elevation and range rate) also used in missile guidance system analysis can be correlated to within ten microseconds of the telemetry guidance data. This requires precise timing synchronization between the metric and telemetry instrumentation sites. The timing synchronization can be achieved by using the radar automatic phasing system time correlation methods. Other time correlation techniques such as Television (TV) Line-10 and the Geostationary Operational Environmental Satellites (GEOS) terrestial timing receivers are also considered.

An Identification Procedure for Behavioral Analysis in a Multi-User environment.

PubMed

Guerra, Claudio; Bianchi, Valentina; De Munari, Ilaria; Ciampolini, Paolo

2015-01-01

As the average age of the EU population increases, ICT solutions are going to play a key role in order to find answers to the new challenges the demographic change is carrying on. At the University of Parma an AAL (Ambient Assisted Living) system named CARDEA has been developed during the last 10 years. Within CARDEA, behavioral analysis is carried out, based on environmental sensors. If multiple users live in the same environment, however, data coming from sensors need to be properly tagged: in this paper, a simple technique for such tagging is proposed, which exploits the same wireless transmission used for transmitting data, thus not requiring additional hardware components and avoiding more complex and expensive (radio)localization techniques. Preliminary results are shown, featuring a satisfactory accuracy.

Opo lidar sounding of trace atmospheric gases in the 3 - 4 μm spectral range

NASA Astrophysics Data System (ADS)

Romanovskii, Oleg A.; Sadovnikov, Sergey A.; Kharchenko, Olga V.; Yakovlev, Semen V.

2018-04-01

The applicability of a KTA crystal-based laser system with optical parametric oscillators (OPO) generation to lidar sounding of the atmosphere in the spectral range 3-4 μm is studied in this work. A technique developed for lidar sounding of trace atmospheric gases (TAG) is based on differential absorption lidar (DIAL) method and differential optical absorption spectroscopy (DOAS). The DIAL-DOAS technique is tested to estimate its efficiency for lidar sounding of atmospheric trace gases. The numerical simulation performed shows that a KTA-based OPO laser is a promising source of radiation for remote DIAL-DOAS sounding of the TAGs under study along surface tropospheric paths. A possibility of using a PD38-03-PR photodiode for the DIAL gas analysis of the atmosphere is shown.

Optimal MEMS device for mobility and zeta potential measurements using DC electrophoresis.

PubMed

Karam, Pascal R; Dukhin, Andrei; Pennathur, Sumita

2017-05-01

We have developed a novel microchannel geometry that allows us to perform simple DC electrophoresis to measure the electrophoretic mobility and zeta potential of analytes and particles. In standard capillary geometries, mobility measurements using DC fields are difficult to perform. Specifically, measurements in open capillaries require knowledge of the hard to measure and often dynamic wall surface potential. Although measurements in closed capillaries eliminate this requirement, the measurements must be performed at infinitesimally small regions of zero flow where the pressure driven-flow completely cancels the electroosmotic flow (Komagata Planes). Furthermore, applied DC fields lead to electrode polarization, further questioning the reliability and accuracy of the measurement. In contrast, our geometry expands and moves the Komagata planes to where velocity gradients are at a minimum, and thus knowledge of the precise location of a Komagata plane is not necessary. Additionally, our microfluidic device prevents electrode polarization because of fluid recirculation around the electrodes. We fabricated our device using standard MEMS fabrication techniques and performed electrophoretic mobility measurements on 500 nm fluorescently tagged polystyrene particles at various buffer concentrations. Results are comparable to two different commercial dynamic light scattering based particle sizing instruments. We conclude with guidelines to further develop this robust electrophoretic tool that allows for facile and efficient particle characterization. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A Semantic Rule-Based Framework for Efficient Retrieval of Educational Materials

ERIC Educational Resources Information Center

Mahmoudi, Maryam Tayefeh; Taghiyareh, Fattaneh; Badie, Kambiz

2013-01-01

Retrieving resources in an appropriate manner has a promising role in increasing the performance of educational support systems. A variety of works have been done to organize materials for educational purposes using tagging techniques. Despite the effectiveness of these techniques within certain domains, organizing resources in a way being…

Exploring Ways to Improve DTAG Deployment Success Rates With the ARTS Pneumatic Launcher

DTIC Science & Technology

2010-01-01

absorber system has been developed, (the giraffe leg technique or GL, Figure 2), which has the shock absorbing component in front of the tag. The ARTS...3 Figure 2: the new 1- and 4-spring shock absorbing robot (left) and the GL ( giraffe leg technique) with 3 studs (right) WORK

Rapid purification of fluorescent enzymes by ultrafiltration

NASA Technical Reports Server (NTRS)

Benjaminson, M. A.; Satyanarayana, T.

1983-01-01

In order to expedite the preparation of fluorescently tagged enzymes for histo-cyctochemistry, a previously developed method employing gel column purification was compared with a more rapid modern technique using the Millipore Immersible CX-ultrafilter. Microscopic evaluation of the resulting conjugates showed comparable products. Much time and effort is saved using the new technique.

Rapid purification of fluorescent enzymes by ultrafiltration

NASA Technical Reports Server (NTRS)

Benjaminson, M. A.; Satyanarayana, T.

1983-01-01

In order to expedite the preparation of fluorescently tagged enzymes for histo/cytochemistry, a previously developed method employing gel column purification was compared with a more rapid modern technique using the Millipore Immersible CX-ultrafilter. Microscopic evaluation of the resulting conjugates showed comparable products. Much time and effort is saved using the new technique.

Advantage of spatial map ion imaging in the study of large molecule photodissociation

NASA Astrophysics Data System (ADS)

Lee, Chin; Lin, Yen-Cheng; Lee, Shih-Huang; Lee, Yin-Yu; Tseng, Chien-Ming; Lee, Yuan-Tseh; Ni, Chi-Kung

2017-07-01

The original ion imaging technique has low velocity resolution, and currently, photodissociation is mostly investigated using velocity map ion imaging. However, separating signals from the background (resulting from undissociated excited parent molecules) is difficult when velocity map ion imaging is used for the photodissociation of large molecules (number of atoms ≥ 10). In this study, we used the photodissociation of phenol at the S1 band origin as an example to demonstrate how our multimass ion imaging technique, based on modified spatial map ion imaging, can overcome this difficulty. The photofragment translational energy distribution obtained when multimass ion imaging was used differed considerably from that obtained when velocity map ion imaging and Rydberg atom tagging were used. We used conventional translational spectroscopy as a second method to further confirm the experimental results, and we conclude that data should be interpreted carefully when velocity map ion imaging or Rydberg atom tagging is used in the photodissociation of large molecules. Finally, we propose a modified velocity map ion imaging technique without the disadvantages of the current velocity map ion imaging technique.

Detection, Identification, Location, and Remote Sensing Using SAW RFID Sensor Tags

NASA Technical Reports Server (NTRS)

Barton, Richard J.; Kennedy, Timothy F.; Williams, Robert M.; Fink, Patrick W.; Ngo, Phong H.

2009-01-01

The Electromagnetic Systems Branch (EV4) of the Avionic Systems Division at NASA Johnson Space Center in Houston, TX is studying the utility of surface acoustic wave (SAW) radiofrequency identification (RFID) tags for multiple wireless applications including detection, identification, tracking, and remote sensing of objects on the lunar surface, monitoring of environmental test facilities, structural shape and health monitoring, and nondestructive test and evaluation of assets. For all of these applications, it is anticipated that the system utilized to interrogate the SAW RFID tags may need to operate at fairly long range and in the presence of considerable multipath and multiple-access interference. Towards that end, EV4 is developing a prototype SAW RFID wireless interrogation system for use in such environments called the Passive Adaptive RFID Sensor Equipment (PARSED) system. The system utilizes a digitally beam-formed planar receiving antenna array to extend range and provide direction-of-arrival information coupled with an approximate maximum-likelihood signal processing algorithm to provide near-optimal estimation of both range and temperature. The system is capable of forming a large number of beams within the field of view and resolving the information from several tags within each beam. The combination of both spatial and waveform discrimination provides the capability to track and monitor telemetry from a large number of objects appearing simultaneously within the field of view of the receiving array. In this paper, we will consider the application of the PARSEQ system to the problem of simultaneous detection, identification, localization, and temperature estimation for multiple objects. We will summarize the overall design of the PARSEQ system and present a detailed description of the design and performance of the signal detection and estimation algorithms incorporated in the system. The system is currently configured only to measure temperature (jointly with range and tag ID), but future versions will be revised to measure parameters other than temperature as SAW tags capable of interfacing with external sensors become available. It is anticipated that the estimation of arbitrary parameters measured using SAW-based sensors will be based on techniques very similar to the joint range and temperature estimation techniques described in this paper.

Nano-technology and privacy: on continuous surveillance outside the panopticon.

PubMed

Hoven, Jeroen Van Den; Vermaas, Pieter E

2007-01-01

We argue that nano-technology in the form of invisible tags, sensors, and Radio Frequency Identity Chips (RFIDs) will give rise to privacy issues that are in two ways different from the traditional privacy issues of the last decades. One, they will not exclusively revolve around the idea of centralization of surveillance and concentration of power, as the metaphor of the Panopticon suggests, but will be about constant observation at decentralized levels. Two, privacy concerns may not exclusively be about constraining information flows but also about designing of materials and nano-artifacts such as chips and tags. We begin by presenting a framework for structuring the current debates on privacy, and then present our arguments.

An introduction to mass cytometry: fundamentals and applications.

PubMed

Tanner, Scott D; Baranov, Vladimir I; Ornatsky, Olga I; Bandura, Dmitry R; George, Thaddeus C

2013-05-01

Mass cytometry addresses the analytical challenges of polychromatic flow cytometry by using metal atoms as tags rather than fluorophores and atomic mass spectrometry as the detector rather than photon optics. The many available enriched stable isotopes of the transition elements can provide up to 100 distinguishable reporting tags, which can be measured simultaneously because of the essential independence of detection provided by the mass spectrometer. We discuss the adaptation of traditional inductively coupled plasma mass spectrometry to cytometry applications. We focus on the generation of cytometry-compatible data and on approaches to unsupervised multivariate clustering analysis. Finally, we provide a high-level review of some recent benchmark reports that highlight the potential for massively multi-parameter mass cytometry.

CPTAC Evaluates Long-Term Reproducibility of Quantitative Proteomics Using Breast Cancer Xenografts | Office of Cancer Clinical Proteomics Research

Cancer.gov

Liquid chromatography tandem-mass spectrometry (LC-MS/MS)- based methods such as isobaric tags for relative and absolute quantification (iTRAQ) and tandem mass tags (TMT) have been shown to provide overall better quantification accuracy and reproducibility over other LC-MS/MS techniques. However, large scale projects like the Clinical Proteomic Tumor Analysis Consortium (CPTAC) require comparisons across many genomically characterized clinical specimens in a single study and often exceed the capability of traditional iTRAQ-based quantification.

Resetting the T Cell Repertoire in Prostate Cancer Bearing Host

DTIC Science & Technology

2009-03-01

splenocytes were harvested and stained with fluorochrome conjugated antibodies anti CD4 (RM4.5), anti CDS (53 6.7), and anti V(3S.I+S.2 (MRS 2; BD... harvested 20 h later and analyzed by flow cytome~ for the relative abundance of CFSEiow (SV40 TAg peptide) and CFSE i (HSV peptide) populations. Detection...and spleens were harvested S days after the last injection. Cells were stained for flow cytometry analysis. This treatment did not affect either

SU-E-I-65: Estimation of Tagging Efficiency in Pseudo-Continuous Arterial Spin Labeling (pCASL) MRI

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

Jen, M; Yan, F; Tseng, Y

2015-06-15

Purpose: pCASL was recommended as a potent approach for absolute cerebral blood flow (CBF) quantification in clinical practice. However, uncertainties of tagging efficiency in pCASL remain an issue. This study aimed to estimate tagging efficiency by using short quantitative pulsed ASL scan (FAIR-QUIPSSII) and compare resultant CBF values with those calibrated by using 2D Phase Contrast (PC) MRI. Methods: Fourteen normal volunteers participated in this study. All images, including whole brain (WB) pCASL, WB FAIR-QUIPSSII and single-slice 2D PC, were collected on a 3T clinical MRI scanner with a 8-channel head coil. DeltaM map was calculated by averaging the subtractionmore » of tag/control pairs in pCASL and FAIR-QUIPSSII images and used for CBF calculation. Tagging efficiency was then calculated by the ratio of mean gray matter CBF obtained from pCASL and FAIR-QUIPSSII. For comparison, tagging efficiency was also estimated with 2D PC, a previously established method, by contrast WB CBF in pCASL and 2D PC. Feasibility of estimation from a short FAIR-QUIPSSII scan was evaluated by number of averages required for obtaining a stable deltaM value. Setting deltaM calculated by maximum number of averaging (50 pairs) as reference, stable results were defined within ±10% variation. Results: Tagging efficiencies obtained by 2D PC MRI (0.732±0.092) were significantly lower than which obtained by FAIRQUIPPSSII (0.846±0.097) (P<0.05). Feasibility results revealed that four pairs of images in FAIR-QUIPPSSII scan were sufficient to obtain a robust calibration of less than 10% differences from using 50 pairs. Conclusion: This study found that reliable estimation of tagging efficiency could be obtained by a few pairs of FAIR-QUIPSSII images, which suggested that calibration scan in a short duration (within 30s) was feasible. Considering recent reports concerning variability of PC MRI-based calibration, this study proposed an effective alternative for CBF quantification with pCASL.« less

Development of an instrument for non-destructive identification of Unexploded Ordnance using tagged neutrons - a proof of concept study

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

Mitra, S.; Dioszegi, I.

2011-10-23

Range clearance operations at munitions testing grounds must discriminate Unexploded Ordnance (UXO) from clutter items and distinguish UXO filled with High Explosives (HE) from those with inert fillers. Non-destructive technologies are thus necessary for the cost-effective disposal of UXO during remediation of such sites. The only technique showing promise so far for the non-destructive elemental characterization of UXO fillers utilizes neutron interactions with the material to detect carbon (C), nitrogen (N) and oxygen (O) which have unique ratios in HE. However, several unresolved issues hinder the wide application of this potentially very suitable technique. The most important one is thatmore » neutrons interact with all surrounding matter in addition to the interrogated material, leading to a very high gamma-ray background in the detector. Systems requiring bulky shielding and having poor signal-to-noise ratios (SNRs) for measuring elements are unsuitable for field deployment. The inadequacies of conventional neutron interrogation methods are overcome by using the tagged-neutron approach, and the availability of compact sealed neutron generators exploiting this technique offers field deployment of non-intrusive measurement systems for detecting threat materials, like explosives and drugs. By accelerating deuterium ions into a tritium target, the subsequent fusion reaction generates nearly back-to-back emissions of neutrons and alpha particles of energy 14.1 and 3.5 MeV respectively. A position-sensitive detector recognizes the associated alpha particle, thus furnishing the direction of the neutron. The tagged neutrons interact with the nuclei of the interrogated object, producing element-specific prompt gamma-rays that the gamma detectors recognize. Measuring the delay between the detections of the alpha particle and the gamma-ray determines where the reaction occurred along the axis of the neutron beam (14.1 MeV neutrons travel at 5 cm/nanosecond, while gamma rays cover 30 cm/nanosecond). The main advantage of the technique is its ability to simultaneously provide 2D and 3D imaging of objects and their elemental composition. This work reports on the efficacy of using 14 MeV neutrons tagged by the associated particle neutron time-of-flight technique (APnTOF) to extract neutron induced characteristic gamma-rays from an object-of-interest with high SNR and without interference from nearby clutter.« less

Gene doctoring: a method for recombineering in laboratory and pathogenic Escherichia coli strains.

PubMed

Lee, David J; Bingle, Lewis E H; Heurlier, Karin; Pallen, Mark J; Penn, Charles W; Busby, Stephen J W; Hobman, Jon L

2009-12-09

Homologous recombination mediated by the lambda-Red genes is a common method for making chromosomal modifications in Escherichia coli. Several protocols have been developed that differ in the mechanisms by which DNA, carrying regions homologous to the chromosome, are delivered into the cell. A common technique is to electroporate linear DNA fragments into cells. Alternatively, DNA fragments are generated in vivo by digestion of a donor plasmid with a nuclease that does not cleave the host genome. In both cases the lambda-Red gene products recombine homologous regions carried on the linear DNA fragments with the chromosome. We have successfully used both techniques to generate chromosomal mutations in E. coli K-12 strains. However, we have had limited success with these lambda-Red based recombination techniques in pathogenic E. coli strains, which has led us to develop an enhanced protocol for recombineering in such strains. Our goal was to develop a high-throughput recombineering system, primarily for the coupling of genes to epitope tags, which could also be used for deletion of genes in both pathogenic and K-12 E. coli strains. To that end we have designed a series of donor plasmids for use with the lambda-Red recombination system, which when cleaved in vivo by the I-SceI meganuclease generate a discrete linear DNA fragment, allowing for C-terminal tagging of chromosomal genes with a 6xHis, 3xFLAG, 4xProteinA or GFP tag or for the deletion of chromosomal regions. We have enhanced existing protocols and technologies by inclusion of a cassette conferring kanamycin resistance and, crucially, by including the sacB gene on the donor plasmid, so that all but true recombinants are counter-selected on kanamycin and sucrose containing media, thus eliminating the need for extensive screening. This method has the added advantage of limiting the exposure of cells to the potential damaging effects of the lambda-Red system, which can lead to unwanted secondary alterations to the chromosome. We have developed a counter-selective recombineering technique for epitope tagging or for deleting genes in E. coli. We have demonstrated the versatility of the technique by modifying the chromosome of the enterohaemorrhagic O157:H7 (EHEC), uropathogenic CFT073 (UPEC), enteroaggregative O42 (EAEC) and enterotoxigenic H10407 (ETEC) E. coli strains as well as in K-12 laboratory strains.

Barium Tagging for nEXO

NASA Astrophysics Data System (ADS)

Fudenberg, Daniel; Brunner, Thomas; Varentsov, Victor; Devoe, Ralph; Dilling, Jens; Gratta, Giorgio; nEXO Collaboration

2015-10-01

nEXO is a next-generation experiment designed to search for 0 νββ -decay of Xe-136 in a liquid xenon time projection chamber. Positive observation of this decay would determine the neutrino to be a Majorana particle In order to greatly reduce background contributions to this search, the collaboration is developing several ``barium tagging'' techniques to recover and identify the decay daughter, Ba-136. ``Tagging'' may be available for a 2nd phase of nEXO and will push the sensitivity beyond the inverted neutrino-mass hierarchy. Tagging methods in testing for this phase include Ba-ion capture on a probe with identification by resonance ionization laser spectroscopy, and Ba capture in solid xenon on a cold probe with identification by fluorescence. In addition, Ba tagging for a gas-phase detector, appropriate for a later stage, is being tested. Here efficient ion extraction from heavy carrier gases is key. Detailed gas-dynamic and ion transport calculations have been performed to optimize for ion extraction. An apparatus to extract Ba ions from up to 10 bar xenon gas into vacuum using an RF-only funnel has been constructed and demonstrates extraction of ions from noble gases. We will present this system's status along with results of this R&D program.

The Monitoring and Affinity Purification of Proteins Using Dual Tags with Tetracysteine Motifs

NASA Astrophysics Data System (ADS)

Giannone, Richard J.; Liu, Yie; Wang, Yisong

Identification and characterization of protein-protein interaction networks is essential for the elucidation of biochemical mechanisms and cellular function. Affinity purification in combination with liquid chromatography-tandem mass spectrometry (LC-MS/MS) has emerged as a very powerful tactic for the identification of specific protein-protein interactions. In this chapter, we describe a comprehensive methodology that uses our recently developed dual-tag affinity purification system for the enrichment and identification of mammalian protein complexes. The protocol covers a series of separate but sequentially related techniques focused on the facile monitoring and purification of a dual-tagged protein of interest and its interacting partners via a system built with tetracysteine motifs and various combinations of affinity tags. Using human telomeric repeat binding factor 2 (TRF2) as an example, we demonstrate the power of the system in terms of bait protein recovery after dual-tag affinity purification, detection of bait protein subcellular localization and expression, and successful identification of known and potentially novel TRF2 interacting proteins. Although the protocol described here has been optimized for the identification and characterization of TRF2-associated proteins, it is, in principle, applicable to the study of any other mammalian protein complexes that may be of interest to the research community.

Distinguishing Aspartic and Isoaspartic Acids in Peptides by Several Mass Spectrometric Fragmentation Methods.

PubMed

DeGraan-Weber, Nick; Zhang, Jun; Reilly, James P

2016-12-01

Six ion fragmentation techniques that can distinguish aspartic acid from its isomer, isoaspartic acid, were compared. MALDI post-source decay (PSD), MALDI 157 nm photodissociation, tris(2,4,6-trimethoxyphenyl)phosphonium bromide (TMPP) charge tagging in PSD and photodissociation, ESI collision-induced dissociation (CID), electron transfer dissociation (ETD), and free-radical initiated peptide sequencing (FRIPS) with CID were applied to peptides containing either aspartic or isoaspartic acid. Diagnostic ions, such as the y-46 and b+H 2 O, are present in PSD, photodissociation, and charge tagging. c • +57 and z-57 ions are observed in ETD and FRIPS experiments. For some molecules, aspartic and isoaspartic acid yield ion fragments with significantly different intensities. ETD and charge tagging appear to be most effective at distinguishing these residues. Graphical Abstract ᅟ.

Measurement of the neutral B meson-antineutral B meson oscillation frequency using fully-reconstructed semileptonic decays

NASA Astrophysics Data System (ADS)

Leclerc, Christopher Mark

B0B¯0 flavor oscillation frequency is studied by examining events containing a fully reconstructed B → D*lnu. The opposite side flavor is determined with tagging tools that look for characteristic kaon and lepton tags, plus a neural network output for events with conflicting or non-existent kaon/lepton tags. Based on a data set of 20.6 fb -1 on the Upsilon(4s) resonance, 2.6 fb -1 offpeak, Deltamd = 0.492 +/- 0.018 +/- 0.013 ps-1. The first error is statistical and the second is systematic. The B 0 lifetime is also measured, tB0=1.523+0.024- 0.023+/-0.022ps . The Deltamd measurement is competitive with other recent measurements and develops several novel and powerful data analysis techniques.

FRET-based binding assay between a fluorescent cAMP analogue and a cyclic nucleotide-binding domain tagged with a CFP.

PubMed

Romero, Francisco; Santana-Calvo, Carmen; Sánchez-Guevara, Yoloxochitl; Nishigaki, Takuya

2017-09-01

The cyclic nucleotide-binding domain (CNBD) functions as a regulatory domain of many proteins involved in cyclic nucleotide signalling. We developed a straightforward and reliable binding assay based on intermolecular fluorescence resonance energy transfer (FRET) between an adenosine-3', 5'-cyclic monophosphate analogue labelled with fluorescein and a recombinant CNBD of human EPAC1 tagged with a cyan fluorescence protein (CFP). The high FRET efficiency of this method (~ 80%) allowed us to perform several types of binding experiments with nanomolar range of sample using conventional equipment. In addition, the CFP tag on the CNBD enabled us to perform a specific binding experiment using an unpurified protein. Considering these advantages, this technique is useful to study poorly characterized CNBDs. © 2017 Federation of European Biochemical Societies.

On-site detection of stacked genetically modified soybean based on event-specific TM-LAMP and a DNAzyme-lateral flow biosensor.

PubMed

Cheng, Nan; Shang, Ying; Xu, Yuancong; Zhang, Li; Luo, Yunbo; Huang, Kunlun; Xu, Wentao

2017-05-15

Stacked genetically modified organisms (GMO) are becoming popular for their enhanced production efficiency and improved functional properties, and on-site detection of stacked GMO is an urgent challenge to be solved. In this study, we developed a cascade system combining event-specific tag-labeled multiplex LAMP with a DNAzyme-lateral flow biosensor for reliable detection of stacked events (DP305423× GTS 40-3-2). Three primer sets, both event-specific and soybean species-specific, were newly designed for the tag-labeled multiplex LAMP system. A trident-like lateral flow biosensor displayed amplified products simultaneously without cross contamination, and DNAzyme enhancement improved the sensitivity effectively. After optimization, the limit of detection was approximately 0.1% (w/w) for stacked GM soybean, which is sensitive enough to detect genetically modified content up to a threshold value established by several countries for regulatory compliance. The entire detection process could be shortened to 120min without any large-scale instrumentation. This method may be useful for the in-field detection of DP305423× GTS 40-3-2 soybean on a single kernel basis and on-site screening tests of stacked GM soybean lines and individual parent GM soybean lines in highly processed foods. Copyright © 2017 Elsevier B.V. All rights reserved.

Noncontrast-enhanced renal angiography using multiple inversion recovery and alternating TR balanced steady-state free precession.

PubMed

Dong, Hattie Z; Worters, Pauline W; Wu, Holden H; Ingle, R Reeve; Vasanawala, Shreyas S; Nishimura, Dwight G

2013-08-01

Noncontrast-enhanced renal angiography techniques based on balanced steady-state free precession avoid external contrast agents, take advantage of high inherent blood signal from the T 2 / T 1 contrast mechanism, and have short steady-state free precession acquisition times. However, background suppression is limited; inflow times are inflexible; labeling region is difficult to define when tagging arterial flow; and scan times are long. To overcome these limitations, we propose the use of multiple inversion recovery preparatory pulses combined with alternating pulse repetition time balanced steady-state free precession to produce renal angiograms. Multiple inversion recovery uses selective spatial saturation followed by four nonselective inversion recovery pulses to concurrently null a wide range of background T 1 species while allowing for adjustable inflow times; alternating pulse repetition time steady-state free precession maintains vessel contrast and provides added fat suppression. The high level of suppression enables imaging in three-dimensional as well as projective two-dimensional formats, the latter of which has a scan time as short as one heartbeat. In vivo studies at 1.5 T demonstrate the superior vessel contrast of this technique. © 2012 Wiley Periodicals, Inc.

Optimal Suturing Technique and Number of Sutures for Surgical Implantation of Acoustic Transmitters in Juvenile Salmonids

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

Deters, Katherine A.; Brown, Richard S.; Boyd, James W.

2012-01-02

The size reduction of acoustic transmitters has led to a reduction in the length of incision needed to implant a transmitter. Smaller suture knot profiles and fewer sutures may be adequate for closing an incision used to surgically implant an acoustic microtransmitter. As a result, faster surgery times and reduced tissue trauma could lead to increased survival and decreased infection for implanted fish. The objective of this study was to assess the effects of five suturing techniques on mortality, tag and suture retention, incision openness, ulceration, and redness in juvenile Chinook salmon Oncorhynchus tshawytscha implanted with acoustic microtransmitters. Suturing wasmore » performed by three surgeons, and study fish were held at two water temperatures (12°C and 17°C). Mortality was low and tag retention was high for all treatments on all examination days (7, 14, 21, and 28 days post-surgery). Because there was surgeon variation in suture retention among treatments, further analyses included only the one surgeon who received feedback training in all suturing techniques. Incision openness and tissue redness did not differ among treatments. The only difference observed among treatments was in tissue ulceration. Incisions closed with a horizontal mattress pattern had more ulceration than other treatments among fish held for 28 days at 17°C. Results from this study suggest that one simple interrupted 1 × 1 × 1 × 1 suture is adequate for closing incisions on fish under most circumstances. However, in dynamic environments, two simple interrupted 1 × 1 × 1 × 1 sutures should provide adequate incision closure. Reducing bias in survival and behavior tagging studies is important when making comparisons to the migrating salmon population. Therefore, by minimizing the effects of tagging on juvenile salmon (reduced tissue trauma and reduced surgery time), researchers can more accurately estimate survival and behavior.« less

Pulmonary Edema and Plasma Volume Changes in Dysbarism. M.S. Thesis - Texas Univ.

NASA Technical Reports Server (NTRS)

Joki, J. A.

1972-01-01

Two groups of anesthetized, fasted pigs were utilized. One group of 13 animals (8.5 to 16.6 kilograms) was exposed to a high-pressure environment, and the other group of eight animals (6.9 to 20.0 kilograms) constituted the control group. The experimental group was subjected to an atmosphere of 90 percent nitrogen and 10 percent oxygen at a pressure of 50 psig for 30 minutes and then decompressed at a rate 10 psi/min. Plasma volumes, using both iodine-125-tagged-albumin and chromium-51-tagged-cell dilution techniques, were measured before, immediately after, and at 30 and 60 minutes after decompression. Aortic and right-ventricular systolic pressures were also recorded. At 60 minutes after decompression, blood samples were taken, the animals were sacrificed, and the water content of the lungs, kidneys, livers, and spleens was estimated by measuring tissue wet weight and dry weight. Protein extravasation and tissue blood volumes were determined by measuring the iodine-125-tagged-albumin and chromium-51-tagged-cell spaces in homo-genates of the organs under investigation.

Selection of mRNA 5'-untranslated region sequence with high translation efficiency through ribosome display

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

Mie, Masayasu; Shimizu, Shun; Takahashi, Fumio

2008-08-15

The 5'-untranslated region (5'-UTR) of mRNAs functions as a translation enhancer, promoting translation efficiency. Many in vitro translation systems exhibit a reduced efficiency in protein translation due to decreased translation initiation. The use of a 5'-UTR sequence with high translation efficiency greatly enhances protein production in these systems. In this study, we have developed an in vitro selection system that favors 5'-UTRs with high translation efficiency using a ribosome display technique. A 5'-UTR random library, comprised of 5'-UTRs tagged with a His-tag and Renilla luciferase (R-luc) fusion, were in vitro translated in rabbit reticulocytes. By limiting the translation period, onlymore » mRNAs with high translation efficiency were translated. During translation, mRNA, ribosome and translated R-luc with His-tag formed ternary complexes. They were collected with translated His-tag using Ni-particles. Extracted mRNA from ternary complex was amplified using RT-PCR and sequenced. Finally, 5'-UTR with high translation efficiency was obtained from random 5'-UTR library.« less

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

Nekoogar, F; Dowla, F

An IAEA Technical Meeting on Techniques for IAEA Verification of Enrichment Activities identified 'smart tags' as a technology that should be assessed for tracking and locating UF6 cylinders. Although there is vast commercial industry working on RFID systems, the vulnerabilities of commercial products are only beginning to emerge. Most of the commercially off-the-shelf (COTS) RFID systems operate in very narrow frequency bands, making them vulnerable to detection, jamming and tampering and also presenting difficulties when used around metals (i.e. UF6 cylinders). Commercial passive RFID tags have short range, while active RFID tags that provide long ranges have limited lifetimes. Theremore » are also some concerns with the introduction of strong (narrowband) radio frequency signals around radioactive and nuclear materials. Considering the shortcomings of commercial RFID systems, in their current form, they do not offer a promising solution for continuous monitoring and tracking of UF6 cylinders. In this paper, we identify the key challenges faced by commercial RFID systems for monitoring UF6 cylinders, and introduce an ultra-wideband approach for tag/reader communications that addresses most of the identified challenges for IAEA safeguards applications.« less

Characterization and Determination of Interesterification Markers (Triacylglycerol Regioisomers) in Confectionery Oils by Liquid Chromatography-Mass Spectrometry

PubMed Central

Santoro, Valentina; Aigotti, Riccardo; Gastaldi, Daniela; Romaniello, Francesco; Forte, Emanuele; Magni, Martina; Baiocchi, Claudio

2018-01-01

Interesterification is an industrial transformation process aiming to change the physico-chemical properties of vegetable oils by redistributing fatty acid position within the original constituent of the triglycerides. In the confectionery industry, controlling formation degree of positional isomers is important in order to obtain fats with the desired properties. Silver ion HPLC (High Performance Liquid Chromatography) is the analytical technique usually adopted to separate triglycerides (TAGs) having different unsaturation degrees. However, separation of TAG positional isomers is a challenge when the number of double bonds is the same and the only difference is in their position within the triglyceride molecule. The TAG positional isomers involved in the present work have a structural specificity that require a separation method tailored to the needs of confectionery industry. The aim of this work was to obtain a chromatographic resolution that might allow reliable qualitative and quantitative evaluation of TAG positional isomers within reasonably rapid retention times and robust in respect of repeatability and reproducibility. The resulting analytical procedure was applied both to confectionery raw materials and final products. PMID:29462917

From Trioleoyl glycerol to extra virgin olive oil through multicomponent triacylglycerol mixtures: Crystallization and polymorphic transformation examined with differential scanning calorimetry and X-ray diffration techniques.

PubMed

Bayés-García, L; Calvet, T; Cuevas-Diarte, M A; Ueno, S

2017-09-01

The polymorphic crystallization and transformation behavior of extra virgin olive oil (EVOO) was examined by using differential scanning calorimetry (DSC) and X-ray diffraction with both laboratory-scale (XRD) and synchrotron radiation source (SR-XRD). The complex behavior observed was studied by previously analyzing mixtures composed by its main 2 to 6 triacylglycerol (TAG) components. Thus, component TAGs were successively added to simulate EVOO composition, until reaching a 6 TAGs mixture, composed by trioleoyl glycerol (OOO), 1-palmitoyl-2,3-dioleoyl glycerol (POO), 1,2-dioleoyl-3-linoleoyl glycerol (OOL), 1-palmitoyl-2-oleoyl-3-linoleoyl glycerol (POL), 1,2-dipalmitoyl-3-oleoyl glycerol (PPO) and 1-stearoyl-2,3-dioleoyl glycerol (SOO). Molten samples were cooled from 25°C to -80°C at a controlled rate of 2°C/min and subsequently heated at the same rate. The polymorphic behavior observed in multicomponent TAG mixtures was interpreted by considering three main groups of TAGs with different molecular structures: triunsaturated OOO and OOL, saturated-unsaturated-unsaturated POO, POL and SOO, and saturated-saturated-unsaturated PPO. As confirmed by our previous work, TAGs belonging to the same structural group displayed a highly similar polymorphic behavior. EVOO exhibited two different β'-2L polymorphic forms (β' 2 -2L and β' 1 -2L), which transformed into β'-3L when heated. Equivalent polymorphic pathways were detected when the same experimental conditions were applied to the 6 TAG components mixture. Hence, minor components may not exert a strong influence in this case. Copyright © 2017 Elsevier Ltd. All rights reserved.

Insight into the coordination and the binding sites of Cu(2+) by the histidyl-6-tag using experimental and computational tools.

PubMed

Watly, Joanna; Simonovsky, Eyal; Wieczorek, Robert; Barbosa, Nuno; Miller, Yifat; Kozlowski, Henryk

2014-07-07

His-tags are specific sequences containing six to nine subsequent histydyl residues, and they are used for purification of recombinant proteins by use of IMAC chromatography. Such polyhistydyl tags, often used in molecular biology, can be also found in nature. Proteins containing histidine-rich domains play a critical role in many life functions in both prokaryote and eukaryote organisms. Binding mode and the thermodynamic properties of the system depend on the specific metal ion and the histidine sequence. Despite the wide application of the His-tag for purification of proteins, little is known about the properties of metal-binding to such tag domains. This inspired us to undertake detailed studies on the coordination of Cu(2+) ion to hexa-His-tag. Experiments were performed using the potentiometric, UV-visible, CD, and EPR techniques. In addition, molecular dynamics (MD) simulations and density functional theory (DFT) calculations were applied. The experimental studies have shown that the Cu(2+) ion binds most likely to two imidazoles and one, two, or three amide nitrogens, depending on the pH. The structures and stabilities of the complexes for the Cu(2+)-Ac-(His)6-NH2 system using experimental and computational tools were established. Polymorphic binding states are suggested, with a possibility of the formation of α-helix structure induced by metal ion coordination. Metal ion is bound to various pairs of imidazole moieties derived from the tag with different efficiencies. The coordination sphere around the metal ion is completed by molecules of water. Finally, the Cu(2+) binding by Ac-(His)6-NH2 is much more efficient compared to other multihistidine protein domains.

Discovering Indicators of Successful Collaboration Using Tense: Automated Extraction of Patterns in Discourse

ERIC Educational Resources Information Center

Thompson, Kate; Kennedy-Clark, Shannon; Wheeler, Penny; Kelly, Nick

2014-01-01

This paper describes a technique for locating indicators of success within the data collected from complex learning environments, proposing an application of e-research to access learner processes and measure and track group progress. The technique combines automated extraction of tense and modality via parts-of-speech tagging with a visualisation…

Smart Collections: Can Artificial Intelligence Tools and Techniques Assist with Discovering, Evaluating and Tagging Digital Learning Resources?

ERIC Educational Resources Information Center

Leibbrandt, Richard; Yang, Dongqiang; Pfitzner, Darius; Powers, David; Mitchell, Pru; Hayman, Sarah; Eddy, Helen

2010-01-01

This paper reports on a joint proof of concept project undertaken by researchers from the Flinders University Artificial Intelligence Laboratory in partnership with information managers from the Education Network Australia (edna) team at Education Services Australia to address the question of whether artificial intelligence techniques could be…

Helium Tagging Infrared Photodissociation Spectroscopy of Reactive Ions.

PubMed

Roithová, Jana; Gray, Andrew; Andris, Erik; Jašík, Juraj; Gerlich, Dieter

2016-02-16

The interrogation of reaction intermediates is key for understanding chemical reactions; however their direct observation and study remains a considerable challenge. Mass spectrometry is one of the most sensitive analytical techniques, and its use to study reaction mixtures is now an established practice. However, the information that can be obtained is limited to elemental analysis and possibly to fragmentation behavior, which is often challenging to analyze. In order to extend the available experimental information, different types of spectroscopy in the infrared and visible region have been combined with mass spectrometry. Spectroscopy of mass selected ions usually utilizes the powerful sensitivity of mass spectrometers, and the absorption of photons is not detected as such but rather translated to mass changes. One approach to accomplish such spectroscopy involves loosely binding a tag to an ion that will be removed by absorption of one photon. We have constructed an ion trapping instrument capable of reaching temperatures that are sufficiently low to enable tagging by helium atoms in situ, thus permitting infrared photodissociation spectroscopy (IRPD) to be carried out. While tagging by larger rare gas atoms, such as neon or argon is also possible, these may cause significant structural changes to small and reactive species, making the use of helium highly beneficial. We discuss the "innocence" of helium as a tag in ion spectroscopy using several case studies. It is shown that helium tagging is effectively innocent when used with benzene dications, not interfering with their structure or IRPD spectrum. We have also provided a case study where we can see that despite its minimal size there are systems where He has a huge effect. A strong influence of the He tagging was shown in the IRPD spectra of HCCl(2+) where large spectral shifts were observed. While the presented systems are rather small, they involve the formation of mixtures of isomers. We have therefore implemented two-color experiments where one laser is employed to selectively deplete a mixture by one (or more) isomer allowing helium tagging IRPD spectra of the remaining isomer(s) to be recorded via the second laser. Our experimental setup, based on a linear wire quadrupole ion trap, allows us to deplete almost 100% of all helium tagged ions in the trap. Using this special feature, we have developed attenuation experiments for determination of absolute photofragmentation cross sections. At the same time, this approach can be used to estimate the representation of isomers in a mixture. The ultimate aim is the routine use of this instrument and technique to study a wide range of reaction intermediates in catalysis. To this end, we present a study of hypervalent iron(IV)-oxo complexes ([(L)Fe(O)(NO3)](+)). We show that we can spectroscopically differentiate iron complexes with S = 1 and S = 2 according to the stretching vibrations of a nitrate counterion.

A technique for rapid source apportionment applied to ambient organic aerosol measurements from a thermal desorption aerosol gas chromatograph (TAG)

DOE PAGES

Zhang, Yaping; Williams, Brent J.; Goldstein, Allen H.; ...

2016-11-25

Here, we present a rapid method for apportioning the sources of atmospheric organic aerosol composition measured by gas chromatography–mass spectrometry methods. Here, we specifically apply this new analysis method to data acquired on a thermal desorption aerosol gas chromatograph (TAG) system. Gas chromatograms are divided by retention time into evenly spaced bins, within which the mass spectra are summed. A previous chromatogram binning method was introduced for the purpose of chromatogram structure deconvolution (e.g., major compound classes) (Zhang et al., 2014). Here we extend the method development for the specific purpose of determining aerosol samples' sources. Chromatogram bins are arrangedmore » into an input data matrix for positive matrix factorization (PMF), where the sample number is the row dimension and the mass-spectra-resolved eluting time intervals (bins) are the column dimension. Then two-dimensional PMF can effectively do three-dimensional factorization on the three-dimensional TAG mass spectra data. The retention time shift of the chromatogram is corrected by applying the median values of the different peaks' shifts. Bin width affects chemical resolution but does not affect PMF retrieval of the sources' time variations for low-factor solutions. A bin width smaller than the maximum retention shift among all samples requires retention time shift correction. A six-factor PMF comparison among aerosol mass spectrometry (AMS), TAG binning, and conventional TAG compound integration methods shows that the TAG binning method performs similarly to the integration method. However, the new binning method incorporates the entirety of the data set and requires significantly less pre-processing of the data than conventional single compound identification and integration. In addition, while a fraction of the most oxygenated aerosol does not elute through an underivatized TAG analysis, the TAG binning method does have the ability to achieve molecular level resolution on other bulk aerosol components commonly observed by the AMS.« less

RAMTaB: Robust Alignment of Multi-Tag Bioimages

PubMed Central

Raza, Shan-e-Ahmed; Humayun, Ahmad; Abouna, Sylvie; Nattkemper, Tim W.; Epstein, David B. A.; Khan, Michael; Rajpoot, Nasir M.

2012-01-01

Background In recent years, new microscopic imaging techniques have evolved to allow us to visualize several different proteins (or other biomolecules) in a visual field. Analysis of protein co-localization becomes viable because molecules can interact only when they are located close to each other. We present a novel approach to align images in a multi-tag fluorescence image stack. The proposed approach is applicable to multi-tag bioimaging systems which (a) acquire fluorescence images by sequential staining and (b) simultaneously capture a phase contrast image corresponding to each of the fluorescence images. To the best of our knowledge, there is no existing method in the literature, which addresses simultaneous registration of multi-tag bioimages and selection of the reference image in order to maximize the overall overlap between the images. Methodology/Principal Findings We employ a block-based method for registration, which yields a confidence measure to indicate the accuracy of our registration results. We derive a shift metric in order to select the Reference Image with Maximal Overlap (RIMO), in turn minimizing the total amount of non-overlapping signal for a given number of tags. Experimental results show that the Robust Alignment of Multi-Tag Bioimages (RAMTaB) framework is robust to variations in contrast and illumination, yields sub-pixel accuracy, and successfully selects the reference image resulting in maximum overlap. The registration results are also shown to significantly improve any follow-up protein co-localization studies. Conclusions For the discovery of protein complexes and of functional protein networks within a cell, alignment of the tag images in a multi-tag fluorescence image stack is a key pre-processing step. The proposed framework is shown to produce accurate alignment results on both real and synthetic data. Our future work will use the aligned multi-channel fluorescence image data for normal and diseased tissue specimens to analyze molecular co-expression patterns and functional protein networks. PMID:22363510

A technique for rapid source apportionment applied to ambient organic aerosol measurements from a thermal desorption aerosol gas chromatograph (TAG)

NASA Astrophysics Data System (ADS)

Zhang, Yaping; Williams, Brent J.; Goldstein, Allen H.; Docherty, Kenneth S.; Jimenez, Jose L.

2016-11-01

We present a rapid method for apportioning the sources of atmospheric organic aerosol composition measured by gas chromatography-mass spectrometry methods. Here, we specifically apply this new analysis method to data acquired on a thermal desorption aerosol gas chromatograph (TAG) system. Gas chromatograms are divided by retention time into evenly spaced bins, within which the mass spectra are summed. A previous chromatogram binning method was introduced for the purpose of chromatogram structure deconvolution (e.g., major compound classes) (Zhang et al., 2014). Here we extend the method development for the specific purpose of determining aerosol samples' sources. Chromatogram bins are arranged into an input data matrix for positive matrix factorization (PMF), where the sample number is the row dimension and the mass-spectra-resolved eluting time intervals (bins) are the column dimension. Then two-dimensional PMF can effectively do three-dimensional factorization on the three-dimensional TAG mass spectra data. The retention time shift of the chromatogram is corrected by applying the median values of the different peaks' shifts. Bin width affects chemical resolution but does not affect PMF retrieval of the sources' time variations for low-factor solutions. A bin width smaller than the maximum retention shift among all samples requires retention time shift correction. A six-factor PMF comparison among aerosol mass spectrometry (AMS), TAG binning, and conventional TAG compound integration methods shows that the TAG binning method performs similarly to the integration method. However, the new binning method incorporates the entirety of the data set and requires significantly less pre-processing of the data than conventional single compound identification and integration. In addition, while a fraction of the most oxygenated aerosol does not elute through an underivatized TAG analysis, the TAG binning method does have the ability to achieve molecular level resolution on other bulk aerosol components commonly observed by the AMS.

Genome-Wide Analysis of Differentially Expressed Genes Relevant to Rhizome Formation in Lotus Root (Nelumbo nucifera Gaertn)

PubMed Central

Yin, Jingjing; Li, Liangjun; Chen, Xuehao

2013-01-01

Lotus root is a popular wetland vegetable which produces edible rhizome. At the molecular level, the regulation of rhizome formation is very complex, which has not been sufficiently addressed in research. In this study, to identify differentially expressed genes (DEGs) in lotus root, four libraries (L1 library: stolon stage, L2 library: initial swelling stage, L3 library: middle swelling stage, L4: later swelling stage) were constructed from the rhizome development stages. High-throughput tag-sequencing technique was used which is based on Solexa Genome Analyzer Platform. Approximately 5.0 million tags were sequenced, and 4542104, 4474755, 4777919, and 4750348 clean tags including 151282, 137476, 215872, and 166005 distinct tags were obtained after removal of low quality tags from each library respectively. More than 43% distinct tags were unambiguous tags mapping to the reference genes, and 40% were unambiguous tag-mapped genes. From L1, L2, L3, and L4, total 20471, 18785, 23448, and 21778 genes were annotated, after mapping their functions in existing databases. Profiling of gene expression in L1/L2, L2/L3, and L3/L4 libraries were different among most of the selected 20 DEGs. Most of the DEGs in L1/L2 libraries were relevant to fiber development and stress response, while in L2/L3 and L3/L4 libraries, major of the DEGs were involved in metabolism of energy and storage. All up-regulated transcriptional factors in four libraries and 14 important rhizome formation-related genes in four libraries were also identified. In addition, the expression of 9 genes from identified DEGs was performed by qRT-PCR method. In a summary, this study provides a comprehensive understanding of gene expression during the rhizome formation in lotus root. PMID:23840598

Emigration of Natural and Hatchery Chinook Salmon and Steelhead Smolts from the Imnaha River, Oregon, October 20, 1999 to June 15, 2000 : 2000 Annual Report.

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

Cleary, Peter J.

2002-12-01

This report details the smolt performance of natural and hatchery chinook salmon and steelhead from the Imnaha River to the Snake River and Columbia River dams during migration year 2000. Flow conditions in the Imnaha River and Snake River were appreciably lower during May and June in 2000, compared to historic levels at gauging stations, but flow conditions in the Imnaha and Snake River were above average during April. Overall, water conditions for the entire Columbia River were characterized by the Fish Passage Center as below normal levels. Spill occurred continuously at Lower Granite Dam (LGR), Little Goose Dam (LGO),more » and Lower Monumental Dam (LMO) from April 5, April 10, and April 4, respectively, to June 20, and encompassed the periods of migration of Imnaha River juvenile chinook salmon and steelhead, with a few exceptions. Outflow in the tailraces of LGR, LGO, and LMO decreased in May and June while temperatures increased. Chinook salmon and steelhead were captured using rotary screw traps at river kilometer (rkm) 74 and 7 during the fall from October 20 to November 24, 1999, and during the spring period from February 26 to June 15, 2000, at rkm 7. Spring trapping information was reported weekly to the Fish Passage Center's Smolt Monitoring Program. A portion of these fish were tagged weekly with passive integrated transponder (PIT) tags and were detected migrating past interrogation sites at Snake River and Columbia River dams. Survival of PIT tagged fish was estimated with the Survival Using Proportional Hazards model (SURPH model). Estimated survival of fall tagged natural chinook (with {+-} 95% confidence intervals in parenthesis) from the upper Imnaha (rkm 74) to LGR was 29.6% ({+-} 2.8 ). Natural chinook salmon tagged in the fall in the lower Imnaha River at rkm 7, which over wintered in the Snake River, had an estimated survival of 36.8% ({+-} 2.9%) to LGR. Spring tagged natural chinook salmon from the lower site had an estimated survival of 84.8% ({+-} 2.6%) to LGR. The season wide survival of spring tagged natural chinook salmon smolts from release in the Imnaha River to McNary Dam (MCN) was 67.9% ({+-} 6.3%). Post release survival of hatchery chinook salmon smolts, from release at the Imnaha River acclimation facility to the lower Imnaha River trap, was estimated at 94.7% ({+-} 4.7%). Hatchery chinook salmon, PIT tagged and released at the lower Imnaha River trap, had an estimated survival of 75.0% ({+-} 4.2%) to LGR. Estimated survival of hatchery chinook salmon smolts from the Imnaha River to McNary Dam (MCN) was 54.1% ({+-} 9.7%). Natural steelhead smolts had an estimated survival of 84.4% ({+-} 2.7%) to LGR and a survival estimate of 49.9% ({+-}12.2%) from the lower Imnaha River trap to MCN. The estimated survival of hatchery steelhead smolts to LGR was 85.8 ({+-} 2.4) and the survival from release to MCN was 40.2% ({+-}12.5%).« less

Automated tagging of pharmaceutically active thiols under flow conditions using monobromobimane.

PubMed

Tzanavaras, Paraskevas D; Karakosta, Theano D

2011-03-25

The thiol-specific derivatization reagent monobromobimane (MBB) is applied--for the first time--under flow conditions. Sequential injection analysis allows the handling of precise volumes of the reagent in the micro-liter range. The effect of the main chemical and instrumental variables was investigated using captopril (CAP), N-acetylcysteine (NAC) and penicillamine (PEN) as representative pharmaceutically active thiols. Previously reported hydrolysis of MBB due to interaction with nucleophilic components of the buffers was avoided kinetically under flow conditions. The proposed analytical scheme is suitable for the fluorimetric determination of thiols at a sampling rate of 36 h(-1). Copyright © 2010 Elsevier B.V. All rights reserved.

An edge-based solution-adaptive method applied to the AIRPLANE code

NASA Technical Reports Server (NTRS)

Biswas, Rupak; Thomas, Scott D.; Cliff, Susan E.

1995-01-01

Computational methods to solve large-scale realistic problems in fluid flow can be made more efficient and cost effective by using them in conjunction with dynamic mesh adaption procedures that perform simultaneous coarsening and refinement to capture flow features of interest. This work couples the tetrahedral mesh adaption scheme, 3D_TAG, with the AIRPLANE code to solve complete aircraft configuration problems in transonic and supersonic flow regimes. Results indicate that the near-field sonic boom pressure signature of a cone-cylinder is improved, the oblique and normal shocks are better resolved on a transonic wing, and the bow shock ahead of an unstarted inlet is better defined.

The use of tags and tag clouds to discern credible content in online health message forums.

PubMed

O'Grady, Laura; Wathen, C Nadine; Charnaw-Burger, Jill; Betel, Lisa; Shachak, Aviv; Luke, Robert; Hockema, Stephen; Jadad, Alejandro R

2012-01-01

Web sites with health-oriented content are potentially harmful if inaccurate or inappropriate medical information is used to make health-related decisions. Checklists, rating systems and guidelines have been developed to help people determine what is credible, but recent Internet technologies emphasize applications that are collaborative in nature, including tags and tag clouds, where site users 'tag' or label online content, each using their own labelling system. Concepts such as the date, reference, author, testimonial and quotations are considered predictors of credible content. An understanding of these descriptive tools, how they relate to the depiction of credibility and how this relates to overall efforts to label data in relation to the semantic web has yet to emerge. This study investigates how structured (pre-determined) and unstructured (user-generated) tags and tag clouds with a multiple word search feature are used by participants to assess credibility of messages posted in online message forums. The targeted respondents were those using web sites message forums for disease self-management. We also explored the relevancy of our findings to the labelling or indexing of data in the context of the semantic web. Diabetes was chosen as the content area in this study, since (a) this is a condition with increasing prevalence and (b) diabetics have been shown to actively use the Internet to manage their condition. From January to March 2010 participants were recruited using purposive sampling techniques. A screening instrument was used to determine eligibility. The study consisted of a demographic and computer usage survey, a series of usability tests and an interview. We tested participants (N=22) on two scenarios, each involving tasks that assessed their ability to tag content and search using a tag cloud that included six structured credibility terms (statistics, date, reference, author, testimonial and quotations). MORAE Usability software (version 3.1) was employed to record participants' use of the study environment. The surveys were analyzed using SPSS version 17. Interviews with participants were transcribed, coded and analyzed using thematic text analysis with the aid of NVivo8. Most participants had experience with Internet resources. However, less than one quarter of this sample had seen or used tags or a tag clouds. The ways in which participants used tags to label the content posted in the message forums varied. Some participants were tagging the information for their own subsequent use, whereas others viewed this process from the perspective of others: they tagged the content in ways that they thought other users would find beneficial. Many participants did not use the structured credibility tags when asked to search for credible content. The interviews corroborated these findings by confirming participants were not considering credibility foremost when tagging. Many participants in this study focused on assessing whether the information was relevant to their current circumstances, after which they would proceed to determine its credibility by corroborating with other sources. The use of structured tags to label information may not be a useful way to encourage the use of tagging, or to indicate credibility in this context. Current applications used in the semantic web automate this process. Therefore it may be useful to engage consumers of online content, in particular health-related content, to be more directly involved in the annotation of this content. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

Organic and inorganic decomposition products from the thermal desorption of atmospheric particles

NASA Astrophysics Data System (ADS)

Williams, B. J.; Zhang, Y.; Zuo, X.; Martinez, R. E.; Walker, M. J.; Kreisberg, N. M.; Goldstein, A. H.; Docherty, K. S.; Jimenez, J. L.

2015-12-01

Atmospheric aerosol composition is often analyzed using thermal desorption techniques to evaporate samples and deliver organic or inorganic molecules to various designs of detectors for identification and quantification. The organic aerosol (OA) fraction is composed of thousands of individual compounds, some with nitrogen- and sulfur-containing functionality, and often contains oligomeric material, much of which may be susceptible to decomposition upon heating. Here we analyze thermal decomposition products as measured by a thermal desorption aerosol gas chromatograph (TAG) capable of separating thermal decomposition products from thermally stable molecules. The TAG impacts particles onto a collection and thermal desorption (CTD) cell, and upon completion of sample collection, heats and transfers the sample in a helium flow up to 310 °C. Desorbed molecules are refocused at the head of a GC column that is held at 45 °C and any volatile decomposition products pass directly through the column and into an electron impact quadrupole mass spectrometer (MS). Analysis of the sample introduction (thermal decomposition) period reveals contributions of NO+ (m/z 30), NO2+ (m/z 46), SO+ (m/z 48), and SO2+ (m/z 64), derived from either inorganic or organic particle-phase nitrate and sulfate. CO2+ (m/z 44) makes up a major component of the decomposition signal, along with smaller contributions from other organic components that vary with the type of aerosol contributing to the signal (e.g., m/z 53, 82 observed here for isoprene-derived secondary OA). All of these ions are important for ambient aerosol analyzed with the aerosol mass spectrometer (AMS), suggesting similarity of the thermal desorption processes in both instruments. Ambient observations of these decomposition products compared to organic, nitrate, and sulfate mass concentrations measured by an AMS reveal good correlation, with improved correlations for OA when compared to the AMS oxygenated OA (OOA) component. TAG signal found in the traditional compound elution time period reveals higher correlations with AMS hydrocarbon-like OA (HOA) combined with the fraction of OOA that is less oxygenated. Potential to quantify nitrate and sulfate aerosol mass concentrations using the TAG system is explored through analysis of ammonium sulfate and ammonium nitrate standards. While chemical standards display a linear response in the TAG system, re-desorptions of the CTD cell following ambient sample analysis shows some signal carryover on sulfate and organics, and new desorption methods should be developed to improve throughput. Future standards should be composed of complex organic/inorganic mixtures, similar to what is found in the atmosphere, and perhaps will more accurately account for any aerosol mixture effects on compositional quantification.

Organic and inorganic decomposition products from the thermal desorption of atmospheric particles

NASA Astrophysics Data System (ADS)

Williams, Brent J.; Zhang, Yaping; Zuo, Xiaochen; Martinez, Raul E.; Walker, Michael J.; Kreisberg, Nathan M.; Goldstein, Allen H.; Docherty, Kenneth S.; Jimenez, Jose L.

2016-04-01

Atmospheric aerosol composition is often analyzed using thermal desorption techniques to evaporate samples and deliver organic or inorganic molecules to various designs of detectors for identification and quantification. The organic aerosol (OA) fraction is composed of thousands of individual compounds, some with nitrogen- and sulfur-containing functionality and, often contains oligomeric material, much of which may be susceptible to decomposition upon heating. Here we analyze thermal decomposition products as measured by a thermal desorption aerosol gas chromatograph (TAG) capable of separating thermal decomposition products from thermally stable molecules. The TAG impacts particles onto a collection and thermal desorption (CTD) cell, and upon completion of sample collection, heats and transfers the sample in a helium flow up to 310 °C. Desorbed molecules are refocused at the head of a gas chromatography column that is held at 45 °C and any volatile decomposition products pass directly through the column and into an electron impact quadrupole mass spectrometer. Analysis of the sample introduction (thermal decomposition) period reveals contributions of NO+ (m/z 30), NO2+ (m/z 46), SO+ (m/z 48), and SO2+ (m/z 64), derived from either inorganic or organic particle-phase nitrate and sulfate. CO2+ (m/z 44) makes up a major component of the decomposition signal, along with smaller contributions from other organic components that vary with the type of aerosol contributing to the signal (e.g., m/z 53, 82 observed here for isoprene-derived secondary OA). All of these ions are important for ambient aerosol analyzed with the aerosol mass spectrometer (AMS), suggesting similarity of the thermal desorption processes in both instruments. Ambient observations of these decomposition products compared to organic, nitrate, and sulfate mass concentrations measured by an AMS reveal good correlation, with improved correlations for OA when compared to the AMS oxygenated OA (OOA) component. TAG signal found in the traditional compound elution time period reveals higher correlations with AMS hydrocarbon-like OA (HOA) combined with the fraction of OOA that is less oxygenated. Potential to quantify nitrate and sulfate aerosol mass concentrations using the TAG system is explored through analysis of ammonium sulfate and ammonium nitrate standards. While chemical standards display a linear response in the TAG system, redesorptions of the CTD cell following ambient sample analysis show some signal carryover on sulfate and organics, and new desorption methods should be developed to improve throughput. Future standards should be composed of complex organic/inorganic mixtures, similar to what is found in the atmosphere, and perhaps will more accurately account for any aerosol mixture effects on compositional quantification.

Organic and inorganic decomposition products from the thermal desorption of atmospheric particles

DOE PAGES

Williams, Brent J.; Zhang, Yaping; Zuo, Xiaochen; ...

2016-04-11

Here, atmospheric aerosol composition is often analyzed using thermal desorption techniques to evaporate samples and deliver organic or inorganic molecules to various designs of detectors for identification and quantification. The organic aerosol (OA) fraction is composed of thousands of individual compounds, some with nitrogen- and sulfur-containing functionality and, often contains oligomeric material, much of which may be susceptible to decomposition upon heating. Here we analyze thermal decomposition products as measured by a thermal desorption aerosol gas chromatograph (TAG) capable of separating thermal decomposition products from thermally stable molecules. The TAG impacts particles onto a collection and thermal desorption (CTD) cell, and upon completionmore » of sample collection, heats and transfers the sample in a helium flow up to 310 °C. Desorbed molecules are refocused at the head of a gas chromatography column that is held at 45 °C and any volatile decomposition products pass directly through the column and into an electron impact quadrupole mass spectrometer. Analysis of the sample introduction (thermal decomposition) period reveals contributions of NO + ( m/z 30), NO 2 + ( m/z 46), SO + ( m/z 48), and SO 2 + ( m/z 64), derived from either inorganic or organic particle-phase nitrate and sulfate. CO 2 + ( m/z 44) makes up a major component of the decomposition signal, along with smaller contributions from other organic components that vary with the type of aerosol contributing to the signal (e.g., m/z 53, 82 observed here for isoprene-derived secondary OA). All of these ions are important for ambient aerosol analyzed with the aerosol mass spectrometer (AMS), suggesting similarity of the thermal desorption processes in both instruments. Ambient observations of these decomposition products compared to organic, nitrate, and sulfate mass concentrations measured by an AMS reveal good correlation, with improved correlations for OA when compared to the AMS oxygenated OA (OOA) component. TAG signal found in the traditional compound elution time period reveals higher correlations with AMS hydrocarbon-like OA (HOA) combined with the fraction of OOA that is less oxygenated. Potential to quantify nitrate and sulfate aerosol mass concentrations using the TAG system is explored through analysis of ammonium sulfate and ammonium nitrate standards. While chemical standards display a linear response in the TAG system, redesorptions of the CTD cell following ambient sample analysis show some signal carryover on sulfate and organics, and new desorption methods should be developed to improve throughput. Future standards should be composed of complex organic/inorganic mixtures, similar to what is found in the atmosphere, and perhaps will more accurately account for any aerosol mixture effects on compositional quantification.« less

Current radar-responsive tag development activities at Sandia National Laboratories

NASA Astrophysics Data System (ADS)

Ormesher, Richard C.; Plummer, Kenneth W.; Wells, Lars M.

2004-08-01

Over the past ten years, Sandia has developed RF radar responsive tag systems and supporting technologies for various government agencies and industry partners. RF tags can function as RF transmitters or radar transponders that enable tagging, tracking, and location determination functions. Expertise in tag architecture, microwave and radar design, signal analysis and processing techniques, digital design, modeling and simulation, and testing have been directly applicable to these tag programs. In general, the radar responsive tag designs have emphasized low power, small package size, and the ability to be detected by the radar at long ranges. Recently, there has been an interest in using radar responsive tags for Blue Force tracking and Combat ID (CID). The main reason for this interest is to allow airborne surveillance radars to easily distinguish U.S. assets from those of opposing forces. A Blue Force tracking capability would add materially to situational awareness. Combat ID is also an issue, as evidenced by the fact that approximately one-quarter of all U.S. casualties in the Gulf War took the form of ground troops killed by friendly fire. Because the evolution of warfare in the intervening decade has made asymmetric warfare the norm rather than the exception, swarming engagements in which U.S. forces will be freely intermixed with opposing forces is a situation that must be anticipated. Increasing utilization of precision munitions can be expected to drive fires progressively closer to engaged allied troops at times when visual de-confliction is not an option. In view of these trends, it becomes increasingly important that U.S. ground forces have a widely proliferated all-weather radar responsive tag that communicates to all-weather surveillance. The purpose of this paper is to provide an overview of the recent, current, and future radar responsive research and development activities at Sandia National Laboratories that support both the Blue Force Tracking and Combat ID application. Sandia is a multiprogram laboratory operated by Sandia Corporation, a Lockheed Martin Company for the United States Departments of Energy's National Nuclear Security Administration under contract DE-AC04-94AL85000.

Effect of exercise intensity on postprandial lipemia, markers of oxidative stress, and endothelial function after a high-fat meal.

PubMed

Lopes Krüger, Renata; Costa Teixeira, Bruno; Boufleur Farinha, Juliano; Cauduro Oliveira Macedo, Rodrigo; Pinto Boeno, Francesco; Rech, Anderson; Lopez, Pedro; Silveira Pinto, Ronei; Reischak-Oliveira, Alvaro

2016-12-01

The aim of this study was to compare the effects of 2 different exercise intensities on postprandial lipemia, oxidative stress markers, and endothelial function after a high-fat meal (HFM). Eleven young men completed 2-day trials in 3 conditions: rest, moderate-intensity exercise (MI-Exercise) and heavy-intensity exercise (HI-Exercise). Subjects performed an exercise bout or no exercise (Rest) on the evening of day 1. On the morning of day 2, an HFM was provided. Blood was sampled at fasting (0 h) and every hour from 1 to 5 h during the postprandial period for triacylglycerol (TAG), thiobarbituric acid reactive substance (TBARS), and nitrite/nitrate (NOx) concentrations. Flow-mediated dilatation (FMD) was also analyzed. TAG concentrations were reduced in exercise conditions compared with Rest during the postprandial period (P < 0.004). TAG incremental area under the curve (iAUC) was smaller after HI-Exercise compared with Rest (P = 0.012). TBARS concentrations were reduced in MI-Exercise compared with Rest (P < 0.041). FMD was higher in exercise conditions than Rest at 0 h (P < 0.02) and NOx concentrations were enhanced in MI-Exercise compared with Rest at 0 h (P < 0.01). These results suggest that acute exercise can reduce lipemia after an HFM. However, HI-Exercise showed to be more effective in reducing iAUC TAG, which might suggest higher protection against postprandial TAG enhancement. Conversely, MI-Exercise can be beneficial to attenuate the susceptibility of oxidative damage induced by an HFM and to increase endothelial function in the fasted state compared with Rest.

Measurement of the t anti-t production cross-section at √s = 1.96-TeV using lifetime tagging

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

Khanov, Alexander

2004-01-01

A measurement of the tmore » $$\\bar{t}$$ production cross section in the lepton+jets channels with the D0 detector at √s = 1.96 TeV using the lifetime-tagging techniques is presented. The t$$\\bar{t}$$ cross section is estimated from the combination of the e+jets and μ+jets channels. The obtained result σ t$$\\bar{t}$$ = 7.47$$+ 1.22\\atop{-1.14}$$(stat)$$+ 1.65\\atop{-1.03}$$(syst) ± 0.49(lumi) pb is consistent with the Standard Model expectation.« less

Application of coupled affinity-sizing chromatography for the detection of proteolyzed HSA-tagged proteins.

PubMed

London, Anne Serdakowski; Patel, Kunal; Quinn, Lisa; Lemmerer, Martin

2015-04-01

Coupled affinity liquid chromatography and size exclusion chromatography (ALC-SEC) is a technique that has been shown to successfully report product quality of proteins during cell expression and prior to the commencement of downstream processing chromatography steps. This method was applied to monitoring the degradation and subsequent partial remediation of a HSA-tagged protein which showed proteolysis, allowing for rapid cell line development to address this product quality dilemma. This paper outlines the novel application of this method for measuring and addressing protease-induced proteolysis. Copyright © 2014 Elsevier Inc. All rights reserved.

Research and design on system of asset management based on RFID

NASA Astrophysics Data System (ADS)

Guan, Peng; Du, HuaiChang; Jing, Hua; Zhang, MengYue; Zhang, Meng; Xu, GuiXian

2011-10-01

By analyzing the problems in the current assets management, this thesis proposing RFID technology will be applied to asset management in order to improve the management level of automation and information. This paper designed the equipment identification based on 433MHz RFID tag and reader which was deeply studied on the basis of RFID tag and card reader circuits, and this paper also illustrates the system of asset management. The RS232 converts Ethernet is a innovative technology to transfer data to PC monitor software, and implement system of asset management based on WEB techniques (PHP and MySQL).

Use of radio frequency identification (RFID) tags in bedside monitoring of endotracheal tube position.

PubMed

Reicher, Joshua; Reicher, Danielle; Reicher, Murray

2007-06-01

Improper positioning of the endotracheal tube during intubation poses a serious health risk to patients. In one prospective study of 219 critically ill patients, 14% required endotracheal tube repositioning after intubation [Brunel et al. Chest 1989; 96: 1043-1045] While a variety of techniques are used to confirm proper tube placement, a chest X-ray is usually employed for definitive verification. Radio frequency identification (RFID) technology, in which an RFID reader emits and receives a signal from an RFID tag, may be useful in evaluating endotracheal tube position. RFID technology has already been approved for use in humans as a safe and effective tool in a variety of applications. The use of handheld RFID detectors and RFID tag-labeled endotracheal tubes could allow for easy and accurate bedside monitoring of endotracheal tube position, once initial proper placement is confirmed.

Clutter elimination for deep clinical optoacoustic imaging using localised vibration tagging (LOVIT)☆

PubMed Central

Jaeger, Michael; Bamber, Jeffrey C.; Frenz, Martin

2013-01-01

This paper investigates a novel method which allows clutter elimination in deep optoacoustic imaging. Clutter significantly limits imaging depth in clinical optoacoustic imaging, when irradiation optics and ultrasound detector are integrated in a handheld probe for flexible imaging of the human body. Strong optoacoustic transients generated at the irradiation site obscure weak signals from deep inside the tissue, either directly by propagating towards the probe, or via acoustic scattering. In this study we demonstrate that signals of interest can be distinguished from clutter by tagging them at the place of origin with localised tissue vibration induced by the acoustic radiation force in a focused ultrasonic beam. We show phantom results where this technique allowed almost full clutter elimination and thus strongly improved contrast for deep imaging. Localised vibration tagging by means of acoustic radiation force is especially promising for integration into ultrasound systems that already have implemented radiation force elastography. PMID:25302147

Flexible ultra-wideband antenna incorporated with metamaterial structures: multiple notches for chipless RFID application

NASA Astrophysics Data System (ADS)

Jalil, M. E.; Rahim, M. K. A.; Samsuri, N. A.; Dewan, R.; Kamardin, K.

2017-01-01

A coplanar waveguide (CPW) ultra-wideband (UWB) antenna incorporated with metamaterial—split ring resonator structure—that operates from 3.0 to 12.0 GHz is proposed for chipless RFID tag. The 30 mm × 40 mm flexible chipless RFID tag is designed on the fleece substrate ( ɛ r = 1.35, thickness = 1 mm and tan δ = 0.025). A six-slotted modified complementary split ring resonator (MCSRR) is introduced into the ultra-wideband antenna to produce multiple band notches at 3.0, 4.0, 5.0, 6.0 and 7.0 GHz. The frequency shifting technique is introduced for designing a high-capacity chipless RFID tag with compact size. Each MCSRR is able to code in four different allocations (00, 01, 10 and 11). To achieve encoding of 10-bits data (10,234 number), six MCSRRs are proposed with three-slotted MCSRR in the radiator and three-slotted MCSRR in the ground plane.

Glycan reductive isotope labeling for quantitative glycomics.

PubMed

Xia, Baoyun; Feasley, Christa L; Sachdev, Goverdhan P; Smith, David F; Cummings, Richard D

2009-04-15

Many diseases and disorders are characterized by quantitative and/or qualitative changes in complex carbohydrates. Mass spectrometry methods show promise in monitoring and detecting these important biological changes. Here we report a new glycomics method, termed glycan reductive isotope labeling (GRIL), where free glycans are derivatized by reductive amination with the differentially coded stable isotope tags [(12)C(6)]aniline and [(13)C(6)]aniline. These dual-labeled aniline-tagged glycans can be recovered by reverse-phase chromatography and can be quantified based on ultraviolet (UV) absorbance and relative ion abundances. Unlike previously reported isotopically coded reagents for glycans, GRIL does not contain deuterium, which can be chromatographically resolved. Our method shows no chromatographic resolution of differentially labeled glycans. Mixtures of differentially tagged glycans can be directly compared and quantified using mass spectrometric techniques. We demonstrate the use of GRIL to determine relative differences in glycan amount and composition. We analyze free glycans and glycans enzymatically or chemically released from a variety of standard glycoproteins, as well as human and mouse serum glycoproteins, using this method. This technique allows linear relative quantitation of glycans over a 10-fold concentration range and can accurately quantify sub-picomole levels of released glycans, providing a needed advancement in the field of glycomics.

GLYCAN REDUCTIVE ISOTOPE LABELING (GRIL) FOR QUANTITATIVE GLYCOMICS

PubMed Central

Xia, Baoyun; Feasley, Christa L.; Sachdev, Goverdhan P.; Smith, David F.; Cummings, Richard D.

2009-01-01

Many diseases and disorders are characterized by quantitative and/or qualitative changes in complex carbohydrates. Mass spectrometry methods show promise in monitoring and detecting these important biological changes. Here we report a new glycomics method, termed Glycan Reductive Isotope Labeling (GRIL), where free glycans are derivatized by reductive amination with the differentially coded stable isotope tags [12C6]-aniline and [13C6]-aniline. These dual-labeled aniline-tagged glycans can be recovered by reversed-phase chromatography and quantified based on UV-absorbance and relative ion abundances. Unlike previously reported isotopically coded reagents for glycans, GRIL does not contain deuterium, which can be chromatographically resolved. Our method shows no chromatographic resolution of differentially labeled glycans. Mixtures of differentially tagged glycans can be directly compared and quantified using mass spectrometric techniques. We demonstrate the use of GRIL to determine relative differences in glycan amount and composition. We analyze free glycans and glycans enzymatically or chemically released from a variety of standard glycoproteins, as well as human and mouse serum glycoproteins using this method. This technique allows for linear, relative quantitation of glycans over a 10-fold concentration range and can accurately quantify sub-picomole levels of released glycans, providing a needed advancement in the field of Glycomics. PMID:19454239

Easy detection of multiple Alexandrium species using DNA chromatography chip.

PubMed

Nagai, Satoshi; Miyamoto, Shigehiko; Ino, Keita; Tajimi, Seisuke; Nishi, Hiromi; Tomono, Jun

2016-01-01

In this study, the Kaneka DNA chromatography chip (KDCC) for the Alexandrium species was successfully developed for simultaneous detection of five Alexandrium species. This method utilizes a DNA-DNA hybridization technology. In the PCR process, specifically designed tagged-primers are used, i.e. a forward primer consisting of a tag domain, which can conjugate with gold nanocolloids on the chip, and a primer domain, which can anneal/amplify the target sequence. However, the reverse primer consists of a tag domain, which can hybridize to the solid-phased capture probe on the chip, and a primer domain, which can anneal/amplify the target sequence. As a result, a red line that originates from gold nanocolloids appears as a positive signal on the chip, and the amplicon is detected visually by the naked eye. This technique is simple, because it is possible to visually detect the target species soon after (<5min) the application of 2μL of PCR amplicon and 65μL of development buffer to the sample pad of the chip. Further, this technique is relatively inexpensive and does not require expensive laboratory equipment, such as real-time Q-PCR machines or DNA microarray detectors, but a thermal cycler. Regarding the detection limit of KDCC for the five Alexandrium species, it varied among species and it was <0.1-10pg and equivalent to 5-500 copies of rRNA genes, indicating that the technique is sensitive enough for practical use to detect several cells of the target species from 1L of seawater. The detection sensitivity of KDCC was also evaluated with two different techniques, i.e. a multiplex-PCR and a digital DNA hybridization by digital DNA chip analyzer (DDCA), using natural plankton assemblages. There was no significant difference in the detection sensitivity among the three techniques, suggesting KDCC can be readily used to monitor the HAB species. Copyright © 2015 Elsevier B.V. All rights reserved.

Search for DQ2.5 and DQ8 alleles using a lower cost technique in patients with type 1 diabetes and celiac disease in a population of southern Brazil.

PubMed

Bastos, Marília D; Kowalski, Thayne W; Puñales, Márcia; Tschiedel, Balduíno; Mariath, Luiza M; Pires, Ana Luiza G; Faccini, Lavínia S; Silveira, Themis R

2017-12-01

To evaluate the frequency of DQ2.5 and DQ8 alleles using the Tag-single-nucleotide polymorphism (Tag-SNP) technique in individuals with type 1 diabetes mellitus (T1DM) and celiac disease (CD) in southern Brazil. In a prospective design, we performed the search for DQA1*0501 and DQB1*0201 alleles for DQ2.5 and DQB1*0302 for DQ8 through Real-Time Polymerase Chain Reaction (RT-PCR) technique, using TaqMan Genotyping Assays (Applied Biosystems, USA). The diagnosis of CD was established by duodenal biopsy and genotypic determination performed by StepOne Software v2.3. Allelic and genotypic frequencies were compared between groups using Chi-square and Fisher's exact tests and the multiple comparisons using Finner's adjustment. Three hundred and sixty two patients with a median age of 14 years were divided into 3 groups: T1DM without CD (264); T1DM with CD (32) and CD without T1DM (66). In 97% of individuals with T1DM and CD and 76% of individuals with CD without T1DM, respectively, the alleles DQ2.5 and/or DQ8 were identified (p < 0.001). DQ2.5 was more common in individuals with CD (p = 0.004) and DQ8 was more common in individuals with type 1 diabetes (p = 0.008). The evaluation of the alleles for DQ2.5 and DQ8 by Tag-SNP technique showed a high negative predictive value among those with T1DM, similar to that described by the conventional technique. The high frequency of DQ8 alleles in individuals with T1DM did not allow differentiating those at higher risk of developing T1DM.

First experimental demonstration of self-synchronous locking of optical coherence by single-detector electronic-frequency tagging of fiber amplifiers

NASA Astrophysics Data System (ADS)

Shay, T. M.; Benham, Vincent; Baker, J. T.; Ward, Benjamin; Sanchez, Anthony D.; Culpepper, Mark A.; Pilkington, D.; Spring, Justin; Nelson, Douglas J.; Lu, Chunte A.

2006-08-01

A novel high accuracy all electronic technique for phase locking arrays of optical fibers is demonstrated. We report the first demonstration of the only electronic phase locking technique that doesn't require a reference beam. The measured phase error is λ/20. Excellent phase locking has been demonstrated for fiber amplifier arrays.

Identification of cellular MMP substrates using quantitative proteomics: isotope-coded affinity tags (ICAT) and isobaric tags for relative and absolute quantification (iTRAQ).

PubMed

Butler, Georgina S; Dean, Richard A; Morrison, Charlotte J; Overall, Christopher M

2010-01-01

Identification of protease substrates is essential to understand the functional consequences of normal proteolytic processing and dysregulated proteolysis in disease. Quantitative proteomics and mass spectrometry can be used to identify protease substrates in the cellular context. Here we describe the use of two protein labeling techniques, Isotope-Coded Affinity Tags (ICAT and Isobaric Tags for Relative and Absolute Quantification (iTRAQ), which we have used successfully to identify novel matrix metalloproteinase (MMP) substrates in cell culture systems (1-4). ICAT and iTRAQ can label proteins and protease cleavage products of secreted proteins, protein domains shed from the cell membrane or pericellular matrix of protease-transfected cells that have accumulated in conditioned medium, or cell surface proteins in membrane preparations; isotopically distinct labels are used for control cells. Tryptic digestion and tandem mass spectrometry of the generated fragments enable sequencing of differentially labeled but otherwise identical pooled peptides. The isotopic tag, which is unique for each label, identifies the peptides originating from each sample, for instance, protease-transfected or control cells, and comparison of the peak areas enables relative quantification of the peptide in each sample. Thus proteins present in altered amounts between protease-expressing and null cells are implicated as protease substrates and can be further validated as such.

A Novel RFID-Based Sensing Method for Low-Cost Bolt Loosening Monitoring.

PubMed

Wu, Jian; Cui, Xingmei; Xu, Yunpeng

2016-01-28

In coal mines, bolt loosening in the cage guide is affected by the harsh environmental factors and cage hoist vibration, leading to significant threats to work safety. It is crucial, to this effect, to successfully detect the status of multipoint bolts of guide structures. This paper proposes a system to monitor bolt status in harsh environments established based on the RFID technique. A proof-of-concept model was demonstrated consisting of a bolt gearing system, passive UHF RFID tags, a reader, and monitoring software. A tinfoil metal film is fixed on the retaining plate and an RFID tag bonded to a large gear, with the bolt to be detected fixed in the center of a smaller gear. The radio-frequency signal cannot be received by the reader if the tag is completely obscured by the tinfoil, and if the bolt is loose, the tag's antenna is exposed when the gear revolves. A radio-frequency signal that carries corresponding bolt's information is transmitted by the RFID tag to the RFID reader due to coil coupling, identifying loose bolt location and reporting them in the software. Confirmatory test results revealed that the system indeed successfully detects bolt loosening and comparative test results (based on a reed switch multipoint bolt loosening monitor system) provided valuable information regarding the strengths and weaknesses of the proposed system.

MAB_3551c encodes the primary triacylglycerol synthase involved in lipid accumulation in Mycobacterium abscessus.

PubMed

Viljoen, Albertus; Blaise, Mickael; de Chastellier, Chantal; Kremer, Laurent

2016-11-01

Slow growing pathogenic mycobacteria utilize host-derived lipids and accumulate large amounts of triacylglycerol (TAG) in the form of intracytoplasmic lipid inclusions (ILI), serving as a source of carbon and energy during prolonged infection. Mycobacterium abscessus is an emerging and rapidly growing species capable to induce severe and chronic pulmonary infections. However, whether M. abscessus, like Mycobacterium tuberculosis, possesses the machinery to acquire and store host lipids, remains unaddressed. Herein, we aimed at deciphering the contribution of the seven putative M. abscessus TAG synthases (Tgs) in TAG synthesis/accumulation thanks to a combination of genetic and biochemical techniques and a well-defined foamy macrophage (FM) model along with electron microscopy. Targeted gene deletion and functional complementation studies identified the MAB_3551c product, Tgs1, as the major Tgs involved in TAG production. Tgs1 exhibits a preference for long acyl-CoA substrates and site-directed mutagenesis demonstrated that His144 and Gln145 are essential for enzymatic activity. Importantly, in the lipid-rich intracellular context of FM, M. abscessus formed large ILI in a Tgs1-dependent manner. This supports the ability of M. abscessus to assimilate host lipids and the crucial role of Tgs1 in intramycobacterial TAG production, which may represent important mechanisms for long-term storage of a rich energy supply. © 2016 John Wiley & Sons Ltd.

In vivo time-gated diffuse correlation spectroscopy at quasi-null source-detector separation.

PubMed

Pagliazzi, M; Sekar, S Konugolu Venkata; Di Sieno, L; Colombo, L; Durduran, T; Contini, D; Torricelli, A; Pifferi, A; Mora, A Dalla

2018-06-01

We demonstrate time domain diffuse correlation spectroscopy at quasi-null source-detector separation by using a fast time-gated single-photon avalanche diode without the need of time-tagging electronics. This approach allows for increased photon collection, simplified real-time instrumentation, and reduced probe dimensions. Depth discriminating, quasi-null distance measurement of blood flow in a human subject is presented. We envision the miniaturization and integration of matrices of optical sensors of increased spatial resolution and the enhancement of the contrast of local blood flow changes.

First identification of excited states in Ba 117 using the recoil- β -delayed proton tagging technique

DOE PAGES

Ding, B.; Liu, Z.; Seweryniak, D.; ...

2017-02-01

Excited states have been observed for the first time in the neutron-deficient nucleus 117Ba using the recoil-decay tagging technique following the heavy-ion fusion-evaporation reaction 64Zn( 58Ni, 2p3n) 117Ba. Prompt γ rays have been assigned to 117Ba through correlations with β-delayed protons following the decay of A = 117 recoils. Through the analysis of the γ–γ coincidence relationships, a high-spin level scheme consisting of two bands has been established in 117Ba. Based on the systematics of the level spacings in the neighboring barium isotopes, the two bands are proposed to have νh 11/2[532]5/2 – and νd 5/2[413]5/2 + configurations, respectively. Lastly,more » the observed band-crossing properties are interpreted in the framework of cranked shell model.« less

Development of a Photothermal Absorbance Detector for Use with Microfluidic Devices

PubMed Central

Dennis, Patty J.; Ferguson Welch, Erin R.; Alarie, Jean Pierre; Ramsey, J. Michael; Jorgenson, James W.

2010-01-01

The development of a photothermal absorbance detector for use with microfluidic devices is described. Unlike thermooptical techniques that rely on measuring refractive index changes, the solution viscosity is probed by continuously monitoring solution conductivity. Platinum electrodes microfabricated on a quartz substrate and bonded to a substrate containing the microchannels enable contact conductivity measurements. The effects of excitation frequency and voltage, electrode spacing, laser power, and laser modulation (chopping) frequency were evaluated experimentally. In the current configuration a limit of detection of 5 nM for DABSYL-tagged glucosamine was obtained using long injections (to give flat-topped peaks). This corresponds to an absorbance of 4.4 × 10−7 AU. Separation and detection of DABSYL-tagged glycine, proline, and tryptophan is also shown to demonstrate the feasibility of the method. In addition, simulations were used to investigate the applicability of the technique to small volume platforms. PMID:20411923

ManiWordle: providing flexible control over Wordle.

PubMed

Koh, Kyle; Lee, Bongshin; Kim, Bohyoung; Seo, Jinwook

2010-01-01

Among the multifarious tag-clouding techniques, Wordle stands out to the community by providing an aesthetic layout, eliciting the emergence of the participatory culture and usage of tag-clouding in the artistic creations. In this paper, we introduce ManiWordle, a Wordle-based visualization tool that revamps interactions with the layout by supporting custom manipulations. ManiWordle allows people to manipulate typography, color, and composition not only for the layout as a whole, but also for the individual words, enabling them to have better control over the layout result. We first describe our design rationale along with the interaction techniques for tweaking the layout. We then present the results both from the preliminary usability study and from the comparative study between ManiWordle and Wordle. The results suggest that ManiWordle provides higher user satisfaction and an efficient method of creating the desired "art work," harnessing the power behind the ever-increasing popularity of Wordle.

Identification techniques for highly boosted W bosons that decay into hadrons

DOE PAGES

Khachatryan, Vardan

2014-12-02

In searches for new physics in the energy regime of the LHC, it is becoming increasingly important to distinguish single-jet objects that originate from the merging of the decay products of W bosons produced with high transverse momenta from jets initiated by single partons. Algorithms are defined to identify such W jets for different signals of interest, using techniques that are also applicable to other decays of bosons to hadrons that result in a single jet, such as those from highly boosted Z and Higgs bosons. The efficiency for tagging W jets is measured in data collected with the CMSmore » detector at a center-of-mass energy of 8 TeV, corresponding to an integrated luminosity of 19.7 fb -1. The performance of W tagging in data is compared with predictions from several Monte Carlo simulators.« less

An analytical platform for mass spectrometry-based identification and chemical analysis of RNA in ribonucleoprotein complexes.

PubMed

Taoka, Masato; Yamauchi, Yoshio; Nobe, Yuko; Masaki, Shunpei; Nakayama, Hiroshi; Ishikawa, Hideaki; Takahashi, Nobuhiro; Isobe, Toshiaki

2009-11-01

We describe here a mass spectrometry (MS)-based analytical platform of RNA, which combines direct nano-flow reversed-phase liquid chromatography (RPLC) on a spray tip column and a high-resolution LTQ-Orbitrap mass spectrometer. Operating RPLC under a very low flow rate with volatile solvents and MS in the negative mode, we could estimate highly accurate mass values sufficient to predict the nucleotide composition of a approximately 21-nucleotide small interfering RNA, detect post-transcriptional modifications in yeast tRNA, and perform collision-induced dissociation/tandem MS-based structural analysis of nucleolytic fragments of RNA at a sub-femtomole level. Importantly, the method allowed the identification and chemical analysis of small RNAs in ribonucleoprotein (RNP) complex, such as the pre-spliceosomal RNP complex, which was pulled down from cultured cells with a tagged protein cofactor as bait. We have recently developed a unique genome-oriented database search engine, Ariadne, which allows tandem MS-based identification of RNAs in biological samples. Thus, the method presented here has broad potential for automated analysis of RNA; it complements conventional molecular biology-based techniques and is particularly suited for simultaneous analysis of the composition, structure, interaction, and dynamics of RNA and protein components in various cellular RNP complexes.

In vivo Raman flow cytometry for real-time detection of carbon nanotube kinetics in lymph, blood, and tissues

PubMed Central

Biris, Alexandru S.; Galanzha, Ekaterina I.; Li, Zhongrui; Mahmood, Meena; Xu, Yang; Zharov, Vladimir P.

2016-01-01

Nanoparticles are intensively being explored as contrast agents for medical diagnostics and therapies using various optical methods. We present the first demonstration of the use of time-resolved Raman spectroscopy for in vivo real-time detection of circulating carbon nanotubes (CNTs) or cancer cells labeled with CNTs in the lymph, blood, and tissues of live animals with fast spectral acquisition times of down to few milliseconds. After intravenously administering CNTs in the tail vein of the rat, this technique provides the ability to detect the circulation of CNTs in the blood microvessels of the intact rat ear. The capability of Raman spectroscopy is also demonstrated to monitor, identify, and image the CNTs during their transportation by lymphatics in the rat ear and mesentery. The strong and specific Raman scattering properties of CNTs make it possible to detect in vitro and in vivo single cancer cells (HeLa) tagged with CNTs. In vivo Raman flow cytometry opens a new avenue for multiparameter analysis of circulating nanoparticles with strong Raman scattering properties and their pharmokinetics in blood and lymph systems. Moreover, this technology has the potential for molecular detection and identification of circulating tumor cells, and infections labeled with CNTs. PMID:19405719

In vivo Raman flow cytometry for real-time detection of carbon nanotube kinetics in lymph, blood, and tissues

NASA Astrophysics Data System (ADS)

Biris, Alexandru S.; Galanzha, Ekaterina I.; Li, Zhongrui; Mahmood, Meena; Xu, Yang; Zharov, Vladimir P.

2009-03-01

Nanoparticles are intensively being explored as contrast agents for medical diagnostics and therapies using various optical methods. We present the first demonstration of the use of time-resolved Raman spectroscopy for in vivo real-time detection of circulating carbon nanotubes (CNTs) or cancer cells labeled with CNTs in the lymph, blood, and tissues of live animals with fast spectral acquisition times of down to few milliseconds. After intravenously administering CNTs in the tail vein of the rat, this technique provides the ability to detect the circulation of CNTs in the blood microvessels of the intact rat ear. The capability of Raman spectroscopy is also demonstrated to monitor, identify, and image the CNTs during their transportation by lymphatics in the rat ear and mesentery. The strong and specific Raman scattering properties of CNTs make it possible to detect in vitro and in vivo single cancer cells (HeLa) tagged with CNTs. In vivo Raman flow cytometry opens a new avenue for multiparameter analysis of circulating nanoparticles with strong Raman scattering properties and their pharmokinetics in blood and lymph systems. Moreover, this technology has the potential for molecular detection and identification of circulating tumor cells, and infections labeled with CNTs.

Movement and habitat use of green sturgeon Acipenser medirostris in the Rogue River, Oregon, USA

USGS Publications Warehouse

Erickson, D.L.; North, J.A.; Hightower, J.E.; Weber, J.; Lauck, L.

2002-01-01

Green sturgeon (Acipenser medirostris) movement patterns and habitat use within the Rogue River, Oregon were evaluated using radio telemetry. Nineteen specimens ranging from 154 to 225 cm total length were caught by gill netting and tagged with radio transmitters during May-July 2000. One tagged green sturgeon was verified as a female near spawning condition. Individual green sturgeons spent more than 6 months in fresh water and traveled as far as river kilometer (rkm) 39.5. Green sturgeon preferred specific holding sites within the Rogue River during summer and autumn months. These sites were typically deep (> 5 m) low-gradient reaches or off-channel coves. Home ranges within holding sites were restricted. All tagged individuals emigrated from the system to the sea during the autumn and winter, when water temperatures dropped below 10??C and flows increased. This species is extremely vulnerable to habitat alterations and overfishing because it spawns in only a few North American rivers and individuals reside within extremely small areas for extended periods of time.

Magnetic tagging of cell-derived microparticles: new prospects for imaging and manipulation of these mediators of biological information.

PubMed

Vats, Nidhi; Wilhelm, Claire; Rautou, Pierre-Emmanuel; Poirier-Quinot, Marie; Péchoux, Christine; Devue, Cécile; Boulanger, Chantal M; Gazeau, Florence

2010-07-01

Submicron membrane fragments termed microparticles (MPs), which are released by apoptotic or activated cells, are newly considered as vectors of biological information and actors of pathology development. We propose the tagging of MPs with magnetic nanoparticles as a new approach allowing imaging, manipulation and targeting of cell-derived MPs. MPs generated in vitro from human endothelial cells or isolated from atherosclerotic plaques were labeled using citrate-coated 8 nm iron-oxide nanoparticles. MPs were tagged with magnetic nanoparticles on their surface and detected as Annexin-V positive by flow cytometry. Labeled MPs could be mobilized, isolated and manipulated at a distance in a magnetic field gradient. Magnetic mobility of labeled MPs was quantified by micromagnetophoresis. Interactions of labeled MPs with endothelial cells could be triggered and modulated by magnetic guidance. Nanoparticles served as tracers at different scales: at the subcellular level by electron microscopy, at the cellular level by histology and at the macroscopic level by MRI. Magnetic labeling of biogenic MPs opens new prospects for noninvasive monitoring and distal manipulations of these biological effectors.

Ultrasound tagged near infrared spectroscopy does not detect hyperventilation-induced reduction in cerebral blood flow.

PubMed

Lund, Anton; Secher, Niels H; Hirasawa, Ai; Ogoh, Shigehiko; Hashimoto, Takeshi; Schytz, Henrik W; Ashina, Messoud; Sørensen, Henrik

2016-01-01

Continuous non-invasive monitoring of cerebral blood flow (CBF) may be important during anaesthesia and several options are available. We evaluated the CerOx monitor that employs ultrasound tagged near infrared spectroscopy to estimate changes in a CBF index (CFI). Seven healthy males (age 21-26 years) hyperventilated and were administered phenylephrine to increase mean arterial pressure by 20-30 mmHg. Frontal lobe tissue oxygenation (ScO2) and CFI were obtained using the CerOx and mean blood flow velocity in the middle cerebral artery (MCAv mean) was determined by transcranial Doppler. Blood flow in the internal and external carotid artery (ICAf and ECAf) was determined using duplex ultrasonography and forehead skin blood flow (SkBF) and oxygenation (S skin O2) by laser Doppler and white light spectroscopy. During hyperventilation MCAv mean and ICAf decreased by 44% (median; interquartile range 40-49; p = 0.016) and 46% (40-53; p = 0.03), respectively. Conversely, CFI increased by 9% (2-31; p = 0.016), while no significant change was observed in ScO2. SkBF increased by 19% (9-53; p = 0.016) and S skin O2 by 6% (1-7; p = 0.047), although ECAf was unchanged. Administration of phenylephrine was not associated with any changes in MCAv mean, ICAf, ECAf, ScO2, SkBF, S skin O2, or CFI. The CerOx was able to detect a stable CBF during administration of phenylephrine. However, during hyperventilation MCAv mean and ICAf decreased while CFI increased, likely due to an increase in superficial tissue oxygenation. Thus, CFI does not provide an unbiased evaluation of changes in CBF.

Functional polymer sheet patterning using microfluidics.

PubMed

Li, Minggan; Humayun, Mouhita; Kozinski, Janusz A; Hwang, Dae Kun

2014-07-22

Poly(dimethylsiloxane) (PDMS)-based microfluidics provide a novel approach to advanced material synthesis. While PDMS has been successfully used in a wide range of industrial applications, due to the weak mechanical property channels generally possess low aspect ratios (AR) and thus produce microparticles with similarly low ARs. By increasing the channel width to nearly 1 cm, AR to 267, and implementing flow lithography, we were able to establish the slit-channel lithography. Not only does this allow us to synthesize sheet materials bearing multiscale features and tunable chemical anisotropy but it also allows us to fabricate functional layered sheet structures in a one-step, high-throughput fashion. We showcased the technique's potential role in various applications, such as the synthesis of planar material with micro- and nanoscale features, surface morphologies, construction of tubular and 3D layered hydrogel tissue scaffolds, and one-step formation of radio frequency identification (RFID) tags. The method introduced offers a novel route to functional sheet material synthesis and sheet system fabrication.

Associated Particle Tagging (APT) in Magnetic Spectrometers

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

Jordan, David V.; Baciak, James E.; Stave, Sean C.

2012-10-16

Summary In Brief The Associated Particle Tagging (APT) project, a collaboration of Pacific Northwest National Laboratory (PNNL), Idaho National Laboratory (INL) and the Idaho State University (ISU)/Idaho Accelerator Center (IAC), has completed an exploratory study to assess the role of magnetic spectrometers as the linchpin technology in next-generation tagged-neutron and tagged-photon active interrogation (AI). The computational study considered two principle concepts: (1) the application of a solenoidal alpha-particle spectrometer to a next-generation, large-emittance neutron generator for use in the associated particle imaging technique, and (2) the application of tagged photon beams to the detection of fissile material via active interrogation.more » In both cases, a magnetic spectrometer momentum-analyzes charged particles (in the neutron case, alpha particles accompanying neutron generation in the D-T reaction; in the tagged photon case, post-bremsstrahlung electrons) to define kinematic properties of the relevant neutral interrogation probe particle (i.e. neutron or photon). The main conclusions of the study can be briefly summarized as follows: Neutron generator: • For the solenoidal spectrometer concept, magnetic field strengths of order 1 Tesla or greater are required to keep the transverse size of the spectrometer smaller than 1 meter. The notional magnetic spectrometer design evaluated in this feasibility study uses a 5-T magnetic field and a borehole radius of 18 cm. • The design shows a potential for 4.5 Sr tagged neutron solid angle, a factor of 4.5 larger than achievable with current API neutron-generator designs. • The potential angular resolution for such a tagged neutron beam can be less than 0.5o for modest Si-detector position resolution (3 mm). Further improvement in angular resolution can be made by using Si-detectors with better position resolution. • The report documents several features of a notional generator design incorporating the alpha-particle spectrometer concept, and outlines challenges involved in the magnetic field design. Tagged photon interrogation: • We investigated a method for discriminating fissile from benign cargo-material response to an energy-tagged photon beam. The method relies upon coincident detection of the tagged photon and a photoneutron or photofission neutron produced in the target material. The method exploits differences in the shape of the neutron production cross section as a function of incident photon energy in order to discriminate photofission yield from photoneutrons emitted by non-fissile materials. Computational tests of the interrogation method as applied to material composition assay of a simple, multi-layer target suggest that the tagged-photon information facilitates precise (order 1% thickness uncertainty) reconstruction of the constituent thicknesses of fissile (uranium) and high-Z (Pb) constituents of the test targets in a few minutes of photon-beam exposure. We assumed an 18-MeV endpoint tagged photon beam for these simulations. • The report addresses several candidate design and data analysis issues for beamline infrastructure required to produce a tagged photon beam in a notional AI-dedicated facility, including the accelerator and tagging spectrometer.« less

Novel techniques and insights into the deployment of pop-up satellite archival tags on a small-bodied deep-water chondrichthyan

NASA Astrophysics Data System (ADS)

Shipley, Oliver N.; Howey, Lucy A.; Tolentino, Emily R.; Jordan, Lance K. B.; Brooks, Edward J.

2017-01-01

Acquiring movement data for small-bodied, deep-water chondrichthyans is challenged by extreme effects of capture and handling stress, and post-release predation, however, it is urgently required to examine important fisheries interactions and assess the ecological role of these species within deep-water food webs. Here we suggest a novel release-cage mechanism to deploy pop-up satellite archival tags, as well as present vertical habitat data for a data-deficient, small-bodied, deep-water bycatch species, the Cuban dogfish (Squalus cubensis). Data were gathered from seven of eight High Rate X-Tags deployed on mature Cuban dogfish in the Exuma Sound, The Bahamas. Recovery periods appeared variable between individuals and are likely driven by capture-and-handling stress and tag burden. Application of the cross-correlation function to time-series depth and temperature data indicated three of the seven individuals suffered mortality through predation, which occurred during daytime, and suggests Cuban dogfish may constitute a proportion of deep-water apex predator diet in the Exuma Sound. Two animals were successfully released via a novel release-cage mechanism and displayed either no, or rapid (<15 mins) vertically stationary recovery periods and were not consumed by predators; data for these individuals were recorded for the entire deployment duration (14 days). Vertical habitat data suggests Cuban dogfish are diel-vertical migrators, similar to other deep-water taxa, and exhibit a relatively broad temperature and depth range, which may be driven by preference for specific bathymetric structures. These techniques provide an important first step into acquiring and presenting vertical habitat data for small-bodied, deep-water chondrichthyans, which can be directly applied to fisheries and ecosystem-based management approaches.

Evaluation of coded wire tags for marking lake trout

USGS Publications Warehouse

Elrod, Joseph H.; Schneider, Clifford P.

1986-01-01

Among hatchery-reared lake trout (Salvelinus namaycush) of the 1979-1982 year classes stocked in New York waters of Lake Ontario, more than 3 million fish were marked with a coded wire tag (CWT) plus an adipose fin clip, and 1.5 million with only conventional fin clips. Altogether, 7,640 tags were recovered from fish collected with bottom trawls and gill nets or caught by anglers during 1980-1983. One person was able to extract and decipher 200 or more CWTs per day with about a 1% error rate in reading and recording codes. Presence of the CWT did not affect growth. The adipose fin clip did not regenerate. The occurrence of fish with an adipose fin clip but no CWT resulted primarily from the regeneration of paired fins among fish marked with a combination of the adipose fin and a paired fin. Loss of CWTs between marking and stocking (generally 4-5 months for fish stocked in spring and 1-8 d for fish stocked in fall) declined from nearly 11% for the 1979 year class stocked as fall fingerlings to less than 3% for the 1981 and 1982 year classes - a difference that primarily reflected improvements in instrumentation and tagging technique. The rate of CWT loss after the marked fish were stocked was probably less than 1% per year. The CWT is a reliable method for marking hatchery-reared lake trout. A large number of experimental groups can be uniquely marked, and fish from each group can be accurately identified throughout their life. Use of this technique should greatly facilitate evaluations of genetic strain, hatchery experience, condition at time of stocking, season of stocking, size at stocking, method of stocking and other factors that affect poststocking survival and performance of lake trout stocked in the Great Lakes.

MilQuant: a free, generic software tool for isobaric tagging-based quantitation.

PubMed

Zou, Xiao; Zhao, Minzhi; Shen, Hongyan; Zhao, Xuyang; Tong, Yuanpeng; Wang, Qingsong; Wei, Shicheng; Ji, Jianguo

2012-09-18

Isobaric tagging techniques such as iTRAQ and TMT are widely used in quantitative proteomics and especially useful for samples that demand in vitro labeling. Due to diversity in choices of MS acquisition approaches, identification algorithms, and relative abundance deduction strategies, researchers are faced with a plethora of possibilities when it comes to data analysis. However, the lack of generic and flexible software tool often makes it cumbersome for researchers to perform the analysis entirely as desired. In this paper, we present MilQuant, mzXML-based isobaric labeling quantitator, a pipeline of freely available programs that supports native acquisition files produced by all mass spectrometer types and collection approaches currently used in isobaric tagging based MS data collection. Moreover, aside from effective normalization and abundance ratio deduction algorithms, MilQuant exports various intermediate results along each step of the pipeline, making it easy for researchers to customize the analysis. The functionality of MilQuant was demonstrated by four distinct datasets from different laboratories. The compatibility and extendibility of MilQuant makes it a generic and flexible tool that can serve as a full solution to data analysis of isobaric tagging-based quantitation. Copyright © 2012 Elsevier B.V. All rights reserved.

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

Kim, Hyo-Seob; Ames Lab., Ames, IA; Dharmaiah, Peyala

(GeTe) x(AgSbTe 2) 100$-$x: TAGS thermoelectrics are an attractive class of materials due to their combination of non-toxicity and good conversion efficiency at mid-temperature ranges. Here in the present work, we have utilized energy and time efficient high-pressure gas atomization and spark-plasma sintering techniques for large-scale preparation of samples with varying composition (i.e., (GeTe) x(AgSbTe 2) 100$-$x where x = 75, 80, 85, and 90). High-temperature x-ray diffraction was used to understand the phase transformation mechanism of the as-atomized powders. Detailed high-resolution transmission electron microscopy of the sintered samples revealed the presence of nanoscale precipitates, antiphase, and twin boundaries. Themore » nanoscale twins and antiphase boundaries serve as phonon scattering centers, leading to the reduction of total thermal conductivity in TAGS-80 and 90 samples. The maximum ZT obtained was 1.56 at 623 K for TAGS-90, which was ~94% improvement compared to values previously reported. The presence of the twin boundaries also resulted in a high fracture toughness (K IC) of the TAGS-90 sample due to inhibition of dislocation movement at the twin boundary.« less

The self-assembly of redox active peptides: Synthesis and electrochemical capacitive behavior.

PubMed

Piccoli, Julia P; Santos, Adriano; Santos-Filho, Norival A; Lorenzón, Esteban N; Cilli, Eduardo M; Bueno, Paulo R

2016-05-01

The present work reports on the synthesis of a redox-tagged peptide with self-assembling capability aiming applications in electrochemically active capacitive surfaces (associated with the presence of the redox centers) generally useful in electroanalytical applications. Peptide containing ferrocene (fc) molecular (redox) group (Ac-Cys-Ile-Ile-Lys(fc)-Ile-Ile-COOH) was thus synthesized by solid phase peptide synthesis (SPPS). To obtain the electrochemically active capacitive interface, the side chain of the cysteine was covalently bound to the gold electrode (sulfur group) and the side chain of Lys was used to attach the ferrocene in the peptide chain. After obtaining the purified redox-tagged peptide, the self-assembly and redox capability was characterized by cyclic voltammetry (CV) and electrochemical impedance-based capacitance spectroscopy techniques. The obtained results confirmed that the redox-tagged peptide was successfully attached by forming an electroactive self-assembled monolayer onto gold electrode. The design of redox active self-assembly ferrocene-tagged peptide is predictably useful in the development of biosensor devices precisely to detect, in a label-free platform, those biomarkers of clinical relevance. © 2016 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 106: 357-367, 2016. © 2016 Wiley Periodicals, Inc.

Triacylglycerol stereospecific analysis and linear discriminant analysis for milk speciation.

PubMed

Blasi, Francesca; Lombardi, Germana; Damiani, Pietro; Simonetti, Maria Stella; Giua, Laura; Cossignani, Lina

2013-05-01

Product authenticity is an important topic in dairy sector. Dairy products sold for public consumption must be accurately labelled in accordance with the contained milk species. Linear discriminant analysis (LDA), a common chemometric procedure, has been applied to fatty acid% composition to classify pure milk samples (cow, ewe, buffalo, donkey, goat). All original grouped cases were correctly classified, while 90% of cross-validated grouped cases were correctly classified. Another objective of this research was the characterisation of cow-ewe milk mixtures in order to reveal a common fraud in dairy field, that is the addition of cow to ewe milk. Stereospecific analysis of triacylglycerols (TAG), a method based on chemical-enzymatic procedures coupled with chromatographic techniques, has been carried out to detect fraudulent milk additions, in particular 1, 3, 5% cow milk added to ewe milk. When only TAG composition data were used for the elaboration, 75% of original grouped cases were correctly classified, while totally correct classified samples were obtained when both total and intrapositional TAG data were used. Also the results of cross validation were better when TAG stereospecific analysis data were considered as LDA variables. In particular, 100% of cross-validated grouped cases were obtained when 5% cow milk mixtures were considered.

A robust and accurate center-frequency estimation (RACE) algorithm for improving motion estimation performance of SinMod on tagged cardiac MR images without known tagging parameters.

PubMed

Liu, Hong; Wang, Jie; Xu, Xiangyang; Song, Enmin; Wang, Qian; Jin, Renchao; Hung, Chih-Cheng; Fei, Baowei

2014-11-01

A robust and accurate center-frequency (CF) estimation (RACE) algorithm for improving the performance of the local sine-wave modeling (SinMod) method, which is a good motion estimation method for tagged cardiac magnetic resonance (MR) images, is proposed in this study. The RACE algorithm can automatically, effectively and efficiently produce a very appropriate CF estimate for the SinMod method, under the circumstance that the specified tagging parameters are unknown, on account of the following two key techniques: (1) the well-known mean-shift algorithm, which can provide accurate and rapid CF estimation; and (2) an original two-direction-combination strategy, which can further enhance the accuracy and robustness of CF estimation. Some other available CF estimation algorithms are brought out for comparison. Several validation approaches that can work on the real data without ground truths are specially designed. Experimental results on human body in vivo cardiac data demonstrate the significance of accurate CF estimation for SinMod, and validate the effectiveness of RACE in facilitating the motion estimation performance of SinMod. Copyright © 2014 Elsevier Inc. All rights reserved.

Ultra-High-Throughput Screening of an In Vitro-Synthesized Horseradish Peroxidase Displayed on Microbeads Using Cell Sorter

PubMed Central

Zhu, Bo; Mizoguchi, Takuro; Kojima, Takaaki; Nakano, Hideo

2015-01-01

The C1a isoenzyme of horseradish peroxidase (HRP) is an industrially important heme-containing enzyme that utilizes hydrogen peroxide to oxidize a wide variety of inorganic and organic compounds for practical applications, including synthesis of fine chemicals, medical diagnostics, and bioremediation. To develop a ultra-high-throughput screening system for HRP, we successfully produced active HRP in an Escherichia coli cell-free protein synthesis system, by adding disulfide bond isomerase DsbC and optimizing the concentrations of hemin and calcium ions and the temperature. The biosynthesized HRP was fused with a single-chain Cro (scCro) DNA-binding tag at its N-terminal and C-terminal sites. The addition of the scCro-tag at both ends increased the solubility of the protein. Next, HRP and its fusion proteins were successfully synthesized in a water droplet emulsion by using hexadecane as the oil phase and SunSoft No. 818SK as the surfactant. HRP fusion proteins were displayed on microbeads attached with double-stranded DNA (containing the scCro binding sequence) via scCro-DNA interactions. The activities of the immobilized HRP fusion proteins were detected with a tyramide-based fluorogenic assay using flow cytometry. Moreover, a model microbead library containing wild type hrp (WT) and inactive mutant (MUT) genes was screened using fluorescence-activated cell-sorting, thus efficiently enriching the WT gene from the 1:100 (WT:MUT) library. The technique described here could serve as a novel platform for the ultra-high-throughput discovery of more useful HRP mutants and other heme-containing peroxidases. PMID:25993095

Prevention of water-contamination of ethanol-saturated dentin and hydrophobic hybrid layers

PubMed Central

Sauro, Salvatore; Watson, Timothy F; Mannocci, Francesco; Tay, Franklin R; Pashley, David H

2013-01-01

SUMMARY Purpose This in vitro study evaluated the amount and the distribution of outward fluid flow that occurred when an experimental etch-and-rinse hydrophobic adhesive was applied to ethanol-saturated dentin before and after oxalate pretreatment. Materials and methods Measurements of dentin permeability were performed under a constant pulpal pressure of 20 cm H2O in deep and middle dentin. A lucifer yellow solution was placed in the pulp chamber to determine the distribution of the water contamination of the hybrid layers. Results The distribution of fluorescence in dentin specimens that were not pretreated with oxalate revealed that the dye permeated around the resin tags and filled the hybrid layer. Dentin specimens pretreated with oxalate prior to resin bonding, showed 80–83% less (p<0.05) water contamination compared to controls. The dentin permeability results obtained before and after oxalate pretreatment showed that oxalate decreased dentin permeability by 98% (p<0.05) compared to acid-etched controls. This prevented outward fluid movement during bonding resulting in better resin sealing of dentin due to the formation of a double seal of resin tags over calcium oxalate crystals in the tubules. Conclusion Outward dentinal fluid flow may contaminate hybrid layers during adhesive bonding procedures. Pretreatment of acid-etched dentin with 3% oxalic acid prior to bonding procedures can prevent outward fluid flow during bonding and water contamination of the hydrophobic hybrid layers. PMID:19701507

Investigation of the feasibility of optical diagnostic measurements at the exit of the SSME

NASA Technical Reports Server (NTRS)

Shirley, John A.; Boedeker, Laurence R.

1993-01-01

Under Contract NAS8-36861 sponsored by NASA Marshall Space Flight Center, the United Technologies Research Center is conducting an investigation of the feasibility of remote optical diagnostics to measure temperature, species concentration and velocity at the exit of the Space Shuttle Main Engine (SSME). This is a two phase study consisting of a conceptual design phase followed by a laboratory experimental investigation. The first task of the conceptual design studies is to screen and evaluate the techniques which can be used for the measurements. The second task is to select the most promising technique or techniques, if as expected, more than one type of measurement must be used to measure all the flow variables of interest. The third task is to examine in detail analytically the capabilities and limitations of the selected technique(s). The results of this study are described in the section of this report entitled Conceptual Design Investigations. The conceptual design studies identified spontaneous Raman scattering and photodissociative flow-tagging for measurements respectively of gas temperature and major species concentration and for velocity. These techniques and others that were considered are described in the section describing the conceptual design. The objective of the second phase of investigations was to investigate experimentally the techniques identified in the first phase. The first task of the experimental feasibility study is to design and assemble laboratory scale experimental apparatus to evaluate the best approaches for SSME exit optical diagnostics for temperature, species concentrations and velocity, as selected in the Phase I conceptual design study. The second task is to evaluate performance, investigate limitations, and establish actual diagnostic capabilities, accuracies and precision for the selected optical systems. The third task is to evaluate design requirements and system trade-offs of conceptual instruments. Spontaneous Raman scattering excited by a KrF excimer laser pulse was investigated for SSME exit plane temperature and major species concentration measurements. The relative concentrations of molecular hydrogen and water vapor would be determined by measuring the integrated Q-branch scattering signals through narrow bandpass filters in front of photomultipliers. The temperature would be determined by comparing the signal from a single hydrogen rotational Raman line to the total hydrogen Q-branch signal. The rotational Raman line would be isolated by a monochromator and detected with a PMT.

Automating Network Node Behavior Characterization by Mining Communication Patterns

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

Carroll, Thomas E.; Chikkagoudar, Satish; Arthur-Durett, Kristine M.

Enterprise networks of scale are complex, dynamic computing environments that respond to evolv- ing business objectives and requirements. Characteriz- ing system behaviors in these environments is essential for network management and cyber security operations. Characterization of system’s communication is typical and is supported using network flow information (NetFlow). Related work has characterized behavior using theoretical graph metrics; results are often difficult to interpret by enterprise staff. We propose a different approach, where flow information is mapped to sets of tags that contextualize the data in terms of network principals and enterprise concepts. Frequent patterns are then extracted and are expressedmore » as behaviors. Behaviors can be com- pared, identifying systems expressing similar behaviors. We evaluate the approach using flow information collected by a third party.« less

X-Ray Fluorescence Solvent Detection at the Substrate-Adhesive Interface

NASA Technical Reports Server (NTRS)

Wurth, Laura; Evans, Kurt; Weber, Bart; Headrick, Sarah

2005-01-01

With environmental regulations limiting the use of volatile organic compounds, low-vapor pressure solvents have replaced traditional degreasing solvents for bond substrate preparation. When used to clean and prepare porous bond substrates such as phenolic composites, low vapor pressure solvents can penetrate deep into substrate pore networks and remain there for extended periods. Trapped solvents can interact with applied adhesives either prior to or during cure, potentially compromising bond properties. Currently, methods for characterizing solvent time-depth profiles in bond substrates are limited to bulk gravimetric or sectioning techniques. While sectioning techniques such as microtome allow construction of solvent depth profiles, their depth resolution and reliability are limited by substrate type. Sectioning techniques are particularly limited near the adhesive-substrate interface where depth resolution is further limited by adhesive-substrate hardness and, in the case of a partially cured adhesive, mechanical properties differences. Additionally, sectioning techniques cannot provide information about lateral solvent diffusion. Cross-section component mapping is an alternative method for measuring solvent migration in porous substrates that eliminates the issues associated with sectioning techniques. With cross-section mapping, the solvent-wiped substrate is sectioned perpendicular rather than parallel to the wiped surface, and the sectioned surface is analyzed for the solvent or solvent components of interest using a two-dimensional mapping or imaging technique. Solvent mapping can be performed using either direct or indirect methods. With a direct method, one or more solvent components are mapped using red or Raman spectroscopy together with a moveable sample stage and/or focal plane array detector. With an indirect method, an elemental "tag" not present in the substrate is added to the solvent before the substrate is wiped. Following cross sectioning, the tag element can then be mapped by its characteristic x-ray emission using either x-ray fluorescence, or electron-beam energy-and wavelength-dispersive x-ray spectrometry. The direct mapping techniques avoid issues of different diffusion or migration rates of solvents and elemental tags, while the indirect techniques avoid spectral resolution issues in cases where solvents and substrates have adjacent or overlapping peaks. In this study, cross-section component indirect mapping is being evaluated as a method for measuring migration of d-limonene based solvents in glass-cloth phenolic composite (GCP) prior to and during subsequent bonding and epoxy adhesive cure.

Large wood budget and transport dynamics on a large river using radio telemetry

USGS Publications Warehouse

Schenk, Edward R.; Moulin, Bertrand; Hupp, Cliff R.; Richte, Jean M.

2014-01-01

Despite the abundance of large wood (LW) river studies there is still a lack of understanding of LW transport dynamics on large low gradient rivers. This study used 290 radio frequency identification tagged (RFID) LW and 54 metal (aluminum) tagged LW, to quantify the percent of in-channel LW that moves per year and what variables play a role in LW transport dynamics. Aluminum tags were installed and monitored on LW in-transit during the rising limb of a flood, the mean distance traveled by those pieces during the week was 13.3 river kilometers (km) with a maximum distance of 72 km. RFID tagged LW moved a mean of 11.9 km/yr with a maximum observed at 101.1 km/yr. Approximately 41% of LW low on the bank moves per year. The high rate of transport and distance traveled is likely due to the lack of interaction between LW floating in the channel and the channel boundaries, caused primarily by the width of the channel relative to length of the LW. Approximately 80% of the RFID tags moved past a fixed reader during the highest 20% of river stage per year. LW transport and logjam dynamics are complicated at high flows as pieces form temporary jams that continually expand and contract. Unlike most other studies, key members that create a logjam were defined more by stability than jam size or channel/hydrologic conditions. Finally, using an existing geomorphic database for the river, and data from this study, we were able to develop a comprehensive LW budget showing that 5% of the in-channel LW population turns over each year (input from mass wasting and fluvial erosion equals burial, decomposition, and export out of system) and another 16% of the population moving within the system.

Characterization of SV-40 Tag rats as a model to study prostate cancer

PubMed Central

2009-01-01

Background Prostate cancer is the second most frequently diagnosed cancer in men. Animal models that closely mimic clinical disease in humans are invaluable tools in the fight against prostate cancer. Recently, a Simian Virus-40 T-antigen (SV-40 Tag) targeted probasin promoter rat model was developed. This model, however, has not been extensively characterized; hence we have investigated the ontogeny of prostate cancer and determined the role of sex steroid receptor and insulin-like growth factor-1 (IGF-1) signaling proteins in the novel SV-40 Tag rat. Methods The SV-40 Tag rat was histopathologically characterized for time to tumor development, incidence and multiplicity and in the ventral, dorsal, lateral and anterior lobes of the prostate. Immunoassay techniques were employed to measure cell proliferation, apoptosis, and sex steroid receptor and growth factor signaling-related proteins. Steroid hormone concentrations were measured via coated well enzyme linked immunosorbent assay (ELISA) kits. Results Prostatic intraepithelial neoplasia (PIN) and well-differentiated prostate cancer developed as early as 2 and 10 weeks of age, respectively in the ventral prostate (VP) followed by in the dorsolateral (DLP). At 8 weeks of age, testosterone and dihydrotestosterone (DHT) concentrations in SV-40 Tag rats were increased when compared to non-transgenic rats. High cell proliferation and apoptotic indices were found in VP and DLP of transgenic rats. Furthermore, we observed increased protein expression of androgen receptor, IGF-1, IGF-1 receptor, and extracellular signal-regulated kinases in the prostates of SV-40 Tag rats. Conclusion The rapid development of PIN and prostate cancer in conjunction with the large prostate size makes the SV-40 Tag rat a useful model for studying prostate cancer. This study provides evidence of the role of sex steroid and growth factor proteins in prostate cancer development and defines appropriate windows of opportunity for preclinical trials and aids in the rational design of chemoprevention, intervention, regression, and therapeutic studies using prostate cancer rodent models. PMID:19171036

Transcriptome Analysis of Gene Expression during Chinese Water Chestnut Storage Organ Formation

PubMed Central

Chen, Sainan; Wang, Yan; Yu, Meizhen; Chen, Xuehao; Li, Liangjun; Yin, Jingjing

2016-01-01

The product organ (storage organ; corm) of the Chinese water chestnut has become a very popular food in Asian countries because of its unique nutritional value. Corm formation is a complex biological process, and extensive whole genome analysis of transcripts during corm development has not been carried out. In this study, four corm libraries at different developmental stages were constructed, and gene expression was identified using a high-throughput tag sequencing technique. Approximately 4.9 million tags were sequenced, and 4,371,386, 4,372,602, 4,782,494, and 5,276,540 clean tags, including 119,676, 110,701, 100,089, and 101,239 distinct tags, respectively, were obtained after removal of low-quality tags from each library. More than 39% of the distinct tags were unambiguous and could be mapped to reference genes, while 40% were unambiguous tag-mapped genes. After mapping their functions in existing databases, a total of 11,592, 10,949, 10,585, and 7,111 genes were annotated from the B1, B2, B3, and B4 libraries, respectively. Analysis of the differentially expressed genes (DEGs) in B1/B2, B2/B3, and B3/B4 libraries showed that most of the DEGs at the B1/B2 stages were involved in carbohydrate and hormone metabolism, while the majority of DEGs were involved in energy metabolism and carbohydrate metabolism at the B2/B3 and B3/B4 stages. All of the upregulated transcription factors and 9 important genes related to product organ formation in the above four stages were also identified. The expression changes of nine of the identified DEGs were validated using a quantitative PCR approach. This study provides a comprehensive understanding of gene expression during corm formation in the Chinese water chestnut. PMID:27716802

Development and in silico evaluation of large-scale metabolite identification methods using functional group detection for metabolomics

PubMed Central

Mitchell, Joshua M.; Fan, Teresa W.-M.; Lane, Andrew N.; Moseley, Hunter N. B.

2014-01-01

Large-scale identification of metabolites is key to elucidating and modeling metabolism at the systems level. Advances in metabolomics technologies, particularly ultra-high resolution mass spectrometry (MS) enable comprehensive and rapid analysis of metabolites. However, a significant barrier to meaningful data interpretation is the identification of a wide range of metabolites including unknowns and the determination of their role(s) in various metabolic networks. Chemoselective (CS) probes to tag metabolite functional groups combined with high mass accuracy provide additional structural constraints for metabolite identification and quantification. We have developed a novel algorithm, Chemically Aware Substructure Search (CASS) that efficiently detects functional groups within existing metabolite databases, allowing for combined molecular formula and functional group (from CS tagging) queries to aid in metabolite identification without a priori knowledge. Analysis of the isomeric compounds in both Human Metabolome Database (HMDB) and KEGG Ligand demonstrated a high percentage of isomeric molecular formulae (43 and 28%, respectively), indicating the necessity for techniques such as CS-tagging. Furthermore, these two databases have only moderate overlap in molecular formulae. Thus, it is prudent to use multiple databases in metabolite assignment, since each major metabolite database represents different portions of metabolism within the biosphere. In silico analysis of various CS-tagging strategies under different conditions for adduct formation demonstrate that combined FT-MS derived molecular formulae and CS-tagging can uniquely identify up to 71% of KEGG and 37% of the combined KEGG/HMDB database vs. 41 and 17%, respectively without adduct formation. This difference between database isomer disambiguation highlights the strength of CS-tagging for non-lipid metabolite identification. However, unique identification of complex lipids still needs additional information. PMID:25120557

In-Depth Lipidomic Analysis of Molecular Species of Triacylglycerides, Diacylglycerides, Glycerophospholipids, and Sphingolipids of Buttermilk by GC-MS/FID, HPLC-ELSD, and UPLC-QToF-MS

PubMed Central

Castro-Gómez, Pilar; Montero, Olimpio; Fontecha, Javier

2017-01-01

Buttermilk, a byproduct of butter manufacturing, has gained considerable attention due to its high concentration of polar lipids as phospho- and sphingolipids from the milk fat globule membrane (MFGM). These polar lipids (PLs) are essential components of all cellular membranes and exert a variety of indispensable metabolic, neurological, and intracellular signaling processes. Despite its importance, there are few research studies that report a comprehensive characterization of the lipid molecular species of MFGM that could contribute to a better understanding of their putative healthful activities. In this study, procedures such as pressurized liquid extraction of polar and nonpolar lipids and their fractionation by flash chromatography have been carried out. The obtained fractions were submitted to an exhaustive characterization from a lipidomic point of view. The characterization includes new data about the identification and quantification of triacylglycerides (TAG), diacylglycerides (DAG), and phospho- and sphingolipids using different chromatographic techniques. The fatty acid profile was comparable to that of the milk fat but with a highly diverse composition of fatty acids. Molecular species have also been determined by using ultra-high performance liquid chromatography/quadruple-time-of-flight mass spectrometry (UPLC/QToF-MS). The TAG (16:0/16:0/6:0) and TAG (16:0/16:0/8:0) were the predominant saturated TAG species and TAG (14:0/18:1/16:0) and TAG (16:0/16:0/18:1) presented the highest content of monounsaturated TAG species. Furthermore; over 30 molecular species of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylinositol (PI) could be identified within PL, with PC (16:0/18:1) being the most abundant species. Whereas C16:0 was found to be the preferred FA in TAGs, it was C18:1 in PLs. Several ganglioside species have also been characterized with d18:1 ceramide moiety and secondary acyl chains ranging from C20:0 to C26:1. This approach could broaden the applications of high-resolution mass spectrometry for a better understanding of the role of MFGM and its functionality. PMID:28287421

Live-cell imaging of migrating cells expressing fluorescently-tagged proteins in a three-dimensional matrix.

PubMed

Shih, Wenting; Yamada, Soichiro

2011-12-22

Traditionally, cell migration has been studied on two-dimensional, stiff plastic surfaces. However, during important biological processes such as wound healing, tissue regeneration, and cancer metastasis, cells must navigate through complex, three-dimensional extracellular tissue. To better understand the mechanisms behind these biological processes, it is important to examine the roles of the proteins responsible for driving cell migration. Here, we outline a protocol to study the mechanisms of cell migration using the epithelial cell line (MDCK), and a three-dimensional, fibrous, self-polymerizing matrix as a model system. This optically clear extracellular matrix is easily amenable to live-cell imaging studies and better mimics the physiological, soft tissue environment. This report demonstrates a technique for directly visualizing protein localization and dynamics, and deformation of the surrounding three-dimensional matrix. Examination of protein localization and dynamics during cellular processes provides key insight into protein functions. Genetically encoded fluorescent tags provide a unique method for observing protein localization and dynamics. Using this technique, we can analyze the subcellular accumulation of key, force-generating cytoskeletal components in real-time as the cell maneuvers through the matrix. In addition, using multiple fluorescent tags with different wavelengths, we can examine the localization of multiple proteins simultaneously, thus allowing us to test, for example, whether different proteins have similar or divergent roles. Furthermore, the dynamics of fluorescently tagged proteins can be quantified using Fluorescent Recovery After Photobleaching (FRAP) analysis. This measurement assays the protein mobility and how stably bound the proteins are to the cytoskeletal network. By combining live-cell imaging with the treatment of protein function inhibitors, we can examine in real-time the changes in the distribution of proteins and morphology of migrating cells. Furthermore, we also combine live-cell imaging with the use of fluorescent tracer particles embedded within the matrix to visualize the matrix deformation during cell migration. Thus, we can visualize how a migrating cell distributes force-generating proteins, and where the traction forces are exerted to the surrounding matrix. Through these techniques, we can gain valuable insight into the roles of specific proteins and their contributions to the mechanisms of cell migration.

Mass flow and velocity profiles in Neurospora hyphae: partial plug flow dominates intra-hyphal transport.

PubMed

Abadeh, Aryan; Lew, Roger R

2013-11-01

Movement of nuclei, mitochondria and vacuoles through hyphal trunks of Neurospora crassa were vector-mapped using fluorescent markers and green fluorescent protein tags. The vectorial movements of all three were strongly correlated, indicating the central role of mass (bulk) flow in cytoplasm movements in N. crassa. Profiles of velocity versus distance from the hyphal wall did not match the parabolic shape predicted by the ideal Hagen-Poiseuille model of flow at low Reynolds number. Instead, the profiles were flat, consistent with a model of partial plug flow due to the high concentration of organelles in the flowing cytosol. The intra-hyphal pressure gradients were manipulated by localized external osmotic treatments to demonstrate the dependence of velocity (and direction) on pressure gradients within the hyphae. The data support the concept that mass transport, driven by pressure gradients, dominates intra-hyphal transport. The transport occurs by partial plug flow due to the organelles in the cytosol.

Visible and NIR spectral band combination to produce high security ID tags for automatic identification

NASA Astrophysics Data System (ADS)

Pérez-Cabré, Elisabet; Millán, María S.; Javidi, Bahram

2006-09-01

Verification of a piece of information and/or authentication of a given object or person are common operations carried out by automatic security systems that can be applied, for instance, to control the entrance to restricted areas, access to public buildings, identification of cardholders, etc. Vulnerability of such security systems may depend on the ease of counterfeiting the information used as a piece of identification for verification and authentication. To protect data against tampering, the signature that identifies an object is usually encrypted to avoid an easy recognition at human sight and an easy reproduction using conventional devices for imaging or scanning. To make counterfeiting even more difficult, we propose to combine data from visible and near infrared (NIR) spectral bands. By doing this, neither the visible content nor the NIR data by theirselves are sufficient to allow the signature recognition and thus, the identification of a given object. Only the appropriate combination of both signals permits a satisfactory authentication. In addition, the resulting signature is encrypted following a fully-phase encryption technique and the obtained complex-amplitude distribution is encoded on an ID tag. Spatial multiplexing of the encrypted signature allows us to build a distortion-invariant ID tag, so that remote authentication can be achieved even if the tag is captured under rotation or at different distances. We also explore the possibility of using partial information of the encrypted signature to simplify the ID tag design.

Effect of Starvation and the Viable-but-Nonculturable State on Green Fluorescent Protein (GFP) Fluorescence in GFP-Tagged Pseudomonas fluorescens A506

PubMed Central

Lowder, M.; Unge, A.; Maraha, N.; Jansson, J. K.; Swiggett, J.; Oliver, J. D.

2000-01-01

The green fluorescent protein (GFP) gene, gfp, of the jellyfish Aequorea victoria is being used as a reporter system for gene expression and as a marker for tracking prokaryotes and eukaryotes. Cells that have been genetically altered with the gfp gene produce a protein that fluoresces when it is excited by UV light. This unique phenotype allows gfp-tagged cells to be specifically monitored by nondestructive means. In this study we determined whether a gfp-tagged strain of Pseudomonas fluorescens continued to fluoresce under conditions under which the cells were starved, viable but nonculturable (VBNC), or dead. Epifluorescent microscopy, flow cytometry, and spectrofluorometry were used to measure fluorescence intensity in starved, VBNC, and dead or dying cells. Results obtained by using flow cytometry indicated that microcosms containing VBNC cells, which were obtained by incubation under stress conditions (starvation at 37.5°C), fluoresced at an intensity that was at least 80% of the intensity of nonstressed cultures. Similarly, microcosms containing starved cells incubated at 5 and 30°C had fluorescence intensities that were 90 to 110% of the intensity of nonstressed cells. VBNC cells remained fluorescent during the entire 6-month incubation period. In addition, cells starved at 5 or 30°C remained fluorescent for at least 11 months. Treatment of the cells with UV light or incubation at 39 or 50°C resulted in a loss of GFP from the cells. There was a strong correlation between cell death and leakage of GFP from the cells, although the extent of leakage varied depending on the treatment. Most dead cells were not GFP fluorescent, but a small proportion of the dead cells retained some GFP at a lower concentration than the concentration in live cells. Our results suggest that gfp-tagged cells remain fluorescent following starvation and entry into the VBNC state but that fluorescence is lost when the cells die, presumably because membrane integrity is lost. PMID:10919764

Dynamic MRI of Grid-Tagged Hyperpolarized Helium-3 for the Assessment of Lung Motion During Breathing

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

Cai Jing; Sheng Ke; Benedict, Stanley H.

2009-09-01

Purpose: To develop a dynamic magnetic resonance imaging (MRI) tagging technique using hyperpolarized helium-3 (HP He-3) to track lung motion. Methods and Materials: An accelerated non-Cartesian k-space trajectory was used to gain acquisition speed, at the cost of introducing image artifacts, providing a viable strategy for obtaining whole-lung coverage with adequate temporal resolution. Multiple-slice two-dimensional dynamic images of the lung were obtained in three healthy subjects after inhaling He-3 gas polarized to 35%-40%. Displacement, strain, and ventilation maps were computed from the observed motion of the grid peaks. Results: Both temporal and spatial variations of pulmonary mechanics were observed inmore » normal subjects, including shear motion between different lobes of the same lung. Conclusion: These initial results suggest that dynamic imaging of grid-tagged hyperpolarized magnetization may potentially be a powerful tool for observing and quantifying pulmonary biomechanics on a regional basis and for assessing, validating, and improving lung deformable image registration algorithms.« less

Nonlethal gill biopsy does not affect juvenile chinook salmon implanted with radio transmitters

USGS Publications Warehouse

Martinelli-Liedtke, T. L.; Shively, R.S.; Holmberg, G.S.; Sheer, M.B.; Schrock, R.M.

1999-01-01

Using gastric and surgical transmitter implantation, we compared radio-tagged juvenile chinook salmon Oncorhynchus tshawytscha (T(O)) with tagged fish also having a gill biopsy (T(B)) to determine biopsy effects on fish implanted with radio transmitters. We found no evidence during the 21-d period to suggest that a gill biopsy reduced survival, growth, or gross condition of the tagged-biopsy group, regardless of transmitter implantation technique. We recorded 100% survival of all treatment groups. Relative growth rates of T(O) and T(B) fish did not differ significantly. Leukocrit and lysozyme levels were not significantly different among groups, suggesting that no signs of infection were present. Our findings suggest that small chinook salmon can tolerate the combination of transmitter implantation and gill biopsy without compromising condition relative to fish receiving only the transmitter. We believe a gill biopsy can be used in field telemetry studies, especially when physiological data are needed in addition to behavioral data.

Determining composition of micron-scale protein deposits in neurodegenerative disease by spatially targeted optical microproteomics.

PubMed

Hadley, Kevin C; Rakhit, Rishi; Guo, Hongbo; Sun, Yulong; Jonkman, James E N; McLaurin, Joanne; Hazrati, Lili-Naz; Emili, Andrew; Chakrabartty, Avijit

2015-09-29

Spatially targeted optical microproteomics (STOMP) is a novel proteomics technique for interrogating micron-scale regions of interest (ROIs) in mammalian tissue, with no requirement for genetic manipulation. Methanol or formalin-fixed specimens are stained with fluorescent dyes or antibodies to visualize ROIs, then soaked in solutions containing the photo-tag: 4-benzoylbenzyl-glycyl-hexahistidine. Confocal imaging along with two photon excitation are used to covalently couple photo-tags to all proteins within each ROI, to a resolution of 0.67 µm in the xy-plane and 1.48 µm axially. After tissue solubilization, photo-tagged proteins are isolated and identified by mass spectrometry. As a test case, we examined amyloid plaques in an Alzheimer's disease (AD) mouse model and a post-mortem AD case, confirming known plaque constituents and discovering new ones. STOMP can be applied to various biological samples including cell lines, primary cell cultures, ex vivo specimens, biopsy samples, and fixed post-mortem tissue.

Learning Semantic Tags from Big Data for Clinical Text Representation.

PubMed

Li, Yanpeng; Liu, Hongfang

2015-01-01

In clinical text mining, it is one of the biggest challenges to represent medical terminologies and n-gram terms in sparse medical reports using either supervised or unsupervised methods. Addressing this issue, we propose a novel method for word and n-gram representation at semantic level. We first represent each word by its distance with a set of reference features calculated by reference distance estimator (RDE) learned from labeled and unlabeled data, and then generate new features using simple techniques of discretization, random sampling and merging. The new features are a set of binary rules that can be interpreted as semantic tags derived from word and n-grams. We show that the new features significantly outperform classical bag-of-words and n-grams in the task of heart disease risk factor extraction in i2b2 2014 challenge. It is promising to see that semantics tags can be used to replace the original text entirely with even better prediction performance as well as derive new rules beyond lexical level.

An internal affinity-tag for purification and crystallization of the siderophore receptor FhuA, integral outer membrane protein from Escherichia coli K-12.

PubMed

Ferguson, A D; Breed, J; Diederichs, K; Welte, W; Coulton, J W

1998-07-01

FhuA (Mr 78,992, 714 amino acids), siderophore receptor for ferrichrome-iron in the outer membrane of Escherichia coli, was affinity tagged, rapidly purified, and crystallized. To obtain FhuA in quantities sufficient for crystallization, a hexahistidine tag was genetically inserted into the fhuA gene after amino acid 405, which resides in a known surface-exposed loop. Recombinant FhuA405.H6 was overexpressed in an E. coli strain that is devoid of several major porins and using metal-chelate chromatography was purified in large amounts to homogeneity. FhuA crystals were grown using the hanging drop vapor diffusion technique and were suitable for X-ray diffraction analysis. On a rotating anode X-ray source, diffraction was observed to 3.0 A resolution. The crystals belong to space group P6(1) or P6(5) with unit cell dimensions of a=b=174 A, c=88 A (alpha=beta=90 degrees, gamma=120 degrees).

Recovery of coded wire tags at a caspian tern colony in San Francisco Bay: A technique to evaluate impacts of avian predation on juvenile salmonids

USGS Publications Warehouse

Evans, A.F.; Roby, D.D.; Collis, K.; Cramer, B.M.; Sheggeby, J.A.; Adrean, L.J.; Battaglia, D.S.; Lyons, Donald E.

2011-01-01

We recovered coded wire tags (CWTs) from a colony of Caspian terns Hydroprogne caspia on Brooks Island in San Francisco Bay, California, to evaluate predation on juvenile salmonids originating from the Sacramento and San Joaquin rivers. Subsamples of colony substrate representing 11.7% of the nesting habitat used by the terns yielded 2,079 salmonid CWTs from fish released and subsequently consumed by terns in 2008. The estimated number of CWTs deposited on the entire tern colony was 40,143 (ranging from 26,763 to 80,288), once adjustments were made to account for tag loss and the total amount of nesting habitat used by terns. Tags ingested by terns and then egested on the colony were undamaged, and the tags' complete numeric codes were still identifiable. The CWTs found on the tern colony indicated that hatchery Chinook salmon Oncorhynchus tshawytscha trucked to and released in San Pablo Bay were significantly more likely to be consumed by Caspian terns than Chinook salmon that migrated in-river to the bay; 99.7% of all tags recovered were from bay-released Chinook salmon. Of the CWTs recovered on the tern colony, 98.0% were from fall-run Chinook salmon, indicating a higher susceptibility to tern predation than for the spring run type. None of the approximately 518,000 wild Chinook salmon that were coded-wire-tagged and released in the basin were recovered on the tern colony, suggesting that the impacts on wild, U.S. Endangered Species Act-listed Chinook salmon populations were minimal in 2008. Overall, we estimate that 0.3% of the approximately 12.3 million coded-wire-tagged Chinook salmon released in the basin in 2008 were subsequently consumed by Caspian terns from the Brooks Island colony. These results indicate that CWTs implanted in juvenile salmon can be recovered from a piscivorous waterbird colony and used to evaluate smolt losses for runs that are tagged. Abstract We recovered coded wire tags (CWTs) from a colony of Caspian terns Hydroprogne caspia on Brooks Island in San Francisco Bay, California, to evaluate predation on juvenile salmonids originating from the Sacramento and San Joaquin rivers. Subsamples of colony substrate representing 11.7% of the nesting habitat used by the terns yielded 2,079 salmonid CWTs from fish released and subsequently consumed by terns in 2008. The estimated number of CWTs deposited on the entire tern colony was 40,143 (ranging from 26,763 to 80,288), once adjustments were made to account for tag loss and the total amount of nesting habitat used by terns. Tags ingested by terns and then egested on the colony were undamaged, and the tags' complete numeric codes were still identifiable. The CWTs found on the tern colony indicated that hatchery Chinook salmon Oncorhynchus tshawytscha trucked to and released in San Pablo Bay were significantly more likely to be consumed by Caspian terns than Chinook salmon that migrated in-river to the bay; 99.7% of all tags recovered were from bay-released Chinook salmon. Of the CWTs recovered on the tern colony, 98.0% were from fall-run Chinook salmon, indicating a higher susceptibility to tern predation than for the spring run type. None of the approximately 518,000 wild Chinook salmon that were coded-wire-tagged and released in the basin were recovered on the tern colony, suggesting that the impacts on wild, U.S. Endangered Species Act-listed Chinook salmon populations were minimal in 2008. Overall, we estimate that 0.3% of the approximately 12.3 million coded-wire-tagged Chinook salmon released in the basin in 2008 were subsequently consumed by Caspian terns from the Brooks Island colony. These results indicate that CWTs implanted in juvenile salmon can be recovered from a piscivorous waterbird colony and used to evaluate smolt losses for runs that are tagged ?? American Fisheries Society 2011.

Development of expressed sequence tag-simple sequence repeat markers for genetic characterization and population structure analysis of Praxelis clematidea (Asteraceae).

PubMed

Wang, Q Z; Huang, M; Downie, S R; Chen, Z X

2016-05-23

Invasive plants tend to spread aggressively in new habitats and an understanding of their genetic diversity and population structure is useful for their management. In this study, expressed sequence tag-simple sequence repeat (EST-SSR) markers were developed for the invasive plant species Praxelis clematidea (Asteraceae) from 5548 Stevia rebaudiana (Asteraceae) expressed sequence tags (ESTs). A total of 133 microsatellite-containing ESTs (2.4%) were identified, of which 56 (42.1%) were hexanucleotide repeat motifs and 50 (37.6%) were trinucleotide repeat motifs. Of the 24 primer pairs designed from these 133 ESTs, 7 (29.2%) resulted in significant polymorphisms. The number of alleles per locus ranged from 5 to 9. The relatively high genetic diversity (H = 0.2667, I = 0.4212, and P = 100%) of P. clematidea was related to high gene flow (Nm = 1.4996) among populations. The coefficient of population differentiation (GST = 0.2500) indicated that most genetic variation occurred within populations. A Mantel test suggested that there was significant correlation between genetic distance and geographical distribution (r = 0.3192, P = 0.012). These results further support the transferability of EST-SSR markers between closely related genera of the same family.

A method for the analysis of sugars in biological systems using reductive amination in combination with hydrophilic interaction chromatography and high resolution mass spectrometry.

PubMed

Bawazeer, Sami; Muhsen Ali, Ali; Alhawiti, Aliyah; Khalaf, Abedawn; Gibson, Colin; Tusiimire, Jonans; Watson, David G

2017-05-01

Separation of sugar isomers extracted from biological samples is challenging because of their natural occurrence as alpha and beta anomers and, in the case of hexoses, in their pyranose and furanose forms. A reductive amination method was developed for the tagging of sugars with the aim of it becoming part of a metabolomics work flow. The best separation of the common hexoses (glucose, fructose, mannose and galactose) was achieved when 2 H 5 -aniline was used as the tagging reagent in combination with separation on a ZICHILIC column. The method was used to tag a range of sugars including pentoses and uronic acids. The method was simple to perform and was able to improve both the separation of sugars and their response to electrospray ionisation. The method was applied to the profiling of sugars in urine where a number of hexose and pentose isomers could be observed. It was also applied to the quantification of sugars in post-mortem brain samples from three control samples and three samples from individuals who had suffered from bipolar disorder. Copyright © 2017 Elsevier B.V. All rights reserved.

Large-scale production of (GeTe) x (AgSbTe 2) 100$-$x (x=75, 80, 85, 90) with enhanced thermoelectric properties via gas-atomization and spark plasma sintering

DOE PAGES

Kim, Hyo-Seob; Ames Lab., Ames, IA; Dharmaiah, Peyala; ...

2017-01-30

(GeTe) x(AgSbTe 2) 100$-$x: TAGS thermoelectrics are an attractive class of materials due to their combination of non-toxicity and good conversion efficiency at mid-temperature ranges. Here in the present work, we have utilized energy and time efficient high-pressure gas atomization and spark-plasma sintering techniques for large-scale preparation of samples with varying composition (i.e., (GeTe) x(AgSbTe 2) 100$-$x where x = 75, 80, 85, and 90). High-temperature x-ray diffraction was used to understand the phase transformation mechanism of the as-atomized powders. Detailed high-resolution transmission electron microscopy of the sintered samples revealed the presence of nanoscale precipitates, antiphase, and twin boundaries. Themore » nanoscale twins and antiphase boundaries serve as phonon scattering centers, leading to the reduction of total thermal conductivity in TAGS-80 and 90 samples. The maximum ZT obtained was 1.56 at 623 K for TAGS-90, which was ~94% improvement compared to values previously reported. The presence of the twin boundaries also resulted in a high fracture toughness (K IC) of the TAGS-90 sample due to inhibition of dislocation movement at the twin boundary.« less

Population assessment of tropical tuna based on their associative behavior around floating objects.

PubMed

Capello, M; Deneubourg, J L; Robert, M; Holland, K N; Schaefer, K M; Dagorn, L

2016-11-03

Estimating the abundance of pelagic fish species is a challenging task, due to their vast and remote habitat. Despite the development of satellite, archival and acoustic tagging techniques that allow the tracking of marine animals in their natural environments, these technologies have so far been underutilized in developing abundance estimations. We developed a new method for estimating the abundance of tropical tuna that employs these technologies and exploits the aggregative behavior of tuna around floating objects (FADs). We provided estimates of abundance indices based on a simulated set of tagged fish and studied the sensitivity of our method to different association dynamics, FAD numbers, population sizes and heterogeneities of the FAD-array. Taking the case study of yellowfin tuna (Thunnus albacares) acoustically-tagged in Hawaii, we implemented our approach on field data and derived for the first time the ratio between the associated and the total population. With more extensive and long-term monitoring of FAD-associated tunas and good estimates of the numbers of fish at FADs, our method could provide fisheries-independent estimates of populations of tropical tuna. The same approach can be applied to obtain population assessments for any marine and terrestrial species that display associative behavior and from which behavioral data have been acquired using acoustic, archival or satellite tags.

Single tag for total carbohydrate analysis.

PubMed

Anumula, Kalyan Rao

2014-07-15

Anthranilic acid (2-aminobenzoic acid, 2-AA) has the remarkable property of reacting rapidly with every type of reducing carbohydrate. Reactivity of 2-AA with carbohydrates in aqueous solutions surpasses all other tags reported to date. This unique capability is attributed to the strategically located -COOH which accelerates Schiff base formation. Monosaccharides, oligosaccharides (N-, O-, and lipid linked and glycans in secretory fluids), glycosaminoglycans, and polysaccharides can be easily labeled with 2-AA. With 2-AA, labeling is simple in aqueous solutions containing proteins, peptides, buffer salts, and other ingredients (e.g., PNGase F, glycosidase, and transferase reaction mixtures). In contrast, other tags require relatively pure glycans for labeling in anhydrous dimethyl sulfoxide-acetic acid medium. Acidic conditions are known to cause desialylation, thus requiring a great deal of attention to sample preparation. Simpler labeling is achieved with 2-AA within 30-60 min in mild acetate-borate buffered solution. 2-AA provides the highest sensitivity and resolution in chromatographic methods for carbohydrate analysis in a simple manner. Additionally, 2-AA is uniquely qualified for quantitative analysis by mass spectrometry in the negative mode. Analyses of 2-AA-labeled carbohydrates by electrophoresis and other techniques have been reported. Examples cited here demonstrate that 2-AA is the universal tag for total carbohydrate analysis. Copyright © 2014 Elsevier Inc. All rights reserved.

Click-electron microscopy for imaging metabolically tagged non-protein biomolecules

PubMed Central

Ngo, John T.; Adams, Stephen R.; Deerinck, Thomas J.; Boassa, Daniela; Rodriguez-Rivera, Frances; Palida, Sakina F.; Bertozzi, Carolyn R.; Ellisman, Mark H.; Tsien, Roger Y.

2016-01-01

Electron microscopy (EM) has long been the main technique to image cell structures with nanometer resolution, but has lagged behind light microscopy in the crucial ability to make specific molecules stand out. Here we introduce “Click-EM,” a labeling technique for correlative light microscopy and EM imaging of non-protein biomolecules. In this approach, metabolic labeling substrates containing bioorthogonal functional groups are provided to cells for incorporation into biopolymers by endogenous biosynthetic machinery. The unique chemical functionality of these analogs is exploited for selective attachment of singlet oxygen-generating fluorescent dyes via bioorthogonal “click chemistry” ligations. Illumination of dye-labeled structures generates singlet oxygen to locally catalyze the polymerization of diaminobenzidine into an osmiophilic reaction product that is readily imaged by EM. We describe the application of Click-EM in imaging metabolically tagged DNA, RNA, and lipids in cultured cells and neurons, and highlight its use in tracking peptidoglycan synthesis in the Gram-positive bacterium Listeria monocytogenes. PMID:27110681

Instability in extensional microflow of aqueous gel

NASA Astrophysics Data System (ADS)

Bryce, Robert; Freeman, Mark

2007-03-01

Microfluidic devices are typically characterized by laminar flows, often leading to diffusion limited mixing. Recently it has been demonstrated that the addition of polymer to fluids can lead to elastic instabilities and, under some conditions, turbulence at arbitrarily low Reynolds numbers in mechanically driven flows [1]. We investigated electroosmotic driven extensional flow of an aqueous polymer gel. Microchannels with 100 micron width and 20 micron depth with the characteristic ``D'' chemical etch cross section were formed in glass. A Y-channel geometry with two input channels and a single output created extensional flow at the channel intersection. Instabilities where observed in the extensional region by fluorescently tagging one input stream. Instabilities were characterized by 1/f spectra in laser induced fluorescent brightness profiles. Due to the simple geometry of extensional flow and the importance of electroosmotic flows for integrated applications and in scaling, this is of interest for device applications. [1] A. Groisman and V. Steinberg, Nature 405, 53-55, 2000.

Feasibility of Quantifying Arterial Cerebral Blood Volume Using Multiphase Alternate Ascending/Descending Directional Navigation (ALADDIN).

PubMed

Kim, Ki Hwan; Choi, Seung Hong; Park, Sung-Hong

2016-01-01

Arterial cerebral blood volume (aCBV) is associated with many physiologic and pathologic conditions. Recently, multiphase balanced steady state free precession (bSSFP) readout was introduced to measure labeled blood signals in the arterial compartment, based on the fact that signal difference between labeled and unlabeled blood decreases with the number of RF pulses that is affected by blood velocity. In this study, we evaluated the feasibility of a new 2D inter-slice bSSFP-based arterial spin labeling (ASL) technique termed, alternate ascending/descending directional navigation (ALADDIN), to quantify aCBV using multiphase acquisition in six healthy subjects. A new kinetic model considering bSSFP RF perturbations was proposed to describe the multiphase data and thus to quantify aCBV. Since the inter-slice time delay (TD) and gap affected the distribution of labeled blood spins in the arterial and tissue compartments, we performed the experiments with two TDs (0 and 500 ms) and two gaps (300% and 450% of slice thickness) to evaluate their roles in quantifying aCBV. Comparison studies using our technique and an existing method termed arterial volume using arterial spin tagging (AVAST) were also separately performed in five subjects. At 300% gap or 500-ms TD, significant tissue perfusion signals were demonstrated, while tissue perfusion signals were minimized and arterial signals were maximized at 450% gap and 0-ms TD. ALADDIN has an advantage of visualizing bi-directional flow effects (ascending/descending) in a single experiment. Labeling efficiency (α) of inter-slice blood flow effects could be measured in the superior sagittal sinus (SSS) (20.8±3.7%.) and was used for aCBV quantification. As a result of fitting to the proposed model, aCBV values in gray matter (1.4-2.3 mL/100 mL) were in good agreement with those from literature. Our technique showed high correlation with AVAST, especially when arterial signals were accentuated (i.e., when TD = 0 ms) (r = 0.53). The bi-directional perfusion imaging with multiphase ALADDIN approach can be an alternative to existing techniques for quantification of aCBV.

Redox active polymer devices and methods of using and manufacturing the same

DOEpatents

Johnson, Paul; Bautista-Martinez, Jose Antonio; Friesen, Cody; Switzer, Elise

2018-06-05

The disclosed technology relates generally to apparatus comprising conductive polymers and more particularly to tag and tag devices comprising a redox-active polymer film, and method of using and manufacturing the same. In one aspect, an apparatus includes a substrate and a conductive structure formed on the substrate which includes a layer of redox-active polymer film having mobile ions and electrons. The conductive structure further includes a first terminal and a second terminal configured to receive an electrical signal therebetween, where the layer of redox-active polymer is configured to conduct an electrical current generated by the mobile ions and the electrons in response to the electrical signal. The apparatus additionally includes a detection circuit operatively coupled to the conductive structure and configured to detect the electrical current flowing through the conductive structure.

Development of Ciprofloxacin-loaded contact lenses using fluorous chemistry

PubMed Central

Zhu, Zhiling; Li, Siheng; McDermott, Alison M.

2017-01-01

In this work, we developed a simple method to load drugs into commercially available contact lenses utilizing fluorous chemistry. We demonstrated this method using model compounds including fluorous-tagged fluorescein and antibiotic ciprofloxacin. We showed that fluorous interactions facilitated the loading of model molecules into fluorocarbon-containing contact lenses, and that the release profiles exhibited sustained release. Contact lenses loaded with fluorous-tagged ciprofloxacin exhibited antimicrobial activity against Pseudomonas aeruginosa in vitro, while no cytotoxicity towards human corneal epithelial cells was observed. To mimic the tear turnover, we designed a porcine eye infection model under flow conditions. Significantly, the modified lenses also exhibited antimicrobial efficacy against Pseudomonas aeruginosa in the ex vivo infection model. Overall, utilizing fluorous chemistry, we can construct a drug delivery system that exhibits high drug loading capacity, sustained drug release, and robust biological activity. PMID:28188995

Assimilable organic carbon (AOC) determination using GFP-tagged Pseudomonas fluorescens P-17 in water by flow cytometry.

PubMed

Tang, Peng; Wu, Jie; Liu, Hou; Liu, Youcai; Zhou, Xingding

2018-01-01

One of the newly developed methods for Assimilable organic carbon (AOC) determination is leveraged on the cell enumeration by flow cytometry (FC) which could provide a rapid and automated solution for AOC measurement. However, cell samples staining with fluorescence dye is indispensable to reduce background and machine noise. This step would bring additional cost and time consuming for this method. In this study, a green fluorescence protein (GFP) tagged strain derived of AOC testing strain Pseudomonas fluorescens P-17 (GFP-P17) was generated using Tn5 transposon mutagenesis. Continuous culture of this mutant GFP-P17 showed stable expression of eGFP signal detected by flow cytometry without staining step. In addition, this GFP-P17 strain displayed faster growth rate and had a wider range of carbon substrate utilization patterns as compared with P17 wild-type. With this strain, the capability of a new FC method with no dye staining was explored in standard acetate solution, which suggests linear correlation of counts with acetate carbon concentration. Furthermore, this FC method with GFP-P17 strain is applicable in monitoring GAC/BAC efficiency and condition as similar trends of AOC level in water treatment process were measured by both FC method and conventional spread plating count method. Therefore, this fast and easily applicable GFP-P17 based FC method could serve as a tool for routine microbiological drinking water monitoring.

Detecting relay attacks on RFID communication systems using quantum bits

NASA Astrophysics Data System (ADS)

Jannati, Hoda; Ardeshir-Larijani, Ebrahim

2016-11-01

RFID systems became widespread in variety of applications because of their simplicity in manufacturing and usability. In the province of critical infrastructure protection, RFID systems are usually employed to identify and track people, objects and vehicles that enter restricted areas. The most important vulnerability which is prevalent among all protocols employed in RFID systems is against relay attacks. Until now, to protect RFID systems against this kind of attack, the only approach is the utilization of distance-bounding protocols which are not applicable over low-cost devices such as RFID passive tags. This work presents a novel technique using emerging quantum technologies to detect relay attacks on RFID systems. Recently, it is demonstrated that quantum key distribution (QKD) can be implemented in a client-server scheme where client only requires an on-chip polarization rotator that may be integrated into a handheld device. Now we present our technique for a tag-reader scenario which needs similar resources as the mentioned QKD scheme. We argue that our technique requires less resources and provides lower probability of false alarm for the system, compared with distance-bounding protocols, and may pave the way to enhance the security of current RFID systems.

Design and development of a magnetic device for mesenchymal stem cell retaining in deep targets

NASA Astrophysics Data System (ADS)

Banis, G. C.

2017-12-01

This paper focuses on the retaining of mesenchymal stem cells in blood flow conditions using the appropriate magnetic field. Mesenchymal stem cells can be tagged with magnetic nanoparticles and thus, they can be manipulated from distance, through the application of an external magnetic field. In this paper the case of kidney as target of the therapy is being studied.

Adaptive Meshing of Ship Air-Wake Flowfields

DTIC Science & Technology

2014-10-21

performs cut- cell operations at geometry boundaries. A second-order spatial finite-volume scheme has been incorporated with explicit first order...The cells intersected by the geometry are handled using the “cut- cell ” approach, which is basically creating arbitrary polyhedral elements with...appropriate surface boundary conditions. Any cells completely outside the computational domain are tagged external and not solved in the flow solution

Live-cell stimulated Raman scattering imaging of alkyne-tagged biomolecules.

PubMed

Hong, Senlian; Chen, Tao; Zhu, Yuntao; Li, Ang; Huang, Yanyi; Chen, Xing

2014-06-02

Alkynes can be metabolically incorporated into biomolecules including nucleic acids, proteins, lipids, and glycans. In addition to the clickable chemical reactivity, alkynes possess a unique Raman scattering within the Raman-silent region of a cell. Coupling this spectroscopic signature with Raman microscopy yields a new imaging modality beyond fluorescence and label-free microscopies. The bioorthogonal Raman imaging of various biomolecules tagged with an alkyne by a state-of-the-art Raman imaging technique, stimulated Raman scattering (SRS) microscopy, is reported. This imaging method affords non-invasiveness, high sensitivity, and molecular specificity and therefore should find broad applications in live-cell imaging. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A Concealed Barcode Identification System Using Terahertz Time-domain Spectroscopy

NASA Astrophysics Data System (ADS)

Guan, Yu; Yamamoto, Manabu; Kitazawa, Toshiyuki; Tripathi, Saroj R.; Takeya, Kei; Kawase, Kodo

2015-03-01

We present a concealed terahertz barcode/chipless tag to achieve remote identification through an obstructing material using terahertz radiation. We show scanned terahertz reflection spectral images of barcodes concealed by a thick obstacle. A concealed and double- side printed terahertz barcode structure is proposed, and we demonstrate that our design has better performance in definition than a single-side printed barcode using terahertz time-domain spectroscopy. This technique combines the benefits of a chipless tag to read encoded information covered by an optically opaque material with low cost and a simple fabrication process. Simulations are also described, along with an explanation of the principle of the terahertz barcode identification system.

A Vision-Based System for Object Identification and Information Retrieval in a Smart Home

NASA Astrophysics Data System (ADS)

Grech, Raphael; Monekosso, Dorothy; de Jager, Deon; Remagnino, Paolo

This paper describes a hand held device developed to assist people to locate and retrieve information about objects in a home. The system developed is a standalone device to assist persons with memory impairments such as people suffering from Alzheimer's disease. A second application is object detection and localization for a mobile robot operating in an ambient assisted living environment. The device relies on computer vision techniques to locate a tagged object situated in the environment. The tag is a 2D color printed pattern with a detection range and a field of view such that the user may point from a distance of over 1 meter.

Screen printed UHF antennas on flexible substrates

NASA Astrophysics Data System (ADS)

Janeczek, Kamil; Młożniak, Anna; Kozioł, Grażyna; Araźna, Aneta; Jakubowska, Małgorzata; Bajurko, Paweł

2010-09-01

Printed electronics belongs to the most important developing electronics technologies. It provides new possibilities to produce low cost and large area devices. In its range several applications can be distinguished like printed batteries, OLED, biosensors, photovoltaic cells or RFID tags. In the presented investigation, antennas working in UHF frequency range were elaborated. It can be applied in the future for flexible RFID tags. To produce these antennas polymer paste with silver flakes was used. It was deposited on two flexible substrates (foil and photo paper) with screen printing techniques. After printing process surface profile, electrical and microwave parameters of performed antennas were measured using digital multimeter and network analyzer, relatively. Furthermore, a thickness of printed layers was measured.

Fluorous Parallel Synthesis of A Hydantoin/Thiohydantoin Library

PubMed Central

Lu, Yimin; Zhang, Wei

2007-01-01

Fluorous tagging strategy is applied to solution-phase parallel synthesis of a library containing hydantoin and thiohydantoin analogs. Two perfluoroalkyl (Rf)-tagged α-amino esters each react with 6 aromatic aldehydes under reductive amination conditions. Twelve amino esters then each react with 10 isocyanates and isothiocyanates in parallel. The resulting 120 ureas and thioureas undergo spontaneous cyclization to form the corresponding hydantoins and thiohydantoins. The intermediate and final product purifications are performed with solid-phase extraction (SPE) over FluoroFlash™ cartridges, no chromatography is required. Using standard instruments and straightforward SPE technique, one chemist accomplished the 120-member library synthesis in less than 5 working days, including starting material synthesis and product analysis. PMID:15789556

Computational design optimization for microfluidic magnetophoresis

PubMed Central

Plouffe, Brian D.; Lewis, Laura H.; Murthy, Shashi K.

2011-01-01

Current macro- and microfluidic approaches for the isolation of mammalian cells are limited in both efficiency and purity. In order to design a robust platform for the enumeration of a target cell population, high collection efficiencies are required. Additionally, the ability to isolate pure populations with minimal biological perturbation and efficient off-chip recovery will enable subcellular analyses of these cells for applications in personalized medicine. Here, a rational design approach for a simple and efficient device that isolates target cell populations via magnetic tagging is presented. In this work, two magnetophoretic microfluidic device designs are described, with optimized dimensions and operating conditions determined from a force balance equation that considers two dominant and opposing driving forces exerted on a magnetic-particle-tagged cell, namely, magnetic and viscous drag. Quantitative design criteria for an electromagnetic field displacement-based approach are presented, wherein target cells labeled with commercial magnetic microparticles flowing in a central sample stream are shifted laterally into a collection stream. Furthermore, the final device design is constrained to fit on standard rectangular glass coverslip (60 (L)×24 (W)×0.15 (H) mm3) to accommodate small sample volume and point-of-care design considerations. The anticipated performance of the device is examined via a parametric analysis of several key variables within the model. It is observed that minimal currents (<500 mA) are required to generate magnetic fields sufficient to separate cells from the sample streams flowing at rate as high as 7 ml∕h, comparable to the performance of current state-of-the-art magnet-activated cell sorting systems currently used in clinical settings. Experimental validation of the presented model illustrates that a device designed according to the derived rational optimization can effectively isolate (∼100%) a magnetic-particle-tagged cell population from a homogeneous suspension even in a low abundance. Overall, this design analysis provides a rational basis to select the operating conditions, including chamber and wire geometry, flow rates, and applied currents, for a magnetic-microfluidic cell separation device. PMID:21526007

An optical clearing technique for plant tissues allowing deep imaging and compatible with fluorescence microscopy.

PubMed

Warner, Cherish A; Biedrzycki, Meredith L; Jacobs, Samuel S; Wisser, Randall J; Caplan, Jeffrey L; Sherrier, D Janine

2014-12-01

We report on a nondestructive clearing technique that enhances transmission of light through specimens from diverse plant species, opening unique opportunities for microscope-enabled plant research. After clearing, plant organs and thick tissue sections are amenable to deep imaging. The clearing method is compatible with immunocytochemistry techniques and can be used in concert with common fluorescent probes, including widely adopted protein tags such as GFP, which has fluorescence that is preserved during the clearing process. © 2014 American Society of Plant Biologists. All Rights Reserved.

Webcam-based flow cytometer using wide-field imaging for low cell number detection at high throughput.

PubMed

Balsam, Joshua; Bruck, Hugh Alan; Rasooly, Avraham

2014-09-07

Here we describe a novel low-cost flow cytometer based on a webcam capable of low cell number detection in a large volume which may overcome the limitations of current flow cytometry. Several key elements have been combined to yield both high throughput and high sensitivity. The first element is a commercially available webcam capable of 187 frames per second video capture at a resolution of 320 × 240 pixels. The second element in this design is a 1 W 450 nm laser module for area-excitation, which combined with the webcam allows for rapid interrogation of a flow field. The final element is a 2D flow-cell which overcomes the flow limitation of hydrodynamic focusing and allows for higher sample throughput in a wider flow field. This cell allows for the linear velocity of target cells to be lower than in a conventional "1D" hydrodynamic focusing flow-cells typically used in cytometry at similar volumetric flow rates. It also allows cells to be imaged at the full frame rate of the webcam. Using this webcam-based flow cytometer with wide-field imaging, it was confirmed that the detection of fluorescently tagged 5 μm polystyrene beads in "1D" hydrodynamic focusing flow-cells was not practical for low cell number detection due to streaking from the motion of the beads, which did not occur with the 2D flow-cell design. The sensitivity and throughput of this webcam-based flow cytometer was then investigated using THP-1 human monocytes stained with SYTO-9 florescent dye in the 2D flow-cell. The flow cytometer was found to be capable of detecting fluorescently tagged cells at concentrations as low as 1 cell per mL at flow rates of 500 μL min(-1) in buffer and in blood. The effectiveness of detection was concentration dependent: at 100 cells per mL 84% of the cells were detected compared to microscopy, 10 cells per mL 79% detected and 1 cell per mL 59% of the cells were detected. With the blood samples spiked to 100 cells per mL, the average concentration for all samples was 91.4 cells per mL, with a 95% confidence interval of 86-97 cells per mL. These low cell concentrations and the large volume capabilities of the system may overcome the limitations of current cytometry, and are applicable to rare cell (such as circulating tumor cell) detection The simplicity and low cost of this device suggests that it may have a potential use in developing point-of-care clinical flow cytometry for resource-poor settings associated with global health.

Trip-Induced Transition Measurements in a Hypersonic Boundary Layer Using Molecular Tagging Velocimetry

NASA Technical Reports Server (NTRS)

Bathel, Brett F.; Danehy, Paul M.; Jones, Stephen B.; Johansen, Craig T.; Goyne, Christopher P.

2013-01-01

Measurements of mean streamwise velocity, fluctuating streamwise velocity, and instantaneous streamwise velocity profiles in a hypersonic boundary layer were obtained over a 10-degree half-angle wedge model. A laser-induced fluorescence-based molecular tagging velocimetry technique was used to make the measurements. The nominal edge Mach number was 4.2. Velocity profiles were measured both in an untripped boundary layer and in the wake of a 4-mm diameter cylindrical tripping element centered 75.4 mm downstream of the sharp leading edge. Three different trip heights were investigated: k = 0.53 mm, k = 1.0 mm and k = 2.0 mm. The laminar boundary layer thickness at the position of the measurements was approximately 1 mm, though the exact thickness was dependent on Reynolds number and wall temperature. All of the measurements were made starting from a streamwise location approximately 18 mm downstream of the tripping element. This measurement region continued approximately 30 mm in the streamwise direction. Additionally, measurements were made at several spanwise locations. An analysis of flow features show how the magnitude, spatial location, and spatial growth of streamwise velocity instabilities are affected by parameters such as the ratio of trip height to boundary layer thickness and roughness Reynolds number. The fluctuating component of streamwise velocity measured along the centerline of the model increased from approximately 75 m/s with no trip to +/-225 m/s with a 0.53-mm trip, and to +/-240 m/s with a 1-mm trip, while holding the freestream Reynolds number constant. These measurements were performed in the 31-inch Mach 10 Air Tunnel at the NASA Langley Research Center.

Distinct Motion of GFP-Tagged Histone Expressing Cells Under AC Electrokinetics in Electrode-Multilayered Microfluidic Device.

PubMed

Yao, Jiafeng; Sugawara, Michiko; Obara, Hiromichi; Mizutani, Takeomi; Takei, Masahiro

2017-12-01

The distinct motion of GFP-tagged histone expressing cells (Histone-GFP type cells) has been investigated under ac electrokinetics in an electrode-multilayered microfluidic device as compared with Wild type cells and GFP type cells in terms of different intracellular components. The Histone-GFP type cells were modified by the transfection of green fluorescent protein-fused histone from the human lung fibroblast cell line. The velocity of the Histone-GFP type cells obtained by particle tracking velocimetry technique is faster than Wild type cells by 24.9% and GFP type cells by 57.1%. This phenomenon is caused by the more amount of proteins in the intracellular of single Histone-GFP type cell than that of the Wild type and GFP type cells. The more amount of proteins in the Histone-GFP type cells corresponds to a lower electric permittivity ϵ c of the cells, which generates a lower dielectrophoretic force exerting on the cells. The velocity of Histone-GFP type cells is well agreed with Eulerian-Lagrangian two-phase flow simulation by 4.2% mean error, which proves that the fluid motion driven by thermal buoyancy and electrothermal force dominates the direction of cells motion, while the distinct motion of Histone-GFP type cells is caused by dielectrophoretic force. The fluid motion does not generate a distinct drag motion for Histone-GFP type cells because the Histone-GFP type cells have the same size to the Wild type and GFP type cells. These results clarified the mechanism of cells motion in terms of intracellular components, which helps to improve the cell manipulation efficiency with electrokinetics.

Fundamental Mixing and Combustion Experiments for Propelled Hypersonic Flight. Chaper 7

NASA Technical Reports Server (NTRS)

Diskin, G. S.; Danehy, P. M.; Drummond, J. P.; Cutler, A. D.

2002-01-01

The first experiment is a study of a coaxial jet discharging into stagnant laboratory air, with center jet of a mixture of 5% oxygen and 95% helium by volume and coflow jet of air. The exit flow pressure of both center-jet and coflow nozzles is 1 atmosphere. The presence of oxygen in the center jet is to allow the use of an oxygen flow-tagging technique (RELIEF4) to obtain non-intrusive velocity measurements. Both jets are nominally Mach 1.8, but, because of the greater speed of sound, the center jet velocity is more than twice that of the coflow. The mixing layer which forms between the center jet and the coflow near the nozzle exit is compressible, with a calculated convective Mach number of approximately 0.7. This geometry has several advantages: The streamwise development of the flow is generally dominated by turbulent stresses (rather than pressure forces), and thus calculations are sensitive to turbulence modeling. It includes features present in supersonic combustors, including a compressible mixing layer near the nozzle exit and a light-gas/air plume downstream. Since it is a free jet, it provides easy access for both optical instrumentation and probes. Since it is axisymmetric, it requires fewer experimental measurements to fully characterize, and calculations can be performed with more modest computer resources. However, weak shock waves formed at the nozzle exit strengthen and turn normal as they approach the axis, complicating the flow. Care is thus taken in the design of the facility to provide as near as possible to 1-D flow at the exit of both center and coflow nozzles, and to minimize the strength of waves generated at the nozzle exit. Results from this experiment are compared to CFD solutions obtained by VULCAN, a previously developed code used in engine analysis. The second experiment is a study of a supersonic combustor consisting of a diverging duct with single downstream-angled wall injector. Thus, the geometry is relatively simple and large regions of subsonic recirculating flow are avoided. The nominal entrance Mach number is 2 and the enthalpy of the test gas (hot air "simulant") is nominally that of Mach 7 flight. It was believed, on the basis of calculations performed that this would produce mixing-limited flow, that is to say, one for which chemical reaction to equilibrium proceeds at a much greater rate than mixing. It later proved that this was not the case. The primary experimental technique employed is coherent anti-Stokes Raman spectroscopy, known by its acronym CARS. The species probed is molecular nitrogen and the quantity measured is temperature. Intrusive probes, such as Pitot, total temperature, hot-wire, etc., are not used due to access difficulty and high heat flux in the combustor, and because they may alter the flow. CARS has several advantages over other optical methods. It is a relatively mature and well-understood technique. Signal levels are relatively high and the signal is in the form of a coherent (laser) beam that can be collected through small windows. Incoherent (non-CARS) interferences are rejected by spatial filtering.

Passive Ventricular Mechanics Modelling Using MRI of Structure and Function

PubMed Central

Wang, V.Y.; Lam, H.I.; Ennis, D.B.; Young, A.A.; Nash, M.P.

2009-01-01

Patients suffering from dilated cardiomyopathy or myocardial infarction can develop left ventricular (LV) diastolic impairment. The LV remodels its structure and function to adapt to pathophysiological changes in geometry and loading conditions and this remodeling process can alter the passive ventricular mechanics. In order to better understand passive ventricular mechanics, a LV finite element model was developed to incorporate physiological and mechanical information derived from in vivo magnetic resonance imaging (MRI) tissue tagging, in vivo LV cavity pressure recording and ex vivo diffusion tensor MRI (DTMRI) of a canine heart. MRI tissue tagging enables quantitative evaluation of cardiac mechanical function with high spatial and temporal resolution, whilst the direction of maximum water diffusion (the primary eigenvector) in each voxel of a DTMRI directly correlates with the myocardial fibre orientation. This model was customized to the geometry of the canine LV during diastasis by fitting the segmented epicardial and endocardial surface data from tagged MRI using nonlinear finite element fitting techniques. Myofibre orientations, extracted from DTMRI of the same heart, were incorporated into this geometric model using a free form deformation methodology. Pressure recordings, temporally synchronized to the tissue tagging MRI data, were used to simulate the LV deformation during diastole. Simulation of the diastolic LV mechanics allowed us to estimate the stiffness of the passive LV myocardium based on kinematic data obtained from tagged MRI. This integrated physiological model will allow more insight into the regional passive diastolic mechanics of the LV on an individualized basis, thereby improving our understanding of the underlying structural basis of mechanical dysfunction in pathological conditions. PMID:18982680

Imaging HIV-1 Tat Trafficking and Interactions by Engineered Green-Fluorescent-Protein Tagging

NASA Astrophysics Data System (ADS)

Beltram, Fabio

2002-03-01

The direct monitoring of protein function in live cells under physiologically relevant conditions is one of the most powerful and innovative methodologies for proteomics. Efficient florescent probes fully compatible with human-cell expression are the fundamental tools for these studies and their optimization opens the way to resolution at the single-protein level. Biological events involving protein pairs are also directly accessible thanks to tuning of protein-tag spectral properties and production of complementary pairs. Such pairs are characterized by overlapping absorption (for the acceptor tag) and emission (for the donor tag) spectra. By tagging the proteins of interest with acceptor and donor molecules, protein interaction can be directly visualized by FRET, fluorescent resonant energy transfer. In this talk we shall present the design by molecular dynamics calculations and the application of optimized green fluorescent proteins to the study of the human immunodeficiency virus HIV-1 proteomics. In particular trafficking and cellular interactions of HIV-1 transactivator protein Tat in live human cells will be presented. Tat localization and complex internalization pathways of exogenous molecules will be presented thanks to the peculiar optical properties of mutated GFPs. Cellular protein partners and subcellular interaction sites will be identified and directly visualized. The relevance of such results and of advanced spectroscopic and imaging techniques for a new level of understanding of biological processes and its significance for advancement in molecular biology will be underlined. A. Marcello et al., J. Biol. Chem. 276, 39220 (2001). R. Cinelli et al., Appl. Phys. Lett. 79, 3353 (2001).

Passive ventricular mechanics modelling using MRI of structure and function.

PubMed

Wang, V Y; Lam, H I; Ennis, D B; Young, A A; Nash, M P

2008-01-01

Patients suffering from dilated cardiomyopathy or myocardial infarction can develop left ventricular (LV) diastolic impairment. The LV remodels its structure and function to adapt to pathophysiological changes in geometry and loading conditions and this remodeling process can alter the passive ventricular mechanics. In order to better understand passive ventricular mechanics, a LV finite element model was developed to incorporate physiological and mechanical information derived from in vivo magnetic resonance imaging (MRI) tissue tagging, in vivo LV cavity pressure recording and ex vivo diffusion tensor MRI (DTMRI) of a canine heart. MRI tissue tagging enables quantitative evaluation of cardiac mechanical function with high spatial and temporal resolution, whilst the direction of maximum water diffusion (the primary eigenvector) in each voxel of a DTMRI directly correlates with the myocardial fibre orientation. This model was customized to the geometry of the canine LV during diastasis by fitting the segmented epicardial and endocardial surface data from tagged MRI using nonlinear finite element fitting techniques. Myofibre orientations, extracted from DTMRI of the same heart, were incorporated into this geometric model using a free form deformation methodology. Pressure recordings, temporally synchronized to the tissue tagging MRI data, were used to simulate the LV deformation during diastole. Simulation of the diastolic LV mechanics allowed us to estimate the stiffness of the passive LV myocardium based on kinematic data obtained from tagged MRI. This integrated physiological model will allow more insight into the regional passive diastolic mechanics of the LV on an individualized basis, thereby improving our understanding of the underlying structural basis of mechanical dysfunction in pathological conditions.

Multiple kernels learning-based biological entity relationship extraction method.

PubMed

Dongliang, Xu; Jingchang, Pan; Bailing, Wang

2017-09-20

Automatic extracting protein entity interaction information from biomedical literature can help to build protein relation network and design new drugs. There are more than 20 million literature abstracts included in MEDLINE, which is the most authoritative textual database in the field of biomedicine, and follow an exponential growth over time. This frantic expansion of the biomedical literature can often be difficult to absorb or manually analyze. Thus efficient and automated search engines are necessary to efficiently explore the biomedical literature using text mining techniques. The P, R, and F value of tag graph method in Aimed corpus are 50.82, 69.76, and 58.61%, respectively. The P, R, and F value of tag graph kernel method in other four evaluation corpuses are 2-5% higher than that of all-paths graph kernel. And The P, R and F value of feature kernel and tag graph kernel fuse methods is 53.43, 71.62 and 61.30%, respectively. The P, R and F value of feature kernel and tag graph kernel fuse methods is 55.47, 70.29 and 60.37%, respectively. It indicated that the performance of the two kinds of kernel fusion methods is better than that of simple kernel. In comparison with the all-paths graph kernel method, the tag graph kernel method is superior in terms of overall performance. Experiments show that the performance of the multi-kernels method is better than that of the three separate single-kernel method and the dual-mutually fused kernel method used hereof in five corpus sets.

Immunofluorescence Staining — EDRN Public Portal

Cancer.gov

Direct immunofluorescence method is used to detect the deposit of immunoglobulins, complement components, fibrinogen, etc. in tissues. This technique is usually performed on frozen sections. The primary antibody is conjugated to fluorescein binds directly with the antigen and can be detected by the fluorescent tag using a fluorescent microscope.

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

Hu, L; Yin, F; Cai, J

Purpose: To develop a methodology of constructing physiological-based virtual thorax phantom based on hyperpolarized (HP) gas tagging MRI for evaluating deformable image registration (DIR). Methods: Three healthy subjects were imaged at both the end-of-inhalation (EOI) and the end-of-exhalation (EOE) phases using a high-resolution (2.5mm isovoxel) 3D proton MRI, as well as a hybrid MRI which combines HP gas tagging MRI and a low-resolution (4.5mm isovoxel) proton MRI. A sparse tagging displacement vector field (tDVF) was derived from the HP gas tagging MRI by tracking the displacement of tagging grids between EOI and EOE. Using the tDVF and the high-resolution MRmore » images, we determined the motion model of the entire thorax in the following two steps: 1) the DVF inside of lungs was estimated based on the sparse tDVF using a novel multi-step natural neighbor interpolation method; 2) the DVF outside of lungs was estimated from the DIR between the EOI and EOE images (Velocity AI). The derived motion model was then applied to the high-resolution EOI image to create a deformed EOE image, forming the virtual phantom where the motion model provides the ground truth of deformation. Five DIR methods were evaluated using the developed virtual phantom. Errors in DVF magnitude (Em) and angle (Ea) were determined and compared for each DIR method. Results: Among the five DIR methods, free form deformation produced DVF results that are most closely resembling the ground truth (Em=1.04mm, Ea=6.63°). The two DIR methods based on B-spline produced comparable results (Em=2.04mm, Ea=13.66°; and Em =2.62mm, Ea=17.67°), and the two optical-flow methods produced least accurate results (Em=7.8mm; Ea=53.04°; Em=4.45mm, Ea=31.02°). Conclusion: A methodology for constructing physiological-based virtual thorax phantom based on HP gas tagging MRI has been developed. Initial evaluation demonstrated its potential as an effective tool for robust evaluation of DIR in the lung.« less

An efficient annotation and gene-expression derivation tool for Illumina Solexa datasets.

PubMed

Hosseini, Parsa; Tremblay, Arianne; Matthews, Benjamin F; Alkharouf, Nadim W

2010-07-02

The data produced by an Illumina flow cell with all eight lanes occupied, produces well over a terabyte worth of images with gigabytes of reads following sequence alignment. The ability to translate such reads into meaningful annotation is therefore of great concern and importance. Very easily, one can get flooded with such a great volume of textual, unannotated data irrespective of read quality or size. CASAVA, a optional analysis tool for Illumina sequencing experiments, enables the ability to understand INDEL detection, SNP information, and allele calling. To not only extract from such analysis, a measure of gene expression in the form of tag-counts, but furthermore to annotate such reads is therefore of significant value. We developed TASE (Tag counting and Analysis of Solexa Experiments), a rapid tag-counting and annotation software tool specifically designed for Illumina CASAVA sequencing datasets. Developed in Java and deployed using jTDS JDBC driver and a SQL Server backend, TASE provides an extremely fast means of calculating gene expression through tag-counts while annotating sequenced reads with the gene's presumed function, from any given CASAVA-build. Such a build is generated for both DNA and RNA sequencing. Analysis is broken into two distinct components: DNA sequence or read concatenation, followed by tag-counting and annotation. The end result produces output containing the homology-based functional annotation and respective gene expression measure signifying how many times sequenced reads were found within the genomic ranges of functional annotations. TASE is a powerful tool to facilitate the process of annotating a given Illumina Solexa sequencing dataset. Our results indicate that both homology-based annotation and tag-count analysis are achieved in very efficient times, providing researchers to delve deep in a given CASAVA-build and maximize information extraction from a sequencing dataset. TASE is specially designed to translate sequence data in a CASAVA-build into functional annotations while producing corresponding gene expression measurements. Achieving such analysis is executed in an ultrafast and highly efficient manner, whether the analysis be a single-read or paired-end sequencing experiment. TASE is a user-friendly and freely available application, allowing rapid analysis and annotation of any given Illumina Solexa sequencing dataset with ease.

Opening the black box: imaging nanoparticle transport with MRI

NASA Astrophysics Data System (ADS)

Phoenix, V.; Holmes, W. M.

2009-12-01

While most renown for its use in medicine, magnetic resonance imaging (MRI) has tremendous potential in the study of environmental processes. Its ability to non-invasively image inside materials that are opaque to other imaging methods (in particular light based techniques) is a particular strength. MRI has already been used, for example, to study fluid flow in rocks and image mass transport and biogeochemical processes in biofilms [1-4]. Here, we report of the use of MRI to image nanoparticle transport through porous geologic media (in this case packed gravel columns). Packed column experiments are key to understanding nanoparticulate transport in porous geologic media. Whilst highly informative, the data obtained can be a bulk average of a complex and heterogeneous array of interactions within the column. Natural environmental systems are often complex, displaying heterogeneity in geometry, hydrodynamics, geochemistry and microbiology throughout. MRI enables us to quantify better how this heterogeneity may influence nanoparticle transport and fate by enabling us to look inside the column and image the movement of nanoparticles within. To make the nanoparticle readily visible to MRI, it is labelled with a paramagnetic tag (commonly gadolinium). Indeed, a wide variety of off-the-shelf paramagnetically tagged nanoparticles and macromolecules are available, each with different properties enabling us to explore the impact of particle charge, size etc on their transport behaviour. In this preliminary study, packed columns of quartz or marble based gravels (approx 5 mm diameter) were first imaged to check their suitability for MR imaging. This was done as geologic material can contain sufficiently high concentrations of ferro- and paramagnetic ions to induce unwanted artefacts in the MR image. All gravels imaged (Rose quartz, Creswick quartz gravel and Ben Deulin white marble) produced minimal or no artefacts. A solution of the nanoparticle GadoCELLTrack (BioPAL), was prepared and connected to a 30 mm diameter flow cell containing rose quartz. GadoCELLTrack is a 30 nm diameter, neutrally charged gadolinium colloid. MR images were collected as the nanoparticle solution was pumped through the flow cell. These images were calibrated to provide fully quantitative maps of nanoparticle concentration at regular time intervals throughout the column. Such data can be used to help develop predictive models of nanoparticulate transport. [1] Holmes WM, Packer KJ (2003) Magnetic Resonance Imaging 21,389-391 [2] Seymour JD et al., (2004) Journal of Magnetic Resonance 167, 322-327 [3] McLean JS et al., (2008) ISME, 2, 121-131 [4] Phoenix et al., (2008) Applied and Environmental Microbiology, 74, 4934-4943

Estimation of mussel population response to hydrologic alteration in a southeastern U.S. stream

USGS Publications Warehouse

Peterson, J.T.; Wisniewski, J.M.; Shea, C.P.; Rhett, Jackson C.

2011-01-01

The southeastern United States has experienced severe, recurrent drought, rapid human population growth, and increasing agricultural irrigation during recent decades, resulting in greater demand for the water resources. During the same time period, freshwater mussels (Unioniformes) in the region have experienced substantial population declines. Consequently, there is growing interest in determining how mussel population declines are related to activities associated with water resource development. Determining the causes of mussel population declines requires, in part, an understanding of the factors influencing mussel population dynamics. We developed Pradel reverse-time, tag-recapture models to estimate survival, recruitment, and population growth rates for three federally endangered mussel species in the Apalachicola- Chattahoochee-Flint River Basin, Georgia. The models were parameterized using mussel tag-recapture data collected over five consecutive years from Sawhatchee Creek, located in southwestern Georgia. Model estimates indicated that mussel survival was strongly and negatively related to high flows during the summer, whereas recruitment was strongly and positively related to flows during the spring and summer. Using these models, we simulated mussel population dynamics under historic (1940-1969) and current (1980-2008) flow regimes and under increasing levels of water use to evaluate the relative effectiveness of alternative minimum flow regulations. The simulations indicated that the probability of simulated mussel population extinction was at least 8 times greater under current hydrologic regimes. In addition, simulations of mussel extinction under varying levels of water use indicated that the relative risk of extinction increased with increased water use across a range of minimum flow regulations. The simulation results also indicated that our estimates of the effects of water use on mussel extinction were influenced by the assumptions about the dynamics of the system, highlighting the need for further study of mussel population dynamics. ?? 2011 Springer Science+Business Media, LLC (outside the USA).

Characterization of microbial associations with methanotrophic archaea and sulfate-reducing bacteria through statistical comparison of nested Magneto-FISH enrichments

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

Trembath-Reichert, Elizabeth; Case, David H.; Orphan, Victoria J.

Methane seep systems along continental margins host diverse and dynamic microbial assemblages, sustained in large part through the microbially mediated process of sulfate-coupled Anaerobic Oxidation of Methane (AOM). This methanotrophic metabolism has been linked to consortia of anaerobic methane-oxidizing archaea (ANME) and sulfate-reducing bacteria (SRB). These two groups are the focus of numerous studies; however, less is known about the wide diversity of other seep associated microorganisms. We selected a hierarchical set of FISH probes targeting a range ofDeltaproteobacteriadiversity. Using the Magneto-FISH enrichment technique, we then magnetically captured CARD-FISH hybridized cells and their physically associated microorganisms from a methane seepmore » sediment incubation. DNA from nested Magneto-FISH experiments was analyzed using Illumina tag 16S rRNA gene sequencing (iTag). Enrichment success and potential bias with iTag was evaluated in the context of full-length 16S rRNA gene clone libraries, CARD-FISH, functional gene clone libraries, and iTag mock communities. We determined commonly used Earth Microbiome Project (EMP) iTAG primers introduced bias in some common methane seep microbial taxa that reduced the ability to directly compare OTU relative abundances within a sample, but comparison of relative abundances between samples (in nearly all cases) and whole community-based analyses were robust. The iTag dataset was subjected to statistical co-occurrence measures of the most abundant OTUs to determine which taxa in this dataset were most correlated across all samples. In addition, many non-canonical microbial partnerships were statistically significant in our co-occurrence network analysis, most of which were not recovered with conventional clone library sequencing, demonstrating the utility of combining Magneto-FISH and iTag sequencing methods for hypothesis generation of associations within complex microbial communities. Network analysis pointed to many co-occurrences containing putatively heterotrophic, candidate phyla such as OD1, Atribacteria, MBG-B, and Hyd24-12 and the potential for complex sulfur cycling involving Epsilon-, Delta-, and Gammaproteobacteria in methane seep ecosystems.« less

Characterization of microbial associations with methanotrophic archaea and sulfate-reducing bacteria through statistical comparison of nested Magneto-FISH enrichments

DOE PAGES

Trembath-Reichert, Elizabeth; Case, David H.; Orphan, Victoria J.

2016-04-18

Methane seep systems along continental margins host diverse and dynamic microbial assemblages, sustained in large part through the microbially mediated process of sulfate-coupled Anaerobic Oxidation of Methane (AOM). This methanotrophic metabolism has been linked to consortia of anaerobic methane-oxidizing archaea (ANME) and sulfate-reducing bacteria (SRB). These two groups are the focus of numerous studies; however, less is known about the wide diversity of other seep associated microorganisms. We selected a hierarchical set of FISH probes targeting a range ofDeltaproteobacteriadiversity. Using the Magneto-FISH enrichment technique, we then magnetically captured CARD-FISH hybridized cells and their physically associated microorganisms from a methane seepmore » sediment incubation. DNA from nested Magneto-FISH experiments was analyzed using Illumina tag 16S rRNA gene sequencing (iTag). Enrichment success and potential bias with iTag was evaluated in the context of full-length 16S rRNA gene clone libraries, CARD-FISH, functional gene clone libraries, and iTag mock communities. We determined commonly used Earth Microbiome Project (EMP) iTAG primers introduced bias in some common methane seep microbial taxa that reduced the ability to directly compare OTU relative abundances within a sample, but comparison of relative abundances between samples (in nearly all cases) and whole community-based analyses were robust. The iTag dataset was subjected to statistical co-occurrence measures of the most abundant OTUs to determine which taxa in this dataset were most correlated across all samples. In addition, many non-canonical microbial partnerships were statistically significant in our co-occurrence network analysis, most of which were not recovered with conventional clone library sequencing, demonstrating the utility of combining Magneto-FISH and iTag sequencing methods for hypothesis generation of associations within complex microbial communities. Network analysis pointed to many co-occurrences containing putatively heterotrophic, candidate phyla such as OD1, Atribacteria, MBG-B, and Hyd24-12 and the potential for complex sulfur cycling involving Epsilon-, Delta-, and Gammaproteobacteria in methane seep ecosystems.« less

Characterization of microbial associations with methanotrophic archaea and sulfate-reducing bacteria through statistical comparison of nested Magneto-FISH enrichments.

PubMed

Trembath-Reichert, Elizabeth; Case, David H; Orphan, Victoria J

2016-01-01

Methane seep systems along continental margins host diverse and dynamic microbial assemblages, sustained in large part through the microbially mediated process of sulfate-coupled Anaerobic Oxidation of Methane (AOM). This methanotrophic metabolism has been linked to consortia of anaerobic methane-oxidizing archaea (ANME) and sulfate-reducing bacteria (SRB). These two groups are the focus of numerous studies; however, less is known about the wide diversity of other seep associated microorganisms. We selected a hierarchical set of FISH probes targeting a range of Deltaproteobacteria diversity. Using the Magneto-FISH enrichment technique, we then magnetically captured CARD-FISH hybridized cells and their physically associated microorganisms from a methane seep sediment incubation. DNA from nested Magneto-FISH experiments was analyzed using Illumina tag 16S rRNA gene sequencing (iTag). Enrichment success and potential bias with iTag was evaluated in the context of full-length 16S rRNA gene clone libraries, CARD-FISH, functional gene clone libraries, and iTag mock communities. We determined commonly used Earth Microbiome Project (EMP) iTAG primers introduced bias in some common methane seep microbial taxa that reduced the ability to directly compare OTU relative abundances within a sample, but comparison of relative abundances between samples (in nearly all cases) and whole community-based analyses were robust. The iTag dataset was subjected to statistical co-occurrence measures of the most abundant OTUs to determine which taxa in this dataset were most correlated across all samples. Many non-canonical microbial partnerships were statistically significant in our co-occurrence network analysis, most of which were not recovered with conventional clone library sequencing, demonstrating the utility of combining Magneto-FISH and iTag sequencing methods for hypothesis generation of associations within complex microbial communities. Network analysis pointed to many co-occurrences containing putatively heterotrophic, candidate phyla such as OD1, Atribacteria, MBG-B, and Hyd24-12 and the potential for complex sulfur cycling involving Epsilon-, Delta-, and Gammaproteobacteria in methane seep ecosystems.

Extending the versatility of the Hemetsberger-Knittel indole synthesis through microwave and flow chemistry.

PubMed

Ranasinghe, Nadeesha; Jones, Graham B

2013-03-15

Microwave, flow and combination methodologies have been applied to the synthesis of a number of substituted indoles. Based on the Hemetsberger-Knittel (HK) process, modifications allow formation of products rapidly and in high yield. Adapting the methodology allows formation of 2-unsubstituted indoles and derivatives, and a route to analogs of the antitumor agent PLX-4032 is demonstrated. The utility of the HK substrates is further demonstrated through bioconjugation and subsequent ring closure and via Huisgen type [3+2] cycloaddition chemistry, allowing formation of peptide adducts which can be subsequently labeled with fluorine tags. Copyright © 2013 Elsevier Ltd. All rights reserved.

Oh! Web 2.0, Virtual Reference Service 2.0, Tools & Techniques (II)

ERIC Educational Resources Information Center

Arya, Harsh Bardhan; Mishra, J. K.

2012-01-01

The paper describes the theory and definition of the practice of librarianship, specifically addressing how Web 2.0 technologies (tools) such as synchronous messaging, collaborative reference service and streaming media, blogs, wikis, social networks, social bookmarking tools, tagging, RSS feeds, and mashups might intimate changes and how…

An Art of Resistance: From the Street to the Classroom

ERIC Educational Resources Information Center

Chung, Sheng Kuan

2009-01-01

Rooted in graffiti culture and its attitude toward the world, street art is regarded as a postgraffiti movement. Street art encompasses a wide array of media and techniques, such as traditional spray-painted tags, stickers, stencils, posters, photocopies, murals, paper cutouts, mosaics, street installations, performances, and video projections…

Biochemist's Toolbox

ERIC Educational Resources Information Center

Bakhtiar, Ray

2013-01-01

Surface plasmon resonance (SPR) spectroscopy is a powerful, label-free technique to monitor noncovalent molecular interactions in real time and in a noninvasive fashion. As a label-free assay, SPR does not require tags, dyes, or specialized reagents (e.g., enzymes-substrate complexes) to elicit a visible or a fluorescence signal. During the last…

I-DIRT, a general method for distinguishing between specific and nonspecific protein interactions.

PubMed

Tackett, Alan J; DeGrasse, Jeffrey A; Sekedat, Matthew D; Oeffinger, Marlene; Rout, Michael P; Chait, Brian T

2005-01-01

Isolation of protein complexes via affinity-tagged proteins provides a powerful tool for studying biological systems, but the technique is often compromised by co-enrichment of nonspecifically interacting proteins. We describe a new technique (I-DIRT) that distinguishes contaminants from bona fide interactors in immunopurifications, overcoming this most challenging problem in defining protein complexes. I-DIRT will be of broad value for studying protein complexes in biological systems that can be metabolically labeled.

Molecular Tagging Velocimetry Development for In-situ Measurement in High-Temperature Test Facility

NASA Technical Reports Server (NTRS)

Andre, Matthieu A.; Bardet, Philippe M.; Burns, Ross A.; Danehy, Paul M.

2015-01-01

The High Temperature Test Facility, HTTF, at Oregon State University (OSU) is an integral-effect test facility designed to model the behavior of a Very High Temperature Gas Reactor (VHTR) during a Depressurized Conduction Cooldown (DCC) event. It also has the ability to conduct limited investigations into the progression of a Pressurized Conduction Cooldown (PCC) event in addition to phenomena occurring during normal operations. Both of these phenomena will be studied with in-situ velocity field measurements. Experimental measurements of velocity are critical to provide proper boundary conditions to validate CFD codes, as well as developing correlations for system level codes, such as RELAP5 (http://www4vip.inl.gov/relap5/). Such data will be the first acquired in the HTTF and will introduce a diagnostic with numerous other applications to the field of nuclear thermal hydraulics. A laser-based optical diagnostic under development at The George Washington University (GWU) is presented; the technique is demonstrated with velocity data obtained in ambient temperature air, and adaptation to high-pressure, high-temperature flow is discussed.

An optimized velocity selective arterial spin labeling module with reduced eddy current sensitivity for improved perfusion quantification.

PubMed

Meakin, James A; Jezzard, Peter

2013-03-01

Velocity-selective (VS) arterial spin labeling is a promising method for measuring perfusion in areas of slow or collateral flow by eliminating the bolus arrival delay associated with other spin labeling techniques. However, B(0) and B(1) inhomogeneities and eddy currents during the VS preparation hinder accurate quantification of perfusion with VS arterial spin labeling. In this study, it is demonstrated through simulations and experiments in healthy volunteers that eddy currents cause erroneous tagging of static tissue. Consequently, mean gray matter perfusion is overestimated by up to a factor of 2, depending on the VS preparation used. A novel eight-segment B(1) insensitive rotation VS preparation is proposed to reduce eddy current effects while maintaining the B(0) and B(1) insensitivity of previous preparations. Compared to two previous VS preparations, the eight-segment B(1) insensitive rotation is the most robust to eddy currents and should improve the quality and reliability of VS arterial spin labeling measurements in future studies. Copyright © 2012 Wiley Periodicals, Inc.

Smolt Monitoring at the Head of Lower Granite Reservoir and Lower Granite Dam; Smolt Monitoring by Federal and Non-Federal Entities, 2001-2002 Annual Report.

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

Buettner, Edwin W.; Putnam, Scott A.

This project monitored the daily passage of chinook salmon Oncorhynchus tshawytscha, steelhead trout O. mykiss, and sockeye salmon smolts O. nerka during the 2001 spring out-migration at migrant traps on the Snake River and Salmon River. In 2001 fish management agencies released significant numbers of hatchery chinook salmon and steelhead trout above Lower Granite Dam that were not marked with a fin clip or coded-wire tag. Generally, these fish were distinguishable from wild fish by the occurrence of fin erosion. Total annual hatchery chinook salmon catch at the Snake River trap was 11% of the 2000 numbers. The wild chinookmore » catch was 3% of the previous year's catch. Hatchery steelhead trout catch was 49% of 2000 numbers. Wild steelhead trout catch was 69% of 2000 numbers. The Snake River trap collected 28 age-0 chinook salmon. During 2001 the Snake River trap captured zero hatchery and zero wild/natural sockeye salmon and six hatchery coho salmon O. kisutch. Differences in trap catch between years are due to fluctuations not only in smolt production, but also differences in trap efficiency and duration of trap operation associated with flow. The significant reduction in catch during 2001 was due to a reduction in hatchery chinook production (60% of 2000 release) and due to extreme low flows. Trap operations began on March 11 and were terminated on June 29. The trap was out of operation for a total of two days due to mechanical failure or debris. Hatchery chinook salmon catch at the Salmon River trap was 47% and wild chinook salmon catch was 67% of 2000 numbers. The hatchery steelhead trout collection in 2001 was 178% of the 2000 numbers. Wild steelhead trout collection in 2001 was 145% of the previous year's catch. Trap operations began on March 11 and were terminated on June 8 due to the end of the smolt monitoring season. There were no days where the trap was out of operation due to high flow or debris. The decrease in hatchery chinook catch in 2001 was due to a reduction in hatchery production (39% of 2000 releases). The increase in hatchery and wild steelhead trap catch is due to the ability to operate the trap in the thalweg for a longer period of time because of the extreme low flow condition in 2001. Travel time (d) and migration rate (km/d) through Lower Granite Reservoir for PIT-tagged chinook salmon and steelhead trout marked at the head of the reservoir were affected by discharge. There were not enough hatchery and wild chinook salmon tagged at the Snake River trap in 2001 to allow migration rate/discharge analysis. For steelhead trout tagged at the Snake River trap, statistical analysis of 2001 data detected a significant relation between migration rate and Lower Granite Reservoir inflow discharge. For hatchery and wild steelhead trout, there was a 2.2-fold and a 1.5-fold increase in migration rate in, respectively, between 50 and 100 kcfs. Travel time and migration rate to Lower Granite Dam for fish marked at the Salmon River trap were calculated. Statistical analysis of the 2001 data detected a significant relation between migration rate and Lower Granite Reservoir inflow discharge for hatchery and wild chinook salmon and hatchery and wild steelhead trout. Migration rate increased 3.7-fold for hatchery chinook salmon and 2.5-fold for wild chinook salmon between 50 and 100 kcfs. For hatchery steelhead there was a 1.6-fold increase in migration rate, and for wild steelhead trout there was a 2.2-fold increase between 50 kcfs and 100 kcfs. Fish tagged with passive integrated transponder (PIT) tags at the Snake River trap were interrogated at four dams with PIT tag detection systems (Lower Granite, Little Goose, Lower Monumental, and McNary dams). Because of the addition of the fourth interrogation site (Lower Monumental) in 1993, cumulative interrogation data is not comparable with the prior five years (1988-1992). Cumulative interrogations at the four dams for fish marked at the Snake River trap were 86% for hatchery chinook, 70% for wild chinook, 71% for hatchery steelhead, and 89% for wild steelhead. Cumulative interrogations at the four dams for fish marked at the Salmon River trap were 74% for hatchery chinook, 83% for wild chinook salmon, 75% for hatchery steelhead trout, and 81% for wild steelhead trout.« less

Long range ultra-high frequency (UHF) radio frequency identification (RFID) antenna design

NASA Astrophysics Data System (ADS)

Reynolds, Nathan D.

There is an ever-increasing demand for radio frequency identification (RFID) tags that are passive, long range, and mountable on multiple surfaces. Currently, RFID technology is utilized in numerous applications such as supply chain management, access control, and public transportation. With the combination of sensory systems in recent years, the applications of RFID technology have been extended beyond tracking and identifying. This extension includes applications such as environmental monitoring and healthcare applications. The available sensory systems usually operate in the medium or high frequency bands and have a low read range. However, the range limitations of these systems are being overcome by the development of RFID sensors focused on utilizing tags in the ultra-high frequency (UHF) band. Generally, RFID tags have to be mounted to the object that is being identified. Often the objects requiring identification are metallic. The inherent properties of metallic objects have substantial effects on nearby electromagnetic radiation; therefore, the operation of the tag antenna is affected when mounted on a metallic surface. This outlines one of the most challenging problems for RFID systems today: the optimization of tag antenna performance in a complex environment. In this research, a novel UHF RFID tag antenna, which has a low profile, long range, and is mountable on metallic surfaces, is designed analytically and simulated using a 3-D electromagnetic simulator, ANSYS HFSS. A microstrip patch antenna is selected as the antenna structure, as patch antennas are low profile and suitable for mounting on metallic surfaces. Matching and theoretical models of the microstrip patch antenna are investigated. Once matching and theory of a microstrip patch antenna is thoroughly understood, a unique design technique using electromagnetic band gap (EBG) structures is explored. This research shows that the utilization of an EBG structure in the patch antenna design yields an improvement in gain, or range, and in the ability to be mounted on multiple metallic surfaces.

Air Ground Integration and the Brigade Combat Team

DTIC Science & Technology

2013-06-13

Theater Air Control System TADIL-J Tactical Digital Information Link-J TAGS Theater Air Ground System TAIS Tactical Air Integration System TBMCS Theater...during planning and execution. This system interacts with the Theater Battle Management Core System ( TBMCS ) used by the JAOC to build and disseminate...control nodes within the AAGS, in conjunction with the interoperability with the TBMCS and Army mission command systems facilitates information flow during

Genetic studies of Northern Goshawks (Accipiter gentilis): genetic tagging and individual identification from feathers, and determining phylogeography, gene flow and population history for Goshawks in North America

Treesearch

Shelley Bayard de Volo

2008-01-01

The Northern Goshawk (Accipiter gentilis) is a large, highly mobile, mostly nonmigratory and widespread forest raptor. It ranges across the Boreal forests from Alaska to Newfoundland, and south into forests of the Great Lakes, and montane regions of the Appalachian, Cascade, Sierra Nevada, Rocky Mountain, and Sierra Madre Occidental.

High-Performance Java Codes for Computational Fluid Dynamics

NASA Technical Reports Server (NTRS)

Riley, Christopher; Chatterjee, Siddhartha; Biswas, Rupak; Biegel, Bryan (Technical Monitor)

2001-01-01

The computational science community is reluctant to write large-scale computationally -intensive applications in Java due to concerns over Java's poor performance, despite the claimed software engineering advantages of its object-oriented features. Naive Java implementations of numerical algorithms can perform poorly compared to corresponding Fortran or C implementations. To achieve high performance, Java applications must be designed with good performance as a primary goal. This paper presents the object-oriented design and implementation of two real-world applications from the field of Computational Fluid Dynamics (CFD): a finite-volume fluid flow solver (LAURA, from NASA Langley Research Center), and an unstructured mesh adaptation algorithm (2D_TAG, from NASA Ames Research Center). This work builds on our previous experience with the design of high-performance numerical libraries in Java. We examine the performance of the applications using the currently available Java infrastructure and show that the Java version of the flow solver LAURA performs almost within a factor of 2 of the original procedural version. Our Java version of the mesh adaptation algorithm 2D_TAG performs within a factor of 1.5 of its original procedural version on certain platforms. Our results demonstrate that object-oriented software design principles are not necessarily inimical to high performance.

Plastic debris retention and exportation by a mangrove forest patch.

PubMed

Ivar do Sul, Juliana A; Costa, Monica F; Silva-Cavalcanti, Jacqueline S; Araújo, Maria Christina B

2014-01-15

An experiment observed the behavior of selected tagged plastic items deliberately released in different habitats of a tropical mangrove forest in NE Brazil in late rainy (September) and late dry (March) seasons. Significant differences were not reported among seasons. However, marine debris retention varied among habitats, according to characteristics such as hydrodynamic (i.e., flow rates and volume transported) and relative vegetation (Rhizophora mangle) height and density. The highest grounds retained significantly more items when compared to the borders of the river and the tidal creek. Among the used tagged items, PET bottles were more observed and margarine tubs were less observed, being easily transported to adjacent habitats. Plastic bags were the items most retained near the releasing site. The balance between items retained and items lost was positive, demonstrating that mangrove forests tend to retain plastic marine debris for long periods (months-years). Copyright © 2013 Elsevier Ltd. All rights reserved.

Molecular imaging and sensing using plasmonic nanoparticles

NASA Astrophysics Data System (ADS)

Crow, Matthew James

Noble metal nanoparticles exhibit unique optical properties that are beneficial to a variety of applications, including molecular imaging. The large scattering cross sections of nanoparticles provide high contrast necessary for biomarkers. Unlike alternative contrast agents, nanoparticles provide refractive index sensitivity revealing information regarding the local cellular environment. Altering the shape and composition of the nanoparticle shifts the peak resonant wavelength of scattered light, allowing for implementation of multiple spectrally distinct tags. In this project, nanoparticles that scatter in different spectral windows are functionalized with various antibodies recognizing extra-cellular receptors integral to cancer progression. A hyperspectral imaging system is developed, allowing for visualization and spectral characterization of cells labeled with these conjugates. Various molecular imaging and microspectroscopy applications of plasmonic nanoparticles are then investigated. First, anti-EGFR gold nanospheres are shown to quantitatively measure receptor expression with similar performance to fluorescence assays. Second, anti-EGFR gold nanorods and novel anti-IGF-1R silver nanospheres are implemented to indicate local cellular refractive indices. Third, because biosensing capabilities of nanoparticle tags may be limited by plasmonic coupling, polarization mapping is investigated as a method to discern these effects. Fourth, plasmonic coupling is tested to monitor HER-2 dimerization. Experiments reveal the interparticle conformation of proximal HER-2 bound labels, required for plasmonic coupling-enhanced dielectric sensing. Fifth, all three functionalized plasmonic tags are implemented simultaneously to indicate clinically relevant cell immunophenotype information and changes in the cellular dielectric environment. Finally, flow cytometry experiments are conducted utilizing the anti-EGFR nanorod tag to demonstrate profiling of receptor expression distribution and potential increased multiplexing capability.

Interaction with a kinesin-2 tail propels choline acetyltransferase flow towards synapse

PubMed Central

Sadananda, Aparna; Hamid, Runa; Doodhi, Harinath; Ghosal, Debnath; Girotra, Mukul; Jana, Swadhin Chandra; Ray, Krishanu

2012-01-01

Bulk flow constitutes a substantial part of the slow transport of soluble proteins in axons. Though the underlying mechanism is unclear, evidences indicate that intermittent, kinesin based movement of large protein-aggregates aids this process. Choline acetyl-transferase (ChAT), a soluble enzyme catalyzing acetylcholine synthesis, propagates towards synapse at an intermediate, slow rate. The presynaptic enrichment of ChAT requires heterotrimeric kinesin-2, comprising KLP64D, KLP68D and DmKAP, in Drosophila. Here, we show that the bulk flow of a recombinant Green Fluorescent Protein-tagged ChAT (GFP::ChAT), in Drosophila axons, lacks particulate features. It occurs for a brief period during the larval stages. In addition, both the endogenous ChAT and GFP::ChAT directly bind to the KLP64D tail, which is essential for the GFP::ChAT entry and anterograde flow in axon. These evidences suggest that a direct interaction with motor proteins could regulate the bulk flow of soluble proteins, and thus establish their asymmetric distribution. PMID:22486887

Sediment routing through channel confluences: RFID tracer experiments from a gravel-bed river headwaters

NASA Astrophysics Data System (ADS)

Imhoff, K.; Wilcox, A. C.

2014-12-01

Tributary confluences may significantly impact large-scale patterns of sediment transport because of their role in connecting individual streams in a network. These unique locations feature complex flow structures and geomorphic features, and may represent ecological hotspots. Sediment transport across confluences is poorly understood, however. We present research on coarse sediment transport and dispersion through confluences using sediment tracers in the East Fork Bitterroot River, Montana, USA. We tagged a range of gravel (>40 mm) and cobble particles with Radio Frequency Identification (RFID) tags and painted smaller (10-40 mm) gravels, and then we traced them through confluences in a montane river's headwaters. We measured the effects of confluences on dispersion, path length, and depositional location and compare properties of sediment routing with a non-confluence control reach. We also measured topographic change through repeat bed surveys and combined topography, hydraulics, and tracer measurements to calculate basal shear and critical Shields stresses for different grain sizes. Field observations suggest that tagged particles in confluences routed along flanks of scour holes in confluences, with sediment depositing further downstream along bank-lateral bars than within the channel thalweg. Travel distances of RFID-tagged particles ranged up to 35 meters from original seeding points, with initial recovery rates of RFID-tagged tracers ranging between 84-89%. In both confluence and control reaches only partial mobility was observed within the entire tracer population, suggesting a hiding effect imposed by the roughness of the bed. Particles seeded in the channel thalweg experienced further travel distances than those seeded towards the banks and on bars. Differences in dispersion between confluence and control reaches are implied by field observation. This study quantified patterns of sediment routing within confluences and provided insight to the importance of confluences in large-scale sediment transport through gravel-bed rivers.

Developmental staging of male murine embryonic gonad by SAGE analysis

PubMed Central

Lee, Tin-Lap; Li, Yunmin; Alba, Diana; Vong, Queenie P.; Wu, Shao-Ming; Baxendale, Vanessa; Rennert, Owen M.; Lau, Yun-Fai Chris; Chan, Wai-Yee

2012-01-01

Despite the identification of key genes such as Sry integral to embryonic gonadal development, the genomic classification and identification of chromosomal activation of this process is still poorly understood. To better understand the genetic regulation of gonadal development, we performed Serial Analysis of Gene Expression (SAGE) to profile the genes and novel transcripts, and an average of 152,000 tags from male embryonic gonads at E10.5 (embryonic day 10.5), E11.5, E12.5, E13.5, E15.5 and E17.5 were analyzed. A total of 275,583 non-singleton tags that do not map to any annotated sequence were identified in the six gonad libraries, and 47,255 tags were mapped to 24,975 annotated sequences, among which 987 sequences were uncharacterized. Utilizing an unsupervised pattern identification technique, we established molecular staging of male gonadal development. Rather than providing a static descriptive analysis, we developed algorithms to cluster the SAGE data and assign SAGE tags to a corresponding chromosomal position; these data are displayed in chromosome graphic format. A prominent increase in global genomic activity from E10.5 to E17.5 was observed. Important chromosomal regions related to the developmental processes were identified and validated based on established mouse models with developmental disorders. These regions may represent markers for early diagnosis for disorders of male gonad development as well as potential treatment targets. PMID:19376482

A Novel RFID-Based Sensing Method for Low-Cost Bolt Loosening Monitoring

PubMed Central

Wu, Jian; Cui, Xingmei; Xu, Yunpeng

2016-01-01

In coal mines, bolt loosening in the cage guide is affected by the harsh environmental factors and cage hoist vibration, leading to significant threats to work safety. It is crucial, to this effect, to successfully detect the status of multipoint bolts of guide structures. This paper proposes a system to monitor bolt status in harsh environments established based on the RFID technique. A proof-of-concept model was demonstrated consisting of a bolt gearing system, passive UHF RFID tags, a reader, and monitoring software. A tinfoil metal film is fixed on the retaining plate and an RFID tag bonded to a large gear, with the bolt to be detected fixed in the center of a smaller gear. The radio-frequency signal cannot be received by the reader if the tag is completely obscured by the tinfoil, and if the bolt is loose, the tag’s antenna is exposed when the gear revolves. A radio-frequency signal that carries corresponding bolt’s information is transmitted by the RFID tag to the RFID reader due to coil coupling, identifying loose bolt location and reporting them in the software. Confirmatory test results revealed that the system indeed successfully detects bolt loosening and comparative test results (based on a reed switch multipoint bolt loosening monitor system) provided valuable information regarding the strengths and weaknesses of the proposed system. PMID:26828498

The behaviour and recovery of juvenile lemon sharks Negaprion brevirostris in response to external accelerometer tag attachment.

PubMed

Bullock, R W; Guttridge, T L; Cowx, I G; Elliott, M; Gruber, S H

2015-12-01

Behavioural responses of lemon sharks Negaprion brevirostris to a fin-mounted tag package (CEFAS G6A tri-axial accelerometer with epoxied Sonotronics PT4 acoustic transmitter) were measured in a controlled captive environment (n = 10, total length, LT range 80-140 cm) and in free-ranging sharks upon release (n = 7, LT range 100-160 cm). No changes were detected in behaviour (i.e. swimming speed, tailbeat frequency, time spent resting and frequency of chafing) between control and tagged captive shark trials, suggesting that the tag package itself does not alter behaviour. In the free-ranging trials, an initial period of elevated swimming activity was found in all individuals (represented by overall dynamic body acceleration). Negaprion brevirostris, however, appeared to recover quickly, returning to a steady swimming state between 2 and 35 min after release. Post-release tracking found that all sharks swim immediately for the shoreline and remain within 100 m of shore for prolonged periods. Hence, although N. brevirostris are capable of quick adaptation to stressors and demonstrate rapid recovery in terms of activity, tracking data suggest that they may modify their spatial use patterns post release. This research is important in separating deviation in behaviour due to environmental stressors from artefacts caused by experimental techniques. © 2015 The Fisheries Society of the British Isles.

Building and using a statistical 3D motion atlas for analyzing myocardial contraction in MRI

NASA Astrophysics Data System (ADS)

Rougon, Nicolas F.; Petitjean, Caroline; Preteux, Francoise J.

2004-05-01

We address the issue of modeling and quantifying myocardial contraction from 4D MR sequences, and present an unsupervised approach for building and using a statistical 3D motion atlas for the normal heart. This approach relies on a state-of-the-art variational non rigid registration (NRR) technique using generalized information measures, which allows for robust intra-subject motion estimation and inter-subject anatomical alignment. The atlas is built from a collection of jointly acquired tagged and cine MR exams in short- and long-axis views. Subject-specific non parametric motion estimates are first obtained by incremental NRR of tagged images onto the end-diastolic (ED) frame. Individual motion data are then transformed into the coordinate system of a reference subject using subject-to-reference mappings derived by NRR of cine ED images. Finally, principal component analysis of aligned motion data is performed for each cardiac phase, yielding a mean model and a set of eigenfields encoding kinematic ariability. The latter define an organ-dedicated hierarchical motion basis which enables parametric motion measurement from arbitrary tagged MR exams. To this end, the atlas is transformed into subject coordinates by reference-to-subject NRR of ED cine frames. Atlas-based motion estimation is then achieved by parametric NRR of tagged images onto the ED frame, yielding a compact description of myocardial contraction during diastole.

Optical Lock-In Detection of FRET Using Synthetic and Genetically Encoded Optical Switches

PubMed Central

Mao, Shu; Benninger, Richard K. P.; Yan, Yuling; Petchprayoon, Chutima; Jackson, David; Easley, Christopher J.; Piston, David W.; Marriott, Gerard

2008-01-01

The Förster resonance energy transfer (FRET) technique is widely used for studying protein interactions within live cells. The effectiveness and sensitivity of determining FRET, however, can be reduced by photobleaching, cross talk, autofluorescence, and unlabeled, endogenous proteins. We present a FRET imaging method using an optical switch probe, Nitrobenzospiropyran (NitroBIPS), which substantially improves the sensitivity of detection to <1% FRET efficiency. Through orthogonal optical control of the colorful merocyanine and colorless spiro states of the NitroBIPS acceptor, donor fluorescence can be measured both in the absence and presence of FRET in the same FRET pair in the same cell. A SNAP-tag approach is used to generate a green fluorescent protein-alkylguaninetransferase fusion protein (GFP-AGT) that is labeled with benzylguanine-NitroBIPS. In vivo imaging studies on this green fluorescent protein-alkylguaninetransferase (GFP-AGT) (NitroBIPS) complex, employing optical lock-in detection of FRET, allow unambiguous resolution of FRET efficiencies below 1%, equivalent to a few percent of donor-tagged proteins in complexes with acceptor-tagged proteins. PMID:18281383

Covalent immobilization of lipase onto aminopropyl-functionalized hydroxyapatite-encapsulated-γ-Fe2O3 nanoparticles: A magnetic biocatalyst for interesterification of soybean oil.

PubMed

Xie, Wenlei; Zang, Xuezhen

2017-07-15

Hydroxyapatite-encapsulated γ-Fe 2 O 3 nanoparticles were prepared, and lipase from Candida rugosa was then covalently bound onto the magnetic materials via covalent linkages. The magnetic carrier and immobilized lipase were characterized by enzyme activity assays, XRD, FT-IR, TEM, VSM and N 2 adsorption-desorption techniques. Results demonstrated that γ-Fe 2 O 3 nanoparticles were coated with the hydroxyapatite, and the lipase was indeed tethered to the magnetic carriers without damage to their structure. The immobilized lipase showed a strong magnetic responsiveness and displayed high catalytic activities towards the interesterification of soybean oil. The interesterified products were evaluated for their total fatty acid (FA) composition, slip melting point (SMP), iodine value, triacylglycerols (TAGs) profile and FA composition at sn-2 position in TAGs. The FA positional distributions and TAG species significantly changed after the enzymatic interesterification. Besides this, the interesterified products showed an obvious reduction in their SMP in comparison with the physical blends. Copyright © 2017 Elsevier Ltd. All rights reserved.

Precise timing correlation in telemetry recording and processing systems

NASA Technical Reports Server (NTRS)

Pickett, R. B.; Matthews, F. L.

1973-01-01

Independent PCM telemetry data signals received from missiles must be correlated to within + or - 100 microseconds for comparison with radar data. Tests have been conducted to determine RF antenna receiving system delays; delays associated with wideband analog tape recorders used in the recording, dubbing and repdocuing processes; and uncertainties associated with computer processed time tag data. Several methods used in the recording of timing are evaluated. Through the application of a special time tagging technique, the cumulative timing bias from all sources is determined and the bias removed from final data. Conclusions show that relative time differences in receiving, recording, playback and processing of two telemetry links can be accomplished with a + or - 4 microseconds accuracy. In addition, the absolute time tag error (with respect to UTC) can be reduced to less than 15 microseconds. This investigation is believed to be the first attempt to identify the individual error contributions within the telemetry system and to describe the methods of error reduction within the telemetry system and to describe the methods of error reduction and correction.

Tagging and purifying proteins to teach molecular biology and advanced biochemistry.

PubMed

Roecklein-Canfield, Jennifer A; Lopilato, Jane

2004-11-01

Two distinct courses, "Molecular Biology" taught by the Biology Department and "Advanced Biochemistry" taught by the Chemistry Department, complement each other and, when taught in a coordinated and integrated way, can enhance student learning and understanding of complex material. "Molecular Biology" is a comprehensive lecture-based course with a 3-h laboratory once a week, while "Advanced Biochemistry" is a completely laboratory-based course with lecture fully integrated around independent student projects. Both courses emphasize and utilize cutting-edge technology. Teaching across departmental boundaries allows students access to faculty expertise and techniques rarely used at the undergraduate level, namely the tagging of proteins and their use in protein purification. Copyright © 2004 International Union of Biochemistry and Molecular Biology, Inc.

Engineering the ATLAS TAG Browser

NASA Astrophysics Data System (ADS)

Zhang, Qizhi; ATLAS Collaboration

2011-12-01

ELSSI is a web-based event metadata (TAG) browser and event-level selection service for ATLAS. In this paper, we describe some of the challenges encountered in the process of developing ELSSI, and the software engineering strategies adopted to address those challenges. Approaches to management of access to data, browsing, data rendering, query building, query validation, execution, connection management, and communication with auxiliary services are discussed. We also describe strategies for dealing with data that may vary over time, such as run-dependent trigger decision decoding. Along with examples, we illustrate how programming techniques in multiple languages (PHP, JAVASCRIPT, XML, AJAX, and PL/SQL) have been blended to achieve the required results. Finally, we evaluate features of the ELSSI service in terms of functionality, scalability, and performance.

Non-invasive imaging of oxygen extraction fraction in adults with sickle cell anaemia.

PubMed

Jordan, Lori C; Gindville, Melissa C; Scott, Allison O; Juttukonda, Meher R; Strother, Megan K; Kassim, Adetola A; Chen, Sheau-Chiann; Lu, Hanzhang; Pruthi, Sumit; Shyr, Yu; Donahue, Manus J

2016-03-01

Sickle cell anaemia is a monogenetic disorder with a high incidence of stroke. While stroke screening procedures exist for children with sickle cell anaemia, no accepted screening procedures exist for assessing stroke risk in adults. The purpose of this study is to use novel magnetic resonance imaging methods to evaluate physiological relationships between oxygen extraction fraction, cerebral blood flow, and clinical markers of cerebrovascular impairment in adults with sickle cell anaemia. The specific goal is to determine to what extent elevated oxygen extraction fraction may be uniquely present in patients with higher levels of clinical impairment and therefore may represent a candidate biomarker of stroke risk. Neurological evaluation, structural imaging, and the non-invasive T2-relaxation-under-spin-tagging magnetic resonance imaging method were applied in sickle cell anaemia (n = 34) and healthy race-matched control (n = 11) volunteers without sickle cell trait to assess whole-brain oxygen extraction fraction, cerebral blood flow, degree of vasculopathy, severity of anaemia, and presence of prior infarct; findings were interpreted in the context of physiological models. Cerebral blood flow and oxygen extraction fraction were elevated (P < 0.05) in participants with sickle cell anaemia (n = 27) not receiving monthly blood transfusions (interquartile range cerebral blood flow = 46.2-56.8 ml/100 g/min; oxygen extraction fraction = 0.39-0.50) relative to controls (interquartile range cerebral blood flow = 40.8-46.3 ml/100 g/min; oxygen extraction fraction = 0.33-0.38). Oxygen extraction fraction (P < 0.0001) but not cerebral blood flow was increased in participants with higher levels of clinical impairment. These data provide support for T2-relaxation-under-spin-tagging being able to quickly and non-invasively detect elevated oxygen extraction fraction in individuals with sickle cell anaemia with higher levels of clinical impairment. Our results support the premise that magnetic resonance imaging-based assessment of elevated oxygen extraction fraction might be a viable screening tool for evaluating stroke risk in adults with sickle cell anaemia. © The Author (2016). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Non-invasive imaging of oxygen extraction fraction in adults with sickle cell anaemia

PubMed Central

Gindville, Melissa C.; Scott, Allison O.; Juttukonda, Meher R.; Strother, Megan K.; Kassim, Adetola A.; Chen, Sheau-Chiann; Lu, Hanzhang; Pruthi, Sumit; Shyr, Yu; Donahue, Manus J.

2016-01-01

Sickle cell anaemia is a monogenetic disorder with a high incidence of stroke. While stroke screening procedures exist for children with sickle cell anaemia, no accepted screening procedures exist for assessing stroke risk in adults. The purpose of this study is to use novel magnetic resonance imaging methods to evaluate physiological relationships between oxygen extraction fraction, cerebral blood flow, and clinical markers of cerebrovascular impairment in adults with sickle cell anaemia. The specific goal is to determine to what extent elevated oxygen extraction fraction may be uniquely present in patients with higher levels of clinical impairment and therefore may represent a candidate biomarker of stroke risk. Neurological evaluation, structural imaging, and the non-invasive T2-relaxation-under-spin-tagging magnetic resonance imaging method were applied in sickle cell anaemia (n = 34) and healthy race-matched control (n = 11) volunteers without sickle cell trait to assess whole-brain oxygen extraction fraction, cerebral blood flow, degree of vasculopathy, severity of anaemia, and presence of prior infarct; findings were interpreted in the context of physiological models. Cerebral blood flow and oxygen extraction fraction were elevated (P < 0.05) in participants with sickle cell anaemia (n = 27) not receiving monthly blood transfusions (interquartile range cerebral blood flow = 46.2–56.8 ml/100 g/min; oxygen extraction fraction = 0.39–0.50) relative to controls (interquartile range cerebral blood flow = 40.8–46.3 ml/100 g/min; oxygen extraction fraction = 0.33–0.38). Oxygen extraction fraction (P < 0.0001) but not cerebral blood flow was increased in participants with higher levels of clinical impairment. These data provide support for T2-relaxation-under-spin-tagging being able to quickly and non-invasively detect elevated oxygen extraction fraction in individuals with sickle cell anaemia with higher levels of clinical impairment. Our results support the premise that magnetic resonance imaging-based assessment of elevated oxygen extraction fraction might be a viable screening tool for evaluating stroke risk in adults with sickle cell anaemia. PMID:26823369

Laser induced fluorescence spectrum analysis of OH from photo-dissociation of water in gas phase

NASA Astrophysics Data System (ADS)

Li, Guohua; Ye, Jingfeng; Zhang, Zhengrong; Wang, Sheng; Hu, Zhiyun; Zhao, Xinyan

2017-05-01

The OH can be generated from photo-dissociation of water in the gas phase, and the generated OH has served in tagging velocimetry using the time-flight method. The hydroxyl tagging mechanism has the advantages of non-seeding, kindly flow following character, but its application in the reaction region is limited for the fluorescence interference from nascent OH. In this paper, we explored the laser induced fluorescence spectrum of OH both from burning and photodissociation. A photo-dissociation laser induced fluorescence (PD-LIF) system with optical multichannel analysis instrument (OMA) for spectrum analysis was developed. Based on multichannel mechanism, the LIF spectrum of OH from photo-dissociation and burning were acquired simultaneously. The temporal spectrum profiles of dissociation OH both in flame and air were taken by varying the pump-probe delay. The normalized emission spectrum in flame showed a process of rotational relaxation while in air the spectrum was almost not changed. The fluorescence intensity was precisely proportional to the base states population, so we can get certain states that the OH from dissociation was predominant from the fluorescence intensity ratio of OH. This result can be further utilized for hydroxyl tagging velocimetry technology (HTV) which was less affected by burning OH.

Nano-tags for neonates and ocean-mediated swimming behaviours linked to rapid dispersal of hatchling sea turtles

PubMed Central

Scott, Rebecca; Biastoch, Arne; Roder, Christian; Stiebens, Victor A.; Eizaguirre, Christophe

2014-01-01

Dispersal during juvenile life stages drives the life-history evolution and dynamics of many marine vertebrate populations. However, the movements of juvenile organisms, too small to track using conventional satellite telemetry devices, remain enigmatic. For sea turtles, this led to the paradigm of the ‘lost years' since hatchlings disperse widely with ocean currents. Recently, advances in the miniaturization of tracking technology have permitted the application of nano-tags to track cryptic organisms. Here, the novel use of acoustic nano-tags on neonate loggerhead turtle hatchlings enabled us to witness first-hand their dispersal and behaviour during their first day at sea. We tracked hatchlings distances of up to 15 km and documented their rapid transport (up to 60 m min−1) with surface current flows passing their natal areas. Tracking was complemented with laboratory observations to monitor swimming behaviours over longer periods which highlighted (i) a positive correlation between swimming activity levels and body size and (ii) population-specific swimming behaviours (e.g. nocturnal inactivity) suggesting local oceanic conditions drive the evolution of innate swimming behaviours. Knowledge of the swimming behaviours of small organisms is crucial to improve the accuracy of ocean model simulations used to predict the fate of these organisms and determine resultant population-level implications into adulthood. PMID:25339720

Identification of a small molecule yeast TORC1 inhibitor with a multiplex screen based on flow cytometry.

PubMed

Chen, Jun; Young, Susan M; Allen, Chris; Seeber, Andrew; Péli-Gulli, Marie-Pierre; Panchaud, Nicolas; Waller, Anna; Ursu, Oleg; Yao, Tuanli; Golden, Jennifer E; Strouse, J Jacob; Carter, Mark B; Kang, Huining; Bologa, Cristian G; Foutz, Terry D; Edwards, Bruce S; Peterson, Blake R; Aubé, Jeffrey; Werner-Washburne, Margaret; Loewith, Robbie J; De Virgilio, Claudio; Sklar, Larry A

2012-04-20

TOR (target of rapamycin) is a serine/threonine kinase, evolutionarily conserved from yeast to human, which functions as a fundamental controller of cell growth. The moderate clinical benefit of rapamycin in mTOR-based therapy of many cancers favors the development of new TOR inhibitors. Here we report a high-throughput flow cytometry multiplexed screen using five GFP-tagged yeast clones that represent the readouts of four branches of the TORC1 signaling pathway in budding yeast. Each GFP-tagged clone was differentially color-coded, and the GFP signal of each clone was measured simultaneously by flow cytometry, which allows rapid prioritization of compounds that likely act through direct modulation of TORC1 or proximal signaling components. A total of 255 compounds were confirmed in dose-response analysis to alter GFP expression in one or more clones. To validate the concept of the high-throughput screen, we have characterized CID 3528206, a small molecule most likely to act on TORC1 as it alters GFP expression in all five GFP clones in a manner analogous to that of rapamycin. We have shown that CID 3528206 inhibited yeast cell growth and that CID 3528206 inhibited TORC1 activity both in vitro and in vivo with EC(50)'s of 150 nM and 3.9 μM, respectively. The results of microarray analysis and yeast GFP collection screen further support the notion that CID 3528206 and rapamycin modulate similar cellular pathways. Together, these results indicate that the HTS has identified a potentially useful small molecule for further development of TOR inhibitors.

Influence of Yo-Yo IR2 Scores on Internal and External Workloads and Fatigue Responses of Tag Football Players during Tournament Competition

PubMed Central

2015-01-01

The purpose of this study was to: a) identify changes in jump height and perceived well-being as indirect markers of fatigue, b) determine the internal and external workloads performed by players, and c) examine the influence of Yo-Yo IR2 on changes in jump height, perceived well-being and internal and external workloads during a tag football tournament. Microtechnology devices combined with heart rate (HR) chest straps provided external and internal measures of match work-rate and workload for twelve male tag football players during the 2014 Australian National Championships. Jump height and perceived well-being were assessed prior to and during the tournament as indirect measures of fatigue. Changes in work-rate, workload and fatigue measures between high- and low-fitness groups were examined based on players’ Yo-Yo IR2 score using a median split technique. The low- and high-fitness groups reported similar mean HR, PlayerloadTM/min, and distance/min for matches, however the low-fitness group reported higher perceived match-intensities (ES = 0.90–1.35) for several matches. Further, the high-fitness group reported higher measures of tournament workload, including distance (ES = 0.71), PlayerloadTM (ES = 0.85) and Edwards’ training impulse (TRIMP) (ES = 1.23) than the low-fitness group. High- and low-fitness groups both showed large decreases (ES = 1.46–1.49) in perceived well-being during the tournament, although jump height did not decrease below pre-tournament values. Increased Yo-Yo IR2 appears to offer a protective effect against player fatigue despite increased workloads during a tag football tournament. It is vital that training programs adequately prepare tag football players for tournament competition to maximise performance and minimise player fatigue. PMID:26465599

One-step methodology for the direct covalent capture of GPCRs from complex matrices onto solid surfaces based on the bioorthogonal reaction between haloalkane dehalogenase and chloroalkanes† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c7sc03887a

PubMed Central

Zeng, Kaizhu; Li, Qian; Wang, Jing; Yin, Guowei; Zhang, Yajun; Xiao, Chaoni; Fan, Taiping; Zheng, Xiaohui

2017-01-01

Protein immobilization techniques play an important role in the development of assays for disease diagnosis and drug discovery. However, many of these approaches are not applicable to transmembrane proteins. G protein-coupled receptors (GPCRs) are the largest protein superfamily encoded by the human genome and are targeted by a quarter of all prescription drugs. GPCRs are highly dynamic and sensitive to changes in the ambient environment, and current immobilization methodologies are not suitable for GPCRs. We used haloalkane dehalogenase (Halo) as an immobilization tag fused to the β2-adrenoceptor (β2-AR), angiotensin II type 1 (AT1) and angiotensin II type 2 (AT2) receptors. The engineered Halo-tag covalently binds to a specific substrate chloroalkane through Asp 106 in the catalytic pocket. The Halo-tagged GPCRs were expressed in Escherichia coli at a suitable yield. Accordingly, we loaded cell lysate containing Halo-tagged GPCRs onto a macroporous silica gel coated with chloroalkane. Morphological characterization indicated a homogeneous monolayer of immobilized Halo-tagged GPCRs on the silica gel surface. The immobilized receptors proved to be surrounded by specific bound phospholipids including PG C18:1/C18:1. We observed a radio-ligand binding ability and ligand-induced conformational changes in the immobilized GPCRs, suggesting the preservation of bioactivity. This method is a one-step approach for the specific immobilization of GPCRs from cell lysates and validates that immobilized receptors retain canonical ligand binding capacity. Our immobilization strategy circumvents labor-intensive purification procedures and minimizes loss of activity. The immobilized receptors can be applied to high-throughput drug and interaction partner screening for GPCRs. PMID:29629116

Generation and analysis of a barcode-tagged insertion mutant library in the fission yeast Schizosaccharomyces pombe.

PubMed

Chen, Bo-Ruei; Hale, Devin C; Ciolek, Peter J; Runge, Kurt W

2012-05-03

Barcodes are unique DNA sequence tags that can be used to specifically label individual mutants. The barcode-tagged open reading frame (ORF) haploid deletion mutant collections in the budding yeast Saccharomyces cerevisiae and the fission yeast Schizosaccharomyces pombe allow for high-throughput mutant phenotyping because the relative growth of mutants in a population can be determined by monitoring the proportions of their associated barcodes. While these mutant collections have greatly facilitated genome-wide studies, mutations in essential genes are not present, and the roles of these genes are not as easily studied. To further support genome-scale research in S. pombe, we generated a barcode-tagged fission yeast insertion mutant library that has the potential of generating viable mutations in both essential and non-essential genes and can be easily analyzed using standard molecular biological techniques. An insertion vector containing a selectable ura4+ marker and a random barcode was used to generate a collection of 10,000 fission yeast insertion mutants stored individually in 384-well plates and as six pools of mixed mutants. Individual barcodes are flanked by Sfi I recognition sites and can be oligomerized in a unique orientation to facilitate barcode sequencing. Independent genetic screens on a subset of mutants suggest that this library contains a diverse collection of single insertion mutations. We present several approaches to determine insertion sites. This collection of S. pombe barcode-tagged insertion mutants is well-suited for genome-wide studies. Because insertion mutations may eliminate, reduce or alter the function of essential and non-essential genes, this library will contain strains with a wide range of phenotypes that can be assayed by their associated barcodes. The design of the barcodes in this library allows for barcode sequencing using next generation or standard benchtop cloning approaches.

Free-living bacterial communities associated with tubeworm (Ridgeia piscesae) aggregations in contrasting diffuse flow hydrothermal vent habitats at the Main Endeavour Field, Juan de Fuca Ridge

PubMed Central

Forget, Nathalie L; Kim Juniper, S

2013-01-01

We systematically studied free-living bacterial diversity within aggregations of the vestimentiferan tubeworm Ridgeia piscesae sampled from two contrasting flow regimes (High Flow and Low Flow) in the Endeavour Hydrothermal Vents Marine Protected Area (MPA) on the Juan de Fuca Ridge (Northeast Pacific). Eight samples of particulate detritus were recovered from paired tubeworm grabs from four vent sites. Most sequences (454 tag and Sanger methods) were affiliated to the Epsilonproteobacteria, and the sulfur-oxidizing genus Sulfurovum was dominant in all samples. Gammaproteobacteria were also detected, mainly in Low Flow sequence libraries, and were affiliated with known methanotrophs and decomposers. The cooccurrence of sulfur reducers from the Deltaproteobacteria and the Epsilonproteobacteria suggests internal sulfur cycling within these habitats. Other phyla detected included Bacteroidetes, Actinobacteria, Chloroflexi, Firmicutes, Planctomycetes, Verrucomicrobia, and Deinococcus–Thermus. Statistically significant relationships between sequence library composition and habitat type suggest a predictable pattern for High Flow and Low Flow environments. Most sequences significantly more represented in High Flow libraries were related to sulfur and hydrogen oxidizers, while mainly heterotrophic groups were more represented in Low Flow libraries. Differences in temperature, available energy for metabolism, and stability between High Flow and Low Flow habitats potentially explain their distinct bacterial communities. PMID:23401293

Development of Optimized Combustors and Thermoelectric Generators for Palm Power Generation

DTIC Science & Technology

2004-10-26

manufacturing techniques and microfabrication, on the chemical kinetics of JP-8 surrogates and on the development of advanced laser diagnostics for JP-8...takes the shape of a cone from the tip of which a thin liquid thread emerges, in the so-called cone-jet mode [1]. This microjet breaks into a stream of...combustion systems. 2. The development of a diagnostic technique based on two-color laser induced fluorescence from fluorescence tags added to the fuel

Top quark studies at hadron colliders

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

Sinervo, P.K.

1997-01-01

The techniques used to study top quarks at hadron colliders are presented. The analyses that discovered the top quark are described, with emphasis on the techniques used to tag b quark jets in candidate events. The most recent measurements of top quark properties by the CDF and DO Collaborations are reviewed, including the top quark cross section, mass, branching fractions, and production properties. Future top quark studies at hadron colliders are discussed, and predictions for event yields and uncertainties in the measurements of top quark properties are presented.

Comparison of three nonlinear models to describe long-term tag shedding by lake trout

USGS Publications Warehouse

Fabrizio, Mary C.; Swanson, Bruce L.; Schram, Stephen T.; Hoff, Michael H.

1996-01-01

We estimated long-term tag-shedding rates for lake trout Salvelinus namaycush using two existing models and a model we developed to account for the observed permanence of some tags. Because tag design changed over the course of the study, we examined tag-shedding rates for three types of numbered anchor tags (Floy tags FD-67, FD-67C, and FD-68BC) and an unprinted anchor tag (FD-67F). Lake trout from the Gull Island Shoal region, Lake Superior, were double-tagged, and subsequent recaptures were monitored in annual surveys conducted from 1974 to 1992. We modeled tag-shedding rates, using time at liberty and probabilities of tag shedding estimated from fish released in 1974 and 1978–1983 and later recaptured. Long-term shedding of numbered anchor tags in lake trout was best described by a nonlinear model with two parameters: an instantaneous tag-shedding rate and a constant representing the proportion of tags that were never shed. Although our estimates of annual shedding rates varied with tag type (0.300 for FD-67, 0.441 for FD-67C, and 0.656 for FD-68BC), differences were not significant. About 36% of tags remained permanently affixed to the fish. Of the numbered tags that were shed (about 64%), two mechanisms contributed to tag loss: disintegration and dislodgment. Tags from about 11% of recaptured fish had disintegrated, but most tags were dislodged. Unprinted tags were shed at a significant but low rate immediately after release, but the long-term, annual shedding rate of these tags was only 0.013. Compared with unprinted tags, numbered tags dislodged at higher annual rates; we hypothesized that this was due to the greater frictional drag associated with the larger cross-sectional area of numbered tags.

Smolt Monitoring at the Head of Lower Granite Reservoir and Lower Granite Dam, 2002 Annual Report.

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

Buettner, Edwin W.; Putnam, Scott A.

This project monitored the daily passage of Chinook salmon Oncorhynchus tshawytscha, steelhead trout O. mykiss, and sockeye salmon smolts O. nerka during the 2002 spring out-migration at migrant traps on the Snake River and Salmon River. In 2002 fish management agencies released significant numbers of hatchery Chinook salmon and steelhead trout above Lower Granite Dam that were not marked with a fin clip or coded-wire tag. Generally, these fish were distinguishable from wild fish by the occurrence of fin erosion. Total annual hatchery Chinook salmon catch at the Snake River trap was 11.4 times greater in 2002 than in 2001.more » The wild Chinook catch was 15.5 times greater than the previous year. Hatchery steelhead trout catch was 2.9 times greater than in 2001. Wild steelhead trout catch was 2.8 times greater than the previous year. The Snake River trap collected 3,996 age-0 Chinook salmon of unknown rearing. During 2002, the Snake River trap captured 69 hatchery and 235 wild/natural sockeye salmon and 114 hatchery coho salmon O. kisutch. Differences in trap catch between years are due to fluctuations not only in smolt production, but also differences in trap efficiency and duration of trap operation associated with flow. The significant increase in catch in 2002 was due to a 3.1 fold increase in hatchery Chinook production and a more normal spring runoff. Trap operations began on March 10 and were terminated on June 7. The trap was out of operation for a total of four days due to mechanical failure or debris. Hatchery Chinook salmon catch at the Salmon River trap was 4.2 times greater and wild Chinook salmon catch was 2.4 times greater than in 2001. The hatchery steelhead trout collection in 2002 was 81% of the 2001 numbers. Wild steelhead trout collection in 2002 was 81% of the previous year's catch. Trap operations began on March 10 and were terminated on May 29 due to high flows. The trap was out of operation for four days due to high flow or debris. The increase in hatchery Chinook catch in 2002 was due to a 3.1 fold increase in hatchery production and differences in flow between years. Changes in hatchery and wild steelhead catch are probably due to differences in flow between years. Travel time (d) and migration rate (km/d) through Lower Granite Reservoir for PIT-tagged Chinook salmon and steelhead trout marked at the Snake River trap were affected by discharge. Statistical analysis of 2002 data detected a relation between migration rate and discharge for hatchery and wild Chinook salmon. For hatchery and wild Chinook salmon there was a 4.7-fold and a 3.7-fold increase in migration rate, respectively, between 50 and 100 kcfs. For steelhead trout tagged at the Snake River trap, statistical analysis detected a significant relation between migration rate and Lower Granite Reservoir inflow discharge. For hatchery and wild steelhead trout, there was a 1.8-fold and a 1.7-fold increase in migration rate, respectively, between 50 and 100 kcfs. Travel time and migration rate to Lower Granite Dam for fish marked at the Salmon River trap were calculated. Statistical analysis of the 2002 data detected a significant relation between migration rate and Lower Granite Reservoir inflow discharge for wild Chinook salmon and hatchery steelhead trout. The analysis was unable to detect a relation between migration rate and discharge for hatchery Chinook salmon. The lack of a detectable relation was probably a result of the migration rate data being spread over a very narrow range of discharge. Not enough data were available to perform the analysis for wild steelhead trout. Migration rate increased 4.3-fold for wild Chinook salmon and 2.2-fold for hatchery steelhead between 50 kcfs and 100 kcfs. Fish tagged with passive integrated transponder (PIT) tags at the Snake River trap were interrogated at four dams with PIT tag detection systems (Lower Granite, Little Goose, Lower Monumental, and McNary dams). Because of the addition of the fourth interrogation site (Lower Monumental) in 1993 and the installation of the Removable Spillway Weir at Lower Granite Dam in 2000, caution must be used in comparing cumulative interrogation data. Cumulative interrogations at the four dams for fish marked at the Snake River trap were 61% for hatchery Chinook, 68% for wild Chinook, 58% for hatchery steelhead, and 62% for wild steelhead. Cumulative interrogations at the four dams for fish marked at the Salmon River trap were 51% for hatchery Chinook, 59% for wild Chinook salmon, 45% for hatchery steelhead trout, and 54% for wild steelhead trout. Cumulative interrogations were significantly lower in 2002 than in previous years with similar flow.« less

Emigration of Natural and Hatchery Chinook Salmon and Steelhead Smolts from the Imnaha River, Oregon, 1998-1999 Annual Report.

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

Cleary, Peter J.; Blenden, Michael L.; Kucera, Paul A.

2002-08-01

This report summarizes the results of the Lower Snake River Compensation Plan Hatchery Evaluation Studies (LSRCP) and the Imnaha Smolt Monitoring Program (SMP) for the 1999 smolt migration from the Imnaha River, Oregon. These studies were designed and closely coordinated to provide information about juvenile natural and hatchery chinook salmon (Oncorhynchus tshawytscha) and steelhead (Oncorhynchus mykiss) biological characteristics, behavior and emigrant timing, survival, arrival timing and travel time to the Snake River dams and McNary Dam on the Columbia River. Data collected from these studies are shared with the Fish Passage Center (FPC). These data are essential to quantify smoltmore » survival rates under the current passage conditions and to evaluate the future recovery strategies that seek to optimize smolt survival through the hydroelectric system. Information shared with the FPC assists with in-season shaping of flow and spill management requests in the Snake River reservoirs. The Bonneville Power Administration and the United States Fish and Wildlife Service contracted the Nez Perce Tribe (NPT) to monitor emigration timing and tag 21,200 emigrating natural and hatchery chinook salmon and steelhead smolts from the Imnaha River during the spring emigration period (March 1-June 15) with passive integrated transponder (PIT) tags. The completion of trapping in the spring of 1999 marked the eighth year of emigration studies on the Imnaha River and the sixth year of participating in the FPC smolt monitoring program. Monitoring and evaluation objectives were to: (1) Determine spring emigration timing of chinook salmon and steelhead smolts collected at the Imnaha River trap. (2) Evaluate effects of flow, temperature and other environmental factors on emigration timing. (3) Monitor the daily catch and biological characteristics of juvenile chinook salmon and steelhead smolts collected at the Imnaha River screw trap. (4) Determine emigration timing, travel time, and in-river survival of PIT tagged hatchery chinook salmon smolts released at the Imnaha River acclimation facility to the Imnaha River Trap. (5) Determine arrival timing, travel time and estimated survival of PIT tagged hatchery and natural chinook salmon and natural and hatchery steelhead smolts from the Imnaha River to Snake and Columbia river dams.« less

Visualizing Time-Varying Phenomena In Numerical Simulations Of Unsteady Flows

NASA Technical Reports Server (NTRS)

Lane, David A.

1996-01-01

Streamlines, contour lines, vector plots, and volume slices (cutting planes) are commonly used for flow visualization. These techniques are sometimes referred to as instantaneous flow visualization techniques because calculations are based on an instant of the flowfield in time. Although instantaneous flow visualization techniques are effective for depicting phenomena in steady flows,they sometimes do not adequately depict time-varying phenomena in unsteady flows. Streaklines and timelines are effective visualization techniques for depicting vortex shedding, vortex breakdown, and shock waves in unsteady flows. These techniques are examples of time-dependent flow visualization techniques, which are based on many instants of the flowfields in time. This paper describes the algorithms for computing streaklines and timelines. Using numerically simulated unsteady flows, streaklines and timelines are compared with streamlines, contour lines, and vector plots. It is shown that streaklines and timelines reveal vortex shedding and vortex breakdown more clearly than instantaneous flow visualization techniques.

Inductively Coupled Plasma Mass Spectrometry (ICP-MS) Applications in Quantitative Proteomics.

PubMed

Chahrour, Osama; Malone, John

2017-01-01

Recent advances in inductively coupled plasma mass spectrometry (ICP-MS) hyphenated to different separation techniques have promoted it as a valuable tool in protein/peptide quantification. These emerging ICP-MS applications allow absolute quantification by measuring specific elemental responses. One approach quantifies elements already present in the structure of the target peptide (e.g. phosphorus and sulphur) as natural tags. Quantification of these natural tags allows the elucidation of the degree of protein phosphorylation in addition to absolute protein quantification. A separate approach is based on utilising bi-functional labelling substances (those containing ICP-MS detectable elements), that form a covalent chemical bond with the protein thus creating analogs which are detectable by ICP-MS. Based on the previously established stoichiometries of the labelling reagents, quantification can be achieved. This technique is very useful for the design of precise multiplexed quantitation schemes to address the challenges of biomarker screening and discovery. This review discusses the capabilities and different strategies to implement ICP-MS in the field of quantitative proteomics. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Subdiffractional tracking of internalized molecules reveals heterogeneous motion states of synaptic vesicles

PubMed Central

Morrow, Isabel C.; Harper, Callista B.

2016-01-01

Our understanding of endocytic pathway dynamics is severely restricted by the diffraction limit of light microscopy. To address this, we implemented a novel technique based on the subdiffractional tracking of internalized molecules (sdTIM). This allowed us to image anti–green fluorescent protein Atto647N-tagged nanobodies trapped in synaptic vesicles (SVs) from live hippocampal nerve terminals expressing vesicle-associated membrane protein 2 (VAMP2)–pHluorin with 36-nm localization precision. Our results showed that, once internalized, VAMP2–pHluorin/Atto647N–tagged nanobodies exhibited a markedly lower mobility than on the plasma membrane, an effect that was reversed upon restimulation in presynapses but not in neighboring axons. Using Bayesian model selection applied to hidden Markov modeling, we found that SVs oscillated between diffusive states or a combination of diffusive and transport states with opposite directionality. Importantly, SVs exhibiting diffusive motion were relatively less likely to switch to the transport motion. These results highlight the potential of the sdTIM technique to provide new insights into the dynamics of endocytic pathways in a wide variety of cellular settings. PMID:27810917

Imaging intracellular protein dynamics by spinning disk confocal microscopy

PubMed Central

Stehbens, Samantha; Pemble, Hayley; Murrow, Lindsay; Wittmann, Torsten

2012-01-01

The palette of fluorescent proteins has grown exponentially over the last decade, and as a result live imaging of cells expressing fluorescently tagged proteins is becoming more and more main stream. Spinning disk confocal microscopy (SDC) is a high speed optical sectioning technique, and a method of choice to observe and analyze intracellular fluorescent protein dynamics at high spatial and temporal resolution. In an SDC system, a rapidly rotating pinhole disk generates thousands of points of light that scan the specimen simultaneously, which allows direct capture of the confocal image with low noise scientific grade cooled charged-coupled device (CCD) cameras, and can achieve frame rates of up 1000 frames per second. In this chapter we describe important components of a state-of-the-art spinning disk system optimized for live cell microscopy, and provide a rationale for specific design choices. We also give guidelines how other imaging techniques such as total internal reflection (TIRF) microscopy or spatially controlled photoactivation can be coupled with SDC imaging, and provide a short protocol on how to generate cell lines stably expressing fluorescently tagged proteins by lentivirus-mediated transduction. PMID:22264541

Use of LysoTracker dyes: a flow cytometric study of autophagy.

PubMed

Chikte, Shaheen; Panchal, Neelam; Warnes, Gary

2014-02-01

The flow cytometric use of LysoTracker dyes was employed to investigate the autophagic process and to compare this with the upregulation of autophagy marker, the microtubule-associated protein LC3B. Although the mechanism of action of LysoTracker dyes is not fully understood, they have been used in microscopy to image acidic spherical organelles, and their use in flow cytometry has not been thoroughly investigated in the study of autophagy. This investigation uses numerous autophagy-inducing agents including chloroquine (CQ), rapamycin, low serum (<1%) RPMI, and nutrient starvation to induce autophagy in Jurkat T-cell leukemia and K562 erythromyeloid cell lines. LC3B showed an increase with CQ treatment although this was different to LysoTracker signals in terms of dose and time. Rapamycin, low serum (<1%) RPMI, and nutrient starvation induction of autophagy also induced an increase in LysoTracker and LC3B signals. CQ also induced apoptosis in cell lines, which was blocked by pan-caspase inhibitor z-VAD resulting in a reduction in cells undergoing apoptosis and a subsequent upregulation of autophagic markers LC3B and lysosomal dye signals. Given that LC3B and LysoTracker are measuring different biological events in the autophagic process, they surprisingly both upregulated during autophagic process. This study, however, shows that although LysoTracker dyes do not specifically label lysosomes or autophagosomes within the cell, they allow the simultaneous measurement of an autophagy-related process and other live-cell functions, which are not possible with the standard LC3B antibody-labeling technique. This method has the advantage of other live-cell LCB-GFP-tagged experiments in that be used to analyze patient cells as well as easier to use and significantly less costly. Copyright © 2013 International Society for Advancement of Cytometry.

Cellular automaton formulation of passive scalar dynamics

NASA Technical Reports Server (NTRS)

Chen, Hudong; Matthaeus, William H.

1987-01-01

Cellular automata modeling of the advection of a passive scalar in a two-dimensional flow is examined in the context of discrete lattice kinetic theory. It is shown that if the passive scalar is represented by tagging or 'coloring' automation particles a passive advection-diffusion equation emerges without use of perturbation expansions. For the specific case of the hydrodynamic lattice gas model of Frisch et al. (1986), the diffusion coefficient is calculated by perturbation.

Method for rapid base sequencing in DNA and RNA

DOEpatents

Jett, J.H.; Keller, R.A.; Martin, J.C.; Moyzis, R.K.; Ratliff, R.L.; Shera, E.B.; Stewart, C.C.

1987-10-07

A method is provided for the rapid base sequencing of DNA or RNA fragments wherein a single fragment of DNA or RNA is provided with identifiable bases and suspended in a moving flow stream. An exonuclease sequentially cleaves individual bases from the end of the suspended fragment. The moving flow stream maintains the cleaved bases in an orderly train for subsequent detection and identification. In a particular embodiment, individual bases forming the DNA or RNA fragments are individually tagged with a characteristic fluorescent dye. The train of bases is then excited to fluorescence with an output spectrum characteristic of the individual bases. Accordingly, the base sequence of the original DNA or RNA fragment can be reconstructed. 2 figs.

Method for rapid base sequencing in DNA and RNA

DOEpatents

Jett, J.H.; Keller, R.A.; Martin, J.C.; Moyzis, R.K.; Ratliff, R.L.; Shera, E.B.; Stewart, C.C.

1990-10-09

A method is provided for the rapid base sequencing of DNA or RNA fragments wherein a single fragment of DNA or RNA is provided with identifiable bases and suspended in a moving flow stream. An exonuclease sequentially cleaves individual bases from the end of the suspended fragment. The moving flow stream maintains the cleaved bases in an orderly train for subsequent detection and identification. In a particular embodiment, individual bases forming the DNA or RNA fragments are individually tagged with a characteristic fluorescent dye. The train of bases is then excited to fluorescence with an output spectrum characteristic of the individual bases. Accordingly, the base sequence of the original DNA or RNA fragment can be reconstructed. 2 figs.

Method for rapid base sequencing in DNA and RNA

DOEpatents

Jett, James H.; Keller, Richard A.; Martin, John C.; Moyzis, Robert K.; Ratliff, Robert L.; Shera, E. Brooks; Stewart, Carleton C.

1990-01-01

A method is provided for the rapid base sequencing of DNA or RNA fragments wherein a single fragment of DNA or RNA is provided with identifiable bases and suspended in a moving flow stream. An exonuclease sequentially cleaves individual bases from the end of the suspended fragment. The moving flow stream maintains the cleaved bases in an orderly train for subsequent detection and identification. In a particular embodiment, individual bases forming the DNA or RNA fragments are individually tagged with a characteristic fluorescent dye. The train of bases is then excited to fluorescence with an output spectrum characteristic of the individual bases. Accordingly, the base sequence of the original DNA or RNA fragment can be reconstructed.

High affinity receptor labeling based on basic leucine zipper domain peptides conjugated with pH-sensitive fluorescent dye: Visualization of AMPA-type glutamate receptor endocytosis in living neurons.

PubMed

Hayashi, Ayako; Asanuma, Daisuke; Kamiya, Mako; Urano, Yasuteru; Okabe, Shigeo

2016-01-01

Techniques to visualize receptor trafficking in living neurons are important, but currently available methods are limited in their labeling efficiency, specificity and reliability. Here we report a method for receptor labeling with a basic leucine zipper domain peptide (ZIP) and a binding cassette specific to ZIP. Receptors are tagged with a ZIP-binding cassette at their extracellular domain. Tagged receptors expressed in cultured cells were labeled with exogenously applied fluorescently labeled ZIP with low background and high affinity. To test if ZIP labeling is useful in monitoring endocytosis and intracellular trafficking, we next conjugated ZIP with a pH-sensitive dye RhP-M (ZIP-RhP-M). ZIP binding to its binding cassette was pH-resistant and RhP-M fluorescence dramatically increased in acidic environment. Thus AMPA-type glutamate receptors (AMPARs) labeled by ZIP-RhP-M can report receptor endocytosis and subsequent intracellular trafficking. Application of ZIP-RhP-M to cultured hippocampal neurons expressing AMPARs tagged with a ZIP-binding cassette resulted in appearance of fluorescent puncta in PSD-95-positive large spines, suggesting local endocytosis and acidification of AMPARs in individual mature spines. This spine pool of AMPARs in acidic environment was distinct from the early endosomes labeled by transferrin uptake. These results suggest that receptor labeling by ZIP-RhP-M is a useful technique for monitoring endocytosis and intracellular trafficking. This article is part of the Special Issue entitled 'Synaptopathy--from Biology to Therapy'. Copyright © 2015 Elsevier Ltd. All rights reserved.

Two-Way Communication Using RFID Equipment and Techniques

NASA Technical Reports Server (NTRS)

Jedry, Thomas; Archer, Eric

2007-01-01

Equipment and techniques used in radio-frequency identification (RFID) would be extended, according to a proposal, to enable short-range, two-way communication between electronic products and host computers. In one example of a typical contemplated application, the purpose of the short-range radio communication would be to transfer image data from a user s digital still or video camera to the user s computer for recording and/or processing. The concept is also applicable to consumer electronic products other than digital cameras (for example, cellular telephones, portable computers, or motion sensors in alarm systems), and to a variety of industrial and scientific sensors and other devices that generate data. Until now, RFID has been used to exchange small amounts of mostly static information for identifying and tracking assets. Information pertaining to an asset (typically, an object in inventory to be tracked) is contained in miniature electronic circuitry in an RFID tag attached to the object. Conventional RFID equipment and techniques enable a host computer to read data from and, in some cases, to write data to, RFID tags, but they do not enable such additional functions as sending commands to, or retrieving possibly large quantities of dynamic data from, RFID-tagged devices. The proposal would enable such additional functions. The figure schematically depicts an implementation of the proposal for a sensory device (e.g., a digital camera) that includes circuitry that converts sensory information to digital data. In addition to the basic sensory device, there would be a controller and a memory that would store the sensor data and/or data from the controller. The device would also be equipped with a conventional RFID chipset and antenna, which would communicate with a host computer via an RFID reader. The controller would function partly as a communication interface, implementing two-way communication protocols at all levels (including RFID if needed) between the sensory device and the memory and between the host computer and the memory. The controller would perform power V

Acousto-optical imaging using a powerful long pulse laser

NASA Astrophysics Data System (ADS)

Rousseau, Guy; Blouin, Alain; Monchalin, Jean-Pierre

2008-06-01

Acousto-optical imaging is an emerging biodiagnostic technique which provides an optical spectroscopic signature and a spatial localization of an optically absorbing target embedded in a strongly scattering medium. The transverse resolution of the technique is determined by the lateral extent of ultrasound beam focal zone while the axial resolution is obtained by using short ultrasound pulses. Although very promising for medical diagnostic, the practical application of this technique is presently limited by its poor sensitivity. Moreover, any method to enhance the signal-to-noise ratio must obviously satisfy the in vivo safety limits regarding the acceptable power level of both the ultrasonic pressure wave and the laser beam. In this paper, we propose to improve the sensitivity by using a pulsed single-frequency laser source to raise the optical peak power applied to the scattering medium and to collect more ultrasonically tagged photons. Such a laser source also allows illuminating the tissues mainly during the transit time of the ultrasonic wave to maintain the average optical power below the maximum permissible exposure. In our experiment, a single-frequency Nd:YAG laser emitting 500-μs pulses with a peak power superior to 100 W was used. Photons were tagged in few-cm thick optical phantoms with tone bursts generated by an ultrasonic transducer. Tagged photons were detected with a GaAs photorefractive interferometer characterized by a large optical etendue to process simultaneously a large number of speckle grains. When pumped by high intensity laser pulses, such an interferometer also provides the fast response time essential to obtain an apparatus insensitive to the speckle decorrelation due to mechanical vibrations or tissues movements. The use of a powerful long pulse laser appears promising to enhance the signal level in ultrasound modulated optical imaging. When combined with a photorefractive interferometer of large optical etendue, such a source could allow obtaining both the sensitivity and the fast response time necessary for biodiagnostic applications.

System and method for acquisition management of subject position information

DOEpatents

Carrender, Curt

2005-12-13

A system and method for acquisition management of subject position information that utilizes radio frequency identification (RF ID) to store position information in position tags. Tag programmers receive position information from external positioning systems, such as the Global Positioning System (GPS), from manual inputs, such as keypads, or other tag programmers. The tag programmers program each position tag with the received position information. Both the tag programmers and the position tags can be portable or fixed. Implementations include portable tag programmers and fixed position tags for subject position guidance, and portable tag programmers for collection sample labeling. Other implementations include fixed tag programmers and portable position tags for subject route recordation. Position tags can contain other associated information such as destination address of an affixed subject for subject routing.

System and method for acquisition management of subject position information

DOEpatents

Carrender, Curt [Morgan Hill, CA

2007-01-23

A system and method for acquisition management of subject position information that utilizes radio frequency identification (RF ID) to store position information in position tags. Tag programmers receive position information from external positioning systems, such as the Global Positioning System (GPS), from manual inputs, such as keypads, or other tag programmers. The tag programmers program each position tag with the received position information. Both the tag programmers and the position tags can be portable or fixed. Implementations include portable tag programmers and fixed position tags for subject position guidance, and portable tag programmers for collection sample labeling. Other implementations include fixed tag programmers and portable position tags for subject route recordation. Position tags can contain other associated information such as destination address of an affixed subject for subject routing.

A laboratory evaluation of tagging-related mortality and tag loss in juvenile humpback chub

USGS Publications Warehouse

Ward, David L.; Persons, William R.; Young, Kirk; Stone, Dennis M.; Van Haverbeke, Randy; Knight, William R.

2015-01-01

We quantified tag retention, survival, and growth in juvenile, captive-reared Humpback Chub Gila cypha marked with three different tag types: (1) Biomark 12.5-mm, 134.2-kHz, full duplex PIT tags injected into the body cavity with a 12-gauge needle; (2) Biomark 8.4-mm, 134.2-kHz, full duplex PIT tags injected with a 16-gauge needle; and (3) Northwest Marine Technology visible implant elastomer (VIE) tags injected under the skin with a 29-gauge needle. Estimates of tag loss, tagging-induced mortality, and growth were evaluated for 60 d with each tag type for four different size-groups of fish: 40–49 mm, 50–59 mm, 60–69 mm, and 70–79 mm TL. Total length was a significant predictor of the probability of PIT tag retention and mortality for both 8-mm and 12-mm PIT tags, and the smallest fish had the highest rates of tag loss (12.5–30.0%) and mortality (7.5–20.0%). Humpback Chub of sizes 40–49 mm TL and tagged with VIE tags had no mortality but did have a 17.5% tag loss. Growth rates of all tagged fish were similar to controls. Our data indicate Humpback Chub can be effectively tagged using either 8-mm or 12-mm PIT tags with little tag loss or mortality at sizes as low as 65 mm TL.

Regional cardiac function analysis from tagged MRI images. Comparison of techniques: Harmonic-Phase (HARP) versus Sinusoidal-Modeling (SinMod) analysis.

PubMed

Ibrahim, El-Sayed H; Stojanovska, Jadranka; Hassanein, Azza; Duvernoy, Claire; Croisille, Pierre; Pop-Busui, Rodica; Swanson, Scott D

2018-05-16

Cardiac MRI tagging is a valuable technique for evaluating regional heart function. Currently, there are a number of different techniques for analyzing the tagged images. Specifically, k-space-based analysis techniques showed to be much faster than image-based techniques, where harmonic-phase (HARP) and sine-wave modeling (SinMod) stand as two famous techniques of the former group, which are frequently used in clinical studies. In this study, we compared HARP and SinMod and studied inter-observer variability between the two techniques for evaluating myocardial strain and apical-to-base torsion in numerical phantom, nine healthy controls, and thirty diabetic patients. Based on the ground-truth numerical phantom measurements (strain = -20% and rotation angle = -4.4°), HARP and SinMod resulted in overestimation (in absolute value terms) of strain by 1% and 5% (strain values), and of rotation angle by 0.4° and 2.0°, respectively. For the in-vivo results, global strain and torsion ranges were -10.6 to -35.3% and 1.8-12.7°/cm in patients, and -17.8 to -32.7% and 1.8-12.3°/cm in volunteers. On average, SinMod overestimated strain measurements by 5.7% and 5.9% (strain values) in the patients and volunteers, respectively, compared to HARP, and overestimated torsion measurements by 2.9°/cm and 2.5°/cm in the patients and volunteers, respectively, compared to HARP. Location-wise, the ranges for basal, mid-ventricular, and apical strain in patients (volunteers) were -8.4 to -31.5% (-11.6 to -33.3%), -6.3 to -37.2% (-17.8 to -33.3%), and -5.2 to -38.4% (-20.0 to -33.2%), respectively. SinMod overestimated strain in the basal, mid-ventricular, and apical slices by 4.7% (5.7%), 5.9% (5.5%), and 8.9% (6.8%), respectively, compared to HARP in the patients (volunteers). Nevertheless, there existed good correlation between the HARP and SinMod measurements. Finally, there were no significant strain or torsion measurement differences between patients and volunteers. There existed good inter-observer agreement, as all measurement differences lied within the Bland-Altman ± 2 standard-deviation (SD) difference limits. In conclusion, despite the consistency of the results by either HARP or SinMod and acceptable agreement of the generated strain and torsion patterns by both techniques, SinMod systematically overestimated the measurements compared to HARP. Under current operating conditions, the measurements from HARP and SinMod cannot be used interchangeably. Copyright © 2017. Published by Elsevier Inc.

The Means: Cytometry and Mass Spectrometry Converge in a Single Cell Deep Profiling Platform

PubMed Central

Weis-Garcia, Frances; Bandura, Dmitry; Baranov, Vladimir; Ornatsky, Olga; Tanner, Scott

2013-01-01

Inductively Coupled Plasma Mass Spectrometry (ICP-MS) is a distinct flavor of mass spectrometry that has had little association with cell biology: it remains the state of the art for the determination of the atomic composition of materials. Unrelatedly, flow cytometry is the superior method for distinguishing the heterogeneity of cells through the determination of antigen signatures using tagged antibodies. Simply replacing fluorophore tags with stable isotopes of the heavy metals, and measuring these cell-by-cell with ICP-MS, dramatically increases the number of probes that can be simultaneously measured in cytometry and enables a transformative increase in the resolution of rare cell populations in complex biological samples. While this can be thought of as a novel incarnation of single-cell targeted proteomics, the metal-labeling reagents, ICP-MS of single cells, and accompanying informatics comprise a new field of technology termed Mass Cytometry. While the conception of mass cytometry is simple the embodiment to address the issues of multi-parameter flow cytometry has been far more challenging. There are many elements, and many more stable isotopes of those elements, that might be used as distinct reporter tags. Still, there are many approaches to conjugating metals to antibodies (or other affinity reagents) and work in this area along with developing new applications is ongoing. The mass resolution and linear (quantitative) dynamic range of ICP-MS allows those many stable isotopes to be measured simultaneously and without the spectral overlap issues that limit fluorescence assay. However, the adaptation of ICP-MS to allow high-speed simultaneous measurement with single cell distinction at high throughput required innovation of the cell introduction system, ion optics (sampling, transmission and beam-shaping), mass analysis, and signal handling and processing. An overview of “the nuts and bolts” of Mass Cytometry is presented.

Site-Specific, Covalent Immobilization of Dehalogenase ST2570 Catalyzed by Formylglycine-Generating Enzymes and Its Application in Batch and Semi-Continuous Flow Reactors.

PubMed

Jian, Hui; Wang, Yingwu; Bai, Yan; Li, Rong; Gao, Renjun

2016-07-11

Formylglycine-generating enzymes can selectively recognize and oxidize cysteine residues within the sulfatase sub motif at the terminus of proteins to form aldehyde-bearing formylglycine (FGly) residues, and are normally used in protein labeling. In this study, an aldehyde tag was introduced to proteins using formylglycine-generating enzymes encoded by a reconstructed set of the pET28a plasmid system for enzyme immobilization. The haloacid dehalogenase ST2570 from Sulfolobus tokodaii was used as a model enzyme. The C-terminal aldehyde-tagged ST2570 (ST2570CQ) exhibited significant enzymological properties, such as new free aldehyde groups, a high level of protein expression and improved enzyme activity. SBA-15 has widely been used as an immobilization support for its large surface and excellent thermal and chemical stability. It was functionalized with amino groups by aminopropyltriethoxysilane. The C-terminal aldehyde-tagged ST2570 was immobilized to SBA-15 by covalent binding. The site-specific immobilization of ST2570 avoided the chemical denaturation that occurs in general covalent immobilization and resulted in better fastening compared to physical adsorption. The site-specific immobilized ST2570 showed 3-fold higher thermal stability, 1.2-fold higher catalytic ability and improved operational stability than free ST2570. The site-specific immobilized ST2570 retained 60% of its original activity after seven cycles of batch operation, and it was superior to the ST2570 immobilized to SBA-15 by physical adsorption, which loses 40% of its original activity when used for the second time. It is remarkable that the site-specific immobilized ST2570 still retained 100% of its original activity after 10 cycles of reuse in the semi-continuous flow reactor. Overall, these results provide support for the industrial-scale production and application of site-specific, covalently immobilized ST2570.