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Sample records for specific activity measurement

  1. Evaluating bacterial activity from cell-specific ribosomal RNA content measured with oligonucleotide probes

    SciTech Connect

    Kemp, P.F.; Lee, S.; LaRoche, J.

    1992-10-01

    We describe a procedure for measuring the cell-specific quantity of ribosomal RNA (rRNA) and DNA in order to evaluate the frequency distribution of activity among cells. The procedure is inherently quantitative, does not require sample incubation and potentially can be taxon-specific. Fluorescently-labelled oligonucleotide probes are hybridized to the complementary 16S rRNA sequences in preserved, intact cells. The resulting cell fluorescence is proportional to cellular rRNA content and can be measured with a microscope-mounted photometer system, by image analysis, or by flow cytometry. Similarly, DNA content is measured as fluorescence of cells stained with the DNA specific fluorochrome DAPI. These are either prepared as separate samples for purposes of enumeration and DNA measurements, or are dual-labelled cells which are also hybridized with oligonucleotide probes.

  2. Evaluating bacterial activity from cell-specific ribosomal RNA content measured with oligonucleotide probes

    SciTech Connect

    Kemp, P.F.; Lee, S.; LaRoche, J.

    1992-01-01

    We describe a procedure for measuring the cell-specific quantity of ribosomal RNA (rRNA) and DNA in order to evaluate the frequency distribution of activity among cells. The procedure is inherently quantitative, does not require sample incubation and potentially can be taxon-specific. Fluorescently-labelled oligonucleotide probes are hybridized to the complementary 16S rRNA sequences in preserved, intact cells. The resulting cell fluorescence is proportional to cellular rRNA content and can be measured with a microscope-mounted photometer system, by image analysis, or by flow cytometry. Similarly, DNA content is measured as fluorescence of cells stained with the DNA specific fluorochrome DAPI. These are either prepared as separate samples for purposes of enumeration and DNA measurements, or are dual-labelled cells which are also hybridized with oligonucleotide probes.

  3. Specific activity measurement of ⁶⁴Cu: a comparison of methods.

    PubMed

    Mastren, Tara; Guthrie, James; Eisenbeis, Paul; Voller, Tom; Mebrahtu, Efrem; Robertson, J David; Lapi, Suzanne E

    2014-08-01

    Effective specific activity of (64)Cu (amount of radioactivity per µmol metal) is important in order to determine purity of a particular (64)Cu lot and to assist in optimization of the purification process. Metal impurities can affect effective specific activity and therefore it is important to have a simple method that can measure trace amounts of metals. This work shows that ion chromatography (IC) yields similar results to ICP mass spectrometry for copper, nickel and iron contaminants in (64)Cu production solutions. PMID:24709610

  4. Measurement of General and Specific Approaches to Physical Activity Parenting: A Systematic Review

    PubMed Central

    McDonald, Samantha; Cohen, Alysia

    2013-01-01

    Abstract Background Parents play a significant role in shaping youth physical activity (PA). However, interventions targeting PA parenting have been ineffective. Methodological inconsistencies related to the measurement of parental influences may be a contributing factor. The purpose of this article is to review the extant peer-reviewed literature related to the measurement of general and specific parental influences on youth PA. Methods A systematic review of studies measuring constructs of PA parenting was conducted. Computerized searches were completed using PubMed, MEDLINE, Academic Search Premier, SPORTDiscus, and PsycINFO. Reference lists of the identified articles were manually reviewed as well as the authors' personal collections. Articles were selected on the basis of strict inclusion criteria and details regarding the measurement protocols were extracted. A total of 117 articles met the inclusionary criteria. Methodological articles that evaluated the validity and reliability of PA parenting measures (n=10) were reviewed separately from parental influence articles (n=107). Results A significant percentage of studies used measures with indeterminate validity and reliability. A significant percentage of articles did not provide sample items, describe the response format, or report the possible range of scores. No studies were located that evaluated sensitivity to change. Conclusion The reporting of measurement properties and the use of valid and reliable measurement scales need to be improved considerably. PMID:23944923

  5. The roadmap for estimation of cell-type-specific neuronal activity from non-invasive measurements.

    PubMed

    Uhlirova, Hana; Kılıç, Kıvılcım; Tian, Peifang; Sakadžić, Sava; Gagnon, Louis; Thunemann, Martin; Desjardins, Michèle; Saisan, Payam A; Nizar, Krystal; Yaseen, Mohammad A; Hagler, Donald J; Vandenberghe, Matthieu; Djurovic, Srdjan; Andreassen, Ole A; Silva, Gabriel A; Masliah, Eliezer; Kleinfeld, David; Vinogradov, Sergei; Buxton, Richard B; Einevoll, Gaute T; Boas, David A; Dale, Anders M; Devor, Anna

    2016-10-01

    The computational properties of the human brain arise from an intricate interplay between billions of neurons connected in complex networks. However, our ability to study these networks in healthy human brain is limited by the necessity to use non-invasive technologies. This is in contrast to animal models where a rich, detailed view of cellular-level brain function with cell-type-specific molecular identity has become available due to recent advances in microscopic optical imaging and genetics. Thus, a central challenge facing neuroscience today is leveraging these mechanistic insights from animal studies to accurately draw physiological inferences from non-invasive signals in humans. On the essential path towards this goal is the development of a detailed 'bottom-up' forward model bridging neuronal activity at the level of cell-type-specific populations to non-invasive imaging signals. The general idea is that specific neuronal cell types have identifiable signatures in the way they drive changes in cerebral blood flow, cerebral metabolic rate of O2 (measurable with quantitative functional Magnetic Resonance Imaging), and electrical currents/potentials (measurable with magneto/electroencephalography). This forward model would then provide the 'ground truth' for the development of new tools for tackling the inverse problem-estimation of neuronal activity from multimodal non-invasive imaging data.This article is part of the themed issue 'Interpreting BOLD: a dialogue between cognitive and cellular neuroscience'. PMID:27574309

  6. Deep brain optical measurements of cell type–specific neural activity in behaving mice

    PubMed Central

    Cui, Guohong; Jun, Sang Beom; Jin, Xin; Luo, Guoxiang; Pham, Michael D; Lovinger, David M; Vogel, Steven S; Costa, Rui M

    2014-01-01

    Recent advances in genetically encoded fluorescent sensors enable the monitoring of cellular events from genetically defined groups of neurons in vivo. In this protocol, we describe how to use a time-correlated single-photon counting (tcspc)–based fiber optics system to measure the intensity, emission spectra and lifetime of fluorescent biosensors expressed in deep brain structures in freely moving mice. When combined with cre-dependent selective expression of genetically encoded ca2+ indicators (GecIs), this system can be used to measure the average neural activity from a specific population of cells in mice performing complex behavioral tasks. as an example, we used viral expression of GcaMps in striatal projection neurons (spns) and recorded the fluorescence changes associated with calcium spikes from mice performing a lever-pressing operant task. the whole procedure, consisting of virus injection, behavior training and optical recording, takes 3–4 weeks to complete. With minor adaptations, this protocol can also be applied to recording cellular events from other cell types in deep brain regions, such as dopaminergic neurons in the ventral tegmental area. the simultaneously recorded fluorescence signals and behavior events can be used to explore the relationship between the neural activity of specific brain circuits and behavior. PMID:24784819

  7. Mitochondrial Complex 1 Activity Measured by Spectrophotometry Is Reduced across All Brain Regions in Ageing and More Specifically in Neurodegeneration

    PubMed Central

    Chakrabarti, Lisa

    2016-01-01

    Mitochondrial function, in particular complex 1 of the electron transport chain (ETC), has been shown to decrease during normal ageing and in neurodegenerative disease. However, there is some debate concerning which area of the brain has the greatest complex 1 activity. It is important to identify the pattern of activity in order to be able to gauge the effect of age or disease related changes. We determined complex 1 activity spectrophotometrically in the cortex, brainstem and cerebellum of middle aged mice (70–71 weeks), a cerebellar ataxic neurodegeneration model (pcd5J) and young wild type controls. We share our updated protocol on the measurements of complex1 activity and find that mitochondrial fractions isolated from frozen tissues can be measured for robust activity. We show that complex 1 activity is clearly highest in the cortex when compared with brainstem and cerebellum (p<0.003). Cerebellum and brainstem mitochondria exhibit similar levels of complex 1 activity in wild type brains. In the aged brain we see similar levels of complex 1 activity in all three-brain regions. The specific activity of complex 1 measured in the aged cortex is significantly decreased when compared with controls (p<0.0001). Both the cerebellum and brainstem mitochondria also show significantly reduced activity with ageing (p<0.05). The mouse model of ataxia predictably has a lower complex 1 activity in the cerebellum, and although reductions are measured in the cortex and brain stem, the remaining activity is higher than in the aged brains. We present clear evidence that complex 1 activity decreases across the brain with age and much more specifically in the cerebellum of the pcd5j mouse. Mitochondrial impairment can be a region specific phenomenon in disease, but in ageing appears to affect the entire brain, abolishing the pattern of higher activity in cortical regions. PMID:27333203

  8. Comparison of measured and calculated concrete and rebar specific activity during decommissioning of the Dalhousie SLOWPOKE-2 reactor.

    PubMed

    Smith, Philip G; Everall, Dave; Ariani, Imelda; Tsang, Kwok

    2013-07-01

    Following the defuelling and dismantling of the Dalhousie University SLOWPOKE-2 Reactor (DUSR) in 2011, the reactor pool concrete and rebar were sampled to support the unconditional free release of the material such that the facility could be classified for unrestricted use. A detailed MCNP5 model of the critical core assembly was simulated to calculate the thermal, intermediate and fast neutron flux profile below the reactor pool floor. The neutron fluxes were used to calculate the specific activity of significant radionuclides in the concrete and rebar. The calculated specific activity and consequently the calculated neutron fluxes were validated at a number of sample locations. The calculated concrete and rebar specific activity were found to be in good agreement with the measured specific activity at the sample locations. The unrestricted use of the facility was granted through the approval of the licence to abandon the facility in August 2011. PMID:23658212

  9. Ultrasensitive and specific measurement of protease activity using functionalized photonic crystals.

    PubMed

    Gupta, Bakul; Mai, Kelly; Lowe, Stuart B; Wakefield, Denis; Di Girolamo, Nick; Gaus, Katharina; Reece, Peter J; Gooding, J Justin

    2015-10-01

    Herein is presented a microsensor technology as a diagnostic tool for detecting specific matrix metalloproteinases (MMPs) at very low concentrations. MMP-2 and MMP-9 are detected using label free porous silicon (PSi) photonic crystals that have been made selective for a given MMP by filling the nanopores with synthetic polymeric substrates containing a peptide sequence for that MMP. Proteolytic cleavage of the peptide sequence results in a shift in wavelength of the main peak in the reflectivity spectrum of the PSi device, which is dependent on the amount of MMP present. The ability to detect picogram amounts of MMP-2 and MMP-9 released by primary retinal pigment epithelial (RPE) cells and iris pigment epithelial (IPE) cells stimulated with lipopolysaccharide (LPS) is demonstrated. It was found that both cell types secrete higher amounts of MMP-2 than MMP-9 in their stimulated state, with RPE cells producing higher amounts of MMPs than IPE cells. The microsensor performance was compared to conventional protease detection systems, including gelatin zymography and enzyme linked immunosorbent assay (ELISA). It was found that the PSi microsensors were more sensitive than gelatin zymography; PSi microsensors detected the presence of both MMP-2 and MMP-9 while zymography could only detect MMP-2. The MMP-2 and MMP-9 quantification correlated well with the ELISA. This new method of detecting protease activity shows superior performance to conventional protease assays and has the potential for translation to high-throughput multiplexed analysis. PMID:26312479

  10. Laboratory Measurement of the Anticoagulant Activity of the Target-specific Oral Anticoagulant Agents: A Systematic Review

    PubMed Central

    Cuker, Adam; Siegal, Deborah M.; Crowther, Mark A.; Garcia, David A.

    2014-01-01

    Background The target-specific oral anticoagulant agents (TSOACs) do not require routine laboratory monitoring. However, laboratory measurement may be desirable in special situations and populations. Objectives This study’s objective is to systematically review and summarize current evidence regarding laboratory measurement of the anticoagulant activity of dabigatran, rivaroxaban, and apixaban. Methods We searched PubMed and Web of Science for studies that reported a relationship between drug levels of dabigatran, rivaroxaban, and apixaban and coagulation assay results. Study quality was evaluated using Quality Assessment of Diagnostic Accuracy Studies-2 (QUADAS-2). Results We identified 17 eligible studies for dabigatran, 15 for rivaroxaban, and 4 for apixaban. For dabigatran, a normal thrombin time excludes clinically relevant drug concentrations. The activated partial thromboplastin time (APTT) and prothrombin time (PT) are less sensitive and may be normal at trough drug levels. The dilute thrombin time (R2 0.92–0.99) and ecarin-based assays (R2 0.92–1.00) show excellent linearity across on-therapy drug concentrations and may be used for drug quantification. In terms of rivaroxaban and apixaban, anti-Xa activity is linear (R2 0.89–1.00) over a wide range of drug levels and may be used for drug quantification. Undetectable anti-Xa activity likely excludes clinically relevant drug concentrations. The PT is less sensitive (especially for apixaban); a normal PT may not exclude clinically relevant levels. The APTT demonstrates insufficient sensitivity and linearity for quantification. Conclusions Dabigatran, rivaroxaban, and apixaban exhibit variable effects on coagulation assays. Understanding these effects facilitates interpretation of test results in TSOAC-treated patients. More information on the relationship between drug levels and clinical outcomes is needed. PMID:25212648

  11. Measurement of transformation temperatures and specific heat capacity of tungsten added reduced activation ferritic-martensitic steel

    NASA Astrophysics Data System (ADS)

    Raju, S.; Jeya Ganesh, B.; Rai, Arun Kumar; Mythili, R.; Saroja, S.; Mohandas, E.; Vijayalakshmi, M.; Rao, K. B. S.; Raj, Baldev

    2009-06-01

    The on-heating phase transformation temperatures up to the melting regime and the specific heat capacity of a reduced activation ferritic-martensitic steel (RAFM) with a nominal composition (wt%): 9Cr-0.09C-0.56Mn-0.23V-1W-0.063Ta-0.02N, have been measured using high temperature differential scanning calorimetry. The α -ferrite + carbides → γ-austenite transformation start and finish temperatures, namely A c1, and A c3, are found to be 1104 and 1144 K, respectively for a typical normalized and tempered microstructure. It is also observed that the martensite start ( MS) and finish ( Mf) temperatures are sensitive to the austenitising conditions. Typical MS and Mf values for the 1273 K normalized and 1033 K tempered samples are of the order 714 and 614 K, respectively. The heat capacity CP of the RAFM steel has been measured in the temperature range 473-1273 K, for different normalized and tempered samples. In essence, it is found that the CP of the fully martensitic microstructure is found to be lower than that of its tempered counterpart, and this difference begins to increase in an appreciable manner from about 800 K. The heat capacity of the normalized microstructure is found to vary from 480 to 500 J kg -1 K -1 at 500 K, where as that of the tempered steel is found to be higher by about, 150 J kg -1 K -1.

  12. An update on laboratory measurements of Dabigatran: Smart specific and calibrated dedicated assays for measuring anti-IIa activity in plasma.

    PubMed

    Amiral, Jean; Dunois, Claire; Amiral, Cédric; Seghatchian, Jerard

    2016-06-01

    Use of Direct Oral Anticoagulants (DOACs) is continuously increasing for clinical application. The first product released was Dabigatran, which was proposed for many preventive and curative applications, especially for prevention of stroke in patients with non-valvular atrial fibrillation. Although measurement of Dabigatran Anti-IIa activity in plasma is not requested on a routine basis, in some situations its measurement is clinically useful. Especially, before an emergency surgery in treated patients, where its presence at high concentrations, which will expose the patient at an increased bleeding risk, has to be excluded. Hence, smart, specific, rapid and accurate quantitative assays are warranted as an essential required. Hemoclot™ Thrombin Inhibitors and Biophen® DTI were specifically designed for these applications, and can be used on all automated instruments with a standard range protocol for measuring concentrations at peak, or with a low range protocol for testing residual concentrations. Both functional assays have a good correlation with the reference LC-MS/MS method, and concentrations measured are similar. Performances of these assays and interferences of various substances or drugs are discussed. Some differences in variations of clotting times are observed between mechanical or optical clot detection instruments, which could be explained by the fibrin clot structure, altered by direct Factor Xa inhibitors, and more especially Rivaroxaban. Both clotting and chromogenic assays offer a safe and accurate quantitative measurement of Dabigatran in plasma in all situations where this determination is requested. In short this manuscript provides an in depth update on current opinions on laboratory aspects of measuring Dabigatran concentrations in plasma, when required. PMID:27216543

  13. Total cyanide mass measurement with micro-ion selective electrode for determination of specific activity of carbon-11 cyanide

    DOE PAGESBeta

    Shea, Colleen; Alexoff, David L.; Kim, Dohyun; Hoque, Ruma; Schueller, Michael J.; Fowler, Joanna S.; Qu, Wenchao

    2015-04-25

    In this study, we aim to directly measure the specific activity (SA) of the carbon-11 cyanide ([11C]CN¯) produced by our in-house built automated [11C]HCN production system and to identify the major sources of 12C-cyanide (12CN¯). The [11C]CN¯ is produced from [11C]CO2, which is generated by the 14N(p,α)11C nuclear reaction using a cyclotron. Direct measurement of cyanide concentrations was accomplished using a relatively inexpensive, and easy to use ion selective electrode (ISE) which offered an appropriate range of sensitivity for detecting mass. Multiple components of the [11C]HCN production system were isolated in order to determine their relative contributions to 12CN¯ mass.more » It was determined that the system gases were responsible for approximately 30% of the mass, and that the molecular sieve/nickel furnace unit contributed approximately 70% of the mass. Beam on target (33 µA for 1 and 10 min) did not contribute significantly to the mass. Additionally, we compared the SA of our [11C]HCN precursor determined using the ISE to the SA of our current [11C]CN¯ derived radiotracers determined by HPLC to assure there was no significant difference between the two methods. These results are the first reported use of an ion selective electrode to determine the SA of no-carrier-added cyanide ion, and clearly show that it is a valuable, inexpensive and readily available tool suitable for this purpose.« less

  14. Total cyanide mass measurement with micro-ion selective electrode for determination of specific activity of carbon-11 cyanide

    SciTech Connect

    Shea, Colleen; Alexoff, David L.; Kim, Dohyun; Hoque, Ruma; Schueller, Michael J.; Fowler, Joanna S.; Qu, Wenchao

    2015-04-25

    In this study, we aim to directly measure the specific activity (SA) of the carbon-11 cyanide ([11C]CN¯) produced by our in-house built automated [11C]HCN production system and to identify the major sources of 12C-cyanide (12CN¯). The [11C]CN¯ is produced from [11C]CO2, which is generated by the 14N(p,α)11C nuclear reaction using a cyclotron. Direct measurement of cyanide concentrations was accomplished using a relatively inexpensive, and easy to use ion selective electrode (ISE) which offered an appropriate range of sensitivity for detecting mass. Multiple components of the [11C]HCN production system were isolated in order to determine their relative contributions to 12CN¯ mass. It was determined that the system gases were responsible for approximately 30% of the mass, and that the molecular sieve/nickel furnace unit contributed approximately 70% of the mass. Beam on target (33 µA for 1 and 10 min) did not contribute significantly to the mass. Additionally, we compared the SA of our [11C]HCN precursor determined using the ISE to the SA of our current [11C]CN¯ derived radiotracers determined by HPLC to assure there was no significant difference between the two methods. These results are the first reported use of an ion selective electrode to determine the SA of no-carrier-added cyanide ion, and clearly show that it is a valuable, inexpensive and readily available tool suitable for this purpose.

  15. Frequency specific interactions of MEG resting state activity within and across brain networks as revealed by the multivariate interaction measure.

    PubMed

    Marzetti, L; Della Penna, S; Snyder, A Z; Pizzella, V; Nolte, G; de Pasquale, F; Romani, G L; Corbetta, M

    2013-10-01

    Resting state networks (RSNs) are sets of brain regions exhibiting temporally coherent activity fluctuations in the absence of imposed task structure. RSNs have been extensively studied with fMRI in the infra-slow frequency range (nominally <10(-1)Hz). The topography of fMRI RSNs reflects stationary temporal correlation over minutes. However, neuronal communication occurs on a much faster time scale, at frequencies nominally in the range of 10(0)-10(2)Hz. We examined phase-shifted interactions in the delta (2-3.5 Hz), theta (4-7 Hz), alpha (8-12 Hz) and beta (13-30 Hz) frequency bands of resting-state source space MEG signals. These analyses were conducted between nodes of the dorsal attention network (DAN), one of the most robust RSNs, and between the DAN and other networks. Phase shifted interactions were mapped by the multivariate interaction measure (MIM), a measure of true interaction constructed from the maximization of imaginary coherency in the virtual channels comprised of voxel signals in source space. Non-zero-phase interactions occurred between homologous left and right hemisphere regions of the DAN in the delta and alpha frequency bands. Even stronger non-zero-phase interactions were detected between networks. Visual regions bilaterally showed phase-shifted interactions in the alpha band with regions of the DAN. Bilateral somatomotor regions interacted with DAN nodes in the beta band. These results demonstrate the existence of consistent, frequency specific phase-shifted interactions on a millisecond time scale between cortical regions within RSN as well as across RSNs. PMID:23631996

  16. Effect of heavy metals on nitrification activity as measured by RNA- and DNA-based function-specific assays

    EPA Science Inventory

    Heavy metals can inhibit nitrification, a key process for nitrogen removal in wastewater treatment. The transcriptional responses of functional genes (amoA, hao, nirK and norB) were measured in conjunction with specific oxygen uptake rate (sOUR) for nitrifying enrichment cultures...

  17. Measurement of HCV-Specific CD8(+) Cytotoxic T-Cell Activities in the Peripheral Blood by Europium Release Assay.

    PubMed

    Imawari, M

    1999-01-01

    Peripheral blood mononuclear cells (PBMC) contain NK cells, cytotoxic T-lymphocytes (CTL), helper T-cells, and B-cells that respond to viral infection and act to eliminate the virus from infected individuals. CTLs are not only thought to be a major host defense against viral infection, but are also implicated in the immunopathogenesis. Classical CTLs are CD8(+) and recognize endogenously synthesized and processed antigen in association with a human leukocyte antigen (HLA) class I molecule. The antigens are usually 8-10 amino acids long. HCV-specific CTLs have been demonstrated in the peripheral blood of some of patients with HCV infection by stimulating PBMC with the HCV synthetic peptides (1). The peptides were synthesized as overlapping peptides to encompass a certain region of the HCV antigen (1), on the basis of antigenicity prediction from the amino acid composition of HCV (2), or on the basis of the HLA binding motifs in the HCV antigen (3). Several minimal and optimal epitopes in the HCV antigen and their HLA restriction of recognition by CTLs have been defined. Recently, it has been reported that HCV-specific CTLs may suppress the outgrowth of HCV (4). In this chapter, methods will be discussed that demonstrate HCV-specific CTLs in the peripheral blood of patients with HCV infection. We use nonradioisotope europium (Eu) for assay of CTL activities. PMID:21374383

  18. Measuring Physical Activity Environments

    PubMed Central

    Sallis, James F.

    2010-01-01

    Physical activity is usually done in specific types of places, referred to as physical activity environments. These often include parks, trails, fitness centers, schools, and streets. In recent years, scientific interest has increased notably in measuring physical activity environments. The present paper provides an historical overview of the contributions of the health, planning, and leisure studies fields to the development of contemporary measures. The emphasis is on attributes of the built environment that can be affected by policies to contribute to the promotion of physical activity. Researchers from health fields assessed a wide variety of built environment variables expected to be related to recreational physical activity. Settings of interest were schools, workplaces, and recreation facilities, and most early measures used direct observation methods with demonstrated inter-observer reliability. Investigators from the city planning field evaluated aspects of community design expected to be related to people’s ability to walk from homes to destinations. GIS was used to assess walkability defined by the 3Ds of residential density, land-use diversity, and pedestrian-oriented designs. Evaluating measures for reliability or validity was rarely done in the planning-related fields. Researchers in the leisure studies and recreation fields studied mainly people’s use of leisure time rather than physical characteristics of parks and other recreation facilities. Although few measures of physical activity environments were developed, measures of aesthetic qualities are available. Each of these fields made unique contributions to the contemporary methods used to assess physical activity environments. PMID:19285214

  19. Analysing specificity of a bipolar EEG measurement.

    PubMed

    Vaisanen, Juho; Ryynanen, Outi; Hyttinen, Jari; Malmivuo, Jaakko

    2006-01-01

    The objective in bioelectric measurements such as ECG and EEG is to register the signal arising from sources in the region of interest. It is also desired that signal-to-noise ratio (SNR) of a measurement is high. The sensitivity of an ideal measurement should focus on and be greater on the target areas in comparison to other areas of the volume conductor. Previously the half-sensitivity volume (HSV) has been applied to describe how focused the measurement is. In this paper we introduce a concept of the half-sensitivity ratio (HSR) which describes how well the sensitivity is concentrated in HSV compared to other source regions i.e. how specific the measurement is to the sources in HSV. Further we may have different region of interests (ROI) to which the measurements are wanted to be specific. Then the concept is called region of interest sensitivity ratio (ROISR). We present here an application of the HSR in analysing sensitivity distributions of bioelectric measurements. We studied the effects of interelectrode distance and the scalp/skull/brain resistivity ratio on the HSR of a bipolar EEG measurement with a three-layer spherical head model. The results indicate that when the focus of interest is on cortical activity more specified and concentrated sensitivity distributions are achieved with smaller interelectrode distances. Further a preliminary measurement with visual evoked potentials provides evidence of the relationship between HSR and SNR of a measurement. PMID:17945619

  20. Measurement of lysine-specific demethylase-1 activity in the nuclear extracts by flow-injection based time-of-flight mass spectrometry

    PubMed Central

    Sakane, Chiharu; Ohta, Hiromichi; Shidoji, Yoshihiro

    2015-01-01

    Lysine-specific demethylase 1 (LSD1/KDM1A), a histone-modifying enzyme, is upregulated in many cancers, especially in neuroblastoma, breast cancer and hepatoma. We have established a simple method to measure LSD1 activity using a synthetic N-terminal 21-mer peptide of histone H3, which is dimethylated at Lys-4 (H3K4me2). After the enzyme reaction, a substrate of H3K4me2 and two demethylated products, H3K4me1 and H3K4me0, were quantitatively determined by flow injection time-of-flight mass spectrometry (FI-TOF/MS). By using recombinant human LSD1, a nonlinear fitting simulation of the data obtained by FI-TOF/MS produced typical consecutive-reaction kinetics. Apparent Km and kcat values of hLSD1 for the first and second demethylation reactions were found to be in the range of reported values. Tranylcypromine was shown to inhibit LSD1 activity with an IC50 of 6.9 µM for the first demethylation reaction and 5.8 µM for the second demethylation reaction. The FI-TOF/MS assay revealed that the endogenous LSD1 activity was higher in the nuclear extracts of SH-SY5Y cells than in HeLa or PC-3 cells, and this is in accordance with the immunoblotting data using an anti-LSD1 antibody. A simple, straightforward FI-TOF/MS assay is described to efficiently measure LSD1 activity in the nuclear extracts of cultured cells. PMID:25759518

  1. Measurement of lysine-specific demethylase-1 activity in the nuclear extracts by flow-injection based time-of-flight mass spectrometry.

    PubMed

    Sakane, Chiharu; Ohta, Hiromichi; Shidoji, Yoshihiro

    2015-03-01

    Lysine-specific demethylase 1 (LSD1/KDM1A), a histone-modifying enzyme, is upregulated in many cancers, especially in neuroblastoma, breast cancer and hepatoma. We have established a simple method to measure LSD1 activity using a synthetic N-terminal 21-mer peptide of histone H3, which is dimethylated at Lys-4 (H3K4me2). After the enzyme reaction, a substrate of H3K4me2 and two demethylated products, H3K4me1 and H3K4me0, were quantitatively determined by flow injection time-of-flight mass spectrometry (FI-TOF/MS). By using recombinant human LSD1, a nonlinear fitting simulation of the data obtained by FI-TOF/MS produced typical consecutive-reaction kinetics. Apparent K m and k cat values of hLSD1 for the first and second demethylation reactions were found to be in the range of reported values. Tranylcypromine was shown to inhibit LSD1 activity with an IC50 of 6.9 µM for the first demethylation reaction and 5.8 µM for the second demethylation reaction. The FI-TOF/MS assay revealed that the endogenous LSD1 activity was higher in the nuclear extracts of SH-SY5Y cells than in HeLa or PC-3 cells, and this is in accordance with the immunoblotting data using an anti-LSD1 antibody. A simple, straightforward FI-TOF/MS assay is described to efficiently measure LSD1 activity in the nuclear extracts of cultured cells. PMID:25759518

  2. Measurement of Physical Activity.

    ERIC Educational Resources Information Center

    Dishman, Rod K.; Washburn, Richard A.; Schoeller, Dale A.

    2001-01-01

    Valid assessment of physical activity must be unobtrusive, practical to administer, and specific about physical activity type, frequency, duration, and intensity. Assessment methods can be categorized according to whether they provide direct or indirect (e.g., self-report) observation of physical activity, body motion, physiological response…

  3. High specific activity silicon-32

    DOEpatents

    Phillips, D.R.; Brzezinski, M.A.

    1996-06-11

    A process for preparation of silicon-32 is provided and includes contacting an irradiated potassium chloride target, including spallation products from a prior irradiation, with sufficient water, hydrochloric acid or potassium hydroxide to form a solution, filtering the solution, adjusting pH of the solution from about 5.5 to about 7.5, admixing sufficient molybdate-reagent to the solution to adjust the pH of the solution to about 1.5 and to form a silicon-molybdate complex, contacting the solution including the silicon-molybdate complex with a dextran-based material, washing the dextran-based material to remove residual contaminants such as sodium-22, separating the silicon-molybdate complex from the dextran-based material as another solution, adding sufficient hydrochloric acid and hydrogen peroxide to the solution to prevent reformation of the silicon-molybdate complex and to yield an oxidation state of the molybdate adapted for subsequent separation by an anion exchange material, contacting the solution with an anion exchange material whereby the molybdate is retained by the anion exchange material and the silicon remains in solution, and optionally adding sufficient alkali metal hydroxide to adjust the pH of the solution to about 12 to 13. Additionally, a high specific activity silicon-32 product having a high purity is provided.

  4. High specific activity silicon-32

    DOEpatents

    Phillips, Dennis R.; Brzezinski, Mark A.

    1996-01-01

    A process for preparation of silicon-32 is provided and includes contacting an irradiated potassium chloride target, including spallation products from a prior irradiation, with sufficient water, hydrochloric acid or potassium hydroxide to form a solution, filtering the solution, adjusting pH of the solution to from about 5.5 to about 7.5, admixing sufficient molybdate-reagent to the solution to adjust the pH of the solution to about 1.5 and to form a silicon-molybdate complex, contacting the solution including the silicon-molybdate complex with a dextran-based material, washing the dextran-based material to remove residual contaminants such as sodium-22, separating the silicon-molybdate complex from the dextran-based material as another solution, adding sufficient hydrochloric acid and hydrogen peroxide to the solution to prevent reformation of the silicon-molybdate complex and to yield an oxidization state of the molybdate adapted for subsequent separation by an anion exchange material, contacting the solution with an anion exchange material whereby the molybdate is retained by the anion exchange material and the silicon remains in solution, and optionally adding sufficient alkali metal hydroxide to adjust the pH of the solution to about 12 to 13. Additionally, a high specific activity silicon-32 product having a high purity is provided.

  5. Production of high specific activity silicon-32

    SciTech Connect

    Phillips, D.R.; Brzezinski, M.A.

    1998-12-31

    This is the final report of a three-year, Laboratory Directed Research and Development Project (LDRD) at Los Alamos National Laboratory (LANL). There were two primary objectives for the work performed under this project. The first was to take advantage of capabilities and facilities at Los Alamos to produce the radionuclide {sup 32}Si in unusually high specific activity. The second was to combine the radioanalytical expertise at Los Alamos with the expertise at the University of California to develop methods for the application of {sup 32}Si in biological oceanographic research related to global climate modeling. The first objective was met by developing targetry for proton spallation production of {sup 32}Si in KCl targets and chemistry for its recovery in very high specific activity. The second objective was met by developing a validated field-useable, radioanalytical technique, based upon gas-flow proportional counting, to measure the dynamics of silicon uptake by naturally occurring diatoms.

  6. Disease Activity Measures in Paediatric Rheumatic Diseases

    PubMed Central

    Luca, Nadia J.; Feldman, Brian M.

    2013-01-01

    Disease activity refers to potentially reversible aspects of a disease. Measurement of disease activity in paediatric rheumatic diseases is a critical component of patient care and clinical research. Disease activity measures are developed systematically, often involving consensus methods. To be useful, a disease activity measure must be feasible, valid, and interpretable. There are several challenges in quantifying disease activity in paediatric rheumatology; namely, the conditions are multidimensional, the level of activity must be valuated in the context of treatment being received, there is no gold standard for disease activity, and it is often difficult to incorporate the patient's perspective of their disease activity. To date, core sets of response variables are defined for juvenile idiopathic arthritis, juvenile systemic lupus erythematosus, and juvenile dermatomyositis, as well as definitions for improvement in response to therapy. Several specific absolute disease activity measures also exist for each condition. Further work is required to determine the optimal disease activity measures in paediatric rheumatology. PMID:24089617

  7. MEASUREMENT OF SPECIFIC HEAT CAPACITY OF SALTSTONE

    SciTech Connect

    Harbour, J; Vickie Williams, V

    2008-09-29

    One of the goals of the Saltstone variability study is to identify (and quantify the impact of) the operational and compositional variables that control or influence the important processing and performance properties of Saltstone grout mixtures. The heat capacity of the Saltstone waste form is one of the important properties of Saltstone mixes that was last measured at SRNL in 1997. It is therefore important to develop a core competency for rapid and accurate analysis of the specific heat capacity of the Saltstone mixes in order to quantify the impact of compositional and operational variations on this property as part of the variability study. The heat capacity, coupled with the heat of hydration data obtained from isothermal calorimetry for a given Saltstone mix, can be used to predict the maximum temperature increase in the cells within the vaults of the Saltstone Disposal Facility (SDF). The temperature increase controls the processing rate and the pour schedule. The maximum temperature is also important to the performance properties of the Saltstone. For example, in mass pours of concrete or grout of which Saltstone is an example, the maximum temperature increase and the maximum temperature difference (between the surface and the hottest location) are controlled to ensure durability of the product and prevent or limit the cracking caused by the thermal gradients produced during curing. This report details the development and implementation of a method for the measurement of the heat capacities of Saltstone mixes as well as the heat capacities of the cementitious materials of the premix and the simulated salt solutions used to batch the mixes. The developed method utilizes the TAM Air isothermal calorimeter and takes advantage of the sophisticated heat flow measurement capabilities of the instrument. Standards and reference materials were identified and used to validate the procedure and ensure accuracy of testing. Heat capacities of Saltstone mixes were

  8. Enzyme Specific Activity in Functionalized Nanoporous Supports

    SciTech Connect

    Lei, Chenghong; Soares, Thereza A.; Shin, Yongsoon; Liu, Jun; Ackerman, Eric J.

    2008-03-26

    Enzyme specific activity can be increased or decreased to a large extent by changing protein loading density in functionalized nanoporous support, where organophosphorus hydrolase can display a constructive orientation and thus leave a completely open entrance for substrate even at higher protein loading density, but glucose oxidase can not.

  9. Measurement of phenotype and absolute number of circulating heparin-binding hemagglutinin, ESAT-6 and CFP-10, and purified protein derivative antigen-specific CD4 T cells can discriminate active from latent tuberculosis infection.

    PubMed

    Hutchinson, Paul; Barkham, Timothy M S; Tang, Wenying; Kemeny, David M; Chee, Cynthia Bin-Eng; Wang, Yee T

    2015-02-01

    The tuberculin skin test (TST) and interferon gamma (IFN-γ) release assays (IGRAs) are used as adjunctive tests for the evaluation of suspected cases of active tuberculosis (TB). However, a positive test does not differentiate latent from active TB. We investigated whether flow cytometric measurement of novel combinations of intracellular cytokines and surface makers on CD4 T cells could differentiate between active and latent TB after stimulation with Mycobacterium tuberculosis-specific proteins. Blood samples from 60 patients referred to the Singapore Tuberculosis Control Unit for evaluation for active TB or as TB contacts were stimulated with purified protein derivative (PPD), ESAT-6 and CFP-10, or heparin-binding hemagglutinin (HBHA). The CD4 T cell cytokine response (IFN-γ, interleukin-2 [IL-2], interleukin-17A [IL-17A], interleukin-22 [IL-22], granulocyte-macrophage colony-stimulating factor [GM-CSF], and tumor necrosis factor alpha [TNF-α]) and surface marker expression (CD27, CXCR3, and CD154) were then measured. We found that the proportion of PPD-specific CD4 T cells, defined as CD154(+) TNF-α(+) cells that were negative for CD27 and positive for GM-CSF, gave the strongest discrimination between subjects with latent and those with active TB (area under the receiver operator characteristic [ROC] curve of 0.9277; P < 0.0001). Also, the proportions and absolute numbers of HBHA-specific CD4 T cells were significantly higher in those with latent TB infection, particularly CD154(+) TNF-α(+) IFN-γ(+) IL-2(+) and CD154(+) TNF-α(+) CXCR3(+). Finally, we found that the ratio of ESAT-6- and CFP-10-responding to HBHA-responding CD4 T cells was significantly different between the two study populations. In conclusion, we found novel markers of M. tuberculosis-specific CD4 cells which differentiate between active and latent TB. PMID:25520147

  10. Measuring activity in ant colonies

    NASA Astrophysics Data System (ADS)

    Noda, C.; Fernández, J.; Pérez-Penichet, C.; Altshuler, E.

    2006-12-01

    Ants, as paradigm of social insects, have become a recurrent example of efficient problem solvers via self-organization. In spite of the simple behavior of each individual, the colony as a whole displays "swarm intelligence:" the organization of ant trails for foraging is a typical output of it. But conventional techniques of observation can hardly record the amount of data needed to get a detailed understanding of self-organization of ant swarms in the wild. Here we are presenting a measurement system intended to monitor ant activity in the field comprising massive data acquisition and high sensitivity. A central role is played by an infrared sensor devised specifically to monitor relevant parameters to the activity of ants through the exits of the nest, although other sensors detecting temperature and luminosity are added to the system. We study the characteristics of the activity sensor and its performance in the field. Finally, we present massive data measured at one exit of a nest of Atta insularis, an ant endemic to Cuba, to illustrate the potential of our system.

  11. Spectral measurements in critical assemblies: MCNP specifications and calculated results

    SciTech Connect

    Stephanie C. Frankle; Judith F. Briesmeister

    1999-12-01

    Recently, a suite of 86 criticality benchmarks for the Monte Carlo N-Particle (MCNP) transport code was developed, and the results of testing the ENDF/B-V and ENDF/B-VI data (through Release 2) were published. In addition to the standard k{sub eff} measurements, other experimental measurements were performed on a number of these benchmark assemblies. In particular, the Cross Section Evaluation Working Group (CSEWG) specifications contain experimental data for neutron leakage and central-flux measurements, central-fission ratio measurements, and activation ratio measurements. Additionally, there exists another set of fission reaction-rate measurements performed at the National Institute of Standards and Technology (NIST) utilizing a {sup 252}Cf source. This report will describe the leakage and central-flux measurements and show a comparison of experimental data to MCNP simulations performed using the ENDF/B-V and B-VI (Release 2) data libraries. Central-fission and activation reaction-rate measurements will be described, and the comparison of experimental data to MCNP simulations using available data libraries for each reaction of interest will be presented. Finally, the NIST fission reaction-rate measurements will be described. A comparison of MCNP results published previously with the current MCNP simulations will be presented for the NIST measurements, and a comparison of the current MCNP simulations to the experimental measurements will be presented.

  12. Active site specificity of plasmepsin II.

    PubMed Central

    Westling, J.; Cipullo, P.; Hung, S. H.; Saft, H.; Dame, J. B.; Dunn, B. M.

    1999-01-01

    Members of the aspartic proteinase family of enzymes have very similar three-dimensional structures and catalytic mechanisms. Each, however, has unique substrate specificity. These distinctions arise from variations in amino acid residues that line the active site subsites and interact with the side chains of the amino acids of the peptides that bind to the active site. To understand the unique binding preferences of plasmepsin II, an enzyme of the aspartic proteinase class from the malaria parasite, Plasmodium falciparum, chromogenic octapeptides having systematic substitutions at various positions in the sequence were analyzed. This enabled the design of new, improved substrates for this enzyme (Lys-Pro-Ile-Leu-Phe*Nph-Ala/Glu-Leu-Lys, where * indicates the cleavage point). Additionally, the crystal structure of plasmepsin II was analyzed to explain the binding characteristics. Specific amino acids (Met13, Ser77, and Ile287) that were suspected of contributing to active site binding and specificity were chosen for site-directed mutagenesis experiments. The Met13Glu and Ile287Glu single mutants and the Met13Glu/Ile287Glu double mutant gain the ability to cleave substrates containing Lys residues. PMID:10548045

  13. High specific activity platinum-195m

    DOEpatents

    Mirzadeh, Saed; Du, Miting; Beets, Arnold L.; Knapp, Jr., Furn F.

    2004-10-12

    A new composition of matter includes .sup.195m Pt characterized by a specific activity of at least 30 mCi/mg Pt, generally made by method that includes the steps of: exposing .sup.193 Ir to a flux of neutrons sufficient to convert a portion of the .sup.193 Ir to .sup.195m Pt to form an irradiated material; dissolving the irradiated material to form an intermediate solution comprising Ir and Pt; and separating the Pt from the Ir by cation exchange chromatography to produce .sup.195m Pt.

  14. Production Of High Specific Activity Copper-67

    DOEpatents

    Jamriska, Sr., David J.; Taylor, Wayne A.; Ott, Martin A.; Fowler, Malcolm; Heaton, Richard C.

    2003-10-28

    A process for the selective production and isolation of high specific activity Cu.sup.67 from proton-irradiated enriched Zn.sup.70 target comprises target fabrication, target irradiation with low energy (<25 MeV) protons, chemical separation of the Cu.sup.67 product from the target material and radioactive impurities of gallium, cobalt, iron, and stable aluminum via electrochemical methods or ion exchange using both anion and cation organic ion exchangers, chemical recovery of the enriched Zn.sup.70 target material, and fabrication of new targets for re-irradiation is disclosed.

  15. Production Of High Specific Activity Copper-67

    DOEpatents

    Jamriska, Sr., David J.; Taylor, Wayne A.; Ott, Martin A.; Fowler, Malcolm; Heaton, Richard C.

    2002-12-03

    A process for the selective production and isolation of high specific activity cu.sup.67 from proton-irradiated enriched Zn.sup.70 target comprises target fabrication, target irradiation with low energy (<25 MeV) protons, chemical separation of the Cu.sup.67 product from the target material and radioactive impurities of gallium, cobalt, iron, and stable aluminum via electrochemical methods or ion exchange using both anion and cation organic ion exchangers, chemical recovery of the enriched Zn.sup.70 target material, and fabrication of new targets for re-irradiation is disclosed.

  16. Measuring Language-Specific Phonetic Settings

    ERIC Educational Resources Information Center

    Mennen, Ineke; Scobbie, James M.; de Leeuw, Esther; Schaeffler, Sonja; Schaeffler, Felix

    2010-01-01

    While it is well known that languages have different phonemes and phonologies, there is growing interest in the idea that languages may also differ in their "phonetic setting". The term "phonetic setting" refers to a tendency to make the vocal apparatus employ a language-specific habitual configuration. For example, languages may differ in their…

  17. 40 CFR 89.415 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 21 2012-07-01 2012-07-01 false Fuel flow measurement specifications... Emission Test Procedures § 89.415 Fuel flow measurement specifications. The fuel flow rate measurement... parameters are the elapsed time measurement of the event and the weight or volume measurement....

  18. 40 CFR 89.415 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Fuel flow measurement specifications... Emission Test Procedures § 89.415 Fuel flow measurement specifications. The fuel flow rate measurement... parameters are the elapsed time measurement of the event and the weight or volume measurement....

  19. 40 CFR 86.314-79 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... percent of the measuring weight. (3) If the mass of fuel consumed is measured electronically (load cell... 40 Protection of Environment 18 2011-07-01 2011-07-01 false Fuel flow measurement specifications....314-79 Fuel flow measurement specifications. (a) The fuel flow rate measurement instrument must have...

  20. Mechanisms of Specificity for Hox Factor Activity

    PubMed Central

    Zandvakili, Arya; Gebelein, Brian

    2016-01-01

    Metazoans encode clusters of paralogous Hox genes that are critical for proper development of the body plan. However, there are a number of unresolved issues regarding how paralogous Hox factors achieve specificity to control distinct cell fates. First, how do Hox paralogs, which have very similar DNA binding preferences in vitro, drive different transcriptional programs in vivo? Second, the number of potential Hox binding sites within the genome is vast compared to the number of sites bound. Hence, what determines where in the genome Hox factors bind? Third, what determines whether a Hox factor will activate or repress a specific target gene? Here, we review the current evidence that is beginning to shed light onto these questions. In particular, we highlight how cooperative interactions with other transcription factors (especially PBC and HMP proteins) and the sequences of cis-regulatory modules provide a basis for the mechanisms of Hox specificity. We conclude by integrating a number of the concepts described throughout the review in a case study of a highly interrogated Drosophila cis-regulatory module named “The Distal-less Conserved Regulatory Element” (DCRE). PMID:27583210

  1. Functional Specificity of the Visual Word Form Area: General Activation for Words and Symbols but Specific Network Activation for Words

    ERIC Educational Resources Information Center

    Reinke, Karen; Fernandes, Myra; Schwindt, Graeme; O'Craven, Kathleen; Grady, Cheryl L.

    2008-01-01

    The functional specificity of the brain region known as the Visual Word Form Area (VWFA) was examined using fMRI. We explored whether this area serves a general role in processing symbolic stimuli, rather than being selective for the processing of words. Brain activity was measured during a visual 1-back task to English words, meaningful symbols…

  2. 40 CFR 91.417 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 21 2013-07-01 2013-07-01 false Fuel flow measurement specifications... Procedures § 91.417 Fuel flow measurement specifications. (a) Fuel flow measurement is required only for raw testing but is allowed for dilute testing. (b) The fuel flow rate measurement instrument must have...

  3. 40 CFR 91.417 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 20 2011-07-01 2011-07-01 false Fuel flow measurement specifications... Procedures § 91.417 Fuel flow measurement specifications. (a) Fuel flow measurement is required only for raw testing but is allowed for dilute testing. (b) The fuel flow rate measurement instrument must have...

  4. 40 CFR 86.314-79 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 18 2010-07-01 2010-07-01 false Fuel flow measurement specifications....314-79 Fuel flow measurement specifications. (a) The fuel flow rate measurement instrument must have a... period of the clock. (2) For Diesel engines only, if the mass of fuel consumed is measured by...

  5. 40 CFR 86.314-79 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 19 2012-07-01 2012-07-01 false Fuel flow measurement specifications....314-79 Fuel flow measurement specifications. (a) The fuel flow rate measurement instrument must have a... period of the clock. (2) For Diesel engines only, if the mass of fuel consumed is measured by...

  6. 40 CFR 91.417 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Fuel flow measurement specifications... Procedures § 91.417 Fuel flow measurement specifications. (a) Fuel flow measurement is required only for raw testing but is allowed for dilute testing. (b) The fuel flow rate measurement instrument must have...

  7. 40 CFR 89.414 - Air flow measurement specifications.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 20 2014-07-01 2013-07-01 true Air flow measurement specifications. 89... Test Procedures § 89.414 Air flow measurement specifications. (a) The air flow measurement method used... during the test. Overall measurement accuracy must be ± 2 percent of the maximum engine value for...

  8. 40 CFR 91.417 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 21 2012-07-01 2012-07-01 false Fuel flow measurement specifications... Procedures § 91.417 Fuel flow measurement specifications. (a) Fuel flow measurement is required only for raw testing but is allowed for dilute testing. (b) The fuel flow rate measurement instrument must have...

  9. 40 CFR 89.414 - Air flow measurement specifications.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 21 2013-07-01 2013-07-01 false Air flow measurement specifications... Emission Test Procedures § 89.414 Air flow measurement specifications. (a) The air flow measurement method... during the test. Overall measurement accuracy must be ± 2 percent of the maximum engine value for...

  10. 40 CFR 89.414 - Air flow measurement specifications.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 21 2012-07-01 2012-07-01 false Air flow measurement specifications... Emission Test Procedures § 89.414 Air flow measurement specifications. (a) The air flow measurement method... during the test. Overall measurement accuracy must be ± 2 percent of the maximum engine value for...

  11. The measurement of specific dynamic action in fishes.

    PubMed

    Chabot, D; Koenker, R; Farrell, A P

    2016-01-01

    Specific dynamic action (SDA) is the postprandial increase in oxygen uptake. Whereas it is easy to measure in fishes that remain calm and motionless during the entire digestion period, spontaneous locomotor activity is a frequent problem that leads to overestimation of SDA amplitude and magnitude (area under the curve, bound by the standard metabolic rate, SMR). Few studies have attempted to remove the effect of fish activity on SDA. A new method, non-parametric quantile regression, is described to estimate SDA even when pronounced circadian activity cycles are present. Data from juvenile Atlantic cod Gadus morhua are used to demonstrate its use and advantages compared with traditional techniques. Software (scripts in the R language) is provided to facilitate its use. PMID:26768974

  12. Photodynamic effect on specific antitumor immune activity

    NASA Astrophysics Data System (ADS)

    Vonarx-Coinsmann, Veronique; Foultier, Marie-Therese; Morlet, Laurent; de Brito, Leonor X.; Patrice, Thierry

    1995-03-01

    In this study the effect of PDT on the antitumoral specific immunologic response was evaluated. We compared the specific cytolytic activity (CLA) by a chromium release assay of primed mouse spleen T lymphocytes sensitized against syngeneic mastocytoma P511 cells. P511 cells, or lymphocytes, or both cells were treated or not with photofrin and/or light (514 nm). Photofrin II alone (1 (mu) g/ml, 2 hours) reduced CLA 59% when P511 were treated. Photofrin II (1 (mu) g/ml) followed by light (25 Joules/sq cm) also reduced CLA 35%. Photofrin II alone (0.5 (mu) g/ml, 2 hours) reduced CLA 8% when only lymphocytes were treated. And Photofrin II (0.5 (mu) g/ml) followed by light (25 Joules/sq cm) also reduced CLA 45%. When both cells were treated with Photofrin II alone or followed by light (25 Joules/sq cm) the CLA was also reduced respectively 19, 41%.

  13. Quantitative measurement of cytomegalovirus-specific IgG and IgM antibodies in relation to cytomegalovirus antigenaemia and disease activity in kidney recipients with an active cytomegalovirus infection.

    PubMed Central

    Van der Giessen, M; van den Berg, A P; van der Bij, W; Postma, S; van Son, W J; The, T H

    1990-01-01

    In a longitudinal investigation 103 kidney recipients were studied with respect to the development of cytomegalovirus (CMV) specific antibodies of the IgG and IgM class, in relation to the detection of CMV antigenaemia (immediate early antigen, IEA), in weekly obtained blood samples during the first 3 months after transplantation. In 15 out of 49 (31%) seronegative patients a primary infection occurred, which was characterized by a quick rise in IgM antibody followed by a slower production of IgG antibody, high maximum numbers of IEA+ cells, and a CMV syndrome in 11 patients. In 35 out of 54 (65%) seropositive patients a secondary infection occurred. After a post-operative fall in the IgG antibody, which was also found in patients without an active infection and which was accompanied by a similar drop in serum albumin and IgG, a second dip in IgG antibody was found 6 days before the first IEA+ leucocyte appeared in the blood. This was followed by a significant increase, indicative of an active immune response in consequence of the infection, 18 days later. In 31 of these 35 patients an IgM response was found. This could be ascribed to the presence of rheumatoid factor activity in 20 of them. Eight patients who showed a transient rise in IgG antibody between the two dips could be distinguished from the remaining ones by a lower maximum number of IEA+ cells and less severe disease symptoms. The described results suggest that (i) an adequate humoral immune system may prevent symptomatic CMV disease in secondary infections; and (ii) CMV-specific antibodies may be removed from the circulation by antigens present in infected tissues before CMV antigenaemia becomes detectable. PMID:2157567

  14. ACTIVITIES: Centimeter and Millimeter Measurements

    ERIC Educational Resources Information Center

    Bolster, L. Carey

    1974-01-01

    An activity is suggested which will give junior high school students practice in estimating and measuring in centimeters and millimeters. Three worksheets are given, one of which is a model for making a metric caliper. (LS)

  15. 40 CFR 90.417 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... specifications of 40 CFR part 1065, subpart C, instead of those in this paragraph (b). ... 40 Protection of Environment 21 2013-07-01 2013-07-01 false Fuel flow measurement specifications... Gaseous Exhaust Test Procedures § 90.417 Fuel flow measurement specifications. (a) Fuel flow...

  16. 40 CFR 90.417 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... specifications of 40 CFR part 1065, subpart C, instead of those in this paragraph (b). ... 40 Protection of Environment 20 2011-07-01 2011-07-01 false Fuel flow measurement specifications... Gaseous Exhaust Test Procedures § 90.417 Fuel flow measurement specifications. (a) Fuel flow...

  17. 40 CFR 90.417 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... specifications of 40 CFR part 1065, subpart C, instead of those in this paragraph (b). ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Fuel flow measurement specifications... Gaseous Exhaust Test Procedures § 90.417 Fuel flow measurement specifications. (a) Fuel flow...

  18. 40 CFR 90.417 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... specifications of 40 CFR part 1065, subpart C, instead of those in this paragraph (b). ... 40 Protection of Environment 21 2012-07-01 2012-07-01 false Fuel flow measurement specifications... Gaseous Exhaust Test Procedures § 90.417 Fuel flow measurement specifications. (a) Fuel flow...

  19. 40 CFR 89.414 - Air flow measurement specifications.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 20 2011-07-01 2011-07-01 false Air flow measurement specifications. 89.414 Section 89.414 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR... Emission Test Procedures § 89.414 Air flow measurement specifications. (a) The air flow measurement...

  20. 40 CFR 89.415 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 21 2013-07-01 2013-07-01 false Fuel flow measurement specifications... Emission Test Procedures § 89.415 Fuel flow measurement specifications. The fuel flow rate measurement instrument must have a minimum accuracy of 2 percent of the engine maximum fuel flow rate. The...

  1. 40 CFR 89.415 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 20 2011-07-01 2011-07-01 false Fuel flow measurement specifications... Emission Test Procedures § 89.415 Fuel flow measurement specifications. The fuel flow rate measurement instrument must have a minimum accuracy of 2 percent of the engine maximum fuel flow rate. The...

  2. 40 CFR 89.414 - Air flow measurement specifications.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Air flow measurement specifications. 89.414 Section 89.414 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR... Emission Test Procedures § 89.414 Air flow measurement specifications. (a) The air flow measurement...

  3. Production of high specific activity silicon-32

    DOEpatents

    Phillips, Dennis R.; Brzezinski, Mark A.

    1994-01-01

    A process for preparation of silicon-32 is provide and includes contacting an irradiated potassium chloride target, including spallation products from a prior irradiation, with sufficient water, hydrochloric acid or potassium hydroxide to form a solution, filtering the solution, adjusting pH of the solution to from about 5.5 to about 7.5, admixing sufficient molybdate-reagent to the solution to adjust the pH of the solution to about 1.5 and to form a silicon-molybdate complex, contacting the solution including the silicon-molybdate complex with a dextran-based material, washing the dextran-based material to remove residual contaminants such as sodium-22, separating the silicon-molybdate complex from the dextran-based material as another solution, adding sufficient hydrochloric acid and hydrogen peroxide to the solution to prevent reformation of the silicon-molybdate complex and to yield an oxidization state of the molybdate adapted for subsequent separation by an anion exchange material, contacting the solution with an anion exchange material whereby the molybdate is retained by the anion exchange material and the silicon remains in solution, and optionally adding sufficient alkali metal hydroxide to adjust the pH of the solution to about 12 to 13. Additionally, a high specific activity silicon-32 product having a high purity is provided.

  4. Production of high specific activity silicon-32

    SciTech Connect

    Phillips, D.R.; Brzezinski, M.A.

    1994-09-13

    A process for the preparation of silicon-32 is provided and includes contacting an irradiated potassium chloride target, including spallation products from a prior irradiation, with sufficient water, hydrochloric acid or potassium hydroxide to form a solution, filtering the solution, adjusting pH of the solution to from about 5.5 to about 7.5, admixing sufficient molybdate-reagent to the solution to adjust the pH of the solution to about 1.5 and to form a silicon-molybdate complex, contacting the solution including the silicon-molybdate complex with a dextran-based material, washing the dextran-based material to remove residual contaminants such as sodium-22, separating the silicon-molybdate complex from the dextran-based material as another solution, adding sufficient hydrochloric acid and hydrogen peroxide to the solution to prevent reformation of the silicon-molybdate complex and to yield an oxidization state of the molybdate adapted for subsequent separation by an anion exchange material, contacting the solution with an anion exchange material whereby the molybdate is retained by the anion exchange material and the silicon remains in solution, and optionally adding sufficient alkali metal hydroxide to adjust the pH of the solution to about 12 to 13. Additionally, a high specific activity silicon-32 product having a high purity is provided.

  5. 40 CFR 90.416 - Intake air flow measurement specifications.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Intake air flow measurement... Gaseous Exhaust Test Procedures § 90.416 Intake air flow measurement specifications. (a) If used, the engine intake air flow measurement method used must have a range large enough to accurately measure...

  6. 40 CFR 91.416 - Intake air flow measurement specifications.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Intake air flow measurement... Procedures § 91.416 Intake air flow measurement specifications. (a) If used, the engine intake air flow measurement method used must have a range large enough to accurately measure the air flow over the...

  7. Measuring Offence-Specific Forgiveness in Marriage: The Marital Offence-Specific Forgiveness Scale (MOFS)

    ERIC Educational Resources Information Center

    Paleari, F. Giorgia; Regalia, Camillo; Fincham, Frank D.

    2009-01-01

    Three studies involving 328 married couples were conducted to validate the Marital Offence-Specific Forgiveness Scale, a new measure assessing offence-specific forgiveness for marital transgressions. The studies examined the dimensionality; internal consistency; and discriminant, concurrent, and predictive validity of the new measure. The final…

  8. 10 CFR 810.8 - Activities requiring specific authorization.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 4 2014-01-01 2014-01-01 false Activities requiring specific authorization. 810.8 Section 810.8 Energy DEPARTMENT OF ENERGY ASSISTANCE TO FOREIGN ATOMIC ENERGY ACTIVITIES § 810.8 Activities requiring specific authorization. Unless generally authorized by § 810.7, a person requires specific authorization by the Secretary of Energy...

  9. Stage-specific fucosyltransferase activity during mouse spermatogenesis

    SciTech Connect

    Cardullo, R.A.; Armant, D.R.; Millette, C.F.

    1986-05-01

    This laboratory is involved in the biochemical characterization of developing spermatogenic cells. The authors have measured the in vitro activity of fucosyltransferase (FT) in germ cells. FT activity was assayed with a procedure modified from Letts et al. using GDP-(/sup 14/C)-fucose and asialofetuin as substrates. After incubation for 15 minutes at 33/sup 0/C, the reaction was stopped by adding cold 500 mM EDTA. Radiolabeled asialofetuin was isolated using Bio-Gel P-10 chromatography. The FT activity of germ cells purified from seminiferous tubules was 18.5 +/- 1.7 pmol/mg protein-min. To see if this activity varied at different stages of development, germ cells were further separated in a STAPUT chamber using a 2-4% BSA gradient. Pachytene spermatocytes or round spermatids were purified to at least 87%. The FT activity in isolated pachytene spermatocytes was 24.4 +/- 1.2 pmol/mg protein-min while the activity in isolated round spermatids was 49.0 +/- 7.2 pmol/mg protein-min. These results suggest that the highest FT activity is in developing spermatogenic cells with round spermatids having nearly twice the FT activity as pachytene spermatocytes. This increase in FT activity may be biologically significant since it occurs at a time when the Golgi apparatus is undergoing differentiation and when stage-specific fucosylated proteins appear.

  10. Site–Specific Monoubiquitination Activates Ras by Impeding GTPase Activating Protein Function

    PubMed Central

    Baker, Rachael; Lewis, Steven M.; Sasaki, Atsuo T.; Wilkerson, Emily M.; Locasale, Jason W.; Cantley, Lewis C.; Kuhlman, Brian; Dohlman, Henrik G.; Campbell, Sharon L.

    2012-01-01

    SUMMARY Cell growth and differentiation are controlled by growth factor receptors coupled to the GTPase Ras. Oncogenic mutations disrupt GTPase activity leading to persistent Ras signaling and cancer progression. Recent evidence indicates that monoubiquitination of Ras leads to Ras activation. Mutation of the primary site of monoubiquitination impairs the ability of activated K–Ras to promote tumor growth. To determine the mechanism of human Ras activation we chemically ubiquitinated the protein and analyzed its function by NMR, computational modeling, and biochemical activity measurements. We established that monoubiquitination has little effect on Ras GTP binding, GTP hydrolysis, or exchange factor activation, but severely abrogates the response to GTPase activating proteins in a site–specific manner. These findings reveal a new mechanism by which Ras can trigger persistent signaling in the absence of receptor activation or an oncogenic mutation. PMID:23178454

  11. 10 CFR 810.8 - Activities requiring specific authorization.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 4 2010-01-01 2010-01-01 false Activities requiring specific authorization. 810.8 Section 810.8 Energy DEPARTMENT OF ENERGY ASSISTANCE TO FOREIGN ATOMIC ENERGY ACTIVITIES § 810.8 Activities requiring specific authorization. Unless generally authorized by § 810.7, a person requires...

  12. 10 CFR 810.8 - Activities requiring specific authorization.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 10 Energy 4 2011-01-01 2011-01-01 false Activities requiring specific authorization. 810.8 Section 810.8 Energy DEPARTMENT OF ENERGY ASSISTANCE TO FOREIGN ATOMIC ENERGY ACTIVITIES § 810.8 Activities requiring specific authorization. Unless generally authorized by § 810.7, a person requires...

  13. 40 CFR 90.417 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... specifications of 40 CFR part 1065, subpart C, instead of those in this paragraph (b). ... measurement instrument must have a minimum accuracy of one percent of full-scale flow rate for each... ±five percent of full-scale flow rate for the measurement range used. The controlling parameters are...

  14. 40 CFR 1065.205 - Performance specifications for measurement instruments.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 33 2011-07-01 2011-07-01 false Performance specifications for measurement instruments. 1065.205 Section 1065.205 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR POLLUTION CONTROLS ENGINE-TESTING PROCEDURES Measurement Instruments § 1065.205...

  15. 40 CFR 1065.205 - Performance specifications for measurement instruments.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 32 2010-07-01 2010-07-01 false Performance specifications for measurement instruments. 1065.205 Section 1065.205 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR POLLUTION CONTROLS ENGINE-TESTING PROCEDURES Measurement Instruments § 1065.205...

  16. 40 CFR 1065.205 - Performance specifications for measurement instruments.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 34 2012-07-01 2012-07-01 false Performance specifications for measurement instruments. 1065.205 Section 1065.205 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR POLLUTION CONTROLS ENGINE-TESTING PROCEDURES Measurement Instruments § 1065.205...

  17. 40 CFR 1065.205 - Performance specifications for measurement instruments.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 34 2013-07-01 2013-07-01 false Performance specifications for measurement instruments. 1065.205 Section 1065.205 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR POLLUTION CONTROLS ENGINE-TESTING PROCEDURES Measurement Instruments § 1065.205...

  18. Slice-Based Formal Specification Measures -- Mapping Coupling and Cohesion Measures to Formal Z

    NASA Technical Reports Server (NTRS)

    Bollin, Andreas

    2010-01-01

    This paper demonstrates that existing slice-based measures can reasonably be mapped to the field of state-based specification languages. By making use of Z specifications this contribution renews the idea of slice-profiles and derives coupling and cohesion measures for them. The measures are then assessed by taking a critical look at their sensitiveness in respect to modifications on the specification source. The presented study shows that slice-based coupling and cohesion measures have the potential to be used as quality indicators for specifications as they reflect the changes in the structure of a specification as accustomed from their program-related pendants.

  19. Simultaneous specific heat and thermal conductivity measurement of individual nanostructures

    NASA Astrophysics Data System (ADS)

    Zheng, Jianlin; Wingert, Matthew C.; Moon, Jaeyun; Chen, Renkun

    2016-08-01

    Fundamental phonon transport properties in semiconductor nanostructures are important for their applications in energy conversion and storage, such as thermoelectrics and photovoltaics. Thermal conductivity measurements of semiconductor nanostructures have been extensively pursued and have enhanced our understanding of phonon transport physics. Specific heat of individual nanostructures, despite being an important thermophysical parameter that reflects the thermodynamics of solids, has remained difficult to characterize. Prior measurements were limited to ensembles of nanostructures in which coupling and sample inhomogeneity could play a role. Herein we report the first simultaneous specific heat and thermal conductivity measurements of individual rod-like nanostructures such as nanowires and nanofibers. This technique is demonstrated by measuring the specific heat and thermal conductivity of single ∼600–700 nm diameter Nylon-11 nanofibers (NFs). The results show that the thermal conductivity of the NF is increased by 50% over the bulk value, while the specific heat of the NFs exhibits bulk-like behavior. We find that the thermal diffusivity obtained from the measurement, which is related to the phonon mean free path (MFP), decreases with temperature, indicating that the intrinsic phonon Umklapp scattering plays a role in the NFs. This platform can also be applied to one- and two- dimensional semiconductor nanostructures to probe size effects on the phonon spectra and other transport physics.

  20. Status of LDEF activation measurements and archive

    NASA Technical Reports Server (NTRS)

    Harmon, B. Alan; Parnell, Thomas A.; Laird, Christopher E.

    1995-01-01

    We review the status of induced radioactivity measurements for the LDEF spacecraft which includes studies of the nuclide, target, directional and depth dependences of the activation. Analysis of the data has focused on extraction of the specific activities for many materials to develop a global picture of the low Earth orbital environment to which the LDEF was subjected. Preliminary comparisons of data in a previous review showed that it was possible to make meaningful intercomparisons between results obtained at different facilities. Generally these comparisons were good and gave results to within 10-20 percent, although some analysis remains. These results clearly provide constraints for recent calculations being performed of the radiation environment of the LDEF. We are not anticipating a period of production of final activation results. An archive is being prepared jointly between NASA/Marshall and Eastern Kentucky University which will include gamma ray spectra and other intermediate results.

  1. 50 CFR 660.60 - Specifications and management measures.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 13 2012-10-01 2012-10-01 false Specifications and management measures. 660.60 Section 660.60 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE (CONTINUED) FISHERIES OFF WEST COAST STATES West Coast Groundfish Fisheries §...

  2. 50 CFR 660.60 - Specifications and management measures.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... regulations at 50 CFR part § 600.745 for limited testing, public display, data collection, exploratory, health.... 660.60 Section 660.60 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND... Groundfish Fisheries § 660.60 Specifications and management measures. (a) General. NMFS will establish...

  3. 50 CFR 660.60 - Specifications and management measures.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... regulations at 50 CFR part § 600.745 for limited testing, public display, data collection, exploratory, health.... 660.60 Section 660.60 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND... Groundfish Fisheries § 660.60 Specifications and management measures. (a) General. NMFS will establish...

  4. 50 CFR 660.60 - Specifications and management measures.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... prohibited for retention. (2) The Regional Administrator may also issue EFPs under regulations at 50 CFR part.... 660.60 Section 660.60 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND... Groundfish Fisheries § 660.60 Specifications and management measures. (a) General. NMFS will establish...

  5. 50 CFR 660.60 - Specifications and management measures.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... prohibited for retention. (2) The Regional Administrator may also issue EFPs under regulations at 50 CFR part.... 660.60 Section 660.60 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND... Groundfish Fisheries § 660.60 Specifications and management measures. (a) General. NMFS will establish...

  6. 50 CFR 660.370 - Specifications and management measures.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 50 Wildlife and Fisheries 9 2010-10-01 2010-10-01 false Specifications and management measures. 660.370 Section 660.370 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE (CONTINUED) FISHERIES OFF WEST COAST STATES West Coast Groundfish Fisheries §...

  7. Measurement of glutathione in activated sludges.

    PubMed

    Dziurla, M A; Leroy, P; Strünkmann, G W; Salhi, M; Lee, D U; Camacho, P; Heinz, V; Müller, J A; Paul, E; Ginestet, Ph; Audic, J M; Block, J C

    2004-01-01

    Thermal, electric, mechanical or oxidative stress seem a promising way to reduce the production of excess activated sludge during biological wastewater treatment. However, the adaptation and the resistance of the sludge microbial ecosystem to stress conditions is a major question as it may definitively limit the effect of some treatments. Defence mechanisms developed by aerobic organisms, in particular, in response to oxidative stress involve various antioxidant activities and compounds such as glutathione. An HPLC method was developed for measuring reduced and total glutathione (GSH and GSHt) in perchloric acid sludge extracts. The method was sensitive, highly specific and validated for linearity, precision and recovery. Considering the extraction yield and the oxidation of GSH during extract storage, the measured GSH concentration was estimated to represent 60% of the GSH content from activated sludges. GSHt ranged from 0.32 to 3.34micromolg(-1) volatile solids and the GSH/GSHt ratio ranged from 32% to 91%. Measurements performed on sludges stressed in precise conditions selected to reach a reduction of sludge production showed a decrease of GSH and GSHt concentrations with thermal, mechanical, electric and ozone stress. PMID:14630122

  8. Suppression of Antigen-Specific Lymphocyte Activation in Simulated Microgravity

    NASA Technical Reports Server (NTRS)

    Cooper, David; Pride, Michael W.; Brown, Eric L.; Risin, Diana; Pellis, Neal R.

    1999-01-01

    Various parameters of immune suppression are observed in astronauts during and after spaceflight, and in isolated immune cells in true and simulated microgravity. Specifically, polyclonal activation of T cells is severely suppressed in true and simulated microgravity. These recent findings with various polyclonal activators suggests a suppression of oligoclonal lymphocyte activation in microgravity. We utilized rotating wall vessel (RWV) bioreactors that simulate aspects of microgravity for cell cultures to analyze three models of antigen-specific activation. A mixed-lymphocyte reaction (MLR), as a model for a primary immune response; a tetanus toxoid (TT) response and a B. burgdorferi (Bb) response, as models of a secondary immune response, were all suppressed in the RWV bioreactor. Our findings confirm that the suppression of activation observed with polyclonal models also encompasses oligoclonal antigen-specific activation.

  9. An evaluation of the effect of age and the peri-parturient period on bone metabolism in dairy cows as measured by serum bone-specific alkaline phosphatase activity and urinary deoxypyridinoline concentration.

    PubMed

    Sato, Reiichiro; Onda, Ken; Kato, Hajime; Ochiai, Hideharu; Kawai, Kazuhiro; Iriki, Tsunenori; Kaneko, Kazuyuki; Yamazaki, Yukio; Wada, Yasunori

    2013-08-01

    Various biochemical markers help to evaluate the state of bone turnover in humans and could be used during the peri-parturient period in dairy cows when calcium (Ca) metabolism changes dramatically. To investigate this, the peri-partum characteristics of serum bone-specific alkaline phosphatase (BAP) and urinary deoxypyridinoline (DPD) were investigated. Both serum BAP activity and urinary DPD concentrations were increased and demonstrated wide variability in younger animals, and these findings were consistent with other bone turnover markers. Around the time of parturition, serum Ca concentration and serum BAP activity in multiparous cows were significantly lower than in primiparous cows, but urinary DPD concentration was unchanged. The use of BAP as a bone formation marker appears to be valuable for evaluating bone remodelling status in cows, but the specificity of the test needs to be confirmed. The DPD/BAP ratio around parturition demonstrated a clear difference in bone turnover status between the two parity groups with multiparous cows demonstrating increased signs of bone resorption compared with primiparous cows, corresponding to the Ca requirement for milk production. In future studies, the applicability of the ratio of bone resorption marker to bone formation marker should be evaluated for bone turnover assessment. PMID:23422881

  10. Activities-Specific Balance Confidence in People with Multiple Sclerosis

    PubMed Central

    Nilsagård, Ylva; Carling, Anna; Forsberg, Anette

    2012-01-01

    Objective. To evaluate the validity of the Activities-specific Balance Confidence scale (ABC) in people with multiple sclerosis (PwMS). Design. A multicentre, cross-sectional study. Setting. Six rural and urban Swedish sites, including specialized units at hospitals and primary care centers. Participants. A sample of 84 PwMS with subjective gait and balance impairment but still able to walk 100 m (comparable with EDSS 1–6). Outcome Measures. Timed Up and Go, Timed Up and Gocog, 25-foot Timed Walk Test, Four Square Step Test, Dynamic Gait Index, Chair Stand Test, 12-item MS Walking Scale, self-reported falls, and use of assistive walking device were used for validation. Results. The concurrent convergent validity was moderate to good (0.50 to −0.75) with the highest correlation found for the 12-item MS Walking Scale. The ABC discriminated between multiple fallers and nonfallers but not between men and women. Ecological validity is suggested since ABC discriminated between users of assistive walking device and nonusers. The internal consistency was high at α = 0.95, and interitem correlations were between 0.30 and 0.83. Conclusion. This study supports the validity of the ABC for persons with mild-to-moderate MS. The participants lacked balance confidence in many everyday activities, likely restricting their participation in society. PMID:22919491

  11. A specific, transmembrane interface regulates fibroblast activation protein (FAP) homodimerization, trafficking and exopeptidase activity.

    PubMed

    Wonganu, Benjamaporn; Berger, Bryan W

    2016-08-01

    Fibroblast activation protein (FAP) is a cell-surface serine protease which promotes invasiveness of certain epithelial cancers and is therefore a potential target for cancer drug development and delivery. Unlike dipeptidyl peptidase IV (DPPIV), FAP exhibits prolyl endopeptidase activity and is active as a homodimer with specificity for type I collagen. The mechanism that regulates FAP homodimerization and its relation to prolyl endopeptidase activity is not completely understood. Here, we investigate key residues in the FAP TM domain that may be significant for FAP homodimerization. Mutations to predicted TM interfacial residues (G10L, S14L, and A18L) comprising a small-X3-small motif reduced FAP TM-CYTO dimerization relative to wild type as measured using the AraTM assay, whereas predicted off-interface residues showed no significant change from wild type. The results implied that the predicted small-X3-small dimer interface affect stabilization of FAP TM-CYTO homodimerization. Compared with FAPwild-type, the interfacial TM residue G10L significantly decreased FAP endopeptidase activity more than 25%, and also reduced cell-surface versus intracellular expression relative to other interfacial residues S14L and A18L. Thus, our results suggest FAP dimerization is important for both trafficking and protease activity, and is dependent on a specific TM interface. PMID:27155568

  12. Phase Measurement of Cognitive Impairment Specific to Radiotherapy

    SciTech Connect

    Armstrong, Carol L.; Shera, David M.; Lustig, Robert A.; Phillips, Peter C.

    2012-07-01

    Purpose: Memory impairment is an early-delayed effect of radiotherapy (RT). The prospective longitudinal measurement of the cognitive phase effects from RT was conducted on treated and untreated brain tumor patients. The study design investigated semantic vs. perceptual and visual vs. verbal memory to determine the most disease-specific measure of RT-related changes and understanding of the neurotoxicity from RT to the brain. Methods and Materials: Tests of memory that had previously shown RT-related phasic changes were compared with experimental tests of memory to test hypotheses about cognition targeted to the neural toxicity of RT. The results from 41 irradiated and 29 nonirradiated patients with low-grade, supratentorial tumors were analyzed. The methods controlled for comorbid white matter risk, recurrence, interval after treatment, and age (18-69 years). The effects were examined before RT and at three points after RT to 1 year using a mixed effects model that included interval, group, surgical status, medication use, practice, and individual random effects. Four new tests of memory and other candidate cognitive tests were investigated, and a post hoc analysis of a comprehensive battery of tests was performed to identify the cognitive processes most specific to RT. Results: The RT effects on memory were identified in the treated group only; among the new tests of memory and the complete neurocognitive battery, the RT effects were significant only for delayed recall (p < 0.009) and interval to recognize (p < 0.002). Tumor location was not related to the treatment effect. Memory decline was specific to retrieval of semantic memories; a double dissociation of semantic from perceptual visual memory was demonstrated in the RT group. Conclusions: These results implicate memory dependent on the semantic cortex and the hippocampal memory system. A cognitive measurement that is brief but specific to neural mechanisms is effective and feasible for studies of RT damage.

  13. UML activity diagrams in requirements specification of logic controllers

    NASA Astrophysics Data System (ADS)

    Grobelna, Iwona; Grobelny, Michał

    2015-12-01

    Logic controller specification can be prepared using various techniques. One of them is the wide understandable and user-friendly UML language and its activity diagrams. Using formal methods during the design phase increases the assurance that implemented system meets the project requirements. In the approach we use the model checking technique to formally verify a specification against user-defined behavioral requirements. The properties are usually defined as temporal logic formulas. In the paper we propose to use UML activity diagrams in requirements definition and then to formalize them as temporal logic formulas. As a result, UML activity diagrams can be used both for logic controller specification and for requirements definition, what simplifies the specification and verification process.

  14. Mechanism of dual specificity kinase activity of DYRK1A.

    PubMed

    Walte, Agnes; Rüben, Katharina; Birner-Gruenberger, Ruth; Preisinger, Christian; Bamberg-Lemper, Simone; Hilz, Nikolaus; Bracher, Franz; Becker, Walter

    2013-09-01

    The function of many protein kinases is controlled by the phosphorylation of a critical tyrosine residue in the activation loop. Dual specificity tyrosine-phosphorylation-regulated kinases (DYRKs) autophosphorylate on this tyrosine residue but phosphorylate substrates on aliphatic amino acids. This study addresses the mechanism of dual specificity kinase activity in DYRK1A and related kinases. Tyrosine autophosphorylation of DYRK1A occurred rapidly during in vitro translation and did not depend on the non-catalytic domains or other proteins. Expression in bacteria as well as in mammalian cells revealed that tyrosine kinase activity of DYRK1A is not restricted to the co-translational autophosphorylation in the activation loop. Moreover, mature DYRK1A was still capable of tyrosine autophosphorylation. Point mutants of DYRK1A and DYRK2 lacking the activation loop tyrosine showed enhanced tyrosine kinase activity. A series of structurally diverse DYRK1A inhibitors was used to pharmacologically distinguish different conformational states of the catalytic domain that are hypothesized to account for the dual specificity kinase activity. All tested compounds inhibited substrate phosphorylation with higher potency than autophosphorylation but none of the tested inhibitors differentially inhibited threonine and tyrosine kinase activity. Finally, the related cyclin-dependent kinase-like kinases (CLKs), which lack the activation loop tyrosine, autophosphorylated on tyrosine both in vitro and in living cells. We propose a model of DYRK autoactivation in which tyrosine autophosphorylation in the activation loop stabilizes a conformation of the catalytic domain with enhanced serine/threonine kinase activity without disabling tyrosine phosphorylation. The mechanism of dual specificity kinase activity probably applies to related serine/threonine kinases that depend on tyrosine autophosphorylation for maturation. PMID:23809146

  15. Specific activation of the thyrotropin receptor by trypsin.

    PubMed

    Van Sande, J; Massart, C; Costagliola, S; Allgeier, A; Cetani, F; Vassart, G; Dumont, J E

    1996-05-31

    The identification of 16 different activating mutations in the TSH receptor, found in patients suffering from toxic autonomous adenomas or congenital hyperthyroidism, leads to the concept that this receptor is in a constrained conformation in its wild-type form. We used mild trypsin treatment of CHO-K1 cells or COS-7 cells, stably or transiently transfected with the human TSH receptor, respectively, and measured its consequences on the TSH receptor coupled cascades, i.e. cyclic AMP and inositol-phosphates accumulation. A 2-min, 0.01% trypsin treatment increased stably cyclic AMP but not inositol-phosphates formation. This was not observed after chymotrypsin, thrombin and endoproteinase glu C treatment. The TSH action on cyclic AMP was decreased by only 25%. The effect was also observed in cells expressing the dog TSH receptor. It was not observed in MSH receptor, LH receptor expressing or mock transfected cells (vector alone). It is therefore specific for the TSH receptor, for its action on the Gs/adenylate cyclase cascade, and for the proteolytic cleavage caused by trypsin. Using monoclonal (A. Johnstone and P. Shepherd, personal communication) and polyclonal antibodies directed against the extracellular domain of the TSH receptor, it was shown that treatment by trypsin removes or destroys a VFFEEQ epitope (residues 354-359) from the receptor. The effect mimics the action of TSH as it activates Gs alpha and enhances the action of forskolin. It is not reversible in 1 h. The results support the concept that activation of the receptor (by hormone, autoantibodies, mutations or mild proteolysis) might involve the relief of a built-in negative constrain. They suggest that the C-terminal portion of the large extracellular domain plays a role in the maintenance of this constrain. PMID:8807635

  16. Measuring the specific contact resistance of contacts to semiconductor nanowires

    NASA Astrophysics Data System (ADS)

    Mohney, S. E.; Wang, Y.; Cabassi, M. A.; Lew, K. K.; Dey, S.; Redwing, J. M.; Mayer, T. S.

    2005-02-01

    Ohmic contacts to semiconductor nanowires are essential components of many new nanoscale electronic devices. Equations for extracting specific contact resistance (or contact resistivity) from several different test structures have been developed by modeling the metal/semiconductor contact as a transmission line, leading to the development of equations analogous to those used for planar contacts. The advantages and disadvantages of various test structures are discussed. To fabricate test structures using a convenient four-point approach, silicon nanowires have been aligned using field-assisted assembly and contacts fabricated. Finally, specific contact resistances near 5 × 10 -4 Ω cm 2 have been measured for Ti/Au contacts to p-type Si nanowires with diameters of 78 and 104 nm.

  17. Measurement of Spray Drift with a Specifically Designed Lidar System.

    PubMed

    Gregorio, Eduard; Torrent, Xavier; Planas de Martí, Santiago; Solanelles, Francesc; Sanz, Ricardo; Rocadenbosch, Francesc; Masip, Joan; Ribes-Dasi, Manel; Rosell-Polo, Joan R

    2016-01-01

    Field measurements of spray drift are usually carried out by passive collectors and tracers. However, these methods are labour- and time-intensive and only provide point- and time-integrated measurements. Unlike these methods, the light detection and ranging (lidar) technique allows real-time measurements, obtaining information with temporal and spatial resolution. Recently, the authors have developed the first eye-safe lidar system specifically designed for spray drift monitoring. This prototype is based on a 1534 nm erbium-doped glass laser and an 80 mm diameter telescope, has scanning capability, and is easily transportable. This paper presents the results of the first experimental campaign carried out with this instrument. High coefficients of determination (R² > 0.85) were observed by comparing lidar measurements of the spray drift with those obtained by horizontal collectors. Furthermore, the lidar system allowed an assessment of the drift reduction potential (DRP) when comparing low-drift nozzles with standard ones, resulting in a DRP of 57% (preliminary result) for the tested nozzles. The lidar system was also used for monitoring the evolution of the spray flux over the canopy and to generate 2-D images of these plumes. The developed instrument is an advantageous alternative to passive collectors and opens the possibility of new methods for field measurement of spray drift. PMID:27070613

  18. Measurement of Spray Drift with a Specifically Designed Lidar System

    PubMed Central

    Gregorio, Eduard; Torrent, Xavier; Planas de Martí, Santiago; Solanelles, Francesc; Sanz, Ricardo; Rocadenbosch, Francesc; Masip, Joan; Ribes-Dasi, Manel; Rosell-Polo, Joan R.

    2016-01-01

    Field measurements of spray drift are usually carried out by passive collectors and tracers. However, these methods are labour- and time-intensive and only provide point- and time-integrated measurements. Unlike these methods, the light detection and ranging (lidar) technique allows real-time measurements, obtaining information with temporal and spatial resolution. Recently, the authors have developed the first eye-safe lidar system specifically designed for spray drift monitoring. This prototype is based on a 1534 nm erbium-doped glass laser and an 80 mm diameter telescope, has scanning capability, and is easily transportable. This paper presents the results of the first experimental campaign carried out with this instrument. High coefficients of determination (R2 > 0.85) were observed by comparing lidar measurements of the spray drift with those obtained by horizontal collectors. Furthermore, the lidar system allowed an assessment of the drift reduction potential (DRP) when comparing low-drift nozzles with standard ones, resulting in a DRP of 57% (preliminary result) for the tested nozzles. The lidar system was also used for monitoring the evolution of the spray flux over the canopy and to generate 2-D images of these plumes. The developed instrument is an advantageous alternative to passive collectors and opens the possibility of new methods for field measurement of spray drift. PMID:27070613

  19. Measurement of Specific Heat Capacity Using Differential Scanning Calorimeter

    SciTech Connect

    J. E. Daw

    2008-11-01

    This document describes the process used at the Idaho National Laboratory’s (INL) High Temperature Test Laboratory (HTTL) for measuring specific heat capacity using a differential scanning calorimeter (DSC). The document is divided into four sections: Approach, in which the technique is described; Setup, in which the physical system is described; Procedure, in which the testing steps are listed and detailed; and Example Test, in which a typical test is outlined following the steps listed in the Procedure section. Example data, results, photos, and curves are provided throughout the document to assist other users of this system.

  20. Activation energy measurements of cheese

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Temperature sweeps of cheeses using small amplitude oscillatory shear tests produced values for activation energy of flow (Ea) between 30 and 44 deg C. Soft goat cheese and Queso Fresco, which are high-moisture cheeses and do not flow when heated, exhibited Ea values between 30 and 60 kJ/mol. The ...

  1. Energy Landscape Topography Reveals the Underlying Link Between Binding Specificity and Activity of Enzymes.

    PubMed

    Chu, Wen-Ting; Wang, Jin

    2016-01-01

    Enzyme activity (often quantified by kcat/Km) is the main function of enzyme when it is active against the specific substrate. Higher or lower activities are highly desired for the design of novel enzyme and drug resistance. However, it is difficult to measure the activities of all possible variants and find the "hot-spot" within the limit of experimental time. In this study, we explore the underlying energy landscape of enzyme-substrate interactions and introduce the intrinsic specificity ratio (ISR), which reflects the landscape topography. By studying two concrete systems, we uncover the statistical correlation between the intrinsic specificity and the enzyme activity kcat/Km. This physics-based concept and method show that the energy landscape topography is valuable for understanding the relationship between enzyme specificity and activity. In addition, it can reveal the underlying mechanism of enzyme-substrate actions and has potential applications on enzyme design. PMID:27298067

  2. Energy Landscape Topography Reveals the Underlying Link Between Binding Specificity and Activity of Enzymes

    PubMed Central

    Chu, Wen-Ting; Wang, Jin

    2016-01-01

    Enzyme activity (often quantified by kcat/Km) is the main function of enzyme when it is active against the specific substrate. Higher or lower activities are highly desired for the design of novel enzyme and drug resistance. However, it is difficult to measure the activities of all possible variants and find the “hot-spot” within the limit of experimental time. In this study, we explore the underlying energy landscape of enzyme-substrate interactions and introduce the intrinsic specificity ratio (ISR), which reflects the landscape topography. By studying two concrete systems, we uncover the statistical correlation between the intrinsic specificity and the enzyme activity kcat/Km. This physics-based concept and method show that the energy landscape topography is valuable for understanding the relationship between enzyme specificity and activity. In addition, it can reveal the underlying mechanism of enzyme-substrate actions and has potential applications on enzyme design. PMID:27298067

  3. Energy Landscape Topography Reveals the Underlying Link Between Binding Specificity and Activity of Enzymes

    NASA Astrophysics Data System (ADS)

    Chu, Wen-Ting; Wang, Jin

    2016-06-01

    Enzyme activity (often quantified by kcat/Km) is the main function of enzyme when it is active against the specific substrate. Higher or lower activities are highly desired for the design of novel enzyme and drug resistance. However, it is difficult to measure the activities of all possible variants and find the “hot-spot” within the limit of experimental time. In this study, we explore the underlying energy landscape of enzyme-substrate interactions and introduce the intrinsic specificity ratio (ISR), which reflects the landscape topography. By studying two concrete systems, we uncover the statistical correlation between the intrinsic specificity and the enzyme activity kcat/Km. This physics-based concept and method show that the energy landscape topography is valuable for understanding the relationship between enzyme specificity and activity. In addition, it can reveal the underlying mechanism of enzyme-substrate actions and has potential applications on enzyme design.

  4. Measurement of the Specific Heat Using a Gravity Cancellation Approach

    NASA Technical Reports Server (NTRS)

    Zhong, Fang

    2003-01-01

    The specific heat at constant volume C(sob V) of a simple fluid diverges near its liquid-vapor critical point. However, gravity-induced density stratification due to the divergence of isothermal susceptibility hinders the direct comparison of the experimental data with the predictions of renormalization group theory. In the past, a microgravity environment has been considered essential to eliminate the density stratification. We propose to perform specific heat measurements of He-3 on the ground using a method to cancel the density stratification. A He-3 fluid layer will be heated from below, using the thermal expansion of the fluid to cancel the hydrostatic compression. A 6% density stratification at a reduced temperature of 10(exp -5) can be cancelled to better than 0.1% with a steady 1.7 micro K temperature difference across a 0.05 cm thick fluid layer. A conventional AC calorimetry technique will be used to determine the heat capacity. The minimized bulk density stratification with a relaxation time 6500 sec at a reduced temperature of 10(exp -5) will stay unchanged during 1 Hz AC heating. The smear of the specific heat divergence due to the temperature difference across the cell is about 0.1% at a reduced temperature of 10(exp -6). The combination of using High Resolution Thermometry with a 0.5 n K temperature resolution in the AC technique and the cancellation of the density stratification will enable C(sub V) to be measured down to a reduced temperature of 10(exp -6) with less than a 1% systematic error.

  5. Suppression of antigen-specific lymphocyte activation in modeled microgravity

    NASA Technical Reports Server (NTRS)

    Cooper, D.; Pride, M. W.; Brown, E. L.; Risin, D.; Pellis, N. R.; McIntire, L. V. (Principal Investigator)

    2001-01-01

    Various parameters of immune suppression are observed in lymphocytes from astronauts during and after a space flight. It is difficult to ascribe this suppression to microgravity effects on immune cells in crew specimens, due to the complex physiological response to space flight and the resultant effect on in vitro immune performance. Use of isolated immune cells in true and modeled microgravity in immune performance tests, suggests a direct effect of microgravity on in vitro cellular function. Specifically, polyclonal activation of T-cells is severely suppressed in true and modeled microgravity. These recent findings suggest a potential suppression of oligoclonal antigen-specific lymphocyte activation in microgravity. We utilized rotating wall vessel (RWV) bioreactors as an analog of microgravity for cell cultures to analyze three models of antigen-specific activation. A mixed-lymphocyte reaction, as a model for a primary immune response, a tetanus toxoid response and a Borrelia burgdorferi response, as models of a secondary immune response, were all suppressed in the RWV bioreactor. Our findings confirm that the suppression of activation observed with polyclonal models also encompasses oligoclonal antigen-specific activation.

  6. High efficiency cell-specific targeting of cytokine activity

    NASA Astrophysics Data System (ADS)

    Garcin, Geneviève; Paul, Franciane; Staufenbiel, Markus; Bordat, Yann; van der Heyden, José; Wilmes, Stephan; Cartron, Guillaume; Apparailly, Florence; de Koker, Stefaan; Piehler, Jacob; Tavernier, Jan; Uzé, Gilles

    2014-01-01

    Systemic toxicity currently prevents exploiting the huge potential of many cytokines for medical applications. Here we present a novel strategy to engineer immunocytokines with very high targeting efficacies. The method lies in the use of mutants of toxic cytokines that markedly reduce their receptor-binding affinities, and that are thus rendered essentially inactive. Upon fusion to nanobodies specifically binding to marker proteins, activity of these cytokines is selectively restored for cell populations expressing this marker. This ‘activity-by-targeting’ concept was validated for type I interferons and leptin. In the case of interferon, activity can be directed to target cells in vitro and to selected cell populations in mice, with up to 1,000-fold increased specific activity. This targeting strategy holds promise to revitalize the clinical potential of many cytokines.

  7. Youth Physical Activity Resources Use and Activity Measured by Accelerometry

    PubMed Central

    Maslow, Andréa L.; Colabianchi, Natalie

    2014-01-01

    Objectives To examine whether utilization of physical activity resources (eg, parks) was associated with daily physical activity measured by accelerometry. Methods 111 adolescents completed a travel diary with concurrent accelerometry. The main exposure was self-reported utilization of a physical activity resource (none/1+ resources). The main outcomes were total minutes spent in daily 1) moderate-vigorous physical activity and 2) vigorous physical activity. Results Utilizing a physical activity resource was significantly associated with total minutes in moderate-vigorous physical activity. African-Americans and males had significantly greater moderate-vigorous physical activity. Conclusions Results from this study support the development and use of physical activity resources. PMID:21204684

  8. Development of a Trauma-Specific Quality of Life Measurement

    PubMed Central

    Wanner, John Paul; deRoon-Cassini, Terri; Kodadek, Lisa; Brasel, Karen

    2016-01-01

    Background Complex, disease-specific factors help to determine post-traumatic quality of life, but current practice utilizes outcome measures generated from the general population. Trauma survivorship has grown rapidly while defining the factors that influence post-traumatic quality of life has lagged. This study sought to develop a valid and reliable trauma-specific quality of life measure to help guide future post-traumatic research and clinical care. Methods Qualitative data were collected from adult trauma patients and their caregivers (Phase 1). Subsequent analysis of these data resulted in the development of a 59-item quality of life (QoL) questionnaire. The 59-item trauma-specific QoL questionnaire (T-QoL) was then administered to adult trauma patients (n=394) and a factor analysis was conducted. The validity of the final TQoL measurement tool was assessed (n=111) using the Medical Outcomes Study 36-Item Short Form Health Survey version 2 and the PTSD Checklist-Civilian Version (Phase 2). Results A 5-component structure using 43 items appeared to best represent the data. The 5 components included: Emotional Well-Being, Functional Engagement, Recovery/Resilience, Peri-Traumatic Experience, Physical Well-Being. Four of the five components were found to have strong Cronbach's alpha scores (>0.7), demonstrating consistent inter-item reliability. All subscales of the T-QoL correlated negatively with the PCL-C (p<.01), demonstrating that as the T-QoL increases, the likelihood of PTSD decreases. The physical well-being subscale of the T-QoL correlated significantly with the SF-36v2 PCS as did the emotional well-being subscale with the SF-36v2 MCS (p<.05). Conclusions This study utilized the experiences of trauma victims and their informal caregivers to develop a five-component, 43-item questionnaire with domains that are unique to trauma populations. Its accuracy and validity was confirmed using the PCL-C and the SF-36v2. We believe that the TQoL represents a novel

  9. Unconscious Semantic Activation Depends on Feature-Specific Attention Allocation

    ERIC Educational Resources Information Center

    Spruyt, Adriaan; De Houwer, Jan; Everaert, Tom; Hermans, Dirk

    2012-01-01

    We examined whether semantic activation by subliminally presented stimuli is dependent upon the extent to which participants assign attention to specific semantic stimulus features and stimulus dimensions. Participants pronounced visible target words that were preceded by briefly presented, masked prime words. Both affective and non-affective…

  10. Internet Content-Based Activities for English for Specific Purposes.

    ERIC Educational Resources Information Center

    Marco, Maria Jose Luzon

    2002-01-01

    Describes the benefits of a content-based approach to English for specific purposes, (ESP) examines how WebQuests can be integrated into a content-based ESP syllabus, analyze the different types of WebQuests suitable for ESP teaching, and discuses the limitations of this type of activity and the factors that should be considered when using…

  11. Essentials of specifications for activated alumina in defluoridation technology.

    PubMed

    Misra, S K

    2006-10-01

    Worldwide, fluoride occurs naturally in some aquifers at concentrations above the WHO guideline values or Bureau of Indian Standards or CPHEEO - MUD - GOI parametric values. Fluoride in excess of the permissible limits in drinking water causes a number of endemic conditions referred to collectively as "fluorosis". Endemic fluorosis remains a challenging national health problem in India and Rajasthan is one of the worst affected states in India though a wide range of chemical and physical defluoridation systems were evolved and tried. Still activated alumina is one of the most widely used and liked defluoridation material currently available. Boom in the advanced and versatile alumina technology has opened new vistas to avail the strong potential of selective alumina adsorbents which are application-specific. Non-regenerable and specialty alumina offer tremendous scope to defluoridate drinking water. Indian industries are manufacturing regenerable activated alumina for defluoridation of drinking water. In order to ensure application of an adsorbent, which caters the desired results with minimum interferences, health risks and long service life span, it is inevitable to draw out dimensions which define precisely the attributes of activated alumina. Specifications for activated alumina intended for defluoridation of drinking water, specific operating and performance requirements, and limitations expressed by critical analysis of cardinal characteristics pave way for adoption of acceptable specifications and code of practice at national level. PMID:18179116

  12. Measuring fluctuations in shear stretched DNAs using site specific labeling

    NASA Astrophysics Data System (ADS)

    Price, Allen; Graham, Thomas; Loparo, Joseph; Eaves, Joel

    2012-02-01

    We report a new technique for measuring the internal dynamics of surface tethered DNAs in shear flow. Previous studies have used end labeling or intercolating dyes which label the entire length of the DNA. Neither prior method can resolve the internal longitudinal fluctuations of the DNA. Our technique accomplishes this by site specific labeling of five sites in lambda phage DNA using EcoRI labeled with fluorescent quantum dots. We used our technique to determine the two point cross correlation functions of the longitudinal and transverse fluctuations of the DNA under shear flow. Our technique allows us to test current models of the non-equilibrium fluctuations of DNA in shear flow in a way previously inaccessible.

  13. Mechanisms of specificity in neuronal activity-regulated gene transcription

    PubMed Central

    Lyons, Michelle R.; West, Anne E.

    2011-01-01

    The brain is a highly adaptable organ that is capable of converting sensory information into changes in neuronal function. This plasticity allows behavior to be accommodated to the environment, providing an important evolutionary advantage. Neurons convert environmental stimuli into long-lasting changes in their physiology in part through the synaptic activity-regulated transcription of new gene products. Since the neurotransmitter-dependent regulation of Fos transcription was first discovered nearly 25 years ago, a wealth of studies have enriched our understanding of the molecular pathways that mediate activity-regulated changes in gene transcription. These findings show that a broad range of signaling pathways and transcriptional regulators can be engaged by neuronal activity to sculpt complex programs of stimulus-regulated gene transcription. However, the shear scope of the transcriptional pathways engaged by neuronal activity raises the question of how specificity in the nature of the transcriptional response is achieved in order to encode physiologically relevant responses to divergent stimuli. Here we summarize the general paradigms by which neuronal activity regulates transcription while focusing on the molecular mechanisms that confer differential stimulus-, cell-type-, and developmental-specificity upon activity-regulated programs of neuronal gene transcription. In addition, we preview some of the new technologies that will advance our future understanding of the mechanisms and consequences of activity-regulated gene transcription in the brain. PMID:21620929

  14. Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs.

    PubMed

    Keane, Fiona M; Yao, Tsun-Wen; Seelk, Stefanie; Gall, Margaret G; Chowdhury, Sumaiya; Poplawski, Sarah E; Lai, Jack H; Li, Youhua; Wu, Wengen; Farrell, Penny; Vieira de Ribeiro, Ana Julia; Osborne, Brenna; Yu, Denise M T; Seth, Devanshi; Rahman, Khairunnessa; Haber, Paul; Topaloglu, A Kemal; Wang, Chuanmin; Thomson, Sally; Hennessy, Annemarie; Prins, John; Twigg, Stephen M; McLennan, Susan V; McCaughan, Geoffrey W; Bachovchin, William W; Gorrell, Mark D

    2013-01-01

    The protease fibroblast activation protein (FAP) is a specific marker of activated mesenchymal cells in tumour stroma and fibrotic liver. A specific, reliable FAP enzyme assay has been lacking. FAP's unique and restricted cleavage of the post proline bond was exploited to generate a new specific substrate to quantify FAP enzyme activity. This sensitive assay detected no FAP activity in any tissue or fluid of FAP gene knockout mice, thus confirming assay specificity. Circulating FAP activity was ∼20- and 1.3-fold less in baboon than in mouse and human plasma, respectively. Serum and plasma contained comparable FAP activity. In mice, the highest levels of FAP activity were in uterus, pancreas, submaxillary gland and skin, whereas the lowest levels were in brain, prostate, leukocytes and testis. Baboon organs high in FAP activity included skin, epididymis, bladder, colon, adipose tissue, nerve and tongue. FAP activity was greatly elevated in tumours and associated lymph nodes and in fungal-infected skin of unhealthy baboons. FAP activity was 14- to 18-fold greater in cirrhotic than in non-diseased human liver, and circulating FAP activity was almost doubled in alcoholic cirrhosis. Parallel DPP4 measurements concorded with the literature, except for the novel finding of high DPP4 activity in bile. The new FAP enzyme assay is the first to be thoroughly characterised and shows that FAP activity is measurable in most organs and at high levels in some. This new assay is a robust tool for specific quantitation of FAP enzyme activity in both preclinical and clinical samples, particularly liver fibrosis. PMID:24371721

  15. Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs☆

    PubMed Central

    Keane, Fiona M.; Yao, Tsun-Wen; Seelk, Stefanie; Gall, Margaret G.; Chowdhury, Sumaiya; Poplawski, Sarah E.; Lai, Jack H.; Li, Youhua; Wu, Wengen; Farrell, Penny; Vieira de Ribeiro, Ana Julia; Osborne, Brenna; Yu, Denise M.T.; Seth, Devanshi; Rahman, Khairunnessa; Haber, Paul; Topaloglu, A. Kemal; Wang, Chuanmin; Thomson, Sally; Hennessy, Annemarie; Prins, John; Twigg, Stephen M.; McLennan, Susan V.; McCaughan, Geoffrey W.; Bachovchin, William W.; Gorrell, Mark D.

    2013-01-01

    The protease fibroblast activation protein (FAP) is a specific marker of activated mesenchymal cells in tumour stroma and fibrotic liver. A specific, reliable FAP enzyme assay has been lacking. FAP's unique and restricted cleavage of the post proline bond was exploited to generate a new specific substrate to quantify FAP enzyme activity. This sensitive assay detected no FAP activity in any tissue or fluid of FAP gene knockout mice, thus confirming assay specificity. Circulating FAP activity was ∼20- and 1.3-fold less in baboon than in mouse and human plasma, respectively. Serum and plasma contained comparable FAP activity. In mice, the highest levels of FAP activity were in uterus, pancreas, submaxillary gland and skin, whereas the lowest levels were in brain, prostate, leukocytes and testis. Baboon organs high in FAP activity included skin, epididymis, bladder, colon, adipose tissue, nerve and tongue. FAP activity was greatly elevated in tumours and associated lymph nodes and in fungal-infected skin of unhealthy baboons. FAP activity was 14- to 18-fold greater in cirrhotic than in non-diseased human liver, and circulating FAP activity was almost doubled in alcoholic cirrhosis. Parallel DPP4 measurements concorded with the literature, except for the novel finding of high DPP4 activity in bile. The new FAP enzyme assay is the first to be thoroughly characterised and shows that FAP activity is measurable in most organs and at high levels in some. This new assay is a robust tool for specific quantitation of FAP enzyme activity in both preclinical and clinical samples, particularly liver fibrosis. PMID:24371721

  16. A specific measurement matrix in compressive imaging system

    NASA Astrophysics Data System (ADS)

    Wang, Fen; Wei, Ping; Ke, Jun

    2011-11-01

    Compressed sensing or compressive sampling (CS) is a new framework for simultaneous data sampling and compression which was proposed by Candes, Donoho, and Tao several years ago. Ever since the advent of a single-pixel camera, one of the CS applications - compressive imaging (CI, also referred as feature-specific imaging) has aroused more interest of numerous researchers. However, it is still a challenging problem to choose a simple and efficient measurement matrix in such a hardware system, especially for large scale image. In this paper, we propose a new measurement matrix whose rows are the odd rows of N order Hadamard matrix and discuss the validity of the matrix theoretically. The advantage of the matrix is its universality and easy implementation in the optical domain owing to its integer-valued elements. In addition, we demonstrate the validity of the matrix through the reconstruction of natural images using Orthogonal Matching Pursuit (OMP) algorithm. Due to the limitation of the memory of the hardware system and personal computer which is used to simulate the process, it is impossible to create such a large matrix that is used to conduct large scale images. In order to solve the problem, the block-wise notion is introduced to conduct large scale images and the experiments results present the validity of this method.

  17. Metallic Magnetic Calorimeters for Absolute Activity Measurement

    NASA Astrophysics Data System (ADS)

    Loidl, M.; Leblanc, E.; Rodrigues, M.; Bouchard, J.; Censier, B.; Branger, T.; Lacour, D.

    2008-05-01

    We present a prototype of metallic magnetic calorimeters that we are developing for absolute activity measurements of low energy emitting radionuclides. We give a detailed description of the realization of the prototype, containing an 55Fe source inside the detector absorber. We present the analysis of first data taken with this detector and compare the result of activity measurement with liquid scintillation counting. We also propose some ways for reducing the uncertainty on the activity determination with this new technique.

  18. Stress causes tissue-specific changes in the sialyltransferase activity.

    PubMed

    Dabelic, Sanja; Flögel, Mirna; Maravić, Gordana; Lauc, Gordan

    2004-01-01

    Numerous pathological conditions are associated with specific changes in glycosylation. Recent studies clearly demonstrated a link between stress and the development and course of many diseases. Biochemical mechanisms that link stress and diseases are still not fully understood, but there are some indications that changes in glycosylation are involved in this process. Influence of acute and chronic psychological stress on protein sialylation as well as the activity of sialyltransferases, enzymes that synthesize sialoglycoproteins, has been studied on Fischer rats. Liver, spleen, kidney, skeletal muscle, heart, adrenal gland, serum, cerebellum, hippocampus, medulla oblongata and cortex have been analyzed. Statistically significant tissue- and type of stress-specific changes in total sialyltransferase (ST) activity were observed. Acute stress resulted in 39% increase of ST activity in liver and spleen, while at the same time there was 43% decrease in ST activity in cerebellum. In chronic stress, ST activity increased in spleen (93%) and decreased in liver (17%), cerebellum (38%) and hippocampus (64%). Western-blot analysis using Maackia amurensis and Sambucus nigra lectins did not reveal any difference in protein sialylation. The results of serum corticosterone analysis indicate that showed increase in acute stress and decrease in chronic stress are in good accordance with the hypothesis that corticosterone has a role in the regulation of liver ST activity. PMID:15241940

  19. Youth Physical Activity Resource Use and Activity Measured by Accelerometry

    ERIC Educational Resources Information Center

    Maslow, Andra L.; Colabianchi, Natalie

    2011-01-01

    Objectives: To examine whether use of physical activity resources (e.g., parks) was associated with daily physical activity measured by accelerometry. Methods: One hundred eleven adolescents completed a travel diary with concurrent accelerometry. The main exposure was self-reported use of a physical activity resource (none /1 resources). The main…

  20. Chemically sulfated natural galactomannans with specific antiviral and anticoagulant activities.

    PubMed

    Muschin, Tegshi; Budragchaa, Davaanyam; Kanamoto, Taisei; Nakashima, Hideki; Ichiyama, Koji; Yamamoto, Naoki; Shuqin, Han; Yoshida, Takashi

    2016-08-01

    Naturally occurring galactomannans were sulfated to give sulfated galactomannans with degrees of substitution of 0.7-1.4 per sugar unit and molecular weights of M¯n=0.6×10(4)-2.4×10(4). Sulfated galactomannans were found to have specific biological activities in vitro such as anticoagulant, anti-HIV and anti-Dengue virus activities. The biological activities were compared with those of standard dextran and curdlan sulfates, which are polysaccharides with potent antiviral activity and low cytotoxicity. It was found that sulfated galactomannans had moderate to high anticoagulant activity, 13.4-36.6unit/mg, compared to that of dextran and curdlan sulfates, 22.7 and 10.0unit/mg, and high anti-HIV and anti-Dengue virus activities, 0.04-0.8μg/mL and 0.2-1.1μg/mL, compared to those curdlan sulfates, 0.1μg/mL, respectively. The cytotoxicity on MT-4 and LCC-MK2 cells was low. Surface plasmon resonance (SPR) of sulfated galactomannans revealed strong interaction with poly-l-lysine as a model compound of virus proteins, and suggested that the specific biological activities might originate in the electrostatic interaction of negatively charged sulfate groups of sulfated galactomannans and positively charged amino groups of surface proteins of viruses. These results suggest that sulfated galactomannans effectively prevented the infection of cells by viruses and the degree of substitution and molecular weights played important roles in the biological activities. PMID:27154517

  1. Novel strategies for ultrahigh specific activity targeted nanoparticles

    SciTech Connect

    Zhou, Dong

    2012-12-13

    We have developed novel strategies optimized for preparing high specific activity radiolabeled nanoparticles, targeting nuclear imaging of low abundance biomarkers. Several compounds have been labeled with F-18 and Cu-64 for radiolabeling of SCK-nanoparticles via Copper(I) catalyzed or copper-free alkyne-azide cyclolization. Novel strategies have been developed to achieve ultrahigh specific activity with administrable amount of dose for human study using copper-free chemistry. Ligands for carbonic anhydrase 12 (CA12), a low abundance extracellular biomarker for the responsiveness of breast cancer to endocrine therapie, have been labeled with F-18 and Cu-64, and one of them has been evaluated in animal models. The results of this project will lead to major improvements in the use of nanoparticles in nuclear imaging and will significantly advance their potential for detecting low abundance biomarkers of medical importance.

  2. Method of preparing high specific activity platinum-195m

    DOEpatents

    Mirzadeh, Saed; Du, Miting; Beets, Arnold L.; Knapp, Jr., Furn F.

    2004-06-15

    A method of preparing high-specific-activity .sup.195m Pt includes the steps of: exposing .sup.193 Ir to a flux of neutrons sufficient to convert a portion of the .sup.193 Ir to .sup.195m Pt to form an irradiated material; dissolving the irradiated material to form an intermediate solution comprising Ir and Pt; and separating the Pt from the Ir by cation exchange chromatography to produce .sup.195m Pt.

  3. Method for preparing high specific activity 177Lu

    SciTech Connect

    Mirzadeh, Saed; Du, Miting; Beets, Arnold L.; Knapp, Jr., Furn F.

    2004-04-06

    A method of separating lutetium from a solution containing Lu and Yb, particularly reactor-produced .sup.177 Lu and .sup.177 Yb, includes the steps of: providing a chromatographic separation apparatus containing LN resin; loading the apparatus with a solution containing Lu and Yb; and eluting the apparatus to chromatographically separate the Lu and the Yb in order to produce high-specific-activity .sup.177 Yb.

  4. Measuring layer-specific depth-of-focus requirements

    NASA Astrophysics Data System (ADS)

    Liegl, Bernhard; Gabor, Allen; Brodsky, Colin; Cotte, John; Krishnan, Mahadevaiyer

    2008-03-01

    As the Rayleigh equations already tell us, improvements in imaging resolution often come at the price of a depth-offocus loss. Often we balance the resolution versus DoF dilemma without regard of the imaging layers location in the overall film stack. E.g. often several via or metal layers are processed with the same optical settings despite facing different amount of depth-of-focus requirements. In actuality, however, substrate induced focus variation can vary greatly from layers at the bottom of a film stack to the layers higher up in the film stack. In the age of super-low k1 lithography this variance needs to be taken into account on a layer specific basis when evaluating the resolution versus DoF tradeoff. We have studied substrate induced focus variation for a 45nm technology test-site as function of film stack sequence and spatial frequency, combining various measurement techniques into an overall topography spectrum. These techniques include data extraction from the exposure tools optical leveling sensor, a mechanical air gauge to calibrate the former and interferometric profiling tools. As a result, we can quantify our DoF requirement for a given layer and product and use this information to optimize our process design on a layer-by-layer basis. This work was performed by the Research Alliance Teams at various IBM Research and Development Facilities

  5. Measurement of bone specific alkaline phosphatase in the horse: a comparison of two techniques.

    PubMed

    Jackson, B; Eastell, R; Russell, R G; Lanyon, L E; Price, J S

    1996-09-01

    For many years total alkaline phosphatase (AP) activity in serum has been used to monitor bone metabolism in different species. However, total AP lacks bone specificity because the total activity in serum is made up of several isoenzymes, of which the liver and bone isoforms predominate. The aim of the present study was to evaluate an immunoradiometric assay for measuring bone specific alkaline phosphatase (BAP) in horses. BAP, a specific marker of bone formation, was measured in sera from thoroughbred horses by using a previously characterised wheat germ lectin (WGL) precipitation assay and an immunoradiometric assay. The levels of immunoreactive BAP (iBAP) and WGL precipitated BAP (wBAP) were related to the serum levels of total AP and another marker of bone formation, the carboxy-terminal propeptide of type 1 collagen (PICP). In horses over one year old, iBAP correlated at least as strongly with total AP as with wBAP, which suggests that the immunoradiometric assay may partially cross-react with liver alkaline phosphatase in horse serum. This possibility was supported by the observation that there was a weaker correlation between iBAP and PICP than between wBAP and PICP. These data indicate that WGL precipitation is currently the most specific method for measuring bone specific alkaline phosphatase in horses. PMID:8880988

  6. Measurement of Larval Activity in the Drosophila Activity Monitor

    PubMed Central

    McParland, Aidan L.; Follansbee, Taylor L.; Ganter, Geoffrey K.

    2016-01-01

    Drosophila larvae are used in many behavioral studies, yet a simple device for measuring basic parameters of larval activity has not been available. This protocol repurposes an instrument often used to measure adult activity, the TriKinetics Drosophila activity monitor (MB5 Multi-Beam Activity Monitor) to study larval activity. The instrument can monitor the movements of animals in 16 individual 8 cm glass assay tubes, using 17 infrared detection beams per tube. Logging software automatically saves data to a computer, recording parameters such as number of moves, times sensors were triggered, and animals’ positions within the tubes. The data can then be analyzed to represent overall locomotion and/or position preference as well as other measurements. All data are easily accessible and compatible with basic graphing and data manipulation software. This protocol will discuss how to use the apparatus, how to operate the software and how to run a larval activity assay from start to finish. PMID:25993121

  7. Cell-specific activity of neprilysin 2 isoforms and enzymic specificity compared with neprilysin.

    PubMed Central

    Rose, Christiane; Voisin, Stéphanie; Gros, Claude; Schwartz, Jean-Charles; Ouimet, Tanja

    2002-01-01

    Neprilysin (NEP) 2 is a recently cloned glycoprotein displaying a high degree of sequence identity with neprilysin (EC 3.4.24.11), the prototypical member of the M13 subfamily of metalloproteases. Whereas NEP is involved in the metabolism of several bioactive peptides by plasma membranes of various cells, the enzymic properties and physiological functions of NEP2 are unknown. Here we characterize the cell-expression modalities and enzymic specificity of two alternatively spliced isoforms of NEP2 in Chinese hamster ovary and AtT20 cells. In the two cell lines, both isoforms are type II glycoproteins inserted in the endoplasmic reticulum as inactive precursors. Maturation detected by Western-blot analysis of glycosidase digests was cell-specific and more efficient in the endocrine cell line. The enzymic activity of both isoforms semi-purified from AtT20 cells reveals comparable specificities in terms of model substrates, pH optima and inhibitory patterns. NEP2 activity was compared with that of NEP regarding potencies of transition-state inhibitors, modes of hydrolysis, maximal hydrolysis rates and apparent affinities of bioactive peptides. Although all transition-state inhibitors of NEP inhibited NEP2 activity, albeit with different potencies, and many peptides were cleaved at the same amide bond by both peptidases, differences could be observed, i.e. in the hydrolysis of gonadotropin-releasing hormone and cholecystokinin, which occurred at different sites and more efficiently in the case of NEP2. Differences in cleavage of bioactive peptides, in cell-trafficking patterns and in tissue distribution indicate that NEP and NEP2 play distinct physiological roles in spite of their high degree of sequence identity. PMID:11964170

  8. Measuring Homework Completion in Behavioral Activation

    ERIC Educational Resources Information Center

    Busch, Andrew M.; Uebelacker, Lisa A.; Kalibatseva, Zornitsa; Miller, Ivan W.

    2010-01-01

    The aim of this study was to develop and validate an observer-based coding system for the characterization and completion of homework assignments during Behavioral Activation (BA). Existing measures of homework completion are generally unsophisticated, and there is no current measure of homework completion designed to capture the particularities…

  9. Impact of dewatering technologies on specific methanogenic activity.

    PubMed

    Batstone, Damien J; Lu, Yang; Jensen, Paul D

    2015-10-01

    Dewatering methods for recuperative thickening and final dewatering can potentially impact methanogenic activity and microbial community. This influences both the feasibility of recuperative thickening to increase solids residence time within a digester, and the utilisation of dewatered digestate as inoculum for new digesters. Thickening technology can reduce methanogenic activity through either air contact (rotary drum, DAF, or belt filter press), or by lysing cells through shear (centrifuge). To assess this, two plants with recuperative thickening (rotary drum) in their anaerobic digester, and five without recuperative thickening, had specific methanogenic activity tested in all related streams, including dewatering feed, thickened return, final cake, and centrate. All plants had high speed centrifuges for final dewatering. The digester microbial community was also assessed through 16s pyrotag sequencing and subsequent principal component analysis (PCA). The specific methanogenic activity of all samples was in the expected range of 0.2-0.4 gCOD gVS(-1)d(-1). Plants with recuperative thickening did not have lower digester activity. Centrifuge based dewatering had a significant and variable impact on methanogenic activity in all samples, ranging between 20% and 90% decrease but averaging 54%. Rotary drum based recuperative thickening had a far smaller impact on activity, with a 0% per-pass drop in activity in one plant, and a 20% drop in another. However, the presence of recuperative thickening was a major predictor of overall microbial community (PC1, p = 0.0024). Microbial community PC3 (mainly driven by a shift in methanogens) was a strong predictor for sensitivity in activity to shear (p = 0.0005, p = 0.00001 without outlier). The one outlier was related to a plant producing the wettest cake (17% solids). This indicates that high solids is a potential driver of sensitivity to shear, but that a resilient microbial community can also bestow resilience

  10. Self-Specific Stimuli Interact Differently than Non-Self-Specific Stimuli with Eyes-Open Versus Eyes-Closed Spontaneous Activity in Auditory Cortex

    PubMed Central

    Qin, Pengmin; Grimm, Simone; Duncan, Niall W.; Holland, Giles; Guo, Jia shen; Fan, Yan; Weigand, Anne; Baudewig, Juergen; Bajbouj, Malek; Northoff, Georg

    2013-01-01

    Previous studies suggest that there may be a distinct relationship between spontaneous neural activity and subsequent or concurrent self-specific stimulus-induced activity. This study aims to test the impact of spontaneous activity as recorded in an eyes-open (EO) resting state as opposed to eyes-closed (EC) on self-specific versus non-self-specific auditory stimulus-induced activity in fMRI. In our first experiment we used self-specific stimuli comprised of the subject’s own name and non-self-specific stimuli comprised of a friend’s name and an unknown name, presented during EO versus EC baselines in a 3 name condition × 2 baseline design. In Experiment 2 we directly measured spontaneous activity in the absence of stimuli during EO versus EC to confirm a modulatory effect of the two baseline conditions in the regions found to show an interaction effect in Experiment 1. Spontaneous activity during EO was significantly higher than during EC in bilateral auditory cortex and non-self-specific names yielded stronger signal changes relative to EO baseline than to EC. In contrast, there was no difference in response to self-specific names relative to EO baseline than to EC despite the difference between spontaneous activity levels. These results support an impact of spontaneous activity on stimulus-induced activity, moreover an impact that depends on the high-level stimulus characteristic of self-specificity. PMID:23908625

  11. 40 CFR 90.416 - Intake air flow measurement specifications.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... percent of full-scale value of the measurement device for all modes except the idle mode. For the idle mode, the measurement accuracy must be ±five percent or less of the full-scale value. The...

  12. SITE-SPECIFIC MEASUREMENTS OF RESIDENTIAL RADON PROTECTION CATEGORY

    EPA Science Inventory

    The report describes a series of benchmark measurements of soil radon potential at seven Florida sites and compares the measurements with regional estimates of radon potential from the Florida radon protection map. The measurements and map were developed under the Florida Radon R...

  13. Cyclotron production of ``very high specific activity'' platinum radiotracers in No Carrier Added form

    NASA Astrophysics Data System (ADS)

    Birattari, C.; Bonardi, M.; Groppi, F.; Gini, L.; Gallorini, M.; Sabbioni, E.; Stroosnijder, M. F.

    2001-12-01

    At the "Radiochemistry Laboratory" of Accelerators and Applied Superconductivity Laboratory, LASA, several production and quality assurance methods for short-lived and high specific activity radionuclides, have been developed. Presently, the irradiations are carried out at the Scanditronix MC40 cyclotron (K=38; p, d, He-4 and He-3) of JRC-Ispra, Italy, of the European Community, while both chemical purity and specific activity determination are carried out at the TRIGA MARK II research reactor of University of Pavia and at LASA itself. In order to optimize the irradiation conditions for platinum radiotracer production, both thin- and thick-target excitation function of natOs(α,xn) nuclear reactions were measured. A very selective radiochemical separation to obtain Pt radiotracers in No Carrier Added form, has been developed. Both real specific activity and chemical purity of radiotracer, have been determined by neutron activation analysis and atomic absorption spectrometry. An Isotopic Dilution Factor (IDF) of the order of 50 is achieved.

  14. Detection of Telomerase Activity Using Capacitance Measurements

    NASA Astrophysics Data System (ADS)

    Kang, Bong Keun; Lee, Ri Mi; Choi, Ahmi; Jung, Hyo-Il; Yoo, Kyung-Hwa

    2007-03-01

    Telomerase activity has been found in about 85% cancer cells, while no activity observed in normal cells, so that telomerase has been proposed as a marker for cancer detection. Here, we describe electrical detection of telomerase activity using capacitance measurements. We have investigated the length dependence of capacitance on DNA solutions and found that the capacitance of DNA solutions were dependent on the DNA length. In addition, upon adding telomerase into the solution of telomeric substrate primer, the capacitance was observed to change as a function of time due to the telomeric elongation. These results suggest that this novel nanosensor may be used for rapid detection of telomerase activity.

  15. Measuring the Built Environment for Physical Activity

    PubMed Central

    Brownson, Ross C.; Hoehner, Christine M.; Day, Kristen; Forsyth, Ann; Sallis, James F.

    2009-01-01

    Physical inactivity is one of the most important public health issues in the U.S. and internationally. Increasingly, links are being identified between various elements of the physical—or built—environment and physical activity. To understand the impact of the built environment on physical activity, the development of high-quality measures is essential. Three categories of built environment data are being used: (1) perceived measures obtained by telephone interview or self-administered questionnaires; (2) observational measures obtained using systematic observational methods (audits); and (3) archival data sets that are often layered and analyzed with GIS. This review provides a critical assessment of these three types of built-environment measures relevant to the study of physical activity. Among perceived measures, 19 questionnaires were reviewed, ranging in length from 7 to 68 questions. Twenty audit tools were reviewed that cover community environments (i.e., neighborhoods, cities), parks, and trails. For GIS-derived measures, more than 50 studies were reviewed. A large degree of variability was found in the operationalization of common GIS measures, which include population density, land-use mix, access to recreational facilities, and street pattern. This first comprehensive examination of built-environment measures demonstrates considerable progress over the past decade, showing diverse environmental variables available that use multiple modes of assessment. Most can be considered first-generation measures, so further development is needed. In particular, further research is needed to improve the technical quality of measures, understand the relevance to various population groups, and understand the utility of measures for science and public health. PMID:19285216

  16. Sensitivity and Specificity of Hypnosis Effects on Gastric Myoelectrical Activity

    PubMed Central

    Enck, Paul; Weimer, Katja; Muth, Eric R.; Zipfel, Stephan; Martens, Ute

    2013-01-01

    Objectives The effects of hypnosis on physiological (gastrointestinal) functions are incompletely understood, and it is unknown whether they are hypnosis-specific and gut-specific, or simply unspecific effects of relaxation. Design Sixty-two healthy female volunteers were randomly assigned to either a single session of hypnotic suggestion of ingesting an appetizing meal and an unappetizing meal, or to relax and concentrate on having an appetizing or unappetizing meal, while the electrogastrogram (EGG) was recorded. At the end of the session, participants drank water until they felt full, in order to detect EGG-signal changes after ingestion of a true gastric load. During both conditions participants reported their subjective well-being, hunger and disgust at several time points. Results Imagining eating food induced subjective feelings of hunger and disgust as well as changes in the EGG similar to, but more pronounced than those seen with a real gastric water load during both hypnosis and relaxation conditions. These effects were more pronounced when imagining an appetizing meal than with an unappetizing meal. There was no significant difference between the hypnosis and relaxation conditions. Conclusion Imagination with and without hypnosis exhibits similar changes in subjective and objective measures in response to imagining an appetizing and an unappetizing food, indicating high sensitivity but low specificity. PMID:24358287

  17. Selecting Level-Specific Specialized Vocabulary Using Statistical Measures

    ERIC Educational Resources Information Center

    Chujo, Kiyomi; Utiyama, Masao

    2006-01-01

    To find an easy-to-use, automated tool to identify technical vocabulary applicable to learners at various levels, nine statistical measures were applied to the 7.3-million-word "commerce and finance" component of the British National Corpus. The resulting word lists showed that each statistical measure extracted a different level of specialized…

  18. Differential Reovirus-Specific and Herpesvirus-Specific Activator Protein 1 Activation of Secretogranin II Leads to Altered Virus Secretion

    PubMed Central

    Berard, Alicia R.; Severini, Alberto

    2015-01-01

    ABSTRACT Viruses utilize host cell machinery for propagation and manage to evade cellular host defense mechanisms in the process. Much remains unknown regarding how the host responds to viral infection. We recently performed global proteomic screens of mammalian reovirus TIL- and T3D-infected and herpesvirus (herpes simplex virus 1 [HSV-1])-infected HEK293 cells. The nonenveloped RNA reoviruses caused an upregulation, whereas the enveloped DNA HSV-1 caused a downregulation, of cellular secretogranin II (SCG2). SCG2, a member of the granin family that functions in hormonal peptide sorting into secretory vesicles, has not been linked to virus infections previously. We confirmed SCG2 upregulation and found SCG2 phosphorylation by 18 h postinfection (hpi) in reovirus-infected cells. We also found a decrease in the amount of reovirus secretion from SCG2 knockdown cells. Similar analyses of cells infected with HSV-1 showed an increase in the amount of secreted virus. Analysis of the stress-activated protein kinase (SAPK)/Jun N-terminal protein kinase (JNK) pathway indicated that each virus activates different pathways leading to activator protein 1 (AP-1) activation, which is the known SCG2 transcription activator. We conclude from these experiments that the negative correlation between SCG2 quantity and virus secretion for both viruses indicates a virus-specific role for SCG2 during infection. IMPORTANCE Mammalian reoviruses affect the gastrointestinal system or cause respiratory infections in humans. Recent work has shown that all mammalian reovirus strains (most specifically T3D) may be useful oncolytic agents. The ubiquitous herpes simplex viruses cause common sores in mucosal areas of their host and have coevolved with hosts over many years. Both of these virus species are prototypical representatives of their viral families, and investigation of these viruses can lead to further knowledge of how they and the other more pathogenic members of their respective

  19. Specific Remedy for Specific Problem: Measuring Service Quality in South African Higher Education

    ERIC Educational Resources Information Center

    de Jager, Johan; Gbadamosi, Gbolahan

    2010-01-01

    This study commences a process of developing a scale for the measurement of service quality in higher education in South Africa and also examines the relationship between the measures of service quality on the one hand and some other related variables such as intention to leave the university, trust in management of the university and the overall…

  20. 40 CFR 91.416 - Intake air flow measurement specifications.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... operating range during the test. Overall measurement accuracy must be ±2 percent of full-scale value of the... percent or less of the full-scale value. The Administrator must be advised of the method used prior...

  1. 40 CFR 86.314-79 - Fuel flow measurement specifications.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... volume flow and density, the error in the actual volume consumed must not be greater than ±1 percent of... (electronic weight, volume, density, etc.), measurements may not be used for calculations if the...

  2. Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures; January 2012 - March 2013

    SciTech Connect

    Jayaweera, T.; Haeri, H.

    2013-04-01

    Under the Uniform Methods Project, DOE is developing a framework and a set of protocols for determining the energy savings from specific energy efficiency measures and programs. The protocols provide a straightforward method for evaluating gross energy savings for common residential and commercial measures offered in ratepayer-funded initiatives in the United States. They represent a refinement of the body of knowledge supporting energy efficiency evaluation, measurement, and verification (EM&V) activities. This document deals with savings from the following measures: commercial and industrial lighting, commercial and industrial lighting controls, small commercial and residential unitary and split system HVAC cooling equipment, residential furnaces and boilers, residential lighting, refrigerator recycling, whole-building retrofit using billing analysis, metering, peak demand and time-differentiated energy savings, sample design, survey design and implementation, and assessing persistence and other evaluation issues.

  3. Preparation and immunoreactivity of high specific activity indium-111-DTPA labeled monoclonal antibody (MoAb) using ultrapure indium-111

    SciTech Connect

    Zoghbi, S.S.; Neumann, R.D.; Gottschalk, A.

    1986-10-01

    The preparation of high-specific activity /sup 111/In-DTPA-MoAb without increasing the number of DTPA molecules per Ab was investigated. Instant thin layer chromatography was used to assay the relationship between labeling efficiencies and specific activities. With ultrapurified /sup 111/In, the specific activity of the radiolabeled MoAb approached the expected theoretic maximum of 100 muCi/microgram. The bioactivity of such high-specific activity preparation showed no degradation as measured by in vitro cell binding assay.

  4. Performance evaluation of quantitative adiabatic (13)C NMR pulse sequences for site-specific isotopic measurements.

    PubMed

    Thibaudeau, Christophe; Remaud, Gérald; Silvestre, Virginie; Akoka, Serge

    2010-07-01

    (2)H/(1)H and (13)C/(12)C site-specific isotope ratios determined by NMR spectroscopy may be used to discriminate pharmaceutically active ingredients based on the synthetic process used in production. Extending the Site-specific Natural Isotope Fractionation NMR (SNIF-NMR) method to (13)C is highly beneficial for complex organic molecules when measurements of (2)H/(1)H ratios lead to poorly defined molecular fingerprints. The current NMR methodology to determine (13)C/(12)C site-specific isotope ratios suffers from poor sensitivity and long experimental times. In this work, several NMR pulse sequences based on polarization transfer were evaluated and optimized to measure precise quantitative (13)C NMR spectra within a short time. Adiabatic 180 degrees (1)H and (13)C pulses were incorporated into distortionless enhancement by polarization transfer (DEPT) and refocused insensitive nuclei enhanced by polarization transfer (INEPT) to minimize the influence of 180 degrees pulse imperfections and of off-resonance effects on the precision of the measured (13)C peak areas. The adiabatic DEPT sequence was applied to draw up a precise site-specific (13)C isotope profile of ibuprofen. A modified heteronuclear cross-polarization (HCP) experiment featuring (1)H and (13)C spin-locks with adiabatic 180 degrees pulses is also introduced. This sequence enables efficient magnetization transfer across a wide (13)C frequency range although not enough for an application in quantitative (13)C isotopic analysis. PMID:20527737

  5. Measuring Active Learning to Predict Course Quality

    ERIC Educational Resources Information Center

    Taylor, John E.; Ku, Heng-Yu

    2011-01-01

    This study investigated whether active learning within computer-based training courses can be measured and whether it serves as a predictor of learner-perceived course quality. A major corporation participated in this research, providing access to internal employee training courses, training representatives, and historical course evaluation data.…

  6. Allele-specific DNA methylation reinforces PEAR1 enhancer activity.

    PubMed

    Izzi, Benedetta; Pistoni, Mariaelena; Cludts, Katrien; Akkor, Pinar; Lambrechts, Diether; Verfaillie, Catherine; Verhamme, Peter; Freson, Kathleen; Hoylaerts, Marc F

    2016-08-18

    Genetic variation in the PEAR1 locus is linked to platelet reactivity and cardiovascular disease. The major G allele of rs12041331, an intronic cytosine guanine dinucleotide-single-nucleotide polymorphism (CpG-SNP), is associated with higher PEAR1 expression in platelets and endothelial cells than the minor A allele. The molecular mechanism underlying this difference remains elusive. We have characterized the histone modification profiles of the intronic region surrounding rs12041331 and identified H3K4Me1 enhancer-specific enrichment for the region that covers the CpG-SNP. Interestingly, methylation studies revealed that the CpG site is fully methylated in leukocytes of GG carriers. Nuclear protein extracts from megakaryocytes, endothelial cells, vs control HEK-293 cells show a 3-fold higher affinity for the methylated G allele compared with nonmethylated G or A alleles in a gel electrophoretic mobility shift assay. To understand the positive relationship between methylation and gene expression, we studied DNA methylation at 4 different loci of PEAR1 during in vitro megakaryopoiesis. During differentiation, the CpG-SNP remained fully methylated, while we observed rapid methylation increases at the CpG-island overlapping the first 5'-untranslated region exon, paralleling the increased PEAR1 expression. In the same region, A-allele carriers of rs12041331 showed significantly lower DNA methylation at CGI1 compared with GG homozygote. This CpG-island contains binding sites for the methylation-sensitive transcription factor CTCF, whose binding is known to play a role in enhancer activation and/or repression. In conclusion, we report the molecular characterization of the first platelet function-related CpG-SNP, a genetic predisposition that reinforces PEAR1 enhancer activity through allele-specific DNA methylation. PMID:27313330

  7. Application-specific architectures of CMOS monolithic active pixel sensors

    NASA Astrophysics Data System (ADS)

    Szelezniak, Michal; Besson, Auguste; Claus, Gilles; Colledani, Claude; Degerli, Yavuz; Deptuch, Grzegorz; Deveaux, Michael; Dorokhov, Andrei; Dulinski, Wojciech; Fourches, Nicolas; Goffe, Mathieu; Grandjean, Damien; Guilloux, Fabrice; Heini, Sebastien; Himmi, Abdelkader; Hu, Christine; Jaaskelainen, Kimmo; Li, Yan; Lutz, Pierre; Orsini, Fabienne; Pellicioli, Michel; Shabetai, Alexandre; Valin, Isabelle; Winter, Marc

    2006-11-01

    Several development directions intended to adapt and optimize monolithic active pixel sensors for specific applications are presented in this work. The first example, compatible with the STAR microvertex upgrade, is based on a simple two-transistor pixel circuitry. It is suited for a long integration time, room-temperature operation and minimum power dissipation. In another approach for this application, a specific readout method is proposed, allowing optimization of the integration time independently of the full frame-readout time. The circuit consists of an in-pixel front-end voltage amplifier, with a gain on the order of five, followed by two analog memory cells. The extended version of this scheme, based on the implementation of more memory cells per pixel, is the solution considered for the outer layers of a microvertex detector at the international linear collider. For the two innermost layers, a circuit allowing fast frame scans together with on-line, on-chip data sparsification is proposed. The first results of this prototype demonstrate that the fixed pattern dispersion is reduced below a noise level of 15 e -, allowing the use of a single comparator or a low-resolution ADC per pixel column. A common element for most of the mentioned readout schemes is a low-noise, low power consumption, layout efficient in-pixel amplifier. A review of possible solutions for this element together with some experimental results is presented.

  8. Measurements of thermospheric response to auroral activities

    NASA Technical Reports Server (NTRS)

    Okano, S.; Kim, J. S.

    1986-01-01

    The Joule heating produced by auroral electrojets and its thermospheric response can be studied by monitoring the thermospheric temperatures by optical methods; simultaneously, the concurrent auroral electrojet activities can be investigated by using geomagnetic records obtained from stations along a meridian close to the observation site of optical measurements. The measurements are reported of thermospheric response to auroral activities which were made at Albany (42.68 deg N, 73.82 deg W), New York on September 2, 1978 (UT) when an isolated substorm occured. The thermospheric temperatures were measured by using a high resolution Fabry-Perot interferometer that determines the line profiles of the (OI) 6300A line emission. The intensities and latitudinal positions of auroral electrojets were obtained by the analysis of magnetograms from the IMS Fort Churchill meridian chain stations.

  9. Site-specific PEGylation of lidamycin and its antitumor activity.

    PubMed

    Li, Liang; Shang, Boyang; Hu, Lei; Shao, Rongguang; Zhen, Yongsu

    2015-05-01

    In this study, N-terminal site-specific mono-PEGylation of the recombinant lidamycin apoprotein (rLDP) of lidamycin (LDM) was prepared using a polyethyleneglycol (PEG) derivative (M w 20 kDa) through a reactive terminal aldehyde group under weak acidic conditions (pH 5.5). The biochemical properties of mPEG-rLDP-AE, an enediyne-integrated conjugate, were analyzed by SDS-PAGE, RP-HPLC, SEC-HPLC and MALDI-TOF. Meanwhile, in vitro and in vivo antitumor activity of mPEG-rLDP-AE was evaluated by MTT assays and in xenograft model. The results indicated that mPEG-rLDP-AE showed significant antitumor activity both in vitro and in vivo. After PEGylation, mPEG-rLDP still retained the binding capability to the enediyne AE and presented the physicochemical characteristics similar to that of native LDP. It is of interest that the PEGylation did not diminish the antitumor efficacy of LDM, implying the possibility that this derivative may function as a payload to deliver novel tumor-targeted drugs. PMID:26579455

  10. Measuring the Specific Heat of Metals by Cooling

    ERIC Educational Resources Information Center

    Dittrich, William; Minkin, Leonid; Shapovalov, Alexander S.

    2010-01-01

    Three in one? Yes, three standard undergraduate thermodynamics experiments in one, not an oval can of lubricating oil. Previously it has been shown that the PASCO scientific apparatus for measuring coefficients of thermal expansion of metals can also be used to illustrate Newton's law of cooling in the same experiment. Now it will be shown that by…

  11. Functional specification of the Performance Measurement (PM) module

    NASA Technical Reports Server (NTRS)

    Berliner, J. E.

    1980-01-01

    The design of the Performance Measurement Module is described with emphasis on what the PM Module would do, and what it would look like to the user. The PM Module as described could take several man-years to develop. An evolutionary approach to the implementation of the PM Module is presented which would provide an operational baseline PM Module within a few months.

  12. Assessing General and Specific Attitudes in Human Learning Behavior: An Activity Perspective and a Multilevel Modeling Approach

    ERIC Educational Resources Information Center

    Sun, Jun; Willson, Victor L.

    2008-01-01

    This article proposes a multilevel modeling approach to study the general and specific attitudes formed in human learning behavior. Based on the premises of activity theory, it conceptualizes the unit of analysis for attitude measurement as a scalable and evolving activity system rather than a single action. Measurement issues related to this…

  13. Task Specific Frequencies of Neck Motion Measured in Healthy Young Adults over a 5 Day Period

    PubMed Central

    Cobian, Daniel G.; Sterling, Andrew C.; Anderson, Paul A.; Heiderscheit, Bryan C.

    2010-01-01

    Study Design Observational cohort design. Objective To quantify the frequencies and magnitudes of neck motion during daily activities in healthy subjects. Summary of Background Data Previous studies have measured the maximum excursions during re-created ADLs in lab settings, but there is a lack of information available on frequencies and excursions of neck motion with ADLs in non-artificial settings. Methods Ten healthy young adults were fitted with a portable motion measurement device that recorded movement about each primary axis. Participants were instructed to wear the unit continuously over a 5-day period and record their daily activities with corresponding times. After the collection period, subjects' activity logs were analyzed and data were partitioned into five categories which provided the most primary representation of ADLs: athletics, work, travel, sleep, and miscellaneous. Each category was further divided into increasingly specific activities (e.g. running and walking). Frequency of motions within 5° increments was determined and an hourly rate was calculated for each activity. Median motion about each axis for each activity was also determined. Results The total number of movements per hour for all axes, regardless of amplitude, was highest during athletic activity and lowest during sleeping. The majority of movements (92% of athletic activity, 90% of work) required less than 25° of lateral bending, while greater range of movement requirements were observed for flexion-extension and axial rotation. The median range of motion along all axes was highest for athletic activity and lowest for sleeping. Conclusions The results of this study provide a baseline of the frequency and magnitude of neck motion during normal ADLs for the specified population. These findings can assist physicians and physical therapists in determining the extent of disability and identifying activities that will likely be problematic for patients with limited cervical motion

  14. Enhancer-specific modulation of E protein activity.

    PubMed

    Markus, Maurice; Du, Zhimei; Benezra, Robert

    2002-02-22

    Homodimeric complexes of members of the E protein family of basic helix-loop-helix (bHLH) transcription factors are important for tissue-specific activation of genes in B lymphocytes (Bain, G., Gruenwald, S., and Murre, C. (1993) Mol. Cell Biol. 13, 3522-3529; Shen, C. P., and Kadesch, T. (1995) Mol. Cell Biol. 15, 4518-4524; Jacobs, Y., et al. (1994) Mol. Cell Biol. 14, 4087-4096; Wilson, R. B., et al. (1991) Mol. Cell Biol. 11, 6185-6191). These homodimers, however, have little activity on myogenic enhancers (Weintraub, H., Genetta, T., and Kadesch, T. (1994) Genes Dev. 8, 2203-2211). We report here the identification of a novel cis-acting transcriptional repression domain in the E protein family of bHLH transcription factors. This domain, the Rep domain, is present in each of the known vertebrate E proteins. Extensive mapping analysis demonstrates that this domain is an acidic region of 30 amino acids with a predicted loop structure. Fusion studies indicate that the Rep domain can repress both of the E protein transactivation domains (AD1 and AD2). Physiologically, the Rep domain plays a key role in maintaining E protein homodimers in an inactive state on myogenic enhancers. In addition, we demonstrate that Rep domain mediated repression of AD1 is a necessary for the function of MyoD-E protein heterodimeric complexes. These studies demonstrate that the Rep domain is important for modulating the transcriptional activity of E proteins and provide key insights into both the selectivity and mechanism of action of E protein containing bHLH protein complexes. PMID:11724804

  15. Activity measurements of radon from construction materials.

    PubMed

    Fior, L; Nicolosi Corrêa, J; Paschuk, S A; Denyak, V V; Schelin, H R; Soreanu Pecequilo, B R; Kappke, J

    2012-07-01

    This work presents the results of radon concentration measurements of construction materials used in the Brazilian industry, such as clay (red) bricks and concrete blocks. The measurements focused on the detection of indoor radon activity during different construction stages and the analysis of radionuclides present in the construction materials. For this purpose, sealed chambers with internal dimensions of approximately 60×60×60 cm3 were built within a protected and isolated laboratory environment, and stable air humidity and temperature levels were maintained. These chambers were also used for radon emanation reduction tests. The chambers were built in four major stages: (1) assembly of the walls using clay (red) bricks, concrete blocks, and mortar; (2) installation of plaster; (3) finishing of wall surface using lime; and (4) insulation of wall surface and finishing using paint. Radon measurements were performed using polycarbonate etched track detectors. By comparing the three layers applied to the masonry walls, it was concluded that only the last step (wall painting using acrylic varnish) reduced the radon emanation, by a factor of approximately 2. Samples of the construction materials (clay bricks and concrete blocks) were ground, homogenized, and subjected to gamma-ray spectrometry analysis to evaluate the activity concentrations of 226Ra, 232Th and 40K. The values for the index of the activity concentration (I), radium equivalent activity (Raeq), and external hazard index (Hext) showed that these construction materials could be used without restrictions or concern about the equivalent dose limit (1 mSv/year). PMID:22280793

  16. 40 CFR 1065.205 - Performance specifications for measurement instruments.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... (CONTINUED) AIR POLLUTION CONTROLS ENGINE-TESTING PROCEDURES Measurement Instruments § 1065.205 Performance... or 0.2% of max K 0.2% of pt. K or 0.1% of max K 0.1% of max. Dewpoint sensor for intake air, PM.... Dilution air, inlet air, exhaust, and sample flow meters c n 1 s 1 Hz means of 5 Hz samples 2.5% of pt....

  17. Methods of and apparatus for radiation measurement, and specifically for in vivo radiation measurement

    DOEpatents

    Huffman, D.D.; Hughes, R.C.; Kelsey, C.A.; Lane, R.; Ricco, A.J.; Snelling, J.B.; Zipperian, T.E.

    1986-08-29

    Methods of and apparatus for in vivo radiation measurements rely on a MOSFET dosimeter of high radiation sensitivity which operates in both the passive mode to provide an integrated dose detector and active mode to provide an irradiation rate detector. A compensating circuit with a matched unirradiated MOSFET is provided to operate at a current designed to eliminate temperature dependence of the device. Preferably, the MOSFET is rigidly mounted in the end of a miniature catheter and the catheter is implanted in the patient proximate the radiation source.

  18. Atmospheric Radiation Measurement (ARM) Data from the ARM Specific Measurement Categories

    DOE Data Explorer

    The ARM Program gathers a wide variety of measurements from many different sources. Each day, the Data Archive stores and distributes large quantities of data collected from these sources. Scientists then use these data to research atmospheric radiation balance and cloud feedback processes, which are critical elements of global climate change. The huge archive of ARM data can be organized by measurement categories into six "collections:" Aerosols, Atmospheric Carbon, Atmospheric State, Cloud Properties, Radiometric, and Surface Properties. Clicking on one of the measurement categories leads to a page that breaks that category down into sub-categories. For example, "Aerosols" is broken down into Microphysical and Chemical Properties (with 9 subsets) and Optical and Radiative Properties (with 7 subsets). Each of the subset links, in turn, leads to detailed information pages and links to specific data streams. Users will be requested to create a password, but the data files are free for viewing and downloading. The ARM Archive physically resides at the Oak Ridge National Laboratory.

  19. Measuring segregation: an activity space approach

    PubMed Central

    Shaw, Shih-Lung

    2010-01-01

    While the literature clearly acknowledges that individuals may experience different levels of segregation across their various socio-geographical spaces, most measures of segregation are intended to be used in the residential space. Using spatially aggregated data to evaluate segregation in the residential space has been the norm and thus individual’s segregation experiences in other socio-geographical spaces are often de-emphasized or ignored. This paper attempts to provide a more comprehensive approach in evaluating segregation beyond the residential space. The entire activity spaces of individuals are taken into account with individuals serving as the building blocks of the analysis. The measurement principle is based upon the exposure dimension of segregation. The proposed measure reflects the exposure of individuals of a referenced group in a neighborhood to the populations of other groups that are found within the activity spaces of individuals in the referenced group. Using the travel diary data collected from the tri-county area in southeast Florida and the imputed racial–ethnic data, this paper demonstrates how the proposed segregation measurement approach goes beyond just measuring population distribution patterns in the residential space and can provide a more comprehensive evaluation of segregation by considering various socio-geographical spaces. PMID:21643546

  20. Measuring segregation: an activity space approach

    NASA Astrophysics Data System (ADS)

    Wong, David W. S.; Shaw, Shih-Lung

    2011-06-01

    While the literature clearly acknowledges that individuals may experience different levels of segregation across their various socio-geographical spaces, most measures of segregation are intended to be used in the residential space. Using spatially aggregated data to evaluate segregation in the residential space has been the norm and thus individual's segregation experiences in other socio-geographical spaces are often de-emphasized or ignored. This paper attempts to provide a more comprehensive approach in evaluating segregation beyond the residential space. The entire activity spaces of individuals are taken into account with individuals serving as the building blocks of the analysis. The measurement principle is based upon the exposure dimension of segregation. The proposed measure reflects the exposure of individuals of a referenced group in a neighborhood to the populations of other groups that are found within the activity spaces of individuals in the referenced group. Using the travel diary data collected from the tri-county area in southeast Florida and the imputed racial-ethnic data, this paper demonstrates how the proposed segregation measurement approach goes beyond just measuring population distribution patterns in the residential space and can provide a more comprehensive evaluation of segregation by considering various socio-geographical spaces.

  1. Recognition of military-specific physical activities with body-fixed sensors.

    PubMed

    Wyss, Thomas; Mäder, Urs

    2010-11-01

    The purpose of this study was to develop and validate an algorithm for recognizing military-specific, physically demanding activities using body-fixed sensors. To develop the algorithm, the first group of study participants (n = 15) wore body-fixed sensors capable of measuring acceleration, step frequency, and heart rate while completing six military-specific activities: walking, marching with backpack, lifting and lowering loads, lifting and carrying loads, digging, and running. The accuracy of the algorithm was tested in these isolated activities in a laboratory setting (n = 18) and in the context of daily military training routine (n = 24). The overall recognition rates during isolated activities and during daily military routine activities were 87.5% and 85.5%, respectively. We conclude that the algorithm adequately recognized six military-specific physical activities based on sensor data alone both in a laboratory setting and in the military training environment. By recognizing type of physical activities this objective method provides additional information on military-job descriptions. PMID:21121495

  2. General and Food-Specific Parenting: Measures and Interplay

    PubMed Central

    Sleddens, Ester; Gerards, Sanne; Gubbels, Jessica; Rodenburg, Gerda; Gevers, Dorus; van Assema, Patricia

    2013-01-01

    Abstract Background Parental influence on child food intake is typically conceptualized at three levels—parenting practices, feeding style, and parenting style. General parenting style is modeled at the most distal level of influence and food parenting practices are conceptualized as the most proximal level of influence. The goal of this article is to provide insights into contents and explanatory value of instruments that have been applied to assess food parenting practices, feeding style, and parenting style. Methods Measures of food parenting practices, feeding style, and parenting style were reviewed, compared, and contrasted with regard to contents, explanatory value, and interrelationships. Results Measures that are used in the field often fail to cover the full scope and complexity of food parenting. Healthy parenting dimensions have generally been found to be positively associated with child food intake (i.e., healthier dietary intake and less intake of energy-dense food products and sugar-sweetened beverages), but effect sizes are low. Evidence for the operation of higher-order moderation has been found, in which the impact of proximal parental influences is moderated by more distal levels of parenting. Conclusions Operationalizing parenting at different levels, while applying a contextual higher-order moderation approach, is advocated to have surplus value in understanding the complex process of parent–child interactions in the area of food intake. A research paradigm is presented that may guide future work regarding the conceptualization and modeling of parental influences on child dietary behavior. PMID:23944921

  3. Partial pressure measurements with an active spectrometer

    SciTech Connect

    Brooks, N.H.; Jensen, T.H.; Colchin, R.J.; Maingi, R.; Wade, M.R.; Finkenthal, D.F.; Naumenko, N.; Tugarinov, S.

    1998-07-01

    Partial pressure neutral ga measurements have been made using a commercial Penning gauge in conjunction with an active spectrometer. In prior work utilizing bandpass filters and conventional spectrometers, trace concentrations of the hydrogen isotopes H, D, T and of the noble gases He, Ne and Ar were determined from characteristic spectral lines in the light emitted by the neutral species of these elements. For all the elements mentioned, the sensitivity was limited by spectral contamination from a pervasive background of molecular hydrogen radiation. The active spectrometer overcomes this limitations by means of a digital lock-in method and correlation with reference spectra. Preliminary measurements of an admixture containing a trace amount of neon in deuterium show better than a factor of 20 improvement in sensitivity over conventional techniques. This can be further improved by correlating the relative intensities of multiple lines to sets of reference spectra.

  4. Direct measurement of specific membrane capacitance in neurons.

    PubMed

    Gentet, L J; Stuart, G J; Clements, J D

    2000-07-01

    The specific membrane capacitance (C(m)) of a neuron influences synaptic efficacy and determines the speed with which electrical signals propagate along dendrites and unmyelinated axons. The value of this important parameter remains controversial. In this study, C(m) was estimated for the somatic membrane of cortical pyramidal neurons, spinal cord neurons, and hippocampal neurons. A nucleated patch was pulled and a voltage-clamp step was applied. The exponential decay of the capacitative charging current was analyzed to give the total membrane capacitance, which was then divided by the observed surface area of the patch. C(m) was 0.9 microF/cm(2) for each class of neuron. To test the possibility that membrane proteins may alter C(m), embryonic kidney cells (HEK-293) were studied before and after transfection with a plasmid coding for glycine receptor/channels. The value of C(m) was indistinguishable in untransfected cells and in transfected cells expressing a high level of glycine channels, indicating that differences in transmembrane protein content do not significantly affect C(m). Thus, to a first approximation, C(m) may be treated as a "biological constant" across many classes of neuron. PMID:10866957

  5. Context-Specific Associations of Physical Activity and Sedentary Behavior With Cognition in Children.

    PubMed

    Aggio, Daniel; Smith, Lee; Fisher, Abigail; Hamer, Mark

    2016-06-15

    In the present study, we investigated how overall and specific domains of physical activity and sedentary behavior at the age of 7 years were associated with cognition at the age of 11 years in 8,462 children from the Millennium Cohort Study. Data were collected from 2001 to 2013. Participation in domains of physical activity and sedentary behavior at 7 years of age were reported. Activity levels were also measured objectively. Cognition was assessed using the British Ability Scales. General linear models were used to assess longitudinal associations of physical activity and sedentary behavior, measured both objectively and via self-report, with cognition. Analyses were adjusted for prespecified covariates. Sports/physical activity club attendance (B = 0.6, 95% confidence interval (CI): 0.2, 1.1), doing homework (B = 0.5, 95% CI: 0.0, 0.9), and objectively measured sedentary time (B = 0.8, 95% CI: 0.1, 1.4) at age 7 years were positively associated with cognition at age 11 years in final the models. Television viewing was negatively associated with cognition (B = -1.7, 95% CI: -2.4, -1.0), although the association was attenuated to the null after adjustments for baseline cognition. Objectively measured light physical activity was inversely associated with cognition (B = -0.7, 95% CI: -1.3, -0.1). Moderate-to-vigorous physical activity was also inversely associated with cognition in girls only (B = -1.1, 95% CI: -2.0, -0.3). Associations of physical activity and sedentary behavior with cognition appear to be context-specific in young people. PMID:27226249

  6. Context-Specific Associations of Physical Activity and Sedentary Behavior With Cognition in Children

    PubMed Central

    Aggio, Daniel; Smith, Lee; Fisher, Abigail; Hamer, Mark

    2016-01-01

    In the present study, we investigated how overall and specific domains of physical activity and sedentary behavior at the age of 7 years were associated with cognition at the age of 11 years in 8,462 children from the Millennium Cohort Study. Data were collected from 2001 to 2013. Participation in domains of physical activity and sedentary behavior at 7 years of age were reported. Activity levels were also measured objectively. Cognition was assessed using the British Ability Scales. General linear models were used to assess longitudinal associations of physical activity and sedentary behavior, measured both objectively and via self-report, with cognition. Analyses were adjusted for prespecified covariates. Sports/physical activity club attendance (B = 0.6, 95% confidence interval (CI): 0.2, 1.1), doing homework (B = 0.5, 95% CI: 0.0, 0.9), and objectively measured sedentary time (B = 0.8, 95% CI: 0.1, 1.4) at age 7 years were positively associated with cognition at age 11 years in final the models. Television viewing was negatively associated with cognition (B = −1.7, 95% CI: −2.4, −1.0), although the association was attenuated to the null after adjustments for baseline cognition. Objectively measured light physical activity was inversely associated with cognition (B = −0.7, 95% CI: −1.3, −0.1). Moderate-to-vigorous physical activity was also inversely associated with cognition in girls only (B = −1.1, 95% CI: −2.0, −0.3). Associations of physical activity and sedentary behavior with cognition appear to be context-specific in young people. PMID:27226249

  7. Sequence-specific DNA primer effects on telomerase polymerization activity.

    PubMed Central

    Lee, M S; Blackburn, E H

    1993-01-01

    The ribonucleoprotein enzyme telomerase synthesizes one strand of telomeric DNA by copying a template sequence within the RNA moiety of the enzyme. Kinetic studies of this polymerization reaction were used to analyze the mechanism and properties of the telomerase from Tetrahymena thermophila. This enzyme synthesizes TTGGGG repeats, the telomeric DNA sequence of this species, by elongating a DNA primer whose 3' end base pairs with the template-forming domain of the RNA. The enzyme was found to act nonprocessively with short (10- to 12-nucleotide) primers but to become processive as TTGGGG repeats were added. Variation of the 5' sequences of short primers with a common 3' end identified sequence-specific effects which are distinct from those involving base pairing of the 3' end of the primer with the RNA template and which can markedly induce enzyme activity by increasing the catalytic rate of the telomerase polymerization reaction. These results identify an additional mechanistic basis for telomere and DNA end recognition by telomerase in vivo. Images PMID:8413255

  8. Organ- and species-specific biological activity of rosmarinic acid.

    PubMed

    Iswandana, R; Pham, B T; van Haaften, W T; Luangmonkong, T; Oosterhuis, D; Mutsaers, H A M; Olinga, P

    2016-04-01

    Rosmarinic acid (RA), a compound found in several plant species, has beneficial properties, including anti-inflammatory and antibacterial effects. We investigated the toxicity, anti-inflammatory, and antifibrotic effects of RA using precision-cut liver slices (PCLS) and precision-cut intestinal slices (PCIS) prepared from human, mouse, and rat tissue. PCLS and PCIS were cultured up to 48h in the absence or presence of RA. Gene expression of the inflammatory markers: IL-6, IL-8/CXCL1/KC, and IL-1β, as well as the fibrosis markers: pro-collagen 1a1, heat shock protein 47, α-smooth muscle actin, fibronectin (Fn2) and plasminogen activator inhibitor-1 (PAI-1) were evaluated by qPCR. RA was only toxic in murine PCIS. RA failed to mitigate the inflammatory response in most models, while it clearly reduced IL-6 and CXCL1/KC gene expression in murine PCIS at non-toxic concentrations. With regard to fibrosis, RA decreased the gene levels of Fn2 and PAI-1 in murine PCLS, and Fn2 in murine PCIS. Yet, no effect was observed on the gene expression of fibrosis markers in human and rat PCIS. In conclusion, we observed clear organ- and species-specific effects of RA. RA had little influence on inflammation. However, our study further establishes RA as a potential candidate for the treatment of liver fibrosis. PMID:26804033

  9. Specific beta-adrenergic receptor binding of carazolol measured with PET

    SciTech Connect

    Berridge, M.S.; Nelson, A.D.; Zheng, L.

    1994-10-01

    Carazolol is a promising high-affinity beta-adrenergic receptor ligand for the noninvasive determination of beta receptor status using PET> Earlier investigations demonstrated specific receptor binding of carazolol in mice. These PET studies with S(-)-[2{double_prime}-{sup 11}C]carazolol in pigs were performed to explore the utility of the tracer for PET receptor studies. Tracer uptake in the heart and lung was measured by PET as a function of time. Receptors were blocked with propranolol and different doses of ICI 118,551 to estimate specific binding. Fluorine-18-1{double_prime}-Fluorocarazolol and the less active R-enantiomer of [{sup 11}C]-carazolol were also studied. Specific receptor binding was 75% of the total uptake in the heart, preventable and displaceable by propranolol. Dose-dependent competition showed that carazolol binds in vivo to {beta}{sub 1} and to {beta}{sub 2} subtypes. Uptake of the labeled R(=) enantiomer of carazolol was not receptor-specific. Carazolol labeled with {sup 11}C or {sup 18}F is a strong candidate for use in receptor estimation with PET. The in vivo observations were consistent with its known high affinity and slow receptor dissociation rate. Its high specific receptor uptake and low metabolism allow existing kinetic models to be applied for receptor measurements. The {sup 11}C label is convenient for repeated administrations, though {sup 13}F allowed the long observation periods necessary for measurement of the receptor dissociation rate. If needed, nonspecific uptake can be estimated without pharmacologic intervention by using the labeled R enantiomer. 32 refs., 11 figs.

  10. Neutron and proton activation measurements from Skylab

    NASA Technical Reports Server (NTRS)

    Fishman, G. J.

    1974-01-01

    Radioactivity induced by high-energy protons and secondary neutrons (from nuclear interactions) in various samples returned from different locations in Skylab was measured directly by gamma-ray spectroscopy measurements of decay gamma rays from the samples. Incident fluxes were derived from the activation measurements, using known nuclear cross-section. Neutron and proton flux values were found to range from 0.2 to 5 particles/sq cm-sec, depending on the energy range and location in Skylab. The thermal neutron flux was less than 0.07 neutrons/sq cm-sec. The results are useful for data analysis and planning of future high-energy astronomy experiments.

  11. Specific tension measurements in single soleus and medial gastrocnemius muscle fibers of the cat.

    PubMed

    Lucas, S M; Ruff, R L; Binder, M D

    1987-01-01

    Direct measurements of the sizes of and forces produced by single fibers of the cat soleus and medial gastrocnemius muscles were made to determine whether or not different fiber types have characteristically distinct specific tensions. Single fibers (5-mm lengths), whose sarcolemmas had been chemically removed using a 5-mM EGTA "skinning" solution, were attached to a photodiode force transducer. Each single fiber was first placed in "relaxing" solution (22 +/- 1 degrees C, pH 7.0, pCa 8), its sarcomere length set at 2.7 micron using its laser diffraction pattern, and its diameter measured using the calibrated graticule of a microscope eyepiece (+/- 2 micron). Subsequently, each fiber was transferred to an activating bathing solution (pCa 3.6) in which the fiber produced its maximum tension. The specific tension values for single soleus muscle fibers displayed a threefold range (1.19 to 3.53 kg/cm2) with a mean value of 2.30 +/- 0.61 (SD) kg/cm2 (N = 42). The medial gastrocnemius fibers studied had a fourfold range in specific tensions (1.05 to 4.47 kg/cm2) and a mean value of 2.42 +/- 0.61 (SD) kg/cm2 (N = 104). Many medial gastrocnemius fibers (N = 64) were type-identified using a standard actomyosin ATPase histochemical assay. Type I medial gastrocnemius fibers had mean specific tension values of 2.45 +/- 0.47 kg/cm2 (N = 18), whereas, type II single fibers had mean specific tension values of 2.43 +/- 0.67 kg/cm2 (N = 46). Our results suggest that there is no significant difference between the specific tensions of the different muscle fiber types within the cat medial gastrocnemius muscle. PMID:2947808

  12. Narration and Vividness as Measures of Event-Specificity in Autobiographical Memory

    ERIC Educational Resources Information Center

    Nelson, Kristin L.; Moskovitz, Damian J.; Steiner, Hans

    2008-01-01

    The event specificity of autobiographical memories refers to the degree to which retold memories include specific details about a unique personal experience from a variety of representational systems supported by different brain areas. This article proposes 2 text measures as indicators of event specificity: (a) a measure of temporal sequence in…

  13. Pyrimidine-specific 5' nucleotidase activity in bovine erythrocytes: effect of phlebotomy and lead poisoning

    SciTech Connect

    George, J.W.; Duncan, J.R.

    1982-01-01

    Erythrocyte pyrimidine-specific 5' nucleotidase (PY5'N) (E.C. 3.1.3.5) was measured in healthy, anemic, and lead-poisoned calves to determine whether low activity of PY5'N is associated with the propensity of cattle to develop basophilic stippling of erythrocytes. Low activity of PY5'N has been associated with basophilic stippling of erythrocytes in persons with inherited hemolytic anemia and with lead poisoning. A radiometric technique, using (/sup 14/C)cytidine monophosphate as the substrate, was used to measured PY5'N activity. The erythrocytes from 4 healthy calves had much lower activity (mean of 7.1 +/- 1.6 nmols of (/sup 14/C)cytidine monophosphate hydrolyzed/min/g of hemoglobin) than has been reported for human erythrocytes. The pH response curve of bovine PY5'N was similar to that of the human enzyme, with maximal activity around pH 7. Experimental hemorrhagic anemia in these calves increased PY5'N activity 6-to 7-fold, with peak activity occurring concomitantly with maximum reticulocytosis. Two of the calves were then given lead per os, and the PY5'N activity decreased within 24 hours to base-line values. In the 2 other calves not given lead, the PY5'N activity declined slowly, but did not reach base-line values after 14 days.

  14. Patient specific proteolytic activity of monocyte-derived macrophages and osteoclasts predicted with temporal kinase activation states during differentiation

    PubMed Central

    Park, Keon-Young; Li, Weiwei A.; Platt, Manu O.

    2012-01-01

    Patient-to-patient variability in disease progression continues to complicate clinical decisions of treatment regimens for cardiovascular diseases, metastatic cancers and osteoporosis. Here, we investigated if monocytes, circulating white blood cells that enter tissues and contribute to disease progression by differentiating into macrophages or osteoclasts, could be useful in understanding this variability. Monocyte-derived macrophages and osteoclasts produce cysteine cathepsins, powerful extracellular matrix proteases which have been mechanistically linked to accelerated atherosclerotic, osteoporotic, and tumor progression. We hypothesized that multivariate analysis of temporal kinase activation states during monocyte differentiation could predict cathepsin proteolytic responses of monocyte-derived macrophages and osteoclasts in a patient-specific manner. Freshly isolated primary monocytes were differentiated with M-CSF or RANKL into macrophages or osteoclasts, respectively, and phosphorylation of ERK1/2, Akt, p38 MAPK, JNK, c-jun, and IκB-α were measured at days 1, 3, 6, and 9. In parallel, cell diameters and numbers of nuclei were measured, and multiplex cathepsin zymography was used to quantify cathepsins K, L, S, and V activity from cell extracts and conditioned media. There was extensive patient-to-patient variability in temporal kinase activation states, cell morphologies, and cathepsin K, L, S, and V proteolytic activity. Partial least squares regression models trained with temporal kinase activation states successfully predicted patient-specific morphological characteristics (mean cell diameter and number of nuclei) and patient-specific cathepsin proteolytic activity with predictability as high as 95%, even with the challenge of incorporating the complex, unknown cues from individual patients’ unique genetic and biochemical backgrounds. This personalized medicine approach considers patient variability in kinase signals to predict cathepsin activity

  15. Roles of s3 site residues of nattokinase on its activity and substrate specificity.

    PubMed

    Wu, Shuming; Feng, Chi; Zhong, Jin; Huan, Liandong

    2007-09-01

    Nattokinase (Subtilisin NAT, NK) is a bacterial serine protease with high fibrinolytic activity. To probe their roles on protease activity and substrate specificity, three residues of S3 site (Gly(100), Ser(101) and Leu(126)) were mutated by site-directed mutagenesis. Kinetics parameters of 20 mutants were measured using tetrapeptides as substrates, and their fibrinolytic activities were determined by fibrin plate method. Results of mutation analysis showed that Gly(100) and Ser(101) had reverse steric and electrostatic effects. Residues with bulky or positively charged side chains at position 100 decreased the substrate binding and catalytic activity drastically, while residues with the same characters at position 101 could obviously enhance protease and fibrinolytic activity of NK. Mutation of Leu(126) might impair the structure of the active cleft and drastically decreased the activity of NK. Kinetics studies of the mutants showed that S3 residues were crucial to keep protease activity while they moderately affected substrate specificity of NK. The present study provided some original insight into the P3-S3 interaction in NK and other subtilisins, as well as showed successful protein engineering cases to improve NK as a potential therapeutic agent. PMID:17673485

  16. Utilizing Electroencephalography Measurements for Comparison of Task-Specific Neural Efficiencies: Spatial Intelligence Tasks.

    PubMed

    Call, Benjamin J; Goodridge, Wade; Villanueva, Idalis; Wan, Nicholas; Jordan, Kerry

    2016-01-01

    Spatial intelligence is often linked to success in engineering education and engineering professions. The use of electroencephalography enables comparative calculation of individuals' neural efficiency as they perform successive tasks requiring spatial ability to derive solutions. Neural efficiency here is defined as having less beta activation, and therefore expending fewer neural resources, to perform a task in comparison to other groups or other tasks. For inter-task comparisons of tasks with similar durations, these measurements may enable a comparison of task type difficulty. For intra-participant and inter-participant comparisons, these measurements provide potential insight into the participant's level of spatial ability and different engineering problem solving tasks. Performance on the selected tasks can be analyzed and correlated with beta activities. This work presents a detailed research protocol studying the neural efficiency of students engaged in the solving of typical spatial ability and Statics problems. Students completed problems specific to the Mental Cutting Test (MCT), Purdue Spatial Visualization test of Rotations (PSVT:R), and Statics. While engaged in solving these problems, participants' brain waves were measured with EEG allowing data to be collected regarding alpha and beta brain wave activation and use. The work looks to correlate functional performance on pure spatial tasks with spatially intensive engineering tasks to identify the pathways to successful performance in engineering and the resulting improvements in engineering education that may follow. PMID:27584838

  17. Noninvasive ambulatory measurement system of cardiac activity.

    PubMed

    Pino, Esteban J; Chavez, Javier A P; Aqueveque, Pablo

    2015-08-01

    This work implements a noninvasive system that measures the movements caused by cardiac activity. It uses unobtrusive Electro-Mechanical Films (EMFi) on the seat and on the backrest of a regular chair. The system detects ballistocardiogram (BCG) and respiration movements. Real data was obtained from 54 volunteers. 19 of them were measured in the laboratory and 35 in a hospital waiting room. Using a BIOPAC acquisition system, the ECG was measured simultaneously to the BCG for comparison. Wavelet Transform (WT) is a better option than Empirical Mode Decomposition (EMD) for signal extraction and produces higher effective measurement time. In the laboratory, the best results are obtained on the seat. The correlation index was 0.9800 and the Bland-Altman limits of agreement were 0.7136 ± 4.3673 [BPM]. In the hospital waiting room, the best results are also from the seat sensor. The correlation index was 0.9840, and the limits of agreement were 0.4386 ± 3.5884 [BPM]. The system is able to measure BCG in an unobtrusive way and determine the cardiac frequency with high precision. It is simple to use, which means the system can easily be used in non-standard settings: resting in a chair or couch, at the gym, schools or in a hospital waiting room, as shown. PMID:26738057

  18. Tissue-specific inhibition and recovery of esterase activities in Lumbricus terrestris experimentally exposed to chlorpyrifos.

    PubMed

    Vejares, Sandra González; Sabat, Pablo; Sanchez-Hernandez, Juan C

    2010-04-01

    Exposure and effect assessment of organophosphate (OP) pesticides generally involves the use of cholinesterase (ChE) inhibition. In earthworm, this enzyme activity is often measured in homogenates from the whole organism. Here we examine the tissue-specific response of ChE and carboxylesterase (CE) activities in Lumbricus terrestris experimentally exposed to chlorpyrifos-spiked field soils. Esterases were measured in different gut segments and in the seminal vesicles of earthworms following acute exposure (2 d) to the OP and during 35d of a recovery period. We found that inhibition of both esterase activities was dependent on the tissue. Cholinesterase activity decreased in the pharynx, crop, foregut and seminal vesicles in a concentration-dependent way, whereas CE activity (4-nitrophenyl valerate) was strongly inhibited in these tissues. Gizzard CE activity was not inhibited by the OP, even an increase of enzyme activity was evident during the recovery period. These results suggest that both esterases should be determined jointly in selected tissues of earthworms. Moreover, the high levels of gut CE activity and its inhibition and recovery dynamic following OP exposure suggest that this esterase could play an important role as an enzymatic barrier against OP uptake from the ingested contaminated soil. PMID:20045489

  19. Comparisons of activity measurements with radionuclide calibrators.

    PubMed

    Oropesa, P; Hernández, A T; Serra, R; Martínez, E; Varela, C

    2003-01-01

    The correct administration to a patient of the prescribed activity of a radiopharmaceutical is an important factor to ensure the confidence in the diagnosis or the therapeutic efficiency, while at the same time keeping the unnecessary human exposure as low as possible. Comparisons of activity measurements for 131I, 201Tl and 99mTc with radionuclide calibrators were organized the first time in Cuba during 2002 with the aim of obtaining information about the quality of administration of radiopharmaceuticals. Ten Cuban nuclear medicine departments and the laboratories involved in the production of these kinds of compounds participated in the comparison runs. The results presented in this paper facilitated the identification of several problems and initiated corrective actions. In addition, they indicate the necessity of establishing Quality Systems in nuclear medicine in Cuba. PMID:14622940

  20. Photocatalytic Active Radiation Measurements and Use

    NASA Technical Reports Server (NTRS)

    Davis, Bruce A.; Underwood, Lauren W.

    2011-01-01

    Photocatalytic materials are being used to purify air, to kill microbes, and to keep surfaces clean. A wide variety of materials are being developed, many of which have different abilities to absorb various wavelengths of light. Material variability, combined with both spectral illumination intensity and spectral distribution variability, will produce a wide range of performance results. The proposed technology estimates photocatalytic active radiation (PcAR), a unit of radiation that normalizes the amount of light based on its spectral distribution and on the ability of the material to absorb that radiation. Photocatalytic reactions depend upon the number of electron-hole pairs generated at the photocatalytic surface. The number of electron-hole pairs produced depends on the number of photons per unit area per second striking the surface that can be absorbed and whose energy exceeds the bandgap of the photocatalytic material. A convenient parameter to describe the number of useful photons is the number of moles of photons striking the surface per unit area per second. The unit of micro-einsteins (or micromoles) of photons per m2 per sec is commonly used for photochemical and photoelectric-like phenomena. This type of parameter is used in photochemistry, such as in the conversion of light energy for photosynthesis. Photosynthetic response correlates with the number of photons rather than by energy because, in this photochemical process, each molecule is activated by the absorption of one photon. In photosynthesis, the number of photons absorbed in the 400 700 nm spectral range is estimated and is referred to as photosynthetic active radiation (PAR). PAR is defined in terms of the photosynthetic photon flux density measured in micro-einsteins of photons per m2 per sec. PcAR is an equivalent, similarly modeled parameter that has been defined for the photocatalytic processes. Two methods to measure the PcAR level are being proposed. In the first method, a calibrated

  1. Regulation of platelet activating factor receptor coupled phosphoinositide-specific phospholipase C activity

    SciTech Connect

    Morrison, W.J.

    1988-01-01

    The major objectives of this study were two-fold. The first was to establish whether binding of platelet activating factor (PAF) to its receptor was integral to the stimulation of polyphosphoinositide-specific phospholipase C (PLC) in rabbit platelets. The second was to determine regulatory features of this receptor-coupled mechanism. ({sup 3}H)PAF binding demonstrated two binding sites, a high affinity site with a inhibitory constant (Ki) of 2.65 nM and a low affinity site with a Ki of 0.80 {mu}M. PAF receptor coupled activation of phosphoinositide-specific PLC was studied in platelets which were made refractory, by short term pretreatments, to either PAF or thrombin. Saponin-permeabilized rabbit platelets continue to regulate the mechanism(s) coupling PAF receptors to PLC stimulation. However, TRP{gamma}S and GDP{beta}S, which affect guanine nucleotide regulatory protein functions, were unable to modulate the PLC activity to any appreciable extent as compared to PAF. The possible involvement of protein kinase C (PKC) activation in regulating PAF-stimulated PLC activity was studied in rabbit platelets pretreated with staurosporine followed by pretreatments with PAF or phorbol 12-myristate 13-acetate (PMA).

  2. Non-specific activation of the epithelial sodium channel by the CFTR chloride channel

    PubMed Central

    Nagel, Georg; Szellas, Tanjef; Riordan, John R.; Friedrich, Thomas; Hartung, Klaus

    2001-01-01

    The genetic disease cystic fibrosis is caused by mutation of the gene coding for the cystic fibrosis transmembrane conductance regulator (CFTR). Controversial studies reported regulation of the epithelial sodium channel (ENaC) by CFTR. We found that uptake of 22Na+ through ENaC is modulated by activation of CFTR in oocytes, coexpressing CFTR and ENaC, depending on extracellular chloride concentration. Furthermore we found that the effect of CFTR activation could be mimicked by other chloride channels. Voltage– and patch–clamp measurements, however, showed neither stimulation nor inhibition of ENaC-mediated conductance by activated CFTR. We conclude that the observed modulation of 22Na+ uptake by activated CFTR is due to the effect of CFTR-mediated chloride conductance on the membrane potential. These findings argue against the notion of a specific influence of CFTR on ENaC and emphasize the chloride channel function of CFTR. PMID:11266369

  3. Non-specific activation of the epithelial sodium channel by the CFTR chloride channel.

    PubMed

    Nagel, G; Szellas, T; Riordan, J R; Friedrich, T; Hartung, K

    2001-03-01

    The genetic disease cystic fibrosis is caused by mutation of the gene coding for the cystic fibrosis transmembrane conductance regulator (CFTR). Controversial studies reported regulation of the epithelial sodium channel (ENaC) by CFTR. We found that uptake of (22)Na(+) through ENaC is modulated by activation of CFTR in oocytes, coexpressing CFTR and ENaC, depending on extracellular chloride concentration. Furthermore we found that the effect of CFTR activation could be mimicked by other chloride channels. Voltage- and patch-clamp measurements, however, showed neither stimulation nor inhibition of ENaC-mediated conductance by activated CFTR. We conclude that the observed modulation of (22)Na(+) uptake by activated CFTR is due to the effect of CFTR-mediated chloride conductance on the membrane potential. These findings argue against the notion of a specific influence of CFTR on ENaC and emphasize the chloride channel function of CFTR. PMID:11266369

  4. Electrochemical assay of active prostate-specific antigen (PSA) using ferrocene-functionalized peptide probes

    SciTech Connect

    Zhao, Ning; He, Yuqing; Mao, Xun; Sun, Yuhan; Zhang, Xibao; Li, Chen-Zhong; Lin, Yuehe; Liu, Guodong

    2010-03-24

    This paper presents a novel approach to electrochemically determine enzymatically active PSA using ferrocene-functionalized helix peptide (CHSSLKQK). The principle of electrochemical measurement is based on the specific proteolytic cleavage events of the FC-peptide on the gold electrode surface in the presence of PSA, resulting the change of the current signal of the electrode. The percentage of the decreased current is linear with the concentration of active PSA at the range of 0.5-40 ng/mL with a detection limit of 0.2 ng/mL. The direct transduction of peptide cleavage events into an electrical signal provides a simple, sensitive method for detecting the enzymatic activity of PSA and determining the active PSA concentration.

  5. Synthesis of a high specific activity methyl sulfone tritium isotopologue of fevipiprant (NVP-QAW039).

    PubMed

    Luu, Van T; Goujon, Jean-Yves; Meisterhans, Christian; Frommherz, Matthias; Bauer, Carsten

    2015-05-15

    The synthesis of a triple tritiated isotopologue of the CRTh2 antagonist NVP-QAW039 (fevipiprant) with a specific activity >3 TBq/mmol is described. Key to the high specific activity is the methylation of a bench-stable dimeric disulfide precursor that is in situ reduced to the corresponding thiol monomer and methylated with [(3)H3]MeONos having per se a high specific activity. The high specific activity of the tritiated active pharmaceutical ingredient obtained by a build-up approach is discussed in the light of the specific activity usually to be expected if hydrogen tritium exchange methods were applied. PMID:25881897

  6. Measuring Homework Completion in Behavioral Activation

    PubMed Central

    Busch, Andrew M.; Uebelacker, Lisa A.; Kalibatseva, Zornitsa; Miller, Ivan W.

    2010-01-01

    The aim of this study was to develop and validate an observer-based coding system for the characterization and completion of homework assignments during Behavioral Activation (BA). Existing measures of homework completion are generally unsophisticated, and there is no current measure of homework completion designed to capture the particularities of BA. The tested scale sought to capture the type of assignment, realm of functioning targeted, extent of completion, and assignment difficulty. Homework assignments were drawn from 12 (mean age = 48, 83% female) clients in two trials of a 10-session BA manual targeting treatment-resistant depression in primary care. The two coders demonstrated acceptable or better reliability on most codes, and unreliable codes were dropped from the proposed scale. In addition, correlations between homework completion and outcome were strong, providing some support for construct validity. Ultimately, this line of research aims to develop a user-friendly, reliable measure of BA homework completion that can be completed by a therapist during session. PMID:20562324

  7. 50 CFR 654.27 - Specifically authorized activities.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE STONE CRAB FISHERY OF THE GULF OF MEXICO Management... acquisition of information and data, activities otherwise prohibited by the regulations in this part....

  8. 50 CFR 654.27 - Specifically authorized activities.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE STONE CRAB FISHERY OF THE GULF OF MEXICO Management... acquisition of information and data, activities otherwise prohibited by the regulations in this part....

  9. Toward a Cultural Competence Measure for Social Work with Specific Populations.

    ERIC Educational Resources Information Center

    Boyle, David P.; Springer, Alyson

    2001-01-01

    Four measures of cultural competency in the social work profession are analyzed and found to measure global constructs that may not be applicable to specific cultures. The development of culture-specific instruments is proposed, from which a general cultural competence instrument can be distilled. (Contains 32 references.) (TD)

  10. Comparing Beginning Teachers' Instructional Quality Growth on Subject-Specific and Global Measures

    ERIC Educational Resources Information Center

    Neergaard, Laura; Smith, Tom

    2012-01-01

    Observation measures of instructional quality tend to fall into two broad categories--those for use across subject areas and those intended for use in specific subject areas. The move toward content-specific measures is a result of research suggesting that effective teaching looks different across subject areas and that both content knowledge and…

  11. Activity of increased specific and non-specific esterases and glutathione transferases associated with resistance to permethrin in pediculus humanus capitis (phthiraptera: pediculidae) from Argentina.

    PubMed

    Barrios, Silvia; Zerba, Eduardo; Picollo, Maria I; Audino, Paola Gonzalez

    2010-01-01

    Enhanced metabolism by oxidative enzymes is a major cause of pyrethroid resistance in insects. In this work, we evaluated the role of specific and non-specific esterases in head louse populations from Buenos Aires with different levels of resistance to permethrin. As esterase activity is substrate-dependent, four different esters were used as unspecific substrates in order to obtain a better characterization of the possible role of these enzymes in the resistance phenomenon. The unspecific substrates were phenylthioacetate, 1- and 2-naphtyl-acetate, and p-nitrophenyl acetate. A 7-coumaryl permethrate was synthesized and used as a specific substrate to measure pyrethroid esterases by a very sensitive microfluorometric method. The results on pyrethroid esterase activity obtained with this substrate showed that these enzymes contribute to the detoxifying activity in resistant populations, although no correlation was found between pyrethroid esterase activity and resistance ratios. In this study, we established that the activity of esterase against specific and non-specific substrates is increased in pyrethroid-resistant populations of head lice from Buenos Aires. Also, dichlorodiphenyltrichloroethane (DDT) resistance values demonstrated that there is a DDT cross-resistance phenomenon in pyrethroid-resistant head louse populations and suggested that an alteration in the receptor of the nervous system (kdr gen) is a key factor of the resistance phenomena in these head louse populations. PMID:19921258

  12. Specification of High Activity Gamma-Ray Sources.

    ERIC Educational Resources Information Center

    International Commission on Radiation Units and Measurements, Washington, DC.

    The report is concerned with making recommendations for the specifications of gamma ray sources, which relate to the quantity of radioactive material and the radiation emitted. Primary consideration is given to sources in teletherapy and to a lesser extent those used in industrial radiography and in irradiation units used in industry and research.…

  13. 10 CFR 810.8 - Activities requiring specific authorization.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... technology for an activity in any foreign country. (c) Engaging in or providing assistance or training in any... plutonium, or the production of heavy water; (2) Constructing, fabricating, operating, or maintaining...

  14. 10 CFR 810.8 - Activities requiring specific authorization.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... technology for an activity in any foreign country. (c) Engaging in or providing assistance or training in any... plutonium, or the production of heavy water; (2) Constructing, fabricating, operating, or maintaining...

  15. Measuring Cysteine Cathepsin Activity to Detect Lysosomal Membrane Permeabilization.

    PubMed

    Repnik, Urška; Česen, Maruša Hafner; Turk, Boris

    2016-01-01

    During lysosomal membrane permeabilization (LMP), lysosomal lumenal contents can be released into the cytosol. Small molecules are more likely to be released, and cysteine cathepsins, with mature forms possessing a mass of 25-30 kDa, are among the smallest lumenal lysosomal enzymes. In addition, specific substrates for cysteine cathepsins are available to investigators, and therefore the measurement of the cathepsin activity as a hallmark of LMP works well. Here, we present a protocol for measuring the activity of these enzymes after selective plasma membrane permeabilization with a low concentration of digitonin and after total cell membrane lysis with a high concentration of digitonin. A fluorogenic substrate can be added either directly to the well with lysed cells to show LMP or to the cell-free extract to show that the lysosomal membrane has been sufficiently destabilized to allow the translocation of lysosomal enzymes. Although the content of lysosomal cysteine cathepsins differs between cell lines, this method has general applicability, is sensitive, and has high throughput. The presented protocol shows how to measure cysteine cathepsin activity in the presence of lysed cells and also in cell-free extracts. Depending on the aim of the study, one or both types of measurements can be performed. PMID:27140915

  16. Assessing specific deterrence effects of increased speeding penalties using four measures of recidivism.

    PubMed

    Watson, B; Siskind, V; Fleiter, J J; Watson, A; Soole, D

    2015-11-01

    Traffic law enforcement sanctions can impact on road user behaviour through general and specific deterrence mechanisms. The manner in which specific deterrence can influence recidivist behaviour can be conceptualised in different ways. While any reduction in speeding will have road safety benefits, the ways in which a 'reduction' is determined deserves greater methodological attention and has implications for countermeasure evaluation more generally. The primary aim of this research was to assess the specific deterrent impact of penalty increases for speeding offences in Queensland, Australia, in 2003 on two cohorts of drivers detected for speeding prior to and after the penalty changes were investigated. Since the literature is relatively silent on how to assess recidivism in the speeding context, the secondary research aim was to contribute to the literature regarding ways to conceptualise and measure specific deterrence in the speeding context. We propose a novel way of operationalising four measures which reflect different ways in which a specific deterrence effect could be conceptualised: (1) the proportion of offenders who re-offended in the follow up period; (2) the overall frequency of re-offending in the follow up period; (3) the length of delay to re-offence among those who re-offended; and (4) the average number of re-offences during the follow up period among those who re-offended. Consistent with expectations, results suggested an absolute deterrent effect of penalty changes, as evidenced by significant reductions in the proportion of drivers who re-offended and the overall frequency of re-offending, although effect sizes were small. Contrary to expectations, however, there was no evidence of a marginal specific deterrent effect among those who re-offended, with a significant reduction in the length of time to re-offence and no significant change in the average number of offences committed. Additional exploratory analyses investigating potential

  17. Protein Translation Activity: A New Measure of Host Immune Cell Activation.

    PubMed

    Seedhom, Mina O; Hickman, Heather D; Wei, Jiajie; David, Alexandre; Yewdell, Jonathan W

    2016-08-15

    We describe the in vivo ribopuromycylation (RPM) method, which uses a puromycin-specific Ab to fluorescently label ribosome-bound puromycylated nascent chains, enabling measurement of translational activity via immunohistochemistry or flow cytometry. Tissue staining provides a unique view of virus-induced activation of adaptive, innate, and stromal immune cells. RPM flow precisely quantitates virus-induced activation of lymphocytes and innate immune cells, and it provides a unique measure of immune cell deactivation and quiescence. Using RPM we find that high endothelial cells in draining lymph nodes rapidly increase translation in the first day of vaccinia virus infection. We also find a population of constitutively activated splenic T cells in naive mice and further that most bone marrow T cells activate 3 d after vaccinia virus infection. Bone marrow T cell activation is nonspecific, IL-12-dependent, and induces innate memory T cell phenotypic markers. Thus, RPM measures translational activity to uniquely identify cell populations that participate in the immune response to pathogens, other foreign substances, and autoantigens. PMID:27385780

  18. How to produce high specific activity tin-117m using alpha particle beam.

    PubMed

    Duchemin, C; Essayan, M; Guertin, A; Haddad, F; Michel, N; Métivier, V

    2016-09-01

    Tin-117m is an interesting radionuclide for both diagnosis and therapy, thanks to the gamma-ray and electron emissions, respectively, resulting from its decay to tin-117g. The high specific activity of tin-117m is required in many medical applications, and it can be obtained using a high energy alpha particle beam and a cadmium target. The experiments performed at the ARRONAX cyclotron (Nantes, France) using an alpha particle beam delivered at 67.4MeV provide a measurement of the excitation function of the Cd-nat(α,x)Sn-117m reaction and the produced contaminants. The Cd-116(α,3n)Sn-117m production cross section has been deduced from these experimental results using natural cadmium. Both production yield and specific activity as a function of the projectile energy have been calculated. These informations help to optimize the irradiation conditions to produce tin-117m with the required specific activity using α particles with a cadmium target. PMID:27344526

  19. A dynamic model for generating actuator specifications for small arms barrel active stabilization

    NASA Astrophysics Data System (ADS)

    Pathak, Anupam; Brei, Diann; Luntz, Jonathan; Lavigna, Chris

    2006-03-01

    Due to stresses encountered in combat, it is known that soldier marksmanship noticeably decreases regardless of prior training. Active stabilization systems in small arms have potential to address this problem to increase soldier survivability and mission effectiveness. The key to success is proper actuator design, but this is highly dependent on proper specification which is challenging due to the human/weapon interaction. This paper presents a generic analytical dynamic model which is capable of defining the necessary actuation specifications for a wide range of small arms platforms. The model is unique because it captures the human interface--shoulder and arm--that introduces the jitter disturbance in addition to the geometry, inertial properties and active stabilization stiffness of the small arms platform. Because no data to date is available for actual shooter-induced disturbance in field conditions, a method is given using the model to back-solve from measured shooting range variability data the disturbance amplitude information relative to the input source (arm or shoulder). As examples of the applicability of the model to various small arms systems, two different weapon systems were investigated: the M24 sniper weapon and the M16 assault rifle. In both cases, model based simulations provided valuable insight into impact on the actuation specifications (force, displacement, phase, frequency) due to the interplay of the human-weapon-active stabilization interface including the effect of shooter-disturbance frequency, disturbance location (shoulder vs. arm), and system parameters (stiffness, barrel rotation).

  20. 21 CFR 331.11 - Listing of specific active ingredients.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... ion; maximum daily dosage limit 200 mEq. for persons up to 60 years old and 100 mEq. for persons 60...., 8 grams calcium carbonate). (e) Citrate-containing active ingredients: Citrate ion, as citric acid... effervescent preparation); maximum daily dosage limit 200 mEq. of bicarbonate ion for persons up to 60...

  1. 21 CFR 331.11 - Listing of specific active ingredients.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... ion; maximum daily dosage limit 200 mEq. for persons up to 60 years old and 100 mEq. for persons 60...., 8 grams calcium carbonate). (e) Citrate-containing active ingredients: Citrate ion, as citric acid... effervescent preparation); maximum daily dosage limit 200 mEq. of bicarbonate ion for persons up to 60...

  2. 21 CFR 331.11 - Listing of specific active ingredients.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... ion; maximum daily dosage limit 200 mEq. for persons up to 60 years old and 100 mEq. for persons 60...., 8 grams calcium carbonate). (e) Citrate-containing active ingredients: Citrate ion, as citric acid... effervescent preparation); maximum daily dosage limit 200 mEq. of bicarbonate ion for persons up to 60...

  3. 21 CFR 331.11 - Listing of specific active ingredients.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... ion; maximum daily dosage limit 200 mEq. for persons up to 60 years old and 100 mEq. for persons 60...., 8 grams calcium carbonate). (e) Citrate-containing active ingredients: Citrate ion, as citric acid... effervescent preparation); maximum daily dosage limit 200 mEq. of bicarbonate ion for persons up to 60...

  4. 21 CFR 331.11 - Listing of specific active ingredients.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... ion; maximum daily dosage limit 200 mEq. for persons up to 60 years old and 100 mEq. for persons 60...., 8 grams calcium carbonate). (e) Citrate-containing active ingredients: Citrate ion, as citric acid... effervescent preparation); maximum daily dosage limit 200 mEq. of bicarbonate ion for persons up to 60...

  5. Nanoliposomes containing Eucalyptus citriodora as antibiotic with specific antimicrobial activity.

    PubMed

    Lin, Lin; Cui, Haiying; Zhou, Hui; Zhang, Xuejing; Bortolini, Christian; Chen, Menglin; Liu, Lei; Dong, Mingdong

    2015-02-14

    Bacterial infections are a serious issue for public health and represent one of the major challenges of modern medicine. In this work, a selective antimicrobial strategy based on triggering of pore-forming toxin, which is secreted by infective bacteria, was designed to fight Staphylococcus aureus. The antimicrobial activity is realized by employing Eucalyptus citriodora oil as antibiotic which in this study is encapsulated in nanoliposomes. PMID:25573466

  6. Electromagnetic Measurements in an Active Oilfield Environment

    NASA Astrophysics Data System (ADS)

    Schramm, K. A.; Aldridge, D. F.; Bartel, L. C.; Knox, H. A.; Weiss, C. J.

    2015-12-01

    An important issue in oilfield development pertains to mapping and monitoring of the fracture distributions (either natural or man-made) controlling subsurface fluid flow. Although microseismic monitoring and analysis have been used for this purpose for several decades, there remain several ambiguities and uncertainties with this approach. We are investigating a novel electromagnetic (EM) technique for detecting and mapping hydraulic fractures in a petroleum reservoir by injecting an electrically conductive contrast agent into an open fracture. The fracture is subsequently illuminated by a strong EM field radiated by a large engineered antenna. Specifically, a grounded electric current source is applied directly to the steel casing of the borehole, either at/near the wellhead or at a deep downhole point. Transient multicomponent EM signals (both electric and magnetic) scattered by the conductivity contrast are then recorded by a surface receiver array. We are presently utilizing advanced 3D numerical modeling algorithms to accurately simulate fracture responses, both before and after insertion of the conductive contrast agent. Model results compare favorably with EM field data recently acquired in a Permian Basin oilfield. However, extraction of the very-low-amplitude fracture signatures from noisy data requires effective noise suppression strategies such as long stacking times, rejection of outliers, and careful treatment of natural magnetotelluric fields. Dealing with the ever-present "episodic EM noise" typical in an active oilfield environment (associated with drilling, pumping, machinery, traffic, etc.) constitutes an ongoing problem. Sandia National Laboratories is a multi-program laboratory managed and operated by Sandia Corporation, a wholly owned subsidiary of Lockheed Martin Corporation, for the US Department of Energy's National Nuclear Security Administration under contract DE-AC04-94AL85000.

  7. Repetitive Transcranial Magnetic Stimulation Activates Specific Regions in Rat Brain

    NASA Astrophysics Data System (ADS)

    Ji, Ru-Rong; Schlaepfer, Thomas E.; Aizenman, Carlos D.; Epstein, Charles M.; Qiu, Dike; Huang, Justin C.; Rupp, Fabio

    1998-12-01

    Repetitive transcranial magnetic stimulation (rTMS) is a noninvasive technique to induce electric currents in the brain. Although rTMS is being evaluated as a possible alternative to electroconvulsive therapy for the treatment of refractory depression, little is known about the pattern of activation induced in the brain by rTMS. We have compared immediate early gene expression in rat brain after rTMS and electroconvulsive stimulation, a well-established animal model for electroconvulsive therapy. Our result shows that rTMS applied in conditions effective in animal models of depression induces different patterns of immediate-early gene expression than does electroconvulsive stimulation. In particular, rTMS evokes strong neural responses in the paraventricular nucleus of the thalamus (PVT) and in other regions involved in the regulation of circadian rhythms. The response in PVT is independent of the orientation of the stimulation probe relative to the head. Part of this response is likely because of direct activation, as repetitive magnetic stimulation also activates PVT neurons in brain slices.

  8. Small business activity does not measure entrepreneurship.

    PubMed

    Henrekson, Magnus; Sanandaji, Tino

    2014-02-01

    Entrepreneurship policy mainly aims to promote innovative Schumpeterian entrepreneurship. However, the rate of entrepreneurship is commonly proxied using quantity-based metrics, such as small business activity, the self-employment rate, or the number of startups. We argue that those metrics give rise to misleading inferences regarding high-impact Schumpeterian entrepreneurship. To unambiguously identify high-impact entrepreneurs we focus on self-made billionaires (in US dollars) who appear on Forbes Magazine's list and who became wealthy by founding new firms. We identify 996 such billionaire entrepreneurs in 50 countries in 1996-2010, a systematic cross-country study of billionaire entrepreneurs. The rate of billionaire entrepreneurs correlates negatively with self-employment, small business ownership, and firm startup rates. Countries with higher income, higher trust, lower taxes, more venture capital investment, and lower regulatory burdens have higher billionaire entrepreneurship rates but less self-employment. Despite its limitations, the number of billionaire entrepreneurs appears to be a plausible cross-country measure of Schumpeterian entrepreneurship. PMID:24449873

  9. Small business activity does not measure entrepreneurship

    PubMed Central

    Henrekson, Magnus; Sanandaji, Tino

    2014-01-01

    Entrepreneurship policy mainly aims to promote innovative Schumpeterian entrepreneurship. However, the rate of entrepreneurship is commonly proxied using quantity-based metrics, such as small business activity, the self-employment rate, or the number of startups. We argue that those metrics give rise to misleading inferences regarding high-impact Schumpeterian entrepreneurship. To unambiguously identify high-impact entrepreneurs we focus on self-made billionaires (in US dollars) who appear on Forbes Magazine’s list and who became wealthy by founding new firms. We identify 996 such billionaire entrepreneurs in 50 countries in 1996–2010, a systematic cross-country study of billionaire entrepreneurs. The rate of billionaire entrepreneurs correlates negatively with self-employment, small business ownership, and firm startup rates. Countries with higher income, higher trust, lower taxes, more venture capital investment, and lower regulatory burdens have higher billionaire entrepreneurship rates but less self-employment. Despite its limitations, the number of billionaire entrepreneurs appears to be a plausible cross-country measure of Schumpeterian entrepreneurship. PMID:24449873

  10. Physiotherapy treatment for atraumatic recurrent shoulder instability: early results of a specific exercise protocol using pathology-specific outcome measures

    PubMed Central

    Smith, Benjamin E; Osborne, Sally E; Wilkes, Sally R

    2015-01-01

    Background Recurrent shoulder instability is usually caused by a traumatic event resulting in structural pathology, although a small subgroup of patients experience symptomatic recurrent shoulder instability without trauma. These patients are usually treated non-operatively but limited evidence exists regarding effective conservative management. In particular, there is a lack of reproducible exercise regimes and none that have been tested with condition-specific outcome measures. Methods A service evaluation was conducted over a 15-month period to assess our current treatment protocol used in the management of patients with atraumatic recurrent shoulder instability. The regime is reproducible with target-led progression milestones. Oxford Instability Shoulder Scores (OISS) and Western Ontario Shoulder Index (WOSI) scores were compared between baseline and final follow-up. Results Eighteen consecutive patients were included with mean follow-up of 4.5 months (range 1.35 months to 11.77 months). A statistically significant improvement was seen in both outcome measures. Mean OISS improved by 16.67 points (confidence interval: 12.34 to 20.99; p < 0.001). Mean WOSI improved by 36.76% (confidence interval: 28.46 to 45.06; p < 0.001). Conclusions For this small group of patients with recurrent atraumatic shoulder instability, the Derby Shoulder Instability Programme produced significant improvements over the short term, with a high level of patient compliance. This is the first study to include pathology-specific patient-reported outcome measures to assess outcomes from a specific and reproducible exercise regime in this group of patients. The findings support further research to evaluate the exercise protocol in a larger group of patients over the longer term.

  11. Wobble Pairs of the HDV Ribozyme Play Specific Roles in Stabilization of Active Site Dynamics

    PubMed Central

    Sripathi, Kamali N.; Banáš, Pavel; Reblova, Kamila; Šponer, Jiři; Otyepka, Michal

    2015-01-01

    The hepatitis delta virus (HDV) is the only known human pathogen whose genome contains a catalytic RNA motif (ribozyme). The overall architecture of the HDV ribozyme is that of a double-nested pseudoknot, with two GU pairs flanking the active site. Although extensive studies have shown that mutation of either wobble results in decreased catalytic activity, little work has focused on linking these mutations to specific structural effects on catalytic fitness. Here we use molecular dynamics simulations based on an activated structure to probe the active site dynamics as a result of wobble pair mutations. In both wild-type and mutant ribozymes, the in-line fitness of the active site (as a measure of catalytic proficiency) strongly depends on the presence of a C75(N3H3+)N1(O5′) hydrogen bond, which positions C75 as the general acid for the reaction. Our mutational analyses show that each GU wobble supports catalytically fit conformations in distinct ways; the reverse G25U20 wobble promotes high in-line fitness, high occupancy of the C75(N3H3+)G1(O5′) general-acid hydrogen bond and stabilization of the G1U37 wobble, while the G1U37 wobble acts more locally by stabilizing high in-line fitness and the C75(N3H3+)G1(O5′) hydrogen bond. We also find that stable type I A-minor and P1.1 hydrogen bonding above and below the active site, respectively, prevent local structural disorder from spreading and disrupting global conformation. Taken together, our results define specific, often redundant architectural roles for several structural motifs of the HDV ribozyme active site, expanding the known roles of these motifs within all HDV-like ribozymes and other structured RNAs. PMID:25631765

  12. Thermodynamic Activity Measurements with Knudsen Cell Mass Spectrometry

    NASA Technical Reports Server (NTRS)

    Copland, Evan H.; Jacobson, Nathan S.

    2001-01-01

    Coupling the Knudsen effusion method with mass spectrometry has proven to be one of the most useful experimental techniques for studying the equilibrium between condensed phases and complex vapors. The Knudsen effusion method involves placing a condensed sample in a Knudsen cell, a small "enclosure", that is uniformly heated and held until equilibrium is attained between the condensed and vapor phases. The vapor is continuously sampled by effusion through a small orifice in the cell. A molecular beam is formed from the effusing vapor and directed into a mass spectrometer for identification and pressure measurement of the species in the vapor phase. Knudsen cell mass spectrometry (KCMS) has been used for nearly fifty years now and continues to be a leading technique for obtaining thermodynamic data. Indeed, much of the well-established vapor specie data in the JANAF tables has been obtained from this technique. This is due to the extreme versatility of the technique. All classes of materials can be studied and all constituents of the vapor phase can be measured over a wide range of pressures (approximately 10(exp -4) to 10(exp -11) bar) and temperatures (500-2800 K). The ability to selectively measure different vapor species makes KCMS a very powerful tool for the measurement of component activities in metallic and ceramic solutions. Today several groups are applying KCMS to measure thermodynamic functions in multicomponent metallic and ceramic systems. Thermodynamic functions, especially component activities, are extremely important in the development of CALPHAD (Calculation of Phase Diagrams) type thermodynamic descriptions. These descriptions, in turn, are useful for modeling materials processing and predicting reactions such as oxide formation and fiber/matrix interactions. The leading experimental methods for measuring activities are the Galvanic cell or electro-motive force (EMF) technique and the KCMS technique. Each has specific advantages, depending on

  13. Context-specific behavioral surprise is differentially correlated with activity in anterior and posterior brain systems.

    PubMed

    Tobia, Michael J; Gläscher, Jan; Sommer, Tobias

    2016-06-15

    This experiment investigated whether behavioral surprise, an information-theoretic measure of the amount of memory and information integration associated with a response, is correlated with neural activity during decision making. A total of 30 participants (age 18-30) were scanned with functional MRI while completing 240 trials of a sequential decision-making task in which they selected an amount to wager from four possible values on each trial. Behavioral surprise was computed trial by trial using both context-free and context-specific formulations, and was used as a parametric modulator in functional MRI analyses. Whereas context-free surprise was not significantly correlated, two sets of clusters (P<0.005; cluster size>156 voxels) were differentially modulated by context-specific behavioral surprise. An anterior system comprised of the inferior frontal gyrus and anterior cingulate (each bilaterally), and left caudate, was positively modulated. A posterior system comprised of the posterior cingulate, parahippocampal gyrus and posterior hippocampus (each bilaterally), and left angular gyrus, was negatively modulated. These anticorrelated systems indicate that more surprising (resource demanding) actions recruit greater activity from the anterior system and less activity from the posterior system and less surprising actions (memory-guided) recruit greater activity from the posterior system and less activity from the anterior system. These results show that context-specific behavioral surprise is a unique neural signal and may be related to mechanisms for both cognitive control and memory-guided behavior, and support contemporary theories that the brain is a statistical observer of external and internal events. PMID:27110868

  14. Novel antimicrobial peptide specifically active against Porphyromonas gingivalis.

    PubMed

    Suwandecha, T; Srichana, T; Balekar, N; Nakpheng, T; Pangsomboon, K

    2015-09-01

    Porphyromonas gingivalis, the major etiologic agent of chronic periodontitis, produces a broad spectrum of virulence factors, including outer membrane vesicles, lipopolysaccharides, hemolysins and proteinases. Antimicrobial peptides (AMPs) including bacteriocins have been found to inhibit the growth of P. gingivalis; however, these peptides are relatively large molecules. Hence, it is difficult to synthesize them by a scale-up production. Therefore, this study aimed to synthesize a shorter AMP that was still active against P. gingivalis. A peptide that contained three cationic amino acids (Arg, His and Lys), two anionic amino acids (Glu and Asp), hydrophobic amino acids residues (Leu, Ile, Val, Ala and Pro) and hydrophilic residues (Ser and Gly) was obtained and named Pep-7. Its bioactivity and stability were tested after various treatments. The mechanism of action of Pep-7 and its toxicity to human red blood cells were investigated. The Pep-7 inhibited two pathogenic P. gingivalis ATCC 33277 and P. gingivalis ATCC 53978 (wp50) strains at a minimum bactericidal concentration (MBC) of 1.7 µM, but was ineffective against other oral microorganisms (P. intermedia, Tannerella forsythensis, Streptococcus salivarius and Streptococcus sanguinis). From transmission electron microscopy studies, Pep-7 caused pore formation at the poles of the cytoplasmic membranes of P. gingivalis. A concentration of Pep-7 at four times that of its MBC induced some hemolysis but only at 0.3%. The Pep-7 was heat stable under pressure (autoclave at 110 and 121 °C) and possessed activity over a pH range of 6.8-8.5. It was not toxic to periodontal cells over a range of 70.8-4.4 μM and did not induce toxic pro-inflammatory cytokines. The Pep-7 showed selective activity against Porphyromonas sp. by altering the permeability barriers of P. gingivalis. The Pep-7 was not mutagenic in vitro. This work highlighted the potential for the use of this synthetic Pep-7 against P. gingivalis. PMID:26041027

  15. Target cell-specific modulation of neuronal activity by astrocytes

    NASA Astrophysics Data System (ADS)

    Kozlov, A. S.; Angulo, M. C.; Audinat, E.; Charpak, S.

    2006-06-01

    Interaction between astrocytes and neurons enriches the behavior of brain circuits. By releasing glutamate and ATP, astrocytes can directly excite neurons and modulate synaptic transmission. In the rat olfactory bulb, we demonstrate that the release of GABA by astrocytes causes long-lasting and synchronous inhibition of mitral and granule cells. In addition, astrocytes release glutamate, leading to a selective activation of granule-cell NMDA receptors. Thus, by releasing excitatory and inhibitory neurotransmitters, astrocytes exert a complex modulatory control on the olfactory network. glutamate | GABA | inhibition | olfactory bulb | synchronization

  16. Active spectroscopic measurements using the ITER diagnostic system

    SciTech Connect

    Thomas, D. M.; Counsell, G.; Johnson, D.; Vasu, P.; Zvonkov, A.

    2010-10-15

    Active (beam-based) spectroscopic measurements are intended to provide a number of crucial parameters for the ITER device being built in Cadarache, France. These measurements include the determination of impurity ion temperatures, absolute densities, and velocity profiles, as well as the determination of the plasma current density profile. Because ITER will be the first experiment to study long timescale ({approx}1 h) fusion burn plasmas, of particular interest is the ability to study the profile of the thermalized helium ash resulting from the slowing down and confinement of the fusion alphas. These measurements will utilize both the 1 MeV heating neutral beams and a dedicated 100 keV hydrogen diagnostic neutral beam. A number of separate instruments are being designed and built by several of the ITER partners to meet the different spectroscopic measurement needs and to provide the maximum physics information. In this paper, we describe the planned measurements, the intended diagnostic ensemble, and we will discuss specific physics and engineering challenges for these measurements in ITER.

  17. Active spectroscopic measurements using the ITER diagnostic systema)

    NASA Astrophysics Data System (ADS)

    Thomas, D. M.; Counsell, G.; Johnson, D.; Vasu, P.; Zvonkov, A.

    2010-10-01

    Active (beam-based) spectroscopic measurements are intended to provide a number of crucial parameters for the ITER device being built in Cadarache, France. These measurements include the determination of impurity ion temperatures, absolute densities, and velocity profiles, as well as the determination of the plasma current density profile. Because ITER will be the first experiment to study long timescale (˜1 h) fusion burn plasmas, of particular interest is the ability to study the profile of the thermalized helium ash resulting from the slowing down and confinement of the fusion alphas. These measurements will utilize both the 1 MeV heating neutral beams and a dedicated 100 keV hydrogen diagnostic neutral beam. A number of separate instruments are being designed and built by several of the ITER partners to meet the different spectroscopic measurement needs and to provide the maximum physics information. In this paper, we describe the planned measurements, the intended diagnostic ensemble, and we will discuss specific physics and engineering challenges for these measurements in ITER.

  18. Are the correlates of active school transport context-specific?

    PubMed Central

    Larouche, R; Sarmiento, O L; Broyles, S T; Denstel, K D; Church, T S; Barreira, T V; Chaput, J-P; Fogelholm, M; Hu, G; Kuriyan, R; Kurpad, A; Lambert, E V; Maher, C; Maia, J; Matsudo, V; Olds, T; Onywera, V; Standage, M; Tremblay, M S; Tudor-Locke, C; Zhao, P; Katzmarzyk, P T

    2015-01-01

    OBJECTIVES: Previous research consistently indicates that children who engage in active school transport (AST) are more active than their peers who use motorized modes (car or bus). However, studies of the correlates of AST have been conducted predominantly in high-income countries and have yielded mixed findings. Using data from a heterogeneous sample of 12 country sites across the world, we investigated the correlates of AST in 9–11-year olds. METHODS: The analytical sample comprised 6555 children (53.8% girls), who reported their main travel mode to school and the duration of their school trip. Potential individual and neighborhood correlates of AST were assessed with a parent questionnaire adapted from previously validated instruments. Multilevel generalized linear mixed models (GLMM) were used to examine the associations between individual and neighborhood variables and the odds of engaging in AST while controlling for the child's school. Site moderated the relationship of seven of these variables with AST; therefore we present analyses stratified by site. RESULTS: The prevalence of AST varied from 5.2 to 79.4% across sites and the school-level intra-class correlation ranged from 0.00 to 0.56. For each site, the final GLMM included a different set of correlates of AST. Longer trip duration (that is, ⩾16 min versus ⩽15 min) was associated with lower odds of AST in eight sites. Other individual and neighborhood factors were associated with AST in three sites or less. CONCLUSIONS: Our results indicate wide variability in the prevalence and correlates of AST in a large sample of children from twelve geographically, economically and culturally diverse country sites. This suggests that AST interventions should not adopt a ‘one size fits all' approach. Future research should also explore the association between psychosocial factors and AST in different countries. PMID:27152191

  19. A METHOD FOR THE MEASUREMENT OF SITE-SPECIFIC TAUTOMERIC AND ZWITTERIONIC MICROSPECIES EQUILIBRIUM CONSTANTS

    EPA Science Inventory

    We describe a method for the individual measurement of simultaneously occurring, unimolecular, site-specific "microequilibrium" constants as in, for example, prototropic tautomerism and zwitterionic equilibria. Our method represents an elaboration of that of Nygren et al. (Anal. ...

  20. METHOD FOR THE MEASUREMENT OF SITE-SPECIFIC TAUTOMERIC AND ZWITTERIONIC MICROSPECIES EQUILIBRIUM CONSTANTS

    EPA Science Inventory

    We describe a method for the individual measurement of simultaneously occurring, unimolecular, site-specific “microequilibrium” constants as in, for example, prototropic tautomerism and zwitterionic equilibria. Our method represents an elaboration of that of Nygren et al. (Anal. ...

  1. 12 CFR 217.210 - Standardized measurement method for specific risk

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... deal-specific definitions of default; (B) Relevant information regarding the performance of the... measures of creditworthiness; average loan-to-value ratio; and industry and geographic diversification data... resecuritization positions, performance information on the underlying securitization exposures, for example,...

  2. Recommendations for Guidelines for Environment-Specific Magnetic-Field Measurements, Rapid Program Engineering Project #2

    SciTech Connect

    Electric Research and Management, Inc.; IIT Research Institute; Magnetic Measurements; Survey Research Center, University of California; T. Dan Bracken, Inc.

    1997-03-11

    The purpose of this project was to document widely applicable methods for characterizing the magnetic fields in a given environment, recognizing the many sources co-existing within that space. The guidelines are designed to allow the reader to follow an efficient process to (1) plan the goals and requirements of a magnetic-field study, (2) develop a study structure and protocol, and (3) document and carry out the plan. These guidelines take the reader first through the process of developing a basic study strategy, then through planning and performing the data collection. Last, the critical factors of data management, analysis reporting, and quality assurance are discussed. The guidelines are structured to allow the researcher to develop a protocol that responds to specific site and project needs. The Research and Public Information Dissemination Program (RAPID) is based on exposure to magnetic fields and the potential health effects. Therefore, the most important focus for these magnetic-field measurement guidelines is relevance to exposure. The assumed objective of an environment-specific measurement is to characterize the environment (given a set of occupants and magnetic-field sources) so that information about the exposure of the occupants may be inferred. Ideally, the researcher seeks to obtain complete or "perfect" information about these magnetic fields, so that personal exposure might also be modeled perfectly. However, complete data collection is not feasible. In fact, it has been made more difficult as the research field has moved to expand the list of field parameters measured, increasing the cost and complexity of performing a measurement and analyzing the data. The guidelines address this issue by guiding the user to design a measurement protocol that will gather the most exposure-relevant information based on the locations of people in relation to the sources. We suggest that the "microenvironment" become the base unit of area in a study, with

  3. A Comparison of a Subject-Specific and a General Measure of Critical Thinking

    ERIC Educational Resources Information Center

    Renaud, Robert D.; Murray, Harry G.

    2008-01-01

    The majority of studies reporting gains in college students' critical thinking due to instructional process variables measured critical thinking with subject-specific questions rather than general or decontextualized questions. However, it is uncertain whether these gains were attributable to the use of subject-specific questions or to other…

  4. Testing Time-Invariance of Variable Specificity in Repeated Measure Designs Using Structural Equation Modeling

    ERIC Educational Resources Information Center

    Raykov, Tenko; Amemiya, Yasuo

    2008-01-01

    A structural equation modeling method for examining time-invariance of variable specificity in longitudinal studies with multiple measures is outlined, which is developed within a confirmatory factor-analytic framework. The approach represents a likelihood ratio test for the hypothesis of stability in the specificity part of the residual term…

  5. Fluorescence of prostate-specific antigen as measured with a portable 1D scanner

    NASA Astrophysics Data System (ADS)

    Kim, Byeong C.; Jeong, Jin H.; Jeong, Dong S.; Kim, Young M.; Oh, Sang W.; Choi, Eui Y.; Kim, Jae H.; Nahm, Kie B.

    2005-01-01

    Prostate-specific antigen (PSA) is an androgen-dependent glycoprotein protease (M.W. 33 kDa) and a member of kallikrein super-family of serine protease, and has chymotrypsin-like enzymatic activity. It is synthesized by the prostate epithelial cells and found in the prostate gland and seminal plasma as a major protein. It is widely used as a clinical marker for diagnosis, screening, monitoring and prognosis of prostate cancer. In normal male adults, the concentration of PSA in the blood is below 4 ng/ml and this value increases in patients with the prostate cancer or the benign prostatic hyperplasia (BPH) due to its leakage into the circulatory system. As such, systematic monitoring of the PSA level in the blood can provide critical information about the progress of the prostatic disease. We have developed a compact integral system that can quantitatively measure the concentration of total PSA in human blood. This system utilizes the fluorescence emitted from the dye molecules attached to PSA molecules after appropriate immunoassay-based processing. Developed for the purpose of providing an affordable means of fast point-of-care testing of the prostate cancer, this system proved to be able to detect the presence of the PSA at the level of 0.18 ng/ml in less than 12 minutes, with the actual measurement taking less than 2 minutes. The design concept for this system is presented together with the result for a few representative samples.

  6. Methods to Measure Physical Activity Behaviors in Health Education Research

    ERIC Educational Resources Information Center

    Fitzhugh, Eugene C.

    2015-01-01

    Regular physical activity (PA) is an important concept to measure in health education research. The health education researcher might need to measure physical activity because it is the primary measure of interest, or PA might be a confounding measure that needs to be controlled for in statistical analysis. The purpose of this commentary is to…

  7. Measuring Physical Activity in Outdoor Community Recreational Environments: Implications for Research, Policy, and Practice

    PubMed Central

    Jones, Sydney A.; Stransky, Michelle; Evenson, Kelly R.

    2015-01-01

    Chronic diseases such as cardiovascular disease (CVD) are major contributors to escalating health care costs in the USA. Physical activity is an important protective factor against CVD, and the National Prevention Strategy recognizes active living (defined as a way of life that integrates physical activity into everyday routines) as a priority for improving the nation’s health. This paper focuses on developing more inclusive measures of physical activity in outdoor community recreational environments, specifically parks and trails, to enhance their usability for at-risk populations such as persons with mobility limitations. We develop an integrated conceptual framework for measuring physical activity in outdoor community recreational environments, describe examples of evidence-based tools for measuring physical activity in these settings, and discuss strategies to improve measurement of physical activity for persons with mobility limitations. Addressing these measurement issues is critically important to making progress towards national CVD goals pertaining to active community environments. PMID:26005510

  8. Pain Inhibition by Optogenetic Activation of Specific Anterior Cingulate Cortical Neurons

    PubMed Central

    Gu, Ling; Uhelski, Megan L.; Anand, Sanjay; Romero-Ortega, Mario; Kim, Young-tae; Fuchs, Perry N.; Mohanty, Samarendra K.

    2015-01-01

    Cumulative evidence from both humans and animals suggests that the anterior cingulate cortex (ACC) is important for pain-related perception, and thus a likely target for pain relief therapy. However, use of existing electrode based ACC stimulation has not significantly reduced pain, at least in part due to the lack of specificity and likely co-activation of both excitatory and inhibitory neurons. Herein, we report a dramatic reduction of pain behavior in transgenic mice by optogenetic stimulation of the inhibitory neural circuitry of the ACC expressing channelrhodopsin-2. Electrophysiological measurements confirmed that stimulation of ACC inhibitory neurons is associated with decreased neural activity in the ACC. Further, a distinct optogenetic stimulation intensity and frequency-dependent inhibition of spiking activity in the ACC was observed. Moreover, we confirmed specific electrophysiological responses from different neuronal units in the thalamus, in response to particular types of painful stimuli (i,e., formalin injection, pinch), which we found to be modulated by optogenetic control of the ACC inhibitory neurons. These results underscore the inhibition of the ACC as a clinical alternative in inhibiting chronic pain, and leads to a better understanding of the pain processing circuitry of the cingulate cortex. PMID:25714399

  9. The Amino Acid Specificity for Activation of Phenylalanine Hydroxylase Matches the Specificity for Stabilization of Regulatory Domain Dimers

    PubMed Central

    2016-01-01

    Liver phenylalanine hydroxylase is allosterically activated by phenylalanine. The structural changes that accompany activation have not been identified, but recent studies of the effects of phenylalanine on the isolated regulatory domain of the enzyme support a model in which phenylalanine binding promotes regulatory domain dimerization. Such a model predicts that compounds that stabilize the regulatory domain dimer will also activate the enzyme. Nuclear magnetic resonance spectroscopy and analytical ultracentrifugation were used to determine the ability of different amino acids and phenylalanine analogues to stabilize the regulatory domain dimer. The abilities of these compounds to activate the enzyme were analyzed by measuring their effects on the fluorescence change that accompanies activation and on the activity directly. At concentrations of 10–50 mM, d-phenylalanine, l-methionine, l-norleucine, and (S)-2-amino-3-phenyl-1-propanol were able to activate the enzyme to the same extent as 1 mM l-phenylalanine. Lower levels of activation were seen with l-4-aminophenylalanine, l-leucine, l-isoleucine, and 3-phenylpropionate. The ability of these compounds to stabilize the regulatory domain dimer agreed with their ability to activate the enzyme. These results support a model in which allosteric activation of phenylalanine hydroxylase is linked to dimerization of regulatory domains. PMID:26252467

  10. A Method to Measure Hydrolytic Activity of Adenosinetriphosphatases (ATPases)

    PubMed Central

    Bartolommei, Gianluca; Moncelli, Maria Rosa; Tadini-Buoninsegni, Francesco

    2013-01-01

    The detection of small amounts (nanomoles) of inorganic phosphate has a great interest in biochemistry. In particular, phosphate detection is useful to evaluate the rate of hydrolysis of phosphatases, that are enzymes able to remove phosphate from their substrate by hydrolytic cleavage. The hydrolysis rate is correlated to enzyme activity, an extremely important functional parameter. Among phosphatases there are the cation transporting adenosinetriphosphatases (ATPases), that produce inorganic phosphate by cleavage of the γ-phosphate of ATP. These membrane transporters have many fundamental physiological roles and are emerging as potential drug targets. ATPase hydrolytic activity is measured to test enzyme functionality, but it also provides useful information on possible inhibitory effects of molecules that interfere with the hydrolytic process. We have optimized a molybdenum-based protocol that makes use of potassium antimony (III) oxide tartrate (originally employed for phosphate detection in environmental analysis) to allow its use with phosphatase enzymes. In particular, the method was successfully applied to native and recombinant ATPases to demonstrate its reliability, validity, sensitivity and versatility. Our method introduces significant improvements to well-established experimental assays, which are currently employed for ATPase activity measurements. Therefore, it may be valuable in biochemical and biomedical investigations of ATPase enzymes, in combination with more specific tests, as well as in high throughput drug screening. PMID:23472215

  11. The Measurement of the Specific Latent Heat of Fusion of Ice: Two Improved Methods.

    ERIC Educational Resources Information Center

    Mak, S. Y.; Chun, C. K. W.

    2000-01-01

    Suggests two methods for measuring the specific latent heat of ice fusion for high school physics laboratories. The first method is an ice calorimeter which is made from simple materials. The second method improves the thermal contact and allows for a more accurate measurement. Lists instructions for both methods. (Author/YDS)

  12. Specifics of the hail parameter measurements using the optical precipitation gauge

    NASA Astrophysics Data System (ADS)

    Kalchikhin, V. V.; Kobzev, A. A.; Korolkov, V. A.; Tikhomirov, A. A.

    2015-11-01

    There are specifics of use of the obtaining and analyzing precipitation particle shadow images method for the hail precipitation investigations. Descriptions of the method and operation of the new optical rain gauge measuring system are presented. There are estimations of the device capabilities and prospects of its use for measurement of hail characteristics.

  13. Measuring motivation in schizophrenia: is a general state of motivation necessary for task-specific motivation?

    PubMed

    Choi, Jimmy; Choi, Kee-Hong; Felice Reddy, L; Fiszdon, Joanna M

    2014-03-01

    Despite the important role of motivation in rehabilitation and functional outcomes in schizophrenia, to date, there has been little emphasis on how motivation is assessed. This is important, since different measures may tap potentially discrete motivational constructs, which in turn may have very different associations to important outcomes. In the current study, we used baseline data from 71 schizophrenia spectrum outpatients enrolled in a rehabilitation program to examine the relationship between task-specific motivation, as measured by the Intrinsic Motivation Inventory (IMI), and a more general state of volition/initiation, as measured by the three item Quality of Life (QLS) motivation index. We also examined the relationship of these motivation measures to demographic, clinical and functional variables relevant to rehabilitation outcomes. The two motivation measures were not correlated, and participants with low general state motivation exhibited a full range of task-specific motivation. Only the QLS motivation index correlated with variables relevant to rehabilitation outcomes. The lack of associations between QLS motivation index and IMI subscales suggests that constructs tapped by these measures may be divergent in schizophrenia, and specifically that task-specific intrinsic motivation is not contingent on a general state of motivation. That is, even in individuals with a general low motivational state (i.e. amotivation), interventions aimed at increasing task-specific motivation may still be effective. Moreover, the pattern of interrelationships between the QLS motivation index and variables relevant to psychosocial rehabilitation supports its use in treatment outcome studies. PMID:24529609

  14. Measuring motivation in schizophrenia: Is a general state of motivation necessary for task-specific motivation?

    PubMed Central

    Choi, Jimmy; Choi, Kee-Hong; Reddy, Felice; Fiszdon, Joanna M.

    2014-01-01

    Despite the important role of motivation in rehabilitation and functional outcomes in schizophrenia, to date, there has been little emphasis on how motivation is assessed. This is important, since different measures may tap potentially discrete motivational constructs, which in turn may have very different associations to important outcomes. In the current study, we used baseline data from 71 schizophrenia spectrum outpatients enrolled in a rehabilitation program to examine the relationship between task-specific motivation, as measured by the Intrinsic Motivation Inventory (IMI), and a more general state of volition/initiation, as measured by the three item Quality of Life (QLS) motivation index. We also examined the relationship of these motivation measures to demographic, clinical and functional variables relevant to rehabilitation outcomes. The two motivation measures were not correlated, and participants with low general state motivation exhibited a full range of task-specific motivation. Only the QLS motivation index correlated with variables relevant to rehabilitation outcomes. The lack of associations between QLS motivation index and IMI subscales suggests that constructs tapped by these measures may be divergent in schizophrenia, and specifically that task-specific intrinsic motivation is not contingent on a general state of motivation. That is, even in individuals with a general low motivational state (i.e. amotivation), interventions aimed at increasing task-specific motivation may still be effective. Moreover, the pattern of interrelationships between the QLS motivation index and variables relevant to psychosocial rehabilitation supports its use in treatment outcome studies. PMID:24529609

  15. Evaluation and management of measurement uncertainty in the new generation Geometrical Product Specification (GPS)

    NASA Astrophysics Data System (ADS)

    Peng, Heping; Liu, Xiaojun; Jiang, Xiangqian; Xu, Zhengao

    2006-11-01

    Because of measurement errors objectively existing, measurement results always deviate from the "true value" of the measurand. Measurement uncertainty, as a quality index characterizing measurement results, attracts more attention worldwide. Its evaluation and expression will help understand measurement results, and promote international technical and commercial communication. According to the International Organization for Standardization ISO/BIPM "Guide to the Expression of Uncertainty in Measurement" - usually referred to as the GUM, this paper deals with the way of evaluation and expression of measurement uncertainties, describes the Procedure for Uncertainty of Measurement MAnagement (PUMA) in the new generation Geometrical Product Specification (GPS). An example of uncertainty evaluation with the PUMA method is given to illustrate the validity of PUMA. Since the concept of uncertainty in the new generation GPS has been expanded beyond just measurement uncertainty, the new way of evaluation and management of uncertainty throughout the whole GPS system remains to be developed.

  16. Measurement of optical activity of honey bee

    NASA Astrophysics Data System (ADS)

    Ortiz-Gutiérrez, Mauricio; Olivares-Pérez, Arturo; Salgado-Verduzco, Marco Antonio; Ibarra-Torres, Juan Carlos

    2016-03-01

    Optical activity of some substances, such as chiral molecules, often exhibits circular birefringence. Circular birefringence causes rotation of the vibration plane of the plane polarized light as it passes through the substance. In this work we present optical characterization of honey as function of the optical activity when it is placed in a polariscope that consists of a light source and properly arranged polarizing elements.

  17. PARTICULATE EMISSION MEASUREMENTS FROM CONTROLLED CONSTRUCTION ACTIVITIES

    EPA Science Inventory

    The report summarized the results of field testing of the effectiveness of control measures for sources of fugitive particulate emissions found at construction sites. The effectiveness of watering temporary, unpaved travel surfaces on emissions of particulate matter with aerodyna...

  18. Radioactivity of Potassium Solutions: A Comparison of Calculated Activity to Measured Activity from Gross Beta Counting and Gamma Spectroscopy

    SciTech Connect

    Gaylord, R F

    2005-07-26

    In order to determine if the measured beta activity for a solution containing potassium was exactly as predicted, particularly since the CES gas counter is not calibrated specifically with K-40, an experiment was conducted to compare measured activities from two radioanalytical methods (gamma spectroscopy and gas proportional counting) to calculated activities across a range of potassium concentrations. Potassium, being ubiquitous and naturally radioactive, is a well-known and common interference in gross beta counting methods. By measuring the observed beta activity due to K-40 in potassium-containing solutions across a wide range of concentrations, it was found that the observed beta activity agrees well with the beta activity calculated from the potassium concentration measured by standard inorganic analytical techniques, such as ICP-OES, and that using the measured potassium concentration to calculate the expected beta activity, and comparing this to the observed beta activity to determine if potassium can account for all the observed activity in a sample, is a valid technique. It was also observed that gamma spectroscopy is not an effective means of measuring K-40 activity below approximately 700 pCi/L, which corresponds to a solution with approximately 833 mg/L total potassium. Gas proportional counting for gross beta activity has a much lower detection limit, typically 20-50 picoCi/L for a liquid low in total dissolved solids, which corresponds to a potassium concentration of approximately 30-70 ppm K.

  19. Specific conductance measurements in central and western New York streams - A retrospective characterization

    USGS Publications Warehouse

    Kappel, William M.; Sinclair, Gaylen J.; Reddy, James E.; Eckhardt, David A.; deVries, M. Peter; Phillips, Margaret E.

    2012-01-01

    U.S. Geological Survey (USGS) Data Rescue Program funds were used to recover data from paper records for 139 streamgages across central and western New York State; 6,133 different streamflow measurement forms, collected between 1970-80, contained field water-quality measurements. The water-quality data were entered, reviewed, and uploaded into the USGS National Water Information System. In total, 4,285 unique site visits were added to the database. The new values represent baseline water quality from which to measure change and will lead to a comparison of water-quality change over the last 40 years and into the future. Specific conductance was one of the measured properties and represents a simple way to determine if ambient inorganic water quality has been altered by anthropogenic (road salt runoff, wastewater discharges, or natural gas development) or natural sources. The objective of this report is to describe ambient specific conductance characteristics of surface water across the central and western part of New York. This report presents median specific conductance of stream discharge for the period 1970-80 and a description of the relation between specific conductance and concentrations of total dissolved solids (TDS) retrieved from the USGS National Water Information System (NWIS) database from 1955 to present. The data descriptions provide a baseline of surface-water specific conductance data that can used for comparison to current and future measurements in New York streams.

  20. How Does Attention Relate to the Ability-Specific and Position-Specific Components of Reasoning Measured by APM?

    ERIC Educational Resources Information Center

    Ren, Xuezhu; Goldhammer, Frank; Moosbrugger, Helfried; Schweizer, Karl

    2012-01-01

    The aim of the present study was to clarify the nature of the ability-specific and position-specific components of Raven's Advanced Progressive Matrices (APM) by relating them to a number of types of attention. The ability-specific component represents the constant part of cognitive performance whereas the position-specific component reflects the…

  1. Are Preschool Children Active Enough? Objectively Measured Physical Activity Levels

    ERIC Educational Resources Information Center

    Cardon, Greet M.; De Bourdeaudhuij, Ilse M. M.

    2008-01-01

    The present study aimed to describe accelerometer-based physical activity levels in 4- and 5-year-old children (N = 76) on 2 weekdays and 2 weekend days. The children were sedentary for 9.6 hr (85%) daily, while they engaged in moderate to vigorous physical activity (MVPA) for 34 min (5%). Only 7% of the children engaged in MVPA for 60 min per…

  2. Peptide biosensors for the electrochemical measurement of protein kinase activity.

    PubMed

    Kerman, Kagan; Song, Haifeng; Duncan, James S; Litchfield, David W; Kraatz, Heinz-Bernhard

    2008-12-15

    The kinase activities are elucidated using the novel redox-active cosubstrate adenosine 5'-[gamma-ferrocene] triphosphate (Fc-ATP), which enables the kinase-catalyzed transfer of a redox active gamma-phosphate-Fc to a hydroxyamino acid. In this report, a versatile electrochemical biosensor is developed for monitoring the activity and inhibition of a serine/threonine kinase, casein kinase 2 (CK2), and protein tyrosine kinases, Abl1-T315I and HER2, in buffered solutions and in cell lysates. The method is based on the labeling of a specific phosphorylation event with Fc, followed by electrochemical detection. The electrochemical response obtained from the "ferrocenylated" peptides enables monitoring the activity of the kinase and its substrate, as well as the inhibition of small molecule inhibitors on protein phosphorylation. Kinetic information was extracted from the electrochemical measurements for the determination of K(m) and V(m) values, which were in agreement with those previously reported. Kinase reactions were also performed in the presence of well-defined inhibitors of CK2, 4,5,6,7-tetrabromo-2-azabenzimidazole, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole, and E-3-(2,3,4,5-tetrabromophenyl)acrylic acid as well as the nonspecific kinase inhibitors, staurosporine and N-benzoylstaurosporine. On the basis of the dependency of the Fc signal on inhibitor concentration, K(i) of the inhibitors was estimated, which were also in agreement with the literature values. The performance of the biosensor was optimized including the kinase reaction, incubation with Fc-ATP, and the small molecule inhibitors. Peptide modified electrochemical biosensors are promising candidates for cost-effective in vitro kinase activity and inhibitor screening assays. PMID:18989981

  3. Effects of active vs. passive recovery on repeated rugby-specific exercises.

    PubMed

    Jougla, A; Micallef, J P; Mottet, D

    2010-05-01

    The aim of this study was to determine the effects of active vs. passive recovery on performance of a rugby-specific intermittent test in rugby union players. Seven male rugby players (20.6+/-0.5 yrs; 181.9+/-10.0 cm; 94.5+/-12.8 kg) performed in random order, over two separate sessions, a specific repeated-sprint rugby test, the Narbonne test (6 x 4 consecutive actions: 1, scrummaging; 2, agility sprinting; 3, tackling; 4, straight sprinting) with 30s of passive or active recovery (running at 50% of maximal aerobic speed). The Narbonne tests were completed before (pre-test) and after (post-test) a 30-min rugby match. During the Narbonne test, scrum forces, agility and sprint times, heart rate and rate of perceived exertion were measured. Scrum forces were lower in active (74.9+/-13.4 kg) than in passive recovery (90.4+/-20.9 kg), only during the post-test (p<0.05). Fatigue index (%) (p<0.05) and total sprint time (s) (p<0.01) were significantly greater in active than in passive recovery, both during the pre-test (11.5+/-5.7% vs. 6.7+/-4.5% and 18.1+/-1.3s vs. 16.9+/-0.9s) and the post-test (7.3+/-3.3% vs. 4.3+/-1.5% and 18.3+/-1.6s vs. 16.9+/-1.1s). Consequently, the results indicated that passive recovery enabled better performance during the Narbonne test. However, it is obviously impractical to suggest that players should stand still during and following repeated-sprint bouts: the players have to move to ensure they have taken an optimal position. PMID:19560972

  4. MEASURING CHOLINESTERASE ACTIVITY IN HUMAN SALIVA

    EPA Science Inventory

    To assess the potential for using saliva in pesticide biomonitoring, the consistency of cholinesterase activity in human saliva collected over time was examined. In this pilot study, saliva was collected from 20 healthy adults once per week for 5 consecutive weeks using 2 differe...

  5. MEASURING CHOLINESTERASE ACTIVITY IN HUMAN SALIVA.

    EPA Science Inventory

    To assess the potential for using saliva in pesticide biomonitoring, the consistency of cholinesterase activity in human saliva collected over time was examined. In this pilot study, saliva was collected from 20 healthy adults once per week for 5 consecutive weeks using 2 differe...

  6. The protein structures that shape caspase activity, specificity, activation and inhibition

    PubMed Central

    Fuentes-Prior, Pablo; Salvesen, Guy S.

    2004-01-01

    The death morphology commonly known as apoptosis results from a post-translational pathway driven largely by specific limited proteolysis. In the last decade the structural basis for apoptosis regulation has moved from nothing to ‘quite good’, and we now know the fundamental structures of examples from the initiator phase, the pre-mitochondrial regulator phase, the executioner phase, inhibitors and their antagonists, and even the structures of some substrates. The field is as well advanced as the best known of proteolytic pathways, the coagulation cascade. Fundamentally new mechanisms in protease regulation have been disclosed. Structural evidence suggests that caspases have an unusual catalytic mechanism, and that they are activated by apparently unrelated events, depending on which position in the apoptotic pathway they occupy. Some naturally occurring caspase inhibitors have adopted classic inhibition strategies, but other have revealed completely novel mechanisms. All of the structural and mechanistic information can, and is, being applied to drive therapeutic strategies to combat overactivation of apoptosis in degenerative disease, and underactivation in neoplasia. We present a comprehensive review of the caspases, their regulators and inhibitors from a structural and mechanistic point of view, and with an aim to consolidate the many threads that define the rapid growth of this field. PMID:15450003

  7. The National Neurosurgery Quality and Outcomes Database Qualified Clinical Data Registry: 2015 measure specifications and rationale.

    PubMed

    Parker, Scott L; McGirt, Matthew J; Bekelis, Kimon; Holland, Christopher M; Davies, Jason; Devin, Clinton J; Atkins, Tyler; Knightly, Jack; Groman, Rachel; Zyung, Irene; Asher, Anthony L

    2015-12-01

    Meaningful quality measurement and public reporting have the potential to facilitate targeted outcome improvement, practice-based learning, shared decision making, and effective resource utilization. Recent developments in national quality reporting programs, such as the Centers for Medicare & Medicaid Services Qualified Clinical Data Registry (QCDR) reporting option, have enhanced the ability of specialty groups to develop relevant quality measures of the care they deliver. QCDRs will complete the collection and submission of Physician Quality Reporting System (PQRS) quality measures data on behalf of individual eligible professionals. The National Neurosurgery Quality and Outcomes Database (N(2)QOD) offers 21 non-PQRS measures, initially focused on spine procedures, which are the first specialty-specific measures for neurosurgery. Securing QCDR status for N(2)QOD is a tremendously important accomplishment for our specialty. This program will ensure that data collected through our registries and used for PQRS is meaningful for neurosurgeons, related spine care practitioners, their patients, and other stakeholders. The 2015 N(2)QOD QCDR is further evidence of neurosurgery's commitment to substantively advancing the health care quality paradigm. The following manuscript outlines the measures now approved for use in the 2015 N(2)QOD QCDR. Measure specifications (measure type and descriptions, related measures, if any, as well as relevant National Quality Strategy domain[s]) along with rationale are provided for each measure. PMID:26621418

  8. Performance on indirect measures of race evaluation predicts amygdala activation.

    PubMed

    Phelps, E A; O'Connor, K J; Cunningham, W A; Funayama, E S; Gatenby, J C; Gore, J C; Banaji, M R

    2000-09-01

    We used fMRI to explore the neural substrates involved in the unconscious evaluation of Black and White social groups. Specifically, we focused on the amygdala, a subcortical structure known to play a role in emotional learning and evaluation. In Experiment 1, White American subjects observed faces of unfamiliar Black and White males. The strength of amygdala activation to Black-versus-White faces was correlated with two indirect (unconscious) measures of race evaluation (Implicit Association Test [IAT] and potentiated startle), but not with the direct (conscious) expression of race attitudes. In Experiment 2, these patterns were not obtained when the stimulus faces belonged to familiar and positively regarded Black and White individuals. Together, these results suggest that amygdala and behavioral responses to Black-versus-White faces in White subjects reflect cultural evaluations of social groups modified by individual experience. PMID:11054916

  9. Low temperature specific heat measurement of a carbon-carbon composite

    NASA Astrophysics Data System (ADS)

    Yang, G.; Migone, A. D.

    A.c. calorimetry was used to determine the value of the specific heat at low temperatures for a sample of Textron's fine weave pierced fabric (FWPF) composite, a three-directional carbon-carbon composite with orthogonal fibre orientation. The heat capacity measurements were performed from 30 to 100 K at nearly 1 K intervals. The specific heat values for the composite are similar to those reported in the literature for graphite in the same temperature range.

  10. Structural basis of specific inhibition of tissue-type plasminogen activator by plasminogen activators inhibitor-1

    PubMed Central

    Gong, Lihu; Liu, Min; Zeng, Tu; Shi, Xiaoli; Yuan, Cai; Andreasen, Peter A.; Huang, Mingdong

    2016-01-01

    Thrombosis is a leading cause of death worldwide [1]. Recombinant tissue-type plasminogen activator (tPA) is the FDA-approved thrombolytic drug for ischemic strokes, myocardial infarction and pulmonary embolism. tPA is a multi-domain serine protease of the trypsin-family [2] and catalyses the critical step in fibrinolysis [3], converting the zymogen plasminogen to the active serine protease plasmin, which degrades the fibrin network of thrombi and blood clots. tPA is rapidly inactivated by endogenous plasminogen activators inhibitor-1 (PAI-1) [4] (Fig. 1). Engineering on tPA to reduce its inhibition by PAI-1 without compromising its thrombolytic effect is a continuous effort [5]. Tenecteplase (TNK-tPA) is a newer generation of tPA variant showing slower inhibition by PAI-1 [6]. Extensive studies to understand the molecular interactions between tPA and PAI-1 have been carried out [7], [8], [9], [10], [11], [12], [13], [14], [15], [16], [17], [18], however, the precise details at atomic resolution remain unknown. We report the crystal structure of tPA·PAI-1 complex here. The methods required to achieve these data include: (1) recombinant expression and purification of a PAI-1 variant (14-1B) containing four mutations (N150H, K154T, Q319L, and M354I), and a tPA serine protease domain (tPA-SPD) variant with three mutations (C122A, N173Q, and S195A, in the chymotrypsin numbering) [19]; (2) formation of a tPA-SPD·PAI-1 Michaëlis complex in vitro [19]; and (3) solving the three-dimensional structure for this complex by X-ray crystallography [deposited in the PDB database as 5BRR]. The data explain the specificity of PAI-1 for tPA and uPA [19], [20], and provide structural basis to design newer generation of PAI-1-resistant tPA variants as thrombolytic agents [19]. PMID:26909366

  11. Structural basis of specific inhibition of tissue-type plasminogen activator by plasminogen activators inhibitor-1.

    PubMed

    Gong, Lihu; Liu, Min; Zeng, Tu; Shi, Xiaoli; Yuan, Cai; Andreasen, Peter A; Huang, Mingdong

    2016-03-01

    Thrombosis is a leading cause of death worldwide [1]. Recombinant tissue-type plasminogen activator (tPA) is the FDA-approved thrombolytic drug for ischemic strokes, myocardial infarction and pulmonary embolism. tPA is a multi-domain serine protease of the trypsin-family [2] and catalyses the critical step in fibrinolysis [3], converting the zymogen plasminogen to the active serine protease plasmin, which degrades the fibrin network of thrombi and blood clots. tPA is rapidly inactivated by endogenous plasminogen activators inhibitor-1 (PAI-1) [4] (Fig. 1). Engineering on tPA to reduce its inhibition by PAI-1 without compromising its thrombolytic effect is a continuous effort [5]. Tenecteplase (TNK-tPA) is a newer generation of tPA variant showing slower inhibition by PAI-1 [6]. Extensive studies to understand the molecular interactions between tPA and PAI-1 have been carried out [7], [8], [9], [10], [11], [12], [13], [14], [15], [16], [17], [18], however, the precise details at atomic resolution remain unknown. We report the crystal structure of tPA·PAI-1 complex here. The methods required to achieve these data include: (1) recombinant expression and purification of a PAI-1 variant (14-1B) containing four mutations (N150H, K154T, Q319L, and M354I), and a tPA serine protease domain (tPA-SPD) variant with three mutations (C122A, N173Q, and S195A, in the chymotrypsin numbering) [19]; (2) formation of a tPA-SPD·PAI-1 Michaëlis complex in vitro [19]; and (3) solving the three-dimensional structure for this complex by X-ray crystallography [deposited in the PDB database as 5BRR]. The data explain the specificity of PAI-1 for tPA and uPA [19], [20], and provide structural basis to design newer generation of PAI-1-resistant tPA variants as thrombolytic agents [19]. PMID:26909366

  12. Sensitivity, specificity, and predictive values of a forearm blood pressure measurement method in severe obesity.

    PubMed

    Leblanc, Marie-Ève; Cloutier, Lyne; Poirier, Paul

    2015-04-01

    For blood pressure assessment, it has been reported that forearm blood pressure measurement appears to be as valid as an upper-arm blood pressure measurement method in individuals with severe obesity when correlated to the intra-arterial method, considered as the gold standard. Sensitivity, specificity, and predictive values for the presence of systemic hypertension in 25 severely obese patients from 352 blood pressure measurements were calculated. The sensitivity (0.98) and the positive predictive value (0.93) for hypertension on forearm blood pressure measurements are excellent, indicating that the forearm approach is a promising alternative to systemic hypertension diagnosis in severe obesity. PMID:25757220

  13. Crystal structures of human tissue kallikrein 4: activity modulation by a specific zinc binding site.

    PubMed

    Debela, Mekdes; Magdolen, Viktor; Grimminger, Valerie; Sommerhoff, Christian; Messerschmidt, Albrecht; Huber, Robert; Friedrich, Rainer; Bode, Wolfram; Goettig, Peter

    2006-10-01

    Human tissue kallikrein 4 (hK4) belongs to a 15-member family of closely related serine proteinases. hK4 is predominantly expressed in prostate, activates hK3/PSA, and is up-regulated in prostate and ovarian cancer. We have identified active monomers of recombinant hK4 besides inactive oligomers in solution. hK4 crystallised in the presence of zinc, nickel, and cobalt ions in three crystal forms containing cyclic tetramers and octamers. These structures display a novel metal site between His25 and Glu77 that links the 70-80 loop with the N-terminal segment. Micromolar zinc as present in prostatic fluid inhibits the enzymatic activity of hK4 against fluorogenic substrates. In our measurements, wild-type hK4 exhibited a zinc inhibition constant (IC50) of 16 microM including a permanent residual activity, in contrast to the zinc-independent mutants H25A and E77A. Since the Ile16 N terminus of wild-type hK4 becomes more accessible for acetylating agents in the presence of zinc, we propose that zinc affects the hK4 active site via the salt-bridge formed between the N terminus and Asp194 required for a functional active site. hK4 possesses an unusual 99-loop that creates a groove-like acidic S2 subsite. These findings explain the observed specificity of hK4 for the P1 to P4 substrate residues. Moreover, hK4 shows a negatively charged surface patch, which may represent an exosite for prime-side substrate recognition. PMID:16950394

  14. Mass specific optical absorption coefficients of mineral dust components measured by a multi wavelength photoacoustic spectrometer

    NASA Astrophysics Data System (ADS)

    Utry, N.; Ajtai, T.; Pintér, M.; Tombácz, E.; Illés, E.; Bozóki, Z.; Szabó, G.

    2014-09-01

    Mass specific optical absorption coefficients of various mineral dust components including silicate clays (illite, kaolin and bentonite), oxides (quartz, hematite and rutile), and carbonate (limestone) were determined at wavelengths of 1064, 532, 355 and 266 nm. These values were calculated from aerosol optical absorption coefficients measured by a multi-wavelength photoacoustic (PA) instrument, the mass concentration and the number size distribution of the generated aerosol samples as well as the size transfer functions of the measuring instruments. These results are expected to have considerable importance in global radiative forcing calculations. They can also serve as reference for validating calculated wavelength dependent imaginary parts (κ) of complex refractive indices which up to now have been typically deduced from bulk phase measurements by using indirect measurement methods. Accordingly, the presented comparison of the measured and calculated aerosol optical absorption spectra revealed the strong need for standardized sample preparation and measurement methodology in case of bulk phase measurements.

  15. Effect of source depth on the specificity of bipolar EEG measurements.

    PubMed

    Ryynanen, Outi; Vaisanen, Juho; Hyttinen, Jari; Malmivuo, Jaakko

    2006-01-01

    The purpose of the present study was to evaluate how the brain sources located at different depths can be most effectively measured with bipolar EEG leads. The specificity of an EEG lead to detect sources was studied with a new parameter called region of interest sensitivity ratio (ROISR) by employing a spherical head model. We studied the specificity as a function of electrode distance and further as a function of scalp:skull:brain resistivity ratio. The simulations indicate that the closer to the surface of the brain the source is located, the shorter is the interelectrode distance in the optimal lead. Also in the case of superficial sources, the small misplacement of the electrodes results in a substantial decrease in specificity. The resistivity ratio has the largest effect on the specificity, when the source is located close to the surface of the brain. However in the case of deep sources, the resistivity ratio has only minimal effect on the specificity. PMID:17945620

  16. Accuracy of Computational Cerebral Aneurysm Hemodynamics Using Patient-Specific Endovascular Measurements

    PubMed Central

    McGah, Patrick M.; Levitt, Michael R.; Barbour, Michael C.; Morton, Ryan P.; Nerva, John D.; Mourad, Pierre D.; Ghodke, Basavaraj V.; Hallam, Danial K.; Sekhar, Laligam N.; Kim, Louis J.; Aliseda, Alberto

    2013-01-01

    Computational hemodynamic simulations of cerebral aneurysms have traditionally relied on stereotypical boundary conditions (such as blood flow velocity and blood pressure) derived from published values as patient-specific measurements are unavailable or difficult to collect. However, controversy persists over the necessity of incorporating such patient specific conditions into computational analyses. We perform simulations using both endovascular-derived patient-specific and typical literature-derived inflow and outflow boundary conditions. Detailed three-dimensional anatomical models of the cerebral vasculature are developed from rotational angiography data, and blood flow velocity and pressure are measured in situ by a dual-sensor pressure and velocity endovascular guidewire at multiple peri-aneurysmal locations in ten unruptured cerebral aneurysms. These measurements are used to define inflow and outflow boundary conditions for computational hemodynamic models of the aneurysms. The additional in situ measurements which are not prescribed in the simulation are then used to assess the accuracy of the simulated flow velocity and pressure drop. Simulated velocities using patient-specific boundary conditions show good agreement with the guidewire measurements at measurement locations inside the domain, with no bias in the agreement and a random scatter of ≈25%. Simulated velocities using the simplified, literature-derived values show a systematic bias and over-predicted velocity by ≈30% with a random scatter of ≈40%. Computational hemodynamics using endovascularly measured patient-specific boundary conditions have the potential to improve treatment predictions as they provide more accurate and precise results of the aneurysmal hemodynamics than those based on commonly accepted reference values for boundary conditions. PMID:24162859

  17. The effect of trypsin and chymotrypsin on the bactericidal activity and specific antibody activity of bovine colostrum.

    PubMed Central

    Brock, J H; Arzabe, R; Piñeiro, A; Olivito, A M

    1977-01-01

    Digestion of bovine colostral whey with trypsin or chymotrypsin caused a progressive loss of the complement-mediated bactericidal activity of naturally-occurring colostral antibodies of E. coli 0111. Bactericidal activity was associated primarily with IgG1 immunoglobulin and to a lesser extent with IgM. Chymotrypsin preferentially attacked IgM, destroying its antibacterial activity and producing an apparent decrease in its mol wt. Trypsin preferentially attacked IgG1, but loss of antibacterial activity was in this case not accompanied by a decrease in molecular weight. Using colostral whey with antiperoxidase activity it was shown that the kinetics of loss of specific antibody activity were similar to those of loss of bactericidal activity. It is therefore suggested that trypsin may cause a loss of specific antibody activity of colostral IgG1 without cleaving the immunoglobulin molecule. PMID:321342

  18. Towards Fast In-line Measurement of Water Activity

    NASA Astrophysics Data System (ADS)

    Nielsen, J.; Andreasen, M. B.; Pedersen, M.; Rasmussen, M. K.

    2015-03-01

    Water activity is widely used as a key parameter in controlling the quality of food and feed products, among others. For determining the water activity, the material is sampled from the manufacturing process and measured in the laboratory with water activity analyzers. The sampling procedure can lead to non-representative measurements, the measurement process is time consuming, and much of the produced material may be wasted before the measurement results are available. To reduce waste and to be able to optimize production processes, industry requires in-line measurement of relevant quality determining parameters, hereunder the water activity. In cooperation with a manufacturer of systems for automatic in-line sampling and measurement of moisture, density, and the size of items, a project was defined to also enable the manufacturer's existing products to perform automatic measurement of the water activity in a sample. The aim was to develop a measurement system with the ability to operate in an industrial environment, which in the end would increase the measurement speed significantly and minimize the problems related to the handling of samples. In the paper the selection and characterization of the sensors, the design of a measurement chamber, and various issues of modeling and methods to reduce measurement time are discussed. The paper also presents water activity measurements obtained from food and feed products with the system, and shows that reliable results can be obtained in a few minutes with a proper design of the measurement chamber and selection of a model.

  19. Specific heat and electrical resistivity of niobium measured by subsecond calorimetric technique

    NASA Astrophysics Data System (ADS)

    Maglić, K. D.; Perović, N. Lj.; Vuković, G. S.; Zeković, Lj. P.

    1994-09-01

    This paper presents results of measurements of specific heat and electrical resistivity of niobium from ambient temperature to the experimental limit of the equipment which is close to 2500 K. The study used a contact thermometry variant of the millisecond resolution pulse calorimetry developed at the Institute of Nuclear Sciences VINČA. In the experiments exceeding 1000 K, thermocouple thermometry was supplemented with parallel pyrometric temperature measurements. This, together with application of tungsten; rhenium thermocouple thermometry, increased the temperature range of measurements to 2500 K. In the range where two thermometries overlap, data on the specimen emittance were also generated. Novelties in the method, the results on electrical resistivity. specific heat, hemispherical total emittance and normal spectral emittance of niobium, and accuracies attained in different property measurements are discussed.

  20. The Activity Chain Safety and Liveness Specification of Composite Web Services

    NASA Astrophysics Data System (ADS)

    Chen, Bo; Huang, Xiaomei

    Web service composition is most impressing method for development and deployment of e-business. Description and modeling the behavior requirements of composite Web services for users and verifying composite Web service compliance to specific requirements is an important key in design of services. But most work does not address the issue of how to model the requirements that the BPEL4WS processes are supposed to satisfy. The specifications in verification works are general temporal relation based on activity or scenario in essence. Distinguish with these work, we propose a novel concept of behavior specification based on activity chain in which granularity is between activity and scenario. Chain existence mode, chain absence mode are designed to express such behavioral requirements based on activity chain that is similar with safety or liveness specification based on activity respectively. Encode them on Labeled Transition System LTS and then give them exact operation semantics. Finally, an example is illustrated.

  1. Highly sensitive assay for acetylcholinesterase activity and inhibition based on a specifically reactive photonic nanostructure.

    PubMed

    Tian, Tian; Li, Xuesong; Cui, Jiecheng; Li, Jian; Lan, Yue; Wang, Chen; Zhang, Meng; Wang, Hui; Li, Guangtao

    2014-09-10

    Assays for acetylcholinesterase (AChE) with high sensitivity and high selectivity as well as facile manipulation have been urgently required in various fields. In this work, a reaction-based photonic strategy was developed for the efficient assay of AChE activity and inhibition based on the synergetic combination of the specific thiol-maleimide addition reaction with photonic porous structure. It was found that various applications including detection of AChE activity, measurement of the related enzymatic kinetics, and screening of inhibitors could be efficiently implemented using such strategy. Remarkably, the unique photonic nanostructure endows the constructed sensing platform with high sensitivity with a limit of detection (LOD) of 5 mU/mL for AChE activity, high selectivity, and self-reporting signaling. Moreover, the label-free solid film-based sensing approach described here has advantages of facile manipulation and bare-eye readout, compared with conventional liquid-phase methods, exhibiting promising potential in practical application for the AChE assay. PMID:25130420

  2. Active specific immunotherapy using the immune reaction of a low-dose irradiated tumor tissue. [Mice

    SciTech Connect

    Ogawa, Y.; Imanaka, K.; Ashida, C.; Takashima, H.; Imajo, Y.; Kimura, S.

    1983-04-01

    Active specific immunotherapy using the immune reaction of a low-dose irradiated tumor tissue was studied on the transplanted MM46 tumor of female C3H/He mice after radiotherapy. MM46 tumor cells were inoculated into the right hind paws of mice. On the 5th day, irradiation with the dose irradiated tumor tissue (2000 rad on the fifth day), were injected into the left hind paws of the tumor-bearing mice. Effectiveness of this active specific immunotherapy against tumor was evaluated by the regression of tumor and survival rate of mice. Tumor was markedly regressed and survival rate was significantly increased by the active specific immunitherapy.

  3. Linkage Specificity and Role of Properdin in Activation of the Alternative Complement Pathway by Fungal Glycans

    PubMed Central

    Agarwal, Sarika; Specht, Charles A.; Huang, Haibin; Ostroff, Gary R.; Ram, Sanjay; Rice, Peter A.; Levitz, Stuart M.

    2011-01-01

    ABSTRACT Fungal cell walls are predominantly composed of glucans, mannans, and chitin. Recognition of these glycans by the innate immune system is a critical component of host defenses against the mycoses. Complement, an important arm of innate immunity, plays a significant role in fungal pathogenesis, especially the alternative pathway (AP). Here we determine that the glycan monosaccharide composition and glycosidic linkages affect AP activation and C3 deposition. Furthermore, properdin, a positive regulator of the AP, contributes to these functions. AP activation by glycan particles that varied in composition and linkage was measured by C3a generation in serum treated with 10 mM EGTA and 10 mM Mg2+ (Mg-EGTA-treated serum) (AP specific; properdin functional) or Mg-EGTA-treated serum that lacked functional properdin. Particles that contained either β1→3 or β1→6 glucans or both generated large and similar amounts of C3a when the AP was intact. Blocking properdin function resulted in 5- to 10-fold-less C3a production by particulate β1→3 glucans. However, particulate β1→6 glucans generated C3a via the AP only in the presence of intact properdin. Interestingly, zymosan and glucan-mannan particles (GMP), which contain both β-glucans and mannans, also required properdin to generate C3a. The β1→4 glycans chitin and chitosan minimally activated C3 even when properdin was functional. Finally, properdin binding to glucan particles (GP) and zymosan in serum required active C3. Properdin colocalized with bound C3, suggesting that in the presence of serum, properdin bound indirectly to glycans through C3 convertases. These findings provide a better understanding of how properdin facilitates AP activation by fungi through interaction with the cell wall components. PMID:21878570

  4. Phosphate oxygen isotope ratio proxy for specific microbial activity in marine sediments (Peru Margin)

    NASA Astrophysics Data System (ADS)

    Liang, Y.; Blake, R. E.

    2005-12-01

    Oxygen (O) isotope ratios of biogenic apatites have been widely used as paleotemperature and environmental geochemical proxies. With improved knowledge of the phosphate O isotope effects of different P cycling pathways, the δ18O value of inorganic phosphate (δ18OP) has been proposed as a useful proxy and tracer of biological reactions and P cycling in natural environments[1,2,3,4]. Being the only way of removing P from oceanic water, sedimentary P burial is one of the most important processes during biogeochemical cycling of P. The high concentrations of organic matter and pronounced microbial activity at ODP Site 1230 along the Peru Margin result in unusually high interstitial water phosphate concentrations, which provides a unique opportunity to use δ18OP to investigate inorganic phosphate (Pi) regeneration and P cycling pathways in marine sediments. The isotopic measurements of both dissolved inorganic phosphate (DIP) and bulk sediment Pi show that DIP δ18OP values are affected by three different processes, which are all induced by specific microbial activities present in the sediments. In sediments at ~ 65 to 120 mbsf, porewater DIP is derived from dissolved organophosphorus compounds (DOP) through enzymatic degradation pathways, evidenced by both DIP δ18OP values and interstitial water chemistry. Measured porewater DIP δ18OP values also suggest that 4 to 8% of interstitial water DIP reflects regeneration of Pi from Porg by microbially-synthesized enzymes. Throughout the sediment column and especially at ~ 120 to 150 mbsf, DIP is released from the sediments by microbially-induced reductive dissolution of Fe-oxides, which contributes to the overall high DIP concentrations at Site 1230. The third and dominant process controlling measured DIP δ18OP values is microbial turnover of regenerated Pi. The presence of high microbial activities in organic-rich Site 1230 sediments promotes the remobilization of P and affects marine P cycling by potentially enhancing

  5. Neutron Yield Measurements via Aluminum Activation

    SciTech Connect

    1999-12-08

    Neutron activation of aluminum may occur by several neutron capture reactions. Four such reactions are described here: {sup 27}Al + n = {sup 28}Al, {sup 27}Al(n,{alpha}){sup 24}Na, {sup 27}Al(n, 2n){sup 26}Al and {sup 27}Al(n,p){sup 27}Mg. The radioactive nuclei {sup 28}Al, {sup 24}Na, and {sup 27}Mg, which are produced via the {sup 27}Al + n = {sup 28}Al, {sup 27}Al(n,{alpha}){sup 24}Na and {sup 27}Al(n,p){sup 27}Mg neutron reactions, beta decay to excited states of {sup 28}Si, {sup 24}Mg and {sup 27}Al respectively. These excited states then emit gamma rays as the nuclei de-excite to their respective ground states.

  6. Interpretation of Spectrometric Measurements of Active Geostationary Satellites

    NASA Astrophysics Data System (ADS)

    Bedard, D.; Wade, G.

    2014-09-01

    Over 5000 visible near-infrared (VNIR) spectrometric measurements of active geostationary satellites have been collected with the National Research Council (NRC) 1.8m Plaskett telescope located at the Dominion Astrophysical Observatory (DAO) in Victoria, Canada. The objective of this ongoing experiment is to study how reflectance spectroscopy can be used to reliably identify specific material types on the surface of artificial Earth-orbiting objects. Active geostationary satellites were selected as the main subjects for this experiment since their orientation is stable and can be estimated to a high-level of confidence throughout a night of observation. Furthermore, for most geostationary satellites, there is a wide variety of sources that can provide some level of information as to their external surface composition. Notwithstanding the high number of measurements that have been collected to date, it was assumed that the experimenters would have a much greater success rate in material identification given the choice experimental subjects. To date, only the presence of aluminum has been confidently identified in some of the reflectance spectra that have been collected. Two additional material types, namely photovoltaic cells and polyimide film, the first layer of multi-layer insulation (MLI), have also been possibly identified. However uncertainties in the reduced spectral measurements prevent any definitive conclusion with respect to these materials at this time. The surprising lack of results with respect to material identification have forced the experimenters to use other data interpretation methods to characterize the spectral scattering characteristics of the studied satellites. The results from this study have already led to improvements in the ways that reflectance spectra from spacecraft are collected and analysed. Equally important, the data interpretation techniques elaborated over the course of this experiment will also serve to increase the body of

  7. Automated measurement of site-specific N-glycosylation occupancy with SWATH-MS.

    PubMed

    Xu, Ying; Bailey, Ulla-Maja; Schulz, Benjamin L

    2015-07-01

    Asparagine-linked glycosylation is a common post-translational modification of proteins catalyzed by oligosaccharyltransferase that is important in regulating many aspects of protein function. Analysis of protein glycosylation, including glycoproteomic measurement of the site-specific extent of glycosylation, remains challenging. Here, we developed methods combining enzymatic deglycosylation and protease digestion with SWATH-MS to enable automated measurement of site-specific occupancy at many glycosylation sites. Deglycosylation with peptide-endoglycosidase H, leaving a remnant N-acetylglucosamine on asparagines previously carrying high-mannose glycans, followed by trypsin digestion allowed robust automated measurement of occupancy at many sites. Combining deglycosylation with the more general peptide-N-glycosidase F enzyme with AspN protease digest allowed robust automated differentiation of nonglycosylated and deglycosylated forms of a given glycosylation site. Ratiometric analysis of deglycosylated peptides and the total intensities of all peptides from the corresponding proteins allowed relative quantification of site-specific glycosylation occupancy between yeast strains with various isoforms of oligosaccharyltransferase. This approach also allowed robust measurement of glycosylation sites in human salivary glycoproteins. This method for automated relative quantification of site-specific glycosylation occupancy will be a useful tool for research with model systems and clinical samples. PMID:25737293

  8. Using Multiple Choice Examination Items To Measure Teachers' Content-Specific Pedagogical Knowledge.

    ERIC Educational Resources Information Center

    Kromrey, Jeffrey D.; Renfrow, Donata D.

    The use of multiple-choice test items measuring content-specific pedagogical knowledge (C-P) as a viable method of increasing the validity of teacher tests is described. The purposes of the paper are to: (1) present examples of multiple-choice test items used for the assessment of C-P and contrast these items with items used for assessing content…

  9. 12 CFR 3.210 - Standardized measurement method for specific risk

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... risk 3.210 Section 3.210 Banks and Banking COMPTROLLER OF THE CURRENCY, DEPARTMENT OF THE TREASURY CAPITAL ADEQUACY STANDARDS Risk-Weighted Assets-Market Risk § 3.210 Standardized measurement method for specific risk (a) General requirement. A national bank or Federal savings association must calculate...

  10. Initial Validation of an Instrument Measuring Psychology-Specific Epistemological Beliefs

    ERIC Educational Resources Information Center

    Renken, Maggie D.; McMahan, Ethan A.; Nitkova, Martina

    2015-01-01

    Psychology-specific epistemological beliefs (EBs) are believed to influence students' approach to and performance in psychology courses. However, empirical research on this topic is limited due in part to a lack of well-validated instruments measuring this construct. The primary objective of this research was to develop and validate the…

  11. Tense Marking and Spontaneous Speech Measures in Spanish Specific Language Impairment: A Discriminant Function Analysis

    ERIC Educational Resources Information Center

    Grinstead, John; Baron, Alisa; Vega-Mendoza, Mariana; De la Mora, Juliana; Cantu-Sanchez, Myriam; Flores, Blanca

    2013-01-01

    Purpose: To test the proposal that the tense deficit that has been demonstrated for children with specific language impairment (SLI) in other languages is also found in child Spanish and that low performance on tense-related measures can distinguish Spanish-speaking children with SLI from those without. Method: The authors evaluated evidence from…

  12. Measuring State-Specific Rumination: Development of the Rumination about an Interpersonal Offense Scale

    ERIC Educational Resources Information Center

    Wade, Nathaniel G.; Vogel, David L.; Liao, Kelly Yu-Hsin; Goldman, Daniel B.

    2008-01-01

    The tendency to ruminate has been consistently linked to psychological disturbances, such as increased stress, anger, and fear in response to provocations. However, existing measures of rumination focus on the disposition to ruminate rather than on rumination about a specific situation. This limits the ability to explore rumination about a…

  13. 78 FR 22883 - AHRQ Standing Workgroup for Quality Indicator Measure Specification

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-17

    ... HUMAN SERVICES Agency for Healthcare Research and Quality AHRQ Standing Workgroup for Quality Indicator Measure Specification AGENCY: Agency for Healthcare Research and Quality (AHRQ), HHS. ACTION: Notice of request for nominations. SUMMARY: The Agency for Healthcare Research and Quality (AHRQ) is...

  14. 78 FR 5810 - AHRQ Standing Workgroup for Quality Indicator Measure Specification

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-28

    ... HUMAN SERVICES Agency for Healthcare Research and Quality AHRQ Standing Workgroup for Quality Indicator Measure Specification AGENCY: Agency for Healthcare Research and Quality (AHRQ), HHS. ACTION: Notice of request for nominations. SUMMARY: The Agency for Healthcare Research and Quality (AHRQ) is...

  15. SITE-SPECIFIC PROTOCOL FOR MEASURING SOIL RADON POTENTIALS FOR FLORIDA HOUSES

    EPA Science Inventory

    The report describes a protocol for site-specific measurement of radon potentials for Florida houses that is consistent with existing residential radon protection maps. The protocol gives further guidance on the possible need for radon-protective house construction features. In a...

  16. Soft x-ray magneto-optic Kerr rotation and element-specific hysteresis measurement

    SciTech Connect

    Kortright, J.B.; Rice, M.

    1996-03-01

    Soft x-ray magneto-optic Kerr rotation has been measured using a continuously tunable multilayer linear polarizer in the beam reflected form samples in applied magnetic fields. Like magnetic circular dichroism, Kerr rotation in the soft x-ray can be element - specific and much larger than in the visible spectral range when the photon energy is tuned near atomic core resonances. Thus sensitive element-specific hysteresis measurements are possible with this technique. Examples showing large Kerr rotation from an Fe film and element-specific hysteresis loops of the Fe and Cr in an Fe/Cr multilayer demonstrate these new capabilities. Some consequences of the strong anomalous dispersion near the FeL{sub 2,3} edges to the Kerr rotation are discussed.

  17. Income and Physical Activity among Adults: Evidence from Self-Reported and Pedometer-Based Physical Activity Measurements

    PubMed Central

    Kari, Jaana T.; Pehkonen, Jaakko; Hirvensalo, Mirja; Yang, Xiaolin; Hutri-Kähönen, Nina; Raitakari, Olli T.; Tammelin, Tuija H.

    2015-01-01

    This study examined the relationship between income and physical activity by using three measures to illustrate daily physical activity: the self-reported physical activity index for leisure-time physical activity, pedometer-based total steps for overall daily physical activity, and pedometer-based aerobic steps that reflect continuous steps for more than 10 min at a time. The study population consisted of 753 adults from Finland (mean age 41.7 years; 64% women) who participated in 2011 in the follow-up of the ongoing Young Finns study. Ordinary least squares models were used to evaluate the associations between income and physical activity. The consistency of the results was explored by using register-based income information from Statistics Finland, employing the instrumental variable approach, and dividing the pedometer-based physical activity according to weekdays and weekend days. The results indicated that higher income was associated with higher self-reported physical activity for both genders. The results were robust to the inclusion of the control variables and the use of register-based income information. However, the pedometer-based results were gender-specific and depended on the measurement day (weekday vs. weekend day). In more detail, the association was positive for women and negative or non-existing for men. According to the measurement day, among women, income was positively associated with aerobic steps despite the measurement day and with totals steps measured on the weekend. Among men, income was negatively associated with aerobic steps measured on weekdays. The results indicate that there is an association between income and physical activity, but the association is gender-specific and depends on the measurement type of physical activity. PMID:26317865

  18. Carbon-14 Specific Activity Model Validation for Biota in Wetland Environments

    SciTech Connect

    Yankovich, T.L.; Sharp, K.J.; Benz, M.L.; Carr, J.; Killey, R.W.D.

    2008-01-15

    In many cases, contaminants, such as radionuclides, can show highly localized spatial distributions in natural systems. Therefore, a key question for environmental assessment and monitoring becomes, how can these localized distributions of contaminants in the environment lead to organism exposure, and ultimately, the potential for effects to receptor biota? To address this question, an important first step is to conduct field surveys at sites of interest to map out the spatial distribution and extent of contaminants in areas that are being occupied and utilized by resident receptor biota. Work can then be conducted to establish predictive relationships between contaminant concentrations in biota tissues and those in environmental media with which biota interact, to gain an understanding of how representative ambient contaminant concentrations are of biota exposure. The objectives of this study were: - To conduct a field survey in a wetland ecosystem to characterize the spatial distribution of carbon- 14 ({sup 14}C), a radionuclide with dynamics in natural systems that can be described using a specific activity model; and - To determine whether {sup 14}C concentrations in environmental media reflect those measured in tissues of resident flora and fauna. A detailed field campaign was carried out in summer 2001 to characterize the spatial distribution and areal coverage of {sup 14}C in Duke Swamp, a wetland ecosystem on Atomic Energy of Canada Limited (AECL)'s Chalk River Laboratories (CRL) site that receives {sup 14}C through releases from an up-gradient Waste Management Area (WMA), primarily through groundwater influx. Sampling of surface vegetation (dominantly comprised of Sphagnum moss) was conducted at a total of 69 locations, with complementary sampling of air, soil, fungi, aerial insects, ground-dwelling insects, amphibians, small mammals and snakes being carried out at a subset of five locations with varying {sup 14}C concentrations. Concentrations of {sup 14

  19. Highly specific and potently activating Vγ9Vδ2-T cell specific nanobodies for diagnostic and therapeutic applications.

    PubMed

    de Bruin, Renée C G; Lougheed, Sinéad M; van der Kruk, Liza; Stam, Anita G; Hooijberg, Erik; Roovers, Rob C; van Bergen En Henegouwen, Paul M P; Verheul, Henk M W; de Gruijl, Tanja D; van der Vliet, Hans J

    2016-08-01

    Vγ9Vδ2-T cells constitute the predominant subset of γδ-T cells in human peripheral blood and have been shown to play an important role in antimicrobial and antitumor immune responses. Several efforts have been initiated to exploit these cells for cancer immunotherapy, e.g. by using phosphoantigens, adoptive cell transfer, and by a bispecific monoclonal antibody based approach. Here, we report the generation of a novel set of Vγ9Vδ2-T cell specific VHH (or nanobody). VHH have several advantages compared to conventional antibodies related to their small size, stability, ease of generating multispecific molecules and low immunogenicity. With high specificity and affinity, the anti-Vγ9Vδ2-T cell receptor VHHs are shown to be useful for FACS, MACS and immunocytochemistry. In addition, some VHH were found to specifically activate Vγ9Vδ2-T cells. Besides being of possible immunotherapeutic value, these single domain antibodies will be of great value in the further study of this important immune effector cell subset. PMID:27373969

  20. Self-Efficacy and Social Support as Mediators Between Culturally Specific Dance and Lifestyle Physical Activity

    PubMed Central

    Murrock, Carolyn J.; Madigan, Elizabeth

    2013-01-01

    Culturally specific dance has the potential to generate health benefits but is seldom used even among studies advocating culturally specific interventions. This study examined the components of self-efficacy and social support as mediators between culturally specific dance and lifestyle physical activity in African American women (N = 126). An experimental design compared intervention and control groups for mediating effects of self-efficacy and social support on lifestyle physical activity. Findings indicated that only outcome expectations and social support from friends mediated effects. Culturally specific dance is a first step in encouraging African American women to become more physically active and improve health outcomes. The implications are that culturally specific dance programs can improve health outcomes by including members of underserved populations. PMID:18763475

  1. Quantifying Ant Activity Using Vibration Measurements

    PubMed Central

    Oberst, Sebastian; Baro, Enrique Nava; Lai, Joseph C. S.; Evans, Theodore A.

    2014-01-01

    Ant behaviour is of great interest due to their sociality. Ant behaviour is typically observed visually, however there are many circumstances where visual observation is not possible. It may be possible to assess ant behaviour using vibration signals produced by their physical movement. We demonstrate through a series of bioassays with different stimuli that the level of activity of meat ants (Iridomyrmex purpureus) can be quantified using vibrations, corresponding to observations with video. We found that ants exposed to physical shaking produced the highest average vibration amplitudes followed by ants with stones to drag, then ants with neighbours, illuminated ants and ants in darkness. In addition, we devised a novel method based on wavelet decomposition to separate the vibration signal owing to the initial ant behaviour from the substrate response, which will allow signals recorded from different substrates to be compared directly. Our results indicate the potential to use vibration signals to classify some ant behaviours in situations where visual observation could be difficult. PMID:24658467

  2. Infiltration rate measurement by active perfluorocarbon monitoring

    SciTech Connect

    Menzies, K.T.; Pong, C.M.; Randel, M.A. )

    1987-01-01

    The rate of air infiltration in homes and buildings is a significant factor affecting the magnitude of human exposure to air pollutants in the indoor environment. Several techniques have been utilized for the determination of air infiltration. These include building pressurization and tracer analysis, e.g., SF/sub 6/. Dietz and Cote at Brookhaven National Laboratory (BNL) have developed a simple, steady-state tracer kit that can be utilized by homeowners. This kit includes a source(s) of perfluorocarbon, i.e., perfluoromethylcyclohexane (PMCH) or perfluorodimethylcyclohexane (PDCH), and a passive sampling tube containing Ambersorb XE-347. Typically, the sampling tube is deployed for several days and then returned to a laboratory for analysis by thermal desorption/gas chromatography/electron capture detection. The authors developed an alternative sampling and analysis technique for PMCH/PDCH in homes. In order to facilitate monitoring of short-term infiltration rates (i.e., less than one day) they developed an active sorbent sampling method and solvent desorption/gas chromatography/electron capture detection analytical method. The method is based on the collection of PMCH on charcoal. The method validation, which is discussed in this article, includes analytical method development, selection of a solid sorbent, determination of desorption efficiency, analysis of breakthrough, testing of storage stability, and assessment of precision and accuracy in both the laboratory and field environment.

  3. Soft-x-ray magneto-optical Kerr effect and element-specific hysteresis measurement

    SciTech Connect

    Kortright, J.B.; Rice, M.

    1997-04-01

    Interest in the utilization of x-ray magneto-optical properties to provide element-specific magnetic information, combined with recent development of tunable linear polarizers for spectroscopic polarization measurement, have led the authors to the study of magneto-optical rotation (MOR) near core levels of magnetic atoms in magnetic multilayer and alloy films. Their initial observation of Faraday rotation (in transmission) demonstrated that for Fe MOR is easily measured and is larger at its L{sub 3} resonance than in the near-visible spectral regions. This work also demonstrated that the spectroscopic behavior of the MOR signal in transmission, resulting from the differential reaction of left- and right-circular components of a linearly polarized beam, is related to the magnetic circular dichroism (MCD), or differential absorption, as expected by a Kramers-Kronig transformation. Thus MCD measurements using circular polarization and MOR measurements using linear polarization can provide complementary, and in some cases equivalent, information. On beamline 6.3.2 the authors have begun to investigate soft x-ray MOR in the reflection geometry, the x-ray magneto-optic Kerr effect (XMOKE). Early measurements have demonstrated the ability to measure element-specific hysteresis loops and large rotations compared to analogous near-visible measurements. The authors are investigating the spectral dependence of the XMOKE signal, and have initiated systematic materials studies of sputter-deposited films of Fe, Fe{sub x}Cr{sub 1{minus}x} alloys, and Fe/Cr multilayers.

  4. The Measurement of Physical Activity in Young Children.

    ERIC Educational Resources Information Center

    Noland, Melody; And Others

    1990-01-01

    Presents data from two studies determining the validity of various measures of physical activity in preschool children. One explored how certain measures of activity predicted observed behavior. The other examined the Caltrac motion sensor's predictive validity. Results indicate the Caltrac monitor is sensitive to children's individual differences…

  5. 12 CFR 1806.201 - Measuring and reporting Qualified Activities.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 12 Banks and Banking 7 2010-01-01 2010-01-01 false Measuring and reporting Qualified Activities. 1806.201 Section 1806.201 Banks and Banking COMMUNITY DEVELOPMENT FINANCIAL INSTITUTIONS FUND, DEPARTMENT OF THE TREASURY BANK ENTERPRISE AWARD PROGRAM Awards § 1806.201 Measuring and reporting Qualified Activities. (a) General. An Applicant...

  6. Measuring Reading Activity: An Inventory. Instructional Resource No. 4.

    ERIC Educational Resources Information Center

    Guthrie, John T.; And Others

    Noting that the amount of reading students do is related to their reading achievement, this booklet presents an instrument designed to measure the amount and breadth of students' reading in and out of school. The first part of the booklet discusses the Reading Activity Inventory (RAI) and how it differs from other reading activity measures, uses…

  7. A Measurement Activity to Encourage Exploration of Calculus Concepts

    ERIC Educational Resources Information Center

    McGuffey, William

    2015-01-01

    This article describes an activity that incorporates measurement in order to lead students to discover and explore fundamental concepts of calculus. Students are provided with an experientially real starting point involving measurement and are encouraged to engage in mathematical discussions in a low-stakes environment. I describe the activity as…

  8. Twin Knudsen Cell Configuration for Activity Measurements by Mass Spectrometry

    NASA Technical Reports Server (NTRS)

    Jacobson, N. S.

    1996-01-01

    A twin Knudsen cell apparatus for alloy activity measurements by mass spectrometry is described. Two Knudsen cells - one containing an alloy and one containing a pure component - are mounted on a single flange and translated into the sampling region via a motorized x-y table. Mixing of the molecular beams from the cells is minimized by a novel system of shutters. Activity measurements were taken on two well-characterized alloys to verify the operation of the system. Silver activity measurements are reported for Ag-Cu alloys and aluminum activity measurements are reported for Fe-Al alloys. The temperature dependence of activity for a 0.474 mol fraction Al-Fe alloy gives a partial molar heat of aluminum. Measurements taken with the twin cell show good agreement with literature values for these alloys.

  9. The Role of Specificity, Targeted Learning Activities, and Prior Knowledge for the Effects of Relevance Instructions

    ERIC Educational Resources Information Center

    Roelle, Julian; Lehmkuhl, Nina; Beyer, Martin-Uwe; Berthold, Kirsten

    2015-01-01

    In 2 experiments we examined the role of (a) specificity, (b) the type of targeted learning activities, and (c) learners' prior knowledge for the effects of relevance instructions on learning from instructional explanations. In Experiment 1, we recruited novices regarding the topic of atomic structure (N = 80) and found that "specific"…

  10. Comprehensive measurement of respiratory activity in permeabilized cells using extracellular flux analysis

    PubMed Central

    Salabei, Joshua K.; Gibb, Andrew A.; Hill, Bradford G.

    2014-01-01

    Extracellular flux (XF) analysis has become a mainstream method to measure bioenergetic function in cells and tissues. While this technique is commonly used to measure energetics in intact cells, we outline here a detailed XF protocol for measuring respiration in permeabilized cells. Cells are permeabilized using saponin, digitonin, or recombinant perfringolysin O (XF PMP reagent) and provided with specific substrates to measure complex I- or II-mediated respiratory activity, Complex III+IV respiratory activity, or Complex IV activity. Medium- and long-chain acylcarnitines or glutamine may also be provided for measuring fatty acid oxidation or glutamine oxidation, respectively. This protocol allows for such measurements using a minimal number of cells compared with other protocols, without the need for mitochondrial isolation. The results are highly reproducible, and mitochondria remain well coupled. Collectively, this protocol provides comprehensive and detailed information regarding mitochondrial activity and efficiency, and, following preparative steps, takes approximately 6 hours to complete. PMID:24457333

  11. Measurement of uterine activity in vitro by integrating muscle tension

    PubMed Central

    Styles, P. R.; Sullivan, T. J.

    1962-01-01

    Spontaneous or electrically stimulated activity of the uterus is measured isometrically in vitro by integrating tension against time. Uterine contractions move the operating rod of a potentiometer transducer, the output voltage from which is coupled to an electrical integrator motor and a servo recorder. Several parameters of uterine activity can be expressed in a single measurement, and a record of isometric contractions is obtained simultaneously. Oxytocin can be assayed accurately and the effect of drugs on uterine motility can be measured. PMID:13918066

  12. Active Learning: The Importance of Developing a Comprehensive Measure

    ERIC Educational Resources Information Center

    Carr, Rodney; Palmer, Stuart; Hagel, Pauline

    2015-01-01

    This article reports on an investigation into the validity of a widely used scale for measuring the extent to which higher education students employ active learning strategies. The scale is the active learning scale in the Australasian Survey of Student Engagement. This scale is based on the Active and Collaborative Learning scale of the National…

  13. Cancer-specific binary expression system activated in mice by bacteriophage HK022 Integrase.

    PubMed

    Elias, Amer; Spector, Itay; Sogolovsky-Bard, Ilana; Gritsenko, Natalia; Rask, Lene; Mainbakh, Yuli; Zilberstein, Yael; Yagil, Ezra; Kolot, Mikhail

    2016-01-01

    Binary systems based on site-specific recombination have been used for tumor specific transcription targeting of suicide genes in animal models. In these binary systems a site specific recombinase or integrase that is expressed from a tumor specific promoter drives tumor specific expression of a cytotoxic gene. In the present study we developed a new cancer specific binary expression system activated by the Integrase (Int) of the lambdoid phage HK022. We demonstrate the validity of this system by the specific expression of a luciferase (luc) reporter in human embryonic kidney 293T (HEK293T) cells and in a lung cancer mouse model. Due to the absence viral vectors and of cytotoxicity the Int based binary system offers advantages over previously described counterparts and may therefore be developed into a safer cancer cell killing system. PMID:27117628

  14. Cancer-specific binary expression system activated in mice by bacteriophage HK022 Integrase

    PubMed Central

    Elias, Amer; Spector, Itay; Sogolovsky-Bard, Ilana; Gritsenko, Natalia; Rask, Lene; Mainbakh, Yuli; Zilberstein, Yael; Yagil, Ezra; Kolot, Mikhail

    2016-01-01

    Binary systems based on site-specific recombination have been used for tumor specific transcription targeting of suicide genes in animal models. In these binary systems a site specific recombinase or integrase that is expressed from a tumor specific promoter drives tumor specific expression of a cytotoxic gene. In the present study we developed a new cancer specific binary expression system activated by the Integrase (Int) of the lambdoid phage HK022. We demonstrate the validity of this system by the specific expression of a luciferase (luc) reporter in human embryonic kidney 293T (HEK293T) cells and in a lung cancer mouse model. Due to the absence viral vectors and of cytotoxicity the Int based binary system offers advantages over previously described counterparts and may therefore be developed into a safer cancer cell killing system. PMID:27117628

  15. Prostate-specific antigen as a marker of disease activity in prostate cancer.

    PubMed

    Partin, Alan W; Hanks, Gerald E; Klein, Eric A; Moul, Judd W; Nelson, William G; Scher, Howard I

    2002-09-01

    Despite the impact of prostate-specific antigen (PSA) testing on the detection and management of prostate cancer, controversy about its usefulness as a marker of disease activity continues. This review, based on a recent roundtable discussion, examines whether PSA measurements can be used rationally in several clinical settings. Following radical prostatectomy and radiation therapy, prediction of survival by PSA level is most reliable in high-risk patients. PSA doubling time after radiation therapy is the strongest predictor of biochemical failure. PSA measurements have been associated with inconsistent results following hormonal treatment; reduced PSA levels may result from antiandrogen treatment, which decreases expression of the PSA gene, and therefore, the level of PSA production. In the setting of primary and secondary cancer prevention, PSA is important in risk stratification when selecting patients for studies. Part 2 of this two-part article, which began in the August issue, discusses the role of PSA in hormonal and drug therapies and in primary and secondary chemoprevention. PMID:12380948

  16. Lentivirally Engineered DC activate AFP-specific T cells which Inhibit Hepatocellular Carcinoma Growth in vitro and in vivo

    PubMed Central

    Liu, Yang; Butterfield, Lisa H.; Fu, Xiaohui; Song, Zhenshun; Zhang, Xiaoping; Lu, Chongde; Ding, Guanghui; Wu, Mengchao

    2012-01-01

    Alpha-fetoprotein (AFP), a tumor-associated antigen for hepatocellular carcinoma (HCC), is an established biomarker for HCC. In this study, we created a lentivirus expressing the AFP antigen and investigated the antitumor activity of AFP-specific CD8+ T cells, with and without CD4+ T cells, which were activated by either AFP peptide-pulsed or Lenti-AFP-engineered DC in vitro and in vivo. AFP-specific T cells could efficiently kill HepG2 HCC cells, and produced IL-2, IFN-γ, TNF-α, perforin and granzyme B, with minimal production of IL-10 (a negative regulator of T cell activation). Both strategies activated AFP-specific T cells, but the lentiviral strategy was superior by several measures. Data also support an impact of CD4+ T cells in supporting anti-tumor activity. In vivo studies in a xenograft HCC tumor model also showed that AFP-specific T cells could markedly suppress HCC tumor formation and morbidity in tumor-bearing nude mice, as well as regulate serum levels of related cytokines and antitumor molecules. In parallel with human in vitro T cell cultures, the in vivo model demonstrated superior anti-tumor effects and Th1-skewing with Lenti-AFP-DC. This study supports the superiority of a full-length antigen lentivirus-based DC vaccine strategy over peptides, and provides new insight into the design of DC-based vaccines. PMID:21491085

  17. Resin {sup 90}Y microsphere activity measurements for liver brachytherapy

    SciTech Connect

    Dezarn, William A.; Kennedy, Andrew S.

    2007-06-15

    The measurement of the radioactivity administered to the patient is one of the major components of {sup 90}Y microsphere liver brachytherapy. The activity of {sup 90}Y microspheres in a glass delivery vial was measured in a dose calibrator. The calibration value to use for {sup 90}Y in the dose calibrator was verified using an activity calibration standard provided by the microsphere manufacturer. This method allowed for the determination of a consistent, reproducible local activity standard. Additional measurements were made to determine some of the factors that could affect activity measurement. The axial response of the dose calibrator was determined by the ratio of activity measurements at the bottom and center of the dose calibrator. The axial response was 0.964 for a glass shipping vial, 1.001 for a glass V-vial, and 0.988 for a polycarbonate V-vial. Comparisons between activity measurements in the dose calibrator and those using a radiation survey meter were found to agree within 10%. It was determined that the dose calibrator method was superior to the survey meter method because the former allowed better defined measurement geometry and traceability of the activity standard back to the manufacturer. Part of the preparation of resin {sup 90}Y microspheres for patient delivery is to draw out a predetermined activity from a shipping vial and place it into a V-vial for delivery to the patient. If the drawn activity was placed in a glass V-vial, the activity measured in the dose calibrator with a glass V-vial was 4% higher than the drawn activity from the shipping vial standard. If the drawn activity was placed in a polycarbonate V-vial, the activity measured in the dose calibrator with a polycarbonate V-vial activity was 20% higher than the drawn activity from the shipping vial standard. Careful characterization of the local activity measurement standard is recommended instead of simply accepting the calibration value of the dose calibrator manufacturer.

  18. Bacteriophage PSP3 and phiR73 activator proteins: analysis of promoter specificities.

    PubMed Central

    Julien, B; Calendar, R

    1996-01-01

    Transcription from the late promoters of bacteriophage P2 and its satellite phage P4 is activated by a unique class of small, zinc-binding proteins. Using plasmid expression systems, we compared activators from two P2-like (helper) phages with those encoded by two satellite phages. The helper phage activators have more activity on the P4 phage sid promoter. In contrast, the satellite phage activators function better on the four late P2 promoters and on the P4 late leftward promoter. We purified one activator encoded by a P2-like phage and an activator from a satellite phage and determined their binding sites within the P2 and P4 late promoters. Differences in activity levels correlate with binding specificities; promoters that function best with the satellite phage activators have only one activator binding site centered at -55, while the P4 sid promoter, which has more activity with helper phage activators, has a second binding site centered at -18. Surprisingly, DNase I footprinting revealed only very minor differences in promoter binding by the two activators reported here and the P4 activator reported previously. Thus, the differences in transcriptional activity are probably due to interactions between the activators and RNA polymerase, rather than interactions between the activators and DNA. PMID:8824611

  19. Specific activities of antioxidative enzymes in the cochlea of guinea pigs at different stages of development.

    PubMed

    Zelck, U; Nowak, R; Karnstedt, U; Koitschev, A; Käcker, N

    1993-01-01

    Significant activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were found in the cochleas of guinea pigs of different ages. The specific activities of SOD and GSH-Px (expressed as units/mg protein) increased significantly from fetal animals to animals 2 days old and then to 6-month-old animals. PMID:8369116

  20. Activation of enhancer elements by the homeobox gene Cdx2 is cell line specific.

    PubMed Central

    Taylor, J K; Levy, T; Suh, E R; Traber, P G

    1997-01-01

    Cdx2 is a caudal-related homeodomain transcription factor that is expressed in complex patterns during mouse development and at high levels in the intestinal epithelium of adult mice. Cdx2 activates transcription of intestinal gene promoters containing specific binding sites. Moreover, Cdx2 has been shown to induce intestinal differentiation in cell lines. In this study, we show that Cdx2 is able to bind to two well defined enhancer elements in the HoxC8 gene. We then demonstrate that Cdx2 is able to activate transcription of heterologous promoters when its DNA binding element is placed in an enhancer context. Furthermore, the ability to activate enhancer elements is cell-line dependent. When the Cdx2 activation domain was linked to the Gal4 DNA binding domain, the chimeric protein was able to activate Gal4 enhancer constructs in an intestinal cell line, but was unable to activate transcription in NIH3T3 cells. These data suggest that there are cell-specific factors that allow the Cdx2 activation domain to function in the activation of enhancer elements. We hypothesize that either a co-activator protein or differential phosphorylation of the activation domain may be the mechanism for intestinal cell line-specific function of Cdx2 and possibly in other tissues in early development. PMID:9171078

  1. Charged fusion product loss measurements using nuclear activation.

    PubMed

    Bonheure, G; Hult, M; González de Orduña, R; Arnold, D; Dombrowski, H; Laubenstein, M; Wieslander, E; Vermaercke, P; Murari, A; Popovichev, S; Mlynar, J

    2010-10-01

    In ITER, α particle loss measurements will be required in order to understand the alpha particle physics. Techniques capable of operating in a fusion reactor environment need further development. Recent experimental studies on JET demonstrated the potential of nuclear activation to measure the flux of escaping MeV ions. New results from MeV ion induced activation of metallic, ceramic, and crystal samples placed near the plasma edge are reported. Activation products were measured as function of orientation with respect to the magnetic field as well as function of the distance to the plasma. Sample activity was measured using ultralow-level gamma-ray spectrometry. Distribution of 14.68 MeV fusion proton induced activation products is strongly anisotropic in agreement with simulations and falls off sharply with increasing distance to the plasma. Prospects for using the technique in ITER are discussed. PMID:21058458

  2. Threshold occupancy and specific cation binding modes in the hammerhead ribozyme active site are required for active conformation

    PubMed Central

    Lee, Tai-Sung; Giambaşu, George M.; Sosa, Carlos P.; Martick, Monika; Scott, William G.; York, Darrin M.

    2009-01-01

    The relationship between formation of active in-line attack conformations and monovalent (Na+) and divalent (Mg2+) metal ion binding in the hammerhead ribozyme has been explored with molecular dynamics simulations. To stabilize repulsions between negatively charged groups, different requirements of threshold occupancy of metal ions were observed in the reactant and activated precursor states both in the presence or absence of a Mg2+ in the active site. Specific bridging coordination patterns of the ions are correlated with the formation of active in-line attack conformations and can be accommodated in both cases. Furthermore, simulation results suggest that the hammerhead ribozyme folds to form an electronegative recruiting pocket that attracts high local concentrations of positive charge. The present simulations help to reconcile experiments that probe the metal ion sensitivity of hammerhead ribozyme catalysis and support the supposition that Mg2+, in addition to stabilizing active conformations, plays a specific chemical role in catalysis. PMID:19265710

  3. Accuracy, Precision, Sensitivity, and Specificity of Noninvasive ICP Absolute Value Measurements.

    PubMed

    Krakauskaite, Solventa; Petkus, Vytautas; Bartusis, Laimonas; Zakelis, Rolandas; Chomskis, Romanas; Preiksaitis, Aidanas; Ragauskas, Arminas; Matijosaitis, Vaidas; Petrikonis, Kestutis; Rastenyte, Daiva

    2016-01-01

    An innovative absolute intracranial pressure (ICP) value measurement method has been validated by multicenter comparative clinical studies. The method is based on two-depth transcranial Doppler (TCD) technology and uses intracranial and extracranial segments of the ophthalmic artery as pressure sensors. The ophthalmic artery is used as a natural pair of "scales" that compares ICP with controlled pressure Pe, which is externally applied to the orbit. To balance the scales, ICP = Pe a special two-depth TCD device was used as a pressure balance indicator. The proposed method is the only noninvasive ICP measurement method that does not need patient-specific calibration. PMID:27165929

  4. Specific heat measurement set-up for quench condensed thin superconducting films.

    PubMed

    Poran, Shachaf; Molina-Ruiz, Manel; Gérardin, Anne; Frydman, Aviad; Bourgeois, Olivier

    2014-05-01

    We present a set-up designed for the measurement of specific heat of very thin or ultra-thin quench condensed superconducting films. In an ultra-high vacuum chamber, materials of interest can be thermally evaporated directly on a silicon membrane regulated in temperature from 1.4 K to 10 K. On this membrane, a heater and a thermometer are lithographically fabricated, allowing the measurement of heat capacity of the quench condensed layers. This apparatus permits the simultaneous thermal and electrical characterization of successively deposited layers in situ without exposing the deposited materials to room temperature or atmospheric conditions, both being irreversibly harmful to the samples. This system can be used to study specific heat signatures of phase transitions through the superconductor to insulator transition of quench condensed films. PMID:24880383

  5. Aluminum phosphide poisoning: Possible role of supportive measures in the absence of specific antidote

    PubMed Central

    Agrawal, Vijay Kumar; Bansal, Abhishek; Singh, Ranjeet Kumar; Kumawat, Bhanwar Lal; Mahajan, Parul

    2015-01-01

    Aluminum phosphide (ALP) poisoning is one of the major causes of suicidal deaths. Toxicity by ALP is caused by the liberation of phosphine gas, which rapidly causes cell hypoxia due to inhibition of oxidative phosphorylation, leading to circulatory failure. Treatment of ALP toxicity is mainly supportive as there is no specific antidote. We recently managed 7 cases of ALP poisoning with severe hemodynamic effects. Patients were treated with supportive measures including gastric lavage with diluted potassium permanganate, coconut oil and sodium-bicarbonate first person account should be avoided in a scientific paper. Intravenous magnesium sulfate, proper hemodynamic monitoring and vasopressors. Four out of 7 survived thus suggesting a role of such supportive measures in the absence of specific antidote for ALP poisoning. PMID:25722553

  6. Aluminum phosphide poisoning: Possible role of supportive measures in the absence of specific antidote.

    PubMed

    Agrawal, Vijay Kumar; Bansal, Abhishek; Singh, Ranjeet Kumar; Kumawat, Bhanwar Lal; Mahajan, Parul

    2015-02-01

    Aluminum phosphide (ALP) poisoning is one of the major causes of suicidal deaths. Toxicity by ALP is caused by the liberation of phosphine gas, which rapidly causes cell hypoxia due to inhibition of oxidative phosphorylation, leading to circulatory failure. Treatment of ALP toxicity is mainly supportive as there is no specific antidote. We recently managed 7 cases of ALP poisoning with severe hemodynamic effects. Patients were treated with supportive measures including gastric lavage with diluted potassium permanganate, coconut oil and sodium-bicarbonate first person account should be avoided in a scientific paper. Intravenous magnesium sulfate, proper hemodynamic monitoring and vasopressors. Four out of 7 survived thus suggesting a role of such supportive measures in the absence of specific antidote for ALP poisoning. PMID:25722553

  7. The core-specific lysosomal alpha(1-6)-mannosidase activity depends on aspartamidohydrolase activity.

    PubMed Central

    Haeuw, J F; Grard, T; Alonso, C; Strecker, G; Michalski, J C

    1994-01-01

    The substrate specificity of the core-specific rat liver lysosomal alpha(1-6)-mannosidase was investigated using mannosylated oligosaccharides and glycoasparagines. Hydrolysis of Man(alpha 1-6) linkage hydrolysis was demonstrated to follow the action of endoglycosidases, namely aspartyl-N-acetyl-beta-D-glucosaminidase and endo-N-acetyl-beta-D-glucosaminidase. The results are discussed with respect to the nature of the carbohydrate materials stored in the tissues and excreted in the urine from patients suffering from aspartylglucosaminuria and fucosidosis. Images Figure 1 Figure 2 PMID:8110182

  8. Method for measuring surface activity of silicon nitride powder

    NASA Technical Reports Server (NTRS)

    Kanno, Y.; Imai, H.

    1985-01-01

    Amorphous, alpha-, and beta-Si3N4 powders were activated by vibration ball milling in purified MeOH, and the surface activity of ground powders was determined by the temperature programmed desorption (TPD) method using NH3 gas. The concentration of active sites with a potential energy equivalent to the peak temperature in the spectrum increased was markedly by ball milling the amorphous Si3N4. The alpha- and beta-Si3N4 also had active sites produced by ball milling. The concentration of active site increased with increased ball milling time. A method for measuring surface activity of ceramic raw materials by TPD is proposed.

  9. Development of Portable Beta Spectrometer for Sr-90 Activity Measurements in Field Conditions and Its Application in Rehabilitation Activities at RRC Kurchatov Institute

    SciTech Connect

    Potapov, V.N.; Volkovich, A.G.; Ivanov, O.P.; Stepanov, V.E.; Smirnov, S.V.; Volkov, V.G.

    2006-07-01

    A new method to measure the Sr-90 ground specific activity in situ was developed. It is based on the count-rates determination in selected energy ranges of two registered apparatus spectra: total {beta} + {gamma} spectrum and {gamma} spectrum. A numerical simulation of the detector performance defined these energy ranges used for calculation of activity. For implementation of the proposed method a portable instrument was developed and manufactured. Parameters of the instrument are the following: the range of measurement for a specific activity mode - (60 - 3.0x10{sup 6}) Bq/kg; the range for total activity countable mode (0.5 - 2.0x10{sup 4}) Bq; minimum measurable specific activity Sr-90 for samples containing natural radionuclides - 60 Bq/kg, minimum measurable activity for samples not containing NRN - 0.5 Bq. (authors)

  10. Anti-human cytomegalovirus activity of cytokines produced by CD4+ T-cell clones specifically activated by IE1 peptides in vitro.

    PubMed Central

    Davignon, J L; Castanié, P; Yorke, J A; Gautier, N; Clément, D; Davrinche, C

    1996-01-01

    The control of latent cytomegalovirus (CMV) infections by the immune system is poorly understood. We have previously shown that CD4+ T cells specific for the human CMV major regulatory protein IE1 are frequent in latently infected healthy blood donors. In order to learn about the possible role of these cells, we have developed IE1-specific CD4+ T-cell clones and, in this study, analyzed their epitope specificity and function in vitro. We measured their cytokine production when stimulated with specific IE1 peptides or whole recombinant IE1 protein. Their cytokine profiles, as deduced from gamma interferon (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), and interleukin-4 (IL-4) and IL-6 production, were of the Th0- and Th1-like phenotypes. Supernatants from IE1-specific clones producing IFN-gamma and TNF-alpha were shown to inhibit CMV replication in U373 MG cells. This effect was due, as found by using cytokine-specific neutralizing antibodies, mostly to IFN-gamma, which was secreted at higher levels than TNF-alpha. To better assess the anti-CMV activity of cytokines, recombinant IFN-gamma and TNF-alpha were used and shown to have a synergistic effect on the inhibition of CMV replication and protein expression. Thus, IE1-specific CD4+ T cells display in vitro anti-CMV activity through cytokine secretion and may play a role in the control of in vivo latent infections. PMID:8642638

  11. Specific cognitive functions and depressive symptoms as predictors of activities of daily living in older adults with heterogeneous cognitive backgrounds

    PubMed Central

    de Paula, Jonas J.; Diniz, Breno S.; Bicalho, Maria A.; Albuquerque, Maicon Rodrigues; Nicolato, Rodrigo; de Moraes, Edgar N.; Romano-Silva, Marco A.; Malloy-Diniz, Leandro F.

    2015-01-01

    Cognitive functioning influences activities of daily living (ADL). However, studies reporting the association between ADL and neuropsychological performance show inconsistent results regarding what specific cognitive domains are related to each specific functional domains. Additionally, whether depressive symptoms are associated with a worse functional performance in older adults is still under explored. We investigated if specific cognitive domains and depressive symptoms would affect different aspects of ADL. Participants were 274 older adults (96 normal aging participants, 85 patients with mild cognitive impairment, and 93 patients probable with mild Alzheimer’s disease dementia) with low formal education (∼4 years). Measures of ADL included three complexity levels: Self-care, Instrumental-Domestic, and Instrumental-Complex. The specific cognitive functions were evaluated through a factorial strategy resulting in four cognitive domains: Executive Functions, Language/Semantic Memory, Episodic Memory, and Visuospatial Abilities. The Geriatric Depression Scale measured depressive symptoms. Multiple linear regression analysis showed executive functions and episodic memory as significant predictors of Instrumental-Domestic ADL, and executive functions, episodic memory and language/semantic memory as predictors of Instrumental-Complex ADL (22 and 28% of explained variance, respectively). Ordinal regression analysis showed the influence of specific cognitive functions and depressive symptoms on each one of the instrumental ADL. We observed a heterogeneous pattern of association with explained variance ranging from 22 to 38%. Different instrumental ADL had specific cognitive predictors and depressive symptoms were predictive of ADL involving social contact. Our results suggest a specific pattern of influence depending on the specific instrumental daily living activity. PMID:26257644

  12. Ice nucleus activity measurements of solid rocket motor exhaust particles

    NASA Technical Reports Server (NTRS)

    Keller, V. W. (Compiler)

    1986-01-01

    The ice Nucleus activity of exhaust particles generated from combustion of Space Shuttle propellant in small rocket motors has been measured. The activity at -20 C was substantially lower than that of aerosols generated by unpressurized combustion of propellant samples in previous studies. The activity decays rapidly with time and is decreased further in the presence of moist air. These tests corroborate the low effectivity ice nucleus measurement results obtained in the exhaust ground cloud of the Space Shuttle. Such low ice nucleus activity implies that Space Shuttle induced inadvertent weather modification via an ice phase process is extremely unlikely.

  13. Validation of subject-specific cardiovascular system models from porcine measurements.

    PubMed

    Revie, James A; Stevenson, David J; Chase, J Geoffrey; Hann, Christopher E; Lambermont, Bernard C; Ghuysen, Alexandre; Kolh, Philippe; Shaw, Geoffrey M; Heldmann, Stefan; Desaive, Thomas

    2013-02-01

    A previously validated mathematical model of the cardiovascular system (CVS) is made subject-specific using an iterative, proportional gain-based identification method. Prior works utilised a complete set of experimentally measured data that is not clinically typical or applicable. In this paper, parameters are identified using proportional gain-based control and a minimal, clinically available set of measurements. The new method makes use of several intermediary steps through identification of smaller compartmental models of CVS to reduce the number of parameters identified simultaneously and increase the convergence stability of the method. This new, clinically relevant, minimal measurement approach is validated using a porcine model of acute pulmonary embolism (APE). Trials were performed on five pigs, each inserted with three autologous blood clots of decreasing size over a period of four to five hours. All experiments were reviewed and approved by the Ethics Committee of the Medical Faculty at the University of Liege, Belgium. Continuous aortic and pulmonary artery pressures (P(ao), P(pa)) were measured along with left and right ventricle pressure and volume waveforms. Subject-specific CVS models were identified from global end diastolic volume (GEDV), stroke volume (SV), P(ao), and P(pa) measurements, with the mean volumes and maximum pressures of the left and right ventricles used to verify the accuracy of the fitted models. The inputs (GEDV, SV, P(ao), P(pa)) used in the identification process were matched by the CVS model to errors <0.5%. Prediction of the mean ventricular volumes and maximum ventricular pressures not used to fit the model compared experimental measurements to median absolute errors of 4.3% and 4.4%, which are equivalent to the measurement errors of currently used monitoring devices in the ICU (∼5-10%). These results validate the potential for implementing this approach in the intensive care unit. PMID:22126892

  14. Accuracy of Computational Cerebral Aneurysm Hemodynamics Using Patient-Specific Endovascular Measurements

    NASA Astrophysics Data System (ADS)

    McGah, Patrick; Levitt, Michael; Barbour, Michael; Mourad, Pierre; Kim, Louis; Aliseda, Alberto

    2013-11-01

    We study the hemodynamic conditions in patients with cerebral aneurysms through endovascular measurements and computational fluid dynamics. Ten unruptured cerebral aneurysms were clinically assessed by three dimensional rotational angiography and an endovascular guidewire with dual Doppler ultrasound transducer and piezoresistive pressure sensor at multiple peri-aneurysmal locations. These measurements are used to define boundary conditions for flow simulations at and near the aneurysms. The additional in vivo measurements, which were not prescribed in the simulation, are used to assess the accuracy of the simulated flow velocity and pressure. We also performed simulations with stereotypical literature-derived boundary conditions. Simulated velocities using patient-specific boundary conditions showed good agreement with the guidewire measurements, with no systematic bias and a random scatter of about 25%. Simulated velocities using the literature-derived values showed a systematic over-prediction in velocity by 30% with a random scatter of about 40%. Computational hemodynamics using endovascularly-derived patient-specific boundary conditions have the potential to improve treatment predictions as they provide more accurate and precise results of the aneurysmal hemodynamics. Supported by an R03 grant from NIH/NINDS

  15. Objectively measured physical activity of young Canadian children using accelerometry.

    PubMed

    Borkhoff, Cornelia M; Heale, Liane D; Anderson, Laura N; Tremblay, Mark S; Maguire, Jonathon L; Parkin, Patricia C; Birken, Catherine S

    2015-12-01

    The objective of the study was to describe objectively measured physical activity (PA) and sedentary time of infants, toddlers, and preschoolers and determine the proportion meeting Canadian age-specific PA guidelines. Ninety children (47 girls, 43 boys; mean age 32 (range, 4-70) months) attending scheduled health supervision visits and in the TARGet Kids! (The Applied Research Group for Kids) cohort wore an Actical accelerometer for 7 days. Participants with 4 or more valid days were included in the analysis. Time, in mean minutes per day (min/day), spent sedentary and in light PA, moderate to vigorous PA (MVPA), and total PA was determined using published cut-points; age groups were compared using ANOVA. Twenty-three percent of children <18 months (n = 28) and 76% of children aged 18-59 months (n = 45) met the guideline of 180 min/day of total PA; 13% of children ≥60 months (n = 17) met the guideline of 60 min/day of MVPA. Children <18 months spent more of their waking time per day engaged in sedentary behaviours (79%; ∼7.3 h) compared with children aged 18-59 months (63%; ∼6.6 h) and children ≥60 months (58%; ∼6.6 h). In conclusion, most children aged 18-59 months met the Canadian PA guidelines for children aged 0-4 years, whereas few younger than 18 months met the same guidelines. Only 13% of children ≥5 years met their age-specific PA guidelines. Further research is needed to develop, test, and implement effective strategies to promote PA and reduce sedentary behaviour in very young children. PMID:26573442

  16. Colourimetric and fluorometric substrates for measurement of pullulanase activity.

    PubMed

    McCleary, Barry V; Mangan, David; McKie, Vincent; Cornaggia, Claudio; Ivory, Ruth; Rooney, Edward

    2014-07-01

    Specific and highly sensitive colourimetric and fluorometric substrate mixtures have been prepared for the measurement of pullulanase and limit-dextrinase activity and assays employing these substrates have been developed. These mixtures comprise thermostable α- and β-glucosidases and either 4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-maltotriosyl (1-6) α-maltotrioside (BzCNPG3G3, 1) as a colourimetric substrate or 4,6-O-benzylidene-4-methylumbelliferyl-β-maltotriosyl (1-6) α-maltotrioside (BzMUG3G3, 2) as a fluorometric substrate. Hydrolysis of substrates 1 and 2 by exo-acting enzymes such as amyloglucosidase, β-amylase and α-glucosidase is prevented by the presence of the 4,6-O-benzylidene group on the non-reducing end D-glucosyl residue. The substrates are not hydrolysed by any α-amylases studied, (including those from Aspergillus niger and porcine pancreas) and are resistant to hydrolysis by Pseudomonas sp. isoamylase. On hydrolysis by pullulanase, the 2-chloro-4-nitrophenyl-β-maltotrioside (3) or 4-methylumbelliferyl-β-maltotrioside (4) liberated is immediately hydrolysed to D-glucose and 2-chloro-4-nitrophenol or 4-methylumbelliferone. The reaction is terminated by the addition of a weak alkaline solution leading to the formation of phenolate ions in solution whose concentration can be determined using either spectrophotometric or fluorometric analysis. The assay procedure is simple to use, specific, accurate, robust and readily adapted to automation. PMID:24938640

  17. Specific Activation of Estrogen Receptor Alpha and Beta Enhances Male Sexual Behavior and Neuroplasticity in Male Japanese Quail

    PubMed Central

    Seredynski, Aurore L.; Ball, Gregory F.; Balthazart, Jacques; Charlier, Thierry D.

    2011-01-01

    Two subtypes of estrogen receptors (ER), ERα and ERβ, have been identified in humans and numerous vertebrates, including the Japanese quail. We investigated in this species the specific role(s) of each receptor in the activation of male sexual behavior and the underlying estrogen-dependent neural plasticity. Castrated male Japanese quail received empty (CX) or testosterone-filled (T) implants or were daily injected with the ER general agonist diethylstilbestrol (DES), the ERα-specific agonist PPT, the ERβ-specific agonist DPN or the vehicle, propylene glycol. Three days after receiving the first treatment, subjects were alternatively tested for appetitive (rhythmic cloacal sphincter movements, RCSM) and consummatory aspects (copulatory behavior) of male sexual behavior. 24 hours after the last behavioral testing, brains were collected and analyzed for aromatase expression and vasotocinergic innervation in the medial preoptic nucleus. The expression of RCSM was activated by T and to a lesser extent by DES and PPT but not by the ERβagonist DPN. In parallel, T fully restored the complete sequence of copulation, DES was partially active and the specific activation of ERα or ERβ only resulted in a very low frequency of mount attempts in few subjects. T increased the volume of the medial preoptic nucleus as measured by the dense cluster of aromatase-immunoreactive cells and the density of the vasotocinergic innervation within this nucleus. DES had only a weak action on vasotocinergic fibers and the two specific ER agonists did not affect these neural responses. Simultaneous activation of both receptors or treatments with higher doses may be required to fully activate sexual behavior and the associated neurochemical events. PMID:21533185

  18. Preparation of activated carbon with large specific surface area from reed black liquor.

    PubMed

    Sun, Y; Zhang, J P; Yang, G; Li, Z H

    2007-05-01

    Activated carbon with large specific surface area and well-developed porosity was prepared from pyrolysis of K2CO3-impregnated lignin precipitated from reed pulp black liquors. The impregnation ratio was 1:1. The effect of activation temperature upon the Brunauer-Emmett-Teller (BET) specific surface area and pore volume of the carbon was closely investigated. Increasing activation temperature led to an opening and widening of the porous structure below 800'C. Above 800'C, the excess widening of pore led to the decrease of BET surface area and micropore volume. The BET surface area and pore volume of the carbon activated at 800 degrees C were 1395 m(2) g(-1) and 0.7702 ml g(-1) , respectively. The potential application of the carbon activated at 800 degrees C for removal of Cr (VI) was also investigated. The experimental results showed that it had good adsorption capacity. PMID:17615958

  19. Quasi-static magnetic measurements to predict specific absorption rates in magnetic fluid hyperthermia experiments

    NASA Astrophysics Data System (ADS)

    Coral, D. F.; Mendoza Zélis, P.; de Sousa, M. E.; Muraca, D.; Lassalle, V.; Nicolás, P.; Ferreira, M. L.; Fernández van Raap, M. B.

    2014-01-01

    In this work, the issue on whether dynamic magnetic properties of polydispersed magnetic colloids modeled using physical magnitudes derived from quasi-static magnetic measurement can be extrapolated to analyze specific absorption rate data acquired at high amplitudes and frequencies of excitation fields is addressed. To this end, we have analyzed two colloids of magnetite nanoparticles coated with oleic acid and chitosan in water displaying, under a radiofrequency field, high and low specific heat power release. Both colloids are alike in terms of liquid carrier, surfactant and magnetic phase composition but differ on the nanoparticle structuring. The colloid displaying low specific dissipation consists of spaced magnetic nanoparticles of mean size around 4.8 nm inside a large chitosan particle of 52.5 nm. The one displaying high specific dissipation consists of clusters of magnetic nanoparticles of mean size around 9.7 nm inside a chitosan particle of 48.6 nm. The experimental evaluation of Néel and Brown relaxation times (˜10-10 s and 10-4 s, respectively) indicate that the nanoparticles in both colloids magnetically relax by Néel mechanism. The isothermal magnetization curves analysis for this mechanism show that the magnetic nanoparticles behave in the interacting superparamagnetic regime. The specific absorption rates were determined calorimetrically at 260 kHz and up to 52 kA/m and were well modeled within linear response theory using the anisotropy density energy retrieved from quasi-static magnetic measurement, validating their use to predict heating ability of a given polydispersed particle suspension. Our findings provide new insight in the validity of quasi-static magnetic characterization to analyze the high frequency behavior of polydispersed colloids within the framework of the linear response and Wohlfarth theories and indicate that dipolar interactions play a key role being their strength larger for the colloid displaying higher dissipation, i

  20. Validity of critical frequency test for measuring table tennis aerobic endurance through specific protocol

    PubMed Central

    Zagatto, Alessandro M.; Papoti, Marcelo; Gobatto, Claudio A.

    2008-01-01

    The aim of this study was to validate critical frequency specific test (critf) for the estimation of the aerobic endurance in table tennis players. Methods: Eight male international-level table tennis players participated of this study. Specific tests were applied by using a mechanical ball thrower to control the intensity of the exercise. The critf was determined by applying three or four series of exercises to exhaustion (Tlim). The critf was evaluated by using lactate steady state test (90, 100, and 106 % of critf intensity). The other specific test was an incremental protocol used to determine the anaerobic threshold (AnTBI) and the onset of blood lactate accumulation (OBLA) using a ball thrower. Results: The critf (39.87 ± 3.31 balls·min-1) was not significantly different among AnTBI (48.11 ± 7.36 balls·min- 1) and OBLA3.5 (49.36 ± 12.04 balls·min-1) frequencies and it was correlated with AnTBI parameter (r = 0.78). At frequencies of the 90 and 100% of critf a dynamic equilibrium was verified in lactate concentration between the eighth and twentieth minutes. However, this dynamic equilibrium was not found at 106% intensity. Conclusion: The data indicate that in table tennis the critf model can be used for measuring the aerobic endurance. Key pointsIn table tennis is need the use of a specific protocol for evaluation of the aerobic endurance.The critical frequency test in table tennis seems to represent the intensity of maximal equilibrium of lactatemia.The critical frequency test can be used for measuring table tennis aerobic endurance through specific protocol. PMID:24149951

  1. When activity requires breaking up: LEKTI proteolytic activation cascade for specific proteinase inhibition.

    PubMed

    Furio, Laetitia; Hovnanian, Alain

    2011-11-01

    Lymphoepithelial Kazal-type related inhibitor (LEKTI) is a multidomain proteinase inhibitor whose defective expression causes Netherton syndrome (NS). LEKTI is encoded by SPINK5, which is also a susceptibility gene for atopic disease. In this issue, Fortugno et al. report an elegant and thorough study of the LEKTI proteolytic activation process in which they identify the precise nature of the cleavage sites used and the bioactive fragments generated. They propose a proteolytic activation model in human skin and confirm differential inhibition of kallikrein (KLK) 5, 7, and 14 by the major physiological LEKTI fragments. They show that these bioactive fragments inhibit KLK-mediated proteolysis of desmoglein 1 (DSG1) and suggest a fine-tuned inhibition process controlling target serine proteinase (SP) activity. PMID:21997416

  2. Differential Neural Activity during Search of Specific and General Autobiographical Memories elicited by Musical Cues

    PubMed Central

    Ford, Jaclyn Hennessey; Addis, Donna Rose; Giovanello, Kelly S.

    2011-01-01

    Previous neuroimaging studies that have examined autobiographical memory specificity have utilized retrieval cues associated with prior searches of the event, potentially changing the retrieval processes being investigated. In the current study, musical cues were used to naturally elicit memories from multiple levels of specificity (i.e., lifetime period, general event, and event-specific). Sixteen young adults participated in a neuroimaging study in which they retrieved autobiographical memories associated with musical cues. These musical cues led to the retrieval of highly emotional memories that had low levels of prior retrieval. Retrieval of all autobiographical memory levels was associated with activity in regions in the autobiographical memory network, specifically the ventromedial prefrontal cortex, posterior cingulate, and right medial temporal lobe. Owing to the use of music, memories from varying levels of specificity were retrieved, allowing for comparison of event memory and abstract personal knowledge, as well as comparison of specific and general event memory. Dorsolateral and dorsomedial prefrontal regions were engaged during event retrieval relative to personal knowledge retrieval, and retrieval of specific event memories was associated with increased activity in the bilateral medial temporal lobe and dorsomedial prefrontal cortex relative to retrieval of general event memories. These results suggest that the initial search processes for memories of different specificity levels preferentially engage different components of the autobiographical memory network. The potential underlying causes of these neural differences are discussed. PMID:21600227

  3. Relative position and pose measurement approach of specific operation site of space non-cooperative target

    NASA Astrophysics Data System (ADS)

    Chen, Feng; Zhou, Ying; Li, Ronghua; Huang, Jianming

    2015-12-01

    In order to achieve the rendezvous and capture of the space non-cooperative target, the relative position and pose measurement of non-cooperative target must be resolved. Since the marker is not installed into the non-cooperative target and there is no inter satellite link to transfer the information, so it is very difficult to measure the relative position and pose measurement of non-cooperative target. The solar array connecting frame of non-cooperative targets have their characters and are easy to capture, so the position and pose measurement of specific operation site of non-cooperative target based on stereo vision has been studied in this paper. The method composed of image acquiring, image filtering, edge detection, feature extraction and relative pose measurement. Finally, the relative position and attitude parameters of the solar wing connection were obtained and provided to the control system. The results of simulation and ground verification show that the algorithm is accurate and effective, and can satisfy the technical requirements of the on orbit operation. The measurement approach can be used for engineering implementation.

  4. Site-specific monoubiquitination activates Ras by impeding GTPase-activating protein function

    PubMed Central

    Hobbs, G Aaron; Gunawardena, Harsha P; Baker, Rachael; Campbell, Sharon L

    2013-01-01

    KRas has recently been shown to be activated by monoubiquitination (mUb). Similar to oncogenic mutations, mUb of Ras at position 147 activates Ras by causing a defect in GTPase activating protein (GAP) function. To characterize the mechanism by which mUb impairs GAP-mediated downregulation of Ras, we made various modifications at position 147 of Ras and examined the impact on Ras sensitivity to GAP function. Whereas small modifications (iodoacetamide and glutathione) at position 147 of Ras do not affect GAP-mediated hydrolysis, ligation of Ras to UbG76C (native linker), UbX77C (one residue longer), and PDZ2 (with a native ubiquitin linker) was defective in GAP-mediated GTP hydrolysis. However, restoration of GAP activity was observed for Ras modified with the PDZ2 domain containing a shorter and stiffer linker region than ubiquitin. Therefore, the properties of the linker region dictate whether modification affects GAP-mediated hydrolysis, and our data indicate that the GAP defect requires a minimum linker length of 7 to 8 residues. PMID:24030601

  5. Inferring brain-computational mechanisms with models of activity measurements.

    PubMed

    Kriegeskorte, Nikolaus; Diedrichsen, Jörn

    2016-10-01

    High-resolution functional imaging is providing increasingly rich measurements of brain activity in animals and humans. A major challenge is to leverage such data to gain insight into the brain's computational mechanisms. The first step is to define candidate brain-computational models (BCMs) that can perform the behavioural task in question. We would then like to infer which of the candidate BCMs best accounts for measured brain-activity data. Here we describe a method that complements each BCM by a measurement model (MM), which simulates the way the brain-activity measurements reflect neuronal activity (e.g. local averaging in functional magnetic resonance imaging (fMRI) voxels or sparse sampling in array recordings). The resulting generative model (BCM-MM) produces simulated measurements. To avoid having to fit the MM to predict each individual measurement channel of the brain-activity data, we compare the measured and predicted data at the level of summary statistics. We describe a novel particular implementation of this approach, called probabilistic representational similarity analysis (pRSA) with MMs, which uses representational dissimilarity matrices (RDMs) as the summary statistics. We validate this method by simulations of fMRI measurements (locally averaging voxels) based on a deep convolutional neural network for visual object recognition. Results indicate that the way the measurements sample the activity patterns strongly affects the apparent representational dissimilarities. However, modelling of the measurement process can account for these effects, and different BCMs remain distinguishable even under substantial noise. The pRSA method enables us to perform Bayesian inference on the set of BCMs and to recognize the data-generating model in each case.This article is part of the themed issue 'Interpreting BOLD: a dialogue between cognitive and cellular neuroscience'. PMID:27574316

  6. Goal-directed behavior under emotional distraction is preserved by enhanced task-specific activation.

    PubMed

    Wessa, Michèle; Heissler, Janine; Schönfelder, Sandra; Kanske, Philipp

    2013-03-01

    Despite the distracting effects of emotional stimuli on concurrent task performance, humans are able to uphold goal-directed behavior. Here, we investigated the hypothesis that this effect is due to the enhanced recruitment of task-specific neural resources. In a two-step functional magnetic resonance imaging study, we first localized those areas involved in mental arithmetics by contrasting arithmetic problems with a number detection task. The resulting activation maps were then used as masks in a second experiment that compared the effects of neutral and emotional distracter images on mental arithmetics. We found increased response times in the emotional distracter condition, accompanied by enhanced activation in task-specific areas, including superior parietal cortex, dorsolateral and dorsomedial prefrontal cortex. This activation increase correlated with larger behavioral impairment through emotional distraction. Similar error rates in both conditions indicate that cognitive task performance is preserved through enhanced recruitment of task-specific neural resources when emotional distracter stimuli are present. PMID:22302842

  7. Future of low specific activity molybdenum-99/technetium-99m generator.

    PubMed

    Mushtaq, A

    2012-10-01

    In last few years, the shortage of molybdenum-99 (99Mo) was felt in the developed and developing countries hospitals, where diagnostic nuclear medicine is practiced. To overcome the shortage of 99Mo various routes of its production by accelerators and reactors generating low and high specific activity products have been planned. High specific activity 99Mo obtained by fission of uranium-235 (235U) has completely dominated in the manufacturing of technetium-99m (99mTc) generators in last 3-4 decades, but due to proliferation and dirty bomb, issues non fission routes of 99Mo production are emphasized. Future of low specific activity 99Mo is discussed. PMID:22642420

  8. Earthworm Biomass Measurement: A Science Activity for Middle School.

    ERIC Educational Resources Information Center

    Haskett, Jonathan; Levine, Elissa; Carey, Pauline B.; Niepold III, Frank

    2000-01-01

    Describes an activity on biomass measurement which, in this case, is the weight of a group of living things in a given area. The earthworm activity gives students a greater understanding of ecology, practical math applications, and the scientific method. (ASK)

  9. Seasonality in Children's Pedometer-Measured Physical Activity Levels

    ERIC Educational Resources Information Center

    Beighle, Aaron; Alderman, Brandon; Morgan, Charles F.; Le Masurier, Guy

    2008-01-01

    Seasonality appears to have an impact on children's physical activity levels, but equivocal findings demand more study in this area. With the increased use of pedometers in both research and practice, collecting descriptive data in various seasons to examine the impact of seasonality on pedometer-measured physical activity among children is…

  10. A Simple and Accurate Method for Measuring Enzyme Activity.

    ERIC Educational Resources Information Center

    Yip, Din-Yan

    1997-01-01

    Presents methods commonly used for investigating enzyme activity using catalase and presents a new method for measuring catalase activity that is more reliable and accurate. Provides results that are readily reproduced and quantified. Can also be used for investigations of enzyme properties such as the effects of temperature, pH, inhibitors,…

  11. Should soil testing services measure soil biological activity?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Health of agricultural soils depends largely on conservation management to promote soil organic C accumulation. Total soil organic C changes slowly, but active fractions are more dynamic. A key indicator of healthy soil is potential biological activity, which could be measured rapidly with soil te...

  12. Thermal Expansion, Specific Heat and Magnetostriction Measurements on R-Copper

    NASA Astrophysics Data System (ADS)

    Chien, Teh-Shih

    The RCu (R = Gd, Tb, Dy and Ho) and R _2In (R = Gd and Tb) alloys have been systematically studied by thermal expansion, specific heat and magnetostriction measurements in order to investigate their magnetic and physical behaviors. GdCu and TbCu alloys undergo martensitic transformations at high and low temperatures. The Neel temperature of the GdCu alloy is 141.3 K from thermal expansion measurements. The Neel temperature T_{rm N} and martensitic transformation temperature M _{rm s} are 113.6 K and 116 K, respectively, for TbCu alloy. This is the first study to distinguish T_{rm N} from M_{rm s} using thermal expansion and specific heat measurements as well as a large thermal hysteresis. Both GdCu and TbCu alloys have a first-order structural transformation and a second-order magnetic phase transition. DyCu alloy has T_{rm N} = 60.5 K. The magnetic specific heat, C_{ rm m}, is a function of T^3 which obeys spin wave theory. HoCu alloy has T _{rm N} = 26 K and a spin reorientation at 14.1 K. YCu alloy has a Debye temperature of 230 K and C_{rm e} = 0.002T J/moleK. The Debye temperature is 160 K for all RCu alloys except for the DyCu alloy which has theta = 150 K. Gd_2In alloy has T _{rm N} = 97 K and T _{rm c} = 190.3 K which are associated with the antiferromagnetic and ferromagnetic transitions, respectively, from thermal expansion and magnetostriction measurements. Gd_2In alloy is a metamagnet with a critical magnetic field H = 8 kOe. Volume magnetostriction, omega_{rm V} is a function of H^{2 over3} in the ferromagnetic state. omega_{rm v} is a function of H^2, as expected, in the antiferromagnetic and paramagnetic states. The Curie temperature is 167.5 K for Tb_2In, as given by the thermal expansion and specific heat measurements. omega_{rm v} is a function of H in the ferromagnetic state. omega_{rm v} is a function of H^2, as expected, in the paramagnetic state.

  13. LOFT experimental measurements uncertainty analyses. Volume XX. Fluid-velocity measurement using pulsed-neutron activation

    SciTech Connect

    Lassahn, G.D.; Taylor, D.J.N.

    1982-08-01

    Analyses of uncertainty components inherent in pulsed-neutron-activation (PNA) measurements in general and the Loss-of-Fluid-Test (LOFT) system in particular are given. Due to the LOFT system's unique conditions, previously-used techniques were modified to make the volocity measurement. These methods render a useful, cost-effective measurement with an estimated uncertainty of 11% of reading.

  14. Active transport, substrate specificity, and methylation of Hg(II) in anaerobic bacteria

    SciTech Connect

    Schasfer, Jeffra; Rocks, Sara; Zheng, Wang; Liang, Liyuan; Gu, Baohua; Morel, Francois M

    2011-01-01

    The formation of methylmercury (MeHg), which is biomagnified in aquatic food chains and poses a risk to human health, is effected by some iron- and sulfate-reducing bacteria (FeRB and SRB) in anaerobic environments. However, very little is known regarding the mechanism of uptake of inorganic Hg by these organisms, in part because of the inherent difficulty in measuring the intracellular Hg concentration. By using the FeRB Geobacter sulfurreducens and the SRB Desulfovibrio desulfuricans ND132 as model organisms, we demonstrate that Hg(II) uptake occurs by active transport. We also establish that Hg(II) uptake by G. sulfurreducens is highly dependent on the characteristics of the thiols that bind Hg(II) in the external medium, with some thiols promoting uptake and methylation and others inhibiting both. The Hg(II) uptake system of D. desulfuricans has a higher affinity than that of G. sulfurreducens and promotes Hg methylation in the presence of stronger complexing thiols. We observed a tight coupling between Hg methylation and MeHg export from the cell, suggesting that these two processes may serve to avoid the build up and toxicity of cellular Hg. Our results bring up the question of whether cellular Hg uptake is specific for Hg(II) or accidental, occurring via some essential metal importer. Our data also point at Hg(II) complexation by thiols as an important factor controlling Hg methylation in anaerobic environments.

  15. The retrieval of self-defining memories is associated with the activation of specific working selves.

    PubMed

    Çili, Soljana; Stopa, Lusia

    2015-01-01

    This article presents two studies that investigated the impact of the retrieval of self-defining memories on individuals' sense of self. Participants recalled positive and/or negative self-defining memories, rated memory characteristics and completed measures focusing on different self-aspects. Study 1 found that participants reported higher state self-esteem after recalling a positive memory than after recalling a negative one. They also reported lower negative self-consistency and higher state self-concept clarity and positive self-consistency, but this result became non-significant after controlling for state self-esteem. Study 2 found that participants reported higher state self-esteem, a marginally higher proportion of recreation/exploration, goals and a marginally lower proportion of achievement goals after recalling a positive memory than after recalling a negative one. They also reported a higher proportion of self-cognitions referring to emotional states after recalling memories from which they had not abstracted meaning than after recalling memories from which they had done this. These findings suggest that the retrieval of vivid, emotional and highly self-relevant memories may be accompanied by the activation of specific self-representations or working selves. They also suggest that the experience of memory-related intrusive images may temporarily influence individuals' sense of self. The implications of these findings for clinical practice are discussed. PMID:24528183

  16. Measurement of action spectra of light-activated processes

    NASA Astrophysics Data System (ADS)

    Ross, Justin; Zvyagin, Andrei V.; Heckenberg, Norman R.; Upcroft, Jacqui; Upcroft, Peter; Rubinsztein-Dunlop, Halina H.

    2006-01-01

    We report on a new experimental technique suitable for measurement of light-activated processes, such as fluorophore transport. The usefulness of this technique is derived from its capacity to decouple the imaging and activation processes, allowing fluorescent imaging of fluorophore transport at a convenient activation wavelength. We demonstrate the efficiency of this new technique in determination of the action spectrum of the light mediated transport of rhodamine 123 into the parasitic protozoan Giardia duodenalis.

  17. The risks and consequences from the transport of low specific activity materials by truck

    SciTech Connect

    Finley, N.C.; McClure, J.D.; Cashwell, J.W.; Ostmeyer, R.O.; Wangler, M.

    1989-01-01

    The packaging and transport category of low specific activity (LSA) material was conceived for radioactive materials (RAM) that were considered inherently safe for transport. Such materials could be transported in ''strong, tight'' packages. The primary concern for the radionuclide content in LSA materials was associated with the potential for inhalation of particulate radioactive material. Thus, the specifications for the category in terms of specific activity were structured to preclude excessive inhalation hazard. The current version of the International Atomic Energy Agency (IAEA) regulations maintains the inhalation-related limits but also shows concern that the restrictions on the LSA category may not preclude an excessive external radiation hazard for gamma-emitting materials. This paper presents the results of an investigation of the potential for such excessive external radiation hazards, particularly for LSA materials with specific activity levels near the current regulatory limit. Analyses are discussed that were performed to evaluate the potential radiological impacts of highway accidents leading to the release of high radiation level-low specific activity (HRL-LSA) materials from their packagings. The results of the analyses are intended to provide a basis for evaluating restrictions on the quantity of gamma-emitting radionuclides that can be contained in an LSA shipment. 8 refs., 2 tabs.

  18. Direct measurements of mass-specific optical cross sections of single-component aerosol mixtures.

    PubMed

    Radney, James G; Ma, Xiaofei; Gillis, Keith A; Zachariah, Michael R; Hodges, Joseph T; Zangmeister, Christopher D

    2013-09-01

    The optical properties of atmospheric aerosols vary widely, being dependent upon particle composition, morphology, and mixing state. This diversity and complexity of aerosols motivates measurement techniques that can discriminate and quantify a variety of single- and multicomponent aerosols that are both internally and externally mixed. Here, we present a new combination of techniques to directly measure the mass-specific extinction and absorption cross sections of laboratory-generated aerosols that are relevant to atmospheric studies. Our approach employs a tandem differential mobility analyzer, an aerosol particle mass analyzer, cavity ring-down and photoacoustic spectrometers, and a condensation particle counter. This suite of instruments enables measurement of aerosol particle size, mass, extinction and absorption coefficients, and aerosol number density, respectively. Taken together, these observables yield the mass-specific extinction and absorption cross sections without the need to model particle morphology or account for sample collection artifacts. Here we demonstrate the technique in a set of case studies which involve complete separation of aerosol by charge, separation of an external mixture by mass, and discrimination between particle types by effective density and single-scattering albedo. PMID:23875772

  19. Sensitivity of Force Specifications to the Errors in Measuring the Interface Force

    NASA Technical Reports Server (NTRS)

    Worth, Daniel

    1999-01-01

    Force-Limited Random Vibration Testing has been applied in the last several years at NASA/GSFC for various programs at the instrument and system level. Different techniques have been developed over the last few decades to estimate the dynamic forces that the test article under consideration will encounter in the operational environment. Some of these techniques are described in the handbook, NASA-HDBK-7004, and the monograph, NASA-RP-1403. A key element in the ability to perform force-limited testing is multi-component force gauges. This paper will show how some measurement and calibration errors in force gauges are compensated for w en tie force specification is calculated. The resulting notches in the acceleration spectrum, when a random vibration test is performed, are the same as the notches produced during an uncompensated test that has no measurement errors. The paper will also present the results of tests that were used to validate this compensation. Knowing that the force specification can compensate for some measurement errors allows tests to continue after force gauge failures or allows dummy gauges to be used in places that are inaccessible.

  20. Ion-selective self-referencing probes for measuring specific ion flux

    PubMed Central

    Reid, Brian

    2011-01-01

    The metal vibrating probe developed in the 1970s to measure electric current is sensitive down to the micro-Amp range, but detects only net current due to flow of multiple ions and is too large to measure from single cells. Electrophysiological techniques which use glass microelectrodes such as voltage clamping can be used on single cells but are also non-specific. Ion-selective probes are glass microelectrodes containing at their tip a small amount of ionophore permeable to a particular ion. The electrode is therefore sensitive to changes in concentration of this ion. If the probe tip is moved at low frequency between two points in a concentration gradient of this ion then the electrochemical potential of the solution inside the electrode fluctuates in proportion to the size of the ion gradient. This fluctuation is amplified and recorded and is used to calculate the actual ion flux using Fick's law of diffusion. In this mini-review we describe the technique of ion-selective self-referencing microelectrodes to measure specific ion fluxes. We discuss the development of the technique and describe in detail the methodology and present some representative results. PMID:22046453

  1. A beverage-specific measure of expectancies for malt liquor: development and initial testing.

    PubMed

    Collins, R Lorraine; Vincent, Paula C; Bradizza, Clara M; Kubiak, Audrey J; Falco, Diana L

    2011-12-01

    Malt liquor (ML) is a unique, high alcohol content beverage marketed to encourage heavy drinking. We developed the Malt Liquor Expectancy Questionnaire (MLEQ), a beverage-specific measure of alcohol expectancies, and examined its association with typical weekly ML use, typical weekly alcohol use, and alcohol problems. Forty positive and 40 negative expectancy items were administered to a sample of 639 young adults who regularly consumed ML. Exploratory and confirmatory factor analyses led to the development of the 30-item MLEQ. The MLEQ consists of two positive (i.e., Social Facilitation and Enjoyment, Enhanced Sexuality) and two negative factors (i.e., Aggression and Negative Consequences; Impairment and Physical Symptoms) that possess good internal consistency, test-retest reliability, and convergent validity. The psychometrically sound MLEQ contributes to the limited research on beverage-specific expectancies and heavy drinking. PMID:21928870

  2. Increasing cleavage specificity and activity of restriction endonuclease KpnI

    PubMed Central

    Vasu, Kommireddy; Nagamalleswari, Easa; Zahran, Mai; Imhof, Petra; Xu, Shuang-yong; Zhu, Zhenyu; Chan, Siu-Hong; Nagaraja, Valakunja

    2013-01-01

    Restriction enzyme KpnI is a HNH superfamily endonuclease requiring divalent metal ions for DNA cleavage but not for binding. The active site of KpnI can accommodate metal ions of different atomic radii for DNA cleavage. Although Mg2+ ion higher than 500 μM mediates promiscuous activity, Ca2+ suppresses the promiscuity and induces high cleavage fidelity. Here, we report that a conservative mutation of the metal-coordinating residue D148 to Glu results in the elimination of the Ca2+-mediated cleavage but imparting high cleavage fidelity with Mg2+. High cleavage fidelity of the mutant D148E is achieved through better discrimination of the target site at the binding and cleavage steps. Biochemical experiments and molecular dynamics simulations suggest that the mutation inhibits Ca2+-mediated cleavage activity by altering the geometry of the Ca2+-bound HNH active site. Although the D148E mutant reduces the specific activity of the enzyme, we identified a suppressor mutation that increases the turnover rate to restore the specific activity of the high fidelity mutant to the wild-type level. Our results show that active site plasticity in coordinating different metal ions is related to KpnI promiscuous activity, and tinkering the metal ion coordination is a plausible way to reduce promiscuous activity of metalloenzymes. PMID:23963701

  3. Transferrin Binding to Peripheral Blood Lymphocytes Activated by Phytohemagglutinin Involves a Specific Receptor

    PubMed Central

    Galbraith, Robert M.; Werner, Phillip; Arnaud, Philippe; Galbraith, Gillian M. P.

    1980-01-01

    Immunohistological studies have indicated that membrane sites binding transferrin are present upon activated human peripheral blood lymphocytes. In this study, we have investigated transferrin uptake in human lymphocytes exposed to phytohemagglutinin (PHA), by quantitative radiobinding and immunofluorescence in parallel. In stimulated lymphocytes, binding was maximal after a 30-min incubation, being greatest at 37°C, and greater at 22°C than at 4°C. Although some shedding and endocytosis of transferrin occurred at 22° and 37°C, these factors, and resulting synthesis of new sites, did not affect measurement of binding which was found to be saturable, reversible, and specific for transferrin (Ka 0.5-2.5 × 108 M−1). Binding was greater after a 48-h exposure to PHA than after 24 h, and was maximal at 66 h. Sequential Scatchard analysis revealed no significant elevation in affinity of interaction. However, although the total number of receptors increased, the proportion of cells in which binding of ligand was detected immunohistologically increased in parallel, and after appropriate correction, the cellular density of receptors remained relatively constant throughout (60,000-80,000 sites/cell). Increments in binding during the culture period were thus due predominantly to expansion of a population of cells bearing receptors. Similar differences in binding were apparent upon comparison of cells cultured in different doses of PHA, and in unstimulated cells binding was negligible. Transferrin receptors appear, therefore, to be readily detectable only upon lymphocytes that have been activated. Images PMID:6253523

  4. Sequence of the lid affects activity and specificity of Candida rugosa lipase isoenzymes.

    PubMed

    Brocca, Stefania; Secundo, Francesco; Ossola, Mattia; Alberghina, Lilia; Carrea, Giacomo; Lotti, Marina

    2003-10-01

    The fungus Candida rugosa produces multiple lipase isoenzymes (CRLs) with distinct differences in substrate specificity, in particular with regard to selectivity toward the fatty acyl chain length. Moreover, isoform CRL3 displays high activity towards cholesterol esters. Lipase isoenzymes share over 80% sequence identity but diverge in the sequence of the lid, a mobile loop that modulates access to the active site. In the active enzyme conformation, the open lid participates in the substrate-binding site and contributes to substrate recognition. To address the role of the lid in CRL activity and specificity, we substituted the lid sequences from isoenzymes CRL3 and CRL4 in recombinant rCRL1, thus obtaining enzymes differing only in this stretch of residues. Swapping the CRL3 lid was sufficient to confer to CRL1 cholesterol esterase activity. On the other hand, a specific shift in the chain-length specificity was not observed. Chimeric proteins displayed different sensitivity to detergents in the reaction medium. PMID:14500889

  5. Sequence of the lid affects activity and specificity of Candida rugosa lipase isoenzymes

    PubMed Central

    Brocca, Stefania; Secundo, Francesco; Ossola, Mattia; Alberghina, Lilia; Carrea, Giacomo; Lotti, Marina

    2003-01-01

    The fungus Candida rugosa produces multiple lipase isoenzymes (CRLs) with distinct differences in substrate specificity, in particular with regard to selectivity toward the fatty acyl chain length. Moreover, isoform CRL3 displays high activity towards cholesterol esters. Lipase isoenzymes share over 80% sequence identity but diverge in the sequence of the lid, a mobile loop that modulates access to the active site. In the active enzyme conformation, the open lid participates in the substrate-binding site and contributes to substrate recognition. To address the role of the lid in CRL activity and specificity, we substituted the lid sequences from isoenzymes CRL3 and CRL4 in recombinant rCRL1, thus obtaining enzymes differing only in this stretch of residues. Swapping the CRL3 lid was sufficient to confer to CRL1 cholesterol esterase activity. On the other hand, a specific shift in the chain-length specificity was not observed. Chimeric proteins displayed different sensitivity to detergents in the reaction medium. PMID:14500889

  6. Natural variation in Fc glycosylation of HIV-specific antibodies impacts antiviral activity

    PubMed Central

    Ackerman, Margaret E.; Crispin, Max; Yu, Xiaojie; Baruah, Kavitha; Boesch, Austin W.; Harvey, David J.; Dugast, Anne-Sophie; Heizen, Erin L.; Ercan, Altan; Choi, Ickwon; Streeck, Hendrik; Nigrovic, Peter A.; Bailey-Kellogg, Chris; Scanlan, Chris; Alter, Galit

    2013-01-01

    While the induction of a neutralizing antibody response against HIV remains a daunting goal, data from both natural infection and vaccine-induced immune responses suggest that it may be possible to induce antibodies with enhanced Fc effector activity and improved antiviral control via vaccination. However, the specific features of naturally induced HIV-specific antibodies that allow for the potent recruitment of antiviral activity and the means by which these functions are regulated are poorly defined. Because antibody effector functions are critically dependent on antibody Fc domain glycosylation, we aimed to define the natural glycoforms associated with robust Fc-mediated antiviral activity. We demonstrate that spontaneous control of HIV and improved antiviral activity are associated with a dramatic shift in the global antibody-glycosylation profile toward agalactosylated glycoforms. HIV-specific antibodies exhibited an even greater frequency of agalactosylated, afucosylated, and asialylated glycans. These glycoforms were associated with enhanced Fc-mediated reduction of viral replication and enhanced Fc receptor binding and were consistent with transcriptional profiling of glycosyltransferases in peripheral B cells. These data suggest that B cell programs tune antibody glycosylation actively in an antigen-specific manner, potentially contributing to antiviral control during HIV infection. PMID:23563315

  7. Catalase activity of different Candida species after exposition to specific antiserum

    PubMed Central

    Miyasaka, Natália R.S.; Unterkircher, Carmelinda S.; Shimizu, Mario T.

    2008-01-01

    Antisera were developed in rabbits after challenge with intracellular antigens of Candida albicans, C. tropicalis and C. parapsilosis. Microorganism catalase has been correlated with virulence, resistance to drugs and immunogenicity. The intracellular catalase is consistently present in strains of Candida and in this paper, the enzyme activity was analysed by PAGE after exposition to antisera. The catalases of C. albicans, C. parapsilosis and C. tropicalis were immunogenic and differed in their binding to specific antibodies raised in rabbits. Tests of cross-reactivity between different Candida species showed that when antiserum from C. albicans immunized rabbit was incubated with intracellular extracts of these three Candida species, the catalases activities were abolished. However, the antisera from C. parapsilosis or C. tropicalis immunized rabbits did not affect the catalase activity of C. albicans; the enzyme of C. albicans was inactivated only by the antiserum to the catalase of own C. albicans. The antiserum to the catalase of C. tropicalis was species-specific and did not cross-react with catalases of C. albicans and C. parapsilosis. The activities of Aspergillus niger and bovine catalases were not affected by the antiserum from any Candida immunized rabbits. This report is a preliminary study of specific antisera that react against intracellular catalase of Candida sp. and neutralize the enzymatic activity. Further study is necessary to develop species-specific antibody once differences in the susceptibility of the Candida species to commonly used antifungal drugs make identification to the species level important. PMID:24031174

  8. Sandwich masking eliminates both visual awareness of faces and face-specific brain activity through a feedforward mechanism.

    PubMed

    Harris, Joseph A; Wu, Chien-Te; Woldorff, Marty G

    2011-01-01

    It is generally agreed that considerable amounts of low-level sensory processing of visual stimuli can occur without conscious awareness. On the other hand, the degree of higher level visual processing that occurs in the absence of awareness is as yet unclear. Here, event-related potential (ERP) measures of brain activity were recorded during a sandwich-masking paradigm, a commonly used approach for attenuating conscious awareness of visual stimulus content. In particular, the present study used a combination of ERP activation contrasts to track both early sensory-processing ERP components and face-specific N170 ERP activations, in trials with versus without awareness. The electrophysiological measures revealed that the sandwich masking abolished the early face-specific N170 neural response (peaking at ~170 ms post-stimulus), an effect that paralleled the abolition of awareness of face versus non-face image content. Furthermore, however, the masking appeared to render a strong attenuation of earlier feedforward visual sensory-processing signals. This early attenuation presumably resulted in insufficient information being fed into the higher level visual system pathways specific to object category processing, thus leading to unawareness of the visual object content. These results support a coupling of visual awareness and neural indices of face processing, while also demonstrating an early low-level mechanism of interference in sandwich masking. PMID:21669859

  9. Development of the Patient Activation Measure for mental health.

    PubMed

    Green, Carla A; Perrin, Nancy A; Polen, Michael R; Leo, Michael C; Hibbard, Judith H; Tusler, Martin

    2010-07-01

    Our objective was to adapt the physical health Patient Activation Measure (PAM) for use among people with mental health conditions (PAM-MH). Data came from three studies among people with chronic mental health conditions and were combined in Rasch analyses. The PAM-MH's psychometric properties equal those of the original 13-item PAM. Test-retest reliability and concurrent validity were good, and the PAM-MH showed sensitivity to change. The PAM-MH appears to be a reliable and valid measure of patient activation among individuals with mental health problems. It appears to have potential for use in assessing change in activation. PMID:19728074

  10. Laser acupuncture induced specific cerebral cortical and subcortical activations in humans.

    PubMed

    Siedentopf, Christian M; Koppelstaetter, Florian; Haala, Ilka Anna; Haid, Veronika; Rhomberg, Paul; Ischebeck, Anja; Buchberger, Waltraud; Felber, Stephan; Schlager, Andreas; Golaszewski, Stefan M

    2005-09-01

    As recent studies demonstrated, acupuncture can elicit activity in specific brain areas. This study aims to explore further the central effect using laser acupuncture. We investigated the cerebral effects of laser acupuncture at both acupoints GB43 with functional magnetic resonance imaging (fMRI). As a control condition the laser was mounted at the same acupoints but without application of laser stimulation. The group results showed significant brain activations within the thalamus, nucleus subthalamicus, nucleus ruber, the brainstem, and the Brodmann areas 40 and 22 for the acupuncture condition. No significant brain activations were observed within the placebo condition. The activations we observed were laser acupuncture-specific and predominantly ipsilateral. This supports the assumption that acupuncture is mediated by meridians, since meridians do not cross to the other side. Furthermore, we could show that laser acupuncture allows one to design a pure placebo condition. PMID:15990948

  11. EnMAP Specific BRDF Measurements and Vegetation Indices for Low-Growing Biomes in the Arctic Tundra

    NASA Astrophysics Data System (ADS)

    Buchhorn, M.; Heim, B.; van der Linden, S.

    2011-12-01

    Global warming is highest in the Arctic where the tundra regions are expected to undergo pronounced changes. Remote sensing can provide spatial and temporal data on variables linked to vegetation and ecosystem processes on global scale. Specifically, hyperspectral remote sensing data can relate to Vegetation Indices (VIs), Leaf Area Index (LAI) and the fraction of Photosynthetically active Radiation (fPAR). Currently, spaceborne hyperspectral imaging data is provided by the CHRIS/PROBA and the Hyperion programs. The Environmental Mapping and Analysis Program (EnMAP), a German hyperspectral mission, is the next step in this line and will provide high spectral resolution observations with a ground sampling distance of 30 meter. The continuous spectral sampling provided by imaging spectroscopy offers the possibility to develop robust algorithms for vegetation indices in low-growing tundra biomes. Since the EnMAP sensor has pointing capabilities, both spectral and directional reflection characteristics need to be taken into account. In order to examine the influence of the bidirectional reflectance distribution function (BRDF) on spectral variables such as the Vegetation Indices, LAI and fPAR, we developed the EnMAP specific field goniospectrometer, EyeSight. We took part at the summer field campaign of the Earth Cryosphere Institute (RU) in 2011 on the Yamal Peninsula, Western Siberia, Russia. Field spectroscopy, vegetation and biomass analysis, and field goniometer measurements under varying sun zenith angles and moisture conditions have been conducted. The spectral measurements and BRDF measurements of the low-growing tundra are presented and discussed.

  12. Improvements on Low Level Activity Gamma Measurements and X-ray Spectrometry at the CEA-MADERE Measurement Platform

    NASA Astrophysics Data System (ADS)

    Sergeyeva, Victoria; Domergue, Christophe; Destouches, Christophe; Girard, Jean Michel; Philibert, Hervé; Bonora, Jonathan; Thiollay, Nicolas; Lyoussi, Abdallah

    2016-02-01

    The CEA MADERE platform (Measurement Applied to DosimEtry in REactors) is a part of the Instrumentation Sensors and Dosimetry Laboratory (LDCI). This facility is dedicated to the specific activity measurements of solid and radioactive samples using Gamma and X-ray spectrometry. MADERE is a high-performance facility devoted to neutron dosimetry for experimental programs performed in CEA and for the irradiation surveillance programmes of PWR vessels. The MADERE platform is engaged in a continuous improvement process. Recently, two High Efficiency diodes have been integrated to the MADERE platform in order to manage the accurate low level activity measurements (few Bq per sample). This new equipment provides a good level of efficiency over the energy range from 60 keV to 2 MeV. The background continuum is reduced due to the use of a Ultra Low Background (ULB) lead shielding. Relative and absolute X-ray measurement techniques have been improved in order to facilitate absolute rhodium activity measurement (Rh103m) on solid samples. Additional efforts have been made to increase the accuracy of the relative niobium (Nb93m) activity measurement technique. The way of setting up an absolute measurement method for niobium is under investigation. After a presentation of the MADERE's measurement devices, this paper focuses on the technological options taken into account for the design of high efficiency measurement devices. Then, studies performed on X-ray measurement techniques are presented. Some details about the calculation of uncertainties and correction factors are also mentioned. Finally, future research and development axes are exposed.

  13. MCG measurement in the environment of active magnetic shield.

    PubMed

    Yamazaki, K; Kato, K; Kobayashi, K; Igarashi, A; Sato, T; Haga, A; Kasai, N

    2004-01-01

    MCG (Magnetocardiography) measurement by a SQUID gradiometer was attempted with only active magnetic shielding (active shielding). A three-axis-canceling-coil active shielding system, where three 16-10-16 turns-coil sets were put in the orthogonal directions, produces a homogeneous magnetic field in a considerable volume surrounding the center. Fluxgate sensors were used as the reference sensors of the system. The system can reduce environmental magnetic noise at low frequencies of less than a few Hz, at 50 Hz and at 150 Hz. Reducing such disturbances stabilizes biomagnetic measurement conditions for SQUIDs in the absence of magnetically shielded rooms (MSR). After filtering and averaging the measured MCG data by a first-order SQUID gradiometer with only the active shielding during the daytime, the QRS complex and T wave was clearly presented. PMID:16012640

  14. Solar irradiance measurements - Minimum through maximum solar activity

    NASA Technical Reports Server (NTRS)

    Lee, R. B., III; Gibson, M. A.; Shivakumar, N.; Wilson, R.; Kyle, H. L.; Mecherikunnel, A. T.

    1991-01-01

    The Earth Radiation Budget Satellite (ERBS) and the NOAA-9 spacecraft solar monitors were used to measure the total solar irradiance during the period October 1984 to December 1989. Decreasing trends in the irradiance measurements were observed as sunspot activity decreased to minimum levels in 1986; after 1986, increasing trends were observed as sunspot activity increased. The magnitude of the irradiance variability was found to be approximately 0.1 percent between sunspot minimum and maximum (late 1989). When compared with the 1984 to 1989 indices of solar magnetic activity, the irradiance trends appear to be in phase with the 11-year sunspot cycle. Both irradiance series yielded 1,365/sq Wm as the mean value of the solar irradiance, normalized to the mean earth/sun distance. The monitors are electrical substitution, active-cavity radiometers with estimated measurement precisions and accuracies of less than 0.02 and 0.2 percent, respectively.

  15. Measurement of actinides and strontium-90 in high activity waste

    SciTech Connect

    Maxwell, S.L. III; Nelson, M.R.

    1994-08-01

    The reliable measurement of trace radionuclides in high activity waste is important to support waste processing activities at SRS (F and H Area Waste Tanks, Extended Sludge Processing (ESP) and In-Tank precipitation (ITP) processing). Separation techniques are needed to remove high levels of gamma activity and alpha/beta interferences prior to analytical measurement. Using new extraction chromatographic resins from EiChrom Industries, Inc., the SRS Central Laboratory has developed new high speed separation methods that enable measurement of neptunium, thorium, uranium, plutonium, americium and strontium-90 in high activity waste solutions. Small particle size resin and applied vacuum are used to reduce analysis times and enhance column performance. Extraction chromatographic resins are easy to use and eliminate the generation of contaminated liquid organic waste.

  16. Sensitive and Specific Measurement of Minimal Residual Disease in Acute Lymphoblastic Leukemia

    PubMed Central

    Morley, Alexander A.; Latham, Sue; Brisco, Michael J.; Sykes, Pamela J.; Sutton, Rosemary; Hughes, Elizabeth; Wilczek, Vicki; Budgen, Bradley; van Zanten, Katrina; Kuss, Bryone J.; Venn, Nicola C.; Norris, Murray D.; Crock, Catherine; Storey, Colin; Revesz, Tamas; Waters, Keith

    2009-01-01

    A sensitive and specific quantitative real-time polymerase chain reaction method, involving three rounds of amplification with two allele-specific oligonucleotide primers directed against an rearrangement, was developed to quantify minimal residual disease (MRD) in B-lineage acute lymphoblastic leukemia (ALL). For a single sample containing 10 μg of good quality DNA, MRD was quantifiable down to approximately 10−6, which is at least 1 log more sensitive than current methods. Nonspecific amplification was rarely observed. The standard deviation of laboratory estimations was 0.32 log units at moderate or high levels of MRD, but increased markedly as the level of MRD and the number of intact marker gene rearrangements in the sample fell. In 23 children with ALL studied after induction therapy, the mean MRD level was 1.6 × 10−5 and levels ranged from 1.5 × 10−2 to less than 10−7. Comparisons with the conventional one-round quantitative polymerase chain reaction method on 29 samples from another 24 children who received treatment resulted in concordant results for 22 samples and discordant results for seven samples. The sensitivity and specificity of the method are due to the use of nested polymerase chain reaction, one segment-specific and two allele-specific oligonucleotide primers, and the use of a large amount of good quality DNA. This method may improve MRD-based decisions on treatment for ALL patients, and the principles should be applicable to DNA-based MRD measurements in other disorders. PMID:19324989

  17. Estimating Concentrations of Road-Salt Constituents in Highway-Runoff from Measurements of Specific Conductance

    USGS Publications Warehouse

    Granato, Gregory E.; Smith, Kirk P.

    1999-01-01

    Discrete or composite samples of highway runoff may not adequately represent in-storm water-quality fluctuations because continuous records of water stage, specific conductance, pH, and temperature of the runoff indicate that these properties fluctuate substantially during a storm. Continuous records of water-quality properties can be used to maximize the information obtained about the stormwater runoff system being studied and can provide the context needed to interpret analyses of water samples. Concentrations of the road-salt constituents calcium, sodium, and chloride in highway runoff were estimated from theoretical and empirical relations between specific conductance and the concentrations of these ions. These relations were examined using the analysis of 233 highwayrunoff samples collected from August 1988 through March 1995 at four highway-drainage monitoring stations along State Route 25 in southeastern Massachusetts. Theoretically, the specific conductance of a water sample is the sum of the individual conductances attributed to each ionic species in solution-the product of the concentrations of each ion in milliequivalents per liter (meq/L) multiplied by the equivalent ionic conductance at infinite dilution-thereby establishing the principle of superposition. Superposition provides an estimate of actual specific conductance that is within measurement error throughout the conductance range of many natural waters, with errors of less than ?5 percent below 1,000 microsiemens per centimeter (?S/cm) and ?10 percent between 1,000 and 4,000 ?S/cm if all major ionic constituents are accounted for. A semi-empirical method (adjusted superposition) was used to adjust for concentration effects-superposition-method prediction errors at high and low concentrations-and to relate measured specific conductance to that calculated using superposition. The adjusted superposition method, which was developed to interpret the State Route 25 highway-runoff records, accounts for

  18. Targeted disruption of cocaine-activated accumbens neurons prevents context-specific sensitization

    PubMed Central

    Koya, Eisuke; Golden, Sam A.; Harvey, Brandon K.; Guez, Danielle H.; Berkow, Alexander; Simmons, Danielle E.; Bossert, Jennifer M.; Nair, Sunila G.; Uejima, Jamie L.; Marin, Marcelo T.; Mitchell, Timothy; Farquhar, David; Ghosh, Sukhen; Mattson, Brandi J.; Hope, Bruce T.

    2009-01-01

    Learned associations between effects of abused drugs and the drug administration environment play important roles in drug addiction. Histochemical and electrophysiological studies suggest that these associations are encoded in sparsely distributed nucleus accumbens neurons that are selectively activated by drugs and drug-associated cues. Although correlations between accumbens neuronal activity and responsivity to drugs and drug cues have been observed, no technique exists for selectively manipulating these activated neurons and establishing their causal role in behavioral effects of drugs and drug cues. Here we describe a novel method, termed ‘Daun02-inactivation method’, that selectively inactivates a minority of neurons activated by cocaine in an environment repeatedly paired with cocaine to demonstrate a causal role for these activated neurons in context-specific cocaine-induced psychomotor sensitization in rats. This method provides a new tool to study causal roles of selectively activated neurons in behavioral effects of drugs and drug cues and in other learned behaviors. PMID:19620976

  19. DREAM Controls the On/Off Switch of Specific Activity-Dependent Transcription Pathways

    PubMed Central

    Mellström, Britt; Sahún, Ignasi; Ruiz-Nuño, Ana; Murtra, Patricia; Gomez-Villafuertes, Rosa; Savignac, Magali; Oliveros, Juan C.; Gonzalez, Paz; Kastanauskaite, Asta; Knafo, Shira; Zhuo, Min; Higuera-Matas, Alejandro; Errington, Michael L.; Maldonado, Rafael; DeFelipe, Javier; Jefferys, John G. R.; Bliss, Tim V. P.; Dierssen, Mara

    2014-01-01

    Changes in nuclear Ca2+ homeostasis activate specific gene expression programs and are central to the acquisition and storage of information in the brain. DREAM (downstream regulatory element antagonist modulator), also known as calsenilin/KChIP-3 (K+ channel interacting protein 3), is a Ca2+-binding protein that binds DNA and represses transcription in a Ca2+-dependent manner. To study the function of DREAM in the brain, we used transgenic mice expressing a Ca2+-insensitive/CREB-independent dominant active mutant DREAM (daDREAM). Using genome-wide analysis, we show that DREAM regulates the expression of specific activity-dependent transcription factors in the hippocampus, including Npas4, Nr4a1, Mef2c, JunB, and c-Fos. Furthermore, DREAM regulates its own expression, establishing an autoinhibitory feedback loop to terminate activity-dependent transcription. Ablation of DREAM does not modify activity-dependent transcription because of gene compensation by the other KChIP family members. The expression of daDREAM in the forebrain resulted in a complex phenotype characterized by loss of recurrent inhibition and enhanced long-term potentiation (LTP) in the dentate gyrus and impaired learning and memory. Our results indicate that DREAM is a major master switch transcription factor that regulates the on/off status of specific activity-dependent gene expression programs that control synaptic plasticity, learning, and memory. PMID:24366545

  20. Specific heat and magnetization measurements on the ferromagnetic phase transition of manganites and ruthenates

    NASA Astrophysics Data System (ADS)

    Kim, Dongkyun

    Ferromagnetic manganites and ruthenate samples were studied by specific heat and magnetization measurements. For specific heat measurements, a novel thin film micro-calorimeter was modified for bulk samples and an attachment-method using indium was developed. From the specific heat, using a chi2 minimization method, we obtained the critical exponent a. From magnetization measurements, we obtained critical exponents, beta, gamma, and delta, by the modified Arrot plot scheme. We found that our single crystal sample of La0.75Sr0.25 MnO3 has a typical second order ferromagnetic phase transition with critical exponents alpha = 0.05, beta = 0.4, gamma = 1.27 and delta = 4.1, in good agreement with the report on a single crystal of La0.7 Sr0.3MnO3. In contrast, measurement on a polycrystalline sample of La0.8Sr 0.2MnO3 was reported to have mean-field-like phase transition. For La1-xCaxMnO3 samples, we have studied x = 0.33 and x = 0.4 samples to verify different order of phase transitions. The critical exponents of the La0.6Ca0.4MnO3 sample were found to be alpha = 0.5, beta = 0.25, gamma = 1.03, and delta = 5.0. These values are in excellent agreement with the theoretical predictions of the tricritical point. The existence of the tricritical point is a strong evidence of the changing order of phase transitions. The evidence of the first order phase transition of La0.67Ca 0.33MnO3 include a meta-magnetic phase transition just above TC, a dependence of TC on applied field, and an extremely sharp specific heat peak at TC. The entropy change associated with the first order transition (2.3 J/molK) is estimated from volume expansion (1.8 J/molK) and the loss of magnetization (0.5 J/molK) via Clausius-Clapeyron equation. The discontinuous drop of DeltaM ˜ 1.3 muB, obtained from the Clausius-Clapeyron equation, is consistent with the magnetization measurements. We studied the critical behavior of a single crystal SrRuO3 sample and compared with resistivity data, drho

  1. Role of cerebral blood volume changes in brain specific-gravity measurements

    SciTech Connect

    Picozzi, P.; Todd, N.V.; Crockard, A.H.

    1985-05-01

    Cerebral blood volume (CBV) was calculated in gerbils from specific-gravity (SG) changes between normal and saline-perfused brains. Furthermore, changes in CBV were investigated during ischemia using carbon-14-labeled dextran (MW 70,000) as an intravascular marker. Both data were used to evaluate the possible error due to a change in CBV on the measurement of ischemic brain edema by the SG method. The methodological error found was 0.0004 for a 100% CBV change. This error is insignificant, being less than the standard deviation in the SG measured for the gerbil cortex. Thus, CBV changes are not responsible for the SG variations observed during the first phase of ischemia. These variations are better explained as an increase of brain water content during ischemia.

  2. Prostate involvement during sexually transmitted infections as measured by prostate-specific antigen concentration

    PubMed Central

    Sutcliffe, S; Nevin, R L; Pakpahan, R; Elliott, D J; Cole, S R; De Marzo, A M; Gaydos, C A; Isaacs, W B; Nelson, W G; Sokoll, L J; Zenilman, J M; Cersovsky, S B; Platz, E A

    2011-01-01

    Background: We investigated prostate involvement during sexually transmitted infections by measuring serum prostate-specific antigen (PSA) as a marker of prostate infection, inflammation, and/or cell damage in young, male US military members. Methods: We measured PSA before and during infection for 299 chlamydia, 112 gonorrhoea, and 59 non-chlamydial, non-gonococcal urethritis (NCNGU) cases, and 256 controls. Results: Chlamydia and gonorrhoea, but not NCNGU, cases were more likely to have a large rise (⩾40%) in PSA than controls (33.6%, 19.1%, and 8.2% vs 8.8%, P<0.0001, 0.021, and 0.92, respectively). Conclusion: Chlamydia and gonorrhoea may infect the prostate of some infected men. PMID:21792196

  3. Wounding Induces the Rapid and Transient Activation of a Specific MAP Kinase Pathway.

    PubMed Central

    Bogre, L.; Ligterink, W.; Meskiene, I.; Barker, P. J.; Heberle-Bors, E.; Huskisson, N. S.; Hirt, H.

    1997-01-01

    Mechanical injury in plants induces responses that are involved not only in healing but also in defense against a potential pathogen. To understand the intracellular signaling mechanism of wounding, we have investigated the involvement of protein kinases. Using specific antibodies, we showed that wounding alfalfa leaves specifically induces the transient activation of the p44MMK4 kinase, which belongs to the family of mitogen-activated protein kinases. Whereas activation of the MMK4 pathway is a post-translational process and was not blocked by [alpha]-amanitin and cycloheximide, inactivation depends on de novo transcription and translation of a protein factor(s). After wound-induced activation, the MMK4 pathway was subject to a refractory period of 25 min, during which time restimulation was not possible, indicating that the inactivation mechanism is only transiently active. After activation of the p44MMK4 kinase by wounding, transcript levels of the MMK4 gene increased, suggesting that the MMK4 gene may be a direct target of the MMK4 pathway. In contrast, transcripts of the wound-inducible MsWIP gene, encoding a putative proteinase inhibitor, were detected only several hours after wounding. Abscisic acid, methyl jasmonic acid, and electrical activity are known to mediate wound signaling in plants. However, none of these factors was able to activate the p44MMK4 kinase in the absence of wounding, suggesting that the MMK4 pathway acts independently of these signals. PMID:12237344

  4. Location-specific activation of the paraventricular nucleus of the hypothalamus by localized inflammation.

    PubMed

    Belevych, Natalya; Buchanan, Krystal; Chen, Qun; Bailey, Michael; Quan, Ning

    2010-10-01

    The existence of an immunological homunculus has been proposed, but evidence for location-specific response of the central nervous system to immunological stimulation is lacking. In this study, we show that inflammation induced by injection of casein into one of the causes c-fos expression in the paraventricular nucleus of the hypothalamus (PVN) in an asymmetrical manner: much stronger activation is always induced in the contralateral PVN. Unilateral sciatic nerve transection abolished the casein-induced PVN activation if casein was injected into the hindlimb with the nerve transection, but had no effect if casein was injected into the hindlimb with intact nerve innervation. Injection of casein into one the forelimbs also caused contralateral PNV activation. Further, stronger PVN activation was found in the anterior PVN after the forelimb injection, but in the posterior PVN after the hindlimb injection. Casein-induced PVN activation is absent in IL-1R1 KO, IL-6 KO, TNFα KO, and in C3H/HeJ (TLR4 mutant) animals. In comparison, injection of LPS, a systemic inflammagen, into one hindlimb induced bilateral PVN activation but injection of live Escherichia coli into one hindlimb induced contralateral PVN activation. These results support the notion that local inflammation may activate the PVN by neural routes in a location-specific manner. PMID:20570615

  5. Active-passive airborne ocean color measurement. II - Applications

    NASA Technical Reports Server (NTRS)

    Hoge, F. E.; Swift, R. N.; Yungel, J. K.

    1986-01-01

    Reported here for the first time is the use of a single airborne instrument to make concurrent measurements of oceanic chlorophyll concentration by (1) laser-induced fluorescence, (2) passive upwelling radiance, and (3) solar-induced chlorophyll fluorescence. Results from field experiments conducted with the NASA airborne oceanographic lidar (AOL) in the New York Bight demonstrate the capability of a single active-passive instrument to perform new and potentially important ocean color studies related to (1) active lidar validation of passive ocean color in-water algorithms, (2) chlorophyll a in vivo fluorescence yield variability, (3) calibration of active multichannel lidar systems, (4) effect of sea state on passive and active ocean color measurements, (5) laser/solar-induced chlorophyll fluorescence investigations, and (6) subsequent improvement of satellite-borne ocean color scanners. For validation and comparison purposes a separate passive ocean color sensor was also flown along with the new active-passive sensor during these initial field trials.

  6. Measuring the activities of higher organisms in activated sludge by means of mechanical shearing pretreatment and oxygen uptake rate.

    PubMed

    Hao, Xiaodi; Wang, Qilin; Cao, Yali; van Loosdrecht, Mark C M

    2010-07-01

    A pretreatment method was developed to assess the activities of higher organisms. The method is based on mechanical shearing to damage the large cells of the protozoan and metazoan community in activated sludge. The procedure was confirmed through experimentation to be effective in determining the activities of higher organisms by comparing oxygen uptake rates (OURs) before and after the higher organisms were eradicated. Shearing led to disintegration of flocs, which could be effectively reconstituted by centrifugation. The reconstitution of the sludge flocs was essential since otherwise the activity of the floc mass would be too high due to lack of diffusion limitation. Mechanical shearing had no influence on the morphology, quantity and specific activity of yeasts, and it was inferred that bacteria smaller than yeasts in size would also not be influenced by the applied shearing procedure. Moreover, the effect of filamentous organisms on the measured activities of higher organisms was experimentally demonstrated and analyzed, and determined to be so weak that it could be ignored. Based on these tests, five typical activated sludge processes were selected to measure the contribution of higher organisms to the original OUR. The measured activities of higher organisms ranged from 9.4 to 25.0% of the original OURs. PMID:20605188

  7. Promiscuous activity of ER glucosidase II discovered through donor specificity analysis of UGGT

    SciTech Connect

    Miyagawa, Atsushi; Totani, Kiichiro; Matsuo, Ichiro; Ito, Yukishige

    2010-12-17

    Research highlights: {yields} UGGT has a narrow donor specificity. {yields} UGGT gave several non-natural high-mannose-type glycans. {yields} G-II has a promiscuous activity as broad specificity hexosidase. -- Abstract: In glycoprotein quality control system in the endoplasmic reticulum (ER), UGGT (UDP-glucose:glycoprotein glucosyltransferase) and glucosidase II (G-II) play key roles. UGGT serves as a glycoprotein folding sensor by virtue of its unique specificity to glucosylate glycoproteins at incompletely folded stage. By using various UDP-Glc analogues, we first analyzed donor specificity of UGGT, which was proven to be rather narrow. However, marginal activity was observed with UDP-galactose and UDP-glucuronic acid as well as with 3-, 4- and 6-deoxy glucose analogues to give corresponding transfer products. Intriguingly, G-II smoothly converted all of them back to Man{sub 9}GlcNAc{sub 2}, providing an indication that G-II has a promiscuous activity as a broad specificity hexosidase.

  8. Activation energies of colloidal particle aggregation: towards a quantitative characterization of specific ion effects.

    PubMed

    Tian, Rui; Yang, Gang; Li, Hang; Gao, Xiaodan; Liu, Xinmin; Zhu, Hualing; Tang, Ying

    2014-05-21

    A quantitative description of specific ion effects is an essential and focused topic in colloidal and biological science. In this work, the dynamic light scattering technique was employed to study the aggregation kinetics of colloidal particles in the various alkali ion solutions with a wide range of concentrations. It indicated that the activation energies could be used to quantitatively characterize specific ion effects, which was supported by the results of effective hydrodynamic diameters, aggregation rates and critical coagulation concentrations. At a given concentration of 25 mmol L(-1), the activation energies for Li(+) are 1.2, 5.7, 28, and 126 times as much for Na(+), K(+), Rb(+), and Cs(+), respectively. Most importantly, the activation energy differences between two alkali cation species increase sharply with decrease of electrolyte concentrations, implying the more pronounced specific ion effects at lower concentrations. The dominant role of electrolyte cations during the aggregation of negatively charged colloidal particles was confirmed by alternative anions. Among the various theories, only the polarization effect can give a rational interpretation of the above specific ion effects, and this is substantially supported by the presence of strong electric fields from montmorillonite surfaces and its association mainly with electrolyte cations and montmorillonite particles. The classical induction theory, although with inclusion of electric field, requires significant corrections because it predicts an opposite trend to the experimentally observed specific ion effects. PMID:24603654

  9. Identification of an important motif that controls the activity and specificity of sugar transporters.

    PubMed

    Wang, Meng; Yu, Chenzhao; Zhao, Huimin

    2016-07-01

    Efficient glucose-xylose co-utilization is critical for economical biofuel production from lignocellulosic biomass. To enable glucose-xylose co-utilization, a highly active xylose specific transporter without glucose inhibition is desirable. However, our understanding of the structure-activity/specificity relationship of sugar transporters in general is limited, which hinders our ability to engineer xylose-specific transporters. In this study, via homology modeling and analysis of hexose sugar transporter HXT14 mutants, we identified a highly conserved YYX(T/P) motif that plays an important role in controlling the activity and specificity of sugar transporters. We demonstrated that mutating the two tyrosine residues of the motif to phenylalanine, respectively, improved glucose transport capacity across several different sugar transporters. Furthermore, we illustrated that by engineering the fourth position in the YYX(T/P) motif, the sugar specificity of transporters was significantly altered or even reversed towards xylose. Finally, using the engineered sugar transporter, genuine glucose-xylose co-fermentation was achieved. Biotechnol. Bioeng. 2016;113: 1460-1467. © 2016 Wiley Periodicals, Inc. PMID:26724683

  10. Specificity of indium-111 granulocyte scanning and fecal excretion measurement in inflammatory bowel disease--an autoradiographic study

    SciTech Connect

    Keshavarzian, A.; Price, Y.E.; Peters, A.M.; Lavender, J.P.; Wright, N.A.; Hodgson, H.J.

    1985-12-01

    The validity of /sup 111/In granulocyte scanning and fecal excretion measurement, as a reflection of loss of cells into the gastrointestinal tract, was studied using an autoradiographic technique in 11 patients in whom /sup 111/In granulocyte scan and colonoscopy were carried out simultaneously. /sup 111/In granulocytes were injected 1.5-4 hr prior to colonoscopy, and intraluminal fluid, mucosal brushings, and colonic biopsies were collected during the colonoscopy. In two patients with no histological evidence of inflammatory bowel disease, and four patients with clinically and histologically inactive inflammatory bowel disease, no /sup 111/Indium was detected in fluid, brushing, or biopsies. In five patients with active disease, 85% of the /sup 111/In activity in colonic fluid was precipitated by low-speed centrifugation. Autoradiography confirmed that the label remained attached to whole granulocytes in colonic fluid and mucosal brushings. Studies on biopsies, at intervals up to 4 1/2 hr following labeled granulocyte injection, demonstrated labeled polymorphonuclear neutrophils (PMNs) on the inflamed epithelial surface, with occasional cells in crypt abscesses by 110 min. We conclude that the techniques of /sup 111/In granulocyte scanning and fecal counting in patients with IBD are specifically measuring cell loss; labeled PMNs are capable of migrating through the gastrointestinal mucosa, in active disease, within 2 hr of administration.

  11. Specificity matters: criterion-related validity of contextualized and facet measures of conscientiousness in predicting college student performance.

    PubMed

    Woo, Sang Eun; Jin, Jing; LeBreton, James M

    2015-01-01

    To enhance the predictive validity of self-report personality measures, 2 distinct ways of increasing specificity of personality measures have been proposed in the literature-contextual specificity (i.e., providing a contextual referent) and content specificity (i.e., focusing on more specific constructs such as the Big Five facets). This study extends this line of research by examining whether there is an optimal way to configure, align, or integrate contextual and content specificity using measures of conscientiousness to predict college student success. A sample of 478 undergraduate students completed 4 measures of conscientiousness that varied in the level of content and contextual specificity. These forms of specificity were crossed to yield 4 distinct measures of conscientiousness. We then evaluated and compared the relative importance and the incremental importance of these different measures in the prediction of academic success. Superior predictive validity was found for both contextualized and facet measures of conscientiousness compared to a measure of global conscientiousness in predicting grade-point average and a broader behavioral criterion of student performance. When contextual and content specificity approaches were compared and combined, we observed the strongest predictive validity when the level of specificity is appropriately matched between predictor and criterion. PMID:25695753

  12. Small-Chamber Measurements of Chemical-Specific Emission Factors for Drywall

    SciTech Connect

    Maddalena, Randy; Russell, Marion; Apte, Michael G.

    2010-06-01

    Imported drywall installed in U.S. homes is suspected of being a source of odorous and potentially corrosive indoor pollutants. To support an investigation of those building materials by the Consumer Products Safety Commission (CPSC), Lawrence Berkeley National Laboratory (LBNL) measured chemical-specific emission factors for 30 samples of drywall materials. Emission factors are reported for 75 chemicals and 30 different drywall samples encompassing both domestic and imported stock and incorporating natural, synthetic, or mixed gypsum core material. CPSC supplied all drywall materials. First the drywall samples were isolated and conditioned in dedicated chambers, then they were transferred to small chambers where emission testing was performed. Four sampling and analysis methods were utilized to assess (1) volatile organic compounds, (2) low molecular weight carbonyls, (3) volatile sulfur compounds, and (4) reactive sulfur gases. LBNL developed a new method that combines the use of solid phase microextraction (SPME) with small emission chambers to measure the reactive sulfur gases, then extended that technique to measure the full suite of volatile sulfur compounds. The testing procedure and analysis methods are described in detail herein. Emission factors were measured under a single set of controlled environmental conditions. The results are compared graphically for each method and in detailed tables for use in estimating indoor exposure concentrations.

  13. On the reliable measurement of specific absorption rates and intrinsic loss parameters in magnetic hyperthermia materials

    NASA Astrophysics Data System (ADS)

    Wildeboer, R. R.; Southern, P.; Pankhurst, Q. A.

    2014-12-01

    In the clinical application of magnetic hyperthermia, the heat generated by magnetic nanoparticles in an alternating magnetic field is used as a cancer treatment. The heating ability of the particles is quantified by the specific absorption rate (SAR), an extrinsic parameter based on the clinical response characteristic of power delivered per unit mass, and by the intrinsic loss parameter (ILP), an intrinsic parameter based on the heating capacity of the material. Even though both the SAR and ILP are widely used as comparative design parameters, they are almost always measured in non-adiabatic systems that make accurate measurements difficult. We present here the results of a systematic review of measurement methods for both SAR and ILP, leading to recommendations for a standardised, simple and reliable method for measurements using non-adiabatic systems. In a representative survey of 50 retrieved datasets taken from published papers, the derived SAR or ILP was found to be more than 5% overestimated in 24% of cases and more than 5% underestimated in 52% of cases.

  14. Deriving content-specific measures of room acoustic perception using a binaural, nonlinear auditory model.

    PubMed

    van Dorp Schuitman, Jasper; de Vries, Diemer; Lindau, Alexander

    2013-03-01

    Acousticians generally assess the acoustic qualities of a concert hall or any other room using impulse response-based measures such as the reverberation time, clarity index, and others. These parameters are used to predict perceptual attributes related to the acoustic qualities of the room. Various studies show that these physical measures are not able to predict the related perceptual attributes sufficiently well under all circumstances. In particular, it has been shown that physical measures are dependent on the state of occupation, are prone to exaggerated spatial fluctuation, and suffer from lacking discrimination regarding the kind of acoustic stimulus being presented. Accordingly, this paper proposes a method for the derivation of signal-based measures aiming at predicting aspects of room acoustic perception from content specific signal representations produced by a binaural, nonlinear model of the human auditory system. Listening tests were performed to test the proposed auditory parameters for both speech and music. The results look promising; the parameters correlate with their corresponding perceptual attributes in most cases. PMID:23464027

  15. Chemical synthesis of nucleoside-gamma-[32P]triphosphates of high specific activity.

    PubMed

    Janecka, A; Panusz, H; Pankowski, J; Koziołkiewicz, W

    1980-01-01

    A simple chemical procedure for the preparation of four common ribonucleoside 5-gamma-[32P]triphosphates of high specific activity (up to 10 Ci/mmole) based on the condensation of orthophosphoric acid with the corresponding nucleoside 5-diphosphate in the presence of ethyl chloroformate as well as the methods of purification and identification of the products are described. PMID:7375446

  16. Are There Gender-Specific Risk Factors for Suicidal Activity among Patients with Schizophrenia and Depression?

    ERIC Educational Resources Information Center

    Kaplan, Kalman J.; Harrow, Martin; Faull, Robert N.

    2012-01-01

    Are there gender-specific risk factors for suicidal activity among patients with schizophrenia and depression? A total of 74 schizophrenia patients (51 men, 23 women) and 77 unipolar nonpsychotic depressed patients (26 men, 51 women) from the Chicago Follow-up Study were studied prospectively at 2 years posthospitalization and again at 7.5 years.…

  17. Specification for installation of the crew activity planning system coaxial cable communication system

    NASA Technical Reports Server (NTRS)

    Allen, M. A.; Roman, G. S.

    1979-01-01

    The specification used to install a broadband coaxial cable communication system to support remote terminal operations on the Crew Activity Planning system at the Lyndon B. Johnson Space Center are reported. The system supports high speed communications between a Harris Slash 8 computer and one or more Sanders Graphic 7 displays.

  18. The calculation of surface orbital energies for specific types of active sites on dispersed metal catalysts

    SciTech Connect

    Augustine, R.L.; Lahanas, K.M.; Cole, F.

    1992-11-01

    An angular overlap calculation has been used to determine the s, p, and d orbital energy levels of the different types of surface sites present on dispersed metal catalysts. These data can permit a Frontier Molecular Orbital treatment of specific site activities as long as the surface orbital availability for overlap with adsorbed substrates is considered along with its energy value and symmetry.

  19. The calculation of surface orbital energies for specific types of active sites on dispersed metal catalysts

    SciTech Connect

    Augustine, R.L.; Lahanas, K.M.; Cole, F.

    1992-01-01

    An angular overlap calculation has been used to determine the s, p, and d orbital energy levels of the different types of surface sites present on dispersed metal catalysts. These data can permit a Frontier Molecular Orbital treatment of specific site activities as long as the surface orbital availability for overlap with adsorbed substrates is considered along with its energy value and symmetry.

  20. Specific and Nonspecific Neural Activity during Selective Processing of Visual Representations in Working Memory

    ERIC Educational Resources Information Center

    Oh, Hwamee; Leung, Hoi-Chung

    2010-01-01

    In this fMRI study, we investigated prefrontal cortex (PFC) and visual association regions during selective information processing. We recorded behavioral responses and neural activity during a delayed recognition task with a cue presented during the delay period. A specific cue ("Face" or "Scene") was used to indicate which one of the two…

  1. Transportation impact analysis for the shipment of low specific activity nitric acid. Revisison 1

    SciTech Connect

    Green, J.R.

    1995-05-16

    This is in support of the Plutonium-Uranium Extraction (PUREX) Facility Low Specific Activity (LSA) Nitric Acid Shipment Environmental Assessment. It analyzes potential toxicological and radiological risks associated with transportation of PUREX Facility LSA Nitric Acid from the Hanford Site to Portsmouth VA, Baltimore MD, and Port Elizabeth NJ.

  2. Designing Class Activities to Meet Specific Core Training Competencies: A Developmental Approach

    ERIC Educational Resources Information Center

    Guth, Lorraine J.; McDonnell, Kelly A.

    2004-01-01

    This article presents a developmental model for designing and utilizing class activities to meet specific Association for Specialists in Group Work (ASGW) core training competencies for group workers. A review of the relevant literature about teaching group work and meeting core training standards is provided. The authors suggest a process by…

  3. Context Fear Learning Specifically Activates Distinct Populations of Neurons in Amygdala and Hypothalamus

    ERIC Educational Resources Information Center

    Trogrlic, Lidia; Wilson, Yvette M.; Newman, Andrew G.; Murphy, Mark

    2011-01-01

    The identity and distribution of neurons that are involved in any learning or memory event is not known. In previous studies, we identified a discrete population of neurons in the lateral amygdala that show learning-specific activation of a c-"fos"-regulated transgene following context fear conditioning. Here, we have extended these studies to…

  4. Measuring Alcohol-Specific Communication with Friends: Conceptualizing and Operationalizing Communication as Multidimensional.

    PubMed

    Kam, Jennifer A; Figueroa-Caballero, Andrea; Basinger, Erin D

    2016-06-01

    According to primary socialization theory, adolescents develop beliefs about alcohol by interacting with peers, as well as other socialization agents. Although communication is essential to this belief-formation process, few studies have identified the specific alcohol-related messages that adolescents exchange with their peers, and more specifically friends, that lead to certain anti- and/or pro-alcohol-related beliefs. Consequently, the goal of this study was to develop a multidimensional measure of alcohol-specific communication with friends. Based on survey data from 259 high school students, the results indicated that communication with friends involving warnings against drinking alcohol, disapproval of alcohol consumption, and making fun of others for drinking alcohol was negatively related to pro-alcohol beliefs and intentions. Communication with friends involving rumors, teasing each other about drinking alcohol, intentions to drink alcohol, different types of alcohol, experiences with alcohol, and talking about how many peers drink alcohol was positively related to pro-alcohol beliefs and intentions. PMID:26529419

  5. Development of the Abbott MATRIX Aero assay for the measurement of specific IgE.

    PubMed

    Lindberg, R E; Anawis, M A; Bailey, M; Mangat, D; Frank, P M; Hrusovsky, I G; Hooyman, L; Putterman, C; Defreese, J D

    1991-01-01

    An enzyme immunoassay has been developed for the quantitation of specific immunoglobulin E (IgE) in human serum to a panel of allergens. The assay system, called the Abbott MATRIX Aero, includes an instrument, reagents and test cell disposables. Each test cell contains fourteen airborne allergens individually localized on a nitrocellulose solid phase. Individual calibration curves for each allergen are established by the manufacturer and included in barcode form with each test kit. Stable factory calibration eliminates the need to establish a calibration curve with each assay run. The instrument automatically incubates, washes, and reads the test cell and prints each result, which ensures assay reproducibility and provides ease-of-use. Analysis of test results shows good agreement with another in vitro assay for specific IgE. The Abbott MATRIX Aero is a sensitive, reproducible and easy-to-use system for the measurement of specific IgE to a panel of fourteen allergens simultaneously using a single, small volume of serum. PMID:1806584

  6. Screening of DUB activity and specificity by MALDI-TOF mass spectrometry

    PubMed Central

    Ritorto, Maria Stella; Ewan, Richard; Perez-Oliva, Ana B.; Knebel, Axel; Buhrlage, Sara J.; Wightman, Melanie; Kelly, Sharon M.; Wood, Nicola T.; Virdee, Satpal; Gray, Nathanael S.; Morrice, Nicholas A.; Alessi, Dario R.; Trost, Matthias

    2014-01-01

    Deubiquitylases (DUBs) are key regulators of the ubiquitin system which cleave ubiquitin moieties from proteins and polyubiquitin chains. Several DUBs have been implicated in various diseases and are attractive drug targets. We have developed a sensitive and fast assay to quantify in vitro DUB enzyme activity using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Unlike other current assays, this method uses unmodified substrates, such as diubiquitin topoisomers. By analysing 42 human DUBs against all diubiquitin topoisomers we provide an extensive characterization of DUB activity and specificity. Our results confirm the high specificity of many members of the OTU and JAB/MPN/Mov34 metalloenzyme DUB families and highlight that all USPs tested display low linkage selectivity. We also demonstrate that this assay can be deployed to assess the potency and specificity of DUB inhibitors by profiling 11 compounds against a panel of 32 DUBs. PMID:25159004

  7. Screening of DUB activity and specificity by MALDI-TOF mass spectrometry.

    PubMed

    Ritorto, Maria Stella; Ewan, Richard; Perez-Oliva, Ana B; Knebel, Axel; Buhrlage, Sara J; Wightman, Melanie; Kelly, Sharon M; Wood, Nicola T; Virdee, Satpal; Gray, Nathanael S; Morrice, Nicholas A; Alessi, Dario R; Trost, Matthias

    2014-01-01

    Deubiquitylases (DUBs) are key regulators of the ubiquitin system which cleave ubiquitin moieties from proteins and polyubiquitin chains. Several DUBs have been implicated in various diseases and are attractive drug targets. We have developed a sensitive and fast assay to quantify in vitro DUB enzyme activity using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Unlike other current assays, this method uses unmodified substrates, such as diubiquitin topoisomers. By analysing 42 human DUBs against all diubiquitin topoisomers we provide an extensive characterization of DUB activity and specificity. Our results confirm the high specificity of many members of the OTU and JAB/MPN/Mov34 metalloenzyme DUB families and highlight that all USPs tested display low linkage selectivity. We also demonstrate that this assay can be deployed to assess the potency and specificity of DUB inhibitors by profiling 11 compounds against a panel of 32 DUBs. PMID:25159004

  8. Synthesis of high specific activity (1- sup 3 H) farnesyl pyrophosphate

    SciTech Connect

    Saljoughian, M.; Morimoto, H.; Williams, P.G.

    1991-08-01

    The synthesis of tritiated farnesyl pyrophosphate with high specific activity is reported. trans-trans Farnesol was oxidized to the corresponding aldehyde followed by reduction with lithium aluminium tritide (5%-{sup 3}H) to give trans-trans (1-{sup 3}H)farnesol. The specific radioactivity of the alcohol was determined from its triphenylsilane derivative, prepared under very mild conditions. The tritiated alcohol was phosphorylated by initial conversion to an allylic halide, and subsequent treatment of the halide with tris-tetra-n-butylammonium hydrogen pyrophosphate. The hydride procedure followed in this work has advantages over existing methods for the synthesis of tritiated farnesyl pyrophosphate, with the possibility of higher specific activity and a much higher yield obtained. 10 refs., 3 figs.

  9. Patient-specific left atrial wall-thickness measurement and visualization for radiofrequency ablation

    NASA Astrophysics Data System (ADS)

    Inoue, Jiro; Skanes, Allan C.; White, James A.; Rajchl, Martin; Drangova, Maria

    2014-03-01

    INTRODUCTION: For radiofrequency (RF) catheter ablation of the left atrium, safe and effective dosing of RF energy requires transmural left atrium ablation without injury to extra-cardiac structures. The thickness of the left atrial wall may be a key parameter in determining the appropriate amount of energy to deliver. While left atrial wall-thickness is known to exhibit inter- and intra-patient variation, this is not taken into account in the current clinical workflow. Our goal is to develop a tool for presenting patient-specific left atrial thickness information to the clinician in order to assist in the determination of the proper RF energy dose. METHODS: We use an interactive segmentation method with manual correction to segment the left atrial blood pool and heart wall from contrast-enhanced cardiac CT images. We then create a mesh from the segmented blood pool and determine the wall thickness, on a per-vertex basis, orthogonal to the mesh surface. The thickness measurement is visualized by assigning colors to the vertices of the blood pool mesh. We applied our method to 5 contrast-enhanced cardiac CT images. RESULTS: Left atrial wall-thickness measurements were generally consistent with published thickness ranges. Variations were found to exist between patients, and between regions within each patient. CONCLUSION: It is possible to visually determine areas of thick vs. thin heart wall with high resolution in a patient-specific manner.

  10. Experimental measurements of energy augmentation for mechanical circulatory assistance in a patient-specific Fontan model.

    PubMed

    Chopski, Steven G; Rangus, Owen M; Moskowitz, William B; Throckmorton, Amy L

    2014-09-01

    A mechanical blood pump specifically designed to increase pressure in the great veins would improve hemodynamic stability in adolescent and adult Fontan patients having dysfunctional cavopulmonary circulation. This study investigates the impact of axial-flow blood pumps on pressure, flow rate, and energy augmentation in the total cavopulmonary circulation (TCPC) using a patient-specific Fontan model. The experiments were conducted for three mechanical support configurations, which included an axial-flow impeller alone in the inferior vena cava (IVC) and an impeller with one of two different protective stent designs. All of the pump configurations led to an increase in pressure generation and flow in the Fontan circuit. The increase in IVC flow was found to augment pulmonary arterial flow, having only a small impact on the pressure and flow in the superior vena cava (SVC). Retrograde flow was neither observed nor measured from the TCPC junction into the SVC. All of the pump configurations enhanced the rate of power gain of the cavopulmonary circulation by adding energy and rotational force to the fluid flow. We measured an enhancement of forward flow into the TCPC junction, reduction in IVC pressure, and only minimally increased pulmonary arterial pressure under conditions of pump support. PMID:24404904

  11. Biocompatible click chemistry enabled compartment-specific pH measurement inside E. coli.

    PubMed

    Yang, Maiyun; Jalloh, Abubakar S; Wei, Wei; Zhao, Jing; Wu, Peng; Chen, Peng R

    2014-01-01

    Bioorthogonal reactions, especially the Cu(I)-catalysed azide-alkyne cycloaddition, have revolutionized our ability to label and manipulate biomolecules under living conditions. The cytotoxicity of Cu(I) ions, however, has hindered the application of this reaction in the internal space of living cells. By systematically surveying a panel of Cu(I)-stabilizing ligands in promoting protein labelling within the cytoplasm of Escherichia coli, we identify a highly efficient and biocompatible catalyst for intracellular modification of proteins by azide-alkyne cycloaddition. This reaction permits us to conjugate an environment-sensitive fluorophore site specifically onto HdeA, an acid-stress chaperone that adopts pH-dependent conformational changes, in both the periplasm and cytoplasm of E. coli. The resulting protein-fluorophore hybrid pH indicators enable compartment-specific pH measurement to determine the pH gradient across the E. coli cytoplasmic membrane. This construct also allows the measurement of E. coli transmembrane potential, and the determination of the proton motive force across its inner membrane under normal and acid-stress conditions. PMID:25236616

  12. Advanced Active-Magnetic-Bearing Thrust-Measurement System

    NASA Technical Reports Server (NTRS)

    Imlach, Joseph; Kasarda, Mary; Blumber, Eric

    2008-01-01

    An advanced thrust-measurement system utilizes active magnetic bearings to both (1) levitate a floating frame in all six degrees of freedom and (2) measure the levitation forces between the floating frame and a grounded frame. This system was developed for original use in measuring the thrust exerted by a rocket engine mounted on the floating frame, but can just as well be used in other force-measurement applications. This system offers several advantages over prior thrust-measurement systems based on mechanical support by flexures and/or load cells: The system includes multiple active magnetic bearings for each degree of freedom, so that by selective use of one, some, or all of these bearings, it is possible to test a given article over a wide force range in the same fixture, eliminating the need to transfer the article to different test fixtures to obtain the benefit of full-scale accuracy of different force-measurement devices for different force ranges. Like other active magnetic bearings, the active magnetic bearings of this system include closed-loop control subsystems, through which the stiffness and damping characteristics of the magnetic bearings can be modified electronically. The design of the system minimizes or eliminates cross-axis force-measurement errors. The active magnetic bearings are configured to provide support against movement along all three orthogonal Cartesian axes, and such that the support along a given axis does not produce force along any other axis. Moreover, by eliminating the need for such mechanical connections as flexures used in prior thrust-measurement systems, magnetic levitation of the floating frame eliminates what would otherwise be major sources of cross-axis forces and the associated measurement errors. Overall, relative to prior mechanical-support thrust-measurement systems, this system offers greater versatility for adaptation to a variety of test conditions and requirements. The basic idea of most prior active

  13. Comparative Validity of Physical Activity Measures in Older Adults

    PubMed Central

    COLBERT, LISA H.; MATTHEWS, CHARLES E.; HAVIGHURST, THOMAS C.; KIM, KYUNGMANN; SCHOELLER, DALE A.

    2012-01-01

    Purpose To compare the validity of various physical activity measures with doubly labeled water (DLW)–measured physical activity energy expenditure (PAEE) in free-living older adults. Methods Fifty-six adults aged ≥65 yr wore three activity monitors (New Lifestyles pedometer, ActiGraph accelerometer, and a SenseWear (SW) armband) during a 10-d free-living period and completed three different surveys (Yale Physical Activity Survey (YPAS), Community Health Activities Model Program for Seniors (CHAMPS), and a modified Physical Activity Scale for the Elderly (modPASE)). Total energy expenditure was measured using DLW, resting metabolic rate was measured with indirect calorimetry, the thermic effect of food was estimated, and from these, estimates of PAEE were calculated. The degree of linear association between the various measures and PAEE was assessed, as were differences in group PAEE, when estimable by a given measure. Results All three monitors were significantly correlated with PAEE (r = 0.48–0.60, P < 0.001). Of the questionnaires, only CHAMPS was significantly correlated with PAEE (r = 0.28, P = 0.04). Statistical comparison of the correlations suggested that the monitors were superior to YPAS and modPASE. Mean squared errors for all correlations were high, and the median PAEE from the different tools was significantly different from DLW for all but the YPAS and regression-estimated PAEE from the ActiGraph. Conclusions Objective devices more appropriately rank PAEE than self-reported instruments in older adults, but absolute estimates of PAEE are not accurate. Given the cost differential and ease of use, pedometers seem most useful in this population when ranking by physical activity level is adequate. PMID:20881882

  14. Assessing Built Environment Walkability using Activity-Space Summary Measures

    PubMed Central

    Tribby, Calvin P.; Miller, Harvey J.; Brown, Barbara B.; Werner, Carol M.; Smith, Ken R.

    2015-01-01

    There is increasing emphasis on active transportation, such as walking, in transportation planning as a sustainable form of mobility and in public health as a means of achieving recommended physical activity and better health outcomes. A research focus is the influence of the built environment on walking, with the ultimate goal of identifying environmental modifications that invite more walking. However, assessments of the built environment for walkability are typically at a spatially disaggregate level (such as street blocks) or at a spatially aggregate level (such as census block groups). A key issue is determining the spatial units for walkability measures so that they reflect potential walking behavior. This paper develops methods for assessing walkability within individual activity spaces: the geographic region accessible to an individual during a given walking trip. We first estimate street network-based activity spaces using the shortest path between known trip starting/ending points and a travel time budget that reflects potential alternative paths. Based on objective walkability measures of the street blocks, we use three summary measures for walkability within activity spaces: i) the average walkability score across block segments (representing the general level of walkability in the activity space); ii) the standard deviation (representing the walkability variation), and; iii) the network autocorrelation (representing the spatial coherence of the walkability pattern). We assess the method using data from an empirical study of built environment walkability and walking behavior in Salt Lake City, Utah, USA. We visualize and map these activity space summary measures to compare walkability among individuals’ trips within their neighborhoods. We also compare summary measures for activity spaces versus census block groups, with the result that they agree less than half of the time. PMID:27213027

  15. Measurement of the components of nonexercise activity thermogenesis.

    PubMed

    Levine, J; Melanson, E L; Westerterp, K R; Hill, J O

    2001-10-01

    Nonexercise activity thermogenesis (NEAT) accounts for the vast majority of nonresting metabolic rate and changes in NEAT-predicted susceptibility to fat gain with overfeeding. Measuring physical activity and its components in free-living humans has been a long-standing challenge. In this study, we combine information about lightweight sensors that capture data on body position and motion with laboratory measures of energy expenditure to calculate nonfidgeting NEAT. This measurement of nonfidgeting NEAT was compared with total NEAT measured in a room calorimeter in 11 healthy subjects. The measurement of nonfidgeting NEAT accounted for 85 +/- 9% of total NEAT measured in the room calorimeter. The intraclass correlation coefficient for the two methods was 0.86 (95% confidence interval 0.56, 0.96; P < 0.05). This suggests that 86% of the variance is attributable to between-subject variance and 14% to between-method disagreement. These instruments are applicable to free-living subjects; they are stand-alone, are lightweight, and allow normal daily activities. This novel technology has potential application for not only assessing NEAT but also tracking physical activity in free-living humans. PMID:11551842

  16. Controlling nuclear JAKs and STATs for specific gene activation by IFNγ

    PubMed Central

    Noon-Song, Ezra N.; Ahmed, Chulbul M.; Dabelic, Rea; Canton, Johnathan; Johnson, Howard M.

    2011-01-01

    We previously showed that gamma interferon (IFNγ) and its receptor subunit, IFNGR1, interacted with the promoter region of IFNγ-activated genes along with transcription factor STAT1α. Recent studies have suggested that activated Janus kinases pJAK2 and pJAK1 also played a role in gene activation by phosphorylation of histone H3 on tyrosine 41. This study addresses the question of the role of activated JAKs in specific gene activation by IFNγ. We carried out chromatin immunoprecipitation (ChIP) followed by PCR in IFNγ treated WISH cells and showed association of pJAK1, pJAK2, IFNGR1, and STAT1 on the same DNA sequence of the IRF-1 gene promoter. The β-actin gene, which is not activated by IFNγ, did not show this association. The movement of activated JAK to the nucleus and the IRF-1 promoter was confirmed by the combination of nuclear fractionation, confocal microscopy and DNA precipitation analysis using the biotinylated GAS promoter. Activated JAKs in the nucleus was associated with phosphorylated tyrosine 41 on histone H3 in the region of the GAS promoter. Unphosphorylated JAK2 was found to be constitutively present in the nucleus and was capable of undergoing activation in IFNγ treated cells, most likely via nuclear IFNGR1. Association of pJAK2 and IFNGR1 with histone H3 in IFNγ treated cells was demonstrated by histone H3 immunoprecipitation. Unphosphorylated STAT1 protein was associated with histone H3 of untreated cells. IFNγ treatment resulted in its disassociation and then re-association as pSTAT1. The results suggest a novel role for activated JAKs in epigenetic events for specific gene activation. PMID:21689637

  17. Should milk-specific IgE antibodies be measured in adults in primary care?

    PubMed Central

    Anthoni, Sari; Elg, Peter; Haahtela, Tari; Kolho, Kaija-Leena

    2008-01-01

    Objective To study the association of milk-IgE antibodies in serum to milk-related gastrointestinal symptoms in adults in primary care. Design Open clinical study. Setting Five outpatient clinics in primary care in Southern Finland. Subjects A total of 756 subjects who reported milk-related gastrointestinal symptoms in primary care and as controls 101 subjects with no such symptoms. Methods IgE values for specific food antigens were measured (Pharmacia CAP System) in a total of 857 subjects. All food screen-positive samples (>0.35 IU/l) were analysed further for IgE for untreated skimmed milk (milk-IgE) and for boiled milk. Those found positive for milk-IgE were invited for an open milk challenge test. Results Some 5.4% (46/857) of all subjects had a positive IgE antibody screen for food antigens. Of those with a positive food screen, 28% (13/46) had milk-IgE antibodies comprising 1.5% of the total group screened. The prevalence of milk-IgE was not statistically different between those with milk-related symptoms and those with no such symptoms. IgE antibodies for boiled milk were rare. All specific IgE antibody levels were low. Bloating was the only observed symptom in milk challenge tests. Conclusion IgE antibodies to cow's milk were relatively rare in the adult population and were not indicative of milk protein allergy. The observed IgE levels were low and did not correlate with subjective milk-related symptoms. The measurement of milk-specific IgE in adults should be discouraged in outpatient clinics. PMID:18609255

  18. Controlling nuclear JAKs and STATs for specific gene activation by IFN{gamma}

    SciTech Connect

    Noon-Song, Ezra N.; Ahmed, Chulbul M.; Dabelic, Rea; Canton, Johnathan; Johnson, Howard M.

    2011-07-08

    Highlights: {yields} Gamma interferon (IFN{gamma}) and its receptor subunit, IFNGR1, interact with the promoter region of IFN{gamma}-associated genes along with transcription factor STAT1{alpha}. {yields} We show that activated Janus kinases pJAK2 and pJAK1 also associate with IFNGR1 in the nucleus. {yields} The activated Janus kinases are responsible for phosphorylation of tyrosine 41 on histone H3, an important epigenetic event for specific gene activation. -- Abstract: We previously showed that gamma interferon (IFN{gamma}) and its receptor subunit, IFNGR1, interacted with the promoter region of IFN{gamma}-activated genes along with transcription factor STAT1{alpha}. Recent studies have suggested that activated Janus kinases pJAK2 and pJAK1 also played a role in gene activation by phosphorylation of histone H3 on tyrosine 41. This study addresses the question of the role of activated JAKs in specific gene activation by IFN{gamma}. We carried out chromatin immunoprecipitation (ChIP) followed by PCR in IFN{gamma} treated WISH cells and showed association of pJAK1, pJAK2, IFNGR1, and STAT1 on the same DNA sequence of the IRF-1 gene promoter. The {beta}-actin gene, which is not activated by IFN{gamma}, did not show this association. The movement of activated JAK to the nucleus and the IRF-1 promoter was confirmed by the combination of nuclear fractionation, confocal microscopy and DNA precipitation analysis using the biotinylated GAS promoter. Activated JAKs in the nucleus was associated with phosphorylated tyrosine 41 on histone H3 in the region of the GAS promoter. Unphosphorylated JAK2 was found to be constitutively present in the nucleus and was capable of undergoing activation in IFN{gamma} treated cells, most likely via nuclear IFNGR1. Association of pJAK2 and IFNGR1 with histone H3 in IFN{gamma} treated cells was demonstrated by histone H3 immunoprecipitation. Unphosphorylated STAT1 protein was associated with histone H3 of untreated cells. IFN

  19. Specific quorum sensing-disrupting activity (A QSI) of thiophenones and their therapeutic potential.

    PubMed

    Yang, Qian; Scheie, Anne Aamdal; Benneche, Tore; Defoirdt, Tom

    2015-01-01

    Disease caused by antibiotic resistant pathogens is becoming a serious problem, both in human and veterinary medicine. The inhibition of quorum sensing, bacterial cell-to-cell communication, is a promising alternative strategy to control disease. In this study, we determined the quorum sensing-disrupting activity of 20 thiophenones towards the quorum sensing model bacterium V. harveyi. In order to exclude false positives, we propose a new parameter (AQSI) to describe specific quorum sensing activity. AQSI is defined as the ratio between inhibition of quorum sensing-regulated activity in a reporter strain and inhibition of the same activity when it is independent of quorum sensing. Calculation of AQSI allowed to exclude five false positives, whereas the six most active thiophenones (TF203, TF307, TF319, TF339, TF342 and TF403) inhibited quorum sensing at 0.25 μM, with AQSI higher than 10. Further, we determined the protective effect and toxicity of the thiophenones in a highly controlled gnotobiotic model system with brine shrimp larvae. There was a strong positive correlation between the specific quorum sensing-disrupting activity of the thiophenones and the protection of brine shrimp larvae against pathogenic V. harveyi. Four of the most active quorum sensing-disrupting thiophenones (TF 203, TF319, TF339 and TF342) were considered to be promising since they have a therapeutic potential of at least 10. PMID:26647822

  20. Specific quorum sensing-disrupting activity (AQSI) of thiophenones and their therapeutic potential

    PubMed Central

    Yang, Qian; Aamdal Scheie, Anne; Benneche, Tore; Defoirdt, Tom

    2015-01-01

    Disease caused by antibiotic resistant pathogens is becoming a serious problem, both in human and veterinary medicine. The inhibition of quorum sensing, bacterial cell-to-cell communication, is a promising alternative strategy to control disease. In this study, we determined the quorum sensing-disrupting activity of 20 thiophenones towards the quorum sensing model bacterium V. harveyi. In order to exclude false positives, we propose a new parameter (AQSI) to describe specific quorum sensing activity. AQSI is defined as the ratio between inhibition of quorum sensing-regulated activity in a reporter strain and inhibition of the same activity when it is independent of quorum sensing. Calculation of AQSI allowed to exclude five false positives, whereas the six most active thiophenones (TF203, TF307, TF319, TF339, TF342 and TF403) inhibited quorum sensing at 0.25 μM, with AQSI higher than 10. Further, we determined the protective effect and toxicity of the thiophenones in a highly controlled gnotobiotic model system with brine shrimp larvae. There was a strong positive correlation between the specific quorum sensing-disrupting activity of the thiophenones and the protection of brine shrimp larvae against pathogenic V. harveyi. Four of the most active quorum sensing-disrupting thiophenones (TF 203, TF319, TF339 and TF342) were considered to be promising since they have a therapeutic potential of at least 10. PMID:26647822

  1. A sandwich ELISA for the conformation-specific quantification of the activated form of human Bax.

    PubMed

    Teijido, Oscar; Ganesan, Yogesh Tengarai; Llanos, Raul; Peton, Ashley; Urtecho, Jean-Baptiste; Soprani, Adauri; Villamayor, Aimee; Antonsson, Bruno; Manon, Stéphen; Dejean, Laurent

    2016-03-15

    Bcl-2 family proteins are critical regulators of mitochondrial outer membrane permeabilization (MOMP), which represents the point of no return of apoptotic cell death. The exposure of the Bax N-terminus at the mitochondria reflects Bax activation; and this activated configuration of the Bax protein is associated with MOMP. N-terminal exposure can be detected using specific monoclonal and/or polyclonal antibodies, and the onset of activated Bax has extensively been used as an early marker of apoptosis. The protocols of immunoprecipitation and/or immunocytochemistry commonly used to detect activated Bax are long and tedious, and allow semiquantification of the antigen at best. The sandwich ELISA protocol we developed has a 5 ng/mL detection limit and is highly specific for the activated conformation of Bax. This ELISA allows a rapid quantification of activated human Bax in whole cells and isolated mitochondria protein extracts. These properties grant this assay the potential to further clarify the prognostic and diagnostic value of activated Bax in disorders associated with deregulated apoptotic pathways such as degenerative diseases or cancer. PMID:26748144

  2. Mitochondria are required for antigen-specific T cell activation through reactive oxygen species signaling

    PubMed Central

    Sena, Laura A.; Li, Sha; Jairaman, Amit; Prakriya, Murali; Ezponda, Teresa; Hildeman, David A.; Wang, Chyung-Ru; Schumacker, Paul T.; Licht, Jonathan D.; Perlman, Harris; Bryce, Paul J.; Chandel, Navdeep S.

    2013-01-01

    SUMMARY It is widely appreciated that T cells increase glycolytic flux during activation, however the role of mitochondrial flux is unclear. Here we have shown that mitochondrial metabolism, in the absence of glucose metabolism, was sufficient to support interleukin-2 (IL-2) induction. Furthermore, we used mice with reduced mitochondrial reactive oxygen species (mROS) production in T cells (T-Uqcrfs−/− mice) to show that mitochondria are required for T cell activation to produce mROS for activation of nuclear factor of activated T cells (NFAT) and subsequent IL-2 induction. These mice could not induce antigen-specific expansion of T cells in vivo, however Uqcrfs1−/− T cells retained the ability to proliferate in vivo under lymphopenic conditions. This suggests that Uqcrfs1−/− T cells were not lacking bioenergetically, but rather lacked specific ROS-dependent signaling events needed for antigen-specific expansion. Thus, mitochondrial metabolism is a critical component of T cell activation through production of complex III ROS. PMID:23415911

  3. Sequence specific inhibition of human type II phospholipase A2 enzyme activity by phosphorothioate oligonucleotides.

    PubMed Central

    Bennett, C F; Chiang, M Y; Wilson-Lingardo, L; Wyatt, J R

    1994-01-01

    Phosphorothioate oligonucleotides were identified which directly inhibited human type II phospholipase A2 (PLA2) enzyme activity in a sequence specific manner. The minimum pharmacophore common to all oligonucleotides which inhibited PLA2 enzyme activity consisted of two sets of three or more consecutive guanosine residues in a row. These oligonucleotides appear to form G quartets resulting in the formation of oligonucleotide aggregates. Additionally, a phosphorothioate backbone was required to be effective inhibitors of type II PLA2. The activity of one oligodeoxynucleotide, IP 3196 (5'-GGGTGGGTATAGAAGGGCTCC-3') has been characterized in more detail. IP 3196 inhibited PLA2 enzyme activity when the substrate was presented in the form of a phospholipid bilayer but not when presented in the form of a mixed micelle with anionic detergents. Human type II PLA2 was 50-fold more sensitive to inhibition by IP 3196 than venom and pancreatic type I enzymes. These data demonstrate that phosphorothioate oligonucleotides can specifically inhibit human type II PLA2 enzyme activity in a sequence specific manner. PMID:8065936

  4. Determination of the specific alpha activity of thick sources with a large area ZnS(Ag) scintillation detector.

    PubMed

    Djurasević, M; Vukanac, I; Kandić, A; Nadderd, L; Milosević, Z; Radenković, M

    2007-01-01

    A method for determining the specific alpha activity of thick sources using a large area ZnS(Ag) scintillation detector is presented. In this method a quadratic relationship between the detector response and window thickness is assumed. This method provides a quick estimation of alpha activity in the sample, so it is an indicative method. The aim of this experimental work is to approve theoretical assumption and to develop a standard routine method for absolute alpha measurements of thick contaminated environmental sources. For this purpose reference material U(3)O(8) and spiked standards of soil were used. Measurements of contaminated soil samples from south Serbia showed the practical application of this method. PMID:17383779

  5. Elimination of dendritic spines with long-term memory is specific to active circuits

    PubMed Central

    Sanders, Jeff; Cowansage, Kiriana; Baumgärtel, Karsten; Mayford, Mark

    2012-01-01

    Structural changes in brain circuits active during learning are thought to be important for long-term memory storage. If these changes support long-term information storage they might be expected to be present at distant timepoints after learning, as well as specific to the circuit activated with learning, and sensitive to the contingencies of the behavioral paradigm. Here, we show such changes in the hippocampus as a result of contextual fear conditioning. There were significantly fewer spines specifically on active neurons of fear-conditioned mice. This spine loss did not occur in homecage mice or in mice exposed to the training context alone. Mice exposed to unpaired shocks showed a generalized reduction in spines. These learning-related changes in spine density could reflect a direct mechanism of encoding or alternately could reflect a compensatory adaptation to previously described enhancement in transmission due to glutamate receptor insertion. PMID:22956846

  6. Enhancement of neutralizing activity of influenza virus-specific antibodies by serum components.

    PubMed

    Mozdzanowska, Krystyna; Feng, Jingqi; Eid, Mark; Zharikova, Darya; Gerhard, Walter

    2006-09-01

    The role of serum components in enhancing virus neutralizing (VN) activity of influenza virus A/PR/8/34 hemagglutinin (HA)-specific MAbs in vitro was investigated. The degree of enhancement depended on the MAb's fine specificity and heavy chain isotype and on type of serum. Greatest enhancement (>100-fold) was seen with sera from immunodeficient mice that lacked serum immunoglobulin. At least two serum components were involved: C1q and a heat-resistant factor. C1q was mandatory for enhancement, and other components of the complement system were not required. C1q appeared to operate by improving MAb-mediated inhibition of virus attachment to host cells and was most effective with MAbs that inhibited virus attachment poorly on their own. The heat-resistant factor enhanced VN activity only in the presence of C1q and appeared to operate by enhancing VN activity at a post-attachment stage. PMID:16777168

  7. Methodological considerations for measuring spontaneous physical activity in rodents

    PubMed Central

    Perez-Leighton, Claudio E.; Billington, Charles J.; Kotz, Catherine M.

    2014-01-01

    When exploring biological determinants of spontaneous physical activity (SPA), it is critical to consider whether methodological factors differentially affect rodents and the measured SPA. We determined whether acclimation time, sensory stimulation, vendor, or chamber size affected measures in rodents with varying propensity for SPA. We used principal component analysis to determine which SPA components (ambulatory and vertical counts, time in SPA, and distance traveled) best described the variability in SPA measurements. We compared radiotelemetry and infrared photobeams used to measure SPA and exploratory activity. Acclimation time, sensory stimulation, vendor, and chamber size independently influenced SPA, and the effect was moderated by the propensity for SPA. A 24-h acclimation period prior to SPA measurement was sufficient for habituation. Principal component analysis showed that ambulatory and vertical measurements of SPA describe different dimensions of the rodent's SPA behavior. Smaller testing chambers and a sensory attenuation cubicle around the chamber reduced SPA. SPA varies between rodents purchased from different vendors. Radiotelemetry and infrared photobeams differ in their sensitivity to detect phenotypic differences in SPA and exploratory activity. These data highlight methodological considerations in rodent SPA measurement and a need to standardize SPA methodology. PMID:24598463

  8. Differential membrane-based nanocalorimeter for high-resolution measurements of low-temperature specific heat.

    PubMed

    Tagliati, S; Krasnov, V M; Rydh, A

    2012-05-01

    A differential, membrane-based nanocalorimeter for general specific heat studies of very small samples, ranging from 0.5 mg to sub-μg in mass, is described. The calorimeter operates over the temperature range from above room temperature down to 0.5 K. It consists of a pair of cells, each of which is a stack of heaters and thermometer in the center of a silicon nitride membrane, in total giving a background heat capacity less than 100 nJ/K at 300 K, decreasing to 10 pJ/K at 1 K. The device has several distinctive features: (i) The resistive thermometer, made of a Ge(1 - x)Au(x) alloy, displays a high dimensionless sensitivity ∣dlnR∕dlnT∣ ≳ 1 over the entire temperature range. (ii) The sample is placed in direct contact with the thermometer, which is allowed to self-heat. The thermometer can thus be operated at high dc current to increase the resolution. (iii) Data are acquired with a set of eight synchronized lock-in amplifiers measuring dc, 1st and 2nd harmonic signals of heaters and thermometer. This gives high resolution and allows continuous output adjustments without additional noise. (iv) Absolute accuracy is achieved via a variable-frequency-fixed-phase technique in which the measurement frequency is automatically adjusted during the measurements to account for the temperature variation of the sample heat capacity and the device thermal conductance. The performance of the calorimeter is illustrated by studying the heat capacity of a small Au sample and the specific heat of a 2.6 μg piece of superconducting Pb in various magnetic fields. PMID:22667657

  9. Validating the distribution of specific ventilation in healthy humans measured using proton MR imaging

    PubMed Central

    Asadi, Amran K.; Theilmann, Rebecca J.; Hopkins, Susan R.; Prisk, G. Kim; Darquenne, Chantal

    2014-01-01

    Specific ventilation imaging (SVI) uses proton MRI to quantitatively map the distribution of specific ventilation (SV) in the human lung, using inhaled oxygen as a contrast agent. To validate this recent technique, we compared the quantitative measures of heterogeneity of the SV distribution in a 15-mm sagittal slice of lung obtained in 10 healthy supine subjects, (age 37 ± 10 yr, forced expiratory volume in 1 s 97 ± 7% predicted) using SVI to those obtained in the whole lung from multiple-breath nitrogen washout (MBW). Using the analysis of Lewis et al. (Lewis SM, Evans JW, Jalowayski AA. J App Physiol 44: 416–423, 1978), the most likely distribution of SV from the MBW data was computed and compared with the distribution of SV obtained from SVI, after normalizing for the difference in tidal volume. The average SV was 0.30 ± 0.10 MBW, compared with 0.36 ± 0.10 SVI (P = 0.01). The width of the distribution, a measure of the heterogeneity, obtained using both methods was comparable: 0.51 ± 0.06 and 0.47 ± 0.08 in MBW and SVI, respectively (P = 0.15). The MBW estimated width of the SV distribution was 0.05 (10.7%) higher than that estimated using SVI, and smaller than the intertest variability of the MBW estimation [inter-MBW (SD) for the width of the SV distribution was 0.08 (15.8)%]. To assess reliability, SVI was performed twice on 13 subjects showing small differences between measurements of SV heterogeneity (typical error 0.05, 12%). In conclusion, quantitative estimations of SV heterogeneity from SVI are reliable and similar to those obtained using MBW, with SVI providing spatial information that is absent in MBW. PMID:24505099

  10. Position-specific measurement of oxygen isotope ratios in cellulose: Isotopic exchange during heterotrophic cellulose synthesis

    NASA Astrophysics Data System (ADS)

    Waterhouse, John S.; Cheng, Shuying; Juchelka, Dieter; Loader, Neil J.; McCarroll, Danny; Switsur, V. Roy; Gautam, Lata

    2013-07-01

    We describe the first reported method for the measurement of oxygen isotope ratios at each position in the glucose units of the cellulose molecule. The overall process comprises a series of synthetic organic sequences, by which α-cellulose is hydrolysed to glucose, and oxygen atoms at specific positions in the glucose molecule are removed in samples of benzoic acid for measurement of δ18O. Values of δ18O at specific positions in cellulose are calculated from these δ18O values and the overall δ18O value of the cellulose. We apply the method to determine the degree to which oxygen atoms at each position undergo isotopic exchange with water during heterotrophic cellulose synthesis, such as occurs in the cambium of trees. To do this we extract α-cellulose from wheat seedlings germinated in the dark in aqueous media of differing oxygen isotope ratios. Results indicate that oxygen atoms at positions 5 and 6 (O-5 and O-6 respectively) undergo around 80% exchange with medium water, O-3 undergoes around 50% exchange, and O-2 and O-4 do not undergo isotopic exchange. The results have important implications for extracting palaeoclimatic records from oxygen isotope time series obtained from tree ring cellulose. As O-5 and O-6 undergo significant exchange with medium water during heterotrophic cellulose synthesis, oxygen isotopes at these positions in tree ring cellulose should carry a predominantly trunk (source) water signal. On the other hand, O-2 and O-4 should retain the isotopic signature of leaf water in tree ring cellulose. Our method therefore potentially enables the separate reconstruction of past temperature and humidity data from oxygen isotope ratios of tree ring cellulose - something that has hitherto not been possible. The measured degrees of isotopic exchange are to some extent unexpected and cannot be fully explained using current biochemical mechanisms, suggesting that knowledge of these processes is incomplete.

  11. Measurement of human growth hormone in urine: development and validation of a sensitive and specific assay.

    PubMed

    Hourd, P; Edwards, R

    1989-04-01

    A specific solid-phase immunoradiometric assay (IRMA), optimized for maximum sensitivity, has been developed for measurement of human GH (hGH) in urine. The sensitivity varied with sample size, giving a range of 0.001 to 0.003 mU/l for a sample volume of 2 ml. Recovery and dilution experiments, together with chromatography of urine samples, indicate that the method is specific for hGH. Added exogenous hGH was measured with a mean recovery of 101 +/- 10% (S.D.) for 1 ml samples and 87 +/- 8% for 2 ml samples. Measurements of samples diluted at 1:2 and 1:4 gave values of 97.4 and 96.6% respectively of those expected. Cross-reactions of human placental lactogen and prolactin were less than 0.008 and 0.04% respectively on a mol/mol basis. The assay was insensitive to the presence of NaCl (50-500 mmol/l), urea (50-1000 mmol/l), creatinine (1-20 mmol/l), Ca2+ ions (1-20 mmol/l), SO4(2-) ions (1-1000 mmol/l), Mg2+ ions (0.05-50 mmol/l), 0.5-5% (w/v) glucose and a pH range of 6-9. Chromatography of unextracted samples showed that the immunoreactive material in urine eluted in a single homogenous peak with a similar position to monomeric pituitary hGH (22 kDa). Administered hGH (0.002%) was recovered in urine collected over a 2-h period following an intravenous injection. The urine output of hGH showed a good correlation with serum hGH in 18 patients following routine insulin tolerance tests and in 25 patients following an oral glucose tolerance test.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2715756

  12. Activation mechanism of thiol protease precursor from broiler chicken specific Staphylococcus aureus strain CH-91.

    PubMed

    Wladyka, Benedykt; Dubin, Grzegorz; Dubin, Adam

    2011-01-10

    Staphylococcus aureus strain CH-91 isolated from chicken dermatitis lesions produces large quantities of thiol protease implicated in disease formation. Observed overproduction requires efficient activation of the protease precursor which mechanism is studied here in detail. Wild type and mutant precursor forms are expressed in E. coli to test different hypotheses on the activation process. It is demonstrated that wild type precursor undergoes rapid autocatalytic processing whereas proteolytically inactive catalytic triad cysteine mutant (C(249)A) of the precursor is stable, but can be processed by minute quantities of active protease. It is concluded that limited intramolecular proteolysis is mainly responsible for efficient activation but, a positive feedback loop also contributes to the process. Both activation pathways allow efficient production of mature extracellular thiol protease, a putative virulence factor specific for avian strains of S. aureus. PMID:20598816

  13. Specific pollen allergen activates eosinophils of the patient with chronic allergic contact urticaria.

    PubMed

    Panaszek, B; Małolepszy, J; Kuryszko, J; Litwa, M

    1994-01-01

    The aim of the study was to evaluate the activation of eosinophils in an unique case of a young man with atopy manifested as chronic pollen contact urticaria. In order to reveal the role of eosinophils in that case, the study was performed by means of monoclonal antibodies EG2 and chemiluminescence. In addition, comparative electron microscopic study of peripheral blood and skin infiltrating eosinophils were performed for which the name ultrastructural morphometric analysis of intracytoplasmic eosinophil granules has been proposed. The results indicated, that 40% of peripheral blood eosinophils were activated spontaneously and they were more active than those in skin infiltrates. Specific pollen allergen caused activation of 100% of peripheral blood eosinophils. The study suggests presence of a systemic pattern of eosinophil activation in atopy. PMID:7487362

  14. Microbial metabolic activity in soil as measured by dehydrogenase determinations

    NASA Technical Reports Server (NTRS)

    Casida, L. E., Jr.

    1977-01-01

    The dehydrogenase technique for measuring the metabolic activity of microorganisms in soil was modified to use a 6-h, 37 C incubation with either glucose or yeast extract as the electron-donating substrate. The rate of formazan production remained constant during this time interval, and cellular multiplication apparently did not occur. The technique was used to follow changes in the overall metabolic activities of microorganisms in soil undergoing incubation with a limiting concentration of added nutrient. The sequence of events was similar to that obtained by using the Warburg respirometer to measure O2 consumption. However, the major peaks of activity occurred earlier with the respirometer. This possibly is due to the lack of atmospheric CO2 during the O2 consumption measurements.

  15. An Alternative Measure of Solar Activity from Detailed Sunspot Datasets

    NASA Astrophysics Data System (ADS)

    Muraközy, J.; Baranyi, T.; Ludmány, A.

    2016-05-01

    The sunspot number is analyzed by using detailed sunspot data, including aspects of observability, sunspot sizes, and proper identification of sunspot groups as discrete entities of solar activity. The tests show that in addition to the subjective factors there are also objective causes of the ambiguities in the series of sunspot numbers. To introduce an alternative solar-activity measure, the physical meaning of the sunspot number has to be reconsidered. It contains two components whose numbers are governed by different physical mechanisms and this is one source of the ambiguity. This article suggests an activity index, which is the amount of emerged magnetic flux. The only long-term proxy measure is the detailed sunspot-area dataset with proper calibration to the magnetic flux. The Debrecen sunspot databases provide an appropriate source for the establishment of the suggested activity index.

  16. Reactor Physics Measurements and Benchmark Specifications for Oak Ridge Highly Enriched Uranium Sphere (ORSphere)

    DOE PAGESBeta

    Marshall, Margaret A.

    2014-11-04

    In the early 1970s Dr. John T. Mihalczo (team leader), J.J. Lynn, and J.R. Taylor performed experiments at the Oak Ridge Critical Experiments Facility (ORCEF) with highly enriched uranium (HEU) metal (called Oak Ridge Alloy or ORALLOY) in an effort to recreate GODIVA I results with greater accuracy than those performed at Los Alamos National Laboratory in the 1950s. The purpose of the Oak Ridge ORALLOY Sphere (ORSphere) experiments was to estimate the unreflected and unmoderated critical mass of an idealized sphere of uranium metal corrected to a density, purity, and enrichment such that it could be compared with themore » GODIVA I experiments. Additionally, various material reactivity worths, the surface material worth coefficient, the delayed neutron fraction, the prompt neutron decay constant, relative fission density, and relative neutron importance were all measured. The critical assembly, material reactivity worths, the surface material worth coefficient, and the delayed neutron fraction were all evaluated as benchmark experiment measurements. The reactor physics measurements are the focus of this paper; although for clarity the critical assembly benchmark specifications are briefly discussed.« less

  17. Reactor Physics Measurements and Benchmark Specifications for Oak Ridge Highly Enriched Uranium Sphere (ORSphere)

    SciTech Connect

    Marshall, Margaret A.

    2014-11-04

    In the early 1970s Dr. John T. Mihalczo (team leader), J.J. Lynn, and J.R. Taylor performed experiments at the Oak Ridge Critical Experiments Facility (ORCEF) with highly enriched uranium (HEU) metal (called Oak Ridge Alloy or ORALLOY) in an effort to recreate GODIVA I results with greater accuracy than those performed at Los Alamos National Laboratory in the 1950s. The purpose of the Oak Ridge ORALLOY Sphere (ORSphere) experiments was to estimate the unreflected and unmoderated critical mass of an idealized sphere of uranium metal corrected to a density, purity, and enrichment such that it could be compared with the GODIVA I experiments. Additionally, various material reactivity worths, the surface material worth coefficient, the delayed neutron fraction, the prompt neutron decay constant, relative fission density, and relative neutron importance were all measured. The critical assembly, material reactivity worths, the surface material worth coefficient, and the delayed neutron fraction were all evaluated as benchmark experiment measurements. The reactor physics measurements are the focus of this paper; although for clarity the critical assembly benchmark specifications are briefly discussed.

  18. Measuring the relative extent of pulmonary infiltrates by hierarchical classification of patient-specific image features

    NASA Astrophysics Data System (ADS)

    Tsevas, S.; Iakovidis, D. K.

    2011-11-01

    Pulmonary infiltrates are common radiological findings indicating the filling of airspaces with fluid, inflammatory exudates, or cells. They are most common in cases of pneumonia, acute respiratory syndrome, atelectasis, pulmonary oedema and haemorrhage, whereas their extent is usually correlated with the extent or the severity of the underlying disease. In this paper we propose a novel pattern recognition framework for the measurement of the extent of pulmonary infiltrates in routine chest radiographs. The proposed framework follows a hierarchical approach to the assessment of image content. It includes the following: (a) sampling of the lung fields; (b) extraction of patient-specific grey-level histogram signatures from each sample; (c) classification of the extracted signatures into classes representing normal lung parenchyma and pulmonary infiltrates; (d) the samples for which the probability of belonging to one of the two classes does not reach an acceptable level are rejected and classified according to their textural content; (e) merging of the classification results of the two classification stages. The proposed framework has been evaluated on real radiographic images with pulmonary infiltrates caused by bacterial infections. The results show that accurate measurements of the infiltration areas can be obtained with respect to each lung field area. The average measurement error rate on the considered dataset reached 9.7% ± 1.0%.

  19. A Prototype RF Dosimeter for Independent Measurement of the Average Specific Absorption Rate (SAR) During MRI

    PubMed Central

    Stralka, John P; Bottomley, Paul A

    2008-01-01

    Purpose To develop a scanner-independent dosimeter for measuring the average radio frequency (RF) power deposition and specific absorption rates (SAR) for human MRI exposure. Materials and Methods A prototype dosimeter has a transducer with orthogonal conducting loops surrounding a small signal-generating MRI sample. The loops contain resistors whose values are adjusted to load the scanner’s MRI coils equivalent to an average head or body during MRI. The scanner adjusts its power output to normal levels during setup, using the MRI sample. Following calibration, the total power and average SAR deposited in the transducer are measured from the root-mean-square (rms) power induced in the transducer during MRI. Results A 1.5 Tesla head transducer was adjusted to elicit the same load as the average of nine adult volunteers. Once adjusted, the transducer loads other head coils the same as the head does. The dosimeter is calibrated at up to 20 W total deposited power and 4.5 W/kg SAR in the average head, with about 5% accuracy. Conclusion This dosimeter provides a simple portable means of measuring the power deposited in a body-equivalent sample load, independent of the scanner. Further work will develop SAR dosimetry for the torso and for higher fields. PMID:17969145

  20. Photospheric Magnetic Diffusion by Measuring Moments of Active Regions

    NASA Astrophysics Data System (ADS)

    Engell, Alexander; Longcope, D.

    2013-07-01

    Photospheric magnetic surface diffusion is an important constraint for the solar dynamo. The HMI Active Region Patches (HARPs) program automatically identify all magnetic regions above a certain flux. In our study we measure the moments of ARs that are no longer actively emerging and can thereby give us good statistical constraints on photospheric diffusion. We also present the diffusion properties as a function of latitude, flux density, and single polarity (leading or following) within each HARP.

  1. Autoantigen-Specific B Cell Activation in FAS-Deficient Rheumatoid Factor Immunoglobulin Transgenic Mice

    PubMed Central

    Wang, Haowei; Shlomchik, Mark J.

    1999-01-01

    In systemic autoimmune disease, self-tolerance fails, leading to autoantibody production. A central issue in immunology is to understand the origins of activated self-reactive B cells. We have used immunoglobulin (Ig) transgenic mice to investigate the regulation of autoreactive B cells with specificity for self-IgG2a (the rheumatoid factor [RF] specificity) to understand how normal mice regulate RF autoantibodies and how this fails in autoimmune mice. We previously showed that normal mice do not tolerize the AM14 RF clone, nor do they appear to activate it. Here we show that in Fas-deficient autoimmune mice, the picture is quite different. RF B cells are activated to divide and secrete, but only when the autoantigen is present. Thus, B cells that are ignored rather than anergized in normal mice can be stimulated to produce autoantibody in Fas-deficient mice. This demonstrates a novel developmental step at which intact Fas–Fas ligand signaling is required to regulate B cells in order to prevent autoimmunity. These data also establish the relevance of ignorant self-specific B cells to autoantibody production in disease and prove that in the case of the RF specificity, the nominal autoantigen IgG2a is the driving autoantigen in vivo. PMID:10477549

  2. Successful Remembering Elicits Event-Specific Activity Patterns in Lateral Parietal Cortex

    PubMed Central

    Chun, Marvin M.

    2014-01-01

    Remembering a past event involves reactivation of content-specific patterns of neural activity in high-level perceptual regions (e.g., ventral temporal cortex, VTC). In contrast, the subjective experience of vivid remembering is typically associated with increased activity in lateral parietal cortex (LPC)—“retrieval success effects” that are thought to generalize across content types. However, the functional significance of LPC activation during memory retrieval remains a subject of active debate. In particular, theories are divided with respect to whether LPC actively represents retrieved content or if LPC activity only scales with content reactivation elsewhere (e.g., VTC). Here, we report a human fMRI study of visual memory recall (faces vs scenes) in which complementary forms of multivoxel pattern analysis were used to test for and compare content reactivation within LPC and VTC. During recall of visual images, we observed robust reactivation of broad category information (face vs scene) in both VTC and LPC. Moreover, recall-related activity patterns in LPC, but not VTC, differentiated between individual events. Importantly, these content effects were particularly evident in areas of LPC (namely, angular gyrus) in which activity scaled with subjective reports of recall vividness. These findings provide striking evidence that LPC not only signals that memories have been successfully recalled, but actively represents what is being remembered. PMID:24899726

  3. Identification of novel target genes specifically activated by deregulated E2F in human normal fibroblasts.

    PubMed

    Kitamura, Hodaka; Ozono, Eiko; Iwanaga, Ritsuko; Bradford, Andrew P; Okuno, Junko; Shimizu, Emi; Kurayoshi, Kenta; Kugawa, Kazuyuki; Toh, Hiroyuki; Ohtani, Kiyoshi

    2015-09-01

    The transcription factor E2F is the principal target of the tumor suppressor pRB. E2F plays crucial roles not only in cell proliferation by activating growth-related genes but also in tumor suppression by activating pro-apoptotic and growth-suppressive genes. We previously reported that, in human normal fibroblasts, the tumor suppressor genes ARF, p27(Kip1) and TAp73 are activated by deregulated E2F activity induced by forced inactivation of pRB, but not by physiological E2F activity induced by growth stimulation. In contrast, growth-related E2F targets are activated by both E2F activities, underscoring the roles of deregulated E2F in tumor suppression in the context of dysfunctional pRB. In this study, to further understand the roles of deregulated E2F, we explored new targets that are specifically activated by deregulated E2F using DNA microarray. The analysis identified nine novel targets (BIM, RASSF1, PPP1R13B, JMY, MOAP1, RBM38, ABTB1, RBBP4 and RBBP7), many of which are involved in the p53 and RB tumor suppressor pathways. Among these genes, the BIM gene was shown to be activated via atypical E2F-responsive promoter elements and to contribute to E2F1-mediated apoptosis. Our results underscore crucial roles of deregulated E2F in growth suppression to counteract loss of pRB function. PMID:26201719

  4. Phase advancement and nucleus-specific timing of thalamocortical activity during slow cortical oscillation

    PubMed Central

    Slézia, Andrea; Hangya, Balázs; Ulbert, István; Acsády, László

    2011-01-01

    The exact timing of cortical afferent activity is instrumental for the correct coding and retrieval of internal and external stimuli. Thalamocortical inputs represent the most significant subcortical pathway to the cortex, but the precise timing and temporal variability of thalamocortical activity is not known. To examine this question, we studied the phase of thalamic action potentials relative to cortical oscillations and established correlations among phase, the nuclear location of the thalamocortical neurons and the frequency of cortical activity. The phase of thalamic action potentials depended on the exact frequency of the slow cortical oscillation both on long (minutes) and short (single wave) time scales. Faster waves were accompanied by phase advancement in both cases. Thalamocortical neurons located in different nuclei fired at significantly different phases of the slow waves but were active at similar phase of spindle oscillations. Different thalamic nuclei displayed distinct burst patterns. Bursts with higher number of action potentials displayed progressive phase advancement in a nucleus-specific manner. Thalamic neurons located along nuclear borders were characterized by mixed burst and phase properties. Our data demonstrate that the temporal relationship between cortical and thalamic activity is not fixed but displays dynamic changes during oscillatory activity. The timing depends on the precise location and exact activity of thalamocortical cells and the ongoing cortical network pattern. This variability of thalamic output and its coupling to cortical activity can enable thalamocortical neurons to actively participate in the coding and retrieval of complex cortical signals. PMID:21228169

  5. Only Specific Tobacco (Nicotiana tabacum) Chitinases and [beta]-1,3-Glucanases Exhibit Antifungal Activity.

    PubMed Central

    Sela-Buurlage, M. B.; Ponstein, A. S.; Bres-Vloemans, S. A.; Melchers, L. S.; Van Den Elzen, PJM.; Cornelissen, BJC.

    1993-01-01

    Different isoforms of chitinases and [beta]-1,3-glucanases of tobacco (Nicotiana tabacum cv Samsun NN) were tested for their antifungal activities. The class I, vacuolar chitinase and [beta]-1,3-glucanase isoforms were the most active against Fusarium solani germlings, resulting in lysis of the hyphal tips and in growth inhibition. In additon, we observed that the class I chitinase and [beta]-1,3-glucanase acted synergistically. The class II isoforms of the two hydrolases exhibited no antifungal activity. However, the class II chitinases showed limited growth inhibitory activity in combination with higher amounts of class I [beta]-1,3-glucanase. The class II [beta]-1,3-glucanases showed no inhibitory activity in any combination. In transgenic tobacco plants producing modified forms of either a class I chitinase or a class I [beta]-1,3-glucanase, or both, these proteins were targeted extracellularly. Both modified proteins lack their C-terminal propeptide, which functions as a vacuolar targeting signal. Extracellular targeting had no effect on the specific activities of the chitinase and [beta]-1,3-glucanase enzymes. Furthermore, the extracellular washing fluid (EF) from leaves of transgenic plants expressing either of the secreted class I enzymes exhibited antifungal activity on F. solani germlings in vitro comparable to that of the purified vacuolar class I proteins. Mixing EF fractions from these plants revealed synergism in inhibitory activity against F. solani; the mixed fractions exhibited inhibitory activity similar to that of EF from plants expressing both secreted enzymes. PMID:12231736

  6. Spectrometric measurements of radioisotope activity in the thyroid

    NASA Astrophysics Data System (ADS)

    Osko, Jakub; Golnik, Natalia

    2008-01-01

    The results of measurements of iodine 131I and technetium 99mTc uptake in human thyroid, performed with scintillation or semiconductor detectors can exhibit a considerable uncertainty due to the differences in the thyroid position in the patient's neck. Basic physical laws of radiation attenuation and scattering show that the final shape of the registered spectrum should depends on the thyroid position in the neck and on the thickness of the tissue between the thyroid and the detector. The use of the spectrometric measuring method is proposed in this work for determination of the iodine gathering effective depth. The performed studies showed that the measurements results can be used for improving the accuracy of the iodine 131I activity in thyroid measurements and for selection of the group of patients for whom the anatomical position of the thyroid or the spatial distribution of the iodine gathering is much different than the standard one, assumed during the calibration of the counters. The results of the measurements were in agreement with Monte-Carlo calculations of the detector response. The method was used in routine monitoring of occupationally exposed persons, using the thyroid counter. A group of six persons with measurable internal contamination was selected and the measurements were performed on consecutive days, so the results could be registered at decreasing iodine activities in the thyroid. Larger series of measurements were performed at Brodno Regional Hospital in Warsaw, for a group of 95 patients after diagnostic administration of iodine 131I.

  7. The influence of thoron on instruments measuring radon activity concentration.

    PubMed

    Michielsen, N; Bondiguel, S

    2015-11-01

    Thoron, the isotope 220 of radon, is a radionuclide whose concentration may influence the measurement of the activity concentration of (222)Rn in the air. If in the case of continuous and active sampling measuring instruments, using a pump for example, the influence of thoron on radon measurement is obvious and is taken into account in the apparatus, it is often assumed that in the case of a passive sampling, by diffusion through a filter for example, this thoron influence is negligible. This is due to the very short radioactive half-life of thoron, 55.6 s (3.82 d for (222)Rn), and the assumption that the diffusion time of thoron in the detection chamber is long enough beside that of the thoron half-life. The objective of this study is to check whether this assumption is true or not for different kinds of commercial electronic apparatus used to measure radon activity concentration from soil to dwellings. First of all, the devices were calibrated in activity concentration of radon, and then they were exposed to a controlled thoron atmosphere. The experiments concerning the thoron aimed to investigate the sensitivity to thoron in the radon measuring mode of the apparatus. Results of these experiments show that all devices have a very quick answer to thoron atmosphere, even though the sensitivities vary from one instrument to another. Results clearly show that this influence on radon measurement due to the thoron is observed also after the exposition because of the decay of (212)Pb and its progenies. In conclusion, the sensitivity to thoron in the radon measuring mode depends strongly on the type of instruments. The results of the present investigation show that for some apparatus, the influence of thoron cannot be disregarded especially when measuring radon in soil. PMID:25953787

  8. Site-specific phosphorylation and microtubule dynamics control Pyrin inflammasome activation.

    PubMed

    Gao, Wenqing; Yang, Jieling; Liu, Wang; Wang, Yupeng; Shao, Feng

    2016-08-16

    Pyrin, encoded by the MEFV gene, is best known for its gain-of-function mutations causing familial Mediterranean fever (FMF), an autoinflammatory disease. Pyrin forms a caspase-1-activating inflammasome in response to inactivating modifications of Rho GTPases by various bacterial toxins or effectors. Pyrin-mediated innate immunity is unique in that it senses bacterial virulence rather than microbial molecules, but its mechanism of activation is unknown. Here we show that Pyrin was phosphorylated in bone marrow-derived macrophages and dendritic cells. We identified Ser-205 and Ser-241 in mouse Pyrin whose phosphorylation resulted in inhibitory binding by cellular 14-3-3 proteins. The two serines underwent dephosphorylation upon toxin stimulation or bacterial infection, triggering 14-3-3 dissociation, which correlated with Pyrin inflammasome activation. We developed antibodies specific for phosphorylated Ser-205 and Ser-241, which confirmed the stimuli-induced dephosphorylation of endogenous Pyrin. Mutational analyses indicated that both phosphorylation and signal-induced dephosphorylation of Ser-205/241 are important for Pyrin activation. Moreover, microtubule drugs, including colchicine, commonly used to treat FMF, effectively blocked activation of the Pyrin inflammasome. These drugs did not affect Pyrin dephosphorylation and 14-3-3 dissociation but inhibited Pyrin-mediated apoptosis-associated Speck-like protein containing CARD (ASC) aggregation. Our study reveals that site-specific (de)phosphorylation and microtubule dynamics critically control Pyrin inflammasome activation, illustrating a fine and complex mechanism in cytosolic immunity. PMID:27482109

  9. 2,4-Dichlorophenol hydroxylase for chlorophenol removal: Substrate specificity and catalytic activity.

    PubMed

    Ren, Hejun; Li, Qingchao; Zhan, Yang; Fang, Xuexun; Yu, Dahai

    2016-01-01

    Chlorophenols (CPs) are common environmental pollutants. As such, different treatments have been assessed to facilitate their removal. In this study, 2,4-dichlorophenol (2,4-DCP) hydroxylase was used to systematically investigate the activity and removal ability of 19CP congeners at 25 and 0 °C. Results demonstrated that 2,4-DCP hydroxylase exhibited a broad substrate specificity to CPs. The activities of 2,4-DCP hydroxylase against specific CP congeners, including 3-CP, 2,3,6-trichlorophenol, 2-CP, and 2,3-DCP, were higher than those against 2,4-DCP, which is the preferred substrate of previously reported 2,4-DCP hydroxylase. To verify whether cofactors are necessary to promote hydroxylase activity against CP congeners, we added FAD and found that the added FAD induced a 1.33-fold to 5.13-fold significant increase in hydroxylase activity against different CP congeners. The metabolic pathways of the CP degradation in the enzymatic hydroxylation step were preliminarily proposed on the basis of the analyses of the enzymatic activities against 19CP congeners. We found that the high activity and removal rate of 2,4-DCP hydroxylase against CPs at 0 °C enhance the low-temperature-adaptability of this enzyme to the CP congeners; as such, the proposed removal process may be applied to biochemical, bioremediation, and industrial processes, particularly in cold environments. PMID:26672451

  10. Kinetic isotope effects for RNA cleavage by 2'-O- transphosphorylation: Nucleophilic activation by specific base

    PubMed Central

    Harris, Michael E; Dai, Qing; Gu, Hong; Kellerman, Dan; Piccirilli, Joseph A; Anderson, Vernon E

    2010-01-01

    To better understand the interactions between catalysts and transition states during RNA strand cleavage, primary 18O kinetic isotope effects and solvent D2O isotope effects were measured to probe the mechanism of base-catalyzed 2'-O-transphosphorylation of the RNA dinucleotide 5'-UpG-3'. The observed 18O KIEs for the nucleophilic 2'-O and in the 5'-O leaving group at pH 14 are both large relative to reactions of phosphodiesters with good leaving groups, indicating that the reaction catalyzed by hydroxide has a transition state (TS) with advanced phosphorus-oxygen bond fission to the leaving group (18kLG = 1.034 ± 0.004) and phosphorous-nucleophile bond formation (18kNUC = 0.984 ± 0.004). A breakpoint in the pH dependence of the 2'-O-transphosphorylation rate to a pH independent phase above pH 13 has been attributed to the pKa of the 2'-OH nucleophile. A smaller nucleophile KIE is observed at pH 12 (18kNUC = 0.995 ± 0.004) that is interpreted as the combined effect of the equilibrium isotope effect (~1.02) on deprotonation of the 2′-hydroxyl nucleophile and the intrinsic KIE on the nucleophilic addition step (ca. 0.981). An alternative mechanism in which the hydroxide ion acts as a general base is considered unlikely given the lack of a solvent deuterium isotope effect above the breakpoint in the pH versus rate profile. These results represent the first direct analysis of the transition state for RNA strand cleavage. The primary 18O KIE results and the lack of a kinetic solvent deuterium isotope effect together provide strong evidence for a late transition state and 2'-O nucleophile activation by specific base catalysis. PMID:20669950

  11. Prefusion F–specific antibodies determine the magnitude of RSV neutralizing activity in human sera

    PubMed Central

    Ngwuta, Joan O.; Chen, Man; Modjarrad, Kayvon; Joyce, M. Gordon; Kanekiyo, Masaru; Kumar, Azad; Yassine, Hadi M.; Moin, Syed M.; Killikelly, April M.; Chuang, Gwo-Yu; Druz, Aliaksandr; Georgiev, Ivelin S.; Rundlet, Emily J.; Sastry, Mallika; Stewart-Jones, Guillaume B. E.; Yang, Yongping; Zhang, Baoshan; Nason, Martha C.; Capella, Cristina; Peeples, Mark E.; Ledgerwood, Julie E.; McLellan, Jason S.; Kwong, Peter D.; Graham, Barney S.

    2015-01-01

    Respiratory syncytial virus (RSV) is estimated to claim more lives among infants <1 year old than any other single pathogen, except malaria, and poses a substantial global health burden. Viral entry is mediated by a type I fusion glycoprotein (F) that transitions from a metastable prefusion (pre-F) to a stable postfusion (post-F) trimer. A highly neutralization-sensitive epitope, antigenic site Ø, is found only on pre-F. We determined what fraction of neutralizing (NT) activity in human sera is dependent on antibodies specific for antigenic site Ø or other antigenic sites on F in healthy subjects from ages 7 to 93 years. Adsorption of individual sera with stabilized pre-F protein removed >90% of NT activity and depleted binding antibodies to both F conformations. In contrast, adsorption with post-F removed ~30% of NT activity, and binding antibodies to pre-F were retained. These findings were consistent across all age groups. Protein competition neutralization assays with pre-F mutants in which sites Ø or II were altered to knock out binding of antibodies to the corresponding sites showed that these sites accounted for ~35 and <10% of NT activity, respectively. Binding competition assays with monoclonal antibodies (mAbs) indicated that the amount of site Ø–specific antibodies correlated with NT activity, whereas the magnitude of binding competed by site II mAbs did not correlate with neutralization. Our results indicate that RSV NT activity in human sera is primarily derived from pre-F–specific antibodies, and therefore, inducing or boosting NT activity by vaccination will be facilitated by using pre-F antigens that preserve site Ø. PMID:26468324

  12. Active-site Arg --> Lys substitutions alter reaction and substrate specificity of aspartate aminotransferase.

    PubMed

    Vacca, R A; Giannattasio, S; Graber, R; Sandmeier, E; Marra, E; Christen, P

    1997-08-29

    Arg386 and Arg292 of aspartate aminotransferase bind the alpha and the distal carboxylate group, respectively, of dicarboxylic substrates. Their substitution with lysine residues markedly decreased aminotransferase activity. The kcat values with L-aspartate and 2-oxoglutarate as substrates under steady-state conditions at 25 degrees C were 0.5, 2.0, and 0.03 s-1 for the R292K, R386K, and R292K/R386K mutations, respectively, kcat of the wild-type enzyme being 220 s-1. Longer dicarboxylic substrates did not compensate for the shorter side chain of the lysine residues. Consistent with the different roles of Arg292 and Arg386 in substrate binding, the effects of their substitution on the activity toward long chain monocarboxylic (norleucine/2-oxocaproic acid) and aromatic substrates diverged. Whereas the R292K mutation did not impair the aminotransferase activity toward these substrates, the effect of the R386K substitution was similar to that on the activity toward dicarboxylic substrates. All three mutant enzymes catalyzed as side reactions the beta-decarboxylation of L-aspartate and the racemization of amino acids at faster rates than the wild-type enzyme. The changes in reaction specificity were most pronounced in aspartate aminotransferase R292K, which decarboxylated L-aspartate to L-alanine 15 times faster (kcat = 0.002 s-1) than the wild-type enzyme. The rates of racemization of L-aspartate, L-glutamate, and L-alanine were 3, 5, and 2 times, respectively, faster than with the wild-type enzyme. Thus, Arg --> Lys substitutions in the active site of aspartate aminotransferase decrease aminotransferase activity but increase other pyridoxal 5'-phosphate-dependent catalytic activities. Apparently, the reaction specificity of pyridoxal 5'-phosphate-dependent enzymes is not only achieved by accelerating the specific reaction but also by preventing potential side reactions of the coenzyme substrate adduct. PMID:9268327

  13. Enrichment and isolation of a ruminal bacterium with a very high specific activity of ammonia production.

    PubMed Central

    Russell, J B; Strobel, H J; Chen, G J

    1988-01-01

    When mixed ruminal bacteria were inoculated into semicontinuous cultures (25% transfer every other day) containing lactate, dulcitol, pectin, or xylose and Trypticase (1 g/liter) as the sole nitrogen source, the specific activity of ammonia production increased. The greatest enrichment was observed with lactate and xylose, and in these cases the specific rate of ammonia production was eightfold higher than that of the ruminal fluid control (approximately 35 nmol of ammonia per mg of protein per min). Isolates with different morphologies were obtained from each of the enrichments, but in no case did the specific activity of any isolate exceed that of the mixed ruminal bacteria. If Trypticase (15 g/liter) was used as the only energy and nitrogen source, there was an even greater increase in ammonia production, and two monensin-sensitive bacteria, a Peptostreptococcus species and a Clostridium species, were obtained. The Peptostreptococcus species was unable to grow on any of 25 carbohydrate or carbohydrate derivatives tested; but the Clostridium species was able to use glucose, maltose, fructose, cellobiose, trehalose, sorbitol, and salicin as energy sources. Neither organism was able to grow in the absence of an amino acid source, but growth rates on Trypticase were greater than 0.35/h. The specific activities of ammonia production were 346 and 427 nmol/mg of protein per min for strains of Peptostreptococcus and Clostridium, respectively. Megasphaera elsdenii and Bacteroides ruminicola, previously isolated ruminal ammonia producers, had specific activities of only 11 and 19 nmol of ammonia per mg of protein per min, respectively. The most probable number of Clostridium species in ruminal fluid was less than 10(3)/ml, but the Peptostreptococcus species was present at 10(8)/ml.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3377500

  14. Measuring emotion in advertising research: prefrontal brain activity.

    PubMed

    Silberstein, Richard B; Nield, Geoffrey E

    2012-01-01

    With the current interest in the role of emotion in advertising and advertising research, there has been an increasing interest in the use of various brain activity measures to access nonverbal emotional responses. One such approach relies on measuring the difference between left and right hemisphere prefrontal cortical activity to assess like and dislike. This approach is based on electroencephalography (EEG) and neuroimaging work, suggesting that the approach/withdrawal (frequently but not always associated with like/dislike) dimension of emotion is indicated by the balance of activity between the left and right prefrontal cortex. Much of this work was initiated by Richard Davidson in the early 1990s. An early study by Davidson et al. measured brain electrical activity to assess patterns of activation during the experience of happiness and disgust. The authors reported that disgust was found to be associated with increased right-sided activation in the frontal and anterior temporal regions compared with happiness. In contrast, happiness was found to be accompanied by left-sided activation in the anterior temporal region compared with disgust. Early reports suggested that frontal laterality indexes motivational valence with positive emotions (happy, like) associated with left greater than the right frontal activity and vice versa. Although these findings appear to be consistent with personality traits (e.g., optimism pessimism), state changes in frontal laterality appears to index approach withdraw rather than emotional valence. Interestingly, the behavioral and motivational correlates of prefrontal asymmetric activity are not restricted to humans or even primates but have been observed in numerous species such as birds and fish (see [4]). Henceforth, we use the term motivational valence (MV) rather than the more cumbersome term approach withdraw. PMID:22678836

  15. Novel method demonstrates differential ligand activation and phosphatase-mediated deactivation of insulin receptor tyrosine-specific phosphorylation.

    PubMed

    Cieniewicz, Anne M; Cooper, Philip R; McGehee, Jennifer; Lingham, Russell B; Kihm, Anthony J

    2016-08-01

    Insulin receptor signaling is a complex cascade leading to a multitude of intracellular functional responses. Three natural ligands, insulin, IGF1 and IGF2, are each capable of binding with different affinities to the insulin receptor, and result in variable biological responses. However, it is likely these affinity differences alone cannot completely explain the myriad of diverse cellular outcomes. Ligand binding initiates activation of a signaling cascade resulting in phosphorylation of the IR itself and other intracellular proteins. The direct catalytic activity along with the temporally coordinated assembly of signaling proteins is critical for insulin receptor signaling. We hypothesized that determining differential phosphorylation among individual tyrosine sites activated by ligand binding or dephosphorylation by phosphatases could provide valuable insight into insulin receptor signaling. Here, we present a sensitive, novel immunoassay adapted from Meso Scale Discovery technology to quantitatively measure changes in site-specific phosphorylation levels on endogenous insulin receptors from HuH7 cells. We identified insulin receptor phosphorylation patterns generated upon differential ligand activation and phosphatase-mediated deactivation. The data demonstrate that insulin, IGF1 and IGF2 elicit different insulin receptor phosphorylation kinetics and potencies that translate to downstream signaling. Furthermore, we show that insulin receptor deactivation, regulated by tyrosine phosphatases, occurs distinctively across specific tyrosine residues. In summary, we present a novel, quantitative and high-throughput assay that has uncovered differential ligand activation and site-specific deactivation of the insulin receptor. These results may help elucidate some of the insulin signaling mechanisms, discriminate ligand activity and contribute to a better understanding of insulin receptor signaling. We propose this methodology as a powerful approach to characterize

  16. Pancreas-specific lipase concentrations and amylase and lipase activities in the peritoneal fluid of dogs with suspected pancreatitis.

    PubMed

    Chartier, Marie A; Hill, Steve L; Sunico, Sarena; Suchodolski, Jan S; Robertson, Jane E; Steiner, Joerg M

    2014-09-01

    Diagnosing acute pancreatitis in the dog can be challenging. The aim of this study was to determine the concentrations of pancreas-specific lipase immunoreactivity (cPLI), and the activities of amylase and lipase, in the peritoneal fluid from a population of dogs diagnosed with acute pancreatitis based on clinical signs, ultrasonographic findings and serum cPLI concentrations. In a prospective study, cPLI concentrations, and amylase and lipase activities, were measured in the peritoneal fluid of 14 dogs with pancreatitis and 19 dogs with non-pancreatic disease. The sensitivity and specificity of peritoneal fluid cPLI concentration (cut-off value 500 µg/L) were 100.0% (95% confidence interval, CI, 80.7-100.0%) and 94.7% (95% CI 76.7-99.7%), respectively. The sensitivity and specificity of peritoneal fluid amylase (cut-off value 1050 U/L) and lipase activities (cut-off value 500 U/L) were 71.4% (95% CI 44.5-90.2%) and 84.2% (95% CI 62.8-95.8%) for amylase activity, and 92.9% (95% CI 69.5-99.6%) and 94.7% (95% CI 76.7-99.7%) for lipase activity, respectively. In conclusion, peritoneal fluid cPLI concentration was highly sensitive as a complementary diagnostic tool in a group of dogs with suspected acute pancreatitis. Peritoneal fluid lipase activity was not as sensitive as cPLI concentration, but may also support a diagnosis of acute pancreatitis in dogs. PMID:25106805

  17. Production of 191Pt radiotracer with high specific activity for the development of preconcentration procedures

    NASA Astrophysics Data System (ADS)

    Parent, M.; Strijckmans, K.; Cornelis, R.; Dewaele, J.; Dams, R.

    1994-04-01

    A radiotracer of Pt with suitable nuclear characteristics and high specific activity (i.e. activity to mass ratio) is a powerful tool when developing preconcentration methods for the determination of base-line levels of Pt in e.g. environmental and biological samples. Two methods were developed for the production of 191Pt with high specific activity and radionuclidic purity: (1) via the 190Pt(n, γ) 191Pt reaction by neutron irradiation of enriched Pt in a nuclear reactor at high neutron fluence rate and (2) via the 191Ir(p, n) 191Pt reaction by proton irradiation of natural Ir with a cyclotron, at an experimentally optimized proton energy. For the latter method it was necessary to separate Pt from the Ir matrix. For that reason either liquid-liquid extraction with dithizone or adsorption chromatography were used. The yields, the specific activities and the radionuclidic purities were experimentally determined as a function of the proton energy and compared to the former method. The half-life of 191Pt was accurately determined to be 2.802 ± 0.025 d.

  18. Functional adaptation of cortical interneurons to attenuated activity is subtype-specific.

    PubMed

    Karayannis, Theofanis; De Marco García, Natalia V; Fishell, Gordon J

    2012-01-01

    Functional neuronal homeostasis has been studied in a variety of model systems and contexts. Many studies have shown that there are a number of changes that can be activated within individual cells or networks in order to compensate for perturbations or changes in levels of activity. Dissociating the cell autonomous from the network-mediated events has been complicated due to the difficulty of sparsely targeting specific populations of neurons in vivo. Here, we make use of a recent in vivo approach we developed that allows for the sparse labeling and manipulation of activity within superficial caudal ganglionic eminence (CGE)-derived GABAergic interneurons. Expression of the inward rectifying potassium channel Kir2.1 cell-autonomously reduced neuronal activity and lead to specific developmental changes in their intrinsic electrophysiological properties and the synaptic input they received. In contrast to previous studies on homeostatic scaling of pyramidal cells, we did not detect any of the typically observed compensatory mechanisms in these interneurons. Rather, we instead saw a specific alteration of the kinetics of excitatory synaptic events within the reelin-expressing subpopulation of interneurons. These results provide the first in vivo observations for the capacity of interneurons to cell-autonomously regulate their excitability. PMID:23015781

  19. A gender-specific measure of binge drinking among college students.

    PubMed Central

    Wechsler, H; Dowdall, G W; Davenport, A; Rimm, E B

    1995-01-01

    This study examined the relationship of volume of alcohol consumed to the occurrence of alcohol-related problems among male and female college students to develop a gender-specific measure of heavy episodic or binge drinking by college students for public health research. A self-administered survey was mailed to a national representative sample of students at 140 4-year colleges in 40 states and the District of Columbia. A total of 17,592 college students completed the survey. Women who typically drink four drinks in a row were found to have roughly the same likelihood of experiencing drinking-related problems as men who typically drink five drinks in a row. Use of the same standard for both sexes underestimates binge drinking and the negative health risks for women. PMID:7604925

  20. LPS Down-Regulates Specificity Protein 1 Activity by Activating NF-κB Pathway in Endotoxemic Mice

    PubMed Central

    Ye, Xiaobing; Liu, Hong; Gong, Yong-Sheng; Liu, Shu Fang

    2015-01-01

    Background Specificity protein (Sp) 1 mediates the transcription of a large number of constitutive genes encoding physiological mediators. NF-κB mediates the expression of hundreds of inducible genes encoding pathological mediators. Crosstalk between Sp1 and NF-κB pathways could be pathophysiologically significant, but has not been studied. This study examined the crosstalk between the two pathways and defined the role of NF-κB signaling in LPS-induced down-regulation of Sp1 activity. Methods and Main Findings Challenge of wild type mice with samonelia enteritidis LPS (10 mg/kg, i.p.) down-regulated Sp1 binding activity in lungs in a time-dependent manner, which was concomitantly associated with an increased NF-κB activity. LPS down-regulates Sp1 activity by inducing an LPS inducible Sp1-degrading enzyme (LISPDE) activity, which selectively degrades Sp1 protein, resulting in Sp1 down-regulation. Blockade of NF-κB activation in mice deficient in NF-κB p50 gene (NF-κB-KO) suppressed LISPDE activity, prevented Sp1 protein degradation, and reversed the down-regulation of Sp1 DNA binding activity and eNOS expression (an indicator of Sp1 transactivation activity). Inhibition of LISPDE activity using a selective LISPDE inhibitor mimicked the effects of NF-κB blockade. Pretreatment of LPS-challenged WT mice with a selective LISPDE inhibitor increased nuclear Sp1 protein content, restored Sp1 DNA binding activity and reversed eNOS protein down-regulation in lungs. Enhancing tissue level of Sp1 activity by inhibiting NF-κB-mediated Sp1 down-regulation increased tissue level of IL-10 and decreased tissue level of TNF- αin the lungs. Conclusions NF-κB signaling mediates LPS-induced down-regulation of Sp1 activity. Activation of NF-κB pathway suppresses Sp1 activity and Sp1-mediated anti-inflammatory signals. Conversely, Sp1 signaling counter-regulates NF-κB-mediated inflammatory response. Crosstalk between NF-κB and Sp1 pathways regulates the balance between pro

  1. Sensitivity of snow density and specific surface area measured by microtomography to different image processing algorithms

    NASA Astrophysics Data System (ADS)

    Hagenmuller, Pascal; Matzl, Margret; Chambon, Guillaume; Schneebeli, Martin

    2016-05-01

    Microtomography can measure the X-ray attenuation coefficient in a 3-D volume of snow with a spatial resolution of a few microns. In order to extract quantitative characteristics of the microstructure, such as the specific surface area (SSA), from these data, the greyscale image first needs to be segmented into a binary image of ice and air. Different numerical algorithms can then be used to compute the surface area of the binary image. In this paper, we report on the effect of commonly used segmentation and surface area computation techniques on the evaluation of density and specific surface area. The evaluation is based on a set of 38 X-ray tomographies of different snow samples without impregnation, scanned with an effective voxel size of 10 and 18 μm. We found that different surface area computation methods can induce relative variations up to 5 % in the density and SSA values. Regarding segmentation, similar results were obtained by sequential and energy-based approaches, provided the associated parameters were correctly chosen. The voxel size also appears to affect the values of density and SSA, but because images with the higher resolution also show the higher noise level, it was not possible to draw a definitive conclusion on this effect of resolution.

  2. Shape-specific activation of occipital cortex in an early blind echolocation expert.

    PubMed

    Arnott, Stephen R; Thaler, Lore; Milne, Jennifer L; Kish, Daniel; Goodale, Melvyn A

    2013-04-01

    We have previously reported that an early-blind echolocating individual (EB) showed robust occipital activation when he identified distant, silent objects based on echoes from his tongue clicks (Thaler, Arnott, & Goodale, 2011). In the present study we investigated the extent to which echolocation activation in EB's occipital cortex reflected general echolocation processing per se versus feature-specific processing. In the first experiment, echolocation audio sessions were captured with in-ear microphones in an anechoic chamber or hallway alcove as EB produced tongue clicks in front of a concave or flat object covered in aluminum foil or a cotton towel. All eight echolocation sessions (2 shapes×2 surface materials×2 environments) were then randomly presented to him during a sparse-temporal scanning fMRI session. While fMRI contrasts of chamber versus alcove-recorded echolocation stimuli underscored the importance of auditory cortex for extracting echo information, main task comparisons demonstrated a prominent role of occipital cortex in shape-specific echo processing in a manner consistent with latent, multisensory cortical specialization. Specifically, relative to surface composition judgments, shape judgments elicited greater BOLD activity in ventrolateral occipital areas and bilateral occipital pole. A second echolocation experiment involving shape judgments of objects located 20° to the left or right of straight ahead activated more rostral areas of EB's calcarine cortex relative to location judgments of those same objects and, as we previously reported, such calcarine activity was largest when the object was located in contralateral hemispace. Interestingly, other echolocating experts (i.e., a congenitally blind individual in Experiment 1, and a late blind individual in Experiment 2) did not show the same pattern of feature-specific echo-processing calcarine activity as EB, suggesting the possible significance of early visual experience and early

  3. Biophysical and mutagenic analysis of Thermoanaerobacter ethanolicus secondary-alcohol dehydrogenase activity and specificity.

    PubMed Central

    Burdette, D S; Secundo, F; Phillips, R S; Dong, J; Scott, R A; Zeikus, J G

    1997-01-01

    The Thermoanaerobacter ethanolicus 39E adhB gene encoding the secondary-alcohol dehydrogenase (secondary ADH) was overexpressed in Escherichia coli at more than 10% of total protein. The recombinant enzyme was purified in high yield (67%) by heat-treatment at 85 degrees C and (NH4)2SO4 precipitation. Site-directed mutants (C37S, H59N, D150N, D150Eand D150C were analysed to test the peptide sequence comparison-based predictions of amino acids responsible for putative catalytic Zn binding. X-ray absorption spectroscopy confirmed the presence of a protein-bound Zn atom with ZnS1(imid)1(N,O)3 co-ordination sphere. Inductively coupled plasma atomic emission spectrometry measured 0.48 Zn atoms per wild-type secondary ADH subunit. The C37S, H59N and D150N mutant enzymes bound only 0.11, 0.13 and 0.33 Zn per subunit respectively,suggesting that these residues are involved in Zn liganding. The D150E and D150C mutants retained 0.47 and 1.2 Zn atoms per subunit, indicating that an anionic side-chain moiety at this position preserves the bound Zn. All five mutant enzymes had activity, suggesting that the T. ethanolicus secondary ADH requires a properly co-ordinated catalytic Zn atom. The His-59 and Asp-150 mutations also altered secondary ADH affinity for propan-2-ol over a 140-fold range, whereas the overall change in affinity for ethanol spanned a range of only 7-fold, supporting the importance of the metal in secondary ADH substrate binding. The lack of significant changes in cofactor affinity as a result of these catalytic Zn ligand mutations suggested that secondary ADH substrate-and cofactor-binding sites are structurally distinct. Altering Gly198 to Asp reduced the enzyme specific activity 2.7-fold, increased the Km(app) for NADP+ 225-fold, and decreased the Km(app) for NAD+ 3-fold, supporting the prediction that the enzyme binds nicotinamide cofactor in a Rossmann fold. Our data indicate therefore that, unlike the liver primary ADH

  4. Behavioral state-specific inhibitory postsynaptic potentials impinge on cat lumbar motoneurons during active sleep.

    PubMed

    Morales, F R; Boxer, P; Chase, M H

    1987-11-01

    High-gain intracellular records were obtained from lumbar motoneurons in intact, undrugged cats during naturally occurring states of wakefulness, quiet sleep, and active sleep. Spontaneous, discrete, inhibitory postsynaptic potentials (IPSPs) were found to impinge on lumbar motoneurons during all states of sleep and wakefulness. IPSPs which occurred during wakefulness and quiet sleep were of relatively low amplitude and had a low frequency of occurrence. During the state of active sleep there occurred a great increase in inhibitory input. This was the result of the appearance of large-amplitude IPSPs and of an increase in the frequency of low-amplitude IPSPs which were indistinguishable from those recorded during wakefulness and quiet sleep. In addition to a difference in amplitude, the time course of the large IPSPs recorded during active sleep further differentiated them from the smaller IPSPs recorded during wakefulness, quiet sleep, and active sleep; i.e., their rise-time and half-width were of longer duration and their rate-of-rise was significantly faster. We suggest that the large, active sleep-specific IPSPs reflect the activity of a group of inhibitory interneurons which are inactive during wakefulness and quiet sleep and which discharge during active sleep. These as yet unidentified interneurons would then serve as the last link in the brain stem-spinal cord inhibitory system which is responsible for producing muscle atonia during the state of active sleep. PMID:3666087

  5. Neural activity promotes long-distance, target-specific regeneration of adult retinal axons.

    PubMed

    Lim, Jung-Hwan A; Stafford, Benjamin K; Nguyen, Phong L; Lien, Brian V; Wang, Chen; Zukor, Katherine; He, Zhigang; Huberman, Andrew D

    2016-08-01

    Axons in the mammalian CNS fail to regenerate after injury. Here we show that if the activity of mouse retinal ganglion cells (RGCs) is increased by visual stimulation or using chemogenetics, their axons regenerate. We also show that if enhancement of neural activity is combined with elevation of the cell-growth-promoting pathway involving mammalian target of rapamycin (mTOR), RGC axons regenerate long distances and re-innervate the brain. Analysis of genetically labeled RGCs revealed that this regrowth can be target specific: RGC axons navigated back to their correct visual targets and avoided targets incorrect for their function. Moreover, these regenerated connections were successful in partially rescuing a subset of visual behaviors. Our findings indicate that combining neural activity with activation of mTOR can serve as powerful tool for enhancing axon regeneration, and they highlight the remarkable capacity of CNS neurons to re-establish accurate circuit connections in adulthood. PMID:27399843

  6. Tissue-specific activity of two manganese superoxide dismutase promoters in transgenic tobacco.

    PubMed Central

    Van Camp, W; Hérouart, D; Willekens, H; Takahashi, H; Saito, K; Van Montagu, M; Inzé, D

    1996-01-01

    In eukaryotes, manganese superoxide dismutase is a nuclear-encoded protein that scavenges superoxide radicals in the mitochondrial matrix. We have isolated two manganese superoxide dismutase genes from Nicotiana plumbaginifolia L. and fused the 5' upstream regulatory region of these genes to the beta-glucuronidase reporter gene. The two gene fusions displayed a differential tissue specificity in transgenic tobacco (Nicotiana tabacum). Promoter activity of the SodA1 gene fusion was found in the pollen, middle layer, and stomium of anthers, but was usually undetectable in vegetative organs of mature plants. The SodA2 gene fusion was expressed in the leaves, stems, roots, and flowers. SodA2 promoter activity was most prominent in the vascular bundles, stomata, axillary buds, pericycle, stomium, and pollen. Histochemical analysis of succinate dehydrogenase activity suggested that the spatial expression of the two gene fusions is generally correlated with mitochondrial respiratory activity. PMID:8883376

  7. Can Specific Protein-Lipid Interactions Stabilize an Active State of the Beta 2 Adrenergic Receptor?

    PubMed

    Neale, Chris; Herce, Henry D; Pomès, Régis; García, Angel E

    2015-10-20

    G-protein-coupled receptors are eukaryotic membrane proteins with broad biological and pharmacological relevance. Like all membrane-embedded proteins, their location and orientation are influenced by lipids, which can also impact protein function via specific interactions. Extensive simulations totaling 0.25 ms reveal a process in which phospholipids from the membrane's cytosolic leaflet enter the empty G-protein binding site of an activated β2 adrenergic receptor and form salt-bridge interactions that inhibit ionic lock formation and prolong active-state residency. Simulations of the receptor embedded in an anionic membrane show increased lipid binding, providing a molecular mechanism for the experimental observation that anionic lipids can enhance receptor activity. Conservation of the arginine component of the ionic lock among Rhodopsin-like G-protein-coupled receptors suggests that intracellular lipid ingression between receptor helices H6 and H7 may be a general mechanism for active-state stabilization. PMID:26488656

  8. Print-specific multimodal brain activation in kindergarten improves prediction of reading skills in second grade.

    PubMed

    Bach, Silvia; Richardson, Ulla; Brandeis, Daniel; Martin, Ernst; Brem, Silvia

    2013-11-15

    Children who are poor readers usually experience troublesome school careers and consequently often suffer from secondary emotional and behavioural problems. Early identification and prediction of later reading problems thus are critical in order to start targeted interventions for those children with an elevated risk for emerging reading problems. In this study, behavioural precursors of reading were assessed in nineteen (aged 6.4 ± 0.3 years) non-reading kindergarteners before training letter-speech sound associations with a computerized game (Graphogame) for eight weeks. The training aimed to introduce the basic principles of letter-speech sound correspondences and to initialize the sensitization of specific brain areas to print. Event-related potentials (ERP) and functional magnetic resonance imaging (fMRI) data were recorded during an explicit word/symbol processing task after the training. Reading skills were assessed two years later in second grade. The focus of this study was on clarifying whether electrophysiological and fMRI data of kindergarten children significantly improve prediction of future reading skills in 2nd grade over behavioural data alone. Based on evidence from previous studies demonstrating the importance of initial print sensitivity in the left occipito-temporal visual word form system (VWFS) for learning to read, the first pronounced difference in processing words compared to symbols in the ERP, an occipito-temporal negativity (N1: 188-281 ms) along with the corresponding functional activation in the left occipito-temporal VWFS were defined as potential predictors. ERP and fMRI data in kindergarteners significantly improved the prediction of reading skills in 2nd grade over behavioural data alone. Together with the behavioural measures they explained up to 88% of the variance. An additional discriminant analysis revealed a remarkably high accuracy in classifying normal (n=11) and poor readers (n=6). Due to the key limitation of the study

  9. Evaluation of 16 measures of mental workload using a simulated flight task emphasizing mediational activity

    NASA Technical Reports Server (NTRS)

    Wierwille, W. W.; Rahimi, M.; Casali, J. G.

    1985-01-01

    As aircraft and other systems become more automated, a shift is occurring in human operator participation in these systems. This shift is away from manual control and toward activities that tap the higher mental functioning of human operators. Therefore, an experiment was performed in a moving-base flight simulator to assess mediational (cognitive) workload measurement. Specifically, 16 workload estimation techniques were evaluated as to their sensitivity and intrusion in a flight task emphasizing mediational behavior. Task loading, using navigation problems presented on a display, was treated as an independent variable, and workload-measure values were treated as dependent variables. Results indicate that two mediational task measures, two rating scale measures, time estimation, and two eye behavior measures were reliably sensitive to mediational loading. The time estimation measure did, however, intrude on mediational task performance. Several of the remaining measures were completely insensitive to mediational load.

  10. The Chinese Hamster Dihydrofolate Reductase Replication Origin Beta Is Active at Multiple Ectopic Chromosomal Locations and Requires Specific DNA Sequence Elements for Activity

    PubMed Central

    Altman, Amy L.; Fanning, Ellen

    2001-01-01

    To identify cis-acting genetic elements essential for mammalian chromosomal DNA replication, a 5.8-kb fragment from the Chinese hamster dihydrofolate reductase (DHFR) locus containing the origin beta (ori-β) initiation region was stably transfected into random ectopic chromosomal locations in a hamster cell line lacking the endogenous DHFR locus. Initiation at ectopic ori-β in uncloned pools of transfected cells was measured using a competitive PCR-based nascent strand abundance assay and shown to mimic that at the endogenous ori-β region in Chinese hamster ovary K1 cells. Initiation activity of three ectopic ori-β deletion mutants was reduced, while the activity of another deletion mutant was enhanced. The results suggest that a 5.8-kb fragment of the DHFR ori-β region is sufficient to direct initiation and that specific DNA sequences in the ori-β region are required for efficient initiation activity. PMID:11158297

  11. Exploring Patient Activation in the Clinic: Measurement from Three Perspectives

    ERIC Educational Resources Information Center

    Ledford, Christy J. W.; Ledford, Christopher C.; Childress, Marc A.

    2013-01-01

    Objective. To further conceptualize and operationalize patient activation (PA), using measures from patient, physician, and researcher perspectives. Data Source/Study Setting. Multimethod observation in 2010 within a family medicine clinic. Study Design. Part of an intervention with 130 patients with type 2 diabetes, this observational study…

  12. Measuring Perceived Benefits and Perceived Barriers for Physical Activity

    ERIC Educational Resources Information Center

    Brown, Seth A.

    2005-01-01

    Objectives: To evaluate the psychometric properties and relationship to physical activity levels of the Exercise Benefits/Barriers Scale (EBBS) among college students. Methods: A total of 398 college students completed the EBBS and a measure of self-efficacy, the Physical Exercise Self-Efficacy Scale. In addition, a subsample of 275 students also…

  13. Measurement of the Construct of Attitude Toward Student Activism.

    ERIC Educational Resources Information Center

    Gold, Joel A.; And Others

    The study measures individual differences in attitude toward student activism on an 18-item Likert type scale. A validity comparison of a known extreme group with a sample from the general student population yielded a significant difference between the means of the groups. Construct validity was evaluated by first testing the differences between…

  14. A Self-Report Measure of Physical Activity

    ERIC Educational Resources Information Center

    Siegel, Donald

    2005-01-01

    There are multiple approaches to measuring physical activity. Among these are direct observation, electronic monitoring, direct and indirect calorimetry, and self-report instruments. Self-report instruments are the most practical and cost effective option for use with a large group. In a study by Motl, Dishman, Dowda, and Pate (2004), two groups…

  15. Measurement for Work. Teaching Guide and Sample Learning Activities.

    ERIC Educational Resources Information Center

    Angel, Margo; Bolton, Chris

    This document is intended to help Australian technical and further education instructors in New South Wales (TAFE NSW) identify teaching principles and learning activities that they can use to help adult learners master the mathematics processes, knowledge, and skills needed to perform basic measurement tasks in today's workplace. The materials…

  16. Measurement of Perceived School Climate for Active Travel in Children

    ERIC Educational Resources Information Center

    Evenson, Kelly R.; Motl, Robert W.; Birnbaum, Amanda S.; Ward, Dianne S.

    2007-01-01

    Objectives: To describe the development of an original scale that measures perceived school climate for active travel in fourth- and fifth-grade girls and boys. Methods: The data were analyzed using confirmatory factor analysis (CFA) to provide evidence of factorial validity, factorial invariance, and construct validity. Results: The CFA supported…

  17. Specific induction of endogenous viral restriction factors using CRISPR/Cas-derived transcriptional activators

    PubMed Central

    Bogerd, Hal P.; Kornepati, Anand V. R.; Marshall, Joy B.; Kennedy, Edward M.; Cullen, Bryan R.

    2015-01-01

    Whereas several mammalian proteins can restrict the replication of HIV-1 and other viruses, these are often not expressed in relevant target cells. A potential method to inhibit viral replication might therefore be to use synthetic transcription factors to induce restriction factor expression. In particular, mutants of the RNA-guided DNA binding protein Cas9 that have lost their DNA cleavage activity could be used to recruit transcription activation domains to specific promoters. However, initial experiments revealed only weak activation unless multiple promoter-specific single guide RNAs (sgRNAs) were used. Recently, the recruitment of multiple transcription activation domains by a single sgRNA, modified to contain MS2-derived stem loops that recruit fusion proteins consisting of the MS2 coat protein linked to transcription activation domains, was reported to induce otherwise silent cellular genes. Here, we demonstrate that such “synergistic activation mediators” can induce the expression of two restriction factors, APOBEC3G (A3G) and APOBEC3B (A3B), in human cells that normally lack these proteins. We observed modest activation of endogenous A3G or A3B expression using single sgRNAs but high expression when two sgRNAs were used. Whereas the induced A3G and A3B proteins both blocked infection by an HIV-1 variant lacking a functional vif gene by inducing extensive dC-to-dU editing, only the induced A3B protein inhibited wild-type HIV-1. These data demonstrate that Cas9-derived transcriptional activators have the potential to be used for screens for endogenous genes that affect virus replication and raise the possibility that synthetic transcription factors might prove clinically useful if efficient delivery mechanisms could be developed. PMID:26668372

  18. Specific induction of endogenous viral restriction factors using CRISPR/Cas-derived transcriptional activators.

    PubMed

    Bogerd, Hal P; Kornepati, Anand V R; Marshall, Joy B; Kennedy, Edward M; Cullen, Bryan R

    2015-12-29

    Whereas several mammalian proteins can restrict the replication of HIV-1 and other viruses, these are often not expressed in relevant target cells. A potential method to inhibit viral replication might therefore be to use synthetic transcription factors to induce restriction factor expression. In particular, mutants of the RNA-guided DNA binding protein Cas9 that have lost their DNA cleavage activity could be used to recruit transcription activation domains to specific promoters. However, initial experiments revealed only weak activation unless multiple promoter-specific single guide RNAs (sgRNAs) were used. Recently, the recruitment of multiple transcription activation domains by a single sgRNA, modified to contain MS2-derived stem loops that recruit fusion proteins consisting of the MS2 coat protein linked to transcription activation domains, was reported to induce otherwise silent cellular genes. Here, we demonstrate that such "synergistic activation mediators" can induce the expression of two restriction factors, APOBEC3G (A3G) and APOBEC3B (A3B), in human cells that normally lack these proteins. We observed modest activation of endogenous A3G or A3B expression using single sgRNAs but high expression when two sgRNAs were used. Whereas the induced A3G and A3B proteins both blocked infection by an HIV-1 variant lacking a functional vif gene by inducing extensive dC-to-dU editing, only the induced A3B protein inhibited wild-type HIV-1. These data demonstrate that Cas9-derived transcriptional activators have the potential to be used for screens for endogenous genes that affect virus replication and raise the possibility that synthetic transcription factors might prove clinically useful if efficient delivery mechanisms could be developed. PMID:26668372

  19. Diversity between mammalian tolloid proteinases: Oligomerisation and non-catalytic domains influence activity and specificity

    PubMed Central

    Bayley, Christopher P.; Ruiz Nivia, Hilda D.; Dajani, Rana; Jowitt, Thomas A.; Collins, Richard F.; Rada, Heather; Bird, Louise E.; Baldock, Clair

    2016-01-01

    The mammalian tolloid family of metalloproteinases is essential for tissue patterning and extracellular matrix assembly. The four members of the family: bone morphogenetic protein-1 (BMP-1), mammalian tolloid (mTLD), tolloid-like (TLL)-1 and TLL-2 differ in their substrate specificity and activity levels, despite sharing similar domain organization. We have previously described a model of substrate exclusion by dimerisation to explain differences in the activities of monomeric BMP-1 and dimers of mTLD and TLL-1. Here we show that TLL-2, the least active member of the tolloid family, is predominantly monomeric in solution, therefore it appears unlikely that substrate exclusion via dimerisation is a mechanism for regulating TLL-2 activity. X-ray scattering and electron microscopy structural and biophysical analyses reveal an elongated shape for the monomer and flexibility in the absence of calcium. Furthermore, we show that TLL-2 can cleave chordin in vitro, similar to other mammalian tolloids, but truncated forms of TLL-2 mimicking BMP-1 are unable to cleave chordin. However, both the N- and C-terminal non-catalytic domains from all mammalian tolloids bind chordin with high affinity. The mechanisms underlying substrate specificity and activity in the tolloid family are complex with variation between family members and depend on both multimerisation and substrate interaction. PMID:26902455

  20. A flower-specific Myb protein activates transcription of phenylpropanoid biosynthetic genes.

    PubMed

    Sablowski, R W; Moyano, E; Culianez-Macia, F A; Schuch, W; Martin, C; Bevan, M

    1994-01-01

    Synthesis of flavonoid pigments in flowers requires the co-ordinated expression of genes encoding enzymes in th phenylpropanoid biosynthetic pathway. Some cis-elements involved in the transcriptional control of these genes have been defined. We report binding of petal-specific activities from tobacco and Antirrhinum majus (snapdragon) to an element conserved in promoters of phenylpropanoid biosynthetic genes and implicated in expression in flowers. These binding activities were inhibited by antibodies raised against Myb305, a flower-specific Myb protein previously cloned from Antirrhinum by sequence homology. Myb305 bound to the same element and formed a DNA-protein complex with the same mobility as the Antirrhinum petal protein in electrophoretic mobility shift experiments. Myb305 activated expression from its binding site in yeast and in tobacco protoplasts. In protoplasts, activation also required a G-box-like element, suggesting co-operation with other elements and factors. The results strongly suggest a role for Myb305-related proteins in the activation of phenylpropanoid biosynthetic genes in flowers. This is consistent with the genetically demonstrated role of plant Myb proteins in the regulation of genes involved in flavonoid synthesis. PMID:8306956

  1. Cardiac myocyte-specific AHR activation phenocopies TCDD-induced toxicity in zebrafish.

    PubMed

    Lanham, Kevin A; Plavicki, Jessica; Peterson, Richard E; Heideman, Warren

    2014-09-01

    Exposure of zebrafish embryos to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) activates the zebrafish aryl hydrocarbon receptor 2 (AHR) to produce developmental and cardiovascular toxicity. AHR is found in the heart; however, AHR activation by TCDD is not confined to the heart and occurs throughout the organism. In order to understand the cause of cardiotoxicity, we constructed a constitutively active AHR (caAHR) based on the zebrafish AHR2 and expressed it specifically in cardiomyocytes. We show that AHR activation within the cardiomyocytes can account for the heart failure induced by TCDD. Expression of the caAHR within the heart produced cardiac malformations, loss of circulation, and pericardial edema. The heart-specific activation of AHR reproduced several other well-characterized endpoints of TCDD toxicity outside of the cardiovascular system, including defects in swim bladder and craniofacial development. This work identifies a single cellular site of TCDD action, the myocardial cell, that can account for the severe cardiovascular collapse observed following early life stage exposure to TCDD, and contributes to other forms of toxicity. PMID:25037585

  2. Category-specific activations during word generation reflect experiential sensorimotor modalities

    PubMed Central

    Hwang, Kai; Palmer, Erica D.; Basho, Surina; Zadra, Jonathan R.; Müller, Ralph-Axel

    2009-01-01

    According to the sensorimotor theory of lexicosemantic organization, semantic representations are neurally distributed and anatomically linked to category-specific sensory areas. Previous functional neuroimaging studies have demonstrated category specificity in lexicosemantic representations. However, little evidence is available from word generation paradigms, which provide access to semantic representations while minimizing confounds resulting from low-level perceptual features of stimulus presentation. In this study, 13 healthy young adults underwent fMRI scanning while performing a word generation task, generating exemplars to nine different semantic categories. Each semantic category was assigned to one of three superordinate category types, based upon sensorimotor modalities (visual, motor, somatosensory) presumed to predominate in lexical acquisition. For word generation overall, robust activation was seen in left inferior frontal cortex. Analyses by sensorimotor modality categories yielded activations in brain regions related to perceptual and motor processing: Visual categories activated extrastriate cortex, motor categories activated the intraparietal sulcus and posterior middle temporal cortex, and somatosensory categories activated postcentral and inferior parietal regions. Our results are consistent with the sensorimotor theory, according to which lexicosemantic representations are distributed across brain regions participating in sensorimotor processing associated with the experiential components of lexicosemantic acquisition. PMID:19559802

  3. Site-specific Interaction Mapping of Phosphorylated Ubiquitin to Uncover Parkin Activation.

    PubMed

    Yamano, Koji; Queliconi, Bruno B; Koyano, Fumika; Saeki, Yasushi; Hirokawa, Takatsugu; Tanaka, Keiji; Matsuda, Noriyuki

    2015-10-16

    Damaged mitochondria are eliminated through autophagy machinery. A cytosolic E3 ubiquitin ligase Parkin, a gene product mutated in familial Parkinsonism, is essential for this pathway. Recent progress has revealed that phosphorylation of both Parkin and ubiquitin at Ser(65) by PINK1 are crucial for activation and recruitment of Parkin to the damaged mitochondria. However, the mechanism by which phosphorylated ubiquitin associates with and activates phosphorylated Parkin E3 ligase activity remains largely unknown. Here, we analyze interactions between phosphorylated forms of both Parkin and ubiquitin at a spatial resolution of the amino acid residue by site-specific photo-crosslinking. We reveal that the in-between-RING (IBR) domain along with RING1 domain of Parkin preferentially binds to ubiquitin in a phosphorylation-dependent manner. Furthermore, another approach, the Fluoppi (fluorescent-based technology detecting protein-protein interaction) assay, also showed that pathogenic mutations in these domains blocked interactions with phosphomimetic ubiquitin in mammalian cells. Molecular modeling based on the site-specific photo-crosslinking interaction map combined with mass spectrometry strongly suggests that a novel binding mechanism between Parkin and ubiquitin leads to a Parkin conformational change with subsequent activation of Parkin E3 ligase activity. PMID:26260794

  4. Cardiac Myocyte-Specific AHR Activation Phenocopies TCDD-Induced Toxicity in Zebrafish

    PubMed Central

    Lanham, Kevin A.; Plavicki, Jessica; Peterson, Richard E.; Heideman, Warren

    2014-01-01

    Exposure of zebrafish embryos to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) activates the zebrafish aryl hydrocarbon receptor 2 (AHR) to produce developmental and cardiovascular toxicity. AHR is found in the heart; however, AHR activation by TCDD is not confined to the heart and occurs throughout the organism. In order to understand the cause of cardiotoxicity, we constructed a constitutively active AHR (caAHR) based on the zebrafish AHR2 and expressed it specifically in cardiomyocytes. We show that AHR activation within the cardiomyocytes can account for the heart failure induced by TCDD. Expression of the caAHR within the heart produced cardiac malformations, loss of circulation, and pericardial edema. The heart-specific activation of AHR reproduced several other well-characterized endpoints of TCDD toxicity outside of the cardiovascular system, including defects in swim bladder and craniofacial development. This work identifies a single cellular site of TCDD action, the myocardial cell, that can account for the severe cardiovascular collapse observed following early life stage exposure to TCDD, and contributes to other forms of toxicity. PMID:25037585

  5. An activity-based probe for high-throughput measurements of triacylglycerol lipases.

    PubMed

    Tam, John; Henault, Martin; Li, Lianhai; Wang, Zhaoyin; Partridge, Anthony W; Melnyk, Roman A

    2011-07-15

    Modulating the activity of lipases involved in the metabolism of plasma lipoproteins is an attractive approach for developing lipid raising/lowering therapies to treat cardiovascular disease. Identifying small molecule inhibitors for these membrane-active enzymes, however, is complicated by difficulties associated with measuring lipase activity and inhibition at the water-membrane interface; substrate and compound dynamics at the particle interface have the potential to confound data interpretation. Here, we describe a novel ELISA-based lipase activity assay that employs as "bait" a biotinylated active-site probe that irreversibly binds to the catalytic active-site serine of members of the triacylglycerol lipase family (hepatic lipase, lipoprotein lipase, and endothelial lipase) in solution with high affinity. Detection of "captured" (probe-enzyme) complexes on streptavidin-coated plates using labeled secondary antibodies to specific primary antibodies offers several advantages over conventional assays, including the ability to eliminate enzyme-particle and compound-particle effects; specifically measure lipase activity in complex mixtures in vitro; preferentially identify active-site-directed inhibitors; and distinguish between reversible and irreversible inhibitors through a simple assay modification. Using EL as an exemplar, we demonstrate the versatility of this assay both for high-throughput screening and for compound mechanism-of-action studies. PMID:21397586

  6. Synthesis of Isomeric Phosphoubiquitin Chains Reveals that Phosphorylation Controls Deubiquitinase Activity and Specificity.

    PubMed

    Huguenin-Dezot, Nicolas; De Cesare, Virginia; Peltier, Julien; Knebel, Axel; Kristaryianto, Yosua Adi; Rogerson, Daniel T; Kulathu, Yogesh; Trost, Matthias; Chin, Jason W

    2016-07-26

    Ubiquitin is post-translationally modified by phosphorylation at several sites, but the consequences of these modifications are largely unknown. Here, we synthesize multi-milligram quantities of ubiquitin phosphorylated at serine 20, serine 57, and serine 65 via genetic code expansion. We use these phosphoubiquitins for the enzymatic assembly of 20 isomeric phosphoubiquitin dimers, with different sites of isopeptide linkage and/or phosphorylation. We discover that phosphorylation of serine 20 on ubiquitin converts UBE3C from a dual-specificity E3 ligase into a ligase that primarily synthesizes K48 chains. We profile the activity of 31 deubiquitinases on the isomeric phosphoubiquitin dimers in 837 reactions, and we discover that phosphorylation at distinct sites in ubiquitin can activate or repress cleavage of a particular linkage by deubiquitinases and that phosphorylation at a single site in ubiquitin can control the specificity of deubiquitinases for distinct ubiquitin linkages. PMID:27425610

  7. Physiological characterization of the ARO10-dependent, broad-substrate-specificity 2-oxo acid decarboxylase activity of Saccharomyces cerevisiae.

    PubMed

    Vuralhan, Zeynep; Luttik, Marijke A H; Tai, Siew Leng; Boer, Viktor M; Morais, Marcos A; Schipper, Dick; Almering, Marinka J H; Kötter, Peter; Dickinson, J Richard; Daran, Jean-Marc; Pronk, Jack T

    2005-06-01

    Aerobic, glucose-limited chemostat cultures of Saccharomyces cerevisiae CEN.PK113-7D were grown with different nitrogen sources. Cultures grown with phenylalanine, leucine, or methionine as a nitrogen source contained high levels of the corresponding fusel alcohols and organic acids, indicating activity of the Ehrlich pathway. Also, fusel alcohols derived from the other two amino acids were detected in the supernatant, suggesting the involvement of a common enzyme activity. Transcript level analysis revealed that among the five thiamine-pyrophospate-dependent decarboxylases (PDC1, PDC5, PDC6, ARO10, and THI3), only ARO10 was transcriptionally up-regulated when phenylalanine, leucine, or methionine was used as a nitrogen source compared to growth on ammonia, proline, and asparagine. Moreover, 2-oxo acid decarboxylase activity measured in cell extract from CEN.PK113-7D grown with phenylalanine, methionine, or leucine displayed similar broad-substrate 2-oxo acid decarboxylase activity. Constitutive expression of ARO10 in ethanol-limited chemostat cultures in a strain lacking the five thiamine-pyrophosphate-dependent decarboxylases, grown with ammonia as a nitrogen source, led to a measurable decarboxylase activity with phenylalanine-, leucine-, and methionine-derived 2-oxo acids. Moreover, even with ammonia as the nitrogen source, these cultures produced significant amounts of the corresponding fusel alcohols. Nonetheless, the constitutive expression of ARO10 in an isogenic wild-type strain grown in a glucose-limited chemostat with ammonia did not lead to any 2-oxo acid decarboxylase activity. Furthermore, even when ARO10 was constitutively expressed, growth with phenylalanine as the nitrogen source led to increased decarboxylase activities in cell extracts. The results reported here indicate the involvement of posttranscriptional regulation and/or a second protein in the ARO10-dependent, broad-substrate-specificity decarboxylase activity. PMID:15933030

  8. Vehicle-specific emissions modeling based upon on-road measurements.

    PubMed

    Frey, H Christopher; Zhang, Kaishan; Rouphail, Nagui M

    2010-05-01

    Vehicle-specific microscale fuel use and emissions rate models are developed based upon real-world hot-stabilized tailpipe measurements made using a portable emissions measurement system. Consecutive averaging periods of one to three multiples of the response time are used to compare two semiempirical physically based modeling schemes. One scheme is based on internally observable variables (IOVs), such as engine speed and manifold absolute pressure, while the other is based on externally observable variables (EOVs), such as speed, acceleration, and road grade. For NO, HC, and CO emission rates, the average R(2) ranged from 0.41 to 0.66 for the former and from 0.17 to 0.30 for the latter. The EOV models have R(2) for CO(2) of 0.43 to 0.79 versus 0.99 for the IOV models. The models are sensitive to episodic events in driving cycles such as high acceleration. Intervehicle and fleet average modeling approaches are compared; the former account for microscale variations that might be useful for some types of assessments. EOV-based models have practical value for traffic management or simulation applications since IOVs usually are not available or not used for emission estimation. PMID:20377172

  9. Lysine-specific demethylase-1 contributes to malignant behavior by regulation of invasive activity and metabolic shift in esophageal cancer.

    PubMed

    Kosumi, Keisuke; Baba, Yoshifumi; Sakamoto, Akihisa; Ishimoto, Takatsugu; Harada, Kazuto; Nakamura, Kenichi; Kurashige, Junji; Hiyoshi, Yukiharu; Iwatsuki, Masaaki; Iwagami, Shiro; Sakamoto, Yasuo; Miyamoto, Yuji; Yoshida, Naoya; Oki, Eiji; Watanabe, Masayuki; Hino, Shinjiro; Nakao, Mitsuyoshi; Baba, Hideo

    2016-01-15

    Lysine-specific demethylase-1 (LSD1) removes the methyl groups from mono- and di-methylated lysine 4 of histone H3. Previous studies have linked LSD1 to malignancy in several human tumors, and LSD1 is considered to epigenetically regulate the energy metabolism genes in adipocytes and hepatocellular carcinoma. This study investigates the function of LSD1 in the invasive activity and the metabolism of esophageal cancer cells. We investigated whether LSD1 immunohistochemical expression levels are related to clinical and pathological features, including the maximum standard uptake value in fluorodeoxyglucose positron emission tomography assay. The influence of LSD1 on cell proliferation, invasion and glucose uptake was evaluated in vitro by using specific small interfering RNA for LSD1, and an LSD1 inhibitor. We also evaluated two major energy pathways (glycolytic pathway and mitochondrial respiration) by measuring the extracellular acidification rate (ECAR) and the oxygen consumption rate (OCR) with an extracellular flux analyzer. High LSD1 immunohistochemical expression was significantly associated with high tumor stage, lymphovascular invasion, poor prognosis, and high maximum standard uptake value in esophageal cancer patients. In the in vitro analysis, LSD1 knockdown significantly suppressed the invasive activity and glucose uptake of cancerous cells, reduced their ECAR and increased their OCR and OCR/ECAR. LSD1 may contribute to malignant behavior by regulating the invasive activity and metabolism, activating the glycolytic pathway and inhibiting the mitochondrial respiration of esophageal cancer cells. The results support LSD1 as a potential therapeutic target. PMID:26240060

  10. Positional specificity of a phospholipase A activity induced by wounding, systemin, and oligosaccharide elicitors in tomato leaves

    PubMed Central

    Narvaez-Vasquez, J; Florin-Christensen, J; Ryan, CA

    1999-01-01

    Phospholipase A (PLA) activity, as measured by the accumulation of (14)C-lysophosphatidylcholine in leaves of tomato plants, increased rapidly and systemically in response to wounding. The increase in PLA activity in the systemic unwounded leaves was biphasic in wild-type tomato plants, peaking at 15 min and again at 60 min, but the second peak of activity was absent in transgenic prosystemin antisense plants. Supplying young excised tomato plants with the polypeptide hormone systemin also caused (14)C-lysophosphatidylcholine to increase to levels similar to those induced by wounding, but the increase in activity persisted for >2 hr. Antagonists of systemin blocked both the release of (14)C-lysophosphatidylcholine and the accumulation of defense proteins in response to systemin. (14)C-lysophosphatidylcholine levels did not increase in response to jasmonic acid. Chemical acylation of the lysophosphatidylcholine produced by wounding, systemin, and oligosaccharide elicitors followed by enzymatic hydrolysis with lipases of known specificities demostrated that the lysophosphatidylcholine is generated by a PLA with specificity for the sn-2 position. PMID:10559447

  11. Specific NFκB subunit activation and kinetics of cytokine induction in adenoviral keratitis

    PubMed Central

    Rajaiya, Jaya; Sadeghi, Neda

    2009-01-01

    Purpose Corneal inflammation associated with ocular adenoviral infection is caused by leukocytic infiltration of the subepithelial stroma in response to expression of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) by infected corneal cells. We have shown that these two chemokines are activated by the mitogen-activated protein kinases (MAPKs) extracellular signal-regulated kinase (ERK) and p38 for IL-8, and Jun-terminal kinase (JNK) for MCP-1. It is also well established that transcription of each of these chemokines is tightly controlled by the nuclear factor kappa B (NFκB) transcription factor family. Therefore, we sought to better understand the differential regulation of chemokine expression by NFκB in adenoviral infection of the cornea. Methods Primary keratocytes derived from human donor corneas were treated with signaling inhibitors and small interfering RNA specific to MAPKs, and infected with adenovirus for different time periods before analysis. Activation of specific NFκB subunits was analyzed by western blot, confocal microscopy, electromobility shift assay, and chromatin immunoprecipitation, and chemokine expression was quantified by enzyme-linked immunosorbent assay. Results Upon adenoviral infection, NFκB p65, p50, and cREL subunits translocate to the nucleus. This translocation is blocked by inhibitors of specific MAPK signaling pathways. Confocal microscopy showed that inhibitors of the p38, JNK, and ERK pathways differentially inhibited NFκB nuclear translocation, while PP2, an inhibitor of Src family kinases, completely inhibited NFκB nuclear translocation. Western blot analysis revealed that activation of specific NFκB subunits was time dependent following infection. Chromatin immunoprecipitation experiments indicated that binding of NFκB p65 and p50 subunits to the IL-8 promoter upon viral infection was differentially reduced by chemical inhibitors of MAPKs. Electromobility shift assay and luciferase assay analysis

  12. Methods for Specific Electrode Resistance Measurement during Transcranial Direct Current Stimulation

    PubMed Central

    Khadka, Niranjan; Rahman, Asif; Sarantos, Chris; Truong, Dennis Q.; Bikson, Marom

    2014-01-01

    Background Transcranial Direct Current Stimulation (tDCS) is investigated to treat a wide range of neuropsychiatric disorders, for rehabilitation, and for enhancing cognitive performance. The monitoring of electrode resistance before and during tDCS is considered important for tolerability and safety, where an unusually high resistance is indicative of undesired electrode or poor skin contact conditions. Conventional resistance measurement methods do not isolate individual electrode resistance but rather measures overall voltage. Moreover, for HD-tDCS devices, cross talk across electrodes makes concurrent resistance monitoring unreliable. Objective We propose a novel method for monitoring of the individual electrode resistance during tDCS, using a super-position of direct current with a test-signal (low-intensity and low-frequency sinusoids with electrode– specific frequencies) and a single sentinel electrode (not used for DC). Methods To validate this methodology, we developed lumped-parameter models of two and multi-electrode tDCS. Approaches with and without a sentinel electrode were solved and underlying assumptions identified. Assumptions were tested and parameterized in healthy participants using forearm stimulation combining tDCS (2 mA) and sinusoidal test-signals (38 μA and 76 μA peak to peak at 1 Hz, 10 Hz, and 100 Hz) and an in vitro test (where varied electrode failure modes were created). DC and AC component voltages across the electrodes were compared and participants were asked to rate subjective pain. Results A sentinel electrode is required to isolate electrode resistance in a two-electrode tDCS system. For multi-electrode resistance tracking, cross talk was aggravated with electrode proximity and current/resistance mismatches, but could be corrected using proposed approaches. Average voltage and average pain scores were not significantly different across test current intensities and frequencies (two-way repeated measures ANOVA) indicating the

  13. Pip, a novel IRF family member, is a lymphoid-specific, PU.1-dependent transcriptional activator.

    PubMed

    Eisenbeis, C F; Singh, H; Storb, U

    1995-06-01

    The immunoglobulin light-chain gene enhancers E kappa 3', E lambda 2-4, and E lambda 3-1 contain a conserved cell type-specific composite element essential for their activities. This element binds a B cell-specific heterodimeric protein complex that consists of the Ets family member PU.1 and a second factor (NF-EM5), whose participation in the formation of the complex is dependent on the presence of DNA-bound PU.1. In this report we describe the cloning and characterization of Pip (PU.1 interaction partner), a lymphoid-specific protein that is most likely NF-EM5. As expected, the Pip protein binds the composite element only in the presence of PU.1; furthermore, the formation of this ternary complex is critically dependent on phosphorylation of PU.1 at serine-148. The Pip gene is expressed specifically in lymphoid tissues in both B- and T-cell lines. When coexpressed in NIH-3T3 cells, Pip and PU.1 function as mutually dependent transcription activators of the composite element. The amino-terminal DNA-binding domain of Pip exhibits a high degree of homology to the DNA-binding domains of members of the interferon regulatory factor (IRF) family, which includes IRF-1, IRF-2, ICSBP, and ISGF3 gamma. PMID:7797077

  14. Rapid, Specific, No-wash, Far-red Fluorogen Activation in Subcellular Compartments by Targeted Fluorogen Activating Proteins

    PubMed Central

    2015-01-01

    Live cell imaging requires bright photostable dyes that can target intracellular organelles and proteins with high specificity in a no-wash protocol. Organic dyes possess the desired photochemical properties and can be covalently linked to various protein tags. The currently available fluorogenic dyes are in the green/yellow range where there is high cellular autofluorescence and the near-infrared (NIR) dyes need to be washed out. Protein-mediated activation of far-red fluorogenic dyes has the potential to address these challenges because the cell-permeant dye is small and nonfluorescent until bound to its activating protein, and this binding is rapid. In this study, three single chain variable fragment (scFv)-derived fluorogen activating proteins (FAPs), which activate far-red emitting fluorogens, were evaluated for targeting, brightness, and photostability in the cytosol, nucleus, mitochondria, peroxisomes, and endoplasmic reticulum with a cell-permeant malachite green analog in cultured mammalian cells. Efficient labeling was achieved within 20–30 min for each protein upon the addition of nM concentrations of dye, producing a signal that colocalized significantly with a linked mCerulean3 (mCer3) fluorescent protein and organelle specific dyes but showed divergent photostability and brightness properties dependent on the FAP. These FAPs and the ester of malachite green dye (MGe) can be used as specific, rapid, and wash-free labels for intracellular sites in live cells with far-red excitation and emission properties, useful in a variety of multicolor experiments. PMID:25650487

  15. A direct measurement of strontium-89 activity in bone metastases.

    PubMed

    Ben-Josef, E; Maughan, R L; Vasan, S; Porter, A T

    1995-06-01

    The total absorbed dose after systemic administration of 89Sr has been determined by measuring directly its activity in bone metastases. Autoradiography was performed on sections of bones obtained from patients treated with 89Sr to study the pattern of deposition. Discs of 5 and 8 mm diameter were cut from metastatic sites and normal bone. The beta-ray activity was determined with a scintillation counter, which was calibrated using similar bovine cancellous bone discs, onto which a known activity of 89Sr was transferred by pipette. From the activity measured, the initial activity (at the time of 89Sr administration) was calculated. The absorbed dose was estimated using the methodology described in NCRP Report No. 58. The estimated initial activity of 89Sr in the bone metastases varied from 2.3 to 240 MBq kg-1, with a mean value of 31 +/- 27 MBq kg-1. The total absorbed dose ranged from 1.3 to 64 Gy, with a mean of 18 +/- 16 Gy. The average total dose to normal bone sites was 1.1 +/- 0.4 Gy. The metastases to normal bone dose ratio in individual samples varied from 8 +/- 4 to 40 +/- 25. These estimates are in agreement with those obtained previously by indirect methods. PMID:7675358

  16. Cloning and expression of hybrid streptokinase towards clot-specific activity.

    PubMed

    Buniya, Harith K; Murugan, Vadivel; Thangadurai, Chinnathambi

    2014-03-01

    Streptokinase (SK) is a thrombolytic agent that is widely used to treat myocardial infarction and pulmonary embolism. The lack of fibrin specificity of SK for the clot lysis is one of the limitations of SK. In this study, we have incorporated the finger and Kringle 2 domains from the human tissue type plasminogen activator gene (t-PA) at the 5' end of the SK gene. These domains are responsible for specific binding to fibrin. We have used the pRSETB vector in an attempt to express the hybrid streptokinase possessing specificity for fibrin. On this regard, three hybrid streptokinase were constructed and expressed in Escherichia coli BL21 (DE3): the finger domain with SK (FSK), the Kringle 2 domain with SK (KSK) and the finger domain+Kringle 2 with SK (FKSK). The activities of the hybrid SKs were assessed by caseinolytic assay and clot lysis assay. All hybrid SKs were found to activate plasminogen in the caseinolytic plate assay. In the clot lysis assay, KSK and FSK were able to dissolute human blood and artificial clots in a fibrin-dependent manner unlike the SK and FKSK proteins. PMID:24440164

  17. Sequestration of nanoparticles by an EPS matrix reduces the particle-specific bactericidal activity

    PubMed Central

    Wang, Qian; Kang, Fuxing; Gao, Yanzheng; Mao, Xuewei; Hu, Xiaojie

    2016-01-01

    Most artificial nanomaterials are known to exhibit broad-spectrum bactericidal activity; however, the defence mechanisms that bacteria use based on extracellular polymeric substances (EPS) to detoxify nanoparticles (NPs) are not well known. We ruled out the possibility of ion-specific bactericidal activity by showing the lack of equivalent dissolved zinc and silicon toxicity and determined the particle-specific toxicity of ZnO and SiO2 nanoparticles (ZnONPs/SiO2NPs) through dialysis isolation experiments. Surprisingly, the manipulation of the E. coli EPS (i.e., no EPS manipulation or EPS removal by sonication/centrifugation) showed that their particle-specific bactericidal activity could be antagonized by NP-EPS sequestration. The survival rates of pristine E. coli (no EPS manipulation) reached 65% (ZnONPs, 500 mg L−1) and 79% (SiO2NPs, 500 mg L−1), whereas survival rates following EPS removal by sonication/centrifugation were 11% and 63%, respectively. Transmission electron microscopy (TEM) combined with fluorescence micro-titration analysis and Fourier-transform infrared spectroscopy (FTIR) showed that protein-like substances (N-H and C-N in amide II) and secondary carbonyl groups (C=O) in the carboxylic acids of EPS acted as important binding sites that were involved in NP sequestration. Accordingly, the amount and composition of EPS produced by bacteria have important implications for the bactericidal efficacy and potential environmental effects of NPs. PMID:26856606

  18. Music training leads to the development of timbre-specific gamma band activity.

    PubMed

    Shahin, Antoine J; Roberts, Larry E; Chau, Wilkin; Trainor, Laurel J; Miller, Lee M

    2008-05-15

    Oscillatory gamma band activity (GBA, 30-100 Hz) has been shown to correlate with perceptual and cognitive phenomena including feature binding, template matching, and learning and memory formation. We hypothesized that if GBA reflects highly learned perceptual template matching, we should observe its development in musicians specific to the timbre of their instrument of practice. EEG was recorded in adult professional violinists and amateur pianists as well as in 4- and 5-year-old children studying piano in the Suzuki method before they commenced music lessons and 1 year later. The adult musicians showed robust enhancement of induced (non-time-locked) GBA, specifically to their instrument of practice, with the strongest effect in professional violinists. Consistent with this result, the children receiving piano lessons exhibited increased power of induced GBA for piano tones with 1 year of training, while children not taking lessons showed no effect. In comparison to induced GBA, evoked (time-locked) gamma band activity (30-90 Hz, approximately 80 ms latency) was present only in adult groups. Evoked GBA was more pronounced in musicians than non-musicians, with synchronization equally exhibited for violin and piano tones but enhanced for these tones compared to pure tones. Evoked gamma activity may index the physical properties of a sound and is modulated by acoustical training, while induced GBA may reflect higher perceptual learning and is shaped by specific auditory experiences. PMID:18375147

  19. Sequestration of nanoparticles by an EPS matrix reduces the particle-specific bactericidal activity.

    PubMed

    Wang, Qian; Kang, Fuxing; Gao, Yanzheng; Mao, Xuewei; Hu, Xiaojie

    2016-01-01

    Most artificial nanomaterials are known to exhibit broad-spectrum bactericidal activity; however, the defence mechanisms that bacteria use based on extracellular polymeric substances (EPS) to detoxify nanoparticles (NPs) are not well known. We ruled out the possibility of ion-specific bactericidal activity by showing the lack of equivalent dissolved zinc and silicon toxicity and determined the particle-specific toxicity of ZnO and SiO2 nanoparticles (ZnONPs/SiO2NPs) through dialysis isolation experiments. Surprisingly, the manipulation of the E. coli EPS (i.e., no EPS manipulation or EPS removal by sonication/centrifugation) showed that their particle-specific bactericidal activity could be antagonized by NP-EPS sequestration. The survival rates of pristine E. coli (no EPS manipulation) reached 65% (ZnONPs, 500 mg L(-1)) and 79% (SiO2NPs, 500 mg L(-1)), whereas survival rates following EPS removal by sonication/centrifugation were 11% and 63%, respectively. Transmission electron microscopy (TEM) combined with fluorescence micro-titration analysis and Fourier-transform infrared spectroscopy (FTIR) showed that protein-like substances (N-H and C-N in amide II) and secondary carbonyl groups (C=O) in the carboxylic acids of EPS acted as important binding sites that were involved in NP sequestration. Accordingly, the amount and composition of EPS produced by bacteria have important implications for the bactericidal efficacy and potential environmental effects of NPs. PMID:26856606

  20. Music training leads to the development of timbre-specific gamma band activity

    PubMed Central

    Shahin, Antoine J.; Roberts, Larry E.; Chau, Wilkin; Trainor, Laurel J.; Miller, Lee M.

    2015-01-01

    Oscillatory gamma band activity (GBA, 30–100Hz) has been shown to correlate with perceptual and cognitive phenomena including feature binding, template matching, and learning and memory formation. We hypothesized that if GBA reflects highly learned perceptual template matching, we should observe its development in musicians specific to the timbre of their instrument of practice. EEG was recorded in adult professional violinists and amateur pianists as well as in four- and five-year-old children studying piano in the Suzuki method before they commenced music lessons and one year later. The adult musicians showed robust enhancement of induced (non-time-locked) GBA, specifically to their instrument of practice, with the strongest effect in professional violinists. Consistent with this result, the children receiving piano lessons exhibited increased power of induced GBA for piano tones with one year of training, while children not taking lessons showed no effect. In comparison to induced GBA, evoked (time-locked) gamma band activity (30–90 Hz, ~80 ms latency) was present only in adult groups. Evoked GBA was more pronounced in musicians than non-musicians, with synchronization equally exhibited for violin and piano tones but enhanced for these tones compared to pure tones. Evoked gamma activity may index the physical properties of a sound and is modulated by acoustical training, while induced GBA may reflect higher perceptual learning and is shaped by specific auditory experiences. PMID:18375147

  1. Critical Velocity Is Associated With Combat-Specific Performance Measures in a Special Forces Unit.

    PubMed

    Hoffman, Mattan W; Stout, Jeffrey R; Hoffman, Jay R; Landua, Geva; Fukuda, David H; Sharvit, Nurit; Moran, Daniel S; Carmon, Erez; Ostfeld, Ishay

    2016-02-01

    The purpose of this study was to examine the relationship between critical velocity (CV) and anaerobic distance capacity (ADC) to combat-specific tasks (CST) in a special forces (SFs) unit. Eighteen male soldiers (mean ± SD; age: 19.9 ± 0.8 years; height: 177.6 ± 6.6 cm; body mass: 74.1 ± 5.8 kg; body mass index [BMI]: 23.52 ± 1.63) from an SF unit of the Israel Defense Forces volunteered to complete a 3-minute all-out run along with CST (2.5-km run, 50-m casualty carry, and 30-m repeated sprints with "rush" shooting [RPTDS]). Estimates of CV and ADC from the 3-minute all-out run were determined from data downloaded from a global position system device worn by each soldier, with CV calculated as the average velocity of the final 30 seconds of the run and ADC as the velocity-time integral above CV. Critical velocity exhibited significant negative correlations with the 2.5-km run time (r = -0.62, p < 0.01) and RPTDS time (r = -0.71, p < 0.01). In addition, CV was positively correlated with the average velocity during the 2.5-km run (r = 0.64, p < 0.01). Stepwise regression identified CV as the most significant performance measure associated with the 2.5-km run time, whereas BMI and CV measures were significant predictors of RPTDS time (R(2) = 0.67, p ≤ 0.05). Using the 3-minute all-out run as a testing measurement in combat, personnel may offer a more efficient and simpler way in assessing both aerobic and anaerobic capabilities (CV and ADC) within a relatively large sample. PMID:26049790

  2. Development of a Competitive Cystatin C-Specific Bioassay Suitable for Repetitive Measurements

    PubMed Central

    Damm, Tatjana; Spiegel, Holger; Barth, Stefan; Fischer, Rainer; Naehring, Joerg

    2016-01-01

    Human cystatin C (hCC), a cysteine protease inhibitor, has been proposed as a diagnostic marker because its serum levels correlate with certain cardiovascular and kidney diseases. All current hCC assays are based on ex vivo detection. Here we describe the generation and evaluation of antibodies that allow the repetitive binding and release of hCC and hCC-fusion proteins, a prerequisite for long-term measurement, which is required for compatibility with implantable biochip devices and for the development of innovative antibody-based assays suitable for continuous in vivo and in vitro monitoring. Recombinant hCC and hCC-fusion proteins were produced in Escherichia coli and HEK293T cells and were used to generate antibodies by hybridoma technology. After screening by indirect and sandwich ELISAs, 12 monoclonal hybridoma cell lines producing hCC-specific monoclonal antibodies were identified. To determine their hCC association and dissociation properties, the antibodies were analysed by surface plasmon resonance spectroscopy, revealing three with the desired fast binding and moderate-to-fast release characteristics. The analysis of binding and dissociation in the presence of hCC and hCC-fusion proteins using fluorescence-based replacement assays showed that mAb CyDI-4 was the most suitable for further analysis. The results showed that repetitive replacement on mAb CyDI-4 was possible and that most of the change in signal intensity occurred after 20–30 min. Furthermore, the suitability of mAb CyDI-4 for serum hCC measurement was confirmed by a fluorescence-based replacement assay using serially-diluted reference serum from the Institute for Reference Materials and Measurements (ERM-DA471/IFCC). Our results suggest that the assay covers the physiological and pathological ranges of hCC. PMID:26799562

  3. The length of a lantibiotic hinge region has profound influence on antimicrobial activity and host specificity

    PubMed Central

    Zhou, Liang; van Heel, Auke J.; Kuipers, Oscar P.

    2015-01-01

    Lantibiotics are ribosomally synthesized (methyl)lanthionine containing peptides which can efficiently inhibit the growth of Gram-positive bacteria. As lantibiotics kill bacteria efficiently and resistance to them is difficult to be obtained, they have the potential to be used in many applications, e.g., in pharmaceutical industry or food industry. Nisin can inhibit the growth of Gram-positive bacteria by binding to lipid II and by making pores in their membrane. The C-terminal part of nisin is known to play an important role during translocation over the membrane and forming pore complexes. However, as the thickness of bacterial membranes varies between different species and environmental conditions, this property could have an influence on the pore forming activity of nisin. To investigate this, the so-called “hinge region” of nisin (residues NMK) was engineered to vary from one to six amino acid residues and specific activity against different indicators was compared. Antimicrobial activity in liquid culture assays showed that wild type nisin is most active, while truncation of the hinge region dramatically reduced the activity of the peptide. However, one or two amino acids extensions showed only slightly reduced activity against most indicator strains. Notably, some variants (+2, +1, −1, −2) exhibited higher antimicrobial activity than nisin in agar well diffusion assays against Lactococcus lactis MG1363, Listeria monocytogenes, Enterococcus faecalis VE14089, Bacillus sporothermodurans IC4 and Bacillus cereus 4153 at certain temperatures. PMID:25688235

  4. А new Gal/GalNAc-specific lectin from the mussel Mytilus trossulus: Structure, tissue specificity, antimicrobial and antifungal activity.

    PubMed

    Chikalovets, Irina V; Kovalchuk, Svetlana N; Litovchenko, Alina P; Molchanova, Valentina I; Pivkin, Mikhail V; Chernikov, Oleg V

    2016-03-01

    In the present study, a new Gal/GalNAc specific lectin from the mussel Mytilus trossulus (designated as MTL) was identified, and its expression levels, both in tissues and toward pathogen stimulation, were then characterized. The MTL primary structure was determined via cDNA sequencing. Deduced sequence of 150 amino acid residues showed 89% similarity to lectins from the mussels Crenomytilus grayanus and Mytilus galloprovincialis that were the first members of a new family of zoolectins. The results indicated that the MTL might be involved in immune response toward pathogen infection, and it might perform different recognition specificity toward bacteria or fungi. PMID:26802895

  5. Interpretation of the source-specific substantive control measures of the Minamata Convention on Mercury.

    PubMed

    You, Mingqing

    2015-02-01

    Being persistent, toxic, and bio-accumulative, Mercury (Hg) seriously affects the environment and human health. Due to Hg's attribute of long-range environmental transport across national borders, especially through atmospheric transport, no country can fully protect its environment and human health with its own efforts, without global cooperation. The Minamata Convention on Mercury, which was formally adopted and opened for signature in October 2013, is the only global environmental regime on the control of Hg pollution. Its main substantive control measures are source-specific: its phasing-out, phasing-down, and other main substantive requirements all direct to specific categories of pollution sources through the regulation of specific sectors of the economy and social life. This Convention does not take a national quota approach to quantify the Parties' nationwide total allowable consumption or discharge of Hg or Hg compounds, nor does it quantify their nationwide total reduction requirements. This paper attempts to find the underlying reasons for this source-specific approach and offers two interpretations. One possible interpretation is that Hg might be a non-threshold pollutant, i.e., a pollutant without a risk-free value of concentration. The existence of a reference dose (RfD), reference concentration (RfC), provisional tolerable weekly intake (PTWI), minimal risk level (MRL) or other similar reference values of Hg does not necessarily mean that Hg cannot be regarded as non-threshold because such reference values have scientific uncertainties and may also involve policy considerations. Another interpretation is that Hg lacks a feasibly determinable total allowable quantity. There is evidence that negotiators might have treated Hg as non-threshold, or at least accepted that Hg lacks a feasibly determinable total allowable quantity: (1) The negotiators were informed about the serious situations of the current emissions, releases, and legacy deposition; (2

  6. Neuronal activity is required for the development of specific cortical interneuron subtypes.

    PubMed

    De Marco García, Natalia V; Karayannis, Theofanis; Fishell, Gord

    2011-04-21

    Electrical activity has been shown to regulate development in a variety of species and in various structures, including the retina, spinal cord and cortex. Within the mammalian cortex specifically, the development of dendrites and commissural axons in pyramidal cells is activity-dependent. However, little is known about the developmental role of activity in the other major cortical population of neurons, the GABA-producing interneurons. These neurons are morphologically and functionally heterogeneous and efforts over the past decade have focused on determining the mechanisms that contribute to this diversity. It was recently discovered that 30% of all cortical interneurons arise from a relatively novel source within the ventral telencephalon, the caudal ganglionic eminence (CGE). Owing to their late birth date, these interneurons populate the cortex only after the majority of other interneurons and pyramidal cells are already in place and have started to functionally integrate. Here we demonstrate in mice that for CGE-derived reelin (Re)-positive and calretinin (Cr)-positive (but not vasoactive intestinal peptide (VIP)-positive) interneurons, activity is essential before postnatal day 3 for correct migration, and that after postnatal day 3, glutamate-mediated activity controls the development of their axons and dendrites. Furthermore, we show that the engulfment and cell motility 1 gene (Elmo1), a target of the transcription factor distal-less homeobox 1 (Dlx1), is selectively expressed in Re(+) and Cr(+) interneurons and is both necessary and sufficient for activity-dependent interneuron migration. Our findings reveal a selective requirement for activity in shaping the cortical integration of specific neuronal subtypes. PMID:21460837

  7. Engineering neprilysin activity and specificity to create a novel therapeutic for Alzheimer's disease.

    PubMed

    Webster, Carl I; Burrell, Matthew; Olsson, Lise-Lotte; Fowler, Susan B; Digby, Sarah; Sandercock, Alan; Snijder, Arjan; Tebbe, Jan; Haupts, Ulrich; Grudzinska, Joanna; Jermutus, Lutz; Andersson, Christin

    2014-01-01

    Neprilysin is a transmembrane zinc metallopeptidase that degrades a wide range of peptide substrates. It has received attention as a potential therapy for Alzheimer's disease due to its ability to degrade the peptide amyloid beta. However, its broad range of peptide substrates has the potential to limit its therapeutic use due to degradation of additional peptides substrates that tightly regulate many physiological processes. We sought to generate a soluble version of the ectodomain of neprilysin with improved activity and specificity towards amyloid beta as a potential therapeutic for Alzheimer's disease. Extensive amino acid substitutions were performed at positions surrounding the active site and inner surface of the enzyme and variants screened for activity on amyloid beta 1-40, 1-42 and a variety of other physiologically relevant peptides. We identified several mutations that modulated and improved both enzyme selectivity and intrinsic activity. Neprilysin variant G399V/G714K displayed an approximately 20-fold improved activity on amyloid beta 1-40 and up to a 3,200-fold reduction in activity on other peptides. Along with the altered peptide substrate specificity, the mutant enzyme produced a markedly altered series of amyloid beta cleavage products compared to the wild-type enzyme. Crystallisation of the mutant enzyme revealed that the amino acid substitutions result in alteration of the shape and size of the pocket containing the active site compared to the wild-type enzyme. The mutant enzyme offers the potential for the more efficient degradation of amyloid beta in vivo as a therapeutic for the treatment of Alzheimer's disease. PMID:25089527

  8. Autocatalytic activity and substrate specificity of the pestivirus N-terminal protease N{sup pro}

    SciTech Connect

    Gottipati, Keerthi; Acholi, Sudheer; Ruggli, Nicolas; Choi, Kyung H.

    2014-03-15

    Pestivirus N{sup pro} is the first protein translated in the viral polypeptide, and cleaves itself off co-translationally generating the N-terminus of the core protein. Once released, N{sup pro} blocks the host's interferon response by inducing degradation of interferon regulatory factor-3. N{sup pro'}s intracellular autocatalytic activity and lack of trans-activity have hampered in vitro cleavage studies to establish its substrate specificity and the roles of individual residues. We constructed N{sup pro}-GFP fusion proteins that carry the authentic cleavage site and determined the autoproteolytic activities of N{sup pro} proteins containing substitutions at the predicted catalytic sites Glu22 and Cys69, at Arg100 that forms a salt bridge with Glu22, and at the cleavage site Cys168. Contrary to previous reports, we show that N{sup pro'}s catalytic activity does not involve Glu22, which may instead be involved in protein stability. Furthermore, N{sup pro} does not have specificity for Cys168 at the cleavage site even though this residue is conserved throughout the pestivirus genus. - Highlights: • N{sup pro'}s autoproteolysis is studied using N{sup pro}-GFP fusion proteins. • N-terminal 17 amino acids are dispensable without loss of protease activity. • The putative catalytic residue Glu22 is not involved in protease catalysis. • No specificity for Cys168 at the cleavage site despite evolutionary conservation. • N{sup pro} prefers small amino acids with non-branched beta carbons at the P1 position.

  9. Organ-specific adaptive signaling pathway activation in metastatic breast cancer cells

    PubMed Central

    Burnett, Riesa M.; Craven, Kelly E.; Krishnamurthy, Purna; Goswami, Chirayu P.; Badve, Sunil; Crooks, Peter; Mathews, William P.; Bhat-Nakshatri, Poornima; Nakshatri, Harikrishna

    2015-01-01

    Breast cancer metastasizes to bone, visceral organs, and/or brain depending on the subtype, which may involve activation of a host organ-specific signaling network in metastatic cells. To test this possibility, we determined gene expression patterns in MDA-MB-231 cells and its mammary fat pad tumor (TMD-231), lung-metastasis (LMD-231), bone-metastasis (BMD-231), adrenal-metastasis (ADMD-231) and brain-metastasis (231-BR) variants. When gene expression between metastases was compared, 231-BR cells showed the highest gene expression difference followed by ADMD-231, LMD-231, and BMD-231 cells. Neuronal transmembrane proteins SLITRK2, TMEM47, and LYPD1 were specifically overexpressed in 231-BR cells. Pathway-analyses revealed activation of signaling networks that would enable cancer cells to adapt to organs of metastasis such as drug detoxification/oxidative stress response/semaphorin neuronal pathway in 231-BR, Notch/orphan nuclear receptor signals involved in steroidogenesis in ADMD-231, acute phase response in LMD-231, and cytokine/hematopoietic stem cell signaling in BMD-231 cells. Only NF-κB signaling pathway activation was common to all except BMD-231 cells. We confirmed NF-κB activation in 231-BR and in a brain metastatic variant of 4T1 cells (4T1-BR). Dimethylaminoparthenolide inhibited NF-κB activity, LYPD1 expression, and proliferation of 231-BR and 4T1-BR cells. Thus, transcriptome change enabling adaptation to host organs is likely one of the mechanisms associated with organ-specific metastasis and could potentially be targeted therapeutically. PMID:25926557

  10. Organ-specific adaptive signaling pathway activation in metastatic breast cancer cells.

    PubMed

    Burnett, Riesa M; Craven, Kelly E; Krishnamurthy, Purna; Goswami, Chirayu P; Badve, Sunil; Crooks, Peter; Mathews, William P; Bhat-Nakshatri, Poornima; Nakshatri, Harikrishna

    2015-05-20

    Breast cancer metastasizes to bone, visceral organs, and/or brain depending on the subtype, which may involve activation of a host organ-specific signaling network in metastatic cells. To test this possibility, we determined gene expression patterns in MDA-MB-231 cells and its mammary fat pad tumor (TMD-231), lung-metastasis (LMD-231), bone-metastasis (BMD-231), adrenal-metastasis (ADMD-231) and brain-metastasis (231-BR) variants. When gene expression between metastases was compared, 231-BR cells showed the highest gene expression difference followed by ADMD-231, LMD-231, and BMD-231 cells. Neuronal transmembrane proteins SLITRK2, TMEM47, and LYPD1 were specifically overexpressed in 231-BR cells. Pathway-analyses revealed activation of signaling networks that would enable cancer cells to adapt to organs of metastasis such as drug detoxification/oxidative stress response/semaphorin neuronal pathway in 231-BR, Notch/orphan nuclear receptor signals involved in steroidogenesis in ADMD-231, acute phase response in LMD-231, and cytokine/hematopoietic stem cell signaling in BMD-231 cells. Only NF-κB signaling pathway activation was common to all except BMD-231 cells. We confirmed NF-κB activation in 231-BR and in a brain metastatic variant of 4T1 cells (4T1-BR). Dimethylaminoparthenolide inhibited NF-κB activity, LYPD1 expression, and proliferation of 231-BR and 4T1-BR cells. Thus, transcriptome change enabling adaptation to host organs is likely one of the mechanisms associated with organ-specific metastasis and could potentially be targeted therapeutically. PMID:25926557

  11. Crosstalk among electrical activity, trophic factors and morphogenetic proteins in the regulation of neurotransmitter phenotype specification.

    PubMed

    Borodinsky, Laura N; Belgacem, Yesser H

    2016-04-01

    Morphogenetic proteins are responsible for patterning the embryonic nervous system by enabling cell proliferation that will populate all the neural structures and by specifying neural progenitors that imprint different identities in differentiating neurons. The adoption of specific neurotransmitter phenotypes is crucial for the progression of neuronal differentiation, enabling neurons to connect with each other and with target tissues. Preliminary neurotransmitter specification originates from morphogen-driven neural progenitor specification through the combinatorial expression of transcription factors according to morphogen concentration gradients, which progressively restrict the identity that born neurons adopt. However, neurotransmitter phenotype is not immutable, instead trophic factors released from target tissues and environmental stimuli change expression of neurotransmitter-synthesizing enzymes and specific vesicular transporters modifying neuronal neurotransmitter identity. Here we review studies identifying the mechanisms of catecholaminergic, GABAergic, glutamatergic, cholinergic and serotonergic early specification and of the plasticity of these neurotransmitter phenotypes during development and in the adult nervous system. The emergence of spontaneous electrical activity in developing neurons recruits morphogenetic proteins in the process of neurotransmitter phenotype plasticity, which ultimately equips the nervous system and the whole organism with adaptability for optimal performance in a changing environment. PMID:26686293

  12. Cellular Telephones Measure Activity and Lifespace in Community-Dwelling Adults: Proof of Principle

    PubMed Central

    Schenk, Ana Katrin; Witbrodt, Bradley C.; Hoarty, Carrie A.; Carlson, Richard H.; Goulding, Evan H.; Potter, Jane F.; Bonasera, Stephen J.

    2011-01-01

    OBJECTIVES To describe a system that uses off-the-shelf sensor and telecommunication technologies to continuously measure individual lifespace and activity levels in a novel way. DESIGN Proof of concept involving three field trials of 30, 30, and 21 days. SETTING Omaha, Nebraska, metropolitan and surrounding rural region. PARTICIPANTS Three participants (48-year-old man, 33-year-old woman, and 27-year-old male), none with any functional limitations. MEASUREMENTS Cellular telephones were used to detect in-home position and in-community location and to measure physical activity. Within the home, cellular telephones and Bluetooth transmitters (beacons) were used to locate participants at room-level resolution. Outside the home, the same cellular telephones and global positioning system (GPS) technology were used to locate participants at a community-level resolution. Physical activity was simultaneously measured using the cellular telephone accelerometer. RESULTS This approach had face validity to measure activity and lifespace. More importantly, this system could measure the spatial and temporal organization of these metrics. For example, an individual’s lifespace was automatically calculated across multiple time intervals. Behavioral time budgets showing how people allocate time to specific regions within the home were also automatically generated. CONCLUSION Mobile monitoring shows much promise as an easily deployed system to quantify activity and lifespace, important indicators of function, in community-dwelling adults. PMID:21288235

  13. Measurements of the PLT and PDX device activation

    SciTech Connect

    Stavely, J.; Barnes, C.W.; Chrien, R.E.; Strachan, J.D.

    1981-09-01

    Measurements of the activation levels around the PLT and PDX tokamaks have been made using a Ge(Li) gamma spectrometer and a Geiger counter. The activation results from radiation induced in the plasma by 14 MeV neutrons from the d(t,n)..cap alpha.. fusion reaction, 14.7 MeV protons from the d(/sup 3/He,p)..cap alpha.. fusion reaction, 10 ..-->.. 20 MeV hard x-rays from runaway electron induced bremmstrahlung, and 2.5 MeV neutrons from the d(d,n)/sup 3/He fusion reaction. The magnitude of the activation is compared to that predicted for PDX on the basis of one-dimensional activation codes.

  14. Comparing measured and modelled soil carbon: which site-specific variables are linked to high stability?

    NASA Astrophysics Data System (ADS)

    Robertson, Andy; Schipanski, Meagan; Ma, Liwang; Ahuja, Lajpat; McNamara, Niall; Smith, Pete; Davies, Christian

    2016-04-01

    Changes in soil carbon (C) stocks have been studied in depth over the last two decades, as net greenhouse gas (GHG) sinks are highlighted to be a partial solution to the causes of climate change. However, the stability of this soil C is often overlooked when measuring these changes. Ultimately a net sequestration in soils is far less beneficial if labile C is replacing more stable forms. To date there is no accepted framework for measuring soil C stability, and as a result there is considerable uncertainty associated with the simulated impacts of land management and land use change when using process-based systems models. However, a recent effort to equate measurable soil C fractions to model pools has generated data that help to assess the impacts of land management, and can ultimately help to reduce the uncertainty of model predictions. Our research compiles this existing fractionation data along with site metadata to create a simplistic statistical model able to quantify the relative importance of different site-specific conditions. Data was mined from 23 published studies and combined with original data to generate a dataset of 100+ land use change sites across Europe. For sites to be included they required soil C fractions isolated using the Zimmermann et al. (2007) method and specific site metadata (mean annual precipitation, MAP; mean annual temperature, MAT; soil pH; land use; altitude). Of the sites, 75% were used to develop a generalized linear mixed model (GLMM) to create coefficients where site parameters can be used to predict influence on the measured soil fraction C stocks. The remaining 25% of sites were used to evaluate uncertainty and validate this empirical model. Further, four of the aforementioned sites were used to simulate soil C dynamics using the RothC, DayCent and RZWQM2 models. A sensitivity analysis (4096 model runs for each variable applying Latin hypercube random sampling techniques) was then used to observe whether these models place

  15. Feature-specific imaging: Extensions to adaptive object recognition and active illumination based scene reconstruction

    NASA Astrophysics Data System (ADS)

    Baheti, Pawan K.

    Computational imaging (CI) systems are hybrid imagers in which the optical and post-processing sub-systems are jointly optimized to maximize the task-specific performance. In this dissertation we consider a form of CI system that measures the linear projections (i.e., features) of the scene optically, and it is commonly referred to as feature-specific imaging (FSI). Most of the previous work on FSI has been concerned with image reconstruction. Previous FSI techniques have also been non-adaptive and restricted to the use of ambient illumination. We consider two novel extensions of the FSI system in this work. We first present an adaptive feature-specific imaging (AFSI) system and consider its application to a face-recognition task. The proposed system makes use of previous measurements to adapt the projection basis at each step. We present both statistical and information-theoretic adaptation mechanisms for the AFSI system. The sequential hypothesis testing framework is used to determine the number of measurements required for achieving a specified misclassification probability. We demonstrate that AFSI system requires significantly fewer measurements than static-FSI (SFSI) and conventional imaging at low signal-to-noise ratio (SNR). We also show a trade-off, in terms of average detection time, between measurement SNR and adaptation advantage. Experimental results validating the AFSI system are presented. Next we present a FSI system based on the use of structured light. Feature measurements are obtained by projecting spatially structured illumination onto an object and collecting all of the reflected light onto a single photodetector. We refer to this system as feature-specific structured imaging (FSSI). Principal component features are used to define the illumination patterns. The optimal LMMSE operator is used to generate object estimates from the measurements. We demonstrate that this new imaging approach reduces imager complexity and provides improved image

  16. The exosome contains domains with specific endoribonuclease, exoribonuclease and cytoplasmic mRNA decay activities.

    PubMed

    Schaeffer, Daneen; Tsanova, Borislava; Barbas, Ana; Reis, Filipa Pereira; Dastidar, Eeshita Ghosh; Sanchez-Rotunno, Maya; Arraiano, Cecília Maria; van Hoof, Ambro

    2009-01-01

    The eukaryotic exosome is a ten-subunit 3' exoribonucleolytic complex responsible for many RNA-processing and RNA-degradation reactions. How the exosome accomplishes this is unknown. Rrp44 (also known as Dis3), a member of the RNase II family of enzymes, is the catalytic subunit of the exosome. We show that the PIN domain of Rrp44 has endoribonucleolytic activity. The PIN domain is preferentially active toward RNA with a 5' phosphate, suggesting coordination of 5' and 3' processing. We also show that the endonuclease activity is important in vivo. Furthermore, the essential exosome subunit Csl4 does not contain any domains that are required for viability, but its zinc-ribbon domain is required for exosome-mediated mRNA decay. These results suggest that specific exosome domains contribute to specific functions, and that different RNAs probably interact with the exosome differently. The combination of an endoRNase and an exoRNase activity seems to be a widespread feature of RNA-degrading machines. PMID:19060898

  17. Lysine-specific demethylase 1 promotes brown adipose tissue thermogenesis via repressing glucocorticoid activation.

    PubMed

    Zeng, Xing; Jedrychowski, Mark P; Chen, Yi; Serag, Sara; Lavery, Gareth G; Gygi, Steve P; Spiegelman, Bruce M

    2016-08-15

    Brown adipocytes display phenotypic plasticity, as they can switch between the active states of fatty acid oxidation and energy dissipation versus a more dormant state. Cold exposure or β-adrenergic stimulation favors the active thermogenic state, whereas sympathetic denervation or glucocorticoid administration promotes more lipid accumulation. Our understanding of the molecular mechanisms underlying these switches is incomplete. Here we found that LSD1 (lysine-specific demethylase 1), a histone demethylase, regulates brown adipocyte metabolism in two ways. On the one hand, LSD1 associates with PRDM16 to repress expression of white fat-selective genes. On the other hand, LSD1 represses HSD11B1 (hydroxysteroid 11-β-dehydrogenase isozyme 1), a key glucocorticoid-activating enzyme, independently from PRDM16. Adipose-specific ablation of LSD1 impaired mitochondrial fatty acid oxidation capacity of the brown adipose tissue, reduced whole-body energy expenditure, and increased fat deposition, which can be significantly alleviated by simultaneously deleting HSD11B1. These findings establish a novel regulatory pathway connecting histone modification and hormone activation with mitochondrial oxidative capacity and whole-body energy homeostasis. PMID:27566776

  18. Activity Level-Dependent Synapse-Specific AMPA Receptor Trafficking Regulates Transmission Kinetics

    PubMed Central

    Zhu, J. Julius

    2009-01-01

    Central glutamatergic synapses may express AMPA-sensitive glutamate receptors (AMPA-Rs) with distinct gating properties and exhibit different transmission dynamics, which are important for computing various synaptic inputs received at different populations of synapses. However, how glutamatergic synapses acquire AMPA-Rs with distinct kinetics to influence synaptic integration remains poorly understood. Here I report synapse-specific trafficking of distinct AMPA-Rs in rat cortical layer 4 stellate and layer 5 pyramidal neurons. The analysis indicates that in single layer 4 stellate neurons thalamocortical synapses generate faster synaptic responses than intracortical synapses. Moreover, GluR1-containing AMPA-Rs traffic selectively into intracortical synapses, and this process requires sensory experience-dependent activity and slows down transmission kinetics. GluR4-containing AMPA-Rs traffic more heavily into thalamocortical synapses than intracortical synapses, and this process requires spontaneous synaptic activity and speeds up transmission kinetics. GluR2-containing AMPA-Rs traffic equally into both thalamocortical and intracortical synapses, and this process requires no synaptic activity and resets transmission kinetics. Notably, synaptic trafficking of distinct AMPA-Rs differentially regulates synaptic integration. Thus, synapse-specific AMPA-R trafficking coarsely sets and synaptic activity finely tunes transmission kinetics and integration properties at different synapses in central neurons. PMID:19439609

  19. Measuring In Vitro ATPase Activity for Enzymatic Characterization.

    PubMed

    Rule, Chelsea S; Patrick, Marcella; Sandkvist, Maria

    2016-01-01

    Adenosine triphosphate-hydrolyzing enzymes, or ATPases, play a critical role in a diverse array of cellular functions. These dynamic proteins can generate energy for mechanical work, such as protein trafficking and degradation, solute transport, and cellular movements. The protocol described here is a basic assay for measuring the in vitro activity of purified ATPases for functional characterization. Proteins hydrolyze ATP in a reaction that results in inorganic phosphate release, and the amount of phosphate liberated is then quantitated using a colorimetric assay. This highly adaptable protocol can be adjusted to measure ATPase activity in kinetic or endpoint assays. A representative protocol is provided here based on the activity and requirements of EpsE, the AAA+ ATPase involved in Type II Secretion in the bacterium Vibrio cholerae. The amount of purified protein needed to measure activity, length of the assay and the timing and number of sampling intervals, buffer and salt composition, temperature, co-factors, stimulants (if any), etc. may vary from those described here, and thus some optimization may be necessary. This protocol provides a basic framework for characterizing ATPases and can be performed quickly and easily adjusted as necessary. PMID:27584824

  20. Identification of specific activities associated with fall-related injuries, active component, U.S. Army, 2011.

    PubMed

    Hauschild, Veronique D; Schuh, Anna; Taylor, Bonnie J; Canham-Chervak, Michelle; Jones, Bruce H

    2016-06-01

    Although falls continue to be a leading mechanism of serious injuries among military populations, interventions must target activities or hazards that can be controlled or managed. This project aimed to identify activities most frequently associated with Army soldier fall-related injuries to prioritize prevention strategies for this substantial health burden. Narrative data from Army safety, medical evacuation, and casualty reporting systems were reviewed to select incidents meeting inclusion criteria and assign established codes. Nondeployed (n=988) and deployed (n=254) injury rates were not statistically different (2.20 per 1,000 non-deployed person-years [p-yrs], 2.21 per 1,000 deployed p-yrs, respectively). More than 75% of injuries were temporarily disabling fractures, sprains, and strains, primarily to lower extremities. The most frequent activities associated with non-deployed fall injuries were sports (e.g., snowboarding and basketball; 22%), parachuting (20%), walking/marching (19%), and climbing (15%). Ice and snow were the leading hazard (43%). The most common associated activities among deployed soldiers were occupational tasks (53%), walking/patrolling (24%), climbing (23%), and sports (17%). Specific interventions that target the activities and hazards identified in this investigation are suggested as priorities to reduce Army fall-related injuries. PMID:27362343